Abstracts of Scientific Presentations 2019 AALAS National Meeting Denver, Colorado

Poster Sessions group each of NSG and NSG-SGM3 mice were microchipped using a physical restraint device, and 2 groups of NSG-SGM3 mice were P1 People Advocating Wellness and Support: Compassion Fatigue microchipped under isoflurane anesthesia. One anesthetic group and Satisfaction Team was microchipped with a surgical skin preparation and the other

was not. Mice were monitored for weight loss, signs of infection, A Schoell* and complications, such as inflammation, barbering, or microchip

loss. After 2-8 wk, mice were euthanized for gross pathology, Zoetis, Kalamazoo, MI subcutaneous culture, and histology. The restraint device method

was then similarly validated for groups (n=20) of nude (J:NU) and People working in laboratory animal research are exposed to a humanized (NSG and NSG-SGM3) mice. Pathological assessment variety of situations and experiences that, over time, can cause did not reveal a significant difference in inflammation (P > 0.999) chronic stress and emotional exhaustion. These situations and between groups, and all culture results were negative. Weights experiences include frequent euthanasia of animals, creation and postimplantation did not vary significantly between groups (P > maintenance of animal disease models, unexpected adverse events 0.06). The only noteworthy complication was occasional microchip involving animals, and many more. These experiences can negatively loss immediately following implantation using the physical restraint affect a person and lead to compassion fatigue. Compassion fatigue is device, with no persisting negative effects. We concluded that all a form of secondary traumatic stress that can be experienced by those of the assessed microchipping methods were acceptable for use on helping people or animals in distress. To help support our staff who immunocompromised mice. may experience varying degrees of compassion fatigue, we formed

an internal team dedicated to addressing compassion fatigue and P3 Dominance Status Has No Effect on Data Produced by SIV- improving compassion satisfaction. The name of the team is People infected Macaque (Macaca mulatta) Model and Remains Constant Advocating Wellness and Support (PAWS). The mission is to be a over Course of Infection cross-function team to raise awareness, educate, and stimulate open

conversation about compassion fatigue and compassion satisfaction A McNamara*, B Carlson, S Flemming, EK Hutchinson, M Li, S within our organization. The team will work to support and identify Queen, E Shirk, R Adams, J Clements, J Mankowski, KA Metcalf Pate needs within our organization that can positively influence and

support those people experiencing compassion fatigue. To help Molecular and Comparative Pathobiology, Johns Hopkins University, steer the direction of the team and identify needs of our people, Baltimore, MD we conducted a needs assessment in partnership with an external

consultant. Our team has used the needs assessment to launch Establishment of a dominance hierarchy is a natural consequence of initiatives that help focus on our core ACE principle: awareness, socially housing macaques. Though social housing has been shown communication, and education around compassion fatigue and to improve welfare, reduce stress, and improve research within compassion satisfaction. In the long term, we hope to positively the SIV-infected macaque model of HIV infection, it is unclear if support and improve the lives of our employees through PAWS. dominance status has an influence on SIV infection and subsequently

could introduce variability into the data produced by the model. P2 Assessment of Microchip Placement Methods in We sought to determine if dominance status within a pair has an Immunocompromised Mice effect on the immunological parameters of SIV infection, and if the

dominance status of a given macaque remains constant throughout A Hernandez*1, P Torres2, P Collier2, M Campagna1 the course of infection. We hypothesized that dominance status may

affect immunological parameters. In addition, we hypothesized that 1Comparative Medicine and Quality, The Jackson Laboratory, dominance would remain unchanged from preinoculation through Sacramento, CA; 2The Jackson Laboratory, Sacramento, CA acute infection and suppression. We determined dominance status

prior to inoculation for 12 socially housed Rhesus macaques (Macaca Microchipping has become an increasingly popular means of mulatta) using an ethogram to score incidents of dominant behavior identification in laboratory mice. Compared to other identification at feeding time. Following inoculation with SIV, we repeated methods, such as ear tags, microchips are less likely to cause dominance scoring during acute infection and following viral irritation, fall out, or be misread. Microchipped mice can easily be suppression. We determined plasma and CSF viral loads using qRT- scanned, and any associated data can be linked directly to their PCR, CD4 T cell counts by flow cytometry, and platelet counts via a microchip numbers. General anesthesia is often considered to be complete blood count throughout infection. All data was normalized the most controlled method of restraint for microchip implantation, to each individual macaque’s baseline data and analyzed. There was but other options exist. In our facility, physical restraint devices no significant difference between dominant and subordinate animals are commonly used without complication in immunocompetent in plasma and CSF viral loads, CD4 T cell counts, or platelet counts. mice. However, special considerations are often needed when Furthermore, dominance status remained unchanged throughout translating methods to immunocompromised animals, such as usage SIV infection for all pairs. These results demonstrate that dominance of more stringent practices. Therefore, a study was conducted to status does not interfere with or alter the course of infection and assess complication rates in immunocompromised mice implanted that dominance status remains constant throughout infection. Thus, with microchips using either physical or chemical restraint, and dominance hierarchy is unlikely to affect data produced by the SIV- with or without surgical skin preparation. We hypothesized that infected macaque model, and concerns about increasing variability complication rates would be similar between these different of the data should not deter researchers from socially housing techniques. NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) and NSG-SGM3 macaques within the model. [NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/

MloySzJ] mice were divided into 4 groups (n=20 per group). One

607 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P4 Optimization of Intrathecal Administration in Rats Using approach for “sticky” tracers that have a higher residual when other Fluoroscopic Guidance tubing or needle and syringe set ups are used; and a butterfly with saline and tongue depressor approach for obtaining and maintaining A Jones*, G Sweet, R Pielemeier venous access for anesthetized animals. When comparing these to traditional injection techniques, we have found less perivascular Surgery and Efficacy, Charles River Labs, Kalamazoo, MI infiltration, better image quality, less loss of tracers and radioactivity, and higher levels of proficiency by technicians Neurodegenerative diseases affect millions of people in the U.S. and the incidence continues to increase. Intrathecal injection (the P6 Optimizing Success in Nonhuman Primate Work with intended route in humans) can be used to target the central nervous Individualized Problem-solving Strategies system (CNS), providing analgesia, chemotherapy treatment, as well as gene and antibiotic therapy. Intrathecal administration allows AR Knight*, K Phataraphruk for smaller doses, avoids the need to cross the blood-brain barrier and may result in fewer systemic side effects. Potential efficacy School of Biological and Health Systems Engineering, Arizona State and safety of intrathecal administration test articles are frequently University, Tempe, AZ tested in rats, most typically done by unguided injection, surgical procedure, catheter, or port. Our facility has developed a method of Current training and restraint protocols for rhesus macaques (Macaca direct percutaneous intrathecal injection using fluoroscopic guidance. mulatta) typically involve generic solutions to behavioral challenges. To perform this method, the rats are anesthetized to effect with This 1-size-fits-all approach, while largely effective, still leaves room isoflurane. The animals are then placed in a ventral recumbency. for improvement. Working with nonhuman primates presents a A 27 g Lasse needle and a 100 µL gas tight syringe are filled with plethora of challenges that may benefit from a more individualized the test material. Using aseptic technique, the lumbar cistern is approach, particularly when an animal exhibits unique behaviors or accessed percutaneously under fluoroscopic guidance. Once the learning capabilities. Modifications to our current training and needle is in place, the test material is injected by slow bolus. The restraint protocols were inspired by 1 rhesus macaque that exhibited needle is then removed, and the animal recovered. To determine behavior characteristics that were counterintuitive to established proficiency during training, 100% of designated nonrecovery animals protocols. This prompted a customized training environment with an were successfully dosed with a 1:1 ratio of contrast and saline as emphasis on improving comfort, thereby minimizing distractions seen on the fluoroscopic images. Subsequently, a 100% success and enhancing focus. We hypothesized that these changes would rate was determined for designated recovery animals with no result in a more relaxed primate, allowing the animal to learn tasks lasting complications. Acute, recoverable complications, including more efficiently. Arm restraint tubes and full jackets are commonly temporary limb impairment and behavioral issues (isolation, shaking used tools for primate research. We found that modifying each of head, squinting eyes) were observed and were believed to be linked these items to be as minimally invasive as possible both increased the to increased fluid pressure within the intrathecal space due to the primate’s successful trials and decreased the amount of time spent on dose volume or secondary to needle insertion. Limiting dose volumes distraction-related behaviors. The promising results that we observed to 50 µL helps to minimize these effects. Our method of intrathecal with this new individualized approach have encouraged us to administration with fluoroscopic guidance increases the accuracy of develop more customized training procedures for each animal the dose, which reduces the number of animals required for a given moving forward. study, while also reducing stress to the animal. Since implementing this method, over 2,700 animals have been successfully dosed with P7 Making Compliance Forms More Compliance Friendly minimal complications lasting less than 24 h. Due to our success with rats, the procedure is being developed in mice. A Watts*, E Dohm, P Bhagat

P5 Specialized Injection Techniques for Preclinical Imaging University of Alabama at Birmingham, Birmingham, AL

AA Bedwell*, KM Eldridge, R Speedy, P Territo A common compliance issue reported by or to the major oversight agencies is deviation from the approved animal use protocol. While Indiana University School of Medicine, Indianapolis, IN this occurrence is often accidental divergence from the protocol by research staff, it can also be a result of the protocol form questions Preclinical imaging requires specific skills, knowledge, and and design. The electronic forms developed for compliance equipment in order to run effective and efficient studies. Appropriate registration and reporting for the IACUC have been simplified and technical skills are one of the most important aspects in collecting streamlined using conditional questions, guided responses, and high-quality data for interpretation and publication. Because many established policies, procedures, and program standards. Using these imaging probes, contrasts, and tracers are given IV in rodents, it is strategies to optimize the information collected for both research and important to have technicians who are proficient and accomplished auditing purposes gives researchers greater flexibility in performing at obtaining and maintaining venous access. In addition, specialized common, noninvasive procedures, reduces opportunities for techniques help ensure clean delivery of agents, with these deviation, and provides a more standardized data set that can be techniques being adjusted based on properties of the agents. easily queried and compared. Implementing this approach has Primarily, we use IV injections for position emission tomography helped reduce burden on the researchers while still maintaining a (PET) imaging, dynamic contract enhanced computerized high level of compliance, and has improved the committee’s ability tomography (DCE-CT), dynamic contract enhanced magnetic to assess animal welfare and perform program oversight. resonance imaging (DCE-MRI), and fluorescence imaging. Some PET and fluorescence tracers which are trapped or have slow clearance P8 A Refined Scalp Retractor for Mice can be delivered in conscious animals, while nontrappable PET tracers and probes for DCE-MRI and DCE-CT have to be delivered A Lapierre* on scanner in anesthetized animal. This means the additional challenge of maintaining venous access on an anesthetized animal. Surgical Services, The Jackson Laboratory, Bar Harbor, ME We have created specialized techniques that help overcome these obstacles, as well as agent dependent issues (viscosity, radioactive Mice are used extensively for neuroscience research and therefore, materials, sticking, residual, etc.). The techniques include a butterfly stereotaxic surgery is frequently employed. Current techniques catheter with saline approach for standard PET tracers and imaging generally call for retraction of the scalp with bulldog or similar style probes that ensure accuracy and prevent undue handling and clamps which results in trauma to the skin. Colibri style retractors are exposer to radioactive materials; an insulin needle direct stick atraumatic but the design is not optimal. The refinement of a scalp

608 Abstracts of scientific papers 2019 AALAS National Meeting

retractor for stereotaxic surgery has been an ongoing effort at our training records have increased by 25%. Compliance has improved facility. We designed a scalp retractor that is easily positioned and with better tracking of the 150 new and revised procedures and atraumatically retracts the scalp with substantial exposure of the policies. The increase in accountability that the electronic training skull. The shape, size, and tension imparted by the retractor is well record website provides has decreased the deficiency findings at the matched to the size of the mouse. Compared to Colibri-style IACUC’s semiannual inspections. During this transition, we’ve retractors and other methods of skin retraction, our design provides experienced training complexity within our department. For an easy to use, stable, atraumatic instrument for skull exposure. This example, differences in training needs within core groups improved retractor aligns with the 3Rs by minimizing tissue trauma (gnotobiotics, small animal imaging), the type of animal species while providing excellent exposure of the surgical site. (nonhuman primates, rodents, and frogs) and working with biohazards or hazardous chemical agents all contributed to this P9 Refined Surgical Approach for Combination Microdialysis/ complexity. The electronic training record website satisfied IACUC, Cisterna Magna Ported Nonhuman Primate Models AAALAC, OLAW, and Environment, Health and Safety requirement of employee training records in a central location. AN Bone* P11 Cannulock Catheter Buttons Improve Patency In Rats for SALAR, Merck, West Point, PA Longterm Infusion Dosing and Sampling Research Studies

A conscious nonhuman primate model allowing evaluations of B Gien* pharmaceutical compound levels in brain fluids can have tremendous impact in disease areas such as Parkinson’s, Alzheimer’s, Surgery, Envigo, Indianapolis, IN and other neurodegenerative disorders. Animals cannulated for microdialysis along with a cisterna magna port (CMP) can enable the The importance of the 3Rs in research is an ever-increasing goal of evaluation of neurochemicals, large peptides, and metabolites in the today’s global research community. According to the Foundation for interstitial fluid (ISF) compared to the cerebrospinal fluid (CSF) Biomedical Research (FBR), 95 percent of all lab animal used are which can provide a better understanding of pharmacokinetics and rodents, primarily mice and rats. Being able to extend the use of pharmacodynamics of compounds in the brain. Over the past 4 y we research models and reuse them when appropriate is key for developed a combined microdialysis and CMP rhesus macaque continuing to achieve the reduction and replacement of rodents in model by way of 2 different surgical preparations. The initial preclinical research. Using advanced technologies to create animals surgeries targeted the striatum. This is a small, deeper target region models that can improve animal welfare are also a key consideration so the approach involved the surgical placement of fiducial markers when planning and conducting research studies. Repeated sampling to facilitate the coordinate determination and design of a custom- and dosing in rodents, including longterm infusions, can be fitted recording chamber using MRI and CT scans, followed by a accomplished in a refined way by using newly refined catheters and second surgery for chamber placement, cannulation, and CMP. With exteriorization methods. Using cannulock buttons and medical-grade this approach, additional imaging is performed to evaluate the polyurethane catheters we extended catheter patency for longterm placement of the microdialysis cannula. The limiting factors studies and cross over studies. The cannulock button provided a associated with performing the microdialysis and CMP surgery in closed system that can be easily attached to any catheter and the same surgical session are the time required to customize the maintained a solid reliable connection because of the barbed stainless chamber, the extended surgery time and the inability to ensure steel connector.We used male Sprague-Dawley rats for this study functionality of the CMP resulting in surgical repairs. The next step with a 3 Fr medical grade polyurethane jugular vein catheter and a 3 was to evaluate if we need a customized head cap when targeting the Fr medical grade polyurethane femoral vein catheter. The cannulock cortex, a larger brain region. Elimination of the steps required for buttons were accessed twice weekly for 90 d postsurgery. On customization allowed us to modify the procedure to a 2-pronged accessing the catheters we used a locking solution consisting of surgical approach. First is the placement of the CMP. The animals heparin:glycerol and a flush solution of heparinized saline. Twice per recuperate and during this time we evaluate the patency of the week we removing the locking solution and flushed the catheters. catheter. If the CMP is functioning properly and the animal has After flushing the catheter we withdrew a small amount of blood to healed well, then we perform the microdialysis cannulation placing a complete a full patency check. The catheter was flushed with commercially available head cap. This head cap has a lower profile heparinized saline and relocked with the heparin glycerol solution. and the animals adapt quickly to it. The second approach allowed us At the end of 90 d all animals were euthanized and a necropsy was to reduce the amount of resources and time it took to complete the completed. The results confirmed that were able to extend patency surgeries and move the model to study. when compared to other methods of catheter exteriorization in rodents. We also noted improved animal welfare and technician P10 Bringing Employee Training Documentation into the Modern welfare due to the ease of use and less stressful method of accessing Age: Transitioning to an Electronic Training Record Website catheters.

BM Montague* P12 Divided Caging Improves Welfare in Male C57BL/6 Mice

LASP, NCI, Frederick, MD BR Tallent*1,2, L Law1,2, M Saber1, J Lifshitz1,2

A laboratory animal science program replaced the old method of 1Child Health, University of Arizona College of Medicine; 2Phoenix tracking employee training using paper-based documentation with Children’s Hospital, Phoenix, AZ an easily accessibe, flexible, and user-friendly electronic training record website. The previous paper-based training matrix used for Many devices have been introduced to elicit natural behaviors and new hires was outdated and inefficient. The goal was to create a reduce aggression to improve animal welfare, most with debatable program that would generate training requirements per job title, send results. We have previously demonstrated significant reductions in automated notifications of new and revised standard operating aggressive-like behaviors in group-housed male mice housed in procedures (SOP), and document hands-on technique training. A partially divided caging. Limitations of this pilot study included cage team was established with varying expertise; the team included data density, small population sizes, duration of study, and lack of management services, facility and program managers, veterinary behavioral and physiological data. To assess this, animals were raised staff, and the quality assurance office to help develop this electronic with either partial cage dividers or in standard housing with no training record website. The team implemented a workflow, tested divider. Following one week of acclimation in the vivarium, male data, and presented group demonstrations for functionality before mice were weaned at 21 d old into randomly assigned cages, either the launch in 2016. Since then, new employee electronic viewable standard cages or cages with a partial cage divider. Animal behavior

609 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 was tested on rotarod for balance and motor coordination, open field that are observed in people living with HIV. Macaques are a social and elevated plus maze for anxiety-related behavior, and novel object species, yet are often singly housed for infectious disease research. recognition and Y maze for cognitive behaviors. Body weights were Single housing is stressful for macaques, and stress has been shown taken from weaning to 90 d of age, and again from 133 to 180 d of to alter serotonin and dopamine metabolism. We have demonstrated age. Bite wounds were counted beginning at 133 d of age, as well as that singly housed macaques experience a more profound decline in video recordings assessing posturing, fighting, biting, and injury as the number of circulating CD4+ T cells, greater T cell activation, and aggressive behaviors. Results indicated significantly improved higher plasma and central nervous system (CNS) viral loads outcomes for anxiety-based testing, consistently higher body compared to socially housed macaques, and produce more variable weights, statistically fewer bite wounds, and a statistically significant data. We therefore hypothesized that social housing of SIV-infected decrease in identified aggressive behaviors from mice in partially macaques would cause changes in CNS dopamine and serotonin divided cages compared to mice in standard cages. We conclude that metabolism compared to single housing. We used high-performance reductions in anxiety-like behavior, aggressive behaviors, bite liquid chromatography (HPLC) to analyze banked cerebral spinal wounds, and increased body weights indicate an improvement in fluid samples for levels of dopamine, its precursor DOPA and its overall welfare of co-housed C57BL/6 male mice housed with partial metabolites homovanillic acid, norepinephrine and epinephrine, and cage dividers, when compared to mice housed in standard for essential components of the serotonin pathway, including its conditions. precursor tryptophan and tryptophan’s alternate metabolite, kynurenine. We detected increases in both kynurenine and its P13 Optimization of Intraarticular Administration Using precursor tryptophan on day 7 following inoculation with SIV in Fluoroscopic Guidance socially housed animals compared to singly housed animals (P < 0.0001), indicating that tryptophan is being shunted away from C DePlancke*, G Sweet, J Justen, S Woods serotonin and melatonin production in socially housed animals following SIV infection. This decrease in the proinflammatory Charles River Laboratories, Mattawan, MI metabolite kynurenine is consistent with the blunted immune response to SIV that we previously observed in singly housed Inflammatory joint conditions are common diseases that affect animals. These data indicate that both the immune response in the millions of people. Some of the most common diseases are CNS and the concentration of neurotransmitters are altered by social osteoarthritis, rheumatoid arthritis, and psoriatic arthritis. These housing, and thus that single housing of SIV-infected animals has the diseases are characterized by joint pain, stiffness, and swelling due to potential to affect the translational value of data produced by the damage of the joint. Intraarticular delivery is a route used to deliver model. potential new therapies for inflammatory joint conditions, which include synovial fluid replacement, stem cells, and viral vectors. The P15 Development of a Rabbit Enrichment Program and common method for this type of injection in a rat is to insert the Contribution to a Culture of Transparency and Care needle through the side of the knee joint, under the patellar ligament, into the joint space. However, this technique is difficult to master and T Worlds*2, F De La Garza2, K Kelly2, CZ Cannon1, M Jernigan2, G can be unreliable because it is easy to insert the needle beyond the Jernigan2 targeted joint space. We have found that inserting the needle through the patellar ligament provides an easier and more reliable method. To 1Becton Dickinson, Research Triangle Park, NC; 2Charles River accomplish this, the animal is anesthetized via Isoflurane inhalation Laboratories, Research Triangle Park, NC and placed in a dorsally recumbent position. The dose site is shaved and scrubbed with chlorhexidine solution and the hind limb is bent Rabbit models are used in the biomedical industry for good to allow the indent on the knee at the end of the patella to be located. laboratory practice (GLP) safety and toxicity testing, antibody The needle is then gently inserted through the space below the production, and preclinical studies. Rabbits are a prey species and patella and the needle will drop into the joint with no resistance. We can become easily stressed; therefore, emphasis must be placed on used contrast agent and fluoroscopic images to train individuals on well-acclimated rabbits. Providing an environment that encourages this method. This allowed us to confirm proper administration and species-specific behavior leads to more healthy rabbits, successful determine a maximum dose volume. Using these tools, we found anesthesia events, and reuse of rabbits for training and retirement that up to 100 ul can be injected into rats larger than 225 g with no opportunities. Animal care staff were challenged to improve rabbit visible leakage or side effects. We determined that this method can be acclimation and enrichment using limited resources. The goal was to mastered in a shorter period of time and results in more accurate introduce individually housed rabbits to a positive experience from administration allowing less animals to be needed on study. the moment they arrived into the animal care program. Enhancements included daily animal caretaker contact, visualization P14 Serotonergic Metabolism Differs in Pigtailed Macaques That of other rabbits, larger caging to promote normal behaviors, reuse of Are Singly Housed Compared to Socially House during Acute empty swine runs for exercise, and reuse of feed bins for safe Simian Immunodeficiency Virus Infection transportation within the facility. Once rabbits transitioned onto study, more restrictions applied, however daily caretaker contact C Lyons1, S Guerrero-Martin1, S Aja4, K McGee1, L Rubin2, S Queen1, continued that included brushing and playgroups. Rabbits were then E Shirk1, M Li1, B Bullock1, D Graham3, C Zink1, L Gama1, R Adams1, eligible for transfer onto a training protocol or for adoption. The J Clements1, J Mankowski1, KA Metcalf Pate*1 success of acclimation and daily enrichment by a variety of caretakers and research staff has resulted in well-socialized, approachable, and 1Molecular and Comparative Pathobiology, Johns Hopkins more healthy rabbits in a dynamic environment. Rabbits are easy and University School of Medicine, Baltimore, MD; 2Neurology- safe to handle by PIs and animal care staff resulting in efficient Neuroimmunology and Neurological Infections, Johns Hopkins husbandry and decreased stress as a scientific variable. Less stress University School of Medicine, Baltimore, MD; 3Anesthesiology and has reduced premedication drug volumes and shorter anesthesia Critical Care Medicine, Johns Hopkins University School of recovery time. The program promotes a culture of care and Medicine, Baltimore, MD; 4Neuroscience, Johns Hopkins University transparency. Rabbits have responded positively to social enrichment School of Medicine, Baltimore, MD with animal care staff, novel exercise opportunities, and a variety of feed enrichment. Transparency has led to the onsite cafeteria staff Simian immunodeficiency virus (SIV)-infected macaques are an participation in the animal care program by reuse of leafy greens and essential animal model in HIV cure research. They are expected to produce for feed enrichment. The novel enrichment program has reliably model changes to the immune system, such as CD4+ T cell contributed to the 3Rs by refining environmental enrichment decline, and changes in the central nervous system, such as opportunities leading to a reduction in rabbit numbers and reuse of perturbations of the dopamine and serotonin metabolism pathways rabbits for training or retirement purposes.

610 Abstracts of scientific papers 2019 AALAS National Meeting

P16 Renovating a Training Program: Reframing Training from surgical outcome requires appropriate attention to aseptic and SOPs to Ranked Skills Using a Novel Skill Scorecard Tool surgical techniques, including anesthetic monitoring and analgesic administration. Pre-warming the recovery cages also aids in the C Weiner*, A Sanderson ability of the mice to heal more efficiently after surgery. Our CSF collection technique allows us to produce the first proof of concept Taconic Biosciences, Inc., Rensselaer, NY for biomarkers of intrathecal inflammation and neurodegeneration that may be ultimately translated to human patients with MS. Our As employee tolerance for training based only on standard operating serial CSF collection technique also decreases the need for serial procedures (SOPs) decreases, the role of the trainer and training sacrifice, resulting in the use of a minimal number of mice, therefore programs must evolve. Many training programs within the impacting the 3R’s of animal research. laboratory animal sciences are typically driven by SOPs, and the timeline expectations for an employee to progress through training P18 Evaluating Savings a Year after Establishing an Inhouse are often generalized and not data-driven. Also, many training Rodent Registry programs are institution-based, which limits the transferability of tools and programs between institutions and the field overall. This D Duvall*1, A Watts2 can make periods of high employee turnover and/or planning for low workload downturns or high-volume ramp ups exceedingly 1Animal Resources Program, University of Alabama at Birmingham, difficult to plan against without significant impacts to quality, cost, Birmingham, AL; 2IACUC, University of Alabama at Birmingham, and successful outcomes. As a result, understanding the employee Birmingham, AL training process and ensuring the skill competency is a valuable planning tool that positively impacts animal use programs. In June 2018, a university animal resources program (ARP) and Therefore, a comprehensive and standardized training program must Institutional Animal Care and Use Committee (IACUC) established include a holistic approach to engagement, assessment, and an inhouse rodent registry, allowing investigators the ability to search opportunity that goes beyond the SOP. We propose a novel system a secure web page for specific rodent lines before looking to outside that reframes the job description and employer expectations into a sources for their research needs. Initially, the registry consisted of ranked list of skills. The training curriculum is then based on a ~1,750 rodent listings, but has now grown to ~2,300 rodent listings, standardized definition of skill-based training. Further, we propose with mice making up ~93% of the list. This registry does not account the use of a training skill scorecard that serves as the basis for a skill- for every line on campus, but has been a powerful tool used by the based curriculum. The skill scorecard consists of five parts: 1) Setting investigator(s) looking for a model that may benefit their research. the expectation by defining the needed training 2) Provide the Specially built queries extract information from the ARP animal order practical and didactic resources 3) Proficiency and competency database and IACUC electronic protocol forms to identify potentially demonstrations through skills assessments 4) Reporting metrics 5) available rodent lines. The information within the registry is Opportunities for internal and external growth and development. searchable by species and strain/line nomenclature. Of the 38 Finally, the training efforts must include a reference to the reason the investigator requests, 3 resulted in known transfers between UAB institution is asking the employee to learn each skill, whether that be investigators, 4 identified lines that were their own, 1 was for animals the company vision or program’s mission statement. The poster will not listed in the registry, 1 investigator had to import a line listed due outline: 1) the details of each of the parts of the skill scorecard to it being unavailable by the lab on campus, and 29 were for other definition, 2) an example list of ranked skills for animal care and reasons such as animal availability, or no transfer performed. At corresponding scorecard, and 3) an example of tracking and present, this registry has saved at least 3 investigators from having to reporting against employees in training. We hypothesize that skills import animals from outside sources. While this number seems based training will generate a more flexible workforce long term. small, the savings included invaluable time needed for their research along with cost savings totaling over $5,000 related to importing, P17 Development of an Improved Technique for Serial Survival quarantine, and health assessment fees. Based on the early, but CSF Collection in Mice positive results of the current registry, it will continue to be maintained and monitored to provide investigators with a valuable DB Royce*, K DiSano, F Gilli, A Pachner and beneficial research aid at our institution.

Neurology, Dartmouth Hitchcock Medical Center, Lebanon, NH P19 Comparison of the Efficacy of 2 Types of Antibiotic Mixtures in Gut Flora Depletion in Mice Multiple sclerosis (MS) is an immune-mediated disease affecting the brain and spinal cord that is associated with chronic inflammation D Goulding*, P Myers, A Dickerson, M Comins, R Wiltshire, K Laber, leading to demyelination and neurodegeneration. Clinical TL Blankenship-Paris management of MS is challenging because of the clinical heterogeneity and the rate of disease progression. Thus, in order to Comparative Medicine Branch, National Institute of Environmental improve the care of patients with MS, biomarkers that can predict Health Sciences, Durham, NC disease onset, exacerbation, progression, as well as response to treatments are necessary. Despite the obvious utility of easily There has been increased interest over the last decade of the role of accessible blood or urine-based biomarkers, so far results from these the microbiome in health and disease. The use of germfree models fluid compartments have not been informative. In contrast, and depletion of the microbial flora using antimicrobials are 2 substantial progress has been made in cerebrospinal fluid (CSF) methods commonly employed in studying the microbiome in the biomarkers, a fluid that surrounds the brain and spinal cord, i.e. the laboratory mouse. Germfree mice are born, raised, and studied in epicenter for the disease. Since MS biomarkers have the opportunity isolators in the absence of any known microbes; however, the to be developed, tested, and validated in mouse models of MS, we equipment, supplies, and training required for the use of germfree have optimized a refined cisterna magna puncture technique for CSF mice can be quite costly and time consuming. The use of antibiotics collection in the Theiler’s Murine Encephalomyelitis Virus-Induced to decrease the microbial flora does not require special equipment, is Demyelinating Disease (TMEV-IDD) model of progressive MS. This applicable to any mouse strain, and is relatively inexpensive. sampling technique allows serial CSF samples to be obtained from However, mice do still have microbes as they are not in a germfree the same mouse at 2-mo intervals. Amounts vary depending on size, environment. One commonly used antibiotic cocktail regime uses hydration, and health of the mouse, with an average collection ampicillin, neomycin, metronidazole, and vancomycin in the volume of 6 μl (2 to 10 μl). Minimizing the tissue damage during each drinking water for 4 wk. We have found the palatability of this surgery is essential to aid in wound healing and to reduce bleeding mixture to be poor resulting in weight loss of the mice requiring which increases the difficulty of obtaining clear CSF. A successful removal from study. The addition of sucralose to the drinking water

611 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 and supplying wet food (mash) made with this drinking water has C57Bl/6 background with an intact LVS with a proven stud male, improved the outcome. However, the palatability still results in a cull mouse also on the C57Bl/6 background. The number of high number of mice requiring removal. This study evaluated a new pregnancies, dystocia, and torn and intact LVS after parturition were combination of antibiotics to reduce the gut microbiota and improve recorded. This data, in addition to the incidence of LVS and dystocia animal welfare. C57BL/6NCrl mice were placed on 1 of the following in the original breeding colony, was used to perform a cost analysis drinking water regimes: ampicillin/neomycin/metronidazole/ and to determine the potential costs to the researcher of breeding a vancomycin water (n=13), enrofloxacin/ampicillin water (n=11), or mouse with a LVS. Furthermore, we created an inanimate tool to standard reverse osmosis deionized water (RODI) (n=11). Mice were train research and veterinary staff on how to examine the mouse for weighed and water consumption measured. Feces were collected the presence of a LVS. The addition of the training tool has led to an before and after 8 d of antibiotic treatment. Quantitative real-time increased comfort level in checking for this abnormality and in turn, PCR (RT-qPCR) for 16s bacterial ribosome was done on each sample more animals are examined and excluded from breeding. Overall, and compared. We found the combination of enrofloxacin and this saves the researcher costs associated with per diem rates and the ampicillin resulted in improved palatability, improved elimination of loss of productive research time while simultaneously improving the gut flora, and reduction in the gut flora at a more rapid rate. animal welfare.

P20 Improving the Welfare of Hamsters during Euthanasia P22 The Move from Instructor-led to Computer-based Training: Things to Consider When Developing an eLearning Program DL Hickman* D Ehleiter*, M Eckstein, N Lipman Laboratory Animal Resource Center, Indiana University, Indianapolis, IN Center of Comparative Medicine and Pathology, Memorial Sloan Kettering Cancer Center, New York, NY The most recent version of the AVMA Guidelines on Euthanasia recommend methods of euthanasia for laboratory rodents. The data A properly constituted training program significantly improves supporting these recommendations is based on a body of work that animal welfare and regulatory compliance; however, the training focuses primarily on rats and mice, though the guidelines extend needs of the investigative and husbandry staff must be balanced these recommendations to “small rodents.” Because there are against a limited number of trainers and time-restricted schedules. To signficant welfare differences between rats and mice, it follows that meet this goal, our training program has evolved from a strictly additional study is needed for recommendations specific to other instructor-led, classroom-based program to a blended learning laboratory rodents. This study evaluated the welfare of 72 hamsters method. Comprehensive online training modules are used for the (6 males and 6 females per group) that were euthanized by carbon initial introduction of information while instructor-led follow-up dioxide (30%, 50%, and 70% volume per minute displacement rate of sessions provide a review that emphasizes key concepts. This method

100% CO2), isoflurane (3% or 5% inhaled in 0.5L/min O2), or aids the learners in the application of information that is targeted to pentobarbital (354mg, IP). Behaviors (rearing, freezing/walking their specific needs, projects, and procedures. To ensure the backward, jumping, defecation) were recorded for each treatment. instructor-led sessions are brief and maximally productive, the online Physiologic measures (corticosterone, complete blood count, serum modules must be effective at presenting the information in a manner biochemistry) were also collected. All methods of euthanasia resulted that is conducive to comprehension. This presentation will review in approximated loss of consciousness within 104 sec.. A significant educational principles as they relate to computer-based training and increase in the proportion of rears and jumps was recorded in the will provide the rationale used to determine the best options to meet hamsters euthanized with 3% isoflurane. Both isoflurane groups had the institution’s training goals. Additionally, questions regarding a significant increase in fecal boli produced. Hamsters euthanized when to use videography, preferences for captioned versus narrated with 5% isoflurane and pentobarbital had significant leukopenia. formats, and how to determine the optimal length of a presentation/ There were no significant differences in the prorortion of rearing training session will be addressed. behaviors or in corticosterone levels. Creatinine levels were P23 A Pilot Program to Certify Rodent Surgery Trainers significantly increased in the hamsters euthanized with 5% isoflurane. Phosphorous and calcium were significantly decreased in E Karlsson*, K Cataline, L Carbone the hamsters euthanized with 5% isoflurane. ALP was significantly decreased in the hamsters euthanized with pentobarbital. Overall, Institutional Animal Care and Use Program, University of California, this study suggests that all three methods of euthanasia are San Francisco, San Francisco, CA appropriate options for the euthanasia of hamsters, though there may be welfare concerns associated with the use of isoflurane, especially Even with a robust institutional surgery training program for animal at lower concentrations, that require future study. users, individual labs must still train their personnel to perform the labs’ specific procedures. This creates a culture of tribal knowledge P21 Longitudinal Vaginal Septa in Mice: Identifying the “Y” in the transfer, where surgery practices often do not adhere to standards Road Saves Time, Money, and Avoids Distress to the Animal known to produce the best outcomes. This situation persists when institutions lack the resources or know-how to provide lab-specific DR Totman*, EK Daugherity training to every to-be surgeon. To address this, we piloted a program to certify individuals within labs as surgery trainers, CARE, Cornell University, Ithaca, NY offering a resource-efficient method for proposing, implementing, and disseminating surgery method refinements. In the pilot program, Longitudinal vaginal septa (LVS) are bands of tissue of varying an IACUC veterinarian provided train the trainer instruction that thickness that bisect the vaginal canal longitudinally, creating a “Y” focused on aseptic technique, analgesia, and supportive care. One- in the vaginal canal. Histologically, LVS consist of fibrous connective on-one meetings allowed the veterinarian to assess surgical skills and tissue covered by vaginal mucosa. LVSs have been shown to increase protocol compliance, and provide feedback to improve welfare and the rate of dystocia in mice causing distress to the animal. Here, address the labs’ research goals. Candidate trainers were asked to breeding coordinators for hire by research staff routinely examine develop lab-specific checklists, training references, and standard females for a LVS before breeding and eliminate these mice from operating procedures. The process ended with the development of breeding colonies. This is done to avoid lost research time, procedure-specific proficiency benchmarks that are verified prior to unnecessary per diem charges, and to prevent health complications certifying individuals to perform surgery independently. The and pain to the female mouse due to the risk of dystocia. In order to program critically depends on enthusiastic lab participation to be determine if dystocia occurs in every female from a large breeding effective. Two factors we explored were 1) compliance history and 2) coordinator-maintained colony, we bred 9 female mice on the whether or not labs were required by the IACUC to participate. We

612 Abstracts of scientific papers 2019 AALAS National Meeting

first invited labs to volunteer with the option of not participating at received a red polycarbonate shelter and 2 cotton squares while mice all. Of the 13 labs that agreed to participate, 6 had a “good” in complex enrichment had a red polycarbonate mezzanine featuring compliance history and 7 had a “poor” compliance history. Labs with 2 side ladders to increase floor space and visibility, as well as crinkled good compliance were less likely to participate enthusiastically or paper for nest building. Bedding change caused an alteration of the complete the training (3/6 participated enthusiastically). In contrast, nocturnal activity pattern in all groups with animals held in standard labs with poor compliance history generally participated enrichment showing longer diurnal response to the procedure (P < enthusiastically (6/7 labs). Next, we explored whether forcing a lab 0.01). Females in standard enrichment revealed higher activity and with poor compliance history to participate affected enthusiasm. longer duration of the response to visual inspection (P < 0.01) while Surprisingly, we found that 2/2 labs with forced participation males in complex enrichment demonstrated fewer bout events and participated enthusiastically. Our preliminary observations suggest fight wounds (P < 0.05). Comparison of gender revealed males that the train the trainer program is a promising way to improve displayed less motility and shorter duration of the response to surgical outcomes in the labs that need most improvement, even if procedures than females (P < 0.01). Together our results suggest that participation is forced. environmental enrichment might influence the animal’s ability to cope with potential stressors and to generate variance of group P24 Performing a Canine Comprehensive Oral Health Assessment activity. Further exploration of noninvasive home cage motility and Treatment while Providing a Continuing Education combined with main physiological indicators might help us to assess Opportunity for Staff in a Research Setting the effectiveness of a new enrichment strategy.

EJ Powers*, RA Malbrue, LL Mattox P26 Evaluation of Effective Euthanasia Methods for Larval African Clawed Frogs (Xenopus laevis) University Laboratory Animal Resources, The Ohio State University, Columbus, OH IA Galex*1, CM Gallant1, LM Kuchenbrod1, C Fletcher1,2, AR Rogala1,2

Dental disease is recognized as a normal part of the aging process for 1Division of Comparative Medicine, University of North Carolina at dogs, and can negatively impact overall animal health, research, and Chapel Hill, Chapel Hill, NC; 2Department of Pathology and potentially cause pain and discomfort. A colony of beagles housed Laboratory Medicine, University of North Carolina at Chapel Hill, long-term on an IACUC-approved protocol, were noted to have Chapel Hill, NC developed dental disease. Common oral health examination findings revealed halitosis, gingivitis, tartar accumulation, and other dental African clawed frogs (Xenopus laevis) are a popular model for abnormalities. The clinical veterinarian used this as an opportunity to developmental biology and disease research. Ample published data perform a comprehensive oral health assessment and treatment are available describing effective euthanasia methods of adult X. (COHAT) for the dogs, while simultaneously offering RACE- laevis; however, evidence-based euthanasia recommendations have approved continuing education credits for participating staff. The not been published for X. laevis tadpoles. Recent publications have clinical veterinarian presented the CE in two components: (1) a shown that optimal euthanasia methods can vary greatly between presentation which outlined proper dental charting, tools, life-stages in other aquatic species with similar larval developmental techniques, and benefits associated with performing a COHAT and stages including zebrafish (Danio rerio). Therefore, the study aim of (2) a wet lab that provided staff the opportunity to anesthetize the was determine if larval X. laevis, specifically at various stages of lung dogs and perform a complete dental prophylaxis under general and gill functionality, require different euthanasia methods than anesthesia. The clinical veterinarian was present to provide guidance adults. We evaluated tricaine methanesulfonate (MS-222) and to attendees. Each participant rotated through the following roles: eugenol submersion, and rapid chilling as euthanasia methods for monitoring anesthesia, charting, and performing the steps of a tadpoles at 8, 23, and 45 d postfertilization. We completed an initial complete dental prophylaxis. Attendees were awarded 3 h of up/down pilot to determine the likely effective concentrations of continuing education credits. This CE provided a unique opportunity MS-222 and eugenol to be 6g/L and 600uL/L respectively. Tadpoles for staff to be directly involved in providing the gold standard of care were randomly selected for study groups (n=35) and placed in MS- for dogs maintained within our institution. 222 (6g/L), eugenol (600uL/L), a 1:5 ratio of ice:water, or a tank water control (n=20) for 15 min. Tadpoles were then placed in P25 The Effect of 2 Environmental Enrichment Strategies on recovery tanks of fresh system water for 4 h to evaluate the potential Wellbeing, Cage Activity, and Response to Common Husbandry for recovery. Heartbeats from a subset of 10 tadpoles from each group Procedures in C57bl/6crl Mice were assessed under a microscope prior to entry into recovery tanks and every 60 min thereafter. At the end of the 4 h, all tadpoles were F Scorrano*, D Lauras, F Cordoba observed for a heartbeat. Our results show that the MS-222 group had no detectable heartbeats after 4 h in recovery solution at all time Novartis International AG, Basel, Switzerland points tested. Eugenol at 600uL/L was variably effective at producing irreversible euthanasia; therefore, we deem eugenol at this Environmental enrichment is an important component of an animal concentration unacceptable for euthanasia, and will require further care program. It accommodates the innate behavioral needs of testing at higher concentrations. Although rapid chilling initially laboratory animals and enhances their wellbeing by giving them created a torporific state, it was ineffective at euthanizing animals at stress resilience to experimental manipulation and life in the all time points, thus, was not an acceptable method of euthanasia for vivarium. Behavioral, physiological, and biological parameters are larval X. laevis. Based on these data, we recommend MS-222 at 6g/L often used to evaluate the benefit of a new enrichment but little as an effective method for euthanizing larval X. laevis tadpoles of information is available on how different enrichment strategies might various life stages of development. impact group pattern of activity and response to common procedures. Increased home cage activity has been correlated with P27 Intratracheal Delivery of Human Tumor Cells in Mice under increased aggression in males and animal handling procedures such Continuous Isoflurane Anesthesia as cage change. These events during a period normally used for rest may lead to circadian disruption and have negative implications for J Hastings* welfare. Here, we measured physiological parameters and in-cage activity of group housed male and female C57BL/6Crl mice (n=72) Laboratory Animal Medicine, Janssen Research & Development, held in a standard and complex environmental enrichment for 42 LLC, Spring House, PA days and assessed their response, presented as activity recorded across cage electrodes, to recurrent interventions such as bedding We established a safe, reproducible survival procedure for the change and animal visual inspection. Mice with standard enrichment intratracheal (IT) delivery of human tumor cells and therapeutics

613 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 using (15 n=5 per group) Balb/C mice. Our investigation into this members compared to males in breeding groups (T = 4.0, P <.05). procedure was prompted by our oncology partners’ need for an These results suggest that adult male pigtails can be successfully orthotopic lung model. The procedure has numerous applications introduced to juvenile groups, which provides an additional because it permits the direct delivery of cells and therapeutics to the socialization option when no pair-mate is available. lower respiratory tract. We evaluated 3 methods of anesthetic delivery in support of the procedure: 1) injectable anesthesia 2) rapid, P30 Improved Practices on Gallus gallus domesticus Anesthesia shortterm isoflurane induction, and 3) continuous isoflurane Methods anesthesia. Continuous isoflurane was superior to the other methods because it allowed consistent atraumatic IT delivery with no J Lynn*, D Mead morbidity or mortality. IT instillation was performed via a modified IT stand, using visualization via an otoscope. The following results Animal Health Research Center, University of Georgia, Bogart, were achieved. The model has demonstrated safety, efficacy, Georgia reproducibility, and animal survival. This animal model will allow preclinical testing of agents that induce immunemodulatory effects in Current BSL-3Ag Gallus gallus (Gg) domesticus anesthesia methods, the lower respiratory tract. This can in turn lead to induction of for a terminal outcome, calls for isoflurane on a cotton ball in a systemic adaptive immunity and can be applied to other therapeutic conical tube placed around Gg domesticus’ beak until anesthetized. groups. The cotton ball holds the isoflurane in its liquid state so that spilling would not occur. In addition, the bird is placed on its back carefully P28 Forceful Perfusion of the Spinal Cord in Various Species so that the conical tube is maintained in an upright position. This is typically done with 2 technicians. This method was successful for JL Blauvelt*, N Ragland, M Baldwin, N Wiltshire, H Legato younger Leghorns. With adult Leghorns, having a fully developed cone and wattle, this method became problematic due to their head Comparative Medicine, University of South Florida, Palm Harbor, FL size and our inability to create a sufficient seal. Our team adapted the conical tube in a way that allowed the adult Gg domesticus’ head, Currently, several researchers require collection of the spinal cord cone, and wattle to be completely sealed within a funnel connected to and surrounding fluid for their rodent models. In the past, labor the isoflurane tube. The Leghorns were more susceptible to intensive blunt dissection of the boney structures surrounding the anesthesia using this method due to the reduction in evaporation of spinal cord were used. As a consequence, this method can damage liquid anesthesia typically caused by a poor seal. Not only did this tissue itself and prevents collection of spinal fluid. We have reduce the amount of time in sedating animals, but it also reduced implemented a method termed “forceful perfusion” that not only the amount of liquid anesthesia used during a study. We plan to try enhances efficiency, but can be used across several species to collect this way of anesthetizing Gg. domesticus to other birds of similar the entire spinal cord and spinal fluid simultaneously. Forceful anatomy, such as Broilers. perfusion uses readily available laboratory supplies that provides pressure using air, saline, or PBS through the spinal canal to dislodge P31 Refinement of Anesthesia and Analgesia for Rabbit Stereotaxic the spinal cord intact. Typically, spinal cord tissue is collected from Surgery the cervical vertebra thru lumbosacral vertebra with clean severing of the spinal nerves along the way. With forceful perfusion, our JJ Peirick*2, JJ Cooper1, FJ Sim1 researchers can effectively evaluate specific cell populations in the spinal fluid, capture the perfusate, and successfully rescue cells 1Pharmacology and Toxicology, State University of New York at needed for analysis. To date, we have found that using forceful Buffalo, Buffalo, NY; 2Laboratory Animal Facilities, State University perfusion not only decreases time, it enhances throughput, of New York at Buffalo, Buffalo, NY contributes to research aims and is proven successful in various species including mice, rats, gerbils, and juvenile guinea pigs. Provision of general anesthesia for rabbit stereotaxic surgery is challenging. Published descriptions are limited (injectable P29 Alternative Socialization Strategy for Adult Male Pigtail anesthetics, use of face masks). These methods have disadvantages. Macaques (Macaca Nemestrina) Aligning the rabbit’s head in the adaptor and the stereotaxic procedures can be lengthy. Short-acting, injectable anesthetics require J Cavallaro*, S Breaux, J Breaux multiple administrations, increasing complications (bradycardia, respiratory depression). Face masks do not allow for manual Veterinary Sciences, University of Louisiana at Lafayette, Lafayette, ventilations and are difficult to use with adaptors. While piloting a LA rabbit model of multiple sclerosis requiring injections of lysolecithin into the internal capsule, rabbits were anesthetized using refinements Nonhuman primate breeding colonies typically accrue excess adult to past descriptions. Anesthesia was induced in 12 New Zealand males when breeding using a harem strategy. These males are White rabbits via intramuscular injection of ketamine (35 mg/kg) commonly single housed when unable to locate a compatible partner. and xylazine (5 mg/kg). When rabbits were suitably anesthetized, When this occurred, we proposed housing adult male pigtail they were intubated after pre-oxygenation using a laryngoscope, size macaques (Macaca nemestrina) with juvenile conspecifics as an 3.0 cuffed endotracheal (ET) tube and an 8-French polypropylene alternative socialization paradigm. We introduced 3 different adult urinary catheter as a guide stylet. After intubation, a Bain circuit with males to 3 separate juvenile groups during 2016-2017. We observed a a fresh gas flow of 100% oxygen at 1 L/min was connected. total of 9 social groups twice/week, using instantaneous scan Following catheter placement, 0.05 mg/kg buprenorphine and 1.5 sampling on 3 juvenile/male groups (juvenile mean age=2.3 y; 30 h), mg/kg carprofen were administered separately via subcutaneous 2 juvenile only groups (mean age=2.16 y; 20.5 h), and 4 harem groups injections. Rabbits were then disconnected from the Bain circuit and (mean age 14 y; 32.75 h) and all occurrence focal sampling on adult carefully placed into the adaptor without ET tube disturbance. The males (n=7, mean age=13.3 y; 39.8 h). No differences in self-directed ET tube rested to the left of the upper incisors and was allowed to stereotypies (F = 1.06, P =.4), pacing (F=1.25,P =.2), or affiliative exit over the central bar of the U-frame on the left. The ET tube was behavior between juvenile, juvenile/male, and harem groups were reconnected to the Bain circuit, and isoflurane anesthesia was detected. While we observed elevated rates of play behavior among delivered between 0.5 and 3% throughout the procedure. juvenile groups over breeding groups (F = 55.39, P <.05), there was Glycopyrrolate was given IV if bradycardia developed. Spontaneous no difference between juveniles housed with males or peers only respirations were augmented by manual intermittent positive groups (P = .99). Comparatively, adult males housed with juveniles pressure ventilations (IPPV). Topical anesthetics were used prior to exhibited similar levels of pacing (T =.97, P=.36) and affiliative making the skin incision and drilling into the skull (0.25% behavior (T =.37, P =.72), but less time in close proximity to group bupivicaine, subcutaneously and 4% articaine, topically onto the

614 Abstracts of scientific papers 2019 AALAS National Meeting

skull). Rabbits were anesthetized for 3-4 h. Buprenorphine was P34 Quality Assurance: Ongoing Commitment to Animal Welfare readministered during recovery. Carprofen was continued at 1.5 mg/ with Emphasis on Reduced Regulatory Burden kg subcutaneously q 12 h on the day of surgery and for 3 postoperative d. There were no anesthetic deaths or serious J Kittel*, E Czarniak complications in any of the 12 rabbits. Animal Care and Use Office, University of Michigan, Ann Arbor, MI P32 Procedure Development for Repeated Infusion in Juvenile Nonhuman Primates Animal care and use programs are tasked with balancing a dedication to animal welfare and regulatory compliance with a JA Reynolds*, G Aylor, E Davidson, S Adual commitment to support research activities. A main objective of animal care and use programs is to address program oversight and Scientific Services, Altasciences, Marysville, WA instances of noncompliance. Common methods include the appointment of compliance officers or postapproval monitoring Restraining 48 juvenile nonhuman primtates (NHP) between personnel. However, these positions are often perceived by the 9-10-mo-old weekly for a 1-h long infusion while minimizing stress investigative community as policing forces and may contribute to factors required our team to adapt existing procedures for adult NHP investigator burden without any obvious benefit to the researchers and equipment where possible and develop new ones. The method themselves. Reducing the administrative burdens placed upon for infusion restraint included the use of a procedure cage where all investigators can free up resources and improve the quality and limbs were secured with limit movement. This method would be magnitude of research conducted without compromising animal highly stressful for animals under one 1 yof age. For previous welfare. Our IACUC has established a Quality Assurance (QA) team. infusion dosing of preweaned infants at 3 mo, the “snuggle wrap” The QA specialists work to facilitate compliance and minimize was developed to allow young animals to remain immobile while barriers to the conduct of sound, robust research. The program still feeling comforted by an hugging like sensation. Stress behaviors employs didactic visits in which researchers can openly share were noticeably less when using the snuggle wrap than when concerns, review their research protocols, and discuss the rules and manually holding as a restraint method. The snuggle wraps were expectations of the program without threat of consequence provided designed for ease of dosing vein accessibility while the remaining that there is no immediate threat to animal welfare. Visits are limbs are secure and comfortable. Knowing social pairs needed to conducted in a minimally disruptive manner, with the QA specialists remain in close proximity, the “snuggle board” was then developed. completing administrative tasks on behalf of the investigator Animals were then implanted with RFID chips, and the wrapped whenever possible. Results from the QA visits are bolstered by data juveniles were then labeled with temporary ID cards to allow for collection projects that assist in identifying programmatic easy identification once snuggled and placed on the board. These vulnerabilities. Select instances demonstrate how QA findings are boards hold 4 snuggled animals closely together which keeps social developed and used to direct programmatic change. The results of pairs in visual and auditory contact during dose administration. the QA initiative are improved compliance confidence for Following catheter placement, dosing limbs were secured with Velcro researchers, identification and assessment of programmatic issues, straps for dose duration. Animals were visually monitored, provided and reduced investigator burden. treats, juice, light projections, and chewing toys throughout dose duration by trained technicians. The snuggle board restraints were P35 Effect of the Analgesic Tramadol, Meloxicam, and Dipyrone on used for ~13 wk, until the animals were over 1 y of age and large the Behavior of Laboratory Rats enough for the procedure cage. JM Alemán Laporte*1,2, G Alvarado1,3, M de Souza Aranha Garcia P33 Just Hanging Out: Elevating Rat Enrichment in Small Spaces Gomes1, D Albert Zanatto1, M Bernardi4, AT Antorio Brasil1, CM Mori1 J Brekke*, JA Scholz 1Department of Pathology, University of Sao Paulo, Curridabat, Department of Comparative Medicine, Mayo Clinic, Rochester, MN Costa Rica; 2Laboratory of Biological Essays, University of Costa Rica, San Pedro de Montes de Oca, Costa Rica; 3Laboratory of Providing species-appropriate environmental enrichment is a critical Experimental and Comparative Pathology, University of Costa Rica, cog in the ever churning wheel of lab animal research. For rodents, San Pedro de Montes de Oca, Costa Rica; 4Environmental and finding new ways to provide enrichment without clogging up the Experimental Pathology, Paulista University, São Paulo, Brazil limited space of the housing cages can be a challenge. Our goal was to design an enrichment device for rats using the space between the Relieving the pain and suffering of laboratory animals is ethical and cage floor and the wire top that would engage the rat’s natural humane and it promotes good scientific practices. The inclusion of curiosity and propensity to climb. We developed a rat hammock pain suppressive agents in experimental procedures requires detailed using 3-inch diameter PVC pipe cut into 6 inch lengths and then cut research involving various analgesic or anesthetic options in order to in half lengthwise. The hammocks are hung using 16-gauge minimize experimental animal stress without negatively interfering galvanized steel formed into a hook. These materials are affordable, with the research results. We evaluated the effect of different durable, easy to remove for cage washing, and are sanitizable. We analgesics (meloxicam, dipyrone, tramadol, or the combination of piloted the device in a rat sentinel cage and observed both rats were tramadol with meloxicam or dipyrone) in the behavior of laboratory climbing onto the hammock within 2 min; after a week’s observation, rats. A total of 48 SPF male Wistar-Han rats, 8-12 wk old, were both rats were using the hammock to sleep, play, and groom during randomly divided into 6 groups (n=8 each): S: Saline- control group; various times of the day. Next, we expanded the number of M: treated with meloxicam; D: treated with dipyrone; T: treated with hammocks to 32 cages in 3 rat rooms. Observations were made 3 tramadol; MT: treated with meloxicam-tramadol; and DT: treated times per day during the light phase over a 2 wk-period to assess with dipyrone-tramadol. Animals received M: 1.5 mg/kg, D: 177.8 whether and how the hammocks were being used. We found that the mg/kg, and T: 17.8 mg/kg by intraperitoneal injection. Thirty hammocks were being used during 43% of the observations. The minutes after the injection, each animal was placed in a circular open hammock was primarily being used for lookout (67%) or sleeping field arena and the spontaneous behavior was recorded for 10 min. (16%), and less frequently for playing (9%) or grooming (9%). In Right after, the grooming transfer test was performed using a addition, the feedback from fellow animal care technicians has been fluorescent gel that was placed in the top of the head of each rat to overall positive. In conclusion, our observations indicate that the evaluate self-grooming at 2, 4, 8, 24, and 26 h after analgesic hammocks help to satisfy their natural curiosity and the urge to treatments; rating was determined based on scores of gel removal. climb. The hammocks have proven to be durable, easily sanitized, The data was analyzed using a 95% confidence interval considering and easy to place and remove. a P less than 0.05 as statistically significant. The D and DT groups

615 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 showed shorter distance traveled and lower average speed compared and Fridays (days 1, 3, 5, 8, 10, 12). Animals in groups 3, 4, and 5 with other groups. The DT group had a lower grooming transfer test (sham dosing) were given 1-2 mL of drinking water over 2 wk. Food score at all times evaluated, showing significant differences consumption and body weights were recorded on days 3, 5, 8, 10, 12, compared with other groups at 2 and 4 h. Group D also showed 15, 17, 19. In group 4 (days 1–5, 8–12 dosing) mean body weight was reduction in the self-grooming in the first 8 h, but it only had significantly (P < 0.05) lower than the control (group 1) on days 3, 6, significant difference compared with the control group (S) after 2 h of 8, 10, and 12. The body weights in this group began to trend upward the injection of the analgesic. To conclude, rats that were treated with after d 6 and were not different from control after d 12. Mean food dipyrone and dipyrone plus tramadol had more alteration in their consumption for the sham dosed groups (groups 3–5) were behavior. For this reason we do not recommend this analgesic in this significantly decreased on d 3. In groups 3 and 5, food consumption specific dose and administration route for studies where the recovered on d 5, indicating that the first dose causes measurable evaluation of behavior is important. effects that quickly resolve. The group 4 food consumption remained significantly lower than the control group until after d 13. In P36 Rectal Prolapse in Mice (Mus musculus): Using Nesting conclusion, body weight and food consumption data from group 4 Material as an Additional Indicator for the Timely Detection and demonstrate that sham dosing on a Monday–Friday schedule over 2 Study of Its Progression wk is sufficient to acclimate mice to the oral gavage procedure. This information is expected to help researchers to more efficiently J Poblete*, J Gimpel schedule gavage dose acclimations during weekends and holidays.

Pontificia Universidad Catolica de Chile, Santiago, Chile P38 Facilitating Access to Research Mice in China through Collaboration Rectal prolapse is defined as the exposure of the distal tissue of the rectum, appearing as a red mass of variable size protruding from the W McCarty*, K Lux, D Bannerman, C Miller anus. It is an animal welfare problem because it can cause pain, irritation, and infection, besides potentially affecting digestive The Jackson Laboratory, Sacramento, CA functionality of animals and being a possible sign of other anomalies (e.g. parasitism). It is a condition of relative frequency in aging In an effort to facilitate researcher access to quality mice in China and colonies making it sometimes difficult to strike a balance between accelerating worldwide discoveries to improve human health, we avoiding unnecessary suffering and deciding when to apply a partnered with several organizations and China customs to reduce humane endpoint, given the high scientific value that experimental the quarantine duration in Beijing, China. Collaborative relationships subjects have acquired at that age.Thus, the importance of early with institutions, vendors, and service providers in China promoted recognition to make the appropriate decisions about treatment or a team-learning environment benefiting all involved and allowing for humane endpoint. The degree of prolapse depends on the amount incorporation of U.S. laboratory animal health industry standards and condition of the exposed rectal tissue and the general condition with Chinese requirements. Bridging the language barrier through of the animal. Some scoring systems have been published to diagnose interpreters and hands-on demonstrations and on-the-job training rectal prolapse in mice. Our work proposes to add another sign: the required simplification of procedures while simultaneously seeking observation of white nest material (tissue paper). It is possible to operational efficiencies and maintaining quality. Dedicated U.S. team detect small exudate spots (or blood in advanced stages) thus members with operational responsibilities in China began pursuing allowing to decrease animal manipulation and restraint to verify the Chinese language classes to demonstrate our commitment to these state of the prolapse, sometimes even the need to open the cage, relationships and our mission of empowering the global biomedical which decreases the stress associated with these procedures. We will community in our shared quest to improve human health. The present the scoring method and decision tree that we have recently dedicated operational teams continue to work side-by-side with implemented in our vivaria. We believe that it is an improved tool to various Chinese organizations to provide resources, including hands- standardize criteria both to detect rectal prolapse and generate data on support and training. SOPs were written specifically for the on incidence and progression of this problem. This will allow us to quarantine facility and translated side-by-side in English and gather evidence on progression speed of rectal prolapse and, in this Chinese. Keeping AAALACi level of care in mind, areas of focus way, to refine decisions on appropriate times for the application of included animal handling and welfare, housing, shipping, quality humane endpoints. control measures, supplier management, autoclave validation, aseptic techniques, and sanitation methods. Documentation P37 Assessing Frequency and Duration of Oral Gavage Necessary including operational logs and training records were implemented to to Acclimate Mice document the work performed. Ultimately, the collaborative team approach demonstrated the integrity of the processes, thereby J Taylor*, JM Wilson highlighting the benefits of a reduction in the quarantine time needed, without compromising the health of laboratory mice in Laboratory Animal Medicine, Janssen Research and Development, China. Successful reduction of the quarantine period to 2 wk (50% Spring House, PA reduction) for mice arriving into Beijing promotes science, improves quality, shares knowledge, improves efficiencies, reduces costs, and In metabolism studies, body weight and food consumption are often allows for more timely use of the mice in experimental studies. measured data points. Undergoing oral gavage can result in decreased food consumption and weight loss in mice, affecting the P39 Evaluation of In-cage Filter Paper to Detect Murine Viral study results. To prevent this, investigators acclimate mice to oral Pathogens as a Replacement for Sentinel Mice gavage by sham dosing the animals for 1 wk prior to beginning the study, allowing for body weight to rebound prior to study start. KA O’Connell*1, GJ Tigyi2, C Murphy1, F Zeng1, DJ Hamilton1 However, the minimum frequency of dosing and duration of acclimation is unknown. Our goal was to determine the minimal 1Department of Comparative Medicine, University of Tennessee number of days of sham dosing necessary to acclimate mice to oral Health Science Center, Memphis, TN; 2Department of Physiology, gavage and to determine the time necessary for weight loss and food University of Tennessee Health Science Center, Memphis, TN consumption to recover. The study was conducted over a 3-wk period. C57BL/6J DIO mice were divided into 5 groups (8 male mice The replacement of the use of animals with an alternative technique per group): 1) control group that was not dosed or handled, 2) a is a fundamental principle within laboratory animal science. Recent handled group that was restrained daily but not dosed, 3) a group studies have evaluated alternatives to the use of live animals in dosed daily (days 1–14), 4) a group dosed Monday through Friday animal colony health monitoring using air plenums on ventilated (days 1–5, 8–12), and 5) a group dosed on Mondays, Wednesdays, racks, filter papers at the cage exhaust air ports, and filter papers on

616 Abstracts of scientific papers 2019 AALAS National Meeting

the lids of cages. While these methods have been shown to be Animal Nutrition and Physiology Team, National Institute of Animal effective for some types of infectious agents (e.g. parasites and Science, Wanju, Korea (the Republic of) bacteria) they have not been confirmed as reliable for other agents (e.g. several of the common viruses found in laboratory mouse Maximum residue limits (MRL) of pesticide in feed has been colonies) and the ability to collect samples can vary, dependent upon investigated to protect both the public health and livestock products. the design of the rack housing the colony animals. To determine a For establishing MRL in vivo pesticide residue experiment using reliable and simple alternative to sentinel animal testing, we livestock has been essential. However, livestock animals require a compared the use of filter papers placed directly on the inside floor special behavioral and social management including higher cost and of an empty mouse cage to the traditional use and testing of sentinel larger housing environment besides relative longer experiment mice in their ability to detect infectious agents transmitted by soiled periods. With that this study was conducted to evaluate the bedding transfer. Two commonly used cage filter materials were possibility that laboratory small animals can replace in vivo pesticide compared. The 3 experimental groups tested were: 1) filter paper residue experiments with livestock animals. First, 24 r pigs were type 1 located on the cage bottom, 2) filter paper type 2 located on the randomly assigned into 4 groups in which 0, 3, 9, and 30 mg/kg of cage bottom, and 3) 2, 10-wk-old, female, ICR sentinel mice. Each sulfoxaflor were supplied in feed. Next, 24 SD rats were randomly experimental group contained 7 replicate cages. Soiled bedding from assigned into 4 groups in which 0, 3, 9, and 30 mg/kg of sulfoxaflor pet store mice known to be shedding mouse hepatitis virus (MHV), were treated in the diet. After 4 wk, serum, muscle, fat, liver, kidney, murine Norovirus (MNV), minute virus of mice (MVM), mouse and small intestines were collected and sulfoxaflor residues were parvovirus (MPV), Theiler’s murine encephalomyelitis virus analyzed. Oral consumption of sulfoxaflor did not affect on the body (TMEV), and Helicobacter sp. was transferred to the experimental weight gain, feed intake, and feed conversion ratio, and accumulated cages weekly for 8 wk. Filter papers were evaluated at 1-mo and to tissues in the concentration dependent manner. Sulfoxaflor 2-mo time points by PCR. Sentinel mice were necropsied and tested residues in pig’s tissues were significantly correlated with those in by serology and PCR at the end of the 2-mo study. Results showed rats (P < 0.001). The model equations were formulated based on the that both types of filter paper reliably detected each of the infectious oral consumption of sulfoxaflor and its residues in tissues. Tissue agents shed by the study mice and were as effective as the use of live specific modeling equations were verified by investigating the sentinel mice. These results indicate that the use of filter papers similarity between calculated and measured sulfoxaflor residues. placed on the interior floor of mouse cages may be a reliable Moreover, the calculated and measured sulfoxaflor residues had technique to detect murine pathogens and serves as an alternative to more than 90% of similarity. In conclusion, sulfoxaflor residues in the use of live sentinel animal testing. rats and pigs were significantly correlated and the amount of sulfoxaflor residues in pigs could be estimated by the modeling P40 Behavioral Modification of an Undomesticated Pigeon equations. Therefore, we estimated the possibility that in vivo (Patagioenas leucocephala) Using Human Interaction pesticide residue experiments to set MRL could be performed using laboratory small animals instead of livestock animals that would S Glaspell*, N La Santa Medina, K Knapek, IM Washington contribute to reduce unnecessary animal experiments.

Office of Laboratory Animal Resources, West Virginia University, P42 Standardizing IACUC Protocol Review at an Institution with Morgantown, WV Multiple Species, over 400 Protocols, and 3 Campuses

White-crowned pigeons (Patagioenas leucocephala) are a species of bird K Ray* not often used in research, which may be related to their anxiety during normal human interactions, such as with cage changing. The Research Integrity, Oregon Heatlh & Science University, Beaverton, pigeon will often become stressed and flap around the cage causing OR feather loss and injury to itself. In order to improve the wellbeing of this pigeon, we decided to train it to help decrease its stress as well as The four research integrity specialists (RIS) at this institution manage improve its adoptability to various avian sanctuaries. A behavior over 400 IACUC protocols across 3 campuses. Species range from fish plan was designed to involve increased enrichment, housing space, and rodents to sheep and non-human primates. Our researchers use and human interaction, as well as audio nature sounds. Two handlers animal models to study a broad range of health-related topics, work daily with the pigeon, in increasing time increments over including vaccine development, cancer research, cardiometabolic, several months. The bird was scored daily, 5 d per week following a and neurologic disorders. Over time the vastness of the program led rubric which included observation of the pigeon, behavior of the to inconsistency in how IACUC protocols are reviewed by the RIS. pigeon while the handler was approaching the cage, behavior of the For the past 2 years, RIS have worked with assistant research pigeon while receiving auditory cues from the handler, reaction to integrity officers from IACUC and IBC to develop a standard review movements of the handler, and tracking of negative stereotypical checklist that applies broadly to all IACUC protocols at this behaviors, such as rousing and wing flapping. Fecal samples were institution. This checklist follows the format of the electronic eIACUC collected throughout training to evaluate cortisol levels with protocol form and can be used when reviewing initial protocols, behavioral modification program. We determined that the bird modifications, and 3-year renewals. Implemented in early 2019, the responds best to a slow approach of hands when they are below the review checklist has been successfully integrated into office processes bottom half of his body, but we saw more anxiety and stress when and provides a standardized and organized approach to protocol the handler performs quick, unexpected movements. Since following review. It is a living document that will evolve over time with an outlined training plan a decrease in self-injurious behavior as well changes to the regulations and our animal care and research as decreased rousing and wing flapping was observed. A step-wise program, and will continue to provide a solid foundation of increase in nonhandling human interaction can be used successfully consistency in IACUC protocol review for current and future RIS. for behavioral modification of an undomesticated avian species to improve pigeon welfare and reduce the risk of injury to the bird and P43 Using Nonanimal Tools to Supplement a Training Program staff. This tool can be used by other institutions attempting to rehabilitate undomesticated species for transfer to an avian facility. KA Blanchette*, K Cough

P41 Study on Evaluation of Sulfoxaflor Residues in Pig Tissues Animal Welfare and Compliance, The Jackson Laboratory, Bar Using Rats as an Animal Model: Reduction and Replacement to Harbor, ME Enhance Animal Welfare As members of the laboratory animal science community, we are K Kim*, J Chun, s Ji, y Lee, J Jeong, H Bang, M Kim, S Lee constantly looking for alternatives to animal usage whenever possible. We have worked to develop training tools using 3D

617 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 modeling that reduce the number of mice used but meet the number of facilities that are starting to socially house monkeys with standards for a strong training program. Using 3D modeling, we these kinds of implants. There are fewer reports of social housing have created a training tool for body condition scoring (BC1: monkeys with chronic catheters. We recently pair housed 2 female emaciated through BCS 5: obese) in mice. We are in the process of cynomolgus macaques (Macaca fascicularis), 1 of which had a chronic developing additional models. One that mimics malocclusion and is catheter and catheter protection system. The monkeys had been used for teeth trim training and another that mimics a pregnant dam paired prior to the catherization and were highly compatible (e.g., and is used for pregnant female palpation training. We hope to displayed a great deal of prosocial behavior). Monkeys were briefly implement these newer models in the next few months and share separated after surgery. Behavioral and clinical technicians remotely them with the research community. The use of nonanimal tools to monitored the monkeys as they were reintroduced. The monkeys supplement training is heavily supported at our organization. The groomed each other and engaged in other prosocial behaviors, and models already in place have received high praise and we have been we did not notice any signs of aggression or excessive manipulation encouraged to continue the development of new tools that can be of the catheter protection system. Further, there were no adverse shared with the laboratory animal science community. Our clinical effects that resulted from the social housing. While this may inventions help to decrease mouse usage while still maintaining a not be feasible in all circumstances, pair housing animals with high level training program. catheter protection systems is possible, and is a refinement for those animals. P44 Caretaker Attitudes towards Naming Animals In Research Facilities P46 Nonhuman Retirement from a Pharmaceutical Company: Guidelines and Challenges EJ White*, K Coleman LE Shulder*, MC Kundu Oregon National Primate Research Center, Beaverton, OR Veterinary Sciences, Bristol-Myers Squibb, New Brunswick, NJ One important outcome of high-quality animal care is the close relationship that often develops between caretakers and animals. Many institutions have well-established animal rehoming programs Because of this relationship, care staff often name the animals, a for a variety of species, such as canines, rodents, rabbits, and fish. practice that is not universally accepted. It has been suggested that However, the retirement of nonhuman primates is often a more names help staff identify individuals and strengthen the animal- complex decision. The question of which animals are selected for caretaker bond thus improving care. However, no work has been retirement, where they will live out their retirement, and funding for done to specifically examine the role that naming plays in animal their lifetime care are just a few of topics that need to be discussed. care or, important, the effect it has on the care staff. In this study, we The 3Rs must be considered in the toxicology environment prior to examined this question through a questionnaire. The survey included any animal rehoming or retirement. Animals that have an acceptable questions regarding if and why people named animals, as well as research and clinical history, as determined by a veterinarian, and are their opinions on naming animal subjects using a 5-point likert scale no longer necessary for the facility’s research protocols can be (from strongly disagree to strongly agree). Seventy-eight people considered eligible. Sanctuaries must be evaluated in-depth to assure responded to the survey, including husbandry, clinical, and research adequate care, excellent animal welfare, and security in terms of technicians; veterinarians; and managers, all of whom worked with institutional privacy. Discussion about sanctuary requirements, such monkeys (some also worked with rodents). The vast majority of as bloodwork or sterilization procedures, and logistics and pricing of respondents (87%) reported that they named research animals. Of relocating these animals to their new retirement home should be these, 46% reported that they would name rodents/rabbits in done early in the process. Once sanctuary vetting is completed, it is addition to monkeys. Eighty-one percent reported that naming necessary to understand the need for lifetime support of the retired improves the bond with the animals. Opinions about how naming primates. The retirement of nonhuman primates comes with a unique affects animal care varied. Most respondents (53%) reported that they set of hurdles that may need approval at many levels of the somewhat or strongly agreed with the statement “I pay more institution. At our facility, we used our security department to run a attention to named rather than unnamed animals.” Interestingly, 76% background check on the chosen sanctuary to ensure there were no disagreed with the statement “I provide better care to named strong connections to antiresearch organizations. We have kept in animals,” presumably because they felt they provide excellent care to close touch with the sanctuary in order to continue to provide all animals. Our survey also revealed that naming can have support and for the possibility of future use for additional animals. emotional consequences. Almost half of the respondents (49%) Support outside of funding can include donation of equipment, agreed with statement “I am more emotional when a named, as volunteering, or medical advice for the animals. Preparing guidelines opposed to unnamed, animal has to be humanely euthanized.” and understanding challenges is the first step to set-up a primate Knowing this information can help facilities better understand the retirement program for success. animal-human bond and what that means to animal care as well as human wellbeing. P47 The Use of Modified Rodent Vascular Access Buttons to Refine Repeat Blood Sample Collection in Miniature Pigs P45 Pair-housing Macaques with Chronic Catheters LE Brierley* J Shelton*1, J Stanton1, H Price1, M Robertson-LeVay1, H Wu2,1, J Sacha2,1, K Coleman1 Frontage Laboratories, Inc., Concord, OH

1Oregon National Primate Research Cener, Beaverton, OR; 2Vaccine Miniature pigs are becoming an increasingly popular species in and Gene Therapy Institute, Beaverton, OR pharmacology and toxicology. While there are many reasons that make miniature pigs good models for this research, there are also Social housing is known to be a highly effective form of enrichment some challenges. One of the challenges is that pigs quickly develop for macaques in research facilities. However, facilities are often an aversion to the repeat blood sample collections that can be hesitant to socially house monkeys that have implanted devices, such stressful to the animals and staff. To facilitate sample collection, as cranial implants or chronic intravenous catheters, to avoid many facilities place vascular access devices such as external jugular excessive manipulation of the devices by the partner. In these catheters or vascular access ports. The use of modified rodent situations, pair-housed monkeys are typically separated from their vascular buttons for blood sample collection in miniature pigs is partner, which may be stressful for both individuals. Researchers becoming more common in Europe because of the ease of access, have shown that pair housing does not interfere with cranial reduced stress for animals and staff, and cost. The use of these implants for compatible macaques, which has led to an increasing buttons has been slower to catch on in the U.S. due to concern for

618 Abstracts of scientific papers 2019 AALAS National Meeting

potential infection. In February 2019, we placed our first modified threshold established in literature indicating transfer of protective rodent button in Göttingen miniature pigs (Sus scrofa domesticus). passive immunity. The premature lambs (n=5) were given a plasma These buttons were modified to have an extra magnet to hold the transfusion (20mL/kg) 24 h after delivery onto the artificial placenta button cap in place. In the past few months we have seen low rates of machine. Blood samples were taken from the dam at the time of infection (0%) in the few animals (n=4) with buttons placed. Staff surgery and from the lamb at time points both before and after the members gave direct feedback that they prefer collecting samples transfusion. Serum IgG levels were determined via an ELISA. Our from miniature pigs with buttons because of the ease of sample data indicate that there was an initial increase in the lambs’ IgG collection and the animals’ lack of aversion to sample collection. We concentrations measured 1 h after the plasma transfusion. However, will present evidence that the use of modified vascular access buttons this was typically decreased by 24 h posttransfusion. In addition, in may be a refinement for miniature pig blood collection for an effort to improve dam IgG levels, a vaccination program was pharmacological and toxicological research. implemented and the effects were analyzed. Overall, a single intravenous plasma transfusion was capable of raising serum IgG P48 The Use of Themed Enrichment Activities to Engage levels in the premature lamb, but these levels were not elevated to Employees in Animal Enrichment Activities the threshold of being considered protective. Future work is necessary to investigate the effects of multiple transfusions over the LE Brierley* course of the first days of life.

Frontage Laboratories, Inc., Concord, OH P50 Development of a Quality Assurance Hideaway Program as an Adjunct to Technician Training and Facility Oversight Animal enrichment programs are an important part of any animal research facility, but for an animal enrichment program to really excel LG Williford*, C Inman it takes time and dedication from animal care staff. We recently instituted monthly themed animal and staff enrichment activities Lab Animal Care Unit, University of Tennessee, Memphis, TN with the goal of providing additional enrichment for our animals and engaging our animal care staff in our enrichment program. After the Having the ability to evaluate the efficiency of laboratory animal first event, it was apparent that our goals of engaging technicians in technicians throughout an animal facility is difficult because you enrichment activities was met as technicians starting making cannot be in the animal rooms or facilities with them at all times to suggestions about types of activities and enrichment that could be evaluate their work. Knowing how to structure training for certain done. The immediate outcome that we did not expect was the procedures and policies or customizing different training aspects for involvement of nonanimal care staff members to become involved in a technician can be a challenge when you are not certain what areas our enrichment program as rumor of these activities spread and of training need the most attention. To help in the evaluation of people started asking if they could join our events. This triggered a employee performance and adherence to established policies and dialogue with nonvivarium workers about what our animal research procedures, a quality assurance (QA) hideaway program was entailed and the level of care that we provide to our animals. developed. The QA hideaway program consists of placing plastic Ultimately, we decided to expand these events to include mice in empty mouse boxes or QA cards throughout the facilities for nonvivarium staff and use them as an opportunity to engage and the technicians to find and report to the veterinary staff, as they educate all of the employees at our site about animal research and would normally for a live mouse, or by following the instructions on animal welfare at our facility. Themed enrichment activities were the QA card. A play money reward system was also created and provided for a range of species (rats, mice, canines, miniature pigs, technicians would receive “cash” prizes for finding the QA hideaway and cynomolgus macaques) and typically involved the creation of item. The sooner a QA mouse or card was found after being placed, destructible enrichment item or food enrichment coupled with either the greater the prize. Technicians can then purchase real prizes using auditory or visual enrichment. their play money. When the program started, 46% of technicians found their QA mice/card within 24 h. Within 48 h, 92% of P49 Effects of a Single Intravenous Plasma Transfusion in a technicians had reported their QA hideaway. By 96 h, 100% of QA Premature Lamb (Ovis aries) hideaways had been reported. With these findings we were able to identify technicians that need retraining on some procedures and L Krueger*1, JS McLeod2, E Sarosi2, C Fry1, R Griffin3, PS Allen1, A policies and how to structure the training for the individual or group. Rojas-Pena2, RH Bartlett2, J Nemzek1, G Mychaliska2,4 The QA hideaway program is a fun and effective way to monitor technician performance and training needs, while providing a 1Unit for Laboratory Animal Medicine, University of Michigan, Ann positive reward system at the same time. Our goal is to increase Arbor, MI; 2Extracorporeal Life Support Laboratory, Department of reporting results so that >90% of hideaways are reported within 24 h. Surgery, Michigan Medicine, Ann Arbor, MI; 3College of Veterinary Medicine, Michigan State University, East Lansing, MI; 4Fetal P51 Prioritizing Animal Comfort and MInimizing Restraint and Diagnosis and Treatment Center, C.S. Mott Children’s Hospital, Handling Stress in Larger Nontraditional Species Michigan Medicine, Ann Arbor, MI LM Panepinto* As a model for determining the effect of extracorporeal membrane oxygenation on fetal circulation, surgery, and treatment, premature P&S Farms, Masonville, CO lambs (118-121 d) are delivered and placed on an artificial placenta machine. Neonates are one of the most vulnerable age groups for One of the greatest challenges for researchers, veterinarians, and morbidity and mortality due to infections. Some species, like animal care staff is finding user-friendly, humane methods for humans, are born with passive immunity to pathogens, having handling and restraining some larger, nontraditional species. Young received maternal immunoglobulin (Ig) transplacentally during domestic swine, minipigs, and goats may be some of the more gestation. In contrast, other species, like newborn lambs (Ovis aries), difficult animals to handle, especially for procedures like blood are not born with protective levels of circulating maternal Ig and sampling, IV infustions, oral dosing, and hoof trimming. We descibes derive the majority of their protection from consuming Ig-rich a user-friendly, portable fold-up device for humanely and easily colostrum within the first few hours of life. With the goal of restraining small- to medium-sized animals, including dogs, goats, increasing survival and improving welfare of the premature lamb, pigs, sheep, and small calves with a minimum of stress to animals we sought to determine the effects of a single intravenous plasma and handlers as well. The animals are comfortably restrained in transfusion on its immune status. Our hypothesis was that sternal recumbancy, supported by an acrylic, fully washable fleece- administering plasma harvested from the dam to premature lambs lined hammock attached to a light weight folding frame. This after delivery would increase Ig levels above the 1500 mg/dL portable frame has a small footprint, can be stored in a closet or other

619 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 storage area, and can easily be transported for use at alternate was no intervention outside of standard breeding protocols locations within the research or clinical facility or elsewhere. It also (enrichment, food, etc.). Each cage type ran 8 cages of C57Bl/6J facilitates veterinary examinations and treatments, especially when breeding pairs, and pups were kept in the same caging system. The the avoidance of anesthesia or chemical sedation is important. An second litter from each cage was kept for 6 mo. Parameters of alternative model does not fold up but uses variably sized hammocks temperature, relative humidity, and ammonia were collected to humanely hold larger animals on a sturdy steel frame. Most weekly to determine the cage environment. Variables of litter size, animals comfortably restrained using this technique adapted from pup loss and respiratory tissue pathology were collected to our original crank up sling for larger animals, readily acclimate with determine effectiveness of breeding and health of the animals. Litter minimal or no training, and often fall asleep during procedures. This size was counted on d 1, 21, 28, and at 6 mo. Respiratory tissues were innovative methodology provides a portable, humane, user-friendly collected from pups at 21 days and 6 months, as well as tissue from alternative to some of the more stressful, traditional approaches to mothers at 6 months after last litter, respectively. The novel cage handling and restraint. design saw an advantage over the traditional design in how many pups make weaning age. The novel design also saw significantly P52 It Takes a Village: Improving Welfare with a Team Approach lower pup loss (9%) than the traditional design (27%). The traditional cage saw significantly higher temperatures than the novel cage, also L Bader*1, M Janavaris1, T Bodvarsdottir2, JJ Hansen2, K Coleman1, P associated with the day of measurement. Humidity within cages was Kievit1 not significantly different. Both cage types saw increased ammonia over time. The novel cage saw similar ammonia levels on d 21 as the 1Oregon National Primate Research Center, Beaverton, OR; 2Novo traditional cage on d 14, which is remarkable given the cage change Nordisk, Bagsvaerd, Denmark schedule. The improved weaning result in the novel cage design are thought to be a result of both reduced handling and reduced airflow There are many factors that need to be considered when caring for inside the cage. groups of research animals, including clinical, behavioral, and scientific issues. Often, decisions are made without taking all of these P54 Comparative Behavior and Cortisol Levels in the Rabbits needs into account. We use a team approach to address welfare needs (Oryctolagus cuniculus) Anesthetized by the Use of Masking or of animals. Members of clinical, behavioral, and husbandry staff Premedication meet regularly to discuss issues surrounding individual animals. This increased communication has greatly improved animal care. LE Ramagli*1, DR Hill1, DJ Anderson2, D Norton1 Recently, we took this process a step further by applying it to animals even before they arrived at our center. Twenty-six male Mauritius 1Department of Veterinary Medicine , MD Anderson Cancer Center, cynomolgus macaques (Macaca fascicularis) arrived and were housed Missouri city, TX; 2Agricultural Sciences, Sam Houston State in 2 groups in indoor pen housing. These monkeys were brought to University, Huntsville, TX our facility as models for pharmaceutical therapeutics for obesity. Because we planned to have these animals on studies for several Anesthetic procedures tend to induce stress in laboratory animals. years, we wanted to anticipate and avoid as many issues as we could. Reducing this stress is not only imperative to their overall wellbeing, Prior to the monkeys’ arrival, we sought input on housing and but also ensures quality patients for researchers. Improving methods clinical and behavioral parameters from a large team of people in for anesthetizing animals allows for a smoother induction, procedure, these areas. For example, due to the complexity of maintaining recovery, and postoperative experience for not only the animal, but group-housed animals, project staff includes behaviorists who work the technologist caring for the animal. In this study, we specifically closely with researchers. There are monthly meetings with behavioral focused on the rabbit model, as this species tends to exhibit large management staff to discuss issues such as group dynamics, how amounts of stress in most handling circumstances due to their nature animals respond to various enrichment items, and how best to track as a prey animal. Despite this fact, no definitive study has individual behavior. This team approach fosters engagement among determined whether anesthetic premedication reduces stress versus people involved, which has, in turn, increased buy-in for novel mask induction alone. The objective of this pilot study in adolescent enrichment and housing options. Further, this process has opened rabbits (n=12), was to comparatively measure anxiety indicators to honest communication among all parties; staff feel empowered to determine the effect on stress. To do this, blood cortisol levels, as well make suggestions and provide valuable feedback. This team as observed behavior, were recorded after the animal has been approach has elevated welfare for these animals and is a refinement anesthetized with 1 of the 2 previously described methods. Half of to animal care. Importantly, this approach has set the project and the the rabbits were masked down with isoflurane only, once they were animals up for success for years to come. anesthetized, blood was collected immediately after induction. Additional rabbits were premedicated prior to being masked for a P53 Can Cage Design Affect Breeding Performance? procedure, and blood was collected using the same method. While most samples were within normal range for both groups, 2 of the L Kramer*1, DR Totman2, B Burke2, S Soprano2, EK Daugherity2 higher cortisol levels were in the masked group. Rabbits that were in the masking group tended to back away forcefully from the mask, 1Engineering, Lenderking Caging Products, Millersville, MD; 2CARE, hold their breath, and occasionally escape restraint and try to flee. In Cornell University, Ithaca, NY contrast, rabbits who were premedicated before their masking appeared completely relaxed and did not resist. While cortisol levels Individually ventilated caging (IVC) is used in many animal facilities. were fairly inconclusive, the behavior suggests a lower stress Standard husbandry practices in this type of equipment are a 14-d response in the premedicated group. While this study is a good cage change schedule and airflow rates of 60 air changes per hour starting point in the right direction in determining improved (ACH) through each cage. There is a novel cage design that employs methods for anesthetizing animals in a stress free manner further a perforated false bottom to hold the bedding and air percolates investigation is warranted. through the bedding at lower rates. This is a divergence from traditional IVCs where bedding is placed on a solid bottom and air P55 Employing the 3Rs to Surgical-skill Training passes over top of the bedding. The novel cage design had previously been tested in a 5-mouse setting, to operate on a 21-d cage change L Denning*, S Lownie schedule using airflow rates of 30 ACH. These changes were hypothesized to be advantageous to the breeding scenario. A CNS, University of Western Ontario, London, Canada traditional IVC design was compared to the novel IVC design in breeding C57Bl/6J mice on manufacturer recommended settings. A neurosurgical laboratory was developed to train second-year Breeding pairs were bred undisturbed for 6 months, meaning there neurosurgery residents on microsurgical techniques, while seeking to

620 Abstracts of scientific papers 2019 AALAS National Meeting

embrace the goals of the 3Rs.Students are trained in microsurgical P57 Openness in Animal Research: A Pharmaceutical Company’s dissecting and vascular repair in order to improve their microsurgical Global Approach skills for the other neurosurgical tasks such as brain tumor resection, carotid plaque removal, aneurysms, and hemorrhage control among M Gallacher*1, S Robinson2, C Traher3, S Östman4, D Goldsteen5, L other brain operations. Each second-year neurosurgery resident Martoft4, PL Finnemore6 attends one half-day session twice a month. The training is divided into 5 modules. Module1 consists of observing training videos. 1Animal Sciences and Technologies, Clinical Pharmacology and Module 2 comprises the duck wing, vascular dissection, and Safety Sciences, R&D, AstraZeneca, Boston, MA; 2Animal Sciences anastomosis. Modules 3,4 and 5 introduce the live rat under general and Technologies, Clinical Pharmacology and Safety Sciences, R&D, anesthesia for femoral artery, vein and end to side dissection, AstraZeneca, Alderley Park, United Kingdom; 3Animal Sciences and clipping, and transection and end to end anastomosis. These modules Technologies, Clinical Pharmacology and Safety Sciences, R&D, are each repeated 5 times consecutively. Simulators, phantoms, and AstraZeneca, Cambridge, United Kingdom; 4Animal Sciences and the brachial vessels of the duck wing are a good starting point for Technologies, Clinical Pharmacology and Safety Sciences, R&D, microsurgical dissection, and replaces the need for a live animal AstraZeneca, Gothenburg, Sweden; 5Animal Sciences and model. Rats have been instrumental as a surgical training model. Technologies, Clinical Pharmacology and Safety Sciences, R&D, Neurosurgery residents and fellows dissect the rat femoral vessels to AstraZeneca, Gaithersburg, MD; 6Global Animal Research Assurance, learn anastomotic techniques. We have replaced commercially Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, purchased rats by obtaining old breeders, and rats which have Cambridge, United Kingdom reached their study endpoints from fellow researchers. The optimum utilization of our rats is via bilateral vessels on the same animal, Animal research is a topic where there is limited awareness and cutting our rat numbers down by half. By adapting new methods of many misconceptions. Many organizations have core values that training using phantoms and biological models, we are able to capture the spirit of “doing the right thing,” but representing those improve upon the resident skills necessary for neurosurgical actions accurately is key to earning the public trust. Gaining society’s techniques using the live anesthetized rat and progressing toward acceptance of animal research is imperative for life science the clinical patient setting with neurosurgeon mentorship. researcher’s ability to operate. The Council for Science and Animal Competency-based training has been mandated by Royal College of Welfare (C-SAW) is our expert group leading a global approach to the Physicians and Surgeons of Canada for the training of surgeons in all use of animals in research, including the promotion of initiatives specialties including neurosurgery. Here. efforts are made to around openness and transparency. The Animal Sciences and incorporate the 3Rs into this training protocol and improvements are Technologies (AST) department support all internal animal research, continually sought. local compliance, champion the 3Rs and animal welfare, and are core members of C-SAW. AST have developed programs describing our P56 The 3Rs in Oncology Research Training: Creation and Utility animal use in transparent ways. Understanding the messages to of a Phantom Training Tool for Acquiring Proficiency in Calipered deliver and the intended target audiences, AST has created multiple Tumor Measures mediums to educate and answer questions regarding animal use. Awareness days, poster sessions, information tables, print and social MS Metzler*, J Vineyard, L Busbin, D Manning, CA Parks media information, vivarium tours and short films are all effective methods to communicate our ways of working with animals. Sharing Charles River Laboratories, Charlotte, NC our animal work with nonanimal users has been well received. We have seen an increased interest in our internal openness events The 3Rs (replacement, reduction, and refinement) represent a resulting in greater participation. We recognize the value these events practical method for responsible and humane care and use of animals provide for the AST staff to share their expertise and passion and that in accordance with the principles of the Guide. The Guide also a byproduct of these openness events is a powerful motivator for stipulates “all personnel involved with the care and use of animals them. It is clear to the authors that we need to do more to share must be adequately educated, trained, and/or qualified in basic animal research information with a wider audience. Sharing our principles of laboratory animal science to help ensure high-quality openness initiatives globally allows our AST sites to learn from each science and animal wellbeing.” Combining these principles has other and celebrate our successes. A global approach to the resulted in the creation of phantom training tools aimed at replacing responsible use of animals in research and openly sharing the use of live animals for training purposes. By enabling trainees to information about our animal work is an important step to earning attain proficiency in handling and technique prior to live animal the public’s trust. training and study work, phantom use also promotes reduction and refinement. Despite the options for phantom rat and mouse training P58 Efficacy of Dog Appeasing Pheromone in Reducing Anxiety in tools for handling, injections, feeding, and blood collections, there is Laboratory Beagles not an option available for the practice of measuring subcutaneous tumors—a mainstay of oncology xenograft research for the screening M Bulthouse* of chemotherapeutic in vivo efficacy. Therefore, to train technicians to accurately measure subcutaneous implanted tumors, we created our Surgery and Efficacy, Charles River Labs, Mattawan, MI own phantom training tool using hot glue tumors and commercially available indoor hunting cat food feeder mice. We used a hot glue Dog appeasing pheromone originates from intermammary sebaceous gun to create tumors of variable size and shape which we wrapped in glands of lactating bitches shortly after whelping; it calms and craft batting to create defined tumors with a slightly amorphous reassures the offspring. Several studies have shown that DAP has a shape. To create the phantom mouse, we wrapped a piece of felt calming effect in canines in a variety of situations. However, there secured with double sided tape to the hard-plastic mouse body. The has been little to no research on how DAP affects laboratory canines. mouse “skin” (nylon/rayon fibers) was replaced, and tumors Sling acclimation is a stressful procedure that occurs often in “implanted” under the “skin” on top of the felt. The mouse could laboratory studies. We evaluated if DAP is successful in reducing then be grasped by the scruff behind the ears and tumors measured stress in laboratory beagles during sling acclimation. This was with calipers. Additional options include opting out of wrapping the evaluated using a general behavioral analysis scoring, heart rate, tumors in the batting for beginner training and then advancing the respiratory rate, white blood cell differentials, and serum cortisol training with smaller tumors and more batting material. This tool levels. Beagles were placed in the sling for 30 min; heart rates and was used to effectively train technicians prior to in vivo acquisition respiratory rates were evaluated in the first and last 5 min of sling of calipered tumor measures and demonstrates application of the acclimation. General behavior analysis was evaluated every 10 min 3Rs. while the beagles were in the slings. Adaptil, a widely available form of DAP comes in sprays and diffusers. Sprays, diffusers, and a

621 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 combination of both were evaluated in sixteen male beagles in postures and behaviors unknown to technical staff, but normally 4groups: 1 control, 2 spray only, 3 diffuser only, and 4 spray and expected for rabbits. Pens saved time in terms of cleaning by diffuser. Diffusers were placed in rooms for 30 d and at least 24 h allowing technicians more time to assess and consolidate human- prior to data collection. Sprays were applied to towels and placed in rabbit interactions. Technicians were trained to assess some welfare cages with the dogs at least 15 min prior to the stressful situation. indicators such as docility in terms of ease of handling, postures Blood collection took place on Wednesdays whereas sling acclimation (normal versus abnormal), fear, presence or lack of aggression, took place on Fridays for 6 consecutive wk. The first 2 wk data washing/grooming, mutual sniffing, and identification of marking established pretest values, while the last 4 wk were used to evaluate spots. Technicians often choose to sit on the pen floor letting rabbits Adaptil. It was hypothesized that DAP will reduce anxiety in come close, gain confidence, and lose their natural prey-fear. laboratory beagles during sling acclimation. Results showed that Technicians may play with the rabbits. While no biologic DAP successfully reduces heart rates and respiratory rates during experimental data was collected, staff summarizes that the animal sling acclimation and helped to reduce behaviors associated with care staff participation in learning and assessing during this process stress. Eye contact, lip licking, panting, and destructiveness were all provided them opportunity to get involved with improving rabbits’ shown to have a statistically significant P value per protocol. White welfare by strengthening human-rabbit interactions. blood cell differentials and cortisol levels were inconclusive. In the future, better blood biomarkers could be evaluated for stress. Blood P61 Optimized Methods and Refinement Strategies for Dosing and could also be collected during/toward the end of sling acclimation as Blood Sampling in Rodents to Enable 3Rs Impact it seems colony beagles are content in their home cages and during blood collection. S Oglesby*2, P Ebert3, NA Bratcher1

P59 Use of an Automated Peritome for Nonhuman Primate Dental 1Animal Welfare & Compliance, AbbVie, Chicago, IL; 2Toxicology, Extractions AbbVie, North Chicago, IL; 3Comparative Medicine, AbbVie, North Chicago, IL M OBrien*, R Zweig, H Sidener We are committed to using and developing protocols that reduce the Oregon National Primate Research Center, Beaverton, OR number of animals required and refining procedures to minimize pain and distress. Improvements in analytical methods enable drug- Our population of approximately 5,000 NHPs, 12% who are over 12 y level analysis from smaller blood volumes with interim drug level of age, not infrequently require dental extractions to be performed. determination by serial microsampling. Serial microsampling reduces Our certified veterinary technicians are trained to extract all types of the number of rodents (specifically mice) required, costs, and teeth, including canines, which is a labor-intensive process with an resources over terminal composite sampling. We established a global ergonomic risk for repetitive motion hand injuries. With an average guideline to align expectations and to ensure optimal refinement of of 2-4 dental extractions per week, we have explored ways to refine methods used for microsampling. Under this guidance, consideration our tooth extraction processes to reduce exposure risks, alleviate is given to route of dosing and site of venipuncture, as different ergonomic concerns, and improve efficiency and patient outcomes. A venipuncture sites may yield differences in clinical pathology values. technique using an automated periotome was identified. It provides If performed accurately, collection of blood from the lateral tail and a rapid in and out mechanical action using a thin blade to saphenous veins are preferred sites for blood collection as these atraumatically sever the periodontal ligament. This approach methods allow serial collection with minimal venipunctures. eliminates the need to remove alveolar bone prior to extraction and However, the site of venipuncture should differ from site of drastically reduces procedure times. Reduction of the use of elevators intravenous (IV) drug administration due to potential extravasation, and drills effectively removed the components of the process or leakage of the infused substance into the extravascular tissue typically associated with exposures. Technicians report that while around the injection site. For IV dosing into the lateral tail vein, the there is a learning curve to using this new equipment, the reduction saphenous vein can be used for blood collection, but is not always of the twisting action and pressure needed for manual luxation has the preferred technique of technicians. Sampling from the greatly decreased ergonomic concerns. Using this technique, we have submandibular or jaw vein is not optimal as it has been shown to improved animal welfare by reducing the time a NHP requires cause tissue damage and could cause undue pain and distress since inhaled anesthesia while decreasing the trauma that comes secondary the facial nerves are near the vascular bundle. Therefore, we needed to extracting teeth using more invasive techniques. to identify an alternative IV dosing site. Anesthetized penile vein dosing provides a refinement for IV dosing leaving the lateral tail P60 Retrospective Assessment of Rabbit Housing from Individual and saphenous veins available for serial blood collection and can be Cages to Floor Pens used in a variety of pigmented mouse strains. Here we will present how AbbVie has implemented our global guideline and this refined MM Ricca*1, S Valenzuela2 techniques as a clear advantage over surgical venipuncture (i.e. femoral or jugular) and how it is often preferred to tail vein dosing as 1Animal Facility, Pontificia Universidad Católica de Chile, Santiago, the vein is superficial and can be visualized under the skin, enabling Chile; 2Fundación Ciencia para la Vida, Santiago, Chile more confidence in accuracy of IV dosing.

South American animal care and use programs are faced with the P62 Evaluation of Commercial Cling Film and Aluminum Foil for challenge of meeting international regulations and implementing the Use in Rodent Surgery necessary refinements to enhance animal welfare to raise standards. The region, however, is 10 years late in development compared to NA Celeste*1, K Emmer1, K Nolan1, T Collins1, D Mackessy1, MI America or Europe. To address international regulations and Perret-Gentil2, RA Malbrue1 recommendations, it was proposed to change from a caging system to a penning system for a new colony of New Zealand White rabbits. 1University Laboratory Animal Resources, Ohio State University, Staff assessed previously reported animal welfare improvements and Columbus, OH; 2Laboratory Animal Resources Center, The technical staff perceptions. This provided staff the opportunity to University of Texas at San Antonio, San Antonio, TX participate and contribute in this refinement process. Rabbits were reared in the same room, but were housed in a stainless steel wired- Aseptic technique, including the use of sterile drapes, is essential to floor caging system until weaning. To implement the housing system reduce microbial contamination to the lowest practical level and change, weaning-age rabbits from different litter mates are placed in improve surgical outcomes. Recently, some institutions have same-sex groups in the same pen. The pens were made in-house and approved the use of cling film (CF) as a practical, cost-effective were built to allow animals to exercise, play, forage, and show body alternative to sterile drapes for rodent surgeries. Additionally,

622 Abstracts of scientific papers 2019 AALAS National Meeting

aluminum foil (AF) has also been proposed to assist in maintaining a techniques on study. If multiple vet techs are available for a given sterile environment by covering surgical light handles, stereotaxic study, the technician with the most amount of practice takes apparatuses, surgical microscopes, and anesthesia machine knobs. precedence. This reward system was established to encourage the vet An additional benefit of CF relates to its insulating properties, which techs to practice and maintain unused skills. Over the last 4 mo, 100% provides an added layer of protection against hypothermia in the of our vet techs have demonstrated proficiency in these skills and are animal. The purpose of this study was to evaluate the sterility of CF able to assist investigators with these techniques in their ongoing and AF using ATP and RODAC plate testing. We tested 10 boxes of studies. This has also helped to create a closer bond between our CF and AF at days 0, 14, and 28 after opening the box and compared comparative medicine division and the research labs across campus. the results to traditional packaged sterile drape. Our data indicated that CF ATP bioluminescence remained at or below 10 RLU for 28 P65 Problems and Solutions in the Performing with CO2 as an dafter opening the box. Additionally, RODAC plates had no growth Euthanasia Agent in Mice for 70% of CF boxes at day 0, 100% at day 14, and 90% at day 28. The mean growth for the positive plates was 0.024 CFU/cm2 sampled PF Pohlig*1,2 after contacting locations on the front and back of the CF. Regarding AF, ATP bioluminescence remained at 0 RLU for 28 d after opening 1In Vivo Research Facility, CECAD, Cologne, Germany; 2Medres the box. RODAC plates had no growth for 80% of CF boxes at day 0, GmbH, Cologne, Germany 90% at day 14, and 100% at day 28. The results of this study support the use of CF and AF as acceptable alternatives for use during rodent Since the opening of a new large mouse facility in 2013 we have dealt aseptic surgery. intensively with the subject of euthanasia, as euthanasia was performed by both animal care staff and researchers. Use of carbon

P63 The Janet Wood Innovation Award: A Global Initiative to dioxide (CO2) or cervical dislocation are the main methods employed Invent and Introduce Novel Enrichment for euthanasia of mice. Even if trained in mouse euthanasia, people

often feel unsure how exactly to comply with CO2 euthanasia N Windows* procedures. However, wrongdoing or misuse is likely to cause immediate stress and pain for mice. First, we used a system to assure

Datesand Ltd, Manchester, United Kingdom a constant flow rate and slow increase of CO2 in purposely designed chambers. A remaining issue was that we could not guarantee the

Since it’s conception in 2016, the Janet Wood Innovation Award required increase of CO2 due to the design of the chamber. Therefore, (JWIA) has now launched more than 10 novel and unique products the procedure was purely visually monitored for signs of stress and into the global marketplace. The range of products includes a pain in mice. In order to to prevent possible misuse, we developed in number of items which allow mice to use the full 3-dimensional cage cooperation with a commercial company a fully automated fail-safe environment. There are also products which allow weaker or smaller solution that guarantees a constant and measurable increase of

mice to access the food hopper more easily and a solution to the CO2 (flowrate of 20% per minute), in which we use the air flow of the ongoing issue of wet, gel, or treat diet delivery. Every single one of existing IVC cage that fully complies with our and the requirements the JWIA winners has enriched the lives of our laboratory mice. of the AVMA Guidelines for the Euthanasia of Animals 2013. The

These products were designed by animal care technicians/animal concentration of CO2 increases slowly from 0.04% to 30% within the care personnel and veterinary technicians. From entering a first 3 min. At this point the anesthesia takes effect. After another competition with a concept product, the winners are selected by a minute the process is automatically stopped. panel of judges and voted on by their industry peers. The winning entries are taken from concept to marketplace with the help of P66 Dual IACUC and IBC Specialist Roles Allow for Cohesive corporate sponsoring team. Entering a design can be a daunting Approach to Protocol Review prospect, but we aims to dampen any fears and show how a sketched concept can be turned into a commercially available product which PP Gonzalez* can then have a positive impact on animal welfare around the world. Integrity, Oregon Health and Science University, Portland, OR P64 Development of a Training Program for Common Rodent Blood Collection and Dosing Techniques Comprehensive review of IACUC protocols requires input from multiple personnel from various departments across institutions. E Horrigan*, P Chamberlain, S Lackeyram-Owen, RM Kramer One of the major components of protocol review is to ensure consistency between IACUC protocols and work that requires Division of Comparative Medicine, Massachusetts Institute of approval from other committees. Frequently, researchers employ Technology, Cambridge, MA work that requires IBC review which may delay the IACUC protocol from being approved. While reviewer approval may be required Having a staff of veterinary technicians skilled in common rodent prior to the IACUC review proceeding, the IACUC administrator blood collection and dosing techniques is critical for research support reviewing the protocol may not have a complete understanding of and training of new personnel. Common techniques include work approved by other committees and thus work may be intraperitoneal injections, subcutaneousinjections, submandibular described incompletely. This may require multiple rounds of editing blood collection, and cardiac puncture blood collection. Since many and reviewing by both the researcher and the reviewers, both of researchers prefe rassistance with these procedures, our department which are time consuming. Furthermore, by having separate has developed a program to have all veterinary technicians trained reviewers oversee the review process of an IACUC protocol, it may on these techniques. Vet techs are trained by the training team, which result in a noncompliance issue if work is being done without consists of the vet tech supervisors and the training manager. approval from other regulatory committees. In order to streamline Training sessions are scheduled on an as needed basis as new staff the review process, a new position was created that combined the are hired. Once trained on these techniques, vet techs are required to IACUC and IBC offices to ensure that reviews of these protocols were practice on their own to become comfortable with performing the consistent with one another. This new position, research integrity procedure(s) on an active study. The training and practice sessions specialist, combines training from both the IACUC and IBC offices are tracked through a table that is readily accessible to the technicians and allows for reviews of both IACUC and IBC protocols, and the training team. Vet techs fill out each section, and then one of simultaneously if needed. This process alleviates the need for the the training team members approves the session. A minimum IACUC administrator to blindly hand off the IBC review to another number of practice sessions must be completed before the individual reviewer who may not know the requirements for the IACUC review is reassessed by the training team to evaluate proficiency. Once they and vice versa. Combining the reviews of IACUC and IBC protocols are deemed proficient, technicians are eligible to perform those allowed for both more complete and accurate reviews of both

623 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 protocols but also lead to consistency between both. We combine unreliability of serial measurements by traditional venipuncture. IACUC and IBC protocol reviews, thus allowing for better uniformity Patches remained adhered and functional on the pouched rats for an between protocols and to reduce review turnaround time. average of 10 d. Our data indicate that using a flash glucose monitoring system as a refinement for glucose monitoring is a P67 Behavioral Training to Increase the Welfare of Laboratory reliable way to obtain continuous glucose measurements Bovines noninvasively in this large rodent species, and has potential applications in other species. R Cohen*, C Chavez, G Blanco, A Kouneski, P Boliek, T Jones, M McDonald, H Jensen, R Gonzales, O Chiesa P69 The Evolution of Experimental Research in the Department of Medicine of the National University of Mexico Office of Research, FDA Center for Veterinary Medicine, Laurel, MD R Hernandez*1, r Guevara-Guzman2 Adverse behavior of bovines used in research can negatively impact the animal, study personnel, and the research study. We developed a 1Animal Facility-Unit, UNAM-Medicine School, Ciudad de Mexico, behavioral training program to decrease adverse bovine behavior, Mexico; 2Physiology, Medical School- UNAM, Ciudad Universitaria, increase bovine welfare and personnel safety, and prepare the cattle Mexico for standing minimally invasive surgical procedures. After training, the heifers will be able to walk calmly with a halter and lead rope We describe the evolution of experimental research in Mexico and its and stand calmly in the stanchion while restrained in a headgate. We relationship with medical training and education starting from the enrolled 32 Holstein heifers into the program in 4 cohorts of 8 heifers creation of the Clinical Science College (Colegio de Ciencia Medicas) each, with 16 selected for survey data collection. The heifers were in 1823 to the current Department of Medicine (DM) at the National trained to accept hand feeding, to allow study personnel to touch University of Mexico (UNAM). In the early 19th century, medical their heads, and stand calmly during restraint in the headgate. They training did not include research with animals in Mexico. Research in were next acclimatized to wear a halter, followed by walking on a animals was included in the syllabus when theories from the lead rope. The heifers were also trained to walk in and out of a positivism and Darwinism and cellular theory became relevant at the stanchion. The heifers were socialized by personnel up to 5 d per end of the century. In the veterinary medicine field, Dr Don Jose de la week for a minimum of 30 min per session. A qualitive and Luz Gomez was the pioneer in laboratory animal medicine in 1888. quantitative survey has been developed and will be used for the He was the first graduate from the earliest veterinary college at third and fourth cohorts. Prior to socialization, over 50% of the Mexico (1853). Early research with animals at the DM involved heifers would not initiate interaction with study personnel. As of researchers producing their own animals during the first half of the March 2019, 21 of 24 heifers in 3 cohorts have responded positively to 20th century. In 1984, the first central animal research facility of the staff, including initiating interaction, accepting hand feeding and UNAM was created and it houses the DM. For the last 35 years it has grooming, and voluntarily entering an open headgate. Heifers kept its name as Unidad Académica Bioterio (UAB) and continues to engaging in positive interactions with personnel were more provide support to the scientific community of the DM and other amenable to perform the desired behaviors for a reward. As of June research laboratories on campus. Although several natural disasters 2019 the last cohort have been enrolled in the socialization program, and disease outbreaks have occurred since its creation, the UAB has with most heifers initiating interaction with personnel. ByMarch successfully overcome these challenges and continued to enhance 2019, the program has successfully acclimatized 3 of the 4 cohorts of scientific research with animals at the DM-UNAM. The UAB was the heifers to personnel, wearing halters, and standing calmly in a first laboratory rodent breeding facility but the demand for stanchion and headgate. laboratory animals has exceeded the UAB capacities. Hence, this has lead to a complete facility renovation to improve our services and P68 Use of a Flash Glucose Monitoring System for Continuous laboratory animal care. Noninvasive Monitoring of Interstitial Glucose in a Southern Giant Pouched Rat (Cricetomys ansorgei) P70 Risk Compared with Harm-benefit Analysis: Its Impact on Evaluating Proposed Animal Studies by an IACUC or Comparable RN Labitt*, B Singh Oversight Body

Center for Animal Resources and Education, Cornell University, D Johnson2, RH Weichbrod*1, LJ DeTolla3, RC Simmonds5, P Ithaca, NY Houghton4, R Whitney6, L Kinter7,8

Monitoring blood glucose is a critical component of health 1Veterinary Research & Resources Section, National Institutes of monitoring for both clinical and research purposes. However, blood Health/National Eye Institute, Bethesda, MD; 2Cascades Biosciences collection requires restraint or anesthesia which can both artificially Consultants, Sisters, OR; 3School of Medicine University of induce hyperglycemia and increase stress to the animal. This can Maryland, Baltimore, MD; 4Primate Products Incorporated, make serial blood glucose monitoring impractical. A flash glucose Immokalee, FL; 5Laboratory Animal Medicine, University of Nevada, monitoring system in the form of a patch that measures interstitial Reno, NV; 6Retired Surgeon General of the United States, Steilacoom, glucose continuously for up to 14 d is available as an alternative to WA; 7GLP Scientific Consulting, Unionville, PA; 8Pharmacology & blood collection for diabetic human patients. This system has also Toxicology, Michigan State University, East Lansing, MI been validated in dogs. Here, we validate the use of the flash glucose monitoring system in the Southern Giant Pouched Rat, Cricetomys Currently, risk-benefits analysis (RBA) helps IACUC(s) or ansorgei. Blood collection in this species is impractical without comparable oversight bodies ensure that research with potential anesthesia, although small volumes may be obtained consciously by benefits using live animal models is of the highest quality, while venipuncture of the tail vein. We applied the patch to pouched rats addressing potential animal welfare concerns and managing a with and without spontaneously developed diabetes mellitus. stringent review/assessment method. This method helps ensure that Simultaneous readings taken using the flash glucose monitoring the animal(s) used in a study are not subjected to unnecessary pain, system, glucometer, and chemistry panels concurred. Oral glucose stress or distress, and/or injury. The process of protocol review for tolerance tests and insulin response tests were performed without potential adverse effects (animal welfare concerns) using RBA as requiring venipuncture and confirmed the clinical diagnoses of opposed to using a harm-benefit analysis (HBA) approach remains diabetes mellitus or normoglycemia. The continuous monitoring an important ethical consideration for organizations. Risk-benefit from the flash glucose monitoring system showed that the stress of analysis is a well-established method in animal research where the unanesthetized venipuncture elevated blood glucose by 25 mg/dl word “risk” provides for a prospective mitigation of potential which took approximately 90 min to return to baseline, indicating the adverse effects. In contrast, the use of harm-benefit analysis was

624 Abstracts of scientific papers 2019 AALAS National Meeting

recently introduced in the European Union (EU) using the word In theory, by addressing and correcting these weaknesses, learner “harm” as a replacement word for “risk” in the benefit analysis retention would increase and class engagement would be boosted, animal protocol review process. Words matter! Evaluating “risk” in and consistency amongst instructors would be improved. The the benefits analysis process is objective, unambiguous, and training team analyzed the workshop content and used historical quantifiable through clinical observation, experimental measurement, student evaluation data. As a result, 3 key topics emerged as the basis and probability analyses. Whereas using “harm” in the benefits for development of the new workshop. The class goal and learning analysis process is subjective, ambiguous, and emotionally charged. objectives were first established by the training team, and followed We postulate that harm-benefits analysis 1) contributes no advantage by development of 2-3 student learning activities per topic. The or improvement over the current risk-benefits analysis for the development of the learning activities solved the design flaws noted animals, investigators, or society; 2) miscommunicates to the public above. For example, the dense PowerPoint lecture was exchanged for by implying that animals are routinely harmed in research, thereby the activities that were received as engaging and enjoyable; damaging the research community’s reputation, the recruitment of additionally, the learning activities allowed a shift in emphasis from new scientists, technicians, and animal caregivers into the field; and trainer-centric content delivery to student-centric active learning. 3) increases the time and cost necessary to gain animal study Class evaluation surveys were provided to students at the end of approvals, delaying critical research, our increased understanding of each class with largely positive results indicating that students both biological processes, and the delivery of potential new treatments to enjoyed participating in the activities and left the class with a clearer improve the health and wellbeing of animals and people. understanding of the 3 key topics. Online post-tests sent out 1-2 mo from date of attending the class demonstrated that student retention P71 It’s Time to Rethink Critical Incident Reporting System in of the information had improved. Moving forward, this class will Laboratory Animal Science continually be analyzed to determine its effectiveness, with the intention to redesign areas of the class that remain problematic. SJ Bischoff*1, D Trietschel2, R Schiffner3, A Enkelmann2 Lastly, the successful aspects of this class redesign process can be applied for other classes. 1Animal Welfare, University Hospital Jena, Jena, Germany; 2Central Animal Facility, University Hospital Jena, Jena, P73 Characterization of Xenopus laevis Behavior and Enclosure Germany; 3Department of Orthopedics, University Hospital Jena, Space Use with Different Housing Densities in a Laboratory Waldkliniken Eisenberg, Eisenberg, Germany Environment

From today’s perspective and considering the current state of science, SP Kelly*, LL Diaz, RJ Tolwani use of animals in experimental purposes cannot be completely dispensed. Many scientific articles based on animal experimental Comparative Bioscience Center, The Rockefeller University, New studies are published daily. But negative experiences gained from York, NY these experiments get lost or are not referred in publications. The objective of the CIRS-LAS portal is the detection of critical incidents Appropriate housing of laboratory animals accounts for the physical, in the entire range of laboratory animal science. Thus, an open physiologic, and behavioral demands of the species by encouraging dialogue of failures can help to avoid them in the future. CIRS-LAS is naturalistic behaviors. Xenopus laevis is a commonly used laboratory the world’s first published web based critical incident database for frog species for which knowledge of optimal husbandry is limited, laboratory animal science and plays an exemplary pioneering role in with varied practices between institutions. For example, enclosure the implementation of the 3R principles. CIRS-LAS.de is based on space is allocated by many institutions as a volume of water per frog similar databases in human medicine and allows anonymous reports similar to other aquatic species. In the wild, Xenopus are found in of critical incidents. Currently, more than 90 people from Europe are shallow, stagnant bodies of water where they remain still at the water already registered users of the CIRS-LAS portal and more than 55 surface or floor, limiting movement to feeding or evading predators. critical incidents have already been entered. Registered persons can This has led some institutions to allocate floor space as opposed to comment and discuss the entered critical incidents to share their water volume for Xenopus housing, believing it allows for more experiences. In Europe, the scientific community fully agrees that naturalistic species behavior. We evaluated the current housing transparency and an open failure discussion are fundamental for standards by characterizing the behavior and enclosure space use excellent animal-based research. After a widely spread acceptance of Xenopus laevis. Groups of adult, captive-bred Xenopus laevis were among Europe we would be pleased to introduce CIRS-LAS.de to the housed at 5 frogs per enclosure and provided with 25.2 liters of water U.S. The collaboration of our approaches in work according to the 3R per frog. The groups were recorded for 5 d and their behavior was principles will enhance animal welfare worldwide. We are convinced evaluated at defined intervals to characterize: time at motion versus that the implementation of CIRS-LAS serves to enhance the trust in time at rest, location at rest, and interaction with conspecifics. The laboratory animal science of both the public and scientific same conditions were then characterized for 1 group by increasing community. The time to rethink has been achieved: to learn from the stocking density (16, 32, and 63 frogs per enclosure) over time, negative results in animal based research. thus decreasing the amount of available space per frog (8, 4, and 2 liters per frog, respectively). It was found that, on P72 A Structured Approach For Revamping Institutional average, Xenopus remained at rest over 69% for the time reviewed at Workshops all housing densities, the majority of which occurred on the enclosure

* floor. SY Nowlan , M Kellepourey, KA Pokryfke, MC Dyson P74 Evolving the Role of Discovery-focused Pathologists and Unit for Laboratory Animal Medicine, University of Michigan, Ann Comparative Scientists in the Pharmaceutical Industry Arbor, MI S Mohanan, S Maguire*, J Klapwijk, R Adler, P Clements, R Haworth The animal program provides a 2-part training workshop entitled “Animal Room Procedures for Rodents” that instructs (part 1, online) TMCP, Glaxosmithkline, Collegeville, PA and reinforces (part 2, instructor-led) the expected practices for working in a rodent housing room. As part of a regular evaluation of A pharmaceutical company has recently made significant class offerings, the training team determined that part 2 had several organizational changes to its nonclinical safety, drug metabolism and design flaws in its original form, including a lengthy PowerPoint pharmacokinetic, and laboratory animal science/veterinary handout for students that included too much jargon, too many functions, with the goal to increase our focus on scientific partnership different class topics, few active learning opportunities, and ample with the discovery part of the organization. One specific change was opportunities for various instructors to teach the material differently. bringing together pathologists and comparative medicine

625 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 veterinarians and scientists into a single functional unit. We describe sessions, sometimes up to 5 per week. Furthermore, the lecture our early activities (assessing our capabilities and gaps, external format consisted of reviewing a handout, which did not engage lab benchmarking, listening to our discovery partners, redesigning some personnel’s attention. In 2017, orientation training was converted to of our working practices) aimed at implementing these changes. In an online course using learning management software provided by addition, early on we held a discovery engagement workshop the institution in an effort to alleviate some of these issues. Online attended by all pathologists and comparative medicine veterinarians training is becoming an essential learning tool for laboratory animal and scientists, as well as selected discovery scientists. The purpose of science professionals and a survey conducted by our institution’s this workshop was to share learnings from the above activities and IACUC indicated that 50% of lab members would prefer web site devise plans aimed at achieving our overall goal of functional material and online courses. The benefits of online training include integration: driving pathobiology expertise into drug discovery and the ability to incorporate knowledge check questions to confirm that increasing the human (translational) relevance of experimental data. key points are retained; the ability of lab members to complete We describe the new organizational structure, the workshop training on their own time and at their own pace; language activities, and implementation plans; updates on our progress; and translation and closed captioning options to meet the diverse needs considerations for a pan-industry network of discovery-focused of laboratory animal science personnel; and the ability of lab pathologists and comparative medicine veterinarians and scientists. members to return to the site to reference online training material. Our records indicate that 64% of those who complete our training P75 An IQ Consortium Perspective on the Scientific Committee on return to reference the material. Since implementing online Health, Environmental, and Emerging Risks Final Opinion on the Orientation, our online training library has expanded to include Need for Nonhuman Primates in Biomedical Research, Production, nonhuman primate training. Due to the success of online orientation and Testing of Products and Devices training, additional online courses are being developed for rodent breeding and colony management and pain and distress in S Maguire*2, KA Adams1, D Clemmons3, L Collura Impelluso4, D laboratory animals. Lee5, A Myers6, C Petursson7, R Schulingkamp8, K Trouba8, M Wright9 P77 MR and CT Compatible Electrical Heating System for Mouse Imaging 1Incyte Research Institute, Wilmington, DE; 2Integrated Biological Platform Sciences, GSK, Collegeville, PA; 3Comparative Medicine, S Gilchrist1, V Kersemans1, PD Allen1, AL Gomes2, S Wallington*1, P AbbVie, North Chicago, IL; 4Comparative Medicine, Pfizer Kinchesh1, SC Smart1 Worldwide Research and Development,, Groton, CT; 5Department of Safety Assessment, Genentech,, South San Francisco, CA; 6IQ 1Oxford Institute for Radiation Oncology, University of Oxford, Consortium, Washington, ; 7Department of Veterinary Sciences, Oxford, United Kingdom; 2The Francis Crick Institute, London, Bristol-Myers Squibb, Princeton, NJ; 8Janssen Research & United Kingdom Development, Spring House, PA; 9Department of Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, CA Anesthesia induces a rapid onset of profound hypothermia with its physiological complications, but normothermia can be maintained The recent Scientific Committee on Health, Environmental, and through careful, active heating. It is common to warm anesthetised Emerging Risks Final Opinion on the need for nonhuman primates in animals using electric blankets, heat lamps, warm air, or water baths. biomedical research, production, and testing of products and devices Most commercial systems for heating animals within tomographic highlights approaches that could significantly contribute to the (3D) imaging systems use circulating warm air or warm liquid, both replacement, reduction, and refinement of nonhuman primate (NHP) of which require free space around the animal. Where space is studies. Initiatives that have the potential to affect NHP welfare and/ limited small resistive heater elements are effective. These use copper or their use are expected to be appropriate, fair and objective, and wire heating elements and are MR-compatible; images are not publicly disseminated information focused on NHPs in biomedical distorted through their use. For x-ray CT imaging, however, the research, which includes toxicologic and pathologic research and copper distorts the images significantly due to interactions between testing, should be objectively evaluated by stakeholder scientists, the x-rays and the copper atoms, an effect not seen for smaller researchers, and veterinarians. Thus, IQ Consortium member elements in periods 1 and 2 of the periodic table (hydrogen-neon). Of companies convened to develop an informed and objective response, the electrically conductive smaller elements (lithium, beryllium, and focusing on identifying areas of agreement, potential gaps or missing carbon) only carbon is chemically stable and nontoxic. Carbon is information. Overall, the authors agree that many positions in the widely used as an electrical conductor and we have developed an opinion generally align with industry views on the use of NHPs in electrically powered resistive heater element using carbon fibre research and testing, including the ongoing need of NHPs in many sheeting that is both MRI and CT compatible, and which also works areas of research. From the perspective of the IQ Consortium, there effectively on the bench. The heater elements are also cost effective to are several topics in that merit additional comment, attention or produce. CT compatibility results from use of carbon; MR research, as well as consideration in future opinions. compatibility through the use of 100 kHz alternating current. The heater is integrated in a control loop such that animals are warmed to P76 Incorporating Online Training into an Animal Care Program a target temperature, but through operation at a fixed maximum temperature, the heater elements are incapable of causing direct S May* thermal burns consequent to being driven at full power as can happen with some commercial heaters. We demonstrate the efficacy Research Animal Resources, University of Minnesota, Minneapolis, of the technique using MRI, CT, and PET and SPECT images. The MN latter requires high-fidelity CT images in order to allow quantitative analysis of the image intensities such that quantitative tracer uptake In accordance with the Guide’s requirement of adequately training analysis is possible. By implementing this system inhouse, we can personnel in basic principles of laboratory animal science and animal deliver fast and accurate multimodal imaging where the animals’ wellbeing, our facility requires new lab personnel to complete physiology is balanced and maintained allowing shortened orientation training before working with animals in the vivarium. anaesthetic duration and faster recovery times with no welfare The main topics covered in orientation include facility guidelines, complications. animal acquisition, husbandry, and health monitoring. The previous format of orientation was a 1-h classroom lecture offered twice a week. This was often inconvenient for lab personnel and facility staff. The recurring weekly schedule frequently conflicted with lab personnel availability and would require scheduling additional

626 Abstracts of scientific papers 2019 AALAS National Meeting

P78 Going against the Grain: Starting a Postapproval Monitoring P80 Refinement of Core Body Temperature Collection in Program under the Attending Veterinarian Cynomolgus Macaques (Macaca fascicularis) Using Implantable Temperature Microchips S Patton*, P Charlton S Deans* Division of Laboratory Animal Services, Augusta University, Augusta, GA Training & Compliance, Envigo, Long Valley, NJ

As our research program grows, the supporting departments such as Core body temperatures give invaluable insight to physiological the Division of Laboratory Animal Services (DLAS) have had to changes and general animal health and as such are a critical make many changes to keep pace. Following our last AAALAC site component in pharmacokinetic and biologic drug analysis. visit findings, our Animal Leadership Committee (ALC, comprised of Historically, rectal probes have been used to determine core body the Senior Vice President of Research, the Associate Vice President of temperature and are proven to provide reliable and accurate data. Basic Research, the attending veterinarian, the DLAS Director for This method is an invasive procedure that requires considerable Husbandry Services, and the IACUC Chair) determined a need to handling and restraint, as well as safety considerations. To refine the create an official postapproval monitoring (PAM) program to core body temperature procedure in nonhuman primates, improve promote alignment between laboratory practices and approved the wellbeing of our animals, and minimize safety issues, this trial animal use protocols. Since launching the program in January 2018, evaluated implanted microchips to collect core body temperatures we have faced many unique challenges, particularly in regard to how with a high-powered reader, without removing the animal from its the position interfaces with the IACUC (since the PAM program is cage. Four cynomolgus macaques (Macaca fascicularis) were not part of the IACUC office) as well as researcher perception of the implanted with 2 identical microchips each, 1 in the intraperitoneal PAM program. Now that we are over 1 y into the new program, we cavity, the other in the scapular region (implanted subcutaneously) have developed an effective reporting structure with the IACUC to Animals that had been previously implanted with a telemetry device discuss and relay findings and ensure closure of documentation. We were selected due to the additional body temperature reading have also developed a collegial rapport with the research staff, as comparison. Each method of collection was compared to a rectal demonstrated by the rate of compliance for amendment submission temperature to determine reliability and accuracy. Temperatures following initial PAM notification. were recorded immediately after implantation and for 4 consecutive d thereafter. The collections were repeated once weekly for a further P79 Refinement of Heat Sources to Support Animals Exposed to 4 wk to assess the proper functioning of the microchips and readers. Accidental Cage Flooding After completion of the trial, it was shown that the intraperitoneal implanted microchip gave reliable data, averaging within 0.9% of the S Pittsley*1, FC Hankenson1,2 rectal temperature values. The telemetry readings were within 1.7% and the subcutaneous values were within 2.7% of the rectal readings. 1Campus Animal Resources, Michigan State University, Lansing, When data was collected with the microchip using the high-powered MI; 2Department of Pathobiology and Diagnostic Investigation, reader, technicians were able to identify each animal and collect the College of Veterinary Medicine, Michigan State University, East temperature data by simply moving each animal to the front of the Lansing, MI cage. This reduced handling stress and exertion for each animal involved enabled technicians to get data safely and at multiple time An infrequent, yet serious, husbandry situation that requires intervals. The intraperitoneal implant readings consistently remained emergency intervention is the flooding of rodent cages following an close to the rectal temperature values throughout the trial and were equipment malfunction. Water-soaked bedding, after release of a full considered to be a reliable and accurate replacement for rectal bottle to simulate a flood, registered by temperature probe at a low of temperatures. 20°C (~67°F), which is ~30% below the rodent preferred thermoneutral temperature of ~28°C, and which can lead to P81 The Role of Preclinical Rodent Services in Academia: The Why irreversible hypothermia and mortality, particularly in altricial pups. and How In our current practice, a flooded cage is promptly reported to the veterinary staff, animals are dried off by care staff and placed into a TA Cox*, R Bolin, NA Zielinski-Mozny clean bedded cage. Depending upon supplies available in any given animal facility, heat lamps, heat discs, or warm-water recirculating Center for Comparative Medicine, Northwestern University, blankets have been used to provide thermal support. Due to the Matteson, IL variability of the approach to rewarming rodents, we sought to identify a reliable and economical heat source to rewarm animals, Researchers spend a significant amount of time designing while also creating an SOP for staff to consistently use across all experiments, performing in vitro bench work, writing papers, and rodent facilities. After assessment of numerous devices, we found completing grant submissions. These activities are time consuming that a commercial terrarium heat mat in combination with a 75-watt and compete with time for training on rodent technical procedures heat lamp clipped beside the cage were able to raise the bedded cage that are required to complete experiments. The concept of a Pre- temperature ~9°C within 10 min, to ~30°C, regardless of bedding Clinical Rodent Services (PCS) was initiated based on researcher type. Temperatures within the clean cages continued to rise over requests for training on these techniques which may take a time; within 20 min they were heated to 34°C (paper chip), 34.5°C considerable amount of time to master. For example, laboratory (wood chip) and 32.4°C (corncob). Based upon these data, we have members who are not proficient with the tail vein injection technique created an SOP to direct staff to clip the lamp bulb at 6 in above the would not have to go through training to have this procedure cage level and place the terrarium mat halfway under a clean bedded performed as part of an experiment. Thus, the PCS was created to cage to allow for a temperature gradient. Animals will be kept provide researchers with a pair of in vivo technical hands. The exposed to the heat lamp for 10 min, at which time the lamp will be service was generated from basic techniques that researchers needed turned off, and the mat will remain in place for as long as vet staff for drug discovery activities and management of breeding services. recommend. Flooded cage recovery ‘kits’ with pictures of how to The use of the PCS has increased over time with researchers asking set-up all needed equipment will be available in each facility (n=10), for technical assistance with a variety of techniques approved in their with the expense being ~$43 per kit. This data-based approach to protocols. This includes oral gavage, tail vein or retroorbital address a spontaneous welfare issue in laboratory rodents will injections, blood collection, and colony management (i.e., tail biopsy, provide confidence to our department to best support the return of ear tagging and weaning). This service can also potentially save the body temperatures and activity levels in our rodent patients that principle investigator funds, as they may not need to bring on an have experienced a flooded cage. additional team member to perform such tasks. The PCS technicians

627 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 are readily available to assist with specified tasks and help reduce the role and contribution for all committee members to understand. load of responsibilities on each researcher in a time efficient manner. Training for our online protocol software has also been implemented. The PCS provides the animal biomedical research community with We created an onboarding member survey to provide to new high-quality, technical expertise to support research projects. With committee members after training to evaluate our new process. For increased advertising (word of mouth, flyers, poster sessions) over this project, we believe the changes we have made not only the past 6 mo, PCS has become more visible to and better used by streamline the onboarding process, but also improve the material researchers. provided. Our ability to truly address the success of our onboarding program will be slowly gathered as we train new committee P82 Reduced Sample Volume for Increased Animal Benefit: Using members. In conclusion we believe we have restructured our IACUC Microsampling and Low-volume Blood Collection Techniques for member onboarding to provide a better education process for all Toxicology Studies members of the committee. As we onboard new committee members the survey we created will help determine if our new process is T Putnam*, C Foppiano successful. It is important for institutions to continually evaluate their training program to ensure the IACUC committee can Toxicology Study Operations, Boehringer Ingelheim, Ridgefield, CT effectively evaluate protocols and animal welfare issues.

When conducting nonclinical toxicology safety studies, blood volume P84 Programmatic Evaluation of an Institutional Animal User availability (per IACUC guidelines) is commonly a limiting factor, Training Program and Its Role in Ensuring Trainee Success which can restrict the number of endpoints supported, and is often accounted for by increasing the total number of animals on study or V Oliver*, M Gillespie, M Trotter, C Geary Joo, K Hecker, D Morck adding satellite groups. As such, there is a constant desire to decrease the overall blood volumes collected from animals, while still University of Calgary, Calgary, Canada generating high-quality study data. Supporting increased sample requests on studies (both GLP and nonGLP) with decreased sample Institutional animal user training programs (IAUTP) are essential volumes required the development of innovative blood collection components of research organizations ensuring appropriate training processes. Refined techniques were needed that would consistently of personnel and satisfaction of mandatory regulatory requirements. yield quality samples, with minimal to no blood loss, and could be Evaluation of these programs is critical to confirming effectiveness of applied across the large and small animal species used for toxicology personnel to uphold high standards of ethical and humane research studies. To address this need, a conscious jugular ‘micro-sample’ with animals, but is often overlooked. We describe evaluation of our collection technique (50µl) for rats, and a ‘low volume’ cephalic/ IAUTP after programmatic redesign using Kirkpatrick’s 4-level saphenous collection technique (600µl) for large animals (beagle/ evaluation model. Self-assessed confidence and satisfaction were cyno) were developed, and found to have a direct benefit for serial evaluated to gauge trainee reaction. Trainee success in theoretical and collection of toxicokinetic (TK) samples. These techniques enabled behavioral-based assessments were measured to assess learning and the collection of TK samples using minimal sample volumes, and behaviour. Within the redesigned program, trainee confidence maintained the same (or more) time points, while significantly significantly (P < 0.002, D > 1.0) increased in post-core mandatory decreasing the overall volumes collected for each animal. Reduced content confidence scores, and continued to significantly increase overall collection volumes allowed for additional sample type (P < 0.001, D > 0.7) as trainees progressed throughout the program. collection, of particular value in primates and rats. These techniques Trainee satisfaction was higher (P < 0.001, D > 0.6) in the new have been successfully used on numerous nonGLP and GLP studies, program for the areas of program delivery and engagement. in all 3 species, and on both oral gavage and IV dose toxicology Evaluation of trainees in both theoretical (45.3 + 2.8 out of 50) and studies. The decreased volume collection approach has also been performance-based (average 3.7 + 0.3 out of 5) assessments were high applied to alternate sample types, including antidrug antibody in the new program. Overall, the new training program was well (ADA) and metabolite identification (MetID), further decreasing the received and appears to be producing successful trainees. Further overall blood volume collected on study. Implementing microsample research to measure trainee competence and performance outside of and low-volume collection approaches allowed for expanded an instructional environment are recommended to improve future individual animal endpoint sampling, opportunity for reduction of data collection, as well as the establishment of additional metrics to animals (e.g., elimination of satellite TK groups), and ultimately more accurately measure higher levels of programmatic evaluation. improved animal welfare in support of the 3Rs. P85 Duration of Patency of Jugular Vein Catheters Attached to P83 Onboarding of New IACUC Committee Members Transcutaneous Buttons in CD1 Mice without Maintenance

TM Hadley*, A Lantron V Karicheti*, A Kang, C Ortiz, M Koubaitary, EB Ashford, DL Denton, Y Luo, A Williams IACUC, University of Colorado Boulder, Boulder, CO Surgery, Charles River Laboratories, Raleigh, NC The IACUC committee is a vital part of animal research and ensures the safety of the animals. It is important that members of the Central venous access in conscious mice for repeated blood sampling committee have proper training in order to review protocols and in pharmacokinetic studies is achieved using chronically implanted make decisions to the best of their abilities. During an internal review jugular vein catheters (JVCs). However, maintaining continuous of our program we felt that our training program needed patency sets practical limits on its uses. We have previously shown improvements. Our goal was to provide sufficient training for that the use of transcutaneous buttons kept the catheters patent members to help them use their area of expertise while serving on longer than the standard externalized catheters. One factor that the committee. Our aim was to provide appropriate training without affects catheter patency is the catheter maintenance schedule. We overloading new members. In order to improve our onboarding conducted a study to determine the duration of blood collection system, we reached out to other institutions to see what they found patency and infusion only patency of JVCs attached to successful in their onboarding processes, and interviewed a few of transcutaneous buttons in mice without catheter maintenance. Fifty our current members, including the nonscientist and unaffiliated adult male CD-1 mice at 37-39g (60-63-d-old) were randomly members, to understand how they felt their training could have been assigned to 5 groups (n=10 each). Mice were anesthetized and a improved. Once we collected the responses we began restructuring polyurethane catheter was inserted into the right jugular vein. The our training. One area we wanted to focus on was the training for the catheter was connected to a transcutaneous button in the nonscientist and unaffiliated members. To do so, we now provide interscapular region. The catheter and transcutaneous button were additional information for those members and also emphasize their locked using heparinized (500 IU/ml) 50% dextrose solution. Patency

628 Abstracts of scientific papers 2019 AALAS National Meeting

was checked for each group at 1, 2, 3, 4, and 5-wk postsurgery working principles and the role of IACUCs at the CDC and Emory without any other manipulation. Animals were manually restrained, University have been reviewed and summarized. Considering and catheters accessed using a button adaptor injector. The catheter various opinions about adaptation and implementation of was aspirated to confirm the ability to withdraw blood. If aspiration international standards for biomedical research in Georgia and based failed, a saline flush was attempted. If the flush solution freely on objective reality, it was concluded as appropriate for the first stage infused, a second aspiration was attempted. The catheter was to establish the Interinstitutional Animal Care and Use Committee at considered fully patent if withdrawal of blood was successful with National Center for Disease Control and Public Health (NCDC) of the first or second attempt. Animals were clinically healthy Georgia. As a result, in 2019 the committee was established that now throughout the study. Catheter patency was 100% in the wk 1 to wk 3 allows to conduct and monitor all ABSL-2/ABSL-3 studies in groups postsurgery. Patency rates decreased to 90% and 60% at wk 4 accordance with international standards. As well, memorandums of and wk 5, respectively. None of the catheters were blocked and collaboration were signed between GALAS and major Georgian successful infusion was maintained through wk 5 after surgery. This biomedical institutions, including NCDC, Center for Experimental data suggests that bidirectional catheter patency for blood collection Biomedicine, and Tbilisi State University. GALAS provides all the is relatively shortterm in comparison to catheter patency for infusion necessary documentation to above mentioned institutions and assists only of mice JVCs attached to transcutaneous buttons in mice in the proper preparation of IACUC staff to increase their without catheter maintenance. competence consistent with the international standards and regulations. Thus, the first important milestone in the development P86 Mission for Ethical Animal Experiments Education Program at of sustainable IACUCs system has been successfully achieved. Seoul National University Hospital P87b Strategy for the Implementation of Biomedical Research Legislation in Countries in Transition: Georgian Experience Y Won *, J Parl, B Kang L Chitiashvili*1,2, K Mulkijanyan1,3, M Ramishvili1,2, P Imnadze2 Seoul National University Hospital, Seoul, Korea 1Georgian Association for Laboratory Animal Science, Tbilisi, In Korea, the pet industry and social ethics awareness is developing Georgia; 2National Center for Disease Control and Public Health, rapidly, and the interest and debate on animal experiments and Tbilisi, Georgia; 3Tbilisi State Medical University I. Kutateladze protection are also increasing, Our department has been supporting Institute of Pharmacochemistry, Tbilisi, Georgia various preclinical studies for medical research since 1998. We were certified as KGLP facility for safety study of new drug and medical During the past decade, Georgia has fostered collaboration with devices in 2003, and accredited by AAALAC International in 2007. scientific and educational organizations both in the US and EU. In During the last 20 years, we have been operating several education turn, this process requires significant changes in regulation of programs for the staffs and investigators in animal research facility biomedical research. State-of–the-art requirements for biomedical based on the 3Rs principle in order to keep with the social needs and studies suggest, on one hand, proper regulations and guidelines for the global trends of ethical animal experiment. Before starting animal use of laboratory animals involved in the research process, and on experiments, one should complete 2 mandatory educations (1) a the other, availability of relevant vivarium infrastructure and monthly animal experiment technique hands-on workshop and 2) qualified staff. Formerly, the Georgian research institutions each and a biweekly new user guidance education for animal research carried out experimental research based on their own considerations, facility. The researcher who is conducting animal experiments should as no policy on use and care of laboratory animals existed, and this participate in the refresher training program as well as an ethical tradition is partially still in force. However, nowadays the absence of animal experiment seminar bimonthly. For the staff in animal legislative basis along with mostly amortized animal facilities and facilities, weekly staff training program is conducted with contents equipment has negative impact on participation in international based on each staff’s job description and an annual team building scientific projects, obtaining funding for the research, as well as workshop is held to improve the management condition and set up publishing in peer-reviewed journals, due to noncompliance with the future plans. We educate approximately 450 new investigators internationally accepted standards and hence lack of credibility of every year, and do the best efforts to raise the level of ethical results. Since 2016 the Georgian Association for Laboratory Animal consciousness and work ability of experimental animals. We set the Science (GALAS) aims to contribute to biomedical research by step- 2019 goals on proper housing, high quality, good support, and a safe by-step implementation of laboratory animal care and use policy in work environment. We expect that these educational programs can Georgia. As an internationally recognized organization, a scientific lead to the development of medical research and animal welfere in member of ICLAS since 2016 and observer at FELASA since 2018, Korea. GALAS provides interested parties with all the necessary information regarding contemporary ethical standards and regulations in animal P87 Building of IACUCs System in Georgia: Story of Success housing, nutrition, and routine operations. Georgian universities and research institutes are now provided with the unified set of M Ramishvili*2,1, K Mulkijanyan2,3, L Chitiashvili2,1, P Imnadze1 regulatory documents. However the ratification of the European Convention for the Protection of Vertebrate Animals used for 1National Center for Disease Control and Public Health, Tbilisi, experimental and other scientific purposes, along with the phased Georgia; 2Georgian Association for Laboratory Animal Science, implementation of the fundamental provisions of EU Directive Tbilisi, Georgia; 3Tbilisi State Medical University I. Kutateladze 2010/63/, remain among the main objectives for further success and Institute of Pharmacochemistry, Tbilisi, Georgia recognition of Georgian biomedical science. Currently, GALAS is developing a draft of appropriate amendments to Georgian The absence of national legislation regarding experimental animals legislation for submission to the Parliamentary Committee on Science negatively impacts both the credibility of Georgian scientific and Education. products, regardless of their scientific value and international collaboration. Local scientists are forced to be temporarily guided by P87c The Application of Clinical Scoring Systems in Foreign international regulations which can be challenging. Significant Animal Disease Research changes in the management of biomedical and laboratory animal science are urgent to fill this gap. Among these, the development of J Hagert* IACUCs is an important step to national policy on care and use of experimental animals. The Georgian Association for Laboratory Plum Island Animal Disease Center, Old Saybrook, CT Animal Science (GALAS) carried out several workshops devoted to the establishment of IACUCs in Georgia. The AALAS and FELASA We developed a clinical scoring system specific to each foreign qualification criteria for personnel working with animals and the animal disease we study to meet AALAS and IACUC standards to

629 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 minimize pain and distress in animals. The system also provides staff intracorneal fungal organisms consistent with Batrachochytrium with veterinary care guidelines, which can help prevent compassion dendrobatidis, as well as emaciation and nematodiasis of various fatigue that may occur while working in an animal research facility. organs. Following the diagnosis, the final 3 surviving frogs in the Veterinary experts, experienced in the progression of each foreign colony were euthanized, with skin swabs from all 3 submitted for animal disease, developed the system, specific for each disease, to molecular diagnostics and 2 frogs submitted for necropsy. Of these, 1 allow staff to recognize clinical signs that they may encounter frog was positive for chytrid fungus by polymerase chain reaction throughout the day. The numerical scale ranges from 0 to 5, and histopathology. All 3 frogs were negative for ranavirus. The 6 categorized by severity of clinical parameters. A score of 0 reflects an remaining frogs that were not submitted for diagnosis were found animal showing no clinical signs while a score of 5 signifies an dead. The colony collapse was attributed to chytridiomycosis, with animal in a moribund condition. Any animal with a clinical score of 4 emaciation and endoparasitism as concurrent factors. Investigations or 5 is reported to the veterinary staff for immediate welfare into facility and water improvements are ongoing, with expected evaluation and a treatment plan that provides compassionate care, aggressive decontamination and quarantine procedures before alleviates pain and distress, or results in euthanasia when the use of repopulation. analgesics or other therapeutics cannot improve or assuage conditions that are predictive of impending death. Increased P90 Endemic Pneumocysits spp Infection Resulting in Morbidity awareness of these predictive signs through clinical scoring has and Mortality in Immunocompetent Mouse Colony improved the prevention of spontaneous deaths among research animals. Application and success of the clinical scoring system relies AL Armijo*1,2, Z Ge1, N Fabian1, D Puglisi1, JM Essigmann2, RG on a combination of expertise in a research facility. Trained animal Croy2, JG Fox1, S Carrasco1 care staff are essential in providing accurate clinical assessments throughout the day and effectively communicating these with 1Division of Comparative Medicine, Massachusetts Institute of veterinarians and investigators who design clinical care plans. Technology, Cambridge, MA; 2Departments of Biological Engineering Animal care staff experienced with the clinical progression of each and Chemistry, Center for Environmental Health Sciences, foreign animal disease provide excellent, humane care of the research Massachusetts Institute of Technology, Cambridge, MA animals resulting in high-value, reliable clinical data, which are used yearly in at least 20 peer-reviewed publications from our institution. A 2.5-mo-old, 20.9 g male C57Bl/6-TgN(gpt Delta) used in a chronic ethanol treatment study (9 d on a Lieber-DeCarli liquid diet P88 Determination of Vertebral Heart Score Standards in Owl containing ethanol) presented with a hunched posture and a greater Monkeys (Aotus spp) than 20% loss of body weight. Physical exam revealed severe dehydration. All other animals on the study, including controls, were A Brown*, ML Thomas, K Cogan, L Colenda, B Cordero mildly dehydrated, but otherwise clinically normal. Water bottles were provided in addition to the liquid diet, but 4 animals in the DVR Surgical Service, National Institutes of Health, Bethesda, MD treatment group were found dead 2 d later. The study was terminated and the remaining animals were necropsied. Postmortem Heart disease is prevalent in owl monkeys (Aotus spp) used for examination indicated that the lungs of mice were diffusely red to biomedical research. Because of this tendency for heart disease, we pale-pink, firm, and failed to collapse. Histologically, pulmonary wanted to develop a screening tool, usiing the Vertebral Heart Score lesions in all of these mice (15/15) were characterized by histiocytic (VHS) to provide a baseline measurement on newly acquired and neutrophilic interstitial pneumonia with alveolar histiocytosis primates to look for early signs of cardiomyopathy and compare it to and pulmonary edema. A subset of these mice (10/15) exhibited scores obtained later in life. The VHS is used by veterinarians to intrahistiocytic and extracellular cysts and trophozoites in alveolar evaluate the heart and determine the presence or absence of spaces. The presence of yeast forms was confirmed by Gomori cardiomegaly. By measuring the long and short axis of the heart and methenamine silver and Periodic acid–Schiff staining. One of 15 mice then using the mid-thoracic vertebrae as a unit of measurement, the had concurrent eosinophilic crystalline pneumonia, and 9 of 15 mice heart size can be determined. The aim of this study is to determine had pulmonary arterial medial hypertrophy with mild neutrophilic Vertebral Heart Score Standards for aotus monkeys. Vertebral Heart arteritis. Pneumocystis DNA was amplified by PCR from the lungs of Scores were calculated from right lateral thoracic radiographs on 150 10/15 mice. Amplified PCR products from formalin-fixed paraffin- owl monkeys. Results of our survey determined that the normal VHS embedded lung sections were obtained from the 18S rRNA gene. The range for Aotus monkeys is 9.0–10.7. This noninvasive radiographic 18S rRNA sequences derived from these cases shared 99-100% technique will benefit nonhuman primate veterinarians in objectively identity with P. murina DNA. Aerobic cultures from lung sections of evaluating cardiac size and cardiovascular disease in aotus monkeys. these animals were negative for primary bacterial pathogens. Since Pneumocystis DNA has been confirmed in this colony, the lungs P89 Cope’s Gray Treefrog (Hyla chrysoscelis) Colony Attrition and from 5 naïve mice and 7 naïve mice with clinical signs of illness, Chytridiomycosis including increased respiratory effort, have tested positive for Pneumocystisby PCR. Our findings confirmed AM Edmunson*1, TW Carlson2, JA Dykstra2, FD Duke Boynton1 endemic Pneumocystis infection with associated morbidity and mortality in this colony of immunocompetent mice. The colony has 1Research Animal Resources, University of Minnesota, Minneapolis, been rederived through embryo transfer and additional preventative MN; 2Department of Veterinary Population Medicine, University of measures, including depopulation of the infected colony. Minnesota College of Veterinary Medicine, Saint Paul, MN P91 Vertebral Heart Score in Rhesus Macaques (Macaca mulatta) A colony of Cope’s gray treefrogs (Hyla chrysoscelis) presented in with and without a Cardiomyopathy and Establishing Reference October 2018 for severe attrition of unknown etiology. The colony Intervals consisted of wild-caught frogs used for auditory research, and were group-housed in modified flow-through aquatic tanks. Over 100 AR Williams*3, Y Ueda2,3, K Christe3, JA Stern1,3 frogs were moved to a new location in April 2018, with loss of 50% of the frogs each month thereafter, resulting in a colony size of 10 frogs 1Cardiology, UC Davis-Veterinary Medical Teaching Hospital, Davis, at presentation. The affected frogs were found dead with no previous CA; 2Emergency and Critical Care, UC Davis-Veterinary Medical clinical signs. Water quality parameters evaluated by a third party Teaching Hospital, Davis, CA; 3Primate Medicine, UC Davis- laboratory were determined to be within acceptable ranges. The only California National Primate Research Center, Davis, CA clinical sign observed on colony exam was skin discoloration. One animal was euthanized for necropsy examination; findings included Vertebral heart score (VHS) is an established screening tool to epidermal hyperplasia and hyperkeratosis with numerous determine the presence of cardiomegaly in humans and small

630 Abstracts of scientific papers 2019 AALAS National Meeting

animals. VHS provides a semiquantitative assessment of splenomegaly in immunocompromised mice. Careful monitoring of cardiomyopathies based on thoracic radiographs and its use is sterilization techniques is essential to prevent such infections in a largely lacking in primate medicine. Here, several rhesus macaques barrier facility. have been diagnosed with various cardiomyopathies by echocardiographic examination. We hypothesized that rhesus P93 Vascular Hamartoma in the Uterus of a New Zealand White macaques with cardiomyopathies have a higher VHS than those Rabbit without cardiomyopathies and VHS could replace echocardiography as a screening tool. In this study, VHS was measured for 126 rhesus A Merley*1, JA Dykstra2, M Dunbar3, J Hubbard1 macaques using the right lateral (RL), dorsoventral (DV), and ventrodorsal (VD) views of thoracic radiographs. Of the animals 1Research Animal Resources, University of Minnesota, Minneapolis, evaluated, 37 were diagnosed with various cardiomyopathies by MN; 2Veterinary Diagnostic Laboratory, University of Minnesota, St. echocardiographic examination: 31 with hypertrophic Paul, MN; 3The University of Texas Southwestern Medical Center, cardiomyopathy (HCM) and 6 with nonHCM cardiomyopathies. Dallas, TX Reference intervals were established in the rhesus macaque without any identifiable cardiomyopathies. The VHS measured on the RL An approximately 5-y-old, 5.67 kg, intact female, New Zealand White views, but not VD and DV views, was significantly higher in the rabbit presented for red urine. She arrived as a 2-y-old retired cardiomyopathy group compared to the control group (P = 0.025). breeder from a vendor and was pair housed. The rabbit was assigned When divided into HCM and nonHCM group, only the nonHCM to an immunology study that involved regular whole blood group had significantly higher VHS values when compared to collection from the central ear artery. Examination of the pan controls (P = 0.035). When comparing the nonHCM group with the revealed dark red urine along with formed, hard, dry feces. A urine control group in the RL views and using the receiver operating dipstick test confirmed the presence of blood. Physical examination characteristic curve (ROC), the analysis suggested a VHS of 10.45 as a did not reveal any additional abnormalities. Based on the lack of cut-off value and yields an AUC of 0.78 with sensitivity of 83.3% and clinical signs other than hematuria, continued monitoring was specificity of 79.3%. When comparing all cardiomyopathies with the elected. A small amount of blood was noted in the urine the control group, analysis suggested a VHS of 10.25 as a cut-off value following day, but then subsided. Regular monitoring, including and yields an AUC of 0.64 with sensitivity of 60.5% and specificity of visual assessment and weighing, was continued for 2 mo and then 71.3%. In summary, VHS measured on the RL view may have some discontinued. Approximately 7 mo after resolution of the initial benefit in the clinical assessment identifying nonHCM clinical signs, the rabbit re-presented for marked hematuria with cardiomyopathies in rhesus macaques, but VHS did not replace the several blood clots present in the pan. On physical examination, the need for echocardiography as a cardiac screening tool in rhesus rabbit was lethargic with generalized pallor. Additional diagnostic macaques. tests were considered, but due to the rabbit’s age, euthanasia was elected. Postmortem examination revealed multiple, dark red-brown, P92 Unique Presentations of Burkholderia gladioli Infections in variably well-demarcated, small multinodular masses associated Several Strains of Immunocompromised Mice with the uterine horns. On histopathology, the endometrial masses were composed of numerous erythrocyte-filled vascular channels AJ Osborne*, SE Clark, H Atkins lined by well-differentiated endothelial cells separated in honeycomb-like fashion by thick seams of stroma, consistent with Comparative Medicine, Penn State Hershey, Harrisburg, PA vascular hamartomas. On the superficial aspect of some of these masses were large, dilated blood vessels, characteristic of Four strains of mice (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ [NSG], NOD. endometrial venous aneurysms. Vascular hamartomas are Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ [NRG], B6.129S(Cg)-Stat1tm1Dlv/J [Stat1- nonneoplastic, disorganized proliferations of vascular tissue that are /-]), and B6.129S7-Ifngr1tm1Agt/J [IFNγR-/-]) housed in a barrier facility thought to be congenital and typically do not cause clinical disease. developed unique and seemingly unrelated clinical signs; all affected The clinical signs observed in this case likely stem from an overlap of mice were experimentally naïve. Young NSG/NRG mice (n=49, mean the conditions of endometrial venous aneurysm and vascular age = 4 mo, +/- SEM 0.36) exhibited nonspecific clinical signs of hamartoma. Vascular hamartomas with complications from other moderate-severe lethargy, hunched posture, decreased body concurrent conditions should be considered as a differential condition, and pallor. In contrast to the NSG/NRG mice, the Stat1- diagnosis for rabbits presenting with hematuria. /- and IFNγR-/- mice (n=5) developed large subcutaneous abscesses on the head and neck. Liver, feces, and environmental samples were P94 Nasal Nematode Found in Meadow Voles collected for culture and PCR analysis. NSG/NRG mice had moderate-markedly enlarged livers (20/49, 40%) and spleens (17/49, B Stock*, JD Reuter, TT Mufford 35%). The livers contained multiple, variably sized, tan nodules throughout all lobes. Histology revealed necrotizing hepatitis (11/15, Office of Animal Resources, University of Colorado Boulder, Boulder, 73%), splenic and hepatic extramedullary hematopoiesis (15/15, CO 100%), glomerular histiocytosis (6/15, 40%), and metritis (5/10, 50%) with perivascular inflammation, suggesting hematogenous spread. In late 2018 there were numerous mysterious deaths in a meadow Differentials for these lesions included Mouse Hepatitis Virus, vole (Microtus pennsylvanicus) colony at a university. To determine Ectromelia virus, Salmonella spp., Clostridium piliforme, or other the cause of these deaths, a diagnostic necropsy was performed and infectious agents. Burkholderia gladioli was cultured from liver lesions tissues submitted to histology. The cause of death was linked to and subcutaneous abscesses and confirmed with 16S ribosomal RNA intestinal dysbiosis. However, we unexpectedly found an unusual sequencing. The IVC racks were sanitized and new breeders were nematode within the nasal passages that appeared to be reproducing. purchased, resulting in a dramatic reduction in B. Photomicrographs of the nasal cavity showed a cystic structure in the gladioli infections. B. gladioli is a ubiquitous, gram negative, aerobic mucosa that contained larvae, as well as eggs, indicating active bacterial rod associated with plant rot and respiratory infections in replication. The ova did not incite an inflammatory response, which immunocompromised humans, particularly those with cystic could negatively affect the health of the animal. The ova were about fibrosis. The current literature contains sparse reports of B. 70 um x 43 um bipolar ovoid shapes embedded in the nasal gladioli infections in immunocompromised mice, with a typical epithilium. Based on egg morphology, host, and location of cysts, the presentation of torticollis and rolling. Systematic testing of nematode was thought to be of the order Trichocephalida, family environmental samples is underway, but we suspect that drinking Capillariidae. More specifically, the parasite is likely of the water was contaminated during a period of time when the water genus Eucoleus, which is characterized by doubly operculated ova autoclave cycle failed routine quality testing. B. gladioli infection is a with pitted surfaces. The life cycle is usually direct, so the meadow potential differential for subcutaneous abscesses, hepatitis, and vole is likely the definitive host and not an intermediary. Because the

631 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 voles in this affected colony were wild-caught, they are at risk for a Due to some evidence that, similar to Dirofilariasis, Wolbachia sp play more diverse array of parasites. While we routinely screen rodents a role in the health and survival of the Brugia, the dog was first for pinworms, our routine detection methods (PCR, tape tests, fecal treated with a 30-dcourse of doxycycline (10mg/kg BID). This was floatation) failed to detect this nematode. Additionally, the voles were followed with a 5-d course of albendazole (50mg/kg q24hr), and not prophylactically treated during quarantine. Reports suggests that once weekly injections of ivermectin (400 mcg/kg) which continued fecal flotation is the best method for detecting this parasite as eggs for 1 y. Throughout treatment, and in the 8 mo following the are accidently swallowed, although this method failed to detect the completion of the ivermectin, blood was drawn regularly for parasite. Fenbendazole may be used to treat this infection, but the microfilarial counts. After 5 mo of ivermectin treatment, the hind effectiveness and exact treatment regimen remain unknown. The limb edema had visibly resolved and the microfilarial count was direct life cycle of this parasite enables it to persist in highly dense negative. The microfilarial counts remained negative for the 8 mo populations where animals are interacting with cage mates. following treatment. A necropsy was performed at the time of euthanasia and no gross pathology or signs of adult worms were P95 Optimization of a Novel Fetal Surgical Protocol in a Sheep noted. (Ovis aries) Model P97 Intracranial Biphasic Brain Tumor in a Naive Female A Cummins*1, M Connell1, S Duru2, M Oria2, JL Peiro2, C Doerning1 Cynomolgus Macaque (Macaca fascicularis)

1Veterinary Services, Cincinnati Children’s Hospital Medical Center, C Mano*1, MC Kundu1, H Jonassen2, J Mysore2, H Burr1 Cincinnati, OH; 2Center for Fetal and Placental Research, Division of Pediatric General and Thoracic Surgery, Cincinnati Children’s 1Veterinary Sciences, Bristol-Myers Squibb, New Brunswick, Hospital Medical Center, Cincinnati, OH NJ; 2Pathology, Bristol-Myers Squibb, New Brunswick, NJ

Sheep are a common model for reproductive and fetal research Three mo postarrival into an indoor facility, a naive, 2-y-old female owing to their size, docile temperament, and relative low cynomolgus macaque (Macaca fascicularis) of Mauritian origin was maintenance within a vivarium setting. Despite their established use reported for acute, seizure-like activity. The animal had been as a fetal research model, the reproducibility of fetal surgical evaluated 6 wk prior with no abnormalities noted on complete protocols can prove challenging. Multiple fetal and maternal factors physical, neurologic, and ophthalmologic examinations, complicate a straightforward protocol, such as unique nutritional and electrocardiography, pulse oximetry, and clinical pathology metabolic demands, appropriate selection of anesthesia and evaluation (CBC, serum chemistry, coagulation panel, and analgesia, refinement of surgical techniques when undergoing urinalysis). During examination, the animal was quiet and multiple survival surgical procedures, and unexpected complications. moderately responsive with decreased activity and sluggish Here we describe the various hurdles encountered during the ambulation. The animal was moderately dehydrated and exhibited implementation of a novel fetal myelomeningocele protocol at our intermittent whole-body shaking, absent convulsions, or rhythmic institution, and the solutions developed as a collaborative effort muscle twitching. No other abnormalities were noted and between veterinary and investigatory personnel. Ongoing subcutaneous lactated Ringer’s solution was administered. The refinements include perioperative supportive care of the ewe, animal was monitored throughout the day, with subsequent clinical surgical and anesthetic techniques to minimize fetal stress, preterm signs that included minimal responsiveness, slow circling, eyes delivery and resuscitation of neonatal lambs, and anesthetic support partially closed, and mouth open with tongue deviated to the right. of neonatal lambs undergoing magnetic resonance imaging (MRI). Brief seizure activity was directly observed causing the animal to be We emphasize the challenge of reproducibility in large animal unresponsive and recumbent. Diazepam was administered yielding surgical literature, the importance of interdepartmental collaboration, partial recovery, minimal responsiveness, and nystagmus. Due to and the dynamic nature of protocol refinement to optimize animal poor prognosis, the animal was euthanized. Clinical pathology welfare and produce successful, consistent research outcomes. parameters (CBC, serum chemistry) were within normal limits. Necropsy revealed a tan, irregular shaped, firm mass at the right side P96 Doxycycline, Albendazole, and Ivermectin Treatment of a Dog of the skull base near the temporal bone. The mass was with Peripheral Edema Caused by Brugia pahangi Infection approximately 2.5 cm X 3.5 cm, located within the dura mater, laterally compressing the brainstem and cerebellum, as well as the C Chambers*, E O’Connor cerebrum dorsally. Histologically, abutting and infiltrating a cranial nerve ganglion, the neoplasm exhibited biphasic cytology University of Missouri, Columbia, MO (polygonal/fusiform or small round cells). The differential diagnosis for this biphasic neoplasm includes medulloblastoma (desmoplastic/ Brugia pahangi is a lymphatic filarial nematode found in Malaysia, nodular-type), neuroblastoma, primitive neuroectodermal tumor Thailand, and Indonesia. In a research setting, it is used as a model to (PNET), mixed glioma, and meningioma, pending special and study the immune response to the related filarial worms, Wuchereria immunohistochemical stains. The occurrence of spontaneous brain bancrofti and Brugia malayi, which cause human cases of lymphatic neoplasms in cynomolgus macaques is rare, particularly in juveniles. filariasis and elephantiasis. Here, a small number of dogs are infected with B. pahangi, and serve as a host for the adult worm to produce P98 All Frogs Go to Heaven: Reused Tricaine Effectively microfilaria, which can be collected via blood draw for use in Euthanizes Xenopus laevis experiments. These dogs are usually asymptomatic. However, 1 dog developed clinical signs suggestive of peripheral lymphatic blockage, M Hoang, A Craig, S Ahmadi, DK Chu* and was subsequently treated for lymphatic filariasis. The subject was a 2-y-old, intact male beagle that was initially infected with L3 Comparative Medicine, Stanford University, Stanford, CA larvae of B. pahangi subcutaneously in the inguinal region. Approximately 3 mo later he presented with nonpainful, pitting African clawed frog (Xenopus laevis) is a household name in the field edema of the distal right hind limb, extending to just above the level of biomedical research and is a widely used model species in the field of the hock. Nuclear scintigraphy of the lymphatics showed evidence of developmental biology. While the American Veterinary Medical of a lymphatic blockage at the level of the popliteal lymph node. Association states tricaine methanesulfonate solution immersion as Interestingly, the microfilarial count was also very low, and the dog an accepted euthanasia method for many aquatic species, making was therefore removed from the original study. Lymphatic filariasis fresh batches of tricaine for each cohort can be labor intensive. In an is notably very difficult to treat and eliminate in both humans and effort to create more efficiency in our scaled-up frog euthanasia animals, therefore a followup project was developed to evaluate the procedures, we evaluated tricaine submersion with both fresh and efficacy of a multimodal treatment approach to clear the infection. reused buffered tricaine solution (5 g/L). In our study, 10 adult

632 Abstracts of scientific papers 2019 AALAS National Meeting

female frogs were randomly assigned to each group. Groups 1, 2, and webbing, and mild buoyancy compromise. Due to the progression in 3 were immersed in freshly made buffered tricaine, once used severity of the clinical condition over the next 6 d, this frog was tricaine, and twice used tricaine, respectively. Following 1-h euthanized. Differential diagnoses included primary infection, immersion, frogs were removed from tricaine and monitored hourly trauma with associated infection, and altered water quality for spontaneous activity and pedal withdraw reflexes. At end of the parameters. Diagnostic tests included cultures, blood work, fifth hour a celiotomy was performed to assess ventricular necropsies, and histopathology. Cultures and bloodwork were contractions, if any. Solution temperature and pH were monitored. unremarkable. On gross examination, many gas-filled bubbles were After 1-h immersion in tricaine solution and throughout 5 h seen in the interdigital webbing of the hind limbs. The remainder of postremoval from tricaine, no frogs in any group displayed signs of the gross examination was unremarkable. Histopathology revealed spontaneous activity or withdraw reflexes. Mean ventricular small to large empty spaces (interpreted as gas-filled based on gross contraction rate at end of fifth hours for Group 1, 2, and 3 was 11.2, appearance) expanding the deep dermis. In the interdigital webbing, 7.4, and 9.5 contractions per minute, respectively; all significantly the epidermis contained multiple single cells interpreted as necrotic lower values than the referenced normal resting Xenopus heartrate of keratinocytes. Based on the gross lesions, histologic skin changes, 60 per minute. Solution temperature and pH did not fluctuate. From and clinical history, the primary cause of morbidity was determined these studies, we concluded that tricaine submersion can be to be gas supersaturation. The clinical signs were likely secondary to effectively used for at least 3 times, along with secondary acute water temperature increases, and chronic air leaks in water confirmation for Xenopus euthanasia. These findings will help us transport piping. Gas bubble disease occurs when water is refine our euthanasia protocols for our frog colonies. supersaturated with gases. Saturation can be altered by increasing tank temperatures, increased water pressure via leaks in system P99 Propofol Overdose Effectively Euthanizes Zebrafish Danio( piping, or altered salinity. Xenopus laevis are susceptible to gas bubble rerio) disease. Preventative measures such as introducing degassers, maintaining communication with municipal water suppliers/facility DK Chu*1, C Pacharinsak1, K Jampachaisri2 operation managers, and monitoring saturation/dissolved oxygen should be implemented in an aquatic facility. 1Comparative Medicine, Stanford University, Stanford, CA; 2Faculty of Science, Naresuan University, Mueang Phitsanulok, , Thailand P101 Evaluation of Therapeutic Approaches for the Treatment of Spironucleus muris in Mice Tricaine and hypothermia using an ice bath are common zebrafish euthanasia methods. While tricaine poses human health hazards GB Voros*, DD Andrews, GL Dobek during its preparation, hypothermia is only recommended for some fish, specifically small tropical or subtropical fish. The aim of this Comparative Medicine, Tulane University, New Orleans, LA study is to explore an alternative euthanasia method. Here, we evaluated propofol overdose for zebrafish euthanasia. We Spironucleus muris is a protozoan pathogen of mice that rarely causes hypothesized that a propofol overdose at 40 mg/L with a 10-min clinical signs of disease, but can result in morbidity and affect submersion euthanizes zebrafish as effectively as a 20- or 30-min research outcomes, especially in the case of gastrointestinal and submersion. Thirty adult wild-type AB zebrafish (Danio rerio) were immunological research. The current recommendation to rid infected randomly divided into 3 groups of 10 fish each. All fish within each colonies of this pathogen is to test and cull affected animals. In some group were simultaneously immersed into 40 mg/L propofol for 10, cases this is not feasible due to irreplaceable genetic lines, valuable 20, or 30 min then monitored for recovery. Time to loss of muscle aged animals, or other unique models. The purpose of this study was movement, operculation cessation, and startle reflex (to tank tap) to evaluate fenbendazole and metronidazole for the treatment of were recorded. Signs of distress (piping, erratic swimming, twitching, mice naturally infected with S. muris. We chose these drugs due to regurgitation), if any, were recorded. At the end of submersion their effectiveness in treating Giardia, a protozoal parasite of the same period, fish were moved to a recovery tank and the number of fish Family as S. muris. Five CD1 mice naturally infected with S. recovered up to 180 minutes was recorded. No fish showed signs of muris were used per group. Groups were treated with fenbendazole stress. Time to loss of muscle movement (19-22 seconds) and 50 mg/kg via sucralose-flavored medication delivery gel, operculation cessation (55-66 sec) were not significantly different metronidazole 500 mg/kg via sucralose-flavored medication delivery between groups. Loss of startle reflex was not significantly different gel, fenbendazole 50 mg/kg and metronidazole 500 mg/kg via between the 20- and 30-min groups while fish in 10-min group did sucralose-flavored medication delivery gel, fenbendazole 50 mg/kg not lose reflex through the 10-min period. For recovery, none of the via oral gavage, untreated sucralose-flavored medication delivery gel fish in 20- or 30-min group recovered while 70% of fish in 10-min as a control, and water via oral gavage as a control daily for 4 wk. submersion group recovered. This study demonstrates that propofol Retesting of individuals was done via submission of fecal samples for overdose at 40 mg/L at 10-min immersion does not euthanize PCR on a weekly basis. At the end of the 4-wk period, mice were zebrafish as effectively as a 20- or 30-min immersion. We conclude euthanized and gastrointestinal samples were sent for histological that propofol immersion at 40 mg/L for at least 20 minutes examination. Our results showed that no treatment of fenbendazole, effectively euthanizes zebrafish. metronidazole, or a combination of therapeutics were effective in the treatment of S. muris. Further studies are needed to see if higher P100 An Un-frog-gettable Case Report: Gas Bubble Disease doses are effective and to optimize route of administration. in Xenopus laevis P102 Tip Matters: Jugular Vein Catheter Patency in Sprague- ER Feldman*1, WO Williams1, M Forzán2 Dawley Rats

1Center for Animal Resources and Education, Cornell University, H Roeder* Ithaca, NY; 2Department of Population Medicine and Diagnostic Sciences, Cornell University, Ithaca, NY PKPD, BioAnalytical Systems Inc., West Lafayette, IN

Three adult, female, Xenopus laevis were reported for “red legs” 3 d Surgical implantation of jugular vein catheters (JVCs) facilitates after a system malfunction altered the housing tank temperatures. central venous access in preclinical models. Use of JVCs can enhance The frogs were IACUC-approved for surgical oocyte collection, and animal welfare and potentially reduce the number of animals individually housed in flow-through tank systems. Two frogs needed, along with refining study design to get better, translatable presented with acute hyperemia, skin sloughing, bloating, and data. Patency, or bidirectional flow, defines the functional life of a lethargy, and were euthanized due to poor prognosis. The third frog JVC; therefore, it is important to examine potentially contributing presented with minor hyperemia of the right hind interdigital factors, such as catheter tip shape and maintenance. Rat JVCs are

633 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 widely accepted and used. However, design and manufacturing Common marmosets (Callithrix jacchus) are a popular nonhuman process vary among institutions with limited scientific evidence primate model for assisted reproductive technologies and genetic surrounding the efficacy of either factor. This study compares engineering due to their small size, ease of handling, and robust patency of JVCs from 3 different manufacturers. Each JVC is made of reproductive capacity. The creation of transgenic infants requires a 3-french polyurethane; 2 have a rounded tip, and 1 has a bullet tip. multistep process with an end goal of live birth of offspring after Fifteen male 300g Sprague Dawley rats were randomly assigned to 1 implantation of genetically modified embryos into recipient females. of 3 groups (n=5) and catheterized via the right jugular vein. Catheter We noted an increased rate of dystocia requiring cesarean section in patency was evaluated every 3-4 d for 28 d. Catheters were deemed our marmoset colony after assisted reproductive procedures when fully patent if blood was withdrawn during the initial attempt. If compared to natural mating. To evaluate the potential risk factors for blood was not immediately withdrawn, 0.1 ml saline infusion was the increased incidence of cesarean sections, we reviewed the medical followed by a second attempt. If successful, catheters were records of all marmosets giving birth within a 2-y period. During the considered partially patent. If unsuccessful, catheters were study period, the rate of cesarean section was significantly higher considered nonpatent. After 28 d, 100% of one rounded tip, 60% of following embryo transfer (30.7%, 4/13) as compared to natural the other rounded tip, and 80% of the bullet tip catheters were conception (2.6%, 2/76) [P = 0.0037]. The overall rate of cesarean considered patent. Bullet-tip catheters showed repeated instances of section was 6.7% for all births during the study period (6/89). partial patency and had the first instance of nonpatency. In Indications for cesarean section included straining, abnormal posture conclusion, this pilot study indicates rounded-tip design is superior and gait, fetal entrapment, and cage mate aggression. Gestational to bullet-tip design in maintaining catheter patency. length following embryo transfer was longer (145 d) for animals requiring a cesarean section as compared to natural birth (143 d). P103 Screening and Identification of Mouse Kidney Parvovirus in Gestational length was calculated using embryo stage at the time of Association with Chronic Renal Nephropathy in a Colony of implantation. Cesarean section resulted in at least 1 live-born infant Immunocompromised Wsh/Wsh Mice in 71.4% of cases (5/7), of which the majority of infants (71.4%, 5/7) were fostered to a lactating surrogate female. The results of this S Carrasco, Z Ge, Y Feng, S Muthupalani, V Bakthavatchalu, D study highlight the need for careful observation of marmosets Annamalai, JG Fox* following embryo transfer as cesarean section may be required. Further research will determine additional risk factors for the need to Division of Comparative Medicine, Massachusetts Institute of perform cesarean sections in marmosets as well as post-operative Technology, Cambridge, MA fertility and infant survival following cesarean section.

The occurrence of spontaneous nephropathy with intranuclear P105 Pharmacokinetics and Efficacy of Oral Mirtazapine in Guinea inclusion bodies in the renal tubular epithelium of laboratory mice Pigs (Cavia porcellus) was historically described in immunompromised mice without ascribing definitive etiology. However, recent evidence demonstrated JD Ayers*1, E Stietzle4, M Ellis2, J Kim3, LV Kendall1 a novel mouse kidney parvovirus (MKPV) was the cause of this inclusion body-associated nephropathy. We had recently reported on 1Laboratory Animal Resources, Colorado State University, Fort the occurrence of a sudden increase in morbidity and mortality in a Collins, CO; 2Veterinary Medicine and Biomedical Sciences, Colorado colony of immunocompromised Rag2-Il2rγ knockout Wsh/Wsh mice, State University, Fort Collins, CO; 3Veterinary Medicine, Ohio State which was attributed to pks+ E. coli-induced sepsis. Interestingly, a University, Columbus, OH; 4Veterinary Medicine, Kansas State subset of these mice (5/15, 7-22-mo-old) also had markedly small, University, Manhattan, KS pale-tan indented kidneys. On histopathological examination, the renal cortical and medullary parenchyma was characterized by Guinea pigs are cecal fermenters requiring frequent and consistent multifocal to coalescing areas of tubular degeneration/necrosis, feed intake to ensure normal gut motility. Transport, age-related tubular loss and atrophy, ectatic tubules with amphophilic casts, mild disease, diet changes, and other sources of chronic stress can reduce to moderate interstitial fibrosis and low numbers of interstitial their appetite, leading to gastrointestinal stasis which can be life inflammatory aggregates chiefly macrophages and neutrophils. threatening in this species. Mirtazapine, a tetracyclic antidepressant, Many affected tubules were lined by swollen, vacuolated tubular is used in dogs and cats to treat nausea and inappetance, and has epithelial cells with pale-eosinophilic, granular cytoplasm, and been shown to increase feed intake in cats. It has anecdotally been karyomegalic nuclei with marginated chromatin and large used as an appetite stimulant in guinea pigs, but a therapeutic dose amphophilic intranuclear inclusion (viral) bodies. These changes of mirtazapine has not been established for this species. An initial were consistent with a moderate to severe chronic tubulointerstitial study using 6 healthy male guinea pigs administered mirtazapine at nephropathy associated with the presence of atypical, intranuclear 1.88, 3.75, or 7.5 mg orally once daily for 4 d, in a crossover design, inclusion bodies within the renal tubular epithelium in a manner showed no significant differences in weight gains, feed intake, or similar to those observed in association with MKPV. Retrospective fecal output as compared to 3 male guinea pigs of similar age given evaluation of additional mice with the same genotype indicated an saline only. Pharmacokinetic analysis showed peak plasma levels additional 12 mice (3.5-10-mo-old) had histopathological lesions being reached at 30 min and returning to 0 at 8 h, demonstrating that consistent with MKPV-induced nephropathy. To further confirm our dosing every 24 h is not appropriate for this species, which etiological diagnosis, PCR was performed on formalin-fixed and supported the lack of clinical response. A follow up study was paraffin-embedded kidney sections using primer sets targeting NS1 performed in another set of 6 healthy male guinea pigs with every 8 of MKPV. All the mouse kidney sections tested positive for MKPV. h dosing at 1.88mg orally for 4 d, compared to 3 saline only controls. Sequence analysis of the PCR products shared 99% identity with Feed weights demonstrated a significant increase in feed intake for MKPV DNA detected in the MKPV strain MSKCC. The results the mirtazapine group versus the saline group, but no significant demonstrate the existence of MKPV in our facility and provide difference in weight gains. This study shows that mirtazapine must additional molecular and histopathological evidence for the presence be dosed appropriately for the species, in this case every 8 h, in order of this novel virus in mice originating in the U.S. to be used as an appetite stimulant in male guinea pigs. Further studies are warranted to examine the effects in females, and to P104 Embryo Transfer Increases Incidence of Cesarean Section in determine if higher doses would cause a greater increase in feed Common Marmosets intake, particularly in clinically ill animals where improving appetite is imperative. JL Haupt*, MA Burns

Massachusetts Institute of Technology, Cambridge, MA

634 Abstracts of scientific papers 2019 AALAS National Meeting

P106 Multifocal Discrete Masses in a Genetically Modified Mouse mandatory rederivation for all lines in barrier housing. Testing was accomplished by adding MKPV PCR to all samples submitted for J Lee*, BF Taylor, E Nunamaker routine quarterly health testing to a major animal health laboratory, which occurs at all vendor-defined health standards. In IVC housing Animal Care Services, University of Florida, Gainesville, FL locations, 1 animal per rack side was tested. In wire-top static cage locations, at least 5 animals per room were tested, and at least 1 A cage of 2-mo-old, male NOD.Cg-Ncf1m1J/MxJ breeder mice animal was tested from each gnotobiotic isolator. Animals tested was reported for fighting and mild tail trauma. On examination, were dirty bedding sentinels as well as line animals, with a minimum multifocal pinpoint-crusted fight trauma on the tail was observed age of 10 wk, and a minimum dirty bedding exposure of 6 wk. All on all 3 mice in the cage. In addition, the mice had multifocal commercial colonies, including those at Asian production partners, small, firm, raised, superficial masses on their tails. One of the mice tested MKPV-negative. also had a discrete, raised, firm, superficial mass on the cranial aspect of the right front manus. None of the masses expelled P108 Defining and Validating the Specific-pathogen Free State discharge and no treatment was prescribed. The mice were of Xenopus Using qPCR Assays monitored for clinical progression of the masses. Three days after initial presentation, additional superficial skin masses were seen K Bowes*, C Hensley, S Feldman on the dorsal metatarsus, ears, ventral neck, flanks, and lateral muzzle. Differential diagnoses for the multifocal masses included Center for Comparative Medicine, University of Virginia, pyogranulomatous inflammation secondary to bacterial infection, Charlottesville, VA sterile pyogranulomatous inflammation, or neoplasia. Based on the clinical presentation and the progression of the masses, euthanasia Colonies of valuable inbred and transgenic laboratory was elected. Gross necropsy was performed and samples were reared Xenopus frogs constitute naïve populations of animals obtained for cytology, histopathology, and bacterial culture. On gross susceptible to some opportunistic infectious diseases. Therefore, necropsy, enlarged spleens and peripheral lymphadenopathy of the it is prudent to characterize any new animal acquisitions to an mandibular, inguinal, and popliteal lymph nodes were observed. existing colony as a biosecurity measure to preclude the concurrent A thick, yellow, purulent discharge was aspirated from the masses. introduction of an infectious disease microorganism with the Cytologic examination showed a homogenous population of new animals. Since it is not cost effective to test for all diseases neutrophils. Histopathology revealed neutrophilic granulomatous of Xenopus frogs, we have defined a subset of prevalent infectious inflammation. Bacterial culture of the discharge was positive disease microorganisms and developed 10 qPCR assays to detect for Staphylococcus xylosus. Based on the test results, a diagnosis these disease-causing agents. The specific pathogens in our test of pyogranulomatous inflammation secondary to Staphylococcus panel were selected from relatively recent publications as causing xylosus infection was made. The lesions observed were likely morbidity and/or mortality in Xenopus laevis and/or X. tropicalis. In associated with the genetic alteration of the Ncf1 gene in this addition, many of the pathogens in our panel are zoonotic diseases mouse model. Abnormalities of the Ncf1 gene leads to dysfunction placing frog aquarists at risk for acquiring infections. These 10 of the NADPH oxidase protein in neutrophils, thereby causing assays do not represent a comprehensive list of infectious diseases impaired pathogen destruction by neutrophils and pyogranuloma of Xenopus frogs, and so frogs devoid of the infectious diseases in our formation. Because NOD.Cg-Ncf1mJ1/MxJ mice are susceptible test panel are characterized as “specific-pathogen free.” Three of the to pyogranulomatous inflammation, aseptic practices and special assays broadly detect many species within their respective genera husbandry care should be implemented to prevent inadvertent (i.e. Ranavirus, Chlamydophila spp and Cryptosporidium spp). infections. P109 Removing the Blindfold in Blind Intubation: A Summary of P107 Prevalence of Mouse Kidney Parvovirus at a Major Rodent Tips and Tricks for Successful Rabbit Intubation Vendor K Szymczyk*, S Gabriel, T Worlds, CZ Cannon J Ramirez-Komo*1, MJ Crim2, M Keith3, PL Roesch4 Becton Dickinson, Research Triangle Park, NC 1Veterinary Sciences, Taconic Biosciences, Germantown, NY; 2IDEXX BioAnalytics, Columbia, MO; 3Diagnostic Services, Taconic Achieving a patent airway during anesthesia is critical to prevent Biosciences, Rensselaer, NY; 4Diagnostics and Environmental airway obstruction and to deliver gas anesthetics. Intubation in Biosecurity, Taconic Biosciences, Rensselaer, NY rabbits can be difficult due to their unique anatomical features. It is the hope of the authors to share techniques that can be help A novel parvovirus has been recently described which causes clinical increase successful intubation in the rabbit. A variety of techniques disease in immunocompromised mice. Mouse kidney parvovirus and approaches are discussed including position of the animal, (MKPV, also known as mouse chapparvovirus, MuCPV) is highly position of the intubation tube, methods to monitor progress and genetically divergent from other mouse parvoviruses, such as minute clinical and practical assessments of successful placement. Common virus of mice (MVM) and mouse parvovirus (MPV). Viral infection complications and potential solutions will also be examined. The results in inclusion body nephropathy and renal fibrosis, leading to techniques discussed were used to train research staff the art of chronic renal disease and eventually renal failure in older severely rabbit intubation with adequate surgical plane of anesthesia and safe immunodeficient mice. MKPV is widely distributed among research recovery. This has resulted in individual staff members developing institutions in the United States, Canada, Europe, the Middle their own, unique method to intubate rabbits using these tools to East, and Australia based on a real-time PCR assay that has been aid in successful placement. Success criteria include the ability to validated to detect genetically divergent strains of MKPV. MKPV repeatedly intubate rabbits, confirm tube placement in the trachea, can be detected in a wide array of sample types, and approximately and maintain a surgical plane of anesthesia with uneventful 4.2% of mouse fecal samples, 5.1% of environmental samples, and anesthesia recovery. Blind intubation in the rabbit is challenging but 5.4% of transplantable tumors have tested positive to date. Mouse can be achieved using specific approaches and techniques learned feces are often soaked with urine, making feces an effective and in this program The ability to understand the anatomy of the rabbit easily collected cage-level sample. As a major vendor, we sought airway system combined with the use of practice animals for training to determine the prevalence of MKPV in our mouse colonies. successfully led to safe and effective rabbit anesthesia outcomes. Our commercial rodent colonies comprise 78 health reporting groups which include open wire-top static cage or individually ventilated cage (IVC) barrier facilities and gnotobiotic isolators. We hypothesized our colonies to be free of MKPV due to the practice of

635 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P110 Surgical Intervention of Hepatic Hemorrhage in a morbidity or moribundity, helped to determine preemptive Cynomolgus Macaque (Macaca fascicularis) euthanasia prior to animals becoming moribund, provide better communication between husbandry technicians and veterinary staff K Yenney*, C Cruzen, B Megrath, B Knapp, D Benedict and ultimately improve animal welfare by minimizing pain and distress associated with GvHD, the effects of TBI and the Scientific Services, AltaSciences Preclinical, Ellensburg, WA humanization process.

Immediately following a liver biopsy of a young, adult, male P112 Use of a Handheld Ketone Meter in the Management of cynomolgus macaque (Macaca fascicularis), a significant hepatic Diabetes Mellitus in Rhesus Macaques (Macaca mulatta) hemorrhage was observed via ultrasound. Abnormalities were detected in both heart rate and blood pressure. An intravenous KE Killoran*, AE Sarfaty, JL Asher, SR Wilson, PC Smith catheter was placed in the right saphenous vein and approximately 120mL of 0.9% NaCl was administered. A 6-cm midline abdominal Comparative Medicine, Yale University School of Medicine, New incision was made, and the cranial abdomen was packed with gauze. Haven, CT Approximately 40mL of frank, unclotted blood, including a large blood clot, was removed and site of hemorrhaging was identified. A Diabetes mellitus is a common condition in aging rhesus macaques 2cm square of sterile compressed sponge was placed and the (Macaca mulatta). In addition to blood glucose and glycated abdomen was closed with 2 layers of suturing: simple continuous in hemoglobin measurements, blood and urine ketone measurements body wall and subcuticular in skin. Postoperative hematocrit showed are essential for optimal management of this chronic disease. As in significant anemia (Hct: 18% PCV, Hb 6.1 g/dL). The animal other veterinary species, the presence of ketonemia or ketonuria recovered from sedation slowly but was bright alert and responsive often indicates that the individual has an uncontrolled comorbidity upon placement in the home cage. Upon observation the following or requires further diagnostic or therapeutic care. Ketotic animals day, herniation was noted at the incision site and when placed into a must be identified and treated before the metabolic derangement procedure cage (restraint device), immediate dehiscence of the progresses to diabetic ketoacidosis (DKA) as intensive care of surgical incision occurred, resulting in a body wall hernia and macaques is difficult and frequently unsuccessful. Ketone test strips subsequent gastric protrusion. Surgical repair was successful. No do not allow for the detection of betahydroxybutyric acid (BHBA), additional hepatic hemorrhage was noted. Anemia was still noted at the first ketone produced when an animal begins to undergo this time (Hct 15% PCV and Hb unreadable). The animal was ketogenesis. However, newer handheld ketone meters specifically observed 3 times a day for 1 wk and activity level increased each day. measure BHBA and can help to identify animals in the earliest stages The surgical site healed successfully, the animal’s appetite was good, of ketogenesis. We use a handheld ketone meter in our colony of and urine/fecal output were within normal limits. No other aged diabetic rhesus macaques, but no published reference ranges complications were noted, and the animal was taken to scheduled exist for blood ketones in the rhesus macaque. Therefore, we necropsy 13 wk later. The pathologist noted that the median and left established a normal reference range for blood ketones using lobes of the liver adhered to abdominal cavity and to each other. approximately 90 macaques of varying ages, health statuses, and These adhesions were likely the result of surgery within the glycemic conditions. This reference range will be an invaluable abdomen. Given the success of this surgical intervention, it is a viable resource for the management of diabetic macaques and will enable option that should be considered for this type of adverse event. prompt detection and treatment of DKA in the rhesus macaque.

P111 Humane Endpoint Refinement for Total Body Irradiation and P113 Rats! Something Ain’t Right Humanization of NCG Mice L Anderson1, J Anderson2, ML Martin*1 KY Jen*, J Rowe, S Festin 1Veterinary Services Program, Walter Reed Army Institute of Charles River Laboratories, Wilmington, MA Research, Rockville, MD; 2Veterinary Services Program, Naval Medical Research Command, Silver Spring, MD Humanization of mice has proved to be a promising avenue for translational research of human hematopoiesis and immunity. The A 6-mo-old, male Sprague Dawley rat (Rattus norvegicus) was utility of immunodeficient strains which lack adaptive and/or innate examined for lethargy, hunched posture, and periocular porphyrin immune function such as the NOD-Prkdcem26Cd52Il2rgem26Cd22/NjuCrl staining. Four days previously, the rat began receiving daily (NCG) mice have also allowed for better engraftment rates of intraperitoneal injections of a vehicle (control animal) as part of an humanized cells. Developed using CRISPR/Cas9, the NCG mouse individually housed PTSD study. On physical examination, the rat carries a human-like variant of the Sirpa (SIPR a) gene which, in part, was noted to be lethargic, approximately 8% dehydrated, had permits xenotransplantation of immature hematopoietic stem cells. significant porphyrin staining around the eyes and nose, was The process of humanization however can carry with it side effects of hunched in posture, and tender on abdominal palpation. At that time irradiation dose and the eventual onset and development of 2 other animals were noted to have similar clinical signs. Differential xenogenic graft-versus-host-disease (GvHD). GvHD in mice typically diagnoses included acute stress, acute abdomen, or neoplasia. presents as unthriftiness, hunched posture and weight loss, fur loss, Treatments comprised of hydrogel and subcutaneous fluids, reduced mobility, or tachypnea. Often, total weight loss alone is used nevertheless there was no response and all 3 animals were as a parameter for end point criteria without consideration of other diagnostically necropsied. Common necropsy findings included parameters and despite widespread use of humanized mice, humane serosanguinous fluid in the abdominal cavity, hemorrhages on the endpoint criteria for humanization have not been clearly outlined in serosal surfaces of the intestines, and fibrin stranding along the the veterinary science and humanized mouse literature. To address intestines, liver, and abdominal cavity wall. Histopathology of the this issue, we developed a cageside scoring system specifically for liver showed multifocal areas of necrosis and inflammation; a foreign NCG mice following the process of humanization with human body (hair shaft) was found as the core of 1 liver lesion, with peripheral blood mononuclear cells (hPBMC) or human CD34+ cells surrounding inflammation and bacterial components. Culture of the which can include total body irradiation (TBI). This scoring system vehicle and all of its individual components showed no growth. included criteria surrounding overall appearance, physical exam However, culture of the serosanguinous abdominal fluid parameters, behavioral scoring, and body condition scoring. Using revealed Proteus mirabilis, which is described as a ubiquitous this method at our institution, we were able to calculate a cageside bacterium in the environment. Opportunistic infection with P. cumulative score, in addition to total body weight loss criteria to mirabilis resulting in disease has been observed in both provide a quantitative measure and refinement to humane immunocompetent and immunodeficient laboratory rodents. This endpoints. This, in comparison to a subjective assessment of case report highlights the importance of proficient intraperitoneal

636 Abstracts of scientific papers 2019 AALAS National Meeting

injection techniques to minimize the possibility of iatrogenic foreign additional dark retreat areas for the animals. Vivarium personnel body and environmental contamination of study animals. should be aware of environmental effects of light on animal health and welfare and the potential for inappropriately high levels of light P114 Evaluation of Medicated Gel as a Supplement to Providing intensity to impact interpretation of safety studies, especially in Acetaminophen in the Drinking Water of Sprague-Dawley Rats albino rodents. after Surgery P116 Liquid Diet-induced Bloat and Aspiration Deaths in Sprague- LE Riddle*, NL Rowley Dawley Rats Following Roux-en-Y Gastric Bypass

Center for Laboratory Animal Medicine, Uniformed Services LA Stewart*1, K Grimsrud1,2 University of the Health Sciences, Bethesda, MD 1Mouse Biology Program, University of California, Davis, Davis, Numerous studies have tested the suitability of specific analgesics to CA; 2Department of Pathology and Laboratory Medicine, University ameliorate postprocedural pain in laboratory rodents with varying of California, Davis, Sacramento, CA results. The drug selected and the route of administration are 2 major factors in whether an analgesic may provide sufficient, reliable pain Patients undergoing bariatric surgical procedures such as Roux-en-Y relief. Acetaminophen is an easily obtainable analgesic that can be gastric bypass (RYGB) are typically placed on a liquid diet pre- and self-administered to rodents in both water and gel products. This post-operatively to reduce strain on healing tissues and improve study compared the consumption of acetaminophen-treated water surgical outcomes. Human liquid diet products are regularly used in and/or gel offered to male Sprague Dawley rats following a sham laboratory settings when managing rodent bariatric models. Sprague- surgical procedure to determine if there was a preference for Dawley (SD) rats (n=40, male, 12-wk-old) were placed on 60% high acetaminophen delivery and to estimate whether the rats ingested a fat diet (HFD) for 4 wk to induce a state of obesity (DIO) prior to therapeutic dose of acetaminophen. Rats were assigned to 1 of 3 RYGB (n=35) or sham (SH) (n=5) surgical intervention. Rats were to treatment groups postsurgery based on the drug delivery system be introduced to liquid diet 24 h prior to surgery and maintained on provided: acetaminophen in water (AW), acetaminophen in gel (AG), it for 8 d postoperatively. The first 10 rats (7 RYGB, 3 SH) to undergo and acetaminophen in water and gel (AWG). Body weight surgery were given a human liquid diet product. Significant bloating measurements were taken 1 day prior to surgery (day 1) and for 3 was observed perioperatively in the upper gastrointestinal (GI) track, days postoperatively (days 1-3), while water and gel weights were resulting in an approximate 3-4x increase in overall stomach size. measured over 3 days. All 3 groups ingested acetaminophen doses During the first 48 h postoperative, 85% of the RYGB animals (n=6 of that were higher than the 200 mg/kg targeted therapeutic dose, with 7) exhibited acute respiratory issues including coughing, open the AW group ingesting the lowest average daily dose (350.9 mg/kg), mouthed breathing, and wet/raspy breathing sounds, as well as and the AG group ingesting the highest average daily dose (619.6 drooling, porphyrin staining, inappetence, and lethargy. Animals mg/kg). There was no significant difference among the groups for were euthanized and gross necropsy showed intact surgical sites, the average daily acetaminophen dose ingested. The findings of this significant bloating throughout the GI including foamy liquid in the study suggest that water and/or gel could be used by rats for self- esophagus, and mottled lungs consistent with aspiration pneumonia administration of oral acetaminophen as a postprocedural analgesic. in 83% (n=5 of 6). We suspected the human liquid diet aid may be causing the bloat and immediately switched over all remaining P115 Phototoxicity Identified in Rats in a Newly Renovated Animal animals to another brand of human liquid diet, which had previously Facility been used in a small pilot without incident. Perioperative bloat and mortality were considerably reduced after the diet change with only LD Meunier*1, DF Holliday3, J Caverly Rae2 6% (2 of 30) of the remaining rats euthanized for nonimpaction related aspiration pneumonia. Numerous ingredients and nutrient 1Global Laboratory Animal Medicine, GlaxoSmithKline, Collegeville, vary between the 2 liquid diets, therefore the etiology responsible for PA; 2Translational Medicine & Comparative Pathobiology, inducing bloat in HFD DIO rats remains unclear. This scenario GlaxoSmithKline, Collegeville, PA; 3In Vivo Sciences & Delivery, provides evidence that rats fail to metabolize the first human liquid GlaxoSmithKline, Collegeville, PA ddiet in a manner that supports healthy outcomes following bariatric procedures. We recommend using the second product or other liquid Outer retinal atrophy, characterized by a decrease in thickness of the diets proven safe in rats for procedures requiring animals to be on a outer nuclear layer of the retina, was observed during liquid-diet regimen. histopathological analysis of a 7-d rat safety study. This finding occurred at all dose levels, including control, indicating that the P117 Normal Blood Chemistry Reference Ranges for the Common lesion was not test-article related. As outer retinal atrophy can be Marmoset (Callithrix jacchus) Generated by a Point-of-Care Blood induced by exposure to intense light in rodents, particularly albino Analyzer animals, this finding immediately prompted investigation into potential sources of light exposure. The male Wistar Han rats LA Quinn*1, CH Toolan1,2, SC Artim1, JG Fox1, MA Burns1 originated at a vendor where they had each undergone surgery. Upon receipt at our facility, they were housed in a single holding 1Department of Comparative Medicine, Massachusetts Institute of room then transported to a procedure room for up to 8 h on study Technology, Cambridge, MA; 2University of Pennsylvania School of days. Our vivarium has recently been renovated and installed with Veterinary Medicine, Philadelphia, PA light-emitting-diode (LED) lighting. Light readings in the holding and procedure rooms were set at acceptable light levels according the The common marmoset (Callithrix jacchus) is a valuable animal model the Guide prior to animal occupancy. However, readings completed for multiple areas of biomedical research, including neuroscience and after the abnormal ocular findings were reported to be high (~120 immunology. Due to the growing interest in use of the common ft-candles) in the procedure room. Further investigation revealed that marmoset as a model, it is imperative that valid reference ranges be the dimmer switch installed in the room had been adjusted beyond established for all health monitoring parameters. Point-of-care blood acceptable levels of light intensity even though preventative analyzers are commonly used perioperatively, as well as during work measures were put in place. The vendor also reported high light up of both emergency and routine procedures. These devices are levels (1920 ft-candles) in the area where the rats had undergone capable of measuring blood parameters including liver and kidney surgery. Thus, it could not be determined if the inappropriate light function, acid base balance, blood gas levels, and serum electrolytes exposure occurred at the vendor or within our facility. Additional levels using only 2-3 drops of whole blood. In emergent clinical cases, procedures have been put in place to ensure maximum acceptable analysis of point-of-care blood parameters is critical for case workup light levels are not exceeded in our vivarium. We are also providing and health monitoring, but normal reference ranges for i-STAT

637 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 parameters for marmosets have yet to be established. Here we P119 Pain Assessment via Consumption of Yogurt Treats in Rats present a retrospective study of i-STAT values and normal reference Treated with Sustained-release Meloxicam ranges generated from healthy marmosets. The sample group was limited to healthy marmosets with blood drawn preoperatively for J Kim*, LV Kendall elective procedures. Reference ranges were generated and evaluated by sex (16 males, 33 females) and age, with 22 juvenile to adult (less Laboratory Animal Resources, Colorado State, Fort Collins, CO than 5 years) and 27 older adult to aged (5 years and older) marmosets included in the study. Statistical software was used to Minimizing pain in laboratory rats and its undesirable physiologic generate mean, standard deviation, relative standard deviation and behavioral impacts on biomedical research are necessary in (RSD), and 5%-95% percentile reference ranges for all parameters. science and medicine. However, the appearance and degree of Tests were performed to determine the treatment effect of age and pain is frequently ambiguous in rats. Recent advancements in pain sex. Statistically significant differences were noted between male and assessment (e.g. rat grimace scale, gait, and automated analyses) female median values for hematocrit, hemoglobin, and blood urea have limitations, including altered behaviors in response to nitrogen (BUN), however the magnitudes of the treatment effects humans and considerable financial investments. We evaluated food were small enough to be clinically irrelevant. The RSD of certain motivation in association with a cageside behavioral ethogram to parameters, including base excess of extracellular fluid (134.9%), enhance pain assessment in rats. Several food treats were evaluated glucose (34.2%), and BUN (29.8%) were high compared to other to determine which was highly valued, including sweetened parameters, indicating that clinicians may expect to see greater hazelnut cocoa spread, fruit jewels, and yogurt treats. Value was spread in these values. With this study, we have generated the first determined using time to approach and time to consume, and we clinical reference ranges for normal i-STAT blood parameters in the found that yogurt treats were highly valued and used for this study. common marmoset. Twelve rats were assigned to 1 of 3 treatment groups: surgery with meloxicam treatment, surgery with saline treatment, and anesthesia P118 Anaplastic Nephroblastoma with Peritoneal Metastasis in an only with meloxicam treatment. Rats were anesthetized with Adult Female Sprague Dawley Rat isoflurane and administered sustained-release meloxicam at 4 mg/kg or saline followed by a routine ovariectomy. Postoperative behaviors LA Quinn*, S Carrasco, S Muthupalani, JG Fox were assessed at 3, 6, 12, 24, and 48 h. This included consumption of yogurt treats, grooming, wound licking, rearing, orbital tightening, Department of Comparative Medicine, Massachusetts Institute of activity, ambulation, posture, and hair coat. While rats took the Technology, Cambridge , MA longest time to approach and consume the yogurt treats at 3 and 6 h postoperatively, there was no difference in the treatment groups. An 11-mo-old female Sprague Dawley breeder rat presented for Orbital tightening was the only behavior that was significantly necropsy due to an acute onset of lethargy, dyspnea, and persistent different at 3 h postoperatively, suggesting the surgical procedure abdominal distention 15 d postparturition. The 15 pups were active did elicit a pain response. These results suggest that the use of yogurt and nursing from the dam but were significantly undersized. The treat consumption as an indicator of pain is inadequate. pups were fostered to another dam in the colony whose litter was weaned the same day and were able to develop normally. On P120 Evaluation of Hoof Care Products for Duroc Swine (Sus scrofa physical examination, the dam was dyspneic and had a markedly domestica) Used in Biomedical Research distended, fluid-filled abdomen with a large, firm, multilobulated abdominal mass. Blood analysis indicated a stress leukogram, mild LL Mattox*1, D Mackessy1, N Burkey1, WA Bidot2, EJ Powers1, RA anemia, hypoalbuminemia, and evidence of dehydration. On Malbrue1 necropsy, the pleural and peritoneal cavities contained 5.0 and 13 mls of serosanguinous fluid, respectively. The right kidney was effaced 1University Laboratory Animal Resources, The Ohio State University, by a 2.0 x 3.0 x 3.0 cm pale-grey, multilobulated, firm to cystic mass Columbus, OH; 2University Of Florida, Gainesville, FL occupying the right retroperitoneal space. A second abdominal mass (3.0 x 2.0 x 1.5 cm) was covered by mesentery with multiple 0.4 - The hooves and pads of swine in a research setting are treated 0.8cm, pale-tan nodules. The nodules were adhered to the intestinal and maintained differently than that of those cared for in a farm serosa, pancreas, hepatic capsule, and ovarian bursa. Histologically, or production setting. Length and condition of toes along with the the right renal parenchyma was replaced by a poorly demarcated, health of the pads are 2 of the biggest concerns when housing swine. nonencapsulated, multilobulated, expansile mass composed of 3 Farm and research housing styles vary greatly, exposing the pigs neoplastic populations: epithelial, mesenchymal, and anaplastic to different types of flooring, moisture levels, and natural wearing blastemal-like cells. The epithelial population was composed of of the hooves. Excessive moisture can cause pads to become soft cuboidal to columnar cells arranged in acini and tubules surrounded and more susceptible to issues such as scratches, bruising, and by poorly differentiated neoplastic cells with 21 mitoses in 10 HPFs. cuts, making it uncomfortable for the pigs to walk. In our facility, Primitive glomeruli-like structures were occasionally present. The cases of Durocs presenting with cracked hooves and lameness were mesenchymal population was composed of spindle to stellate cells partially attributed to hoof maintenance, causing our institution to arranged in streams, expanded by an amphophilic, vacuolated investigate commercially available conditioning products and the stromal matrix. The blastemal population was composed of poorly frequency of use to help maintain moisture and the condition of differentiated polygonal cells arranged in nests exhibiting high hooves. For this project, we had a control group that did not receive nuclear to cytoplasmic ratio and 59 mitoses in 10 HPFs. The a hoof conditioning treatment and groups treated with 2 different extrarenal masses were predominantly composed of neoplastic commercially available hoof supplements. To measure effectiveness, mesenchymal and blastemal-like cells. Immunohistochemistry we used published scoring systems on locomotion and hoof lesions. revealed neoplastic epithelial cells were positive for pancytokeratin While both treatment groups showed marked improvement in the and mesenchymal and blastemal cells were positive for vimentin. condition of the hooves, data is still being evaluated to determine Neoplastic cells were negative for chromogranin A and S100. This which commercial product is most effective. The results of this case details a highly unusual, anaplastic nephroblastoma with project will aid in assessing possible methods to not only prevent, peritoneal metastasis. but also treat hoof concerns in swine, for production and research settings.

638 Abstracts of scientific papers 2019 AALAS National Meeting

P121 Development and Evaluation of a Novel Scoring System to identified. Review of health testing data showed that Proteus spp. Track Clinical Onset and Progression in Mouse Models of Prion (not on the MPF exclusion list) was detected in the production colony Disease location in Oct. 2015, suggesting that other nonexcluded organisms could also have gained entry. We replaced the MPF production M Beck* colony with 1 at the more restrictive excluded flora health standard in 2018, with all breeders sourced from the refreshed foundation Broad Institute, Cambridge, MA colony. Indeed this not only restored but improved the phenotype: the new production colony exhibited significantly earlier onset and Early signs of prion disease can be subtle and diverse, but severe much higher incidence compared to the old one. Our experience neurological defects and death follow swiftly thereafter. When using demonstrates that sensitive models require additional monitoring animal models to study prion disease, this complex onset necessitates and colony management actions to preserve and optimize desired strategies for early detection, and the rapid clinical decline requires model-specific phenotypes. Such considerations include genetic consistent methods of evaluation to ensure humane endpoints. To integrity and microbiome. address these needs, we formulated a behavioral scoring system to define and track clinical illness in mouse models of prion disease. P123 Development and Programmatic Application of a Low-cost According to this system, mice are regularly evaluated for common Actively Scavenged Anesthesia Nosecone for Mice signs of disease: visible tremor, kyphotic posture, 50% reduction in normal activity, generalized decrease in grooming, hind limb TR Meier1, KL Thompson1, J Nesbitt2, MJ Primitivo*1 weakness, periods of “staring,” and difficulty righting from lateral recumbency. During observation sessions, each mouse is evaluated 1Comparative Medicine, Mayo Clinic, Rochester, MN; 2Occupational for 15 sec. Observations are carried out weekly beginning at 90 d Safety, Mayo Clinic, Rochester, MN postintracerebroventricular inoculation with prion protein, increase to 3 times per week at 120 d postinoculation, and increase to twice Actively scavenged rodent anesthesia systems differ from passively per day upon disease onset, as defined by recognition of 2 or more scavenged anesthesia systems by the presence of a vacuum (negative signs of disease for 3 consecutive examinations. The experimental pressure) that facilitates pulling waste anesthetic gases away from endpoint is reached when an animal is displaying fulminant disease, the user and discharging them through either charcoal canisters or as defined by 5 or more signs of disease for 2 consecutive building exhaust. Passively scavenged rodent anesthesia systems are examinations. Among 175 mice evaluated, the average time to less effective than actively scavenged systems because the amount of disease onset was 148 d postinoculation, with animals reaching resistance to gas flow through the attached charcoal canister greatly fulminant disease in an average of 172 d. Typical disease progression exceeds the almost nonexistent resistance present in the gap between varied, but reduction in activity and visible tremor were identified as the animal’s face and the nosecone. For this reason, rodent anesthesia early indicators of disease, with a respective 51% and 41% of animals systems at our organization are required to use an active scavenging displaying these behavioral signs first. Fulminant disease was system for anesthetic waste gas management. There are commercially reached an average of 24 d subsequent to the identification of available mouse anesthesia nosecones that use active scavenging. reduction in activity, and an average of 17 d after onset of tremor. In These devices typically use a tube within a tube (coaxial) design. The contrast, kyphotic posture and “staring” were identified as a late- central tube delivers the gas anesthetic, and the outer tube is stage signs, with fulminant disease manifesting an average of 13 and connected to vacuum to scavenge any anesthetic gases that may 3 d after the observation of each respective sign. Using this system, escape around the nosecone-rodent interface. To help mitigate the we are able to accurately detect onset and identify clear clinical expense of the transition to actively scavenged nosecones, our endpoints in mouse models of prion disease. Furthermore, we are department designed and produced an active-scavenge mouse able to reliably predict the rapidity of clinical decline and the anesthesia nosecone using household tools and readily available necessity for proactive intervention. materials: 2 different sized brass tubes, a cold-weld compound, and 44 handgun brass casings. Our material cost per nosecone is less than P122 Optimization of Diabetic Phenotype in NOD Mice $2. The nosecone was evaluated by our safety office and determined to be as effective as commercially available versions. Due to our low MM MacBride*1, Z Li1, AV Perez1,2, PL Roesch1, SP Swing1, MP investment and ability to produce the devices in bulk, we elected to MacGowan1, K Hart1, DL Freer1, K Spencer1 provide them to laboratories at no cost. To date, we have built and distributed over 70 mouse anesthesia nosecones. This has helped 1Taconic Biosciences, Hudson, NY; 2Humodigen, Delmar, NY enhance and standardize our institutional safety profile for research projects using volatile anesthetics on rodents. The NOD strain develops spontaneous autoimmune type I diabetes. We monitor onset and prevalence of diabetes in our NOD colonies P124 Diarrhea and Associated Bacteremia during and following annually via urine glucose testing. The Murine Pathogen Free (MPF) Antibiotic Treatment in NOD-scid IL2rγnull Mice production colony showed a significant decrease in disease incidence over the period of 2014-2016; the gnotobiotic foundation colony M Stair*, S Carrasco, D Annamalai, N Fabian, A Mannion, S showed a significant delay in disease onset and decrease in disease Muthupalani, Y Feng, JG Fox incidence in 2015. Review of husbandry showed a few operational changes, but none were a likely root cause of the observed phenotype Division of Comparative Medicine, Massachusetts Institute of shift. Clinical data showed no inflammatory process or bacterial Technology, Cambridge, MA infection. Thus, we hypothesized that genetic drift due to continuous inbreeding and microbiome factors were more likely contributing Acute diarrhea and mortality were noted in a restricted access colony factors. We maintain a rigorous genetic quality program which of NOD-scid IL2rγnull (NSG) late gestation females and postparturient ensures that all inbred models undergo genetic refreshment every 5 y females with their litters. Neonatal mice were previously brought to or 10 generations of continuous breeding. By 2015, our foundation a shared procedure room for intracranial injections of human colony was refreshed as scheduled and gradually replaced the astrocytes. During a 4-mo interval, 27 mice were submitted for production colony during 2016-2017. Microbiome has been described complete necropsy, histopathology, and bacterial cultures and in the literature to affect NOD phenotype. Our foundation colony is antimicrobial sensitivity testing. Initial histologic examination of 7 maintained in a gnotobiotic isolator, which ensures that all microbial mice indicated 2 had neutrophilic enterotyphlocolitis and 5 had an organisms present are known, and entry of new organisms into the unusually heavy burden of Tritrichomonas spp in the ceca and isolator is rare. Our barrier production colony is maintained in an proximal colons. Aerobic cultures recovered Klebsiella environment that excludes specific microbial agents, while agents pneumoniae (Kp) from 1 blood and 4 cecal samples, and that are not specified on this exclusion list are not tested for or cytotoxic Escherichia coli pks+ from 2 blood and 4 cecal samples.

639 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

Antibiotic sensitivity of the E. coli pks+ indicated sensitivity to further evaluation by the veterinary staff. The primary differentials trimethoprim sulfa; given previous history of E. coli pks+ causing for the mass included infectious or neoplastic etiologies, and sepsis in mice, the drug was administered in the drinking water for 4 euthanasia was elected to obtain a definitive diagnosis. Gross wk. Twenty clinically infected mice were subsequently submitted, of necropsy revealed the mass to be associated with the scapula and which 55% (n=11) had histological lesions consistent with approximately 2cm x 1cm in size. Tissue blocks were submitted for suppurative bronchopneumonia, meningitis, enteritis, hepatitis, histopathology and immunohistochemistry staining. Hematoxylin metritis, and/or sepsis. Kp was cultured from pulmonary abscesses and eosin staining revealed a monomorphic population of and/or blood samples from 10 animals. Kp and E. coli pks+ were eosinophilic round cells suspect for lymphosarcoma. cultured from cecal contents of 12 and 7 mice, respectively. Kp was Immunohistochemistry staining using CD3 for T-cells and CD20 for commonly isolated in adult mice and significantly detected in ceca B cells was nonreactive, likely due to poor cross-reactivity between cocolonized with Tritrichomonasspp. Most Kp isolates (5/6) did not amphibian and mammalian antigens. Spontaneous neoplasms in exhibit a hyperviscous phenotype and were resistant to TMS (5/7). captive Xenopus spp are uncommonly described in the literature. Given Kp and E. coli pks+ isolates were susceptible to enrofloxacin, the Lymphosarcoma has previously been identified as the most common antibiotic was administered in the drinking water for 8 wk. This spontaneous neoplasm in laboratory Xenopus, however these tumors treatment resulted in a marked reduction of mortalities in the NSG more commonly arise from the liver, spleen, mesonephros, viscera, colony and the study successfully completed. Additional and thymus. This case highlights a common neoplasm in an preventative measures included changes to room entry and cage uncommon location. change order, increased use of disinfectants, personal protective equipment, and setting up a dedicated procedure space. Our findings P127 Pregnancy and Normal Infant Delivery in a Cynomolgus implicate that enteric Kp as the likely opportunistic pathogen causing Macacque (Macaca fascicularis) with a Bicornuate Uterus systemic infections in immunocompromised NSG mice and highlights the importance of antibiotic selection when treating dual ME Delehanty*1, G Hoffmann2, AE Baker3 enteric bacterial infections in NSG mice. 1Animal Welfare and Comparative Medicine, Covance, Oregon, P125 Urine Retention in a Rhesus Macaque (Macaca WI; 2Anatomic Pathology, Covance, Greenfield, IN; 3Early mulatta) following Sciatic Nerve Injury associated with Dystocia Development, Covance, Madison, WI

M Stovall*, DC Owens, M Crane, JR Johnston, S Gumber During a routine ultrasound to verify pregnancy in a mature female cynomolgus macaque (Macaca fascicularis) in our Development and Yerkes National Primate Research Center, Lawrenceville, GA Reproductive Toxicology colony, an anomaly was noted that appeared to be a possible bicornuate uterus. Ultrasonic evaluation A 13-yold female rhesus macaque (Macaca mulatta) presented with confirmed a pregnancy of approximately 19 d and an adjacent dystocia and hindlimb paralysis that was suspected to be due to structure that appeared to be a closed, nonpregnant uterine lumen. sciatic nerve impingement. A cesarean section was performed and The animal carried the pregnancy to term and delivered a normal the animal remained in cage housing for approximately 3 wk until female infant weighing 335.4 grams on gestational day 162 with no mobility improved. The animal was subsequently placed in an complications. After 6 mo, the maternal animal and young juvenile indoor enclosure within the social group for an additional 2 wk until were euthanized per protocol and evaluated at necropsy. The adequate mobility was obtained. During a routine annual physical maternal animal was found to have a bicornuate uterus with 2 exam approximately 3 mo later, the animal was noted with caudal distinct uterine horns sharing 1 uterine body and cervix. The abdominal organomegaly, which was suspected to be a subsequent maternal animal was also found to have unilateral renal agenesis, a pregnancy. The patient, however, failed to deliver an infant when sequela common to women with this congenital uterine abnormality. expected and was reexamined approximately 4 mo later. Caudal The female offspring was found to be normal anatomically at abdominal organomegaly was again noted on physical examination necropsy. Bicornuate uteri are considered to be relatively rare in and abdominal ultrasonography confirmed a markedly enlarged women and in cynomolgus macaques alike and can be associated bladder. The bladder was emptied via urinary catheterization, and with pregnancy loss. the animal began treatment for detrusor atony secondary to suspected sciatic nerve injury sustained during dystocia. Over the P128 Unresolved Atopy with Pruritus and Alopecia in a Rhesus next few months, the animal was frequently treated for urinary Macaque (Macaca mulatta) retention and urinary tract infections. The condition continued to worsen leading to vaginitis and abdominal distention, so euthanasia ML Martin*, M Hanson was elected. Histological evaluation revealed bilateral sciatic nerve and cauda equina degeneration, necrotizing serositis of the cervix/ Veterinary Service Program, Walter Reed Army Institute Research, vagina, and a mild cystitis. Prolonged labor has been identified as a Silver Spring, MD risk factor for urinary retention in humans, and sciatic nerve degeneration has been noted in cattle following dystocia. For An 8-y-old, 8.32-kg male rhesus macaque (Macaca mulatta), single nonhuman primate cases of dystocia, the possibility of sciatic nerve housed in an indoor, climate-controlled animal facility presented degeneration and subsequent urinary retention should be considered. with a 3-y history of alopecia and pruritus that appeared to worsen over the past 12 mo. Previous treatment include omega 3 fatty acids P126 Periscapular Lymphosarcoma in a Xenopus laevis and vitamin E supplementation, with no improvement. Physical exam identified severe alopecia over the head, limbs, abdomen, flank, MT Lieberman*1, H Warren1, S Muthupalani2 and back, with erythematous, thickened, and flakey skin. The affected areas appear extremely pruritic with multifocal self-inflicted 1Harvard Center for Comparative Medicine, Harvard Medical abrasions. Skin scrapings were negative for external parasites. School, Boston, MA; 2Division of Comparative Medicine, Biopsies taken 6 mo apart from 9 different locations revealed mild to Massachusetts Institute of Technology, Cambridge, MA moderate, superficial perivascular lymphoplasmacytic inflammation. Serum chemistry and complete blood count (CBC) performed every 6 A 5-y-old female Xenopus laevis presented with a 1cm x 1cm mo was unremarkable. A thyroid panel was also normal. Blood was subcutaneous mass in the left cervicoscapular area. The frog had taken and submit to ACTT for food allergy testing, all positives and been used for a single nonsurgical oocyte collection between July and border line positives were removed with no effect. Oclacitinib (0.4 August 2018, and had no clinical signs of illness. Two years prior to mg/kg) was administrated for 1 mo with no improvement. The dose presentation, a different frog in the colony was noted to have a was then increased to 0.8 mg/kg for 2 wk with a pharmacokinetic similar subcutaneous mass, but was euthanized by the lab without analysis to verify a therapeutic dose had been achieved. Although

640 Abstracts of scientific papers 2019 AALAS National Meeting

there was an increase in new hair growth, there was no improvement body weight peaks differed by sex (male 14M , female 22M ) in B6J in clinical pruritus. Oral medication was stopped and a topical mice. Food/ water consumption increased rapidly at over 22 mo in treatment of essential oils was implemented for 5 mo with minimal both B6 mice. Survival rates started to decline at approximately 18 improvement in pruritus and no new hair regrowth. A CBC and mo and were somewhat lower in female aged mice (B6N). Rotarod serum chemistry was performed and revealed mild eosinophilia, performanced peaks at 3 mo in male and at 6 mo in female B6N mice. neutrophilia, and a monocytosis. In addition to topical treatment, Rotarod performanced peaks as well at 3 mo in B6J mice, and then oclacitinib was restarted at 0.8 mg/kg. Ideally, the next step in continues to decline in both strains. Rotarod performance and grip diagnostics would include intradermal allergy testing, and/or strength of aged animals showed relatively low score. Enlarged elimination diets. However, extent of diagnostics and treatment given seminal vesicles or splenic tumors were often found at autopsy of to a laboratory animal with atopic-like dermatitis should be carefully both strains. Urinary corticosterone levels were relatively higher in determined by considering the health and wellbeing of the animal female mice and tended to increase with age in both B6 mice. Blood and the potential inability to use on protocols due to underlying tests showed that the total WBC count started to decline at around 18 chronic inflammation. Currently, this animal is being used as a mo and the composition of WBC tended to change with aging. training animal for PI familiarization and non-invasive procedures. Various age-related changes (such as body weight, motor performance, the total WBC count, and its composition) found in our P129 Three Alcohol-based Agents Used for Presurgical Skin aged B6N/B6J mice could be candidate senescence indicators at Preparation in Mice individual level. We will investigate these parameters in detail and continue to search for novel biomarkers (such as fecal IgA levels). M Huss*1, K Casey1, J Hu1, R Moorhead1, H Chum2 P131 Eradication of Astrovirus by Crossfostering 1Comparative Medicine, Stanford University, Stanford, CA; 2Palo Alto Veterans Affairs, Palo Alto, CA P Sharp, I Faseeh*

Proper aseptic technique is a crucial component of rodent survival Animal Resources Centre, Murdoch, Australia surgery. Ease of technique, space constraint, batch surgery, and cost Astrovirus is a single stranded, unenveloped RNA virus that was are factors that may affect researcher compliance with proper aseptic orginally discovered in the mid 1970s associated with diarrhea in technique. This study evaluated 3 different antiseptic preparation humans. Since its discovery it has been found both in birds and agents that simplify application and/or cost compared to standard mammals, including both wild and laboratory rodents. Frequently, triplicate application of povidone-iodine and alcohol. Euthanized immunocompromised mice are affected and it has been determined mice (n=40) were shaved on the dorsum and culture swabs were that the adaptive immune system plays an essential role in restricting taken for RODAC plating and bacterial identification. Shaved sites viral replication. While the clinical or research significance of the were prepared using 1 of the 4 antiseptic preparation agents. virus is unknown, we describe its eradication in laboratory mice Cultures were again taken for evaluation immediately following, and through crossfostering. Crossfostering has been used to successfully 20 min after antiseptic preparation. In the second part of the study, eradicate Helicobacter spp, murine norovirus, mouse hepatitis virus, eight mice (n=2 per group) were prepared with 1 of the 4 antiseptic and mouse encephalomyelitis virus in laboraotry mice. The colony preparation agents for a survival surgical procedure in order to has been astrovirus free for over a 1 based on PCR-based assessment. evaluate if the antiseptic preparation agents caused skin irritation or impaired healing. Results from this study indicated that the 3 P132 Use of Neurosonography to Monitor Treatment of a Cerebral antiseptic preparation agents were all equally effective at reducing Abscess in a Rhesus Macaque (Macaca mulatta) bacterial populations at the immediate and 20-min postpreparation timepoints. Histopathological examination of the incision sites PC LaTourette II*1,2, GK Adams3, K Sharika3, CK Wallace1 revealed normal signs of healing and did not result in histologic lesions adjacent to the incision site. We conclude that all 3 products 1University of Laboratory Resources , University of Pennsylvania, are comparable to povidone-iodine and alcohol preparations as Philadelphia, PA; 2Pathobiology, University of Pennsylvania, agents in the aseptic preparation of surgical sites. Philadelphia, PA; 3Neuroscience, University of Pennsylvania, Philadelphia, PA P130 Search for Characteristic Change of the Strain and Definitive Senescence Biomarkers in Naturally Aged C57BL/6 Mice A 15-y-old male rhesus macaque (Macaca mulatta) on a neuroscience protocol, and 1-wk post cranial implant revision, presented with N Ogiso*1, S Matsubara1, A Julio1, T Sazi1, K Tomita2, K Yamaguchi2, acute lethargy and anorexia. Clinical signs progressed to dull T Shinya2, M Maruyama3 mentation, trembling, ataxia, and head pressing. Blood work did not reveal any abnormalities. CSF cytology revealed an eosinophilic 1Laboratory of Research Animal, National Center for Geriatrics and pleocytosis with degenerative changes, and a bacterial culture of the Gerontology, Obu-city, Aichi, Japan; 2KAC Corporation, Kyoto, implant margin grew Staphylococcus pseudintermedius. An MRI Japan; 3Mechanism of Aging, National Center for Geriatrics and revealed 2 areas just deep to the chamber with hypoattenuated Geron, Obu-city, Japan centers circumscribed by hyperattenuated boundaries measuring about 0.5 cm in diameter each, diagnosed as cerebral abscesses. A Our facility has kept many naturally aged animals (mice and rats) reactive encephalitis and meningitis were present, likely secondary to used for gerontology and geriatric researches. If a scientist conducts the abscessation. Systemic antibiotic therapy was initiated research using these animals, it is important to consider the effects of (ceftriaxone 50 mg/kg SID IM for 8 wk). Since weekly MRIs were various genetic and environmental factors. However, a clear criterion impractical and the abscesses were located beneath the craniotomy for defining aged mice has not been established. We report various chamber, response to treatment was monitored via ultrasound. Serial strain difference and age-related characteristics observed in our mice. measurements were obtained using an obstetrical transvaginal probe Male and female C57BL/6 (C57BL/6NCrSlc(B6N), C57BL/6J(B6J)) directed through the chamber overlying the craniotomy site while the mice (4-wk-old) were purchased every 3 mo and kept over their animal was awake and restrained in a primate chair. Weekly lifetime. Physiological (measurement of body weight, food/water neurosonography showed a decrease in abscess diameter and consumption and survival rates), behavioral (the rotarod tests and dissolution of the surrounding abscess capsule. The ultrasound the grip strength tests, morphological (autopsy and histological sessions were augmented with MRI that confirmed resolution of the examination), biochemical (urinary corticosterone), and lesions and associated encephalitis and meningitis. hematological analyses were performed. Body weight peaks at 16-18 Neurosonography has previously been described for the mo-old (M) in B6N mice (approximately male 44.4g, female 33.8g) experimental use of neural recording probes in vivo. This abstract is and only male mice showed rapid decrease at around 22 mo. Also the first to describe transchamber neurosonography for clinical

641 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 imaging in a nonhuman primate. In addition, this technique has been epicardium of 2 of 16 mice. Multifocal myocardial mineralization and used in our department for monitoring bone regrowth after cranial cardiomyocyte degeneration were identified in 12 of 16 mice. implant removal in awake rhesus macaques. Neurosonography could Additional findings included skeletal myodegeneration, necrosis and be expanded clinically for diagnosing and monitoring mineralization (6/16), a spinal cord infarct (1/16), edema affecting encephalopathies in chronically implanted animals with lesions multiple organs (3/16), hepatitis (2/16), typhlitis and colitis (3/16), directly underneath or very close to the craniotomy site. and graft versus host disease (2/16). Potential differential diagnoses for the myocardial and skeletal muscle lesions include toxicity, renal P133 Evaluation of the Incidence and Severity of Pododermatitis in disease , paraneoplastic syndrome, endocrine disease Rats Housed Longterm Singly or in Pairs with Square or Round (hyperparathyroidism), and nutritional deficiency. These were Polycarbonate Shelters considered unlikely based on the appearance and distribution of lesions, normocalcemia in all affected mice, and the clinical and PM Gerwin*2, ER Berryman2, C Halsey1 experimental history. Strain related myocardial mineralization has been described in C3H and DBA mice but not in NSGs. This case 1Global Pathology, Pfizer Worldwide Research & Development, series may represent a newly recognized strain related background Groton, CT; 2Comparative Medicine, Pfizer Worldwide Research and lesion. Development, Groton, CT P135 Serum Biochemical Changes during Pregnancy in Common Pododermatitis, defined as inflammation of the skin of the paw, has Marmosets (Callithrix jacchus) been associated with rats housed in wire bottom cages and less commonly associated with rats housed on bedding in solid bottom RM Kramer*1, SC Artim1, MA Burns1, CH Toolan1,2, A Sheh1, JG Fox1 cages. Other factors that may contribute to the development of this condition include high body weights, stock/strain, longterm 1Division of Comparative Medicine, Massachusetts Institute of housing, and housing on coarse or wet bedding. Few treatments have Technology, Cambridge, MA; 2College of Veterinary Medicine, been shown to be effective and the longterm prognosis remains University of Pennsylvania, Philadelphia, PA guarded. A preliminary assessment of rats (n=169) housed on solid Profound physiologic changes occur during pregnancy, many of bottom cages with alpha cellulose bedding for 6 mo showed 44% of which lead to changes in serum biochemical parameters. These are single (n=80) and 36% (n=89) of pair-housed rats developed hind foot well documented in humans and reported in several species of lesions. Lesions were more severe in single-housed rats. A nonhuman primates, including Rhesus macaques (Macaca mulatta), subsequent study was conducted using 71 male Wistar Han rats African green monkeys (Chlorocebus aethiops), squirrel monkeys (Crl:WI[HAN]) maintained for 6 mo for safety pharmacology studies. (Saimiri sciureus), and baboons (Papio spp.). Such changes have not Rats were housed singly (n=35) or in pairs (n=36) in open top cages been previously evaluated in the common marmoset (Callithrix and, in addition to manipulanda, were housed with 1 of 3 shelter jacchus). Marmoset breeding and medical records from pregnant and items: a round polycarbonate tunnel, a square polycarbonate tunnel, nonpregnant females were retrospectively examined, and reference or paper nesting material with no polycarbonate tunnel. We ranges for blood parameters during pregnancy in healthy animals hypothesized that an increase in number and severity of foot lesions were determined using 40 serum chemistry panels from 16 different would occur in animals housed singly and in animals housed with females. Values for pregnant and nonpregnant healthy animals were square shaped shelters. Body weights were taken monthly and foot compared using a student’s t test (nonpregnant n=148 chemistry lesions were scored monthly on a 0-3 scale (no lesion, mild, panels from 44 females). There were significant decreases in serum moderate, severe). Dynamic weight bearing testing was performed at albumin (P = 0.007), total calcium levels (P = 0.007), phosphorus (P = 0, 3, and 6 mo. Additionally, histopathology will be reviewed. At 5 0.004), and cholesterol (P = 0.001) in pregnant animals, as in other mo, 17% of single- and 3% of pair-housed rats developed foot lesions. nonhuman primates. Changes in serum ALP and BUN levels did not For single-housed rats, 41% of animals housed with square shelters change significantly in marmosets, though these parameters are developed lesions compared to 8% with round shelters and 17% with affected by pregnancy in some species. Since serum chemistry panels paper nesting material. Only single housed animals had lesions on are routinely used for health monitoring and clinical decisionmaking, more than 1 hind foot and had lesions that were scored as moderate. including as a screening tool for several common diseases in Our preliminary data support our hypothesis that single-housed marmosets, knowledge of how a normal pregnancy affects blood animals and animals in square-shaped shelters develop more lesions chemistry parameters is important for interpretation of clinical and those lesions tend to be more severe. These new data will allow pathology data in pregnant animals and management of marmoset us to provide housing conditions and enrichment that may help breeding colonies. reduce the incidence and severity of pododermatitis in rats. P136 A Comparison of Suture and Metallic Clip Incisional Closure P134 Morbidity and Mortality associated with Cardiac Methods in Rattus norvegicus Mineralization in NSG Mice J Schaphorst*, T Beltran, S Hegge RJ Floyd*1, AO Michel1,2, A Piersigilli1,2, K Lertpiriyapong1,2 US Army, Tacoma, WA 1Tri-Institutional Training Program in Laboratory Animal Medicine and Science, New York City, NY; 2CCMP, Center of Comparative During study iterations involving a rat model for creating abdominal Medicine and Pathology, New York City, NY adhesions via laparotomy procedure, we found that 25% of rats (n=69) self-removed their incisional metal clips (“staples”) Over 8 mo, 24 of 290, 2-to 5-mo-old NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ postoperatively. This necessitated repeat anesthesia and replacement (NSG) mice from a single vendor, used by several laboratories, of the clips, and introduced the potential for complications such as presented with low body condition scores, hunched posture, pallor, pain, infection, or evisceration. This self-removal did not improve lethargy, increased respiratory rate and effort, and distended despite frequent monitoring, provision of pain medication, nor with abdomens. All mice were maintained in individually ventilated cages the use of protective jackets. Our clinical hypothesis was that perhaps with autoclaved aspen chip bedding, irradiated feed, and acidified the rats were irritated or annoyed by the somewhat bulky presence of RO water. Complete necropsies were performed on 16 of the affected the metal clips and thus self-removing them by chewing or other mice. Fifteen mice had been implanted with human xenografts and 1 manipulation, and perhaps an alternative closure technique would was naïve. Complete blood cell counts and serum chemistry reduce the incidence of reopening incisions by the rat. A comparison performed on 6 of 16 mice revealed a mild regenerative (2/6) or of staple versus suture incisional closure was conducted. In the next nonregenerative anemia (2/6) and an elevated AST (5/6). On gross scheduled study iteration, 10 rats had their laparotomy incision sites examination, disseminated, white, pinpoint foci were seen on the closed using skin sutures (n=5) or staples (n=5). As with previous

642 Abstracts of scientific papers 2019 AALAS National Meeting

iterations, rats were provided local anesthesia intraoperatively, long- lab verified in-house results and identified severe elevation of acting pain medication, and postsurgical daily monitoring. Of the creatine kinase (CK), AST, ALT, and phosphorus which were two groups, 1 rat in the staple group self-removed the day after suggestive of rhabdomyolysis and acute renal injury. Fluid therapy, surgery, but no rats in the suture group removed their skin sutures diuresis, and correction of metabolic disturbances were initiated. throughout the monitoring period. At 14 d rats were euthanized as Once stabilized, a thorough assessment of the anesthetic records part of the adhesion study endpoint, and tissues from the laparotomy revealed a late onset, creeping hypercapnia, tachycardia, and incision sites were collected. Also, no differences in postsurgical hyperthermia which were unresponsive to hyperventilation, appearance, behavior, or posteuthanasia incision area histopathology isoflurane manipulations, and temperature adjustments. A were observed between the 2 groups. Although the study sample size presumptive diagnosis of malignant hyperthermia (MH) was made of 5 rats per group is too small to draw statistical significance, the based on this history and the clinical findings. Over the course of the method of suture closure was deemed preferable to metallic clips by next 5 d, fluid therapy with adjustment of electrolyte and blood pH laboratory staff. Because method of incisional closure does not affect was performed, however, the gold standard intervention for the outcome of the primary study, and also represents a refinement malignant hyperthermia, dantrolene, was not available for use. technique, we recommend using this closure method for rats Throughout treatment, uremia, blood electrolyte and pH, and urine undergoing laparotomies at our facility. output were monitored daily in-house and supplemented with periodic full biochemistry panels. Uremia peaked between 48 and P137 Iatrogenic Reovirus Infection in Highly Immunocompromised 120 h but returned to normal by 30 d postoperative. Urine output Mice was nil during the first 60 h postoperation but was reestablished following furosemide treatment and rose from oliguric levels to SJ Mangosing*1, RJ Ricart Arbona2, S Monette1,2, N Lipman1,2 normal levels by 10 d postoperation at which time the animal was medically cleared. Given the genetic origin of MH, whole genome 1Tri-Institutional Training Program in Laboratory Animal Medicine sequencing was performed using single molecule, real-time and Science, New York, NY; 2Center of Comparative Medicine and sequencing technology on samples collected from the patient and a Pathology, Memorial Sloan Kettering & Weill Cornell Medicine, New healthy control animal to screen for mutations in genes commonly York, NY involved in MH. To the authors’ knowledge, this is the first successfully managed case of MH in a macaque and genetic testing Reovirus infections are rare in contemporary mouse colonies. Natural may yield useful data to exclude animals with MH mutations from infections typically occur in young and immunosuppressed mice. studies requiring inhalant anesthetics. Infection is typically pantropic with virus recovered from the liver, brain, heart, pancreas, spleen, lymph nodes, and blood vessels. P139 A Syndrome of Duodenal Ulceration with Strictures in a Lesions include hepatic, neuronal, pancreatic, and myocardial Colony of Common Marmosets (Callithrix jacchus) necrosis; pulmonary hemorrhage; hepatitis; and, encephalitis. We observed 4 distinct occurrences of reovirus infection in adult NOD. SC Artim, MA Burns*, A Sheh, JG Fox, S Muthupalani Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice engrafted with patient-derived tumor xenografts (PDX) over a 3-y period. Two of these outbreaks Division of Comparative Medicine, Massachusetts Institute of were detected as a result of reovirus seroreactivity in soiled bedding Technology, Cambridge, MA sentinel mice, whereas in one, clinical signs consisting of tail necrosis, unexpected early morbidity and mortality, were observed. Gastrointestinal diseases including an IBD-like syndrome are Representative mice from all four of these outbreaks (n=15) were endemic in captive common marmoset colonies. We describe a novel subject to complete necropsy. Eight mice had a subcutaneous syndrome characterized by duodenal mucosal ulceration and xenograft. Microscopic lesions included hepatitis and/or hepatic stricture in 21 marmosets. The syndrome occurred predominantly in necrosis (12/15), adrenocortical necrosis (11/15), interstitial younger animals; 15/21 were 2-3 y of age at time of necropsy. The pneumonia (11/15), as well as ovarian (5/15) and myocardial (5/15) syndrome affected both sexes (13 females, 8 males) and 15/21 cases necrosis. The 3 mice with tail lesions had ischemic necrosis caused by had at least 1 affected sibling. All but 1 of the animals were derived vascular thrombosis. In situhybridization (ISH) using a reovirus- from the same source colony. Clinical signs included vomiting specific probe was performed in 6 of these mice and revealed co- (17/21), diarrhea (18/21), and weight loss or poor weight gain localization of viral nucleic acids with lesions in the adrenal cortex, (21/21). A key finding was a palpable mid-cranial abdominal ovary, lungs, heart and liver. Feces from 5/15 of the cases tested organomegaly, thickening or mass (19/21). In select cases, ultrasound positive for reovirus by PCR. These cases highlight unusual lesions, imaging revealed evidence consistent with duodenal mucosal specifically adrenal, tail, and ovarian necrosis. Three tumor seeds ulceration. Blood chemistry and complete blood count analysis transplanted into affected mice tested positive for Reovirus by PCR, revealed hypoalbuminemia, hypoglobulinemia, hypoproteinemia, however we have not yet identified how those tumor seeds became hypocalcemia (total), elevated alkaline phosphatase, anemia, and in contaminated. Outbreak history, clinical and pathologic findings, and some cases leukocytosis. Animals were managed with supportive the potential sources of the virus will be presented. care, antiulcer therapy, antimicrobial therapy, and budesonide. On necropsy, consistent gross findings were stricture and/or visible P138 Management of Acute Kidney Injury Subsequent to ulceration at 0.5-1 cm near the major duodenal papilla. Distended Postoperative Rhabdomyolysis in a Rhesus Macaque (Macaca gall bladder and cystic ducts along with variable dilatation of the mulatta) proximal duodenum and stomach were also observed. Mesenteric lymphadenopathy and mesenteric-pancreatic adhesions were also SC Adams*1,2, K Williams2, K Berry1, C Guyot1, M Leblanc1 frequently noted with intestinal perforation in a few cases. Main histological findings included duodenal mucosal ulcerations with 1Animal Resources Department, Salk Institute, La Jolla, CA; 2Salk associated chronic-active granulocytic and lympho-histiocytic Institute for Biological Studies, La Jolla, CA inflammation with variable fibrosis and reactive epithelial proliferation at the affected stricture sites. Other related findings A 6-y-old male rhesus macaque (Macaca mulatta) presented for were variable grades of subacute to chronic fibrosing cholecystitis obtundation 1 day postsurgical placement of a head post for use in a and choledochitis along with chronic-active pancreatitis secondary to visual neuroscience study. On examination, the animal evinced pale the ulcerated duodenum. No infectious etiology was identified based mucous membranes, a CRT over 4 sec, hypothermia, and on bacterial culture, histopathology, and electron microscopy of gall dehydration. Initial in-house clinical pathology confirmed severe bladder and duodenum. This novel duodenal mucosal ulceration and dehydration as evidenced by hyperproteinemia and elevated PCV, as stricture syndrome in a young adult marmoset cohort is being further well as metabolic acidosis, hyponatremia, hyperkalemia, investigated to understand its etio-pathogenesis and to aid future hypochloremia, and uremia. A full biochemistry panel at a reference clinical management strategies.

643 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P140 Eye Spy: Ocular Opacity in a Guinea Pig (Cavia porcellus) P142 Assessment of Rodent Tail Tattoo Ink for Microbial and Chemical Contamination SA Kurnick*, JL Haupt, S Muthupalani TR Young*1,2, T Whiteside2, J Locklear2 Division of Comparative Medicine, Massachusetts Institute of Technology, Cambridge, MA 1CMB, Alpha Omega/NIEHS, Durham, NC; 2Quality Assurance Laboratory, Durham, NC A 4-y-old experimentally naive female intact Hartley guinea pig presented with a white opacity at the limbus of the right eye. At 922g, Rodents are the most frequently used animal models in the biological the animal had a BCS of 3+/5 with no other abnormalities noted on research field. Unique animal identifiers are often needed, especially physical examination. The other 3 animals in the colony were for breeding and experimental studies. Tail tattooing is a common unaffected. Differential diagnoses included trauma (corneal scarring), method for identifying mice. Tail tattoos have been widely used for metabolic (ocular lipidosis), or neoplastic/nutritional causes decades as a reliable and clear method for rodent identification. (lymphosarcoma, hypervitaminosis D). An ophthalmic exam However, this method can result in skin inflammation and primary localized the lesion to the anterior chamber using direct or secondary skin infections. The purpose of this study was to survey ophthalmoscopy and slit lamp biomicroscopy. There was no evidence our tattoo inks and equipment for microbial and chemical of discharge, squinting, pain, or fluorescein stain uptake. Both eyes contamination. Bacterial isolates were identified using traditional retropulsed normally. Tonometry examination was unavailable. microbial techniques and confirmed with matrixa-assisted laser Serum chemistry was within normal limits, whereas urine chemistry desorption ionization-time of flight mass spectrometry (MALDI-TOF suggested elevated protein excretion and calcium to phosphorus MS) and 16S PCR/sequencing. The tattoo inks (black, blue, and red) ratio. No abnormalities were visible on lateral, dorsoventral, or were analyzed for heavy metals contamination using pressure ventrodorsal radiography of the skull. In the absence of discomfort, digestion and inductively coupled plasma mass spectrometry (ICP- the guinea pig was monitored for the following 6 mo with no visible MS). The most common bacteria isolated from the tattoo components change to the opacity until the animal was euthanized for unrelated were Enterococcus casseliflavus, Corynebacterium lubricantis, Bacillus reasons. A diagnostic necropsy revealed bilateral ovarian cysts and niacin, Bacillus simplex, Lactobacillus murinus, Lysinibacillus adrenal hypertrophy. Following postmortem enucleation, attempted boronitolerans or macrolides, and Acinetobacter radioresistens. Our study aspiration of the anterior chamber deposit was unsuccessful due to demonstrates that tattoo inks contain microbial contaminants that the rigid nature of the lesion. On histopathology, the ocular changes may initiate or contribute to inflammation and infection at the tattoo were consistent with bilateral heterotopic bone formation of the site. The tattoo inks also contain detectable levels of heavy metals anterior uvea. While guinea pigs are prone to metastatic or (arsenic, cadmium, lead, and mercury). Both of these contaminants of dystrophic calcification, heterotopic bone formation is a unique tattoo ink should be considered in study design. clinical entity characterized by the presence of functional bone marrow. This lesion has been reported in aging pet guinea pigs and P143 Acute Pruritus in a Juvenile Yucatan Minipig (Sus scrofa should be a diagnostic consideration regarding ocular abnormalities domestica) in laboratory-maintained guinea pigs. W Smallridge*2, AK Brice2,1, B Philips1 P141 Identification and Treatment of the Common Snake Mite Ophionyssus natricis in Research Pythons 1University Laboratory Animal Resources, University of Pennsylvania, Philadelphia, PA; 2School of Veterinary Medicine, TT Mufford*, JD Reuter Department of Pathobiology, University of Pennsylvania, Philadelphia, PA Office of Animal Resources, University of Colorado Boulder, Boulder, CO A 5-mo-old, 31-kg male castrated naïve Yucatan (Sus scrofa domestica) housed in an indoor research facility presented to the clinical service Snakes are uncommonly used as models in biomedical research and for decreased activity and appetite 2 wk following release from therefore SPF purpose-bred animals are in short supply. Laboratories quarantine. No abnormalities were noted on physical examination, often rely on exotic pet breeders and individual small breeding and the animal was started on a gastroprotectant regimen of colonies as sources for colony animals. These breeders, while usually sucralfate and famotidine to treat presumptive gastric ulcers. Three ethical and reputable, do not generally have a preventative health days after initiation of treatment the animal was reported for care program as stringent as most contemporary vivaria. A university erythema, crusts, and hyperkeratosis associated with the base of the houses a Burmese and Ball python colony which had traditionally right ear. The following day the lesions had extended to the inguinal, been managed by investigator staff. The Office of Animal Resources abdominal, and axillary regions, and the animal was extremely partnered with the laboratory to share resources and enhance colony pruritic. Initial diagnostics included superficial skin scrape, cytology, management. Laboratory staff detected Ophionyssus natricis, the bacterial and fungal cultures, and bloodwork, all of which were common snake mite, as evidenced by live mites on paper liners and unremarkable. The animal was prescribed ivermectin and cephalexin on animals approximately 2 wk after they were housed into the as presumptive treatment for ectoparasites or bacterial infection. vivarium. This was reported to veterinary staff and confirmed via Despite this, lesions continued to progress, with the animal microscopic evaluation. The vendor was contacted and reported a developing hyperkeratosis and vesicular dermal lesions along the pyrethrin soak as treatment prior to shipment, which had failed. dorsum. The animal was sedated for collection of skin biopsies and Upon review of the literature veterinary staff initiated topical blood for scabies ELISA. Scabies ELISA was negative, and on ivermectin treatment for the mites, which also failed. After extensive microscopic examination of the lesions, there was a moderate, research and consultation with a local exotics veterinarian, we diffuse, eosinophilic, and histiocytic infiltrate within the superficial initiated a new treatment cycle using a commercially available dermis and surrounding blood vessels throughout the dermis, fipronil spray and intensive thorough housing sanitation regimen. admixed with fewer lymphocytes. Multiple to extensive intracorneal With the help of the facility manger, research laboratory, veterinary, and intraepidermal pustules and moderate, diffuse orthokeratotic and husbandry staff all snakes were successfully treated and the hyperkeratosis were also observed. The primary differential mites were eradicated. With new shipments of snakes arriving from diagnosis was severe allergic skin disease caused by food allergy, various sources, we revised our original SOPs, husbandry and cage environmental allergies, or hypersensitivity to ectoparasites. Oral wash procedures, and veterinary care to assure safe handling and medications and treats were discontinued, and the animal was avoid accidental fomite transmission and maintained a mite free transitioned back to vendor feed. Lesions steadily improved and python colony. Nontraditional species often present challenges that were nearly resolved at the time of euthanasia and tissue collection 3 require creative strategies, diligent observation, and a robust support wk after presentation. While definitive diagnosis is not possible team to sustain healthy animals. 644 Abstracts of scientific papers 2019 AALAS National Meeting

without performing a food challenge, we suspect that this animal’s (n=9 cages) or conjoined cage configurations of up to 2 cages (n=10 dermatitis may have been secondary to a food allergy. This case cages), on the breeding efficiency of C57Bl/6 mice, as compared to an highlights the importance of a complete diagnostic work-up when individually ventilated shoebox cage system (n=9 cages). Mice were evaluating skin disease in swine, as conditions like allergic skin housed in breeding groups ranging from a 1:1 male:female breeding disease can closely resemble infectious dermatoses in this species. pair in a single cage, up to 2:8 male:female breeding groups in conjoined cages. Efficiency metrics include litters per breeding group P145 Breeding and Colony Management Assistant: A Tool or female over time, litter survival, weaning rate, pups weaned by Designed to Maintain Minimally Inbred Standardized Complex number of females per cage, and weight trends from birth to Microbiota Rodent Colonies weaning. Overall, caging treatment, caging type, or number of females per cage had no statistically significant effect on the A Sharma*, AL O’Neill, A Dewitt, M McDowell, J Waterman, CL percentage survival of pups weaned. Conjoined cages produced Franklin fewer litters per female over the examined time period as compared to either singular cage types. Pie-shaped animal systems can Veterinary Pathobiology, University of Missouri, Columbia, MO maximize available spacing in research settings, and while neither pie-shaped configuration (single or conjoined) had a negative effect Understanding the causal relationship between the composition of on pup survival and weight gain, they do not appear to improve gut microbiota (GM) and disease model phenotype is a rapidly breeding efficiency. emerging challenge for biomedical researchers. To facilitate such investigations, we recently developed series of mouse colonies that P147 Successful Group Housing of Female Laboratory Rats after possess differing standardized intestinal microbiota that represent Head-cap Implantation the microbial spectrum of that seen in contemporary rodent colonies. These mice can be used as embryo transfer surrogates for derivation A Robinson* of any mouse model of disease that possess one or more desired GM profiles. We have shown that composition of the resident GM is a IVSD, GlaxoSmithKline, Ware, United Kingdom primary determinant of disease severity in models ranging from inflammatory bowel disease to autism and provided proof-of-concept It’s well accepted that social housing of rats is important for their that this approach can be used to investigate the contribution of wellbeing. However, after head-cap preparation, rats are generally complex GM on disease phenotype and model reproducibility. To individually housed to reduce the risk of damage caused by chewing. optimize assisted reproductive and maternal characteristics, these In 2016 we had a request in for a new model that required head-caps. colonies were created to form genetically diverse (i.e. outbred) mice We wanted to refine the housing conditions for the animals by group and are maintained using a minimal inbreeding strategy. Moreover, housing postsurgery. To facilitate this, we discussed the idea with the breeding and maintenance requires collaboration with multiple named veterinary surgeon (NVS) and surgery team to find solutions laboratories and generates abundant data that must be shared among that would enable us to achieve our goals. We agreed to regroup the these labs. Hence, we developed a user-friendly macro-enabled rats postsurgery in trios and monitor closely for signs of chewing. visual basic application (VBA) Excel interface termed as Breeding Initially there was no damage to the head-cap but the dust-caps and Colony Management Assistant (BCMA). Powered by >100 (which protect the head-cap) were chewed off and needed to be macros and VBA codes, BCMA has powerful analytical and search replaced daily, and the rats hit their heads on the low gridded lid. We capabilities facilitating day-to-day tasks such as data entry, records wanted to keep them in social groups because as soon as they had maintenance, and meeting the needs for breeding, weaning and recovered they interacted with each other, and housing in taller cages genetic refreshment. The BCMA was developed keeping in mind the avoided trauma to their dust-caps caused by the cage lid. After needs of technicians, animal research staff and principal further discussions we decided to house them in groups of 4 for investigators. Overall, the BCMA has proven to be highly flexible and approximately 100 d in our large tower cages. Enrichment included suitable for diverse management needs including: a) maintenance of suspended tunnel, nesting, and a chew block. We also redesigned the minimally inbred mouse colonies, b) minimization of animal dust-caps to make them lighter, stronger, and larger. After making numbers while meeting specific age and sex requirements for lab these changes the rats were unable to chew the dust-caps off and it experiments and services, and c) an annual genetic and microbial reduced the number of times staff had to handle the dust-cap. refresh. Housing the rats in larger cages prevented the dust-caps being knocked and increased the amount of times the technicians noticed P146 The Effect of “Pie-Shaped” Caging Systems on Breeding the rats huddled together compared with rats in the standard cages. Efficiency All animal studies were ethically reviewed and carried out in accordance with the corporate policy on the care, welfare, and A Winn*1, C Thurman1, L Paluch1, M Klinger1, G Roble1,2 treatment of animals.

1Office of Veterinary Resources, New York University, New York, P148 Effect of Commercially Available Bedding Substrates on NY; 2Fred Hutch Cancer Research Center, Seattle, WA Mouse Gut Microbiome

Research facilities often face housing constraints as rodent colonies AF Hoggatt*1, C Hallman3, A Ericsson2, CL Franklin2 are constantly expanding and require additional space for housing. This limitation has compelled some programs to seek housing 1Center for Comparative Medicine, Brigham and Women’s Hospital, options that maximize the use of their existing space. One company’s Boston, MA; 2University of Missouri, Columbia, MO; 3Boston optimization solution is to use “pie-shaped” cages placed on a University, Boston, MA rotating carousel rack. This creates a smaller footprint per 100 cages as compared to traditional shoebox caging systems. In this design, An animal care program is considering a change in mouse bedding the traditional exhaust blower system is replaced with a passive air substrate, being one of the few institutions using a pine-based exchange system. We predict that this will benefit breeding due to the bedding (alpha chip). Changes that effect the animal’s lack of noise and vibration typically associated with individually microenvironment may introduce new experimental variables to ventilated caging racks. Furthermore, 2 or more cages in a row can be ongoing research which is of concern to our research base. Prior to connected via an accessory piece of tubing to form a larger housing making decisions about bedding substrate change, we investigated unit. This allows for a greater number of animals per housing unit the effect of bedding substrate on mouse gut microbiome and while maintaining equivalent density. This format mimics extended operational impact for 5 commercial substrate types (alpha chip, beta family groups, which is consistent with naturalistic social conditions. chip, aspen chip, performance blend, and corn cob). Microbiome Here, we investigate the effect of this caging system, in either single analysis was conducted using 8-wk, female, CD-1 outbred mice from

645 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 the same vendor room housed 4 per IVC cage changed every 14 d. concluded that labs can store/use their excess bone wax for up to 37 Autoclaved full cage setups were washed and prepared consistently wk when either of the 2 methods are used. across all experimental groups with bedding substrate as the only difference. Animals were housed on the trial bedding type for 3 mo P151 Waste Not, Want Less: Being Efficient with Rodent Feed and then final fecal and cecum contents were collected for analysis. Without Compromising Care Microbiome DNA extraction, 16S rRNA amplicon library preparation and sequencing (Illumina MiSeq platform), and data analysis was B Stevens*, N Gooden performed by the University of Missouri Metagenomics Center and University of Missouri DNA and Informatics Research Cores. No Laboratory Animal Care Unit, University of Tennessee Health Science statistical differences between bedding substrates were realized in Center, Memphis, TN microbiome richness (estimated by Chao-1 index) of feces (n=8 mice/ group) or cecum (n=4 mice/group) contents based on one-way One of the biggest expenses for any rodent facility is feed. Our ANOVA. beta- (between samples) diversity modeled with principal department focused on how we could reduce costs without affecting coordinate analysis (PCoA) using several data normalization the level of quality care given to the animals. We focused on mice in techniques did not show significant shifts in the microbiome ventilated caging. Initially we collected, weighed, and evaluated the populations after exposure to different bedding substrate amounts of discarded feed from each of our rodent facilities from the (PERMANOVA). Based on this study we believe a change in bedding complete rodent cage change outs performed biweekly and noticed a substrate will not greatly impact mouse gastrointestinal microbiome significant amount was being discarded. To determine how much and downstream phenotypes influenced by the microbiome. feed was actually being consumed, we used groups of C57BL6 and BALB/C mice to measure the amount of feed every 24 h for a few P149 Prevalence of Syphacia obvelata in Mouse Colonies of 7 weeks in order to discover daily consumption in relation to the Conventional Animal Facilities in Mexico City number of mice in a cage. To determine the new amount of feed needed per cage we used our maximum number of adult mice AP Antunez*, C Cabello-Hernández, E Aguirre-Hernández, A allowed per cage and the frequency we normally change cages out (5 Carmona-Castro mice for 7 d). From these observations and measurements, it allowed us to decrease the initial amount of feed being fed to each mouse faculty of sciences, National Autonomous University of Mexico, cage. We also use a more accurate, standardized measuring container Mexico, Mexico so that the amount of food that was fed was relatively the same for each cage. Our technicians monitor the feed daily as part of their The presence of intestinal parasites is a persistent problem in colonies daily animal health observations and normally add feed only when of laboratory mice. The pinworms Syphacia obvelata and Aspiculuris weekly cage changes are performed. While still maintaining quality tetraptera are the helminths most commonly found in the intestine of animal care, our department is projected to save over $70,000 in feed laboratory mice. The objective of this work was to determine the expenses for the year. prevalence of Syphacia obvelata in 7 conventional animal facilities located in Mexico City, since it is common that animal facilities P152 Efficient, Effective, Reusable, and Customizable Social exchanged between different production centers, which facilitates the Housing for Chickens in the Laboratory spread of parasites from one place to another. At least 5 samples were obtained from each animal facility. An average of the number A Augustin1, MA Crabbs*1, MC Sour1, BS Carney1, DM Isaacson1, M of eggs in the samples of each animal facility was found, identifying Sauer2,1 at the end a prevalence of pinworms of 57%. These results show that despite having basic biosecurity measures, we still have to work on 1Laboratory Animal Resources, Iowa State University, Ames, programs that minimize the risk of entry of these parasites. IA; 2Office of the Vice President for Research, Iowa State University, Ames, IA P150 Save Your Wax! Effective Storage Methods for Maintaining the Sterility of Unused Bone Wax We needed caging specifically designed to socially house chickens in the laboratory. Previous caging consisted of components from other AD Pavan, J Stewart*, RA Malbrue, K Emmer, VK Bergdall caging types and were not always available. This caging was difficult to clean and assembly was time consuming and labor intensive. University Laboratory Animal Resources, The Ohio State University, Fiberglass reinforced plastic (FRP) wall panels are used to create Columbus, OH inexpensive walls that can be handled easily by 1 person. The walls are trimmed to optimal height with the remaining pieces used as wall Aseptic technique improves surgical outcomes by keeping microbial extensions, animal dividers, or platforms for feeding and watering. contamination of the surgical site to its lowest level. To maintain Stainless steel wing nuts, bolts, and washers are used to fasten walls. asepsis, surgical supplies are sterilized inhouse or purchased Holes are drilled in the walls along the short edge using a template to presterilized. Bone wax, most commonly used in in murine ensure interchangeability and facilitate the fastening of multiple intracranial surgeries for defect closure, is purchased presterilized in panels. Additionally, holes are drilled along the top edge of the walls a foil packet. Only a small portion of the 2.5g of bone wax is typically for securing the PVC frame and snow fence using zip ties and gear used during the procedure. Postsurgery, unused bone wax must be ties for access points. A frame constructed of PVC pipe is used to disposed of as it is no longer considered sterile. This results in support the snow fence lid, to hang heat lamps and enrichment monetary losses and waste for researchers. Traditionally to avoid devices, and to help maintain the general shape of the cage. The such losses, labs stored excess bone wax in the original foil packaging snow fence is secured to the walls using zip ties with reusable gear and placed it inside of a nonsterile reclosable plastic bag for later use. ties used at the access points. Plastic sheeting is used as the floor of However, this does not comply with our institutional survival the cage and is secured to the outside of the walls using duct tape. surgery policy. The purpose of this study was to reevaluate our The prototype cage proved functional, customizable, easy to policy by assessing the sterility of the traditional bone wax storage assemble and disassemble by 1 person, and withstood the rigors of and comparing it to a novel method of storage that we believe would use including cage washing. The cage easily stores flat when not in result in less microbial contamination: store excess bone wax in use, requiring little space. The plastic floor collects all waste within sterile micro centrifuge tubes. Bone wax sterility was evaluated at wk the cage and greatly reduces cleanup time. We created inexpensive, 3, 15, and 37 with ATP bioluminescence and RODAC plate testing. reusable, customizable, and replicable social housing for chickens in All RLU measurements and RODAC plate growths sampled were the laboratory. below common thresholds acceptable for sterilized surfaces. Positive controls were not utilized. Based on these findings, we have P153 Withdrawn

646 Abstracts of scientific papers 2019 AALAS National Meeting

P154 Water Dispensing Guns: Use, Sanitation, Validation, and growth and spawning would be the (E) or (E+G) diet. These diets Storage helped the fish reach breeding size quickly and also helped them produce more eggs. B Clancy*1, A Sparks2, D Harrison2, C Martin2, JM Hickman-Davis2 P156 Comparison of Absorption Capacity and Ammonia 1University of Illinois College of Veterinary Medicine, Urbana, Production in Ventilated Mouse Cages between Compressed Paper IL; 2University Laboratory Animal Resources, The Ohio State Bedding and Ground Corncob University, Columbus, OH B Pallas*, S Steinman, J Sanders, ZT Freeman The Guide for the Care and Use of Laboratory Animals recommends that there should be a mechanism in place to ensure that automatic Unit for Laboratory Medicine, University of Michigan, Ann Arbor, watering systems do not harbor bacteria or debris in watering MI devices. Water dispensing guns (WG) with quick disconnect assembly were used to manually fill water bottles and to dispense Increased concentrations of ammonia within laboratory mouse cages water for rodent medicine treatments. WG were stored in clean cage contribute to adverse health consequences. We previously wash, treatment rooms, and animal rooms. The aims of this project demonstrated that ventilated mouse cages bedded with compressed were to develop an identification system for WG, standardize the paper bedding (CPB) significantly reduce the intracage ammonia sanitation process, and determine best practices for storage and level, when compared to corn cob cages. We sought to understand sanitation interval. Unique numbered sanitation cards were placed how CPB alters ammonia levels in ventilated mouse cages. We on each WG for tracking. Command hooks were used to hold WG hypothesized that the CPB has a significantly higher absorption out of the way and off the floor. Total amount of water in disconnect capacity than corn cob and is able to modify ammonia production lines with WG was measured using a graduated cylinder and timed within the cage. We evaluated the total absorbent capacity of the two with a stop watch. For water cultures, 1mL of water from each WG beddings by immersion in water (20 ml, n=5 per bedding, per time was incubated in lactose broth for 48 h at 35 deg C and the presence point). CPB absorbed significantly higher levels of water than corn of turbidity/gas was considered positive. Positive cultures were cob at both 5 and 60 minutes of contact time (P = 5.82 e-8, P = 5.82 repeated once to correct for sampling error. Water was collected from e-8, respectively). We next measured the instantaneous absorption by WG for culture before and after flushing. The sanitation process was dripping a fixed volume of water (20 ml) through a column of standardized using flush stations in clean cage wash and bedding which allowed nonabsorbed fluid to pass through. CPB concentrated bleach. All WG were sanitized and tested after 2 weeks absorbed on average 87.0% of the water which was significantly and then monthly for 6 months. Initially only 2% of WG were more than corn cob at 40.9% (n=5 per bedding, P = 0.007). To positive. The longest amount of time to completely flush the water determine how ammonia production differed between bedding line was 17 sec. Flushing WG for 20 sec before use decreased positive types, ammonia levels for mouse cages controlled for age, sex, and culture results by 66%. We recommend that 1) WG be stored away number of mice (n=13 or 14 cages for CPB or corn cob, respectively) from surfaces and floors; 2) stored water lines should be air purged were measured. Cages with CPB had lower ammonia compared to to avoid standing water; 3) WG should be flushed for 20 sec prior to corn cob bedded cages at 14 d postcage change (P = 0.054). Next, use and extension hoses should not be attached; and 4) WG should mice and lids were removed from cages to standardize ammonia be tested quarterly and sanitized every 6 mo in the absence of a concentration across the bedding types, and lids were replaced. positive culture. Ammonia was then measured at 2, 4, and 17 h after cage change to assess the ammonia productive capacity of each bedding type P155 Finding the Best Feeding Strategy for Optimal Growth and without animals present. We observed that corn cob cages had Spawning in Zebrafish Danio( rerio) elevated ammonia over all time points (P = 0.020). Corn cob cages also had an increased rate of ammonia production over the first 2 B Unverzagt* hours (P = 0.054). We further found that this rate of ammonia production correlated with the ammonia levels measured prior to Friemann Life Science Center, University of Notre Dame, Notre cage change (RHO = 0.87, P = 4.05e-9). These data show that CPB has Dame, IN a higher absorptive capacity than corn cob, and suggests that this property functions to inhibit the process of ammonia production In their natural environment zebrafish consume a varied diet, many within ventilated mouse cages, and further supports its use as a of which are not accessible in a laboratory setting and as a result, may replacement for other traditional bedding types. not get all the nutrition required. This can be detrimental to their growth and spawning potential. To determine which commercial P157 Innovative Smart Cages for Improved Animal Welfare foods are best for growth and spawning, a feed study of 5 different food combinations was designed. The experimental feed groups CA Hall* included: the first group (G) was fed a diet made up of high- percentage soluble hydrolysed marine proteins, HUFAs, Comparative Medicine, Pfizer, Pearl River , NY phospholipids and algae; the second group’s diet (E+G) was composed of the first diet and an artemia- and rotifer-enriched brine To provide high-quality husbandry for the many cages of mice in our shrimp (instar 2); the third group’s diet (S+G) was composed of the facility, we introduced smart cages to help refine our practices. The first diet and brine shrimp (instar 1); the fourth group’s diet (E) was smart cage uses electromagnetic field technology to monitor the made up of artemia- and rotifer-enriched brine shrimp (instar 2) only; bedding condition, animal activity, and water levels of each cage. It and the control group (S) was fed brine shrimp (instar 1) only. Fifteen can also provide real-time census and monitor feed levels. The cages fish from each feed group were weighed and measured once a week require a learning phase, the period when data is collected, which is for 10 wk. The results indicated that fish fed E diet, S+G diet or E+G used to build an algorithm based on the facility’s standards. We diet, reached reproductive maturity length of 23 mm 3 wk sooner housed 280 mice in 80 smart cages using various numbers of mice than the G diet and 1 wk sooner than the S diet. The fish fed S+G diet per cage. The cages were checked daily for bedding conditions to or E+G diet also weighed 67% more than the G diet, 39% more than determine if the cage needed to be changed. If so, the cage was the S diet and 20% more than E diet. After the 10 wk, 6 females and 4 removed from the smart rack, changed, and put on into a males from each feed group were selected for spawning. The eggs conventional rack. Once the last cage on the smart rack is changed, were collected and counted for 10 spawns. We found that feeding E the learning phase is finished. The learning phase is usually diet or E+G diet, produced 140% more eggs than G diet, 75% more concluded within 6 wk , and the algorithm is then constructed. The eggs than the S diet and 39% more total eggs than S+G diet. In smart cages are now functional with the capability to notify conclusion, the results show that the best feeding strategy for optimal personnel when feed is low, if there is a sudden increase in water

647 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 flow, increase or decrease in animal activity, and when cages need to animal facilities for sanitization. At our institution, approximately be changed. Alarm notifications are done automatically by the smart 46 bottles of diluted quat are used throughout the vivarium to cage software via email. This technology allows for cages to be facilitate hard-surface sanitization. While reviewing the quality changed only when required, instead of as prescheduled cage assurance program, it was discovered that quat bottle refilling changes. Furthermore, the software linked to the rack also plans out was not on a formal schedule. This was not consistent with the cage changes and provides an easy way to collect census and quickly vivarium’s sanitation practices. The chemical manufacturer and identify animals. The animal activity data can be used in research for supplier recommend daily mixing of quat dilutions which is cost- studies that involve challenging models and provide additional data and time-prohibitive. Quality control ATP testing throughout the to compare groups within a study. This product enhances animal facility suggested quat solutions remained effective beyond 24 h. welfare by providing a secondary method to track animal activity. Therefore, a study to determine the concentration of diluted quat When animals are sick, activity is significantly decreased. The smart solution over time was undertaken to determine if bottles could cages alerts technicians providing early detection to health conditions be refilled less frequently than every day. Forty, identical, opaque and early response time to steady water flow. The smart cage trigger-spray bottles were filled with approximately 975ml of quat supports the refinement principle of the 3Rs. It also improves facility diluted to at least the recommended concentration of 660ppm using workflows and reduces resources form both a cost and labor a calibrated proportioner. Two additional bottles were filled to act perspective cages with 5 mice per box are changed on an average of as controls, 1 with water and 1 with concentrated quat. Bottles were 16 d compared to every 7 d. This results in cages being changed numbered and tested with quat test paper and with a titration kit to approximately 23 times a year opposed to 52 times year with a confirm the starting concentration. All diluted quat bottles initially conventional rack. Therefore, reducing labor hours used in animal tested between 700-850ppm while the water tested at 0ppm and room and cage wash, water used in cage wash, bedding and energy the concentrate at greater than 1000ppm. The bottles were stored used in cash wash by approximately 55%. in a temperature-controlled, secure area with a 12/12 light/dark cycle. They were randomized and tested for concentration with quat P158 Integration of Cage-based and Flexible Film Germfree/ test paper daily and by titration at least weekly. All diluted quat Gnotobiotic Isolator Systems to Establish an Adaptable and bottles remained above 660ppm for at least 30 d. The institution’s Efficient Small-scale Germfree Rodent Facility policy for quat bottle filling was subsequently altered to reflect the study results. Bottles are now emptied, refilled, and date-labelled C Umana*, A Discua, J Stathopoulos, T Caron monthly resulting in fewer personnel hours with a cost savings of $200 per month over weekly preparation and $2,000 per month over Comparative Medicine, Broad Institute, Cambridge, MA daily solution preparation. This bottle preparation schedule does not require additional chemical purchase for the department and is In response to growing research demands, many institutions are manageable for staff while maintaining optimal sanitization. pursuing the establishment of small-scale rodent germfree and gnotobiotic research operations within existing laboratories or P160 How Implementing a Green Committee in an Animal Care vivarium spaces. To ensure the ability to adapt as quickly as possible and Use Program Can Decrease Waste while Increasing Employee to the ever-changing needs of our rodent researchers, we have Engagement and Fulfillment established a new, small-scale germfree and gnotobiotic facility based around an integrated system that uses both flexible film isolators and D Flanagan*, A Heiser, E Moeller individually ventilated caging systems. These 2 housing systems present unique benefits and drawbacks when used in smaller Center for Comparative Medicine, MGH, Charlestown, MA operations. For instance, when exclusively using our flexible film isolators, we can house up to 1,200 mice in our 284-sq-ft housing Hospitals account for almost 6 million tons of waste in the U.S. room, with 25 to 60 mice sharing a primary atmosphere, while when annually. This waste is either burned or collected in landfills and exclusively using individually ventilated cages in the same space, we takes decades to break down. Research programs are no exception to have the potential to house 2,950 mice, but are limited to 5 mice this waste stream, but sustainable reduction is difficult to implement sharing a primary atmosphere. Furthermore, when made to our for various reasons including increased time, resources, education standards, flexible film isolators take 3 wk to prepare and validate, and difficulty finding sustainable products. In February 2019, we while individually ventilated cages take only 48 h. In our facility, established a continuous improvement team, called the Green these housing systems are prepared, taken down, or combined as Committee, to identify opportunities to reduce, reuse, and recycle. needed, allowing us to select the most appropriate setup based on They were empowered to find solutions to the current waste streams, experimental requirements (e.g., flexible film isolators for activities communicate with outside refineries about recycling, and educate such as large scale studies and breeding, and individually ventilated the rest of CCM on not only the processes of reducing, reusing, cages for small pilot studies and experiments requiring increased and recycling, but also the impact on the environment, animal, and dexterity) and timeframes. Developing a workflow for use between human health. The Green Committee educated the staff through these 2 housing systems has required the establishment of novel emails about the importance of participating in optional sustainable methods for maintaining germfree or gnotobiotic status during the practices. A survey was given later to determine participation, receipt and initial testing of new animals, the transfer of germfree/ employee engagement, and fulfillment. The committee implemented gnotobiotic animals between different housing systems, and during several initiatives including the use of a reusable single cup coffee the execution of experimental manipulations. Use of our methods brewing container in the break room, along with reusable dishes and and principles may offer established and developing small scale mugs and provided a drying rack to allow staff to wash their dishes. germfree rodent facilities ways to integrate new and existing housing After consulting with the Environmental Services Department, it was systems, expediently satisfy researcher needs, and streamline determined that cage cards and mouse food bags were recyclable workflow. material. Tracking of use showed that euthanasia cards averaged 439 per week while food bags averaged 104 per week. The Green P159 Verifying Disinfectant Concentration over Time for Cost and Committee was able to combine these and recycle an average of Time Savings 277kg of paper products instead of having them enter landfills each year. Ninety-five percent of the staff surveyed either felt fulfilled or CJ Staunton*, T Frese that they were making a difference when asked about how having a Green Committee at their facility made them feel. Comparative Medicine Facility, Loyola University, Maywood, IL

Quaternary ammonium compounds (quats) are inexpensive and safe disinfectants for most surfaces and are widely used in laboratory

648 Abstracts of scientific papers 2019 AALAS National Meeting

P161 Reducing the Replacement of Elizabethan Collars in P163 Benefits of Creating an Ergonomic Committee Laboratory Rabbits (Oryctolagus cuniculus) after Surgical Procedures D Hyde*

A Zall*, N Freeman, A Slate Research Animal Resources, University of Minnesota, Blaine, MN

Center for Comparative Medicine, MGH , Charlestown, MA Our department has a number of employees who have been impacted by ergonomic and repetitive-use injuries, and in this As part of the animal program, rabbits are fitted with Elizabethan industry, we are not alone. Repetitive motions are unavoidable in collars (e-collars) after surgery until their wounds are healed which is many of our daily duties, for example cage changing, twisting water typically 12-14 days. Husbandry staff noted that 68% (24/35) of bottle caps, and heavy lifting. Since we cannot evade all repetitive rabbits that damaged the e-collars did so within 9 d of placement. motions while performing daily duties, we have created an Complications that were noted as a result of this damage included Ergonomic Committee to come up with solutions to help reduce full removal with signs of self-mutilation of the surgical site or repetitive-use injuries, while also providing a more comfortable animals with their feet or mouth stuck in the placed e-Collar. This environment for our employees. The committee is made up of animal negatively impacted the animals’ wellbeing and resulted in extra husbandry staff, supervisors, health and safety employees, as well as work for husbandry and veterinary staff. In response to these issues, an occupational health doctor. The committee meets in small work a study was conducted to evaluate the use of destructible enrichment groups on a regular basis, along with larger meetings, that bring all as a potential deterrent to chewing the e-collar. Using an e-collar of the members together. Problems are identified and the groups damage scoring system, developed by the team, data was collected work together to brainstorm solutions to improve specific processes. on the number of e-collars destroyed by postsurgical animals with To date, 2 of our major accomplishments have been receiving saddle destructible enrichment provided immediately after surgery and chairs and improving the process of dumping water bottles. The compared to a group of postsurgical animals with our standard saddle chairs aide in comfort and ease for the husbandry staff to enrichment (stainless steel rattle, jingle ball, and dumbbell). Overall, perform health checks and while working in lowered biosafety there was an 11% (4/35) decrease in the number of e-collars that were cabinets. The bottle-dumping process has been changed so that the replaced for animals provided destructible enrichment. This decrease work is distributed to all staff instead of a select few. Because these resulted in a monetary savings of $13.85 as well as 30 min of staff changes are so recent, data collection and analysis is ongoing, but the time per collar replaced. staff have communicated how much the changes have impacted them in a positive way. The occupational health doctor is aware of P162 Can We Ever Achieve Reproducibility? Diversity of Rodent the changes and we all hope, and expect, to see a decline in work Husbandry between Collaborators injuries. Although the committee is still relatively new, it has made great strides in improving the work flow and creating innovative, DR Ferguson*, ML Granzow, FC Hankenson simple solutions to some of our biggest problems. The benefits from creating our Ergonomic Committee has exceeded our expectations, Campus Animal Resources, Michigan State University, Lansing, MI and we are excited to see what we will accomplish as we move forward. Research relies on repetition of experimental parameters for optimal reproducibility. Beyond study methods and reagents, it is critical to P164 Implementation of a Low-cost Electronic Rodent Health recognize the physiologic impact from housing and husbandry Records System practices for laboratory rodents; often these key details are left out of protocols and publications. Our institution has seen a dramatic rise in DJ Smith*, SM Kirchain, AF Hoggatt imports from collaborating institutions. We are on pace for 4 times more rodent importations in 2019 compared with 2018. We sought to Center for Comparative Medicine, Brigham & Women’s Hospital, assess diversity in husbandry environments between the original site Salem, MA and our own, assuming that our practices differed from those of collaborators. As is typical, our incoming laboratory rodents first go Literature on laboratory rodent medical record management systems into a quarantine area. They are provided with aspen chip bedding in is sparse. The computer spreadsheet is a popular tool for static filter top caging, ad libitum irradiated fenbendazole feed, coordinating information across a team, often developed inhouse to antiparasiticide on cotton balls, and given bottled reverse osmosis suit specific needs. While inexpensive and simple to create, (RO) water. Once rodents are health-screened and approved to enter spreadsheets lack robust sharing capabilities, data validation, and campus colonies, husbandry and housing change from those used in dynamic reporting tools. Many commercially available electronic quarantine (e.g. change of feed, enrichment, caging, and bedding). medical record systems have such capabilities but can be cost We designed an optional survey as part of our importation prohibitive and require external technical support to customize. To paperwork to gain feedback on diversity of husbandry practices at address the shortcomings of these options, our veterinary team originating locations. Since the survey launch, responses were customized a web-based data capture system (built from an academic received from 16 collaborating sites. Institutions indicated using hospital consortium platform) for rodent health cases. While the autoclaved ventilated (n=14) or static (n=2) caging. Bedding types platform was intended to capture clinical trial data, its simple included corn cob (n=6), aspen chip (n=4), paper-based (n=4), alpha customization and inexpensive cost made it an ideal option for our chip (n=1), or a combination of aspen/alpha chip (n=1). More than needs. Designed with a modular point-and-click interface and half (n=9) use autoclaved bedding. Water types included RO (n=9), supported with detailed online tutorials, it is simple to use even for tap (n=2), chlorinated (n=2), acidified (n=1), filtered (n=1), or a individuals without programming knowledge. The user enters a case combination of treated water types (n=1). Enrichment and feed types number which prompts a custom-made questionnaire (answers varied greatly between institutions, however, cotton nestlets (n=13) validated and mostly multiple choice to reduce potential errors) for and a standard rodent diet (n=6) were the most commonly capturing animal signalment, clinical conditions, treatments, and mentioned. In summary, rodents experience a minimum of 3 cage information for future reporting. A notification feature for divergent husbandry environments (origin, quarantine, destination) generating standardized emails, autofilled with information from the in the context of moving between collaborators. To better promote corresponding record better coordinates communication between transparency for publications and clarify experimental variability, we members of the veterinary team and research groups. After a period advocate that institutions continue to discuss the diversity of of adjustment, this case management system has proven more husbandry and housing environments within the biomedical efficient for case entry (no need to retype redundant information or communities that use shared rodent models. write out specific conditions and observations when there are comprehensive multiple-choice lists) and transformed the team’s

649 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 ability to review health trends. Access to user-friendly histograms Food grinding, or chewing up of food without ingestion, is a recent and pie-chart displays of collected data (summarized by location, phenomenon plaguing the laboratory animal industry. Recently we protocol, users and conditions over selected periods of time) have observed a significant number of sentinel mouse cages (38 out of 71 replaced previously overwhelming amounts of paper exams and total cages) grinding up their food in 1 of 2 caging systems. The difficult-to-analyze spreadsheets. The ability to consistently review prevalence of grinders in our facility has increased the strain on our trends at weekly clinical meetings allows historical health data to resources and labor due to frequent cage changes and additional food function as a management tool, affording opportunities to improve provided. The purpose of this experiment was to see if various operations or better address research groups. enrichment strategies can mitigate the grinding behavior. Sixteen cages (8 of which were known grinders) of our CD-1 sentinels were

P165 Comparison of Ammonia and CO2 Levels in Static Mouse placed in 1 of the 4 treatment groups. The first group received only Cages Bedded with Corncob and Commercial Paper-based Material standard enrichment, a cotton square, and 4 oz puck of crinkle paper. Groups 2, 3, and 4 received standard enrichment plus a wooden D Keys*, B Pallas, J Szcodronski, J Sanders, S Bruckner, ZT Freeman block, 2 tbls of sunflower seeds, or both a block and seeds respectively. Once a week for 13 wk the food level in the hopper was ULAM, University of Michigan, Dexter, MI scored and the experimental enrichment was added. There was no significant difference in the grinding behavior with respect to the The Guide suggests that bedding is an important component of the enrichment group. However, there was a significant difference rodent cage environment that may alter intracage parameters such as between the nongrinding group and grinding group that received

ammonia and CO2. We previously found that paper-based bedding sunflower seeds. Sunflower seeds may have potential to prevent decreased intracage ammonia levels compared to corn cob bedding grinding, however we have not implemented any changes to our for mice housed in individually ventilated mouse cages (IVC). Given enrichment program since the enrichment tested did not reduce the that static cages tend to have higher ammonia levels compared to grinding observed. But we continue to look for a solution for this IVCs, we then hypothesized that the use of paper-based bedding in behavior problem. static cages would decrease intracage ammonia levels. We randomized cages containing adult mice into 1 of 4 bedding groups P168 Manipulation of Room Humidity for Optimal Embryo Donor (1x, 1.5x, 2x paper-based bedding and 1x corn cob bedding, n=8 Yield cages/group, males = 5 cages/group, females = 3 cages/group). We *1 2 evaluated intracage ammonia and CO2 levels on days 3, 4, 5, and 6 DK DeLoach , J Gonzalez postcage change, and prior to cage change on day 7, repeated over a 3-wk interval. We found no effect on intracage ammonia or 1Comparative Medicine, University of South Florida, Tampa, 2 CO2 levels with any of the paper-based bedding amounts compared FL; Gene Targeting Core, Moffitt Cancer Center, Tampa, FL to corn cob bedding, when controlled for days post change, mouse sex, and repeated sampling for each cage (ammonia P = 0.54, 0.47, The Guide for the Care and Use of Laboratory Animals recommends a

0.91, CO2 P = 0.42, 0.67, 0.99). Intracage ammonia levels significantly wide range of acceptable humidity levels for murine vivariums from increased with time post initial cage change while CO2 levels did not 30-70%. Our embryos are collected to use for various manipulations, differ (P = 2e-16, 0.22). Mouse sex had no effect on intracage such as DNA transgenesis, embryonic stem cell targeting, and ammonia or CO2 levels (P = 0.57, 0.11). Taken together, this data CRISPR/Cas9. It was noticed that during a certain time of year when shows that paper based bedding in static cages does not alter the humidity increased in the mouse housing room to as high as 85% intracage ammonia levels in the same manner as individually the embryo yield from donor females would conversely decrease ventilated mouse cages. from 70% to 0%. The objective of this study was to determine if manipulating the humidity would have an effect on the embryo yield P166 Lifetime Feeding of Alfalfa Compared with Timothy Hay for from donor females. To rule out other variables as the culprit for this New Zealand White Rabbits decrease we checked and tested out a few other theories. We ordered the same strain and age of mice from a different preapproved E Paster2, DL Hickman*1 vendor, we used mice of different ages (9-11-wk-old) from the same 2 vendors, we relocated the donor females and stud males to another 1Laboratory Animal Resource Center, Indiana University, room in the same hallway and gave them a 1-wk acclimation period Indianapolis, Indiana; 2Edwards Lifesciences, Irvine, CA prior to use, we changed our injection times, and changed the injection amounts but left the concentration the same. All other In exotic animal medicine, owners are advised not to feed pet rabbits theories led to the same conclusion so we decided to further alfalfa-based diets because of concerns that the high calcium levels investigate the humidity that fluctuated during seasonal changes. It present in alfalfa hay may contribute to renal compromise later in has been previously documented that humidity and different seasons life. To date, this recommendation is based on empirical observations, do have an effect on the donor embryo yield. Since this is a variable not controlled studies. In this study, we obtained 9 female and 8 male that we noticed at our facility we decided to attempt to manipulate New Zealand White rabbits. Half of these rabbits were maintained on the humidity level to see if we could compensate for any seasonal an alfalfa hay-based diet for their entire lifespan (range 5-8 y), while change to determine our optimal humidity level. We used a the other half were maintained on a timothy hay-based diet. The dehumidifier in an attempt to bring the humidity back within the rabbits were assessed at least semiannually with a physical exam that range where the embryo yield was hypothesized to be the highest included a blood and urine sample collection. No renal concerns (maximum 70% humidity). The dehumidifier remained in the room were noted in any of the 17 rabbits over their lifespan. The decision for one month and temperature and humidity levels were recorded. to euthanize was made due to complications associated with Our preliminary finding report that highest embryo yield is within a cardiovascular disease, Pastuerella, or neoplasia. This study suggests humidity level of 35%-65%. that the chronic feeding of alfalfa based diet can be safely accomplished in New Zealand White rabbits without increasing the P169 Technical Services from Animal Care Technicians Contribute chance of development of renal complications. to Employee Retention and Program Support in Academic Programs P167 Grinding on the Last Nerve: Attempting to Curb Food Grinding in Mice E McCann*, E Miedel, R Engelman

D Margolies*, T Garcia, C Brown, G Simonek Comparative Medicine, University of South Florida, Tampa, FL

Office of Animal Care, Seattle Children’s Research Institute, Seattle, To contribute to employee retention and establish both career and WA 650 Abstracts of scientific papers 2019 AALAS National Meeting

skill development, we created a staffing structure that combines the P171 A Cooperative Approach to Veterinary Care and Husbandry roles of both animal husbandry and research technical service for Wild-caught Birds providers. Staff is responsible for husbandry and housekeeping of animal rooms. In addition they implement technical services that are E Wegner*, C Winnicker, R Ober typically done by researchers in academic settings. This approach is a win/win for academic institutions: the technicians learn valuable ICM, Columbia University, New York, NY technical skills which results in job engagement and retention, and PIs can focus on grants and less on study and personnel lab Housing of wild-caught birds requires a modified approach to management. The department also gets reimbursed for the technical husbandry and veterinary management. Our black-capped services provided (at $30/hr), which contributes directly to chickadees (Poecile atricapillus) and tufted titmice (Baeolophus bicolor) subsidization of the technician’s salary. The technical service are wild-caught, therefore susceptible to stress until acclimated to life revenues annually reimburse 25-30% of our own animal care in captivity. Initially, approximately 10% of our colony was personnel expenses, contributing to program operating efficiency succumbing to opportunistic bacterial infections. Bird husbandry is with an average of 528 hours/month of billable technical assistance. labor intensive. Feed and water is changed daily, multiple feeds are Due to this revenue stream our institution can employ more provided (live, pelleted, and seed) and cage bottoms are changed 3 technicians per rodent box than the standard 800-1,000 boxes/ times weekly. Communication between veterinary, husbandry, and technician. In addition, allowing technicians to vary their daily duties lab staff was vital to implementing a coordinated care and between husbandry and technical work helps decrease repetitive management plan to lower stress induced by human presence. The work injuries. Implementing this system for the labs has ensured light cycle was gradually changed to 2 h prior to the start of employee retention, with an average retention of 9.5 y among 28 husbandry to maximize uninterrupted time for the birds to feed prior fulltime staff members, as well as increased employee development, to the first human disturbance. Husbandry procedures were with 100% of animal care staff AALAS certified, most (50%) at the performed at a consistent time to allow birds to acclimate. highest LATG technologist level, with 6 staff members also certified Husbandry materials and medications are prepared beforehand and by the Academy of Surgical Research and/or State of Florida as brought in together to allow single entry/exit per room. This Certified Veterinary Technicians. Finally, with animal care staff doing decreased the amount of time that staff were present in the room by many of the procedures involving animals for researchers, uniform half and improved efficiency. Additionally, daily rounds were standards of implementation are ensured, protocol compliance is coordinated between veterinary and lab members to check the colony assured, refinements are developed, and a mutually beneficial simultaneously, minimizing disturbance as much as possible. This collaboration between our care staff and the researchers has been coordination was achieved by planned meetings between the lab, formed. veterinary, and husbandry staff, and on a daily basis using text-based apps. The birds responded well to these adjustments; they now have P170 Are 2 Halves Better than One? more time to eat and groom undisturbed, and the incidence of opportunistic infections in the colony decreased to less than 3%. Once E Green*1, J Naden1, K DeBethizy2, S Garmon2, j nelson2 the birds enter the research paradigm, in which their natural food caching behavior is quantified, the lab acclimates the birds to single- 1Veterinary Science, Research and Support, Envigo, Indianapolis, housing; as a result, human interaction time remains contained to a IN; 2Production, Envigo, Indianapolis, IN short period daily. Communication and coordination between veterinary, husbandry, and lab staff is essential to managing The use of enrichment is important for laboratory rodent models to nontraditional species. reduce stress, abnormal/aggressive behaviors, and provide temperature regulation. When used properly, enrichment can lead to P172 Reducing Noncompliance in Clinical Assessments of Rodent a reduction in variability in research outcomes, reducing the number Ocular Disease: Combination of a New Method of Preparing of animals needed for a study, as well as improve animal welfare by Fluorescein Stain and Training promoting species-specific behaviors. We evaluated the efficacy of a half-sized twisted paper enrichment in 3 strains/stocks of mice: E Adams*, S Lewis C57BL/6NHsd, Hsd:ICR (CD-1®), and Hsd:Athymic Nude-Foxn1nu. The proposed benefit of the smaller half sized-enrichiment is that it is UT Southwestern Medical Center, Dallas, TX able to fit in most cages regardless of their size. Both inventory and breeding cages were evaluated for 12 wk to ensure that substitution Eye issues are commonly observed in laboratory rodent colonies. As of 2 half sized-enrichiment pieces in place of 1 full-sized twisted these patients typically present with general ocular symptoms, the paper enrichiment did not cause any distress to the animals, standard of care when assessing these abnormalities should include negatively impact their fecundity, or inhibit their innate nesting using fluorescein stain to ascertain if the cornea is ulcerated. This is behaviors. Nests were scored weekly according to the following critical to determining if topical medicine is necessary, or if the defect system: 0-5; 0-undisturbed, 1-disturbed, 2-flat nest, 3-cup, is deep and/or painful enough to warrant enucleation. After initial 4-incomplete dome, or 5-complete dome. There was 100% use of the training to adopt fluorescein staining as standard practice amongst assigned enrichment product in all mouse groups, however, the our veterinary technicians, follow-up monitoring of cases by the extent of usage varied by strain. While Hsd:Athymic Nude- veterinarians discovered lack of staining in every ocular case. The Foxn1nu had scores ranging from 1 to 5 for both the inventory and main reasons for noncompliance were that the full strips would not breeder cages with the majority of nests being cup or dome shaped, fit into 1ml syringes if the technician had run out of 3ml syringes, the the C57BL/6NHsd and Hsd:ICR (CD-1®) inventory cages had flat or number of individual strips carried would not always be enough for no nests with scores ranging from 1 to 2.75. No statistical significance the full day of cases, and if they had run out of saline on hand, the was observed in nest scores between the cages containing the full- step would be skipped. The use of them was considered cumbersome sized or half-sized enrichment within the stocks/strains. The number and time-consuming. To address this, a system was created to and size of pups per litter was recorded in the breeding cages and prepare the fluorescein stain in advance, so that administration was weekly body weights were assessed in the inventory cages. There as easy as possible and enough stain for the day would be available was no difference in litter size or body weights between groups. without returning to resupply on strips or syringes. Each strip (stain There were no aberrant findings, morbidity, or mortality related to portion only) is cut into 5 equal pieces and preloaded into 1ml or 3 the use of the full-sized or half-sized enrichment. Studies such as this ml syringes, fitted with syringe cap. The syringes are prepared at the will enable us to determine the best practices for nesting enrichment cleanest room level available for each facility, including under a hood in laboratory rodent models to better care for our animals and in the barrier facilities, with scissors cleaned aseptically. The promote the 3Rs. technician can simply take 1 syringe each morning and fill with sterile saline, then use the solution for all cases that day. Training was

651 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 implemented on how to administer the fluorescein solution without and improve the quality of decontamination. To mimic the touching the syringe to the body or eye of the patient, and to cap it challenging environment of aquatic facilities, all tanks were tested in when not in use, so that it could be kept free of contamination and the presence of biofilm without the benefit of presoaking. We therefore used on multiple patients. Retraining was also done to compared wash only, fog only, and wash + fog methods. Three forms reemphasize how essential this step was to diagnosing ocular of validation were measured: ATP swabs, aerobic swab testing, and disease. The method of preparing the fluorescein strips has simplified biological indicators of Geobacillus stearothermophilus. While wash day-to-day assessment of ocular issues and increased veterinary only demonstrated a reduction in colony forming units (CFUs) as technician compliance with our standard diagnosis guidelines across well as ATP, the most significant reductions were noted with fog only our program, with the additional bonus of reducing the waste of a and wash + fog methods, with wash + fog succeeding in eliminating full fluorescein strip per rodent. virtually 100% of CFUs. The fog only process outperformed wash only and significantly penetrated into the biofilm. In conclusion, the P173 Technician-led Team to Improve Ergonomic Awareness and 7% hydrogen peroxide pulse mist demonstrated a compelling Reduce Repetitive-strain Injuries improvement to the processes of decontamination in aquatic facilities. F Gallardo-Chang*, K Veliz, T Chen P175 Food Contamination in Rodent Metabolic Caging Laboratory Animal Resources, Genentech, South San Francisco, CA H Roeder*, P Lachcik Working with animals in a vivarium or laboratory setting often involves repetitive tasks that pose ergonomic risks. Ergonomic injury PKPD, BioAnalytical Systems Inc., West Lafayette, IN can lead to pain and discomfort, and in extreme cases, permanent disability. These outcomes not only affect staff health and wellbeing, Laboratory rats have long been used as a proxy to study human but can also cause staffing issues due to physical limitations and biological processes and serve a particularly important function in absence due to injury. Therefore, it is important to take a proactive preclinical drug development and biomarker discovery. approach to reduce ergonomic risks as much as possible.We formed a Biotransformations, uptake, and production of metabolites are all team of ergo advocates that works closely with our corporate biochemical pathways of metabolism that can be studied by ergonomists to devise best practices, brainstorm ideas, test new employing metabolic caging that allows for the separate collection of products to evaluate them for ergonomic potential, and serve as both feces and urine. A common problem with metabolic caging is resources to the rest of our group. Our best practices are cataloged in contamination with food particles that drop through the mesh cage a website sorted by the general body part that they will mostly flooring. Although uncommon, it is possible for food proteins to bind benefit. Some of the tips and best practices we devised for wrists, the to protein biomarkers which can decrease the amount of biomarkers area most commonly injured, were keeping a natural wrist position found during routine analysis and thus misrepresent results. Here, 2 while holding tools like an ear notcher or tail clipping scissors, methods were employed to decrease the amount of contamination. alternating hands for scruffing animals and grasping tools, and First, a wire-barred insert was developed to slide into the cage’s food switching the computer mouse to the side of the nondominant hand. hopper. Second, a diet gel was compared to a traditional pelleted Some of the general best practices include dividing out specific tasks diet. The urine output was then ranked on a scale from 1-5 each for throughout the week and taking micro-breaks. In addition to best volume, clarity, debris, and color with 1 being the best. The wire- practices, we also created our own stretching posters focused on barred insert deterred most subjects from removing all the food from body parts most commonly fatigued and injured. We have their food hoppers at once. When using a pelleted diet, the barred encouraged these stretches by distributing the posters throughout insert (n=14) marginally increased the urine quality (4.1 to 3.5) when our facility and leading group stretches twice a month. compared to an open food hopper. Exchanging pelleted diet (n=15) for diet gel (n=10) in an open food hopper also improved the average P174 Effective Penetration of Dry Biofilm with Pulse Hydrogen urine quality (4.1 to 2.1). Finally, by using the wire-barred insert and Peroxide the diet gel simultaneously (n=10), the average urine quality was greatly improved (4.1 to 1.6). In conclusion, using both of these C Greene3, FM Grinstead*1, L Pandolfo2 methods decreases food contamination and generates better quality samples. 1CURIS System, Oviedo, FL; 2NIH, Bethesda, MD; 3NSF International, Ann Arbor, MI P176 Withdrawn

In most aquatic facilities, a heavy gross contaminant of biological P177 Hide and Go Squeak: A Different Approach to Technician matter, known as aquatic biofilm, adheres to tank surfaces. Biofilm is Training and Performance Assessment necessary to the health of aquatic environments; however, pathogens inhabiting biofilm may compromise the health of fish, threaten HN Splawn*, M Pachucki, J Criley scientific data, and pose a danger of cross-contamination. For example, Mycobacterium marinum is a known pathogen that is Comparative Medicine, Oklahoma Medical Research Foundation, commonly present in aquatic facilities, causing painful granulomas Oklahoma City, OK in extremities and resulting in long and painful healing time. Due to the continual accumulation of biofilm on tanks, there exists an Routine husbandry of laboratory rodents can be repetitive and drifts extensive and laborious cleaning regimen. Tanks are emptied of in performance can occur over time. There are numerous approaches water and allowed to dry for up to 2 mo, becoming considerably to monitoring training and performance—blending of these methods more difficult to clean and disinfect. Disinfection is essential since can provide robust monitoring of the quality of animal husbandry certain pathogens maintain the ability to reanimate if suspended in and care. Some of the traditional methods, such as supervisor dry biofilm. Currently, there are no known disinfectants that can observation or auditing against SOPs test knowledge of processes can substantially penetrate dry biofilm. In fact, various disinfecting be stressful and also do not monitor the performance of employees products such as chlorine dioxide, sodium hypochlorite, and not being directly observed, which is when drift can be most notable. peracetic acid have had challenges affecting even the easier-to-treat We sought to implement a system that would increase active wet biofilm. To determine if hydrogen peroxide fogging could engagement in all of the processes involved in animal care in a fun penetrate dry biofilm while improving efficacy over currently way without increasing the stress level of employees. We purchased implemented methods, we evaluated incorporating a pulse mist 7% small toy mice which were sterilized and labelled. The supervisors hydrogen peroxide system in 3 different treatment processes. and management developed a list of places to hide the mice targeting Successful outcomes would ease the burden of the cleaning process areas of concern for cleaning or observation. A table was created to

652 Abstracts of scientific papers 2019 AALAS National Meeting

log when and where mice were hidden and when they were found disinfectants, paper towels, food, cage cards, and logs. The choice and by whom. The toy mice were hidden in locations that technicians and placement of tools (e.g., card holders, paper towel holders, hooks are responsible for checking daily (such as empty mouse cages on the for clipboards) and positioning of other supplies (e.g., clean cages) rack and the pest traps) or cleaning and maintaining weekly (such as were determined based on technician feedback and efforts to the inside of the cage change hoods and on top of the racks). Mice minimize ergonomic concerns. Challenges encountered when were also hidden in general areas not assigned to a particular implementing use of the customized change stations included technician (such as storage areas and euthanasia stations). A point interference of researcher access to cage racks and select tools not system was developed which factored in how quickly the mice were functioning as initially envisioned. Multiple strategies were used to found after hiding, if the technician found all of their mice, and if the resolve these challenges including establishing a brief training technician found any other bonus mice in general areas. Technicians program for animal technicians unfamiliar with the process, creating could cash in their points for an assortment of prizes. The program custom hooks to hold supplies, and increasing communications with was well received and increased employee engagement. We believe research staff. Animal technicians recorded the time it took to that combined with traditional assessment methods the Hide and Go complete routine cage changes using both old and new processes Squeak program enhanced our employee performance by reinforcing from initial setup to end of cleanup. Use of the customized change training. station decreased unnecessary travel time and movement, with all movement now taking place in a ~3-sq-ft space. As a result of our use P178 Performance Qualification Testing of Rodent Drinking Water of the customized change stations and modified processes, we Systems anticipate increased efficiency of the cage change process and, based on observations, a decreased risk of ergonomic injury. JR Fahey*2, JM Vollmer3, R Malcolm1 P180 Efficient and Effective Approach to Gnotobiotic Isolator 1Comparative Medicine and Quality/ Jax Mice and Clinical Research Sterilization Using Chlorine Dioxide Gas Services, The Jackson Laboratory, Bar Harbor, ME; 2Comparative Medicine and Quality, The Jackson Laboratory, Bar Harbor, ME; 3JAX JL Ludwig* Mice and Clinical Research Services, The Jackson Laboratory, Bar Harbor, ME Division of Animal Resources, University of Illinois, Urbana, IL Assuring the sanitary status of water provided to mice is of great Ensuring sterility of gnotobiotic isolators, prior to the beginning of importance in preventing transmission of infectious agents and any microbiome research, is critical. Achieving overall sterility using subsequent disease outbreaks. Therefore, the design, care, and liquid chemicals can be a time consuming and difficult task. quality assessment of water treatment systems is vital to animal Challenges include maneuvering in restrictive gloves and ensuring biosecurity and play an essential role in maintaining high standards that every inch of the isolator is covered with the sterilant. We of animal welfare. The water provided to mice at our institution is simplify this process with the use of chlorine dioxide gas with a treated through an animal water purification system designed to portable gas generator. Chlorine dioxide gas is a true gas, and when remove microorganisms, metals, and suspended materials from introduced via a generator, easily reaches every surface within the water. The first step in assessing the performance of a water system is isolator. This method proves to be both efficient and effective. The to evaluate water quality over time. We undertook a performance isolators are constructed of polyurethane material and are completely qualification program of the water treatment system in a newly sealed when the port caps are in place. They are perfectly suited as completed mouse facility. The purification process was validated by individual chambers to accommodate the chlorine dioxide gas weekly sampling of water for one month during which samples were without exposure to personnel. Their individual filtration systems tested for pH, total organic count (TOC), heavy metals, specific and blowers allow for appropriate aeration using passive scavenging. conductivity, and microbiological quality. Water samples were A port cap is modified to accommodate our gas generator collected at several points throughout the system to monitor the connections. The isolator is prepared with autoclaved filters for both function of each component not just the final outlet. Post validation, the input and exhaust connections. Each filter is sealed with mylar the water system has been routinely sanitized and microbiologic film to maintain sterility until the isolator is ready for use. Biological quality assessed on a weekly basis. During operational testing over spore strips specific to chlorine dioxide are placed in the isolator the past 10 mo, this newly designed water treatment system has along with a small battery-operated fan to assist with circulation of consistently provided high-quality drinking water free of microbial the gas. Humidity is introduced via portable humidifier and serves to contaminants and well within acceptable limits for chemical soften spore walls to allow the gas to penetrate and destroy the spore. contaminants of surface water sources as shown in the data reported The isolator is then sealed with the pre-made cap and the gas here. Results from the weekly microbial testing during the first year generator is connected. The generator is programmed with the will be analyzed to determine if we can lengthen the interval chamber and appropriate cycle parameters. Each isolator is exposed between sanitizations. to chlorine dioxide gas to 720 ppm. Once the cycle is complete the isolator is connected to a modified carbon scavenger, the mylar on P179 Customized Rodent Change Stations: Benefiting Animal the filters is disrupted and the isolator can cycle, thus aerating the Technicians and Supporting Process Improvement gas by passive scavenging. Biological spore strips are then incubated, and once confirmed negative, further microbiological monitoring is J Kenfield*, T Barcelli, G Melanie, LA Colby performed to confirm sterility. Standard 4’ x 2’x 2’ and 3’ x 2’ x 2’ isolators can be sterilized using chlorine dioxide gas via portable gas Comparative Medicine, University of Washington, Seattle, WA generator in less than 4 h.

Animal technicians play a vital role in supporting the health and P181 Environmental Temperature Exposures of Laboratory Mice welfare of laboratory animals. Their jobs are often physically during Shipment strenuous and they are under constant pressure to perform tasks with maximum efficiency. Our institution’s routine rodent cage E Himrod*, E Wall, J Villano, E Syversen change process requires animal technicians to walk cages between a cage rack and a stationary laminar flow change station, a time- Unit for Laboratory Animal Medicine, University of Michigan, Ann consuming process that requires technicians to travel large Arbor , MI cumulative distances (average of ½ mile per side of rack). To increase efficiency and decrease technician effort, we recently instituted a Laboratory animals are routinely acquired from vendors and other process in which technicians move mobile change stations to each research institutions. Their transportation from 1 facility to another is rack. The change stations’ exteriors were customized so technicians an animal welfare consideration because of its stress induction. While have easy access to necessary supplies including gloves, sleeves, there are rules and regulations on environmental conditions for

653 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 transportation of common large laboratory animal species, there is a P183 Space Allocation and African Green Monkey Social Group lack of equivalent information for laboratory rodents. Here, we Introductions investigated environmental temperatures mice were exposed to during transportation from our facility in Michigan to 7 institutions JJ Breaux*, S Breaux, R Clark in the northeast, midwest, south, and southwestern regions of the United States in April and May 2019. Data loggers were placed in University of Louisiana, New Iberia Research Center, New Iberia, LA shipping crates of animals to record temperature at 1-minute intervals during the entire transportation process facilitated by the At our facility African Green monkeys (AGM) have historically been same local ground agent in the point of origin and airline. Results housed in breeding pairs or trios. Renovations of social housing were categorized based on the steps of the process: 1) origin, ground space recently afforded our facility the opportunity to transition agent; 2) origin, airline warehouse; 3) origin, cargo hold; 4) in-flight; to more naturalistic multi-male/multi-female groups of AGMs. 5) destination, cargo hold; 6) destination, airline warehouse; and 7) Introductions of 13 groups of unfamiliar AGMs in 2 types of indoor/ destination, ground agent. Data collected for all except step 4 were outdoor enclosures, small box cages (SBC; 195 ft2) or large box cages also compared to local (origin or destination) temperature data (LBC; 696 ft2), were conducted between December 2015 and January collected by weather stations. Step 4 data were analyzed to verify 2018. Multi-male groups were comprised of juvenile and adult male consistency in-flight cargo temperatures across shipments. Results pairs or trios or groups of 4 young adult males (~5-y-old) that were revealed that mice were exposed to an average fluctuation of 7.5°C, previously housed in bachelor groups. Groups of 6-8 unfamiliar with temperatures ranging from 7.4 ± 1.5 (step 5) to 26.6 ± 0.6°C (step females and 1-2 males were introduced in SBCs; groups of 11-22 7). Temperatures while on the ground varied widely and in-flight unfamiliar females and 2-4 males were introduced in LBCs. The temperatures ranged from 10.9 ± 4.0 to 22.5 ± 0.1. The most retention of individuals in newly introduced social groups were significant changes were seen during step 4, when temperatures compared between 2 sizes of indoor/outdoor housing. Average decreased 10-20°C for flights lasting at least 100 min, and then density was 22.9 ft2/animal in SBCs and 35.9 ft2/animal in LBCs. increased during steps 5 and 6. Temperatures in 4 shipments were In SBCs, 57.1% of animals introduced remain in the groups 6-13 mo not significantly different from local temperatures. In conclusion, postintroduction and 71.5% animals introduced in LBCs remain in laboratory mice could be exposed to a wide range of environmental the groups 1-5 mo postintroduction. This preliminary analysis may temperatures during transit. While the mice may be healthy upon suggest that unfamiliar AGMs can be introduced more successfully receipt, as in our study, it is important to monitor weather forecasts at when additional enclosure space is available, making the larger box both the origin and destination locations when scheduling shipments cages a better choice for new group formations. and to avoid extreme temperatures to minimize transportation- P184 Rust and Dust Be Gone: A Refined Protocol for Effective IVC induced stress. Plenum Sanitation

P182 Evaluation of Dental Rolls as Environmental Enrichment for A Sparks*, K Nolan, C Martin, D Harrison, D Coble, C Ayres, JM Mice Hickman-Davis

JC Rodgers*1, RJ Ricart Arbona2, C Lieggi2, N Lipman2 Ohio State University, Columbus, OH

1Center of Comparative Medicine and Pathology, Weill Cornell The Guide for the Care and Use of Laboratory Animals indicates Medicine, New York, NY; 2Center of Comparative Medicine and that visual inspection may be an important component when Pathology, Memorial Sloan Kettering & Weill Cornell Medicine, New validating cleaning and sanitation practices. Evaluation of sanitation York, NY practices on rodent ventilated (IVC) rack systems identified excess accumulation of dust and rust in 59% of vertical rack plenums. Nesting material is an important component of environmental Recommendations from the manufacturer included removal of rust enrichment for mice. In a large animal care program, costs and using stainless steel wire and application of a commercially available logistics are a major consideration. Currently, our technicians rust disolver followed by a water rinse. The manufacturer provided manually tear perforated sheets of compressed cotton squares and a 2-inch round head brush on a 6-foot flexible extension rod for place 2, 1 in x 1 in squares into each shoebox cage after it’s filled with removal of dust and encrusted debris. Detergents and chemicals bedding on a tunnel washer line. We implemented a pilot study to were not an option for plenum cleaning as these can corrode metal evaluate an alternative nesting material that would allow the mice to and contribute to rust development. The manufacturer brush offered build the same or better-quality nests while enhancing ease of minimal surface contact within the plenum and the flexible handle delivery. Three-point sized cotton dental rolls were evaluated as an caused the brush to bow away from the contact surface resulting alternative. The weight and cost of a dental roll is comparable to 2 in racks that continued to fail visual inspection. A large conical cotton squares and the distribution process is easier because no tube brush affixed to a long PVC pipe was created inhouse as part tearing is needed prior to placement into clean cages. A 2-wk trial of a brush and soak procedure to remove debris. The new brush was conducted to compare the nest quality with 1 dental roll (n=70 was large enough to ensure 360° contact with the plenum and long cages) versus 2 cotton squares (n=70 cages). Nest quality was enough to reach the end in a single pass. The PVC handle was determined daily using a nest scoring system (0–5; poor–best) over 7 sturdy enough to withstand the force of passage through the plenum d. Nests built using dental rolls (average score of 2.3) were of similar and featured a drainable endcap in case the tube filled with water. quality to those built with 2 cotton squares (average score of 2.4). The Plenums were flushed with high pressure water prior to brushing trial was then expanded to a room with 800 cages, all receiving a and the brush was dipped into a mop bucket of clean water prior dental roll weekly for 30 d. Feedback from cage wash technicians was to each passage. After brushing, racks were processed in a standard positive, indicating that the distribution of dental rolls was easier and rack washer and tilted to drain excess water. Development of the faster than cotton squares. Animal care and veterinary technicians brush and wash process for plenums allowed for an improved noted that mice take longer to make a nest with dental rolls (48 h) acceptable standard for IVC racks after cage wash. Rack plenums than with cotton squares (24 h). No other notable differences were must be 100% visually free from debris and rust in addition to observed so the trial was expanded to a 39-room facility (~10,000 passing the standard testing utilizing ATP luminometer and sterile cages). Within the first week of implementation, 9 adult mice were swabs for bacterial culture. The most important components for found with leg strictures associated with tangled dental roll fibers. effective removal of debris from IVC rack plenums was ample water Due to these adverse events, the trial was discontinued. We continue pre-soak and a large, sturdy brush to contact all surfaces prior to to evaluate nesting materials to determine the best option(s) for our washing in a mechanical rack washer. program to improve nest quality.

654 Abstracts of scientific papers 2019 AALAS National Meeting

P185 Validation of Equipment Reuse for Germfree Facility Water After several years on hiatus, our stainless steel guinea pig caging Sterilization was reallocated to a different campus only to have our program needs shift and guinea pigs introduced again as a model at our K Patil*, T Bryfogle, A Wagner facility. Faced with a new project without appropriate caging, we turned to the pet store and commercial industries for inspiration. Our University Animal Care, University of Arizona, Tucson, AZ initial solution was open-topped, rugged plastic tubs capable of housing 8-12 adult guinea pigs. The tubs were furnished with plastic Gnotobiotic and germfree (GF/GN) rodent facilities must sterilize all huts, manipulanda, and feeding and watering devices appropriate food and water which is fed to their animals to ensure they for guinea pigs. Our first few cohorts exhibited positive behaviors in continually exclude bacteria, viruses, mold, and fungi from these the tubs such as foraging and chewing on toys. Later in the project, populations. Most commonly, water is sterilized by autoclaving small groups were ordered from a different vendor and displayed more quantities in glass bottles for both isolator and gnotobiotic cage inhibited behaviors. The staff theorized that their inclination to hide systems. These bottles are subsequently submerged in sterilant baths was reserved for when people entered the room. To get a clear before introduction into the GF/GN housing units and so must picture of the guinea pigs’ daily activities, we began behavioral data maintain a strict leak-proof seal. Many facilities use similar style collection. Observations were made via an infant monitoring system bottles which are sold with a rubber bottle cap that forms a leak- at various time points throughout standard work hours of 7:30 am to proof vacuum seal as the bottle cools poststerilization. These caps are 4:00 pm, prior to entering the room for husbandry duties. The sold as “single-use.” We sought to see if it was feasible to reuse these behaviors exhibited during the initial assessment window showed bottle caps to reduce our consumable equipment cost while reluctance in the animal’s willingness to leave the plastic huts maintaining water sterility. We autoclaved 3 bottles/caps as per our regardless of human presence, challenging the original hypothesis standard operating procedures and rather than discard them after a that the animals were only inhibited when staff was present. After single use, reprocessed them 3 additional times. Following each monitoring behaviors in the tubs for 2 wk, the animals were autoclave cycle the three bottle/cap pairs were examined to ensure introduced to a new style of caging, a solid-topped rabbit caging they did not leak and there was no gross damage to the caps that modified to house guinea pigs. We saw immediate positive changes would preclude their continued use. In addition, a sample of the in their behavior, resulting in the need to pause observations until the water from within the bottle was taken and both culture and 16S novelty of the housing dissipated. One month later, following qPCR was performed to ensure sterility. Our tests confirm that this acclimation to the new housing, we resumed tracking behaviors. Our particular style of water bottle cap can be used up to 4 times total data showed a 75% decrease in sleeping and hiding behaviors, rather than thrown out after a single use. We estimate spending coupled with a 4-fold increase in active behaviors such as playing, approximately $150 on bottle caps to supply water to each isolator/ exploring, and eating. The animals continued active behaviors year and reuse as demonstrated here would reduce cost to < $40/ regardless of the presence of staff in the room. Our results isolator/year which could result in significant cost savings especially demonstrate the importance of taking the needs of individual in larger, higher-throughput facilities. Reprocessing could also be animals into account when developing caging standards. In this used as a strategy with confidence, given these findings, if there is instance, shifting the housing from open-topped to a covered caging ever difficulty or delay obtaining restocking bottle cap inventory. system was a positive change as demonstrated by the marked increase in species-normative behaviors regardless of the presence of P186 Do It Yourself: Conversion of Plastic Rabbit Caging to Guinea the staff. Pig Housing P188 Improving Zebrafish Larviculture with Feed-enriched Rotifers K Marshall*, H Wolford, K Kimura, L Martin (Brachionus plicatilis)

Oregon National Primate Research Center, Oregon Health and K Ma*1, K Lertpiriyapong2, AA Adedeji2, C Lieggi2, N Lipman2 Science University West Campus, Beaverton, OR 1Tri-Institutional Training Program in Laboratory Animal Medicine After several years on hiatus, our stainless steel guinea pig caging and Science, New York, NY; 2Center of Comparative Medicine and was reallocated to a different campus only to have our program Pathology, MSK & WCM, New York City, NY needs shift and guinea pigs introduced again as a model at our facility. Faced with a new project without appropriate caging we Larval care and husbandry can pose a significant challenge in assessed surplus rabbit racks to ascertain if they could be used for zebrafish colony management. Larval diet must be of the appropriate guinea pigs. Upon evaluation, we had concerns with the large size and meet the nutritional demands of rapid development while fenestrations for rabbit waste and automatic watering valves fixed in minimally impacting water quality. A diet of live rotifers results in a location too high for a smaller species. Working with internal improved survival and enhanced growth when compared to other fabrication staff, we designed a stainless-steel floor insert, serving to single food source diets (i.e., powdered diets and paramecium). The protect the guinea pigs from injuring themselves in a refuse hole and use of a live diet, however, requires more labor than a processed diet. allow for contact bedding. The insert fits snugly inside the cage and The goal of the study was to maximize rotifer nutrition while is easily removed by equipment sanitation staff for cleaning. To minimizing associated labor. Traditionally, saltwater rotifers overcome the water valve height, water bottles were introduced and (Brachionus plicatilis) are reared in water with a salinity of 15g/L, placed at a height more appropriate for the guinea pigs. We have whereas the salinity in a zebrafish tank is typically <1g/L. It is used this adapted equipment successfully for 2 y, housing 122 unknown if rotifers could be raised at a lower salinity, which may animals to date, with no husbandry-related or clinical concerns, decrease the impact on water quality when the rotifer solution is including any for pododermatitis. Modifying our existing caging has added to the larval tank and reduce osmolar shock to the rotifers. In allowed us to meet the needs of a new species in a cost effective and addition, rotifers are typically fed live algae which have variable environmentally friendly manner. nutrition and can pose a biosecurity risk. We hypothesize that rotifers can be propagated and maintained at a salinity of 5g/L and be P187 Listening to the Animals: Using Behavioral Observations to nutritionally enriched with an irradiated powdered larval diet, rather Assess Species-normative Behaviors in Diverse Housing than algae. Rotifers were placed into culture vessels, food deprived Modalities for 48 h, and subsequently fed either liquid algae, an irradiated powdered larval diet (<50μm) or both at salinities of 5g/L and 15g/L. K Marshall*, H Wolford, K Kimura, L Martin Population sizes and percentage of egg-bearing rotifers were quantified daily until the population density plateaued.Results Oregon National Primate Research Center, Oregon Health and indicate that rotifers appear to thrive equally well at both salinities. Science University West Campus, Beaverton, OR Additionally, rotifers can be maintained successfully on powdered

655 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 larval diet alone at densities of >1000 rotifers/mL. To summarize, testing. Testing was conducted with the user blinded to the products. enriching rotifers with a powdered diet raised at 5g/L can potentially Four different surfaces were tested in two different laboratory spaces reduce labor and biosecurity risks, while providing a nutritious, located in Georgia and California. Wipe 1 improved ATP scores by attractive diet for zebrafish larvae. 85%, wipe 2 improved ATP scores by 19%, and wipe 3 improved by 89%. Based on user feedback, wipe 1 was the easiest to remove from P189 Improving Cagewash Operational Efficiencies through Visual the canister, required only 1 wipe to achieve results, had the best Organizational Systems smell, and the least amount of residue. These results suggest that wipe 1 is the most efficient and easiest to use. Prior to adopting wipe KM McDonald*, CJ Elliott 1 into our cleaning procedures, we would like to conduct additional studies to further evaluate the product. Additional studies could Division of Laboratory Animal Resources, University of Pittsburgh, include environmental testing by culturing surfaces with press plates Pittsburgh, PA and testing of additional surface types.

Our primary housing facility observed a rapid 52.9% increase in P191 Healthy Grief Opportunities and Resources for Laboratory mouse census, and simultaneously modified cage change frequency. Animal Professionals To address the impacts on cagewash operational efficiency, we implemented a color-coded cagewash board. Using employee KL Donelson*, J Wood feedback, we identified several issues: 1) increased throughput demands and scheduling difficulties, 2) inaccurate supply Animal Resource Center, UT Southwestern Medical Center, Dallas, production, 3) nonstandardized caging types created confusion, and TX 4) exhausted storage and set up areas impeded productivity. First, we empowered staff to participate in identifying resolutions, and Managing grief is different for each individual. Developing coping assigned them the responsibility of updating the board for their mechanisms is vitally important to assist in the healing process when assigned areas. To reduce supply circulation, we reorganized the one experiences loss. Working in a field that cares for the health and facility, standardizing cage type by corridor, then equitably wellbeing of animals and where euthanasia is a necessity is distributed cage change across weekdays. We then created a particularly challenging. We outline healthy coping methods, cagewash board with the days of the week listed, and under each activities, and procedural standards that can aid in managing grief at day, the rooms and supplies associated with the change days. Next, your institution specified. A synopsis of talking points will include we implemented a 4-week color coding system, and the room implementing creative enrichment activities, euthanasia scheduling numbers were color coded to indicate supply needs for the color of staff rotations, grief education, and ways to acknowledge and show the week. After implementation, staff conveyed that using the vendor appreciation to our animal cohorts for their contributions to medical names of cage types created confusion. In response, we converted to science. After implementing several of these practices, employees a visual system, using stickers that correlated to a caging type. Next, have vocalized their increased ability to manage their grief in a safe staff stated that supplies were often missing after preparation. In and supportive environment. Based on our observations, response, corridors were color coded, and the associated corridors’ implementing these practices would be recommended for application supply carts were labeled with a large color-coded, autoclavable tag in animal care facilities. indicating the room number. Cagewash staff efficiently consolidated carts with like colors in storage. Our processes further evolved to P192 Centralizing Cage Wash Facilities to Reduce Costs for Repairs improve PI cage supply orders, and to include other scheduled and Replacement of Aged Equipment facility tasks, such washing feed barrels. Due to our success, we are currently implementing a supply receiving board in soiled cagewash, K Jimenez*, FC Hankenson allowing staff to schedule their activities more efficiently. These efforts have improved staff engagement, ensured supply availability, Campus Animal Resources, Michigan State University, Lansing, MI reduced caging circulation by approximately 10%, and increased operational efficiencies. Our institutional animal care program has undergone a rapid growth over the last 5 y, with a rodent census increase of ~25% and P190 Are All Disinfectant Wipes Created Equal? An Evaluation of 3 significant changes in our facilities and strategic operations. Ensuring Different Disinfectant Wipes functional and reliable cage washing equipment as part of the operation is vital to support these programmatic changes. At our DL Thor*1, M Call2, L Roman-Laureano2, KP Storves1 institution, there were cage washers and bottle washers in 6 of our 7 rodent-only vivaria. The existing cage washers were varied in age 1Comparative Medicine, Intuitive Surgical, Norcross, (4=20+ years; 1=15 years; 1=10 years old) and the bottle washers were GA; 2Comparative Medicine, Intuitive Surgical, Sunnyvale, CA similarly aged, with 2 struggling to reach the 180°F temperature requirement. One option considered was to replace all of the Maintaining a clean environment in a research laboratory setting is equipment in each location; however the >$2M price tag was not required not only to produce quality research results, but to protect acceptable due to budget constraints. Given the increased risk of the health and wellbeing of the animals and staff. We frequently use failure of aging equipment, we researched the possibility of disinfectant wipes as part of our cleaning procedures. One of the centralizing our cage wash program and transporting equipment methods we use to evaluate effectiveness of our cleaning procedures from one central location to the outlying facilities. It was determined is adenosine triphosphate (ATP) monitoring. Continuous that while the main rodent housing area was undergoing complete improvement of cleaning processes is a large part of the renovations, we would install 2 high-capacity cage washers and 2 responsibilities of our comparative medicine department. As part of bulk sterilizers for the area and operate this as a central service. Cage this responsibility, we elected to evaluate different types of washers were programmed to run bottle cycles, eliminating the need disinfectant wipes to determine the most efficacious product for our for separate bottle washing equipment in each facility. Once the facility. There are numerous disinfectant wipe products and all have centralized cage washing program was in service, we were able to varying claims on product efficacy and usability. We chose three decommission 3 of the aged cage washers and 4 aged bottle washers. different brands of wipes currently available on the market, each The cost savings on equipment for this project was ~$1M, although it having a different primary active ingredient: wipe 1, hydrogen required investing in an additional cargo truck to separate the peroxide; wipe 2: diisobutylphenoxyethoxyethyl dimethyl benzyl transport of clean and soiled supplies around campus. With ammonium chloride and isopropanol; and wipe 3: citric acid. Using continuous efforts to streamline operations, we have developed a manufacturer guidelines provided on each product, we evaluated process where animal care staff place orders for their cage wash ease of use, user feedback, and efficacy of disinfectant via ATP supplies, cage wash staff fill these orders, and our transportation

656 Abstracts of scientific papers 2019 AALAS National Meeting

team delivers orders to each facility on scheduled days each week. weekly for 11 wk, and glucose tolerance tests (GTT) were performed There were several lessons learned along the way, including ensuring at week 10. Body weights of all mice fed high-fat diet significantly employee involvement in processes that impact their work load, increased over time compared to mice fed with regular diet: final strong communications with the service team for the equipment and average body weights of regular diet + corncob bedding (29.99 ±1.66 continuous review of practices to promote operational improvements g) and regular diet + paper bedding (29.7 ±2.29 g) compared with in the centralized cage wash area. corncob + high-fat diet (43.68 ±3.35 g) and paper bedding + high-fat diet (44.88 ±3.24 g). Blood glucose levels at the end of the GTT were P193 Electronic Animal Health Reporting significantly different between diet types and not between bedding types: regular diet + corncob bedding (99.72 ±14.28 mg/ml), regular KK Clark* diet + paper bedding (88.83 ±12.16 mg/ml), high-fat diet + corncob bedding (205 ±61.83 mg/ml), and 60% high-fat diet + paper bedding Animal Care Unit, University of Kansas, Lawrence, KS (191.17 ±65.96 mg/ml). Similarly, significant differences in the final normalized blood glucose levels were observed in between diet types Laboratory animal departments struggle with finding affordable but not bedding types: regular diet + corncob bedding (1.0 ±0.86) and avenues for medical recordkeeping. Common complications include the regular diet + paper bedding (1.01 ±0.16) or high-fat diet + repetitive data entry, cumbersome communication in decentralized corncob bedding (1.48 ±0.18) and high-fat diet + paper bedding (1.50 animal facilities, and delayed reporting of animal health concerns. ±0.09). In conclusion, housing mice in corncob or paper bedding did Our veterinary staff has worked closely with our information not significantly affect animal model development in metabolism technology (IT) department to develop a low-cost, electronic, real- studies based on the variables measured. Considering its potential time reporting system for health case management and medical benefits, paper bedding can provide a good alternative to the recordkeeping using a commonly available software package already commonly used corncob bedding without compromising research used in many academic and industry settings. Before implementation outcomes. of this system, each health case reported by staff was a 4-step process including 2 paper forms; however, the new electronic health system P195 Can Cross-foster Rederivation Eliminate Murine removes these duplicative steps and requires a 1-time entry. The new Polyomavirus in Steppe Mice (Mus spicilegus) Imported from system is user friendly and allows for real-time communication of France? Oui, Oui! health concerns. Animal care technicians are now able to enter all health-related information into the software, submit this information JR Frohlich, KJ Jones, GW Lawson, M Forsythe* to veterinary staff, while remaining cage side in a decentralized animal facility environment. Veterinary staff then receives an email or Office of Laboratory Animal Care, UC Berkeley, Berkeley, CA text message upon initiation of a new health case. The software allows for designating appropriate treatments by veterinary staff and Murine polyoma virus (MPyV) is a rare DNA viral infection in documentation of treatment/observation completion by husbandry modern laboratory mice, but is known to contaminate transplantable staff. The program records all edits made to a health record and time tumors and cell lines. An investigator imported several breeding stamps those edits so there is no question of documentation timing pairs of wild-derived Steppe mice (Mus spicilegus)from France. While versus a paper record that can be edited at any time. Veterinary staff in the quarantine facility, these mice tested positive on serology for can be notified of a new health case at any time or location, and MPyV, as well as mouse hepatitis virus (MHV) and mouse norovirus husbandry personnel can continue with daily tasks without the (MNV). Culling was not considered a viable option due to the wasted time of exiting the animal facility to get to a phone or precious nature of this species of mice, so these pathogens needed to computer. The program allows animal care technicians to attach be cleared prior to the mice entering into our main colony area. pictures for improved communication to veterinary staff in a Cross-foster rederivation has been used successfully in laboratory decentralized facility environment. Electronically entered data can be mice to clear pathogens such as MNV, Helicobacter, and MHV. easily forwarded to the investigator, improving the speed of However, we could not find anything in the current literature communication and reducing duplicative steps. Taken together, regarding whether or not cross-foster rederivation eliminates MPyV implementation of this new electronic reporting system has increased infection. Furthermore, we could not find any literature to support efficiency through elimination of paper health reporting, simplified whether or not cross-foster rederivation was successful between 2 data entry, increased animal welfare through use of real-time different species of Mus. Following a previously described procedure, reporting and additional capabilities to include pictures, and we cross-fostered newborn M. spicilegus pups (less than 24-h-old) streamlined communication between the veterinary team and onto specific pathogen-free (SPF) outbred Swiss Webster (Crl:CFW) research staff. dams. M. spicilegus pups were weaned at 4 wk, and foster dams were tested via serology. M. spicilegus pups were tested via serology once P194 Effects of Paper and Corncob Bedding in Metabolism Studies they reached 8 wk of age. All serology tests on both foster dams and in Mice cross-fostered M. spicilegus pups were negative for all pathogens, including MPyV, MHV, and MNV. Our results suggest that cross- KA Schultz*1, M Islam2, S Abrishami2, M Varghese2, K Singer2, J foster rederivation can be used successfully across Villano1 different Mus species to eliminate MPyV, MHV, and MNV infection.

1Lab Animal Medicine Unit, University of Michigan, Ann Arbor, P196 Don’t Work Hard, Schedule Smart MI; 2Pediatrics-Endocrinology, University of Michigan, Ann Arbor, MI L Muca*

Reproducibility presents an ongoing challenge in the use of animals Insourcing Solutions/ARCH, Charles River Lab/ Boston Children’s in biomedical research. While the focus has remained on Hospital, Boston, MA experimental designs to achieve reproducibility, aspects of husbandry care can present as confounding variables in animal Scheduling can be difficult and time consuming when no access to model development. We investigated the effects of bedding types on current technologies is available and there is a large staff size and a mouse metabolism studies. We compared a commonly used corncob large program. We were finding it was taking management 2 to 3 h to bedding versus low-dust high-absorbency paper bedding in 6-wk- prepare the schedules for 50 technicians for 2 buildings (1building old C57BL/6J mice. Mice (n=48) were divided into 4 groups based on with 2 two floors and the other building with 3 floors) containing 43 bedding and feed types: 1) regular diet + corncob bedding, 2) regular animal rooms and numerous procedure rooms. The schedule was 6 diet + paper bedding, 3) 60% high-fat diet + corncob bedding, and 4) pages long. The technicians were scheduled for different rooms on 60% high-fat diet + paper bedding. Body weight was monitored different floors making work conditions inefficient. Moreover, the

657 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 technicians were spending extra time just trying to understand the New Zealand White rabbits (Oryctolagus cuniculus) are a social schedules and sometimes missing assignments. Technicians were species, and per regulatory and welfare standards, should be housed responsible for making their own cages for their rooms. This caused in groups or pairs whenever possible. One concern with social conflict over equipment and sometimes too many cages were made housing is the possibility of food hoarding by the dominant rabbit, for 1 room and not enough for another. In an attempt to resolve these preventing the subordinate rabbit(s) adequate access to necessary issues, we reevaluated the schedule to make it simpler and less time resources thus confounding longitudinal age-weight matched consuming. We decided to divide the technicians into floor teams. studies. To ensure that the socially housed rabbits in our large Each floor had their own schedule, which made it easier for the transgenic colony are receiving sufficient nourishment, we performed technicians to visually see their tasks and improved efficiency as they a pilot evaluation of weight trends in addition to Body Condition were now only on 1 floor. Schedules took into account the census, the Scores (BCS) of pair housed juvenile and adult male rabbits. It was difficulty of the room, and technician experience. In addition, each hypothesized that stable rabbit pairs would have weights team had experienced technicians in order to help the less comparable to age-matched single-housed rabbits and the BCS experienced staff. We put white boards in each suite to help would not vary more than 1 on a 1-5 BCS scale. Juvenile weights communication between management and technicians. A person were monitored from weaning age (6 wk) to sexual maturity (15-16 from each team was assigned to staging to make cages and a wk). There were no significant differences in body weights between schedule outlining the number and types of cages and change dates single (n=4) compared with pair-housed rabbits (n=8) at 6 or 15 wk. for each room was made so the technicians in staging knew what was Single-housed male rabbits experienced a 100% gain in weight expected. This increased communication amongst the teams and compared with 115% gain in weight in pair-housed male rabbits (15% created accountability and pride in each teams floor. Now, they were variance) between 6 and 15 wk. BCS and weights of adult rabbits working as a team and not as individuals. With the new schedules in were assessed quarterly over a 1-y period. There were no significant place, management is now spending only 10 min each week on the differences between adult pair-housed rabbits (n=4) or single-housed schedule and technicians are able to understand their daily male rabbits (n=2) over a 9-mo period. Our initial findings indicate responsibilities without missing tasks. We continue to evaluate new pair housing male rabbits most likely does not lead to significant ways to create a lean teamwork environment. variations in weight or body condition in otherwise stable pairs, yet additional data is needed to fully elucidate. These results assist in P197 Work with What Sticks: Failure and Success With Field supporting the notion that social or pair housing of male rabbits Applications of Epoxies that Withstand the Rigors of Autoclave provides potential welfare benefits and should allay the fear that Chambers social housing of male rabbits creates substantial stress leading to significant weight variation or alterations in BCS. Additional research L Antonucci*1,2 and inclusion of a larger sample population is currently in process.

1CMR, Rutgers, New Brunswick, NJ; 2Unconfined System LLC, East P199 Corticosterone and Hematological Monitoring to Assess the Patchogue, NY Effect of Cart Transport on Stress Levels in Mice

With concern from AAALAC about the safety of rack washers and M Ellis, LV Kendall* bulk autoclaves our university, along with a commerical partner, has been testing safety mechanisms. Our original goal was to install an Laboratory Animal Resources, Colorado State, Fort Collins, CO internal fiberoptic safety stop (IFSS). After we started this process our autoclave went through major renovations and the manufacture of Mice used in research are subjected to various types of transportation the new components said finalizing the IFSS would void warranty on including shipping from the vendor and transportation between our autoclave. We completed the process of installing the unit, but buildings at institutions. While shipping from the vendor may did not tie it into the controls. In addition to the IFSS we installed a require up to 2 wk of acclimation, short duration transportation safety gate. This gate will prevent the door from closing and not between buildings and use of the animals does not typically involve allow the machine to run if it is not used properly. Some institutions an acclimation period. Transportation stress can lead to the release of would use a block or cage for the same purpose, but the item can be glucocorticoids, which impacts homeostasis. Therefore, reducing misplaced or not use removing the safety precaution. It is not stress is important from a welfare perspective, and a scientific possible to use to load or unload without using the gate. Anything perspective to reduce confounding factors. The purpose of this study you try to add to an autoclave is a challenge because you cannot was to determine the effect of hand carrying and cart transport of compromise the inner chamber. Adhering items to the wall was the mouse cages on the stress response. CD-1 mice were placed in 1 of 6 only way to accomplish this, but high temperatures and pressure groups, n=10 per group. Group 1 served as the positive control and made most adhesives ineffective. The epoxies were tested first by were mice that were shipped by the vendor and euthanized upon using a mock unit that was placed in the autoclave on a cart. Once arrival (D1). The remaining groups were acclimated for 17 d. Group 2 the epoxy held on the mock unit the epoxy was placed on the IFSS mice were carried on a cart (CC) for 15 min ( 17) and euthanized day and was placed on the wall of our autoclave. While running the 18. Group 3 mice were CC for 15 min (day 17) and euthanized day 31. autoclave some of the epoxy didn’t last for 1 load. Some made it Group 4 mice cages were carried by hand (HC) for 15 min (day 17) through several loads before it failed. Through trial and error some of and euthanized day 18. Group 5 mice were HC for 15 min (day 17) the issues were found to be application errors and through speaking and euthanized day 31. Group 6 mice served as a control and were with the epoxy venders we had more success, but eventual failure not removed from the study room. One group (n=5) was euthanized until the last epoxy. After trying the different epoxies, epoxy D was on day 18 and another group (n=5) was euthanized on day 31. Feces successful on adhering both the IFSS and swing door to our was collected at days 1, 7, 14, 17, 21, and 30 for corticosterone autoclave withstanding pressure and steam through 100s of analysis, and blood was collected at euthanasia for complete blood autoclave cycles (vacuum cycle in a steam heat autoclave), with the counts and serum chemistry. As expected, all mice had the highest autoclave reaching 250 degree F in both 25- and 45-min cycles. corticosterone levels immediately after shipping from the vendor. There was no significant difference in the corticosterone levels P198 Weight Variation Trends in Juvenile and Adult Male Pair- between the transport groups. However, hand carried mice were housed New Zealand White Rabbits (Oryctolagus cuniculus) higher than mice transported by cart or the control group the day following transport. There were no significant hematological changes LA Burlingame*, S Thurston, PA Lester in mice transported by hand or cart compared to the control group at day 18. Similarly, there was no significant difference in blood glucose, Unit for Laboratory Animal Medicine, University of Michigan, Ann AST, ALT, or ALP in mice transported by hand or cart compared to Arbor, MI the control group at day 18. These results demonstrate that mice experience similar stress when transported by hand or a cart, and

658 Abstracts of scientific papers 2019 AALAS National Meeting

that an acclimation period for the short transport may not be Our current online request system along with lockbox pickup that necessary. mimics the human retail pharmacy process can be deployed as a standard method at any institution to gain similar efficiencies. P200 Cost Accounting in a Laboratory Animal Facility—A Simplified Approach P202 Evaluation of the Effect of Bedding and Environmental Enrichment on Mouse Activity Patterns through the Use of C Alaras, LS Bercasio* Automated Home Cage Monitoring

Division of Animal Resources, Emory University, Atlanta, GA, Gabon S Graciano1, M Leblanc4, S Villareal1, F Iannello3, K Mosley2, SC Adams1, M Leblanc*1 Previously, the university’s Division of Animal Resources (DAR) employed a detailed cost accounting system that required intensive 1ARD, Salk Institute, La Jolla, CA; 2Tecniplast USA, West Chester, labor hours to calculate per diem and recharge rates. While DAR was PA; 3Tecniplast SpA, Buguggiate, Italy; 4Biological Sciences, Purdue mostly successful in recovering costs, management decided to University, West Lafayette, IN revamp the process. The goal was to reduce cost accounting effort and time for administrative and technical staff, and provide Automated home cage monitoring of laboratory mice is a new management with a well-grounded framework to manage human technology that provides insights on how microenvironment, and financial resources. The legacy system used an animal census husbandry practices, and enrichment may affect animal welfare and database and an outdated database platform which were both behavior as well as research variables. We investigated how different phased out in 2018. The process involved multiple handling of the types of bedding and environmental enrichment influenced activity same data, subjecting data to human errors. Self-reported timecards levels and patterns of mice housed in individually ventilated cages drove the labor cost component of per diem rates, and categorization (IVC). In a first experiment, adult female Swiss Webster (SW) mice of nonlabor costs consumed significant time due to multiple coding (3/cage) were housed in IVC under 5 different bedding conditions and entry, along with disparate coding combinations requiring (soft cellulose bedding, corn cob bedding, paper chip bedding, cob/ conversion. All these resulted to significant delays in rates paper squares, and aspirated wood chips, n=8 cages/condition) with calculation. DAR approached the project as a business process standardized husbandry and enrichment practices. Heatmaps and redesign and devised a project plan, a business requirements activity levels in the entire cage as well as in the corners and the document, and a communications strategy; calculated rates using center were monitored for 1 mo. Overall, mice housed on cob/paper principles and methodologies patterned after updated practices squares bedding showed the highest activity followed by aspirated developed and used by peer universities; and applied a formula to wood chips, corn cob bedding, paper chip bedding, and soft cellulose simplify cost categorization between per diem and special service bedding. Mice housed on soft cellulose bedding displayed recharges. DAR also conducted a baseline time and motion study to significantly less activity and spent less time exploring cage corners evaluate reasonableness of existing relative quotient factors used to as compared to mice held on cob/paper squares. In a second allocate expenses to cost centers. DAR then compared results with experiment, adult female SW mice (3/cage) were housed in IVC current rates derived from the legacy system. Shifting from the cages with corn cob bedding under standard husbandry practices for conventional cost accounting process to the simplified methodology 1 month but were provided different environmental enrichment resulted in significant labor cost savings of $50,880, as well as a devices (none, cotton fiber squares, commercially available paper- reduction in rates calculation and preparation time. For example, hut, a plastic hut, commercially available nesting material pucks, and there was decrease of 2,189 annual labor hours due to the plastic tube, n=8 cages/condition). Overall, mice housed with plastic replacement of 2 senior accountants with 1 financial analyst. DAR huts showed the highest activity followed by no enrichment, cotton eliminated manual self-recording and preparation of monthly fiber squares, paper-huts, plastic tubes, and nesting material pucks. timecards plus document submission previously required from all Activity levels were significantly higher in cages with plastic huts as 100 divisional staff members, as well as the coding non labor expense compared to cages with paper-huts, plastic tubes, and nesting transactions, compiling and processing of labor time submissions, material pucks, especially during the night phase. Activity levels in and entering labor and nonlabor expenses onto 2 database platforms. cages without enrichment and with Nestlets were also significantly There was also consolidation of mouse cost centers from 11 to 5. higher than cages with nesting material pucks. Mice housed with plastic huts spent more time in the center of the cage while mice P201 Lean Approach Delivering Efficiency and Traceability for housed with cotton fiber squares spent the most time exploring the Centralized Drug Dispensation cage corners. These data demonstrate that automated home cage monitoring can be used to assess the effects of cage environment and M Connell*, A Cummins, C Doerning, S Tummala husbandry practices on mouse behavior. Additionally, results suggest that bedding and enrichment types can significantly affect levels and Division of Veterinary Services, Cincinnati Children’s Hospital patterns of activity of laboratory mice. Medical Center, Cincinnati, OH P203 Improved Husbandry Practices Have Decreased Spontaneous The majority of anesthetic and analgesic agents used in research are Morbidity and Mortality in an NSG Mouse Colony controlled substances. Distribution of these substances to research investigators is integral for providing appropriate anesthesia and M Gallacher*2, V Ospina2, M DuPont1, K Bell2, C Hoover3 analgesia to meet welfare and regulatory requirements, as well as generating reproducible scientific data. Management of the 1Oncology Bioscience In Vivo, Early TDE Discovery, Oncology R&D, distribution process for controlled drugs, however, creates AstraZeneca, Boston, MA; 2Animal Sciences and Technologies, operational challenges in terms of staff availability, logistics, Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, regulatory compliance with protocol and DEA mandates, and timely Boston, MA; 3Pathology, Clinical Pharmacology and Safety Sciences, access. The lean concept of value stream mapping has been used to R&D, AstraZeneca, Boston, MA link together 2 key elements necessary to standardize and streamline drug distribution and create an improved process. An online request NSG mice (NOD.Cg-Prkdcscid Il2rgtm1Wjl/Sz) are severely system has been developed to generate a standardized request and a immunocompromised mice that are used in the development of secure lockbox pickup system has been implemented to manage the xenograft and patient derived tumor models to research novel chain of control for controlled drugs. These processes have led to an medicines intended to treat various human cancers. In 2018, we improved documentation system with tighter regulatory control, unexpectedly experienced high morbidity and mortality in an NSG decreased ad hoc drug distribution, and a substantial improvement mouse colony housed in barrier conditions. Microscopic examination in convenience and value for both veterinary and research personnel. of the gastrointestinal tract collected from clinically ill NSG mice

659 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 revealed heavy bacterial growth and mucosal atrophy. Aerobic and The addition of new research personnel drives the need to provide anaerobic cultures, and polymerase chain reaction (PCR) for vivarium access. However, because of the magnitude and complexity clostridial toxins did not identify any opportunistic pathogens in of the work involved there is minimal impetus to regularly recertify animals displaying clinical signs consistent with a diagnosis of and remove user access from staff for which it is no longer required. microbial dysbiosis. To decrease mortality, we decided to depopulate This results in users that maintain access indefinitely. Initial access the remaining NSG mice, sterilize the barrier facility with vaporized requests typically require verification that medical, training, and hydrogen peroxide, and adopt updated husbandry, traffic, and security requirements are complete. However, it is often difficult to handling practices for studies using immunocompromised mice. ensure that the requirements continue to be met over time. These Facility sterilization was confirmed via biological indicator analysis limitations may result in compliance issues, safety risks, and security for G. stearothermophilius. Revised traffic patterns, reduced access to threats. In response, we have developed a digital system, that NSG holding rooms, and frequent disassembly and cleaning of integrates data from multiple sources to ensure that vivarium access biosafety cabinets were implemented following confirmation of the requirements are continuously met. Following an initial request, the facility sterilization. To monitor for any recurrence of dysbiosis, we system confirms that there is a legitimate need for vivarium access by established a weekly sentinel program with NSG mice which consists verifying appropriate institutional employment/appointment status of histopathological examination of all segments of the and cross-referencing that the user is listed on an active animal gastrointestinal tract. Samples of small intestine and feces are frozen protocol. The system then proceeds to verify that the user’s training for additional diagnostics as needed. Approximately 1,500 NSG mice and occupational health requirements have been met via automated have been housed following the barrier sterilization. Microscopic database queries. Following requirement verification, a notification is examination of NSG mice has not shown any signs of dysbiosis and sent to security personnel to grant access to specific areas based on an the NSG colony has not had any cases of unexplained death through established algorithm. Once access has been granted the system 14 wk. Using the NSG sentinels has provided confidence in the continuously monitors the requirements for thousands of user depopulate-sterilize-repopulate procedure performed to halt the accounts from multiple institutions. Anomalies or deficiencies trigger unexpected high morbidity and mortality. Furthermore, microscopic a user notification with instructions to correct. After a grace period, review of NSG sentinel gut has validated the effectiveness of the inaction results in withdrawal of access. In a large institution with handling and husbandry refinements put in place to limit dysbiosis hundreds of requests every month, this platform has streamlined the in the NSG colony. process, reducing the time and employee effort required to provide P204 A Novel Cage Card System for Clinical Management of access while detecting gaps and eliminating human error. The Phenotypic or Model-induced Rectal Prolapse in Mice (Mus system’s intuitive and user-friendly interface has also improved user musculus) satisfaction with the ability to easily view information and track the progress of a request in real time. Although the system was built to M Lundberg*, S Lewis address the specific needs of the participating institutions, the framework can be customized for use by other organizations. Animal Resource Center, UT Southwestern Medical Center, McKinney, TX P206 Transitioning to a New Compliance and Animal Facility Administration Software At our institution, there are many mouse (Mus musculus) models (Inflammatory Bowel Disease) or research related-phenotypes B Trotter, M Sketers*, H Tekie, N Montero, D Munoz, J Sketers, M (truncated anatomy) that result in large colonies of mice with rectal Montero, L Bell, DL Fong, CA Manuel, L Richardson, C Russell, JK prolapse. Typical case management starts with assessing the mouse, Leszczynski then recording the exam on a carbon paper health report; one is left cage side, and the other kept for physical and digital records. This OLAR, University Of Colorado, Aurora, CO reporting has become burdensome because of the time spent on each individual case. To circumvent this issue, while still providing An effective compliance and animal facility administration software appropriate clinical care, a new system was developed for colonies of (CAFA) can provide significant operational efficiencies and cost mice with rectal prolapse due to research–related or phenotypic savings for animal research facilities. Our institution recently issues. The card system developed operates as follows: once a rectal implemented new software. Our centrally managed animal resource prolapse is observed and reported by the animal technician, a program has approximately 23,000 cages distributed within 5 veterinary staff member places a laminated rectal prolapse card on facilities, over 500 active IACUC protocols, and over 350 principal the cage. The veterinary technician fills out one side with the rectal investigators with approximately 1,200 research support staff. The prolapse grade (1-3 or mild to severe), the date it was last seen by the transition impacted all aspects of IACUC and IBC protocol veterinary technician, and the clinical case number. Once a grade 2 management, financial, and animal facility management. IACUC, rectal prolapse is observed by the veterinary technician, the accounting, and animal facility management modules were husbandry staff is trained to change the standard bedding (wood converted from 1 proprietary software to an entirely new platform. chip or corn cob) to compacted cotton bedding. At this point, the System migration was challenged by the need for complete veterinary technician will also start the mouse on topical antibiotic replacement within 8 mo from the time of signing the contract to full ointment to keep the prolapsed tissue moist and free of infection. implementation. In order to prepare for the migration, a team of 3 Once a grade 3 prolapse is reached, the card is flipped and on the temporary employees and 7 existing employees developed data lists, opposite side is a request for the lab to euthanize along with the date along with methods to validate the new platform. Once the system and time of deadline. An email is sent to the investigative staff to was delivered and validated, approximately 30 employees consisting alert them to euthanize their mice. Cards are checked by veterinary of facility staff, administrative staff, veterinarians, and IACUC staff technicians once per week. Implementation of this system has transferred all data and protocols into the new system manually over benefited both investigative and veterinary technical staff by a 6-wk period. Development of a variety of training materials were increasing cage-side communication and decreasing excess an integral component to the effective training of staff on facility paperwork. management modules and researchers on IACUC modules, as well as financial reports. The successful CAFA data migration resulted in P205 A Digital Locksmith: Development of an Integrated and more accurate census and billing allowing a reduction in labor cost Automated Facility Access Management System with an approximate value of $120,000 annually. Integration of the IBC allowed for the IBC and IACUC protocols to be linked, helping M Eckstein*1,2 with compliance. Investigators can now review their cage census and invoices at any time and correct discrepancies as well as create and 1CCMP, Weill Cornell Medicine, New York, NY; 2CCMP, Memorial amend their protocols with ease. This has allowed billing processing Sloan Kettering, New York, NY time to be reduced by approximately 4 d. Researchers’ feedback

660 Abstracts of scientific papers 2019 AALAS National Meeting

continues to be positive, both regarding the IACUC protocol process, animals, indicating environmental monitoring is useful to evaluate as well as the animal facility management modules. This shows that risk of IVC cage contamination. This data can support decisions it is possible to successfully transition a large academic animal about sample numbers for conventional animal health monitoring resource program to a new software in a limited timeframe. based on actual environmental risk. We believe this monitoring may have applicability to a variety of facility types. Environmental P207 What Evil Lurks in Your Vivarium? monitoring is a relatively easy and inexpensive technique with benefits for rodent production. P Sharp, I Faseeh* P209 Evaluation of Rodent Feed Usage during Cage Changing Animal Resources Centre, Murdoch, Australia R Gerhardt* The laboratory animal holding facility, or vivarium, is a purpose-built building to accommodate research animals and exclude deleterious Animal Resources, Virginia Commonwealth University, Richmond, agents. To appropriately maintain SPF mice and rats, rodent Virginia, VA vivariums should be well-planned, well-designed, and well- constructed. Appropriately designed and built doors are an We sought to determine if reusing gamma irradiated feed remaining important part of the contemporary vivarium. As per the Guide for the in the hopper at the time of cage change would reduce overall costs Care and Use of Laboratory Animals, doors should fit tightly within while not impacting animal health. Previously, we disposed of any their frames, and both doors and frames should be appropriately remaining feed in the hopper every 14 d when the cage was changed. sealed to prevent vermin entry or harborage. Various door types may For this study, during 1 cage change, feed remaining in the hopper be used; some are solid and others hollow. The top and the bottom of was transferred to the new cage and additional new feed placed on the door is rarely inspected to ensure that the integrity of a hollow top to ensure there was adequate levels. All feed was discarded at the door has not been compromised. Recently we renovated a barrier next cage change to ensure that food pellets would not approach area with hollow aluminum doors. Upon removing and opening the expiration. Thirty female CD-1 mice were separated into 6 cages, 5 hollow doors, we found the void in the doors were full of dust, but per cage. Three cages received a new hopper of food at every change no evidence of vermin. It was never verified when the doors were (14 d). Three cages would have the remaining feed transferred from installed (a decade or more ago) that the seal holding the 2 halves of the old cage and additional feed added to fill the hopper. Every 28 d the doors together were in fact airtight. Preliminary microbiological these animals would receive all new feed. The mice were weighed investigation of the dust found bacteria (Pasteurella weekly during cage changing and food and water checks. The food pneumotropica, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella was also weighed at this time to see if their intake was affected. There oxytoca, Klebsiella pneumoniae) some of which could be undesirable in was no statistical difference between the 2 groups, although the SPF rodent colonies. No viral or parasitic agents were detected. Some group that had the feed transferred every other change gained more of the bacterial agents identified in the hollow space were also weight. Overall, transferring feed between cages did not show a detected in animals in the facility. This investigation highlights the negative impact on weight gain. By transferring the feed between importance of the proper construction of vivarium doors and the cages we determined a substantial reduction in the amount of feed risks associated with all hollow spaces within the vivarium. Hollow needed, which also serves to reduces costs to the animal program. doors or other spaces inside the vivaria require strong reconsideration not only for their potential to harbor vermin, but for P210 Introduction of the Chinchilla (Chinchilla lanigera) into an their now demonstrated ability to harbor infectious agents. Animal Care and Use Program at a GLP-compliant Facility

P208 Translating Best Practices from Cleanroom Manufacturing: RA LaFleur*, K Arthur, MJ Williams-Fritze Environmental Monitoring of IVC Production Rooms to Evaluate Risk of Bacterial Contamination CBSET Inc, Lexington, MA

PL Roesch*, J Brown, J Olson We recently introduced chinchillas into our program to support auditory studies. Chinchillas are often used for auditory research due Molecular and Diagnostic Analysis, Taconic Biosciences, Rensselaer, to their accessible tympanic bullae and similarities to the human NY hearing range. Here we describe the steps taken to integrate chinchillas into our program. Since the well-known commercial Cleanrooms are specialized facilities that use high-efficiency vendors do not sell chinchillas, we needed to identify a vendor from particulate air (HEPA) filtration to maintain low particulate levels. which to source chinchillas. After identifying a vendor, we worked They are commonly used for pharmaceutical and computer chip with a commercial laboratory to create a custom chinchilla health manufacturing. Cleanrooms employ environmental monitoring screening panel that was employed prior to animal importation. programs to survey microorganisms and particles, which might Since the chinchilla vendor did not offer commercial transport, we provide useful information to supplement a conventional animal identified a courier service to deliver the animals to our facility to health monitoring program. Our SOPF rodent production rooms ensure compliance with the Animal Welfare Regulations. The primarily use individually ventilated cages (IVCs), which provide chinchillas were housed in modified rabbit caging with custom-cut good biosecurity but are difficult to monitor for bacterial plexiglass solid flooring placed over a portion of the existing cage contaminants. Our hypothesis was that monitoring of bacteria in the floor. Caging was setup with vertical space to encourage species- production room environment could serve the following purposes: to specific behaviors such as jumping and climbing. We quickly learned assess the risk for contamination of IVC cages from excluded that chinchillas can easily escape through the small spaces in a subset organisms, to evaluate how well our PPE and processes restrict entry of our housing banks, which limited our housing options. Animals of microorganisms into the production room, and to identify facility/ were socially housed in same-sex pairs or trios. A variety of food and procedural failures. We instituted an environmental monitoring environmental enrichment was provided, as well as frequent program that consists of monthly room air sampling using a interaction with the staff. As a GLP-compliant lab, we also updated microbial air sampler. Approximately 10 samples are collected per all of our relevant standard operating procedures to include room. Air samples are cultured and bacteria are identified to the chinchillas and documented training of our staff on behavior, species level. Results from up to 12 mo of monitoring indicate that husbandry, and handling. We highlight several of the research uses of our IVC production rooms have surprisingly low levels of bacterial our chinchillas, including auditory brainstem response testing (ABR), contaminants, thus supporting the effectiveness of our PPE and transbulla dosing, and cochlear imaging. processes. We found good correlation between microorganisms found in the production room environment and those found in our

661 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P211 Weight Tracking as a Tool to Predict Parturition in the scheduling via an institutional cage change benchmark has aided our Common Marmoset (Callithrix jacchus) facility to streamline scheduling processes and overall personnel accountability. This is a fair and transparent approach to normalizing SL Bokor*2, EB Issa2, CL Winnicker1, R Ober1 and evenly distributing workloads for each week’s schedule. Our standardization and simplification of the schedule has proven 1Institute of Comparative Medicine, Columbia University, New York successful both for the employee and the facility. City, NY; 2Department of Neuroscience, Columbia University, New York, NY P213 The Effect of Ultrasonic Fogging with Peracetic Acid and Hydrogen Peroxide on Rat Pinworm Egg Viability The common marmoset (Callithrix jacchus) is known as the most fertile anthropoid primate due to its multiparous tendencies, SM Jaber*1,2, EA Jaskolski1, D O’Connell3,1 relatively short gestation (mean=143-144 d), and absence of a lactational anoestrus period. Similar to humans, marmosets have a 1Animal Medicine, University of Massachusetts Medical School, 28-d ovarian cycle in which copulation can occur throughout. Unlike Worcester, MA; 2Pathology, University of Massachusetts Medical other primate species, however, female marmosets do not menstruate School, Worcester, MA; 3Abbvie, Worcester, MA or display external characteristics of ovulation. Additionally, most of the endocrinologic tests used for pregnancy detection in humans and Syphacia pinworms are a frequent parasitic pathogen encountered in Old World primates are unreliable in marmosets. High-resolution rodent vivaria. Their presence induces physiological responses that ultrasound equipment as well as crown-rump measurements may can complicate research. Pinworm eggs can be found in many parts assist in delivery date estimates, but can be inaccurate and cause of the infested facilities, some of which are difficult to clean because distress to the animal. In order to properly prepare resources in the of hard to reach spaces or sensitive equipment. Syphacia eggs are case of triplets where significant intervention in assisting with infant- also notoriously resistant to disinfection. We evaluated the ability of rearing would be necessary, it is critical to be able to predict reliable an ultrasonic fogging device that uses low concentrations of peracetic delivery dates. We devised a simple, noninvasive, and inexpensive acid and hydrogen peroxide on killing rat pinworm eggs. Rat method to track prenatal growth and estimate delivery date through pinworm eggs were collected from privately owned rats using consistent weighing of a breeding female marmoset. Animals are double-sided cellophane tape. They were allocated to control, trained to enter a transport box in order to voluntarily participate in ultrasonic fogging, or soaking in liquid chlorine dioxide groups prior the weighing process. Females marmosets are weighed for several to incubation in hatching media. Viability was assessed by opening of weeks before the mating process begins to establish a baseline, and the egg shell operculum detected via light microscopy. Percent then are continuously weighed 1-2 times per week after pairing and hatching between control and ultrasonic fogging groups were similar during suspected pregnancy. While a pregnant dam’s weight has (78% and 80%, respectively) and was less in the chlorine dioxide minimal change from baseline for the first and second trimester, a group (11%). We concluded that at the concentration and dwell time statistically significant inflection point of 30-50g, depending on litter used for the study, the use of ultrasonic fogging with low size, occurs at approximately 103 d, from which it is possible to concentration peracetic acid and hydrogen peroxide is not effective in accurately predict the date of delivery for a typical gestation rendering rat pinworm eggs nonviable. The use of double-sided tape schedule. Using operant conditioning to elicit cooperation from the for collection, evaluation, and incubation in hatching media animals, marmoset parturition can be predicted accurately based on increased the efficiency of experiments compared to previously noninvasive, low-stress regular weighing of female marmosets. reported techniques that involved inverting tape between evaluation of egg number and exposure to hatching media. P212 Standardizing and Simplifying Staff Schedules P214 An Evidence-based Approach to Reducing Potable Water RA Franceschi*, M Rosenbaum Waste in a Laboratory Animal Facility

Biological resource center, National Jewish Health, Denver, CO S Hashway*1, M Suckow2

Creating a fair schedule, recognizing individual strengths and 1Research Animal Resources, University of Minnesota, Minneapolis, autonomy, while promoting transparency with the vivarium’s MN; 2University of Kentucky, Lexington, KY personnel are common challenges for animal care managers. Our facility implemented a benchmark of 50 cage changes per hour Laboratory animal facilities are resource-intensive operations, due in (50ccph), using the traditional time and motion studies with current large part to the need for appropriate contamination control. Over staff, and provide a daily allotment of cages for each animal care the past decade, there has been an increased focus on sustainable technician using principles introduced by lean management strategies to conserve resources within animal vivaria. One concepts. Managing a benchmark of 50 ccph has enabled us to indispensable contributor to resource consumption is the provision of eliminate the traditional time and motion studies that are often used potable water for rodents. The Guide states that “animals should have when developing schedules for the care staff. Each preceding week, access to potable, uncontaminated drinking water,» but does not the care staff provides estimated time needed for technical expand on those requirements, leaving the definition of “potable” responsibilities along with the number of cages they will be changing and “uncontaminated” open for interpretation based on in their assigned area for the following week’s workload. In turn, requirements established by municipalities and the needs of the management provides a specified daily cage allotment to minimize research itself. While industry standards exist for the frequency of wasted cage preparation and allow for accurate workload replacement of used rodent water bottles, no such standard has been scheduling. Using the 50ccph benchmark, all anticipated workload determined for filled rodent water bottles that are stored for later hours are normalized into cage numbers. For example, 5 h of use. Potential contamination sources limiting the shelf life of stored technical responsibilities are converted to 250 cages. If a technician water bottles include microbial contamination and leaching of has 1,000 cages to change and 5 h of technical work for the week, this inorganic compounds from water bottle stoppers. In an effort to results in a total of 1250 cages or 250 cages per day (in a 5-d week). decrease waste associated with replacing unused rodent water Thus, management expects 5 h of scheduled work per day. Each bottles, this study evaluated microbial and inorganic contamination technician’s additional time can be used for ancillary assignments in rodent water bottles stored for up to 28 d. Autoclaved and throughout the facility. Staff knowing their weekly/daily tasks that nonautoclaved water bottles were stored in filter-topped, static need to be completed allows for individual autonomy of their polycarbonate rat cages measuring 10.5” x 19” x 8», with 20 bottles assigned schedule. Allowing staff to manage their own schedule per cage. Water samples were submitted for bacterial culture and provides flexibility and ownership, allowing control of day to day inorganic mineral analysis at 0, 7, 14, 21, and 28 d post-filling. To assignments. In summary, providing daily cage allotments and identify potential biofilm, sipper tube culture swabs and water

662 Abstracts of scientific papers 2019 AALAS National Meeting

expressed through the sipper tubes were also submitted for bacterial Managers have made significant progress in continuous cultures at d14 and d21. At all timepoints, inorganic mineral content improvement at the institutions which have implemented some or all was well below national drinking water maximum contaminant of the tools. Importantly, employees report less frustration with daily levels established by the Environmental Protection Agency. While all tasks, and increased feeling of purpose that the work they do matters water cultures were negative for bacterial growth through d 21, in the organization. Quantifiable improvements achieved using the cultures from sipper tube swabs and water expressed through sipper basic tools and techniques outlined in the Manager’s Toolkit include tubes revealed bacterial growth from several bottles starting at d 21. 1) Improved customer service and customer relations; 2) better Based on these results, our department has extended the limit for performance with the AAALAC accreditation process; 3) more storage of unused rodent water bottles from 7 to 14 d. This study efficient use of human resources (FTEs); 4) increased quality of represents an evidence-based approach to reducing potable water animal care; 5) improvements in space utilization and overhead waste in a laboratory animal facility. distribution; 6) reductions in operational costs and maximization of limited budgets; and 7) improvements in overall staff morale. As lean P215 Establishing a Colony of African Spiny Mice (Acomys management becomes the preferred method for achieving cahirinus): Maintaining and Breeding an Unconventional species operational excellence, the tools, techniques, and assistance offered in a Traditional Animal Vivarium by the toolkit can equip managers with the confidence and capability to improve operations, efficiency, quality, and create an SM Soprano*, EK Daugherity organizational culture of excellence.

Center for Animal Resources and Education, Cornell University, P217 Scoop It or Count It: Which Feeding Method Is More Cortland, NY Efficient in Saving You Time Overall?

African spiny mice (Acomys cahirinus) have become more common as S Phillips* research models in recent years because of unique biological characteristics. Primarily, spiny mice are used to study tissue Division of Research, University of Houston, Houston, TX regeneration due to a weak skin phenotype and faster wound healing as compared to other rodents. Our breeding and colony management Nonhuman primate cages (NHP) are cleaned daily to prevent personnel were recently tasked with establishing adequate housing, a accumulation of debris in accordance with the USDA Animal Welfare husbandry program, and a breeding colony of African spiny mice. Regulations. In our primate facility, the majority of husbandry Since the spiny mouse is a relatively unconventional lab animal technician time is dedicated to feeding primates and daily cleaning of species that was not previously housed at our institution, we were cages and rooms. Cage debris that must be removed includes faced with unique challenges related to establishing a successful uneaten biscuits. We observed that a substantial amount of time was colony in a traditional biomedical vivaria. We present specific required to break down uneaten biscuits with a hot water hose. Our challenges, environmental modifications, novel housing, and current practice is to feed primates with a standard scoop for enrichment used while establishing, maintaining, and breeding a juveniles, adult males, and adult females. This method is consistent colony of spiny mice. Changes implemented include the addition of and simple, but this practice can lead to excess biscuits remaining in larger cages to accommodate harem housing, modification of the cage or obesity. An alternative practice is creating individual temperature and humidity that would closely mimic environmental biscuit counts for each primate based on biscuit consumption conditions in Africa, and the addition of branches and cave structures analysis and veterinarian consult. Potential benefits include optimal to model the natural environment. This resulted in improved body condition and reduced amount of leftover biscuits. However, breeding success, increased natural behaviors, and a decreased time spent counting biscuits may offset time saved cleaning cages. number of clinical cases. Finally, we outline future plans for We chose to compare the 2 feeding practices’ impact on time required maintaining this colony during experimental manipulation. The to clean the cage and provide biscuits. We hypothesize that approaches outlined will serve as a resource for animal care and use individual biscuit counts will be the most time efficient method of programs required to house and breed species novel to their vivaria. providing feed and cleaning primate cages. Comparisons of mean time required to clean cages and provide biscuits when the different P216 Lean Management: A Leader’s Toolkit for Achieving feeding practices are used will determine the most efficient practice. Operational Excellence This evaluation will optimize the use of our husbandry technician’s time. SM Kirchain*1,2, J Booth3,2, G Cronin4,2, DM Jarrell4,2 P218 Introduction of A Web-based Novel Laboratory Animal 1CCM, Brigham and Women’s Hospital, Arlington, MA; 2Vivarium Management System Operational Excellence Network, Charlestown, MA; 3Penn State College of Medicine, Hershey, PA; 4Center for Comparative Medicine, Y Hu1, X Wu2, Y Tao1, S Kong*1 Massachusetts General Hospital, Boston, MA 1Peking University First Hospital, Beijing, China; 2Heng He Yi Shang Lean management has become a popular model for efficient Technology (Beijing) Limited Company, Beijing, China vivarium operations at biomedical research organizations around the world. While lean Management is a widely known concept, the steps The goal of this new system was to improve the work performance in needed to begin an effective lean program are not so obvious. Here a laboratory animal facility through enhancing the communication we present our development of a Leader’s Toolkit, providing between lab staffs and researchers (users). The database in this foundational tools and techniques that can be easily implemented to system consists of research user account information including an ID help managers start achieving a culture of empowerment, number and other useful information, as well as records of all animal engagement, and continuous improvement in the vivarium setting. information, such as arrival time, age, breeding condition, genetic The toolkit was created with the goal to facilitate 1) an orderly background, cage number. The animal cage is the key part in this workplace; 2) scientific approach to problem-solving in the system, and all information of animals listed above is linked to each workplace; and 3) response to organizational challenges. The tool kit cage and converted to a QR code printed on each cage label. A cage is comprised of 11 tools: 5S scoring sheets, 5 “whys” sheet, fishbone label can be printed on a self-service terminal printer by any user Analysis, A3 problem-solving Worksheet, impact-effort matrix, go with a valid account ID. The database also contains data collected and see/gemba form, downtimw, standard work template, and statistically summarized, and with the evaluation of use of the stakeholder analysis, plus-delta template, and control chart template. facility, as well as resources information, such as available cage Visual examples of each of the tools are depicted, with a brief numbers, equipment, and technicians. Research users can place order description and tutorial on their use in applicable situations. or make an appointment after viewing this information. To protect

663 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 the individual privacy and system safety, the system holds 2 substrates and environmental enrichment used in studies where FBG permission authorities: administrator and research users with is measured, in order to prevent confounding results. different levels of function. This system is Wechat and internet accessible for ease of use. Facility staff can use mobile phones to P221 Tarps Used as Visual Barriers Decrease Occurrences of collect census and other data, and research users can use mobile Aggressive Conflicts in Baboons phones to update data or apply for experiment resources anywhere at any time. In addition, facility staff can have rapid communication TL Stevens*, J Cain, J Daniel with researchers through the application regarding changes in animal health conditions, and researchers can request special care for specific Comparative Medicine, The University of Oklahoma Health Sciences animals through the application. By using open source software such Center, Oklahoma City, OK as mysql and tomcat etc, this system meets the facility requirements. Baboons have a biological need to socialize with conspecifics, but at P219 Evaluating the Appropriate Sanitization Frequency for times they may require privacy and a means to avoid conflict with Rodent Caging Accessories others. In some instances just breaking visual contact with other baboons is all that is needed to diffuse a hostile situation. Nonhuman S Poweska*, M Bellamy, J Maher, M Simkins primate colonies housed in research settings are manipulated frequently due to research protocols, for breeding, and for veterinary Comparative Medicine, Pfizer, Pearl River, NY procedures. These manipulations often cause a shift in the strict social hierarchy leading to restructuring of the troop, which is not The purpose of the study was to determine the appropriate always pleasant. Providing visual barriers inside of corral areas, such sanitization frequency of mouse caging accessories. We conducted a as barrels and panels is widely used in primate facilities to allow study to evaluate extending the frequency of mouse cage accessory subordinate animals a chance to avoid dominant/aggressive conflict sanitization from 14 to 28 d. The study was conducted using CD-1 between individuals in the same troop. However, the issue we came female mice housed 5 per cage. There were 10 cages of animals with 1 across was neighboring baboon colonies becoming involved and of the cages being a sham (no animals in cage). The mice were even initiating disputes in other corrals. Aggressive posturing and housed in IVC cages with ¼” irradiated corn cob bedding nd one gestures from across the room can cause entire troops to become irradiated nest. The mice were fed pelleted rodent diet. The rodent agitated, causing redirection of aggression to conspecifics. Our caging accessories sampled were mouse igloos and wire bar feeders. institution recently found itself observing this redirected aggression The sampling of the accessories was conducted on day 0, 14, and 28. first-hand while consolidating our colony of 3 existing troops into 2 Samples were collected and shipped on ice to an independent lab, troops. Baboons from the divided group were showing increased incubated for 48 h, and read. The swabs were analyzed in house with frustration and stress when they would witness their former cohorts the ATP tester. The statistical results of the of 95% confidence interval involved in an even minor altercation, such as chasing. This behavior indicated that day 14 and 28 are equivalent for both the RODAC would have a domino effect eventually leading to all corrals plates and ATP testing. No colonies were observed on the becoming agitated and aggressive, sometimes leading to injuries that MacConkey II plates therefore statistical tests were irrelevant. In require medical attention. We found an easy solution to provide a conclusion, we can extend the sanitation period from 14 to 28 d quick and effective visual barrier between large corral housing, as because the results of the RODAC, McConkey II, and Relative Light well as a portable, smaller unit for noncolony housed baboons. A Units (RLU) demonstrated equivalency between 14 and 28 d. simple design of using a large tarp affixed to a rod and hooks, has had a huge, positive impact on our baboon colony. After adding the P220 Effects of Bedding Substrates and Environmental Enrichment visual barrier between opposing groups we had over a 60% decrease on Fasting Blood Glucose in Mice in aggressive encounters within a 6-mo period. Overall, we found that incorporating these large, colonywide visual barriers are J Kropik1, SY Kondo*1, SE Seifried2, M Wong1 beneficial for occluding negative visual interactions between neighboring troops, leading to a decrease in stress and aggressive 1Animal and Veterinary Services, University of Hawaii, Honolulu, conflicts between our baboon colonies. HI; 2Cell and Molecular Biology, University of Hawaii, Honolulu, HI P222 Bringing Creativity and Variety to Our Research Monkey’s Thirty 8-wk-old C57BL/6 female mice were randomly distributed to Environments 6 cages (5 mice per cage). Each cage was subjected to 3 weeklong treatments replicated twice for each treatment over a 6-wk period. TJ Chase* Treatments consisted of either 1) hardwood bedding, 2) hardwood bedding plus an enrichment item made with food-grade adhesives, Department of Comparative Medicine, Oregon National Primate or 3) corn cob bedding. At the beginning of each week, baseline Research Center, Beaverton, OR nonfasting blood glucose was measured, and at the end of each week, following a 14-hr fast, fasting blood glucose (FBG) was Beyond receiving the required nutrients like protein and vitamins, measured for each mouse. In addition, mice were weighed twice a novelty and foraging are necessary to ensure healthy breeding week. By randomizing each cage’s order of treatment, any effects due groups of monkeys at our research institutions. The husbandry team to the age of the mice or order-of-exposure were factored out. The is part of the comprehensive enrichment program that provides data was subjected to statistical analysis using notched box plots and enrichment to ~5000 nonhuman primates housed in both indoor and a series of tests. The results demonstrated that mice housed on corn outdoor enclosures. The husbandry team uses surplus seasonal cob bedding presented a modest but statistically significant increase produce and donations from the community to take advantage of in FBG levels (105 mg/dL, P = 8.177e-05), versus no effect on FBG for what is already available from the public to then pass on to enrich mice housed on hardwood bedding with environmental enrichment our animal’s lives. Weekly schedules are used to ensure a variety of (98 mg/dL) or hardwood bedding without enrichment (96 mg/dL). foods are provided and rotated daily to avoid repetition. There was no effect on nonfasting blood glucose levels, nor on body Adjustments are also made as needed to accommodate the specific weights of mice exposed to the different treatments. These results different research breeding groups which may require specific diets. confirm the earlier UCLA study that mice housed on corn cob Items are rigorously evaluated before being given directly to our bedding showed a significant difference in increased FBG levels animals, input comes from our service personal, both the medical versus those housed on wood chip bedding. This study also showed and behavioral, as well as the husbandry staff. While the time, effort, that enrichment made with food grade adhesives, such as those used and resources provided by the husbandry team to perform these in commercially available cardboard toilet paper rolls, showed no tasks on a continuous basis is a challenge, benefits of our enrichment effect on FBG. This information will help guide the choice of bedding program include the bonding felt by both the care staff and the

664 Abstracts of scientific papers 2019 AALAS National Meeting

monkeys. We take great pride in our dedication to create an internal website, changes are announced, and training is conducted enrichment program that utilizes many resources and provides a as necessary. There are numerous challenges throughout the process unique experience for the entire research center to enjoy. This is such as reviewer priorities, conflicting opinions, writing skills, and accomplished by providing a meaningful sense of community felt by training. The greatest challenge at our facility is completion of a everyone involved. timely review. Another training coordinator within the Animal Resources Division was brought on as cochair on the SOP committee P223 From the Technicians’ Perspective: Analyses of 2 Mouse to assist with animal specific SOPs. Different tactics have been Handling Techniques during Cage Changing attempted to improve a timely review with the most recent approach showing the greatest success. For each SOP, specific reviewers were TK Kingman*1, R Robke2, J Villano1 identified, roles were assigned, and deadlines were provided. Data of review dates demonstrates that this additional resource and focused 1Unit for Laboratory Animal Medicine, University Of Michigan, review plan has proven successful in maintaining this fluid process. Ypsilanti, MI; 2Environmental Health and Safety, University of Institutions should evaluate their current SOP program to determine Michigan, Ann Arbor, MI if the scope and design of their program is effective. It is critical to have a clear plan for the SOP lifecycle and involve personnel that are An animal husbandry technician typically changes 200 mouse cages vested in the success of the SOP and the program. daily, lifting approximately 2,080 lbs. This routine procedure and the associated animal handling equate to repetitive motions that can be P225 See Spot Run: Enriching an Aging Sled Dog Colony mentally and physically draining while also potentially causing stress on the animals. Our organization transitioned in April 2018 TL Totman*1, GA Jackson1, H Huson2 from requiring forceps use for mouse transfers to allowing technicians to directly handle mice using gloved hands to enhance 1Center for Animal Resources and Education, Cornell University, animal welfare. We performed analyses of the 2 techniques using a Ithaca, NY; 2Department of Animal Science, Cornell University, survey sent out to our husbandry technicians to evaluate preferences Ithaca, NY and several performance indices: ease of use (1=easy, 2=neutral, 3=difficult), ability to perform health checks (1=easy, 2=neutral, According to the United States Department of Agriculture (USDA) 3=difficult), and perception of being a source of cross-contamination Animal Welfare Regulations, research facilities must develop, (1=true, 2=neutral, 3=false). Factors such as participant gender and document, and follow an appropriate plan to provide dogs with years of experience were also requested. A response rate of 79% opportunity for exercise. Here, a large colony of 100 randomly (79/100 technicians) was obtained with a majority being females sourced sled dogs are housed indoors for research purposes. To stay (75%) and with more than 5 years of experience (52%). Regardless, in accordance with and exceed the AWR requirements, the sled dogs gloved hands alone were preferred overall (82%) compared to are exercised in a unique manner. Multiple groups of dogs are forceps alone (15%) and both gloved hands and forceps (3%). simultaneously exercised outdoors in 3 separate fenced play yards Statistical differences were observed between gloved hands and twice daily. Many challenges were faced developing an appropriate forceps pertaining to the ease of use (1.05 ± 0.2 and 2 ± 0.7, exercise regime such as playgroup compatibility to avoid dog fights, respectively) and ability to perform health check (1.06 ± 0.3 and 2 ± staff communication, sanitation of the play yard, adverse weather 0.8, respectively). The perception of being a source of cross- conditions, and outdoor lighting. Various playgroups were adjusted contamination was not significantly different (2.44 ± 0.7 and 2.57 ± after compatibility trials, and each group took several weeks to 0.6, respectively). Additional comments were provided by 33 successfully integrate. To prevent or stop dog fights, each yard is participants, with 30 reporting gloved hands to be better for both equipped with a catch-pole and air horns, and staff carry handheld mice (animal welfare) and personnel (ergonomics), especially as one devices for communicating. The play yard is consistently cleaned by could directly feel and apply the pressure on the mouse tail during the handler while the dogs are outside to reduce the risk of pathogen handling. Mouse bite injury reports were the same (4) for the year contamination. Incorporated into our standard operating procedures before and after the transition. Lastly, the prevalence rate of murine are alternative exercise protocols for adverse weather conditions. pathogens in our vivaria based on quarterly surveillance testing has Seasonal changes such as ground conditions and daylight hours are not significantly increased since the transition. In conclusion, direct addressed to reduce injury to dogs and staff. Developing a safe and animal handling using gloved hands presents task refinement for efficient exercise program for a colony of sled dogs is essential to the personnel without compromising animal health. dogs’ wellbeing. This pack-driven playtime regime is a unique system for addressing the exercise needs of the dogs. P224 The Life and Times of Standard Operating Procedures: Every Chapter, Something New P226 Procedure Refinement for Preparing Supply Cylinders for Germfree Isolator T Fountain*1, J McMillan2, M Thompson1 TJ Duval*1, C Vowles1, L Kennedy1, K Eaton2,1 1Environmental Health and Safety, Yerkes National Primate Research Center, Atlanta, GA; 2Division of Animal Resources, Yerkes National 1Unit for Laboratory Animal Medicine, University of Michigan, Primate Research Center, Atlanta, GA Brooklyn, MI; 2Microbiology/Immunology, University of Michigan, Ann Arbor, MI Standard operating procedures (SOPs) are born out of the necessity for a clear, concise set of procedures to follow. SOPs are fluid In our germfree mouse facility, the main method of sterilization for procedures that are continually evolving and authority for managing supplies going into germfree flexible film isolators is using a slotted SOPs as well as the scope of subjects vary between institutions. At metal cylinder with an 18-in diameter opening. The standard process our animal research facility, SOP categories range from for sealing a full cylinder for autoclaving takes precise training, administrative, safety, animal care and management, and disaster requires expensive materials, and is physically challenging for the planning. The training and compliance coordinator for the technicians. We identified a potential method of refining the process Environmental Health and Safety Office manages the SOP program to use fewer materials, be less ergonomically challenging, and most for the facility. To assure that SOPs remain current, an approach for importantly, maintain sterility of supplies entering the germfree SOP creation, review, implementation and training has been isolators. The refined procedure makes use of silicone rubber bands developed. The continual process improvement through SOP reviews to minimize the amount of high-temperature tape required, as well as is critical to the success of the program. It involves subject matter the amount of movement and twisting required in the arms and experts, a formal SOP committee, the director, and other entities such wrists. We used a clean existing isolator with sentinel mice to verify as the IACUC. Upon final approval, SOPs are published on the that the changes in procedures for cylinder prep did not compromise

665 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 sterility in the germfree mice. Once the supplies were transferred P229 Microisolator Top Bacterial Contamination Varies by Rodent from cylinder to isolator, we took weekly fecal samples for Species Yet Remains Consistent across Multiple Housing Systems microbiological monitoring (including aerobic/anaerobic cultures, gram stains, and 16s rRNA bacterial PCR). All of the results showed M Esvelt, L Steiner, CJ Childs-Thor, R Dysko, J Villano, ZT Freeman* that the isolator maintained its sterility. The refined procedure reduces the cost of wrapping a cylinder by about $190 per year just in Unit for Laboratory Animal Medicine, University of Michigan, Ann high-temperature tape. Along with the cost savings in tape, the Arbor, MI amount of time it takes to wrap a cylinder was also reduced by about 20 min per cylinder depending on the technician wrapping it which The Guide recommends sanitizing cage components, including also reduces the cost as well as the ergonomic wear on the staff. microisolator (MI) tops, at a minimum of every 2 wk. Previously published data demonstrated that mouse MI top microbial loads do P227 Procedures for Shipping Germfree Mice from an Iso-positive not increase until at least 2 wk and sanitation frequency may be Caging System delayed. It remains unclear how microbial loads differ on mouse versus rat MI tops, as well as across different ventilation systems. We TJ Duval*1, C Vowles1, L Kennedy1, K Eaton2,1, J Becker1, S Poe1 hypothesized MI top microbial loads were higher in tops from rats compared to mice and would be altered by IVC ventilation system. 1Unit for Laboratory Animal Medicine, University of Michigan, We evaluated bacterial loads on MI tops at serial time points up to 90 Brooklyn, MI; 2Microbiology/Immunology, University of Michigan, d on rat and mouse cages from static cages and multiple different Ann Arbor, MI ventilation systems (n=6 for mice, n=4 for rats). MI tops were determined to have sufficiently elevated bacterial loads to necessitate Shipping and receiving germfree mice between institutions requires changing based on either statistically significant changes in bacterial the use of a specialized sterile germfree shipping container which loads or values greater than 50 CFU. Rat MI tops across all acts as a miniature flexible-film isolator without forced ventilation. ventilation systems had significantly elevated bacterial counts The shipping isolator contains a 12-inch sleeve whereas our isolators compared to mouse MI tops at 14 days (P = 2.207e-06). Across the come standard with an 18-inch port. Transferring cages from 2 ventilation systems examined, rat MI top CFUs remained similarly different port sizes can be challenging and require more time, sterile elevated from 14 d through 90 d. Mouse MI top total CFU were also supplies, and increased risk. In order to streamline this process, we stable across multiple ventilation systems yet remained lower than 50 are piloting the use of our iso-positive caging system along with a CFU until at least 90 d. Bacterial species isolated on rat MI tops class A2 biosafety cabinet to package and ship germfree mice. The patterns were relatively consistent over time and ventilation system mice that are being shipped are transferred from standard isolators to while in mice there was greater variability in both contexts. We found iso-positive cages no more than a week before ship date. The that 14 d is an appropriate sanitization timepoint for rat MI tops germfree shipping container is ethylene oxide (ETO) sterilized in a while mouse MI tops can be extended to at least 90 d. Our data blue surgical wrap material. On the ship date, the germfree shipping highlights species differences in MI top accumulation and that mouse container and mice are placed in the hood aseptically. A technician in MI top sanitation intervals can be extended across a multitude of sterile PPE then moves the mice from the iso-positive cage into the housing systems. sterile cages used in the germfree shipping container. The shipping container is then sealed up using a standard procedure, placed in a P230 Isolator Mouse Production Facility Features and Operations shipping crate, and sent to the receiving institution. Creating a for the Exclusion of Opportunistic Pathogens process for packing mice into shipping isolators straight from iso- positive cages has helped us reduce time, materials, and R Abellas*, R Veras, R Escano, Y Hiciano, W Poueymirou, B contamination risk for mice coming out of large breeding isolators or Zambrowicz, E Levee, SE Woods mice that have been rederived into iso-positive cages without ever entering an isolator. VelociGene, Regeneron Pharmaceuticals, Tarrytown, NY

P228 Animal Identification System in a Complex Transgenic Mouse In an effort to address challenges in mouse pathogen exclusion and Breeding Colony vivarium space use, our institution recently renovated a building to include a fully autonomous, 15,000 ft2, 104 isolator mouse production R Larsen*, V Wright, K Ronellenfitch, A Nelson, M Desierto, C facility (MPF) to supply our other vivariums with opportunistic Halterman, A Keyser, T Ochoa, L Esposito pathogen-free stocks and strains. Excluding all pathogens is crucial to the integrity of our scientific pursuits, and animal health and Allen Institute, Seattle, WA wellbeing. Facility features and operations were designed and implemented to a meticulous standard of biosecurity necessary to A robust mouse identification (ID) system whose implementation exclude opportunistic pathogens. The ultimate barrier is the begins early in postnatal development enables early genotyping of semirigid, flexible-film isolators maintained under continuous transgenic mice, thereby enabling the colony manager to wean only positive pressure. Our MPF is also equipped with a variety of mice of the desired genotype(s). For this purpose, the Transgenic methods to ensure all materials entering the vivarium are pathogen- Colony Management (TCM) team has developed a system that free, if not sterile, including a clean steam autoclave and a uniquely combines paw tattooing around postnatal (P) day 10, to distinguish engineered vaporized hydrogen peroxide corridor. Only a necessary littermates prior to genotype assignment, and ear notching at number of females and vasectomized males were initially introduced weaning (around postnatal day 21). The combination of paw tattoos into the facility to be used in the surgical implantation of and ear notches provide permanent, unique identification of cryopreserved embryos. Ongoing mouse production efforts rely experimental mice. For a typical new litter, shortly after birth, each solely on internal embryo transfer and vasectomy surgeries in a pup is entered into the electronic colony management system, and specially designed operating room, colony management practices to given a unique six-digit animal ID. All pups that require genotype prevent genetic drift and any need to introduce outside mice, and analysis are then tattooed by well-trained personnel at P5-12 and at operations to package and transport mice under strict asepsis. the same time, a small tail biopsy is taken for genotype analysis. Specialty features unique to our MPF compared to traditional, less Typically, the TCM team receives all genotyping results prior to expensive vivariums enhance operations and improve risk weaning, at which point pups are ear notched with the last two mitigation; electrochromic window glass has positively impacted numbers of their unique animal ID and weaned into separate cages. staff morale as reported through feedback and our primary power This system allows for optimal usage of animal cage space in a fuel cell technology supplies dependable clean energy with a smaller complex transgenic mouse breeding colony, in part by enabling carbon footprint to sustain the building functions, with standard desirable social housing of mice from different litters without having power sources serving as back-ups. A comprehensive animal health to worry about similarities in ear notches and tattoo. 666 Abstracts of scientific papers 2019 AALAS National Meeting

and environmental surveillance program confirms our required impact of this change on microbiota, fecal and oral samples were microbial status, and a team of vivarium operations, veterinary collected before diet change and 2.5 wk after change from a services, transgenic technologies, in vitro fertilization, safety and representative subset of beagles (n=42) and subjected to 16S rRNA facilities personnel manage all known risks to our mission. The amplicon sequencing to characterize microbial community structure. features and operations of our new MPF successfully enable Body weight, body condition, and need for dietary modification were dependable, in-house sourcing of opportunistic pathogen-free mice assessed monthly for the entire colony (n=209-251). Our data show necessary for our institution’s business-critical research and heterogeneity in fecal and oral microbiota prior to diet change. development. Alteration of diet did not produce a significant impact on richness, α-diversity, or composition of fecal or oral microbial communities at P230b Glass Half Full: Hydrating Mice without Water 2.5 wk post diet transition. Dogs maintained stable body weight and appropriate body condition, with small but statistically significant CR Lockworth*, AT Pesek decreases in average body weight (10.2 kg to 9.62 kg, P = 1.5 x 10-3) and body condition score (3.12 to 3.06, P = 0.02) over a 12-mo period MD Anderson Cancer Center, Houston, TX following diet change. Over the same period the percentage of the colony requiring dietary modification to maintain appropriate body Food, water, and clean cages make up the 3-legged stool of rodent condition decreased from 34.3% to 12.9%. Our data supports the husbandry, as all are essential. Recently, we were faced with a ability to alter diet in laboratory beagles without significantly significant dilemma that would undermine this balance, as our impacting fecal or oral microbiome. This is the first study to facility was faced with an unavoidable water outage that would investigate the microbiome of laboratory beagles. affect thousands of cages that are dependent on an automatic watering system. Our entire rodent drinking water supply line P232 Platelet-endothelial Associations May Impact needed to be replaced, a process that would culminate in 3-4 d in Cytomegalovirus Replication in the Salivary Gland in Mice which no water would be delivered to our rodent colony spanning 2 buildings. To manage this, we used a multiteam approach to facilitate AM Braxton*1, J Brockhurst1,4, A Chalmin1, K Najarro1, K Johnson1, B overall plan development. Operations and husbandry teams Daly1,2, S Vijay1, G Cyphers1, C Cryer1,4, S Guerrero-Martin1, Y Su3, R explored several options and made the critical choice for temporary Arav-Boger3, KA Metcalf Pate1 water delivery: gel pouches. Once decided, we worked through the logistics of preparation, coordination, personnel training and safety, 1Molecular and Comparative Pathobiology, Johns Hopkins animal health assessment, and gel deployment. Lastly, we developed University, Baltimore, MD; 2Cummings School of Veterinary a plan to communicate the timetable, and provided reassurance to Medicine, Tufts University, North Grafton, MA; 3Department of our research community. As the water outage approached, we Pediatrics, Johns Hopkins University School of Medicine, Baltimore, implemented the plan by doubling the normal number of cage MD; 4University of Pennsylvania, School of Veterinary Medicine, change-outs in advance of the outage. Drawing personnel from Philadelphia, PA nearly all sections of our department, we were then able to open and apply gel pouches into over 18,000 cages over a span of 8 h. We then Platelet decline occurs during acute cytomegalovirus (CMV) infection focused almost exclusively on deep health monitoring of all cages in humans and mice. Platelet sequestration through association with until the water outage concluded. Upon restoration of the automatic other cells, including endothelial cells, can contribute to this decline, water supply, we determined that it was necessary to assess our and the formation of platelet-endothelial associations (PEAs) can overall management of this event in order to apply lessons learned to modify the nature of the bound endothelial cell, resulting in future events of this magnitude and scope. To evaluate our process, increased or decreased vascular permeability depending on the we collected data on dehydration cases, treatments, deaths, gel disease process. Endothelial cells become infected with CMV, serving pouches applied and replaced, and water bottles used. Data was as a site for viral persistence in multiple organs, including in the analyzed based upon room, technician, and where possible, strain salivary gland, spleen, and lungs. We hypothesize that platelet immune status. Health technicians were interviewed for feedback, aggregate formation, namely PEA formation, contributes to platelet and husbandry technicians and the research community were decline post murine CMV infection, and that the formation of PEAs administered a survey. Lastly, cost analysis and human hours will facilitate viral entry into tissues. Male BALB/c mice were required was assessed. We have concluded that our process and infected with 3 X 106 plaque forming units of the Smith strain of management plan for hydrating mice during this water outage was murine CMV. Infected mice were euthanized at 3, 8, or 21 days post- successful, due most significantly to deep health checks and inoculation and compared to uninfected controls. An increase in PEA preparation, as there was minimal impact to the health of our rodent formation was confirmed in the salivary gland at all post-inoculation population. This strategy can be applied to other facilities, both large timepoints using immunohistochemistry for CD41+ platelets (P = and small, as part of a standard disaster plan. 0.01); no alternate sites of platelet sequestration, including binding to neutrophils, monocytes, and lymphocytes, were noted using flow P231 Impact of Diet Change on Fecal and Oral Microbiome in a cytometry for platelet-leukocyte aggregates. Platelet depletion did Colony of Laboratory Beagles not change CMV titer in or affect the timecourse of CMV entry into organs, as measured by qPCR. Platelet depletion prevented AW Greenstein*1, A Ericsson2, CL Medina1 replication of CMV in the salivary glands though undepleted controls demonstrated robust replication by plaque assay (P = 0.03). 1Comparative Medicine, AbbVie, Chicago, IL; 2Department of Thus, PEA formation may enhance the ability of CMV to replicate Veterinary Pathobiology, University of Missouri, Columbia, MO within the salivary gland. Further mechanistic work is needed to determine how PEA formation affects endothelial cells and virus, Changes in animal husbandry come with the risk of introducing and whether inhibition of these interactions may provide a nonexperimental variables into a well-characterized test system. In therapeutic strategy for prevention of CMV spread. dogs, alterations in microbiome are associated with multiple disease states, and diet composition has been shown to impact fecal P233 Hematologic Values in Jamaican Fruit Bats (Artibeus microbiome. We changed to a diet with increased protein, fat, and jamaicensis) and the Effects of Isoflurane Anesthesia calories with the goal of improving efficiency in our beagle colony through reductions in food waste, decreases in the amount of canned A Strumpf*1, A Malmlov1, JD Ayers2, T Schountz1, LV Kendall1,2 food required, and less staff time spent modifying rations for increased palatability. Over a period of 1 wk, dogs were 1Microbiology, Immunology, and Pathology, Colorado State incrementally transitioned from a 25% protein dog diet to a University, Fort Collins, CO; 2Laboratory Animal Resources, calorically equivalent ration of a 27% protein dog diet. To assess the Colorado State University, Fort Collins, CO

667 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

There are more than 1,300 species of bats classified into 18 distinct P235 Use of Externalized Magnetic Ports in Mice: A Refinement for families and comprising 20% of living mammal species. There is a Longterm, Repeat-dose Intravenous Bolus Injections great deal of information regarding habitats, economic benefits, diet, captive management, and even the numerous pathogens several A Evans*, A Gaitan, P Sparks, T Gleason species of bats harbor. However, despite ongoing studies both in wild and captive populations, little is known about their normal Infusion Toxicology, Charles River Laboratories, Ashland, OH physiology. The Jamaican fruit bat (Artibeus jamaicensis), suborder Yangochiroptera, is used as an animal model for several viruses, Longterm daily intravenous dosing in mice presents many including Middle East respiratory syndrome virus (MERS-CoV), challenges. Generally, the maximum number of consecutive days dengue virus, Zika virus, and Tacaribe virus. Though Jamaican fruit mice can be successfully dosed via tail vein is 14 days. Procedurally, bats have helped advance knowledge about several infectious intravenous tail injections require a restraint device, and sometimes diseases previously stated, normal physiologic parameters are warmed gauze or water baths to dilate the vessels. Complications can lacking. This study sought to establish hematologic baseline reference include necrosis of tissues surrounding the injection site, injury due intervals in an apparently healthy, captive population of Jamaican to heating methods, and stress from restraint devices. A method fruit bats. Phlebotomy was performed in 28 awake and 15 development study was conducted in preparation for 2 anesthetized bats. Awake bats were restrained by hand and developmental and reproductive toxicology studies involving male anesthetized bats induced using isoflurane gas. Blood was collected and female fertility. The studies were to receive daily doses for a via the propetagial (cephalic) vein using a 26 gauge, ¾ inch minimum of 8 weeks (males) and between 3 and 5 weeks (females). intradermal needle. Heparinized samples were submitted for Tail vein dosing was not an option due to study length, so jugular appropriate diagnostic testing and statistical analysis performed catheters attached to externalized ports with magnetic protective using repeated measures 1-way ANOVA and posthoc Bonferroni’s caps were evaluated. The protective caps allow for breeding or social multiple comparison test with a significance level of 0.05. Results housing without damaging the ports. Twenty-five male CD1 mice were compared from physically restrained to isoflurane-anesthetized were implanted with jugular catheters attached to an externalized bats. The findings did not indicate significant differences among any port. Male mice only were used since there was no expected complete blood count parameters when comparing methods of difference in dosing administration between males and females and collection. However, the findings offer additional health monitoring the male portion of the study would be the longest. The mice were information for Jamaican fruit bats, which subsequently enhances the bolus dosed 0.2 mL of 0.9% saline approximately 5 d per wk for 13 overall understanding of their normal physiology. wk. The ports were not locked with an anticoagulant between doses. After 9 wk, 84% of the externalized ports were still patent for dosing. P234 Subsynovial Connective Tissue Development in the Rabbit At wk 13, 52% remained patent for dosing. Complications included Carpal Tunnel patency loss (2 animals), catheter dislodgement (7 animals) and skin erosion around the port (3 animals). Surgically implanted catheters AM Vrieze*1, V Schrier1,2, P Amadio2 and externalized ports allow for more consecutive daily IV bolus injections. In addition, this system would be beneficial when dosing 1Orthopedic Surgery, Mayo Clinic, Racine, MN; 2Department of irritating materials since the circulating blood volume is greater than Plastic, Erasmus Medical Center, Rotterdam, Netherlands in a peripheral vessel.

The carpal tunnel contains flexor tendons and a median nerve P236 Evaluation of 2 Surgical Models of Renal Failure in the embedded in a unique network of subsynovial connective tissue. Mouse: 3/4 and 5/6 Nephrectomy Changes of the subsynovial connective tissue’s multilayer organizational structure have been observed in both carpal tunnel A Lapierre* syndrome patients and animal research models, but little is known regarding the development of this tissue. It is unknown whether the Surgical Services, The Jackson Laboratory, Bar Harbor, ME tissue structure is an adaptation to use or if it is present in its complete form at birth and then altered from attrition over time. An established surgical model of renal failure in the mouse is the 5/6 Better understanding the development of the carpal tunnel’s nephrectomy. To reduce morbidity and mortality this model requires subsynovial connective tissue could aide in recognizing the a mouse to undergo 2 surgical procedures, a unilateral nephrectomy pathophysiology of carpal tunnel syndrome, its limitations, and the with a subsequent 2/3 nephrectomy of the contralateral kidney. potential benefit of therapeutic approaches for treatment. The rabbit Typically the two procedures are separated by a one-week recovery model helped elucidate the development of the subsynovial period. A potential refinement to this renal failure model is a 3/4 connective tissue over time. Rabbits are a common carpal tunnel reduction in renal mass conducted as a single surgical procedure. The syndrome and congenital malformation research model due to use of a 3/4 nephrectomy mouse model has been reported in the anatomical and developmental similarities to humans therefore, 7 literature as well tolerated however direct comparison of renal fetal (27-d gestation), 6 neonate (3 d), 6 juvenile (6 wk) and 6 adult function with the 5/6 nephrectomy model is lacking. The objective of (18-25 mo) rabbits were enrolled from IACUC-approved studies and this study was to evaluate these 2 surgical models of kidney failure. their carpal tunnel’s subsynovial connective tissue subjectively Five groups (n=12) of C57BL/6J male, mice at 4 and 8 wk postsurgery evaluated at macroscopic, microscopic, and ultramicroscopic levels. were evaluated. The groups consisted of group A: 3/4 nephrectomy The subsynovial connective tissue in the carpal tunnel undergoes surgery in one setting; groups B: 5/6 nephrectomy surgery in 2 marked morphological changes, from a dense, thick matrix around procedures (nephrectomy followed by partial nephrectomy); group and in between the superficial flexor tendons to a complex C: 5/6 nephrectomy surgery in 2 procedures (partial nephrectomy microorganization of interlinked sheets comprised of delicate followed by nephrectomy); group D: sham surgery in 2 procedures; flattened collagenous fibrils. The developmental patterns support the and group E: no surgery. Mice were anesthetized with isoflurane gas hypothesis that the subsynovial connective tissue structure is anesthesia and analgesics were administered (carprofen adaptive of nature and responds to loading and motion over time. subcutaneously and bupivacaine topically). Mice undergoing 2 Therefore, carpal tunnel patients might be addressed with physical surgical procedures had a 1-wk recovery period between the measures; similar to the way stress fractures can be avoided by procedures. For the partial nephrectomy procedure both poles of the avoidance of sudden increases in repetitive loading of bone. kidney were excised leaving a 4 mm remnant for the 3/4 model and a 3 mm remnant for the 5/6 nephrectomy. Body weight, kidney weight, blood, and urine samples were collected preoperatively and at 4 and 8 wk postoperatively. Data from this study demonstrate that mice in the 3/4 nephrectomy group had lower blood urea nitrogen and serum creatinine with a higher urine creatinine and kidney to

668 Abstracts of scientific papers 2019 AALAS National Meeting

body weight ratio than mice in the 5/6 nephrectomy groups. These swabs of 2 healthy kittens. We hypothesized the cats were colonized results are consistent with a more severe compromise to renal with hemolytic, genotoxin-encoding E. coli. From the colony, 27 E. function in the 5/6 nephrectomy surgical model of renal failure. coli isolates were cultured from 20 fresh feces representing the majority of cats with and without fertility failures. Samples were P237 Lysosomal Storage Disease Caused by a collected from litter boxes. Also, E. coli was cultured from a vaginal Spontaneous Hexb Gene Mutation in Immunodeficient NOD.Cg- swab. Of the 28 isolates, 21 (75%) demonstrated hemolysis on blood Prkdcscid Il2rgtm1Wjl/SzJ Mice agar. PCR was performed to determine the presence of pks, cnf, and cdt genes; 12 of the 28 isolates (42.9%) were pks+, 13 (46.4%) Z Ochoa-Dragos1, DM Imai2, L Reinholdt3, A Schile*1 were cnf+, and 10 (35.7%) were cdt+. Most isolates encoded multiple cytotoxin genes. 10 (35.7%) isolates were pks-/cnf+/cdt-, 9 (32.1%) 1The Jackson Laboratory, Sacramento, CA; 2University of California were pks+/cnf-/cdt+, 2 (7.1%) were pks+/cnf+/cdt-, and 1 (3.6%) Davis, Davis, CA; 3The Jackson Laboratory, Bar Harbor, ME was pks+/cnf+/cdt+. Whole genome sequences obtained for 3 representative isolates (2 pks-/cnf+/cdt-, 1 pks+/cnf+/cdt+) confirmed Spontaneous mutations provide a source of novel disease models in the presence of the hemolysin-associated cnf operon, the cdt operon, colonies of mice and other laboratory species. We recently identified and pks gene island. Interestingly, the pks+/cnf+/cdt+ isolate had 189 a heritable tremoring phenotype in a colony of immunodeficient base pair deletion in the cnf gene that may truncate the protein. NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice. Pedigree analysis Serine protease autotransporter toxins were also present in the showed that the condition was inherited as a recessive trait with a genomes. Antibiotic resistance genes were not identified. The median head tremor onset of 4.6 mo in both males (n=4) and females genomes were most similar to E. coli cultured from human patients (n=6). Histologic changes were consistent with a neurovisceral with urinary tract infections and bacteremia. The results from this storage disease with cytoplasmic vacuolation in neurons from the study indicate cats are colonized with pathogenic E. coli strains that central and peripheral nervous system. Cytoplasmic vacuolation was may adversely affect reproductive health and impair breeding also seen in a wider range of epithelia (gall bladder, bile duct, urinary success. bladder, renal tubular, bronchiolar, and mammary gland epithelium) P239 A Temporal Effect on Survivability in a Rodent Sepsis Model and in phagocytes within the spleen and uterine endometrium. Cryoelectron microscopy revealed additional ultrastructural changes A Cancellaro*1, T Ozment2, G Hanley1 that included membranous cytoplasmic bodies, zebra bodies, and myelin edema. Whole genome sequencing identified a 6 nucleotide 1Division of Laboratory Animal Resources, East Tennessee State deletion in the hexosaminidase B (Hexb) gene in 2 affected mice that University, Johnson City, TN; 2Department of Surgery, East Tennessee was not present in an unaffected NSG mouse or the inbred NOD/ State University, Johnson City, TN ShiLtJ reference genome. Hexb encodes a protein, beta- hexosaminidase B, that catalyzes lysosomal degradation of Sepsis has a mortality rate of 28.6%, resulting in about 215,000 sphingolipids and other macromolecules; recessive mutations of the American deaths per year and costing nearly $24 billion. Whether human homolog HEXB cause Sandhoff disease, a lysosomal storage opioid pain relief should be administered in sepsis is controversial. disease. The putative mutation, Hexblysd, was predicted to be in-frame Several publications report that opioids suppress cytokine expression and result in deletion of 2 amino acids from the protein. Genetic in rodents, thereby compromising research data. To determine if evidence for the causative role of Hexblysd was found when this opioid administration improved the welfare of septic mice without mutation failed to complement a null allele (Hexbtm1Rlp): all compromising data, we used a cecal ligation and puncture (CLP) compound heterozygous (Hexblysd/tm1Rlp) and homozygous mutant model in male C57BL/6 mice. After a ventral midline incision, the mice (Hexblysd/lysd and Hexbtm1Rlp/tm1Rlp) developed tremors, while no cecum was exteriorized and a ligature was placed one quarter tremors were observed in either heterozygous genotype during an distance from the distal end. A single 25-gauge puncture was made 8+-mo period. Median time to initial symptoms was 3.8 mo and a small bleb of cecal contents was extruded. The cecum was in Hexblysd/lysd, 4.0 mo in Hexbtm1Rlp/tm1Rlp, and 4.3 months in Hexblysd/ replaced and the incision was closed in 2 layers. Each surgery lasted tm1Rlp. Hexblysd was backcrossed 5 generations to C57BL/6J to create a 5-7 min at a rate of 10 surgeries/h. Mice were administered saline, congenic strain with similar disease incidence (median of 3.8 mo). buprenorphine HCl, or buprenorphine sustained-release polymer Our efforts describe a novel mutant mouse model of Sandhoff following surgery. Survival and indices of animal wellbeing were disease and highlight the value of whole-genome sequencing in monitored for 14 d. All groups lost a comparable amount of weight identifying spontaneous mutations. for the first few days, followed by a trend toward weight gain for the remainder of the observation period. Minimal variations in body P238 Detection and Characterization of Genotoxin- temperature were seen, with the exception of mild hypothermia in encoding Escherichia coli Isolated from Specific-Pathogen Free the saline group during the immediate postoperative period. Cats with Impaired Fertility Spontaneous movement and body posture were markedly decreased in the saline group for the first few days while both opioid treatments A Mannion*1, W McGee2, Y Feng1, Z Shen1, E Buckley-Jordan1, JG abrogated this decrease. Body condition scoring revealed no Fox1 significant differences. Opioids appeared to have a significant effect on survivability with 80% of the buprenorphine HCl mice surviving 1DCM, Massachusetts Institute of Technology, Cambridge, for 14 d while saline and buprenorphine sustained-release polymer MA; 2Laboratory Animal Resources, Colorado State University, groups had only 40% survival. Further investigation revealed a Colorado, MA significant temporal effect on survivability. It was determined that irrelevant of the treatment regimen, mortality was greatest in the While Escherichia coli are gut commensals, strains encoding virulence group that underwent surgery last. Possible explanations include factors can cause intestinal and extraintestinal disease in susceptible increased glove and instrument contamination or surgeon fatigue. In hosts. The cyclomodulin genotoxins colibactin (pks), hemolysin- summary, the use of opioids in a rodent sepsis model did improve associated cytotoxic necrotizing factor (cnf), and cytolethal distending immediate postoperative wellbeing. In this report the survivability of toxin (cdt) are associated with intestinal inflammation and cancer, the mice was more dependent on the surgical timing than whether or urinary tract infection, and septicemia in laboratory animals. The not opioid analgesics were administered. prevalence of genotoxin-encoding E. coli in cats is unknown. Over a 2-y period, an inbred laboratory colony of specific-pathogen free cats (~25) presented with resorptions, stillbirths, and pyometras in >50% of pregnancies. Hemolytic E. coli were cultured from preputial samples of 2 intact males (no clinical signs), placenta and fetal tissues of a dam with reproductive disorders, and vaginal and preputial

669 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P240 Female Urine-induced Male Mice Ultrasonic Vocalizations in showed no significant attenuation of thermal hypersensitivity. The C57BL/6J Mice as a Proxy Indicator for Postoperative Pain data suggests that both gabapentin and carprofen, given orally via flavored tablet, effectively attenuate postoperative mechanical BJ Smith*1, KE Bruner1, AM Hess2, LV Kendall1 hypersensitivity for 3 days postsurgery in a rat model of incisional pain. 1Laboratory Animal Resources, Colorado State University MIP, Fort Collins, CO; 2Department of Statistics, Colorado State University, P242 Impact of Pregnant Mare Serum Gonadotropin Fort Collins, CO Administration on Kindling Rates and Litter Size in Transgenic New Zealand White Rabbits Objectively recognizing postoperative pain in mice is challenging. Male mice produce courtship ultrasonic vocalizations in response to BL Ruelle*, S L’Italien female urine termed female urine-induced male mice ultrasonic vocalizations (FiUSV). Previous studies have shown mice do not Rabbit Operations, LFB-USA, Framingham, MA produce FiUSV while experiencing acute systemic inflammation. To determine if FiUSV can be used as a proxy indicator for postoperative Availability of commercially approved Pregnant Mare Serum pain recognition, FiUSV produced by male C57BL/6J mice were Gonadotropin (PMSG) is limited and the use of research grade evaluated once daily for 5 d before and after vasectomy or sham product in a controlled setting presents challenges. Therefore, this surgery ± sustained-release buprenorphine. Activity, orbital study was aimed at investigating the potential for removing the use tightness, posture, and piloerection were assessed at each of PMSG in a well-established synchronized artificial insemination postoperative time point. After baseline collections, 25 of 38 male (AI) protocol used in a production rabbit operation. Historically, use mice produced FiUSV 4 of the 5 d (143 ± 93 syllables, mean ± SD) and of PMSG was aimed to promote superovulation and to increase the underwent vasectomy or sham surgery. Compared to baseline (212 ± number of offspring produced. In this study, it was hypothesized that 102 syllables), vasectomized mice without postoperative analgesia elimination of PMSG would not have a significant negative impact produced significantly fewer FiUSV (59 ± 26 syllables) 4 h on the reproductive performance of transgenic rabbits (on a New postoperatively and returned to baseline by 28 h. Vasectomized mice Zealand White background) producing a human recombinant treated with buprenorphine and sham-surgery mice had no change therapeutic protein in their milk. To assess the impact of PMSG on in FiUSV from baseline at any time point after surgery. Vasectomized reproductive performance, 3 separate studies (n=90 to n=103 per mice, regardless of receiving postoperative analgesia or not, had study) were executed using nulliparous and multiparous female decreased activity at the 4-h time point compared to baseline. There transgenic rabbits ranging in age from 17 to 181 wk and the results were no differences in behavior scores between vasectomized mice pooled for each group. All animals were housed under SPF barrier and sham-surgery mice at any postoperative time point. These results conditions with a 14L:10D light cycle. The study group (n=140) was show that a decrease in FiUSV can detect decreased animal well- administered 24IU PMSG subcutaneously approximately 50 h prior being at least 4 h after vasectomy in C57BL/6J mice. to artificial insemination, while the control group (n=143) did not receive PMSG. All animals were administered an intramuscular P241 Gabapentin and Carprofen in Flavored Tablets Effectively injection of 5µg gonadotropin releasing hormone (GnRH) Provide Postoperative Analgesia in an Incisional Pain Model in immediately following AI. Endpoint parameters evaluated included Rats (Rattus norvegicus) kindling rate, litter size, and number of live kits. The results indicated that use of a PMSG treatment did not have a significant effect on B Zude*1, K Jampachaisri2, C Pacharinsak1 kindling rate (P = 0.414). Kindling rates were almost identical in the PMSG vs. control groups at 91% and 93%, respectively. Adversely, the 1Comparative Medicine, Stanford University, Stanford, PMSG group had a significantly (P = 0.003) smaller mean litter size CA; 2Department of Mathematics, Naresuan University, Phitsanulok, (8.6 kits vs. 10.0 kits) and a significantly (P = 0.011) smaller mean of Thailand live kits born per doe (7.9 kits compared with. 9.1 kits) than the control group. Based upon these results, elimination of a PMSG Providing postoperative analgesia by oral medications versus treatment regime may have a positive effect on the overall injections in rats reduces stress from frequent handling, and is less reproductive performance of the transgenic NZW rabbits. In technically challenging for investigators. The aim of this project was summary, this study suggests that removal of PMSG is an option to to investigate whether flavored tablets with gabapentin, carprofen, or consider in a commercial rabbit breeding operation regardless of the their combination effectively attenuates postoperative mechanical availability of commercially approved PMSG. and thermal hypersensitivity in a rat model of incisional pain. Male Sprague Dawley rats (n=48) were randomly assigned to 1 of 5 P243 Complete Replication of Hepatitis C Virus in BM-MSCs treatment groups: 1) placebo tablet; 2) buprenorphine sustained Derived from the Tree Shrew (Tupaia belangeri chinensis) release at 1.2 mg/kg subcutaneously once; 3) gabapentin 90 mg/ tablet; 4) carprofen 5 mg/tablet; and 5) gabapentin 90 mg and C Lu*1, Y Feng2, X Sun1, D Kuang1, N Li1, W Wang1, P Tong1, X Xia2, J carprofen 5 mg/tablet (gabapentin/carprofen). All tablets were Dai1 introduced to rats on days -3, -2, -1, 0 (surgery), 1, and 2. Rats were anesthetized via isoflurane blowby. Aseptically a 1cm skin incision 1The Center of Primate Research, Institute of Medical Biology, The was made on the plantar surface of the left hindpaw and apposed via Chinese Academy of Medical Science and Peking Union Medical suture. Mechanical (von Frey monofilament) and thermal College, Kunming, China; 2Faculty of Life Science and Technology, (Hargreaves method) hypersensitivity was tested daily and analyzed Kunming University of Science and Technology, Kunming, China on days -1, 1, 2, and 3. The amount of tablet consumed was recorded daily. Consumed tablet postoperatively was 101-133 mg/kg for Hepatitis C virus (HCV) infection is a major cause of chronic liver gabapentin, 5.5-5.8 mg/kg for carprofen, and 86-137/1.9-3 mg/kg for disease and associated cirrhosis and hepatocellular carcinoma gabapentin/carprofen, respectively. The placebo group exhibited worldwide. At present there is no prophylactic vaccine against HCV both mechanical and thermal hypersensitivities all 3 days due to lack of in vivo and in vitro model systems. A few HCV postoperatively. The buprenorphine sustained-release group genomes replicate in Huh-7 cell line and derivatives. However, these exhibited attenuated mechanical hypersensitivity for 2 days (days 1 cells are a human hepatoma-derived cell line, which is not suitable and 3) and no significant attenuation of thermal hypersensitivity. for vaccine discovery. Therefore, development of new cell systems Both the gabapentin and carprofen groups attenuated mechanical facilitating the entire HCV life cycle is urgently needed. We aimed to hypersensitivity on all 3 postsurgical days, and decreased thermal establish a novel bone marrow mesenchymal stem cells (BM-MSCs) hypersensitivity only on d 3. The gabapentin/carprofen group derived from the tree shrew to reconstruct the HCV life cycle. A tree attenuated only mechanical hypersensitivity on d 2 and 3 and shrew (Tupaia belangeri chinensis) whole-genome analysis revealed a

670 Abstracts of scientific papers 2019 AALAS National Meeting

genetic relationship between tupaias and humans. It has been including edema, hemorrhage, neuronal necrosis, and increased documented that tree shrews are susceptible to infections with HCV paracellular permeability of the blood-brain barrier (BBB). We and the primary tupaia hepatocytes (PTHs) have been also proved to investigated the pathological alterations of the BBB in rats following be infected by HCV. Although the tree shrew supports the infection single or repeated blast exposure. The BBB is composed of and replication of HCV in vivo, the HCV viremia was intermittent endothelial cells with tight junctions, a basement membrane, and the infection rate was not high (46.7%), and thus the tree shrew pericytes, and astrocyte endfeet. BINT was induced by using an was not yet established as a stable and persistent HCV model. How open-ended blast apparatus to deliver single or triple 150 kPA to refine the HCV model became an urgent problem. We transfected shockwaves to 3-mo-old male Sprague Dawley rats. After 24 h, the CD81 and OCLN, the minimal human factors required for HCV brains were excised, flash frozen, and cryosectioned into 5μm thick uptake, and miR-122 into BM-MSCs, then used a well-established sections. Qualitative and quantitative evaluation of the BBB was HCV, produced from the J6/JFH1-Huh7.5.1 culture system, to infect accomplished using immunofluorescence microscopy and Zen blue the cells. We observed that BM-MSCs transfected CD81/OCLN or analysis software. We observed alterations in expression with CD81/OCLN/miR-122 facilitates the entire HCV life cycle, including endothelial cells, tight junction proteins, tight junction adaptor infection, replication, and infectious virus production. We also found proteins, basement membrane, and pericytes. Our preliminary that the addition of exogenous vascular endothelial growth factor findings demonstrated a 1.82-fold (P = 0.037) increase in the (VEGF) could enhance its infectivity to HCV, the HCV virus load is expression of claudin-5, a tight junction protein, in the single blast up to 105 copies/mL, HCV Core antigen could be detected in the model compared to the control. There was a 2.01-fold (P = 0.018) above cell culture from day 3 postinfection. In conclusion, we used increase in the expression of PDGFR-β, a pan-pericyte protein, in the BM-MSCs, derived from the tree shrew, to establish a HCV cell single blast model compared to the control. The triple blast model model. Based on the cell model, we will reinfuse the cells which demonstrated a 1.87-fold decrease (P = 0.028) in PDGFR-β expression transfected CD81/OCLN/miR-122 in the hopes that they colonize compared to that of the single blast. This is the first comprehensive themselves in the liver, thus establishing a stable and persistent HCV pathologic analysis of the BBB in BINT in an experimental model. model in vivo. These data will support the development of a robust and reproducible experimental model of blast-induced neurotrauma. P244 How Nesting Material Is Provided Influences Barbering in Female C57Bl/6: A Pilot Study P246 Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Nonalcoholic Steatohepatitis Resolution *1 3 1,2 C Moody , D Leroux-Petersen , PV Turner C Rivera*1, S Oldham2 1Global Animal Welfare and Training, Charles River Laboratories, 2 Senneville,, Canada; Department of Pathobiology, University of 1 2 3 AST, Astrazeneca, Clarksburg, MD; Cardiovascular, Renal and Guelph, Guelph, Canada; Charles River Research Models, Charles Metabolic Diseases, Astrazeneca, Gaithersburg, MD River Laboratories, St.Constant, Canada Clinical trials assessing therapies for the treatment of nonalcoholic Nesting material is a critical resource for group-housed laboratory steatohepatitis (NASH) involve a baseline and end of study liver mice for many reasons, including thermoregulation and performance biopsy, and assessment of improvement in disease endpoints, often of highly motivated nest building activities. However, no research reflected as a percent of each treatment arm that improved, has examined how various methods of providing nest material may worsened, or remained unchanged. Traditional preclinical rodent influence mouse behavior and welfare. We predicted that distributing studies for NASH therapies are often limited by not knowing the nest material around the cage would allow mice equal access to the level of liver disease/NASH present at the start of therapeutic material, thereby reducing aggression and improving mouse welfare, intervention, instead randomizing treatment groups on easily compared to nest material provided in one area of the cage. In this measurable endpoints such as body weight or metabolic status. We pilot study, 6-wk-old female C57Bl/6 mice (n=27; housed 3 per cage) describe a liver biopsy technique in a diet-induced NASH mouse were randomly assigned to 1 of 3 groups: 1) standard (1 tissue; n=3), model, for the assessment of baseline liver disease in order to exclude 2) nest puck (8g brown crinkle paper + one tissue; n=3), or 3) mice that do not exhibit fibrosis and to equally distribute animals dispersed (8 g brown crinkle paper sprinkled throughout cage + one with similar fibrosis between treatment groups. These levels can then tissue; n=3). Cage change occurred every 7 d over the 15-d test period be compared to terminal, postintervention levels for a truer and new nest material was provided. On the last day of testing, mice understanding of in vivo pharmacological effects and thus more were scored for presence/absence of wounding and barbering. No accurately reflect clinical trial design strategies. The mouse is wounds were detected on mice throughout the duration of the study, anesthetized (Isoflurane inhalation, 2-3% to effect) and sterilely thus this parameter was not further analyzed. Statistical analysis of prepared for surgery. A small incision is made in the upper abdomen barbering was conducted at the cage level. The results show that all and the left lateral lobe of the liver is exposed. A wedge of liver cages of mice in the nest puck treatment group showed no barbering, (approximately 10-30 ug of tissue) is surgically removed and a while all cages of mice in the other 2 treatment groups had obvious similar-sized piece of absorbable gelatin is put in its place to stop any barbering (missing patches of fur; P = 0.037). This preliminary study bleeding. The mouse is surgically sutured, analgesia is administered suggests that providing nest pucks may help protect female C57Bl/6 (0.1 mg/kg buprenorphine) stapled closed, and will recover back to mice from barbering, when compared to dispersed crinkle paper or normal (visual observations for qualitative physical activity and provision of a tissue alone. Further examination is needed to eating/drinking behavior) within 1 day. Biopsy and terminal liver elucidate the behavior of mice housed in these treatment groups, in pieces were fixed and stained via standard protocols and assessed by addition to replication of the current study results. a pathologist according to the NASH Activity Scoring (NAS) System. To showcase the value of this surgical procedure, it was used in a P245 Blood-brain Barrier Pathology in an Experimental Model of study that examined the impact of a glucagon-like peptide-1 (GLP-1) Blast-Induced Neurotrauma receptor agonist on NASH endpoints. A total of 118 mice were

* biopsied, and 49 of those were excluded due to low or high fibrosis. C Cheatham , T Lyle, G Uzunalli, A Dieterly, R Shi, M Hoang, S Herr The remainder of the mice were sorted into groups for treatment. GLP-1R agonist treatment was associated with overall improvement College of Veterinary Medicine, Purdue University, West Lafayette, of fibrosis with 17% of the treatment group improving and 83% IN remaining unchanged. Similarly, the treatment improved the overall NAS score with 66% of the group having improved NAS score and The signature injury of modern warfare is blast-induced 33% remained unchanged. Additionally, GLP-1R agonist treatment neurotrauma (BINT) due to the use of explosive devices. The clinical improved inflammation, with 75% of the group having a lower symptoms of BINT have been correlated with neuropathology inflammation score and 25% remained unchanged. We exemplify the

671 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 utility of this procedure by leveraging the pre-study biopsy to assess (median nesting temperature after injury without temperature the impact of therapeutic treatments on NASH endpoints in mice. management was 34.1°C). Median (range) brain weight was 7.8g (5.4-8.9g, n=15) in injured males and 7.0g (4.8-8.2g, n=16) in injured P247 Evaluation of Long Acting Buprenorphine for Analgesia in females, compared to 9.0g (8.0-10.1g, n=6) and 7.7g (7.0-8.1g, n=6) in Mice respective controls (P < 0.05 for both). Injury was associated with widening of the longitudinal fissure and narrowing of the coronal K Patil*1, LV Kendall2, CJ Doane1 and anterior ectosylvian gyri. Animals with injury displayed decreased speed and activity in the open field and increased lateral 1University of Arizona, Tucson, AZ; 2Colorado State University, Fort deviation from the midline during locomotion on the catwalk. This Collins, CO model of inflammation-sensitized HIH injury in the neonatal ferret provides a range of macroscopic injury, resulting in significant gait Buprenorphine is a widely used opioid analgesic for postprocedural deficits and decreased spontaneous activity in the open field. pain in laboratory mice. The standard formulation of buprenorphine Preventing hypothermia after HIH significantly contributes to injury. HCl (0.3mg/ml) is typically administered every 8 to 12 , requiring This model can provide a platform to assess therapies for brain injury repeat handling. As such, the use of compounded sustained-released in near-term infants. buprenorphine has increased to provide up to 72 h of continuous analgesia following a single injection. Although convenient, P249 HER2-positive Luminal B Metastatic Breast Cancer in a acquisition of compounded controlled substances is challenging Mouse Model especially with the emergence of state laws directed at opioid addiction. Further, compounded buprenorphine formulations have DL Donohoe*1, J Yin3, SD Konduri2 variable efficacy, strain-related variance, and have not been tested in our most common inbred strains. Buprenorphine HCl 1.8mg/ml is 1Neurooncology and Translational Research, Aurora Health Care, an FDA-approved veterinary drug labeled for 24-h analgesia in cats. Milwaukee, WI; 3Oncology, Aurora Health Care, Milwaukee, WI In order to assess the potential use of the FDA-approved formulation in mice, we compared buprenorphine 1.8mg/ml with the More than 300,000 women per year are diagnosed with breast cancer. compounded sustained-release buprenorphine to define the Risk factors include genetics, age, and hormone dependent/reactive pharmacokinetics of these drugs in C57BL/6 mice. Vendor purchased conditions. Among hormone receptor-positive breast cancer 7-wk-old C57BL/6 male mice received 1 of the following treatments subtypes, HER2-positive Luminal B patients are often associated (3 mice per group): buprenorphine 1.8mg/ml (0.9mg/kg SQ) or with the worst prognosis, primarily due to the presence of estrogen sustained-release buprenorphine 0.5mg/ml (1mg/kg SQ). Blood was receptor, progesterone receptor, and human epidermal growth factor collected for analysis of systemic buprenorphine levels at 2, 4, 8, 12, 2 (HER2). However, if treated appropriately and timely, the risk of 24, 36, 48, and 72 h post initial drug administration from 3 mice per recurrence and metastasis in patients with luminal B breast cancer time. Plasma concentrations of buprenorphine were determined by could be significantly reduced. Here, we proposed a strategic liquid chromatography mass spectrometry. Results indicate that the approach for developing animal xenografts which could be used to FDA-approved formulation had a peak concentration of 29 ng/ml at faithfully represent the response to treatment on patients and could 2 h, which dropped to 12 ng/ml at 4 h, then maintained a also be used to validate other potential treatments. We used a HER2- concentration >1ng/dl up to 12 hours. The sustained release positve luminal B patient breast tumor resected from the metastatic buprenorphine had a peak concentration of 3.1 ng/ml at 2 h, which site in brain to derive tumor spheres. Once growth of the spheres was slowly declined to less than 1 ng/ml between 12 and 24 h post confirmed, the spheres were orthotopically injected (5x105cells) in 2 administration. Our pharmacokinetic data imply that the FDA- groups of nu/nu mice (4, 6-wk-old female, 4, 3-mo old) at cerebellum approved buprenorphine may be a suitable alternative to the site, corresponding to the original tumor resection site. Animals were compounded sustained-release product for up to 12 hours before imaged monthly with MRI over a 20-wk period to monitor tumor subsequent dosing may be required. growth. Animals were euthanized if they presented with neurological symptoms, or increased tumor burden. The 2 goals of this pilot were P248 Characterization of a Novel Neonatal Ferret Model of to assess tumorigenicity of Her2-positive patient tumor in age- Prematurity matched animal models to investigate whether age plays a role in tumorigenesis of this breast cancer subtype and to generate a self- D Moralejo*1, T Wood1, K Corry1, C Fisher1, P Parikh1, J Snyder2, S propagating PDX cell line to test efficacy of 2 anti-cancer agents, Juul1 trastusamab and MK 1775 and investigate associated mechanisms. Our pilot study suggests a difference in age between animal cohorts. 1Pediatrics, University of Washington, Seattle, WA; 2Comparative Younger mice developed tumors in within 8-12 wk while older mice Medicine, University of Washington, Seattle, WA failed to develop measurable tumors. PDX cells in-vitro demonstrated more sensitivity to MK1775 than patient cells which The neonatal ferret is an attractive species in which to model preterm provides opportunity to identify mechanisms for treatment to brain injury because it has a gyified brain with a white-to-gray matter decrease metastasis in HER2+/luminal B breast cancer subtype. ratio similar to the human (unlike rodents). Postnatal white matter maturation and complex cortical folding in ferret first 4 postnatal wk P250 Femoral Venous and Arterial Catheterization in an Ovine are equivalent to that observed in the human brain during the third Fetus for Serial Blood Draws trimester. Postnatal day (P) 17 ferrets are considered comparable to 32-36 weeks’ human gestation, the age at which over 80% of preterm E Pollack*1,2, A Nedder1 infants in the U.S. are currently born. Our goal was to characterize behavioral testing paradigms and gross pathology after a severe 1Boston Children’s Hospital, Charles River Laboratories, Urbana, inflammation-sensitized hypoxic-ischemia-hyperoxic (HIH) insult in IL; 2University of Illinois College of Veterinary Medicine, Urbana, IL the P17 neonatal ferret. P17 ferret kits received 3mg/kg of LPS followed by bilateral carotid artery (CA) ligation and consecutive The goal of our study was to determine the best catheterization HIH (30 min at 9%, 30 min at 80%, 30 min at 9%) before reversal of method for serial fetal blood samples to determine plasma drug the right CA ligation. Controls received saline and normoxia. At P42, concentrations after maternal dosing. Pregnant Dorset sheep at a gait and spontaneous activity were determined in an automated gestational age range of 95-120 days were used. The study required catwalk and open field. A gross pathology score was developed to blood collection from the fetus for a minimum of 7 d. Two methods characterize cortical injury. Development of gross macroscopic injury of fetal catheterization were performed, both accessed via required 6h of temperature management at normothermia (median hysterotomy. In group 1 (n=3 fetus), the fetus was positioned and the 37.0°C, range 36.0-37.7°C) to prevent spontaneous hypothermia head and neck exteriorized allowing for visualization of the external

672 Abstracts of scientific papers 2019 AALAS National Meeting

jugular vein and carotid artery. A small cut down was performed and P252 Buprenorphine Effectively Provides Postoperative Analgesia the jugular and/or carotid were catheterized using a 4Fr sheath- in an Incisional Pain Model in Neonatal Rats (Rattus norvegicus) introducer kit. In group 2 (n=3 fetus), the fetus’s posterior limbs were located prior to making uterine incision and then the limbs were EM Katz*, M Huss, K Jampachaisri, C Pacharinsak exteriorized from the uterine horn to allow for catheterization. The femoral vessels were exposed via the cut down method and a 3Fr Comparative Medicine, Stanford University, Stanford, CA PICC was placed and secured. In both groups, an extension line was placed and tunneled through the uterus and maternal abdominal There is limited information on safe and effective neonatal rodent wall and secured via purse string. The uterine wall was closed with analgesia. The aim of this study was to evaluate the efficacy and consideration for cotyledon and caruncle using a TA uterine stapler. duration of analgesia provided by buprenorphine HCl (Bup HCl) The uterine horn was placed back in the abdominal cavity, maternal and lidocaine (LDC) in an incisional pain model. Male and female laparotomy was closed. Fetal blood was sampled every 4-12 h and postnatal d 3 Sprague Dawley rat pups (n=40) were randomly the extension line was heparin locked with 25 u/mL at a minimum of assigned to 1 of 5 treatment groups: 1) saline - 0.1 ml subcutaneous every 12 h. Femoral catheterization was highly favorable and more (SC) + 0.01 ml saline incisional infiltration; 2) BupL - 0.025 mg/kg successful with a 100% fetal viability and consistent reliable blood Bup HCl SC + 0.01 ml saline incisional infiltration; 3) BupH - 0.05 draws. The method of jugular/carotid line placement did not reliably mg/kg Bup HCl SC + 0.01 ml saline incisional infiltration; 4) LDCL allow for blood sampling over the duration of 7 days. There were a - 0.1 ml saline SC + LDC 2 mg/kg incisional infiltration; and 5) variety of reasons for this including inability to properly secure the LDCH - 0.1 ml saline SC + LDC 4 mg/kg incisional infiltration. lines, fetal rotation with entanglement, and the natural bend of the Subcutaneous doses of Bup HCl or saline were administered neck of the fetus which would impede sampling. With femoral preoperatively under anesthesia. Rats were anesthetized with catheterization, the line was secured to the leg which eliminated sevoflurane by mask, and a 10 mm full thickness skin incision was patency issues. Additionally, the length of the catheter was much made over the left lateral thigh. Incisional infiltration of LDC or greater allowing for it to be seated in the abdominal aorta/vena cava saline occurred prior to closure with surgical glue. Baseline thermal for improved sampling and overall patency of the catheter. latency was measured 24 h prior to surgery, and subsequently 1, 2, 4, 8, 24, and 48 h postoperatively using an infrared diode laser. Thermal P251 Development of an End Stage Rat Model of Retinal latency in the saline group was significantly reduced compared to Degeneration for Development of Autologous Cell Replacement- baseline through the 8-h timepoint. Pups in both LDC treatment based Therapeutics groups had similar thermal latency times as the saline group. Both 0.025 and 0.05 mg/kg Bup HCl thermal latency time was statistically E Kaalberg*, I Han, M Ulferts, L Wiley, E Burnight, RF Mullins, EM similar to baseline for the entire 48-h period. Thus, a single Stone, BA Tucker preoperative dose of Bup HCl 0.025 to 0.05 mg/kg SC effectively attenuates postoperative thermal hypersensitivity in a neonatal rat Ophthalmology, University of Iowa Institute for Vision Research, incisional pain model. North Liberty, IA P253 Using PET Imaging to Assess the Safety Profile of Inherited retinal degenerative disorders such as retinitis pigmentosa Intravesical-dosed Bladder Cancer Therapies are characterized by death of the light sensing photoreceptor cells of the outer neural retina. Like the rest of the central nervous A Alfaro*, K Kuszpit, E Straley, D Goldsteen system, the endogenous regenerative capacity of the retina is limited, and as a result, photoreceptor cell death causes debilitating Animal Sciences and Technologies, AstraZeneca, Gaithersburg, MD irreversible blindness. Although stem cell-based photoreceptor cell replacement strategies have been quite successful experimentally, Each year in the U.S. over 70,000 new cases of urothelial carcinoma when transplants are performed in hosts with advanced disease, (UC) of the bladder are diagnosed. While many treatment options donor cell survival, and in turn functional integration, is often exist, response rate and durability of response to standard therapies poor. We hypothesize that transplanting autologous cells on remains low. The recent FDA approval of 5 immune checkpoint support scaffolds will mitigate this problem. To test this hypothesis inhibitors (CPIs) targeting PD-1/L1 has created the potential to novel animal models of end stage disease, with eyes large enough significantly improve outcomes in UC patients. Given the increased to receive subretinal grafts, are required. Three rat models of and unnecessary risks involved with systemic administration of CPIs inherited retinal degenerative blindness were generated on the to UC patients with localized, nonmetastatic disease, we aimed to Sparge Dawley background via CRISPR-based deletion of the quantitatively assess how much if any intravesical (IVS)-dosed CPI genes Pde6b, Rpgr, and Rho. Animals were genotyped via genomic therapy “leaks” into systemic circulation. PET imaging is a method PCR. OCT imaging and histopathology were used to characterize that can provide a sensitive and quantitative measure of radiolabeled the rate of retinal degeneration in each strain (n=54, 3 male drug biodistribution. We radiolabeled a CPI therapeutic antibody and 3 femail at 1, 3, and 6 mo of age for each strain). Deletion with 89Zr for PET detection. To determine how much if any IVS- of Rpgrand subsequent loss of Rpgr protein did not significantly dosed drug leaked into circulation, we injected 3 groups of mice alter photoreceptor cell viability at any of the ages analyzed. While (male C57Bl/6, n=6/group) with a therapeutically relevant (3.8 mg/ the Rho knockout model was found to have significant retinal kg) dose of drug: 1) 100uL IVS-administered drug, 2) 200uL IVS- degeneration at 6 months of age, these animals had excessive weight administered drug, and as a control, 3) 150uL IV-administered drug. gain and premature death. Upon further investigation mammary IVS dosing was accomplished in anesthetized mice (2% isoflurane and pituitary tumors, indicative of CRISPR induced off-target editing mixed in oxygen) and mice were kept under anesthesia for 3 h prior during generation, were detected. The Pde6b knockout animals to and then during PET scanning. To ensure that radiolabeled IVS- had a significant retinal degeneration phenotype, with extensive injected test article did not leak outward from the bladder, the penis photoreceptor cell death detected as early as 1-month post-birth. In of each mouse was ligated immediately after dose administration. conclusion of the 3 rat models generated the Pde6B strain had the Without being allowed to wake, mice were euthanized via

most aggressive disease with almost complete loss of the retinal CO2 administration after PET scanning. In control (IV injected) mice, outer nuclear layer by 4 weeks of age. Rapid progression to end stage normal systemic biodistribution concentrated the radiolabeled drug disease is ideal for testing of novel therapeutics and will be used in in liver and spleen (63 and 32% injected dose/gram (%ID/g), downstream pre clinical experiments. respectively) with little if any signal observed elsewhere. In the low volume (100uL) IVS-dosed mice, radiolabeled drug remained exclusively in the bladder without any appreciable extra-bladder biodistribution (0.5, 0.1, 0.5 %ID/g in liver, spleen, and kidneys, respectively). In the high volume (200uL) IVS-dosed mice, there was

673 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 significant retrograde leakage via the ureters into the renal pelvis of fog and removed at multiple time points ranging from 15-min to 4-h the kidneys (~18 %ID/g), and ultimately into central circulation exposure time. A different set of tapes were exposed to an iHP where radiolabeled drug exhibited normal systemic biodistribution misting surface sprayer system with slides exposed to 1, 2, or 3 to the liver and spleen. These data support that dose volume is an treatment applications with 7 min contact time each. Only the important factor, and that at lower volumes, IVS dosed-CPIs can exposed portions of the ova were treated. After exposure, ova were remain in bladder with little if any leakage into systemic circulation incubated in a hatching medium for 6 h. Control tapes were thus ensuring an optimal safety profile. maintained at room temperature without fog/mist exposure before incubation in the hatching medium. After incubation, the number of P254 Vagotomy Increases Astrocyte Branching in the Nucleus ova hatched was assessed by microscopic examination. Exposure to Tractus Solitarii and Modifies Hypoxic Responses both environmental and surface sprayer systems using iHPe decreased the number of Syphacia spp. ova hatched vs controls. The GC Hofmann*1,2, EM Hasser3,2, DD Kline3,2 15-min, 30-min, 90-min, and 4-hr time points showed an average of 43 to 51% reduction in hatching for the room fogging 1Comparative Medicine Program, University of Missouri, Columbia, decontamination system. The surface-treatment samples averaged 36 MO; 2Area Pathobiology, University of Missouri, Columbia, to 46% reduction in hatching depending on conditions. Conversely, MO; 3Biomedical Sciences, University of Missouri, Columbia, MO only 13.5% of the eggs on the control slide did not hatch. These data suggest that exposure to iHP fog and mist is effective in reducing The nucleus tractus solitarii (nTS) in the brainstem is the initial site of hatching of Syphacia spp. ova at the time points tracked. Further integration and modulation of sensory input from the studies are needed to determine whether changes to exposure cardiorespiratory system. The nTS is important in the body’s parameters of iHP treatments can further reduce or eliminate response to hypoxia, and changes in nTS function are associated with hatching, and infectious potential, of rodent pinworm ova. diseases such as obstructive sleep apnea, hypertension, and heart failure. Vagal afferents supply visceral sensory information to the P256 Sterility and Stability of Diluted Meloxicam in Compounded nTS and activate both neurons and astrocytes to modify neuronal Multidose Vial after 365 Days activity. Vagotomy decreases nTS neuronal activity, but whether this is a direct effect on neurons or indirect due to altered astrocyte H Kawano*4, G Simonek1, AD Moffitt3, J Tahara2, L Brignolo3 structure or function is unknown. We hypothesized that decreased afferent input (vagal nerve transection) modifies nTS astrocyte 1Seattle’s Children, Seattle, WA; 2California Animal Health & Food morphology, and is associated with impaired cardiorespiratory Safety Laboratory, Davis, CA; 3Campus Veterinary Service, responses to hypoxia. Male Sprague-Dawley rats (5-7 wk) were University of California, Davis, Davis, CA; 4University of California, exposed to hypoxia while conscious (8-12% O2, 5 min each) or Davis School of Veterinary Medicine, Davis, CA anesthetized with isoflurane (90 s, 12% O2) and cardiorespiratory parameters were measured. Rats then received right-sided cervical Meloxicam is a common analgesic used for rodents. Since meloxicam vagotomy (n=7) or sham surgery (n=7). Hypoxic challenge was is only commercially formulated for companion animals, it requires repeated following 1 wk of recovery. Although respiratory responses dilution to provide appropriate doses to smaller laboratory species. A were similar in conscious animals, unilateral vagotomy blunted compounded multidose vial (cMDV) is often created to dilute and respiratory response to hypoxia under anesthesia compared to store a diluted drug. However, chronic cMDV use runs the risk of presurgery measurements. Rats were euthanized and brains were contamination and becoming a potential source of nosocomial processed for immunohistochemistry with an antibody against infection. In this study, we created 15 cMDVs by diluting meloxicam astrocytic glial fibrillary acidic protein (GFAP). GFAP with sterile water (dilution 1:10). cMDVs were punctured once daily immunoreactivity in the left (intact) and right (vagotomized or sham) for 30 d. To determine the sterility of the diluted meloxicam, 8 nTS was compared. Vagotomy increased astrocytic GFAP expression cMDVs were assessed for bacterial growth on days 0, 10, 20, 30, and in the nTS compared to the intact side and to shams (paired T-test). 365 and were tested for endotoxin on days 0, 30, and 365. The Sholl analysis was performed on nTS astrocyte tracings. Vagotomy stability of 7 cMDVs were assessed on days 0, 10, 20, 30, and 365, increased number of endpoints and overall branching at the more using liquid chromatography-diode assay. No bacterial growth or distal processes on the denervated side compared to shams, endotoxin were detected at any time point and the drug indicating a morphologic response. These data suggest decreased concentrations remained stable up to 365 d. Based on this study, we afferent input modulates nTS astrocyte morphology which may believe that cMDVs of diluted meloxicam can remain sterile and influence cardiorespiratory function. Changes in afferent input thus stable for 365 d. may play a role in cardiovascular and respiratory diseases, and modifying these changes could contribute to treatment regimens. P257 Evaluation of C-peptide and Insulin Levels as Markers of Treatment Efficacy in a Human Amylin Transgenic Mouse Model P255 Room Decontamination Using Ionized Hydrogen Peroxide of Type-2 Diabetes Fog and Mist Is Effective in Reducing Hatching of Syphacia Ova J Aitken*, T Nie, GJ Cooper G Dell’Anna*1, B Ball2, K Mullin1, M Sauer1 School of Biological Sciences, University of Auckland, Auckland, 1Laboratory Animal Resources, Iowa State University, Ames, New Zealand IA; 2Office of Animal Resources, University of Iowa, Iowa City, IA Type 2 diabetes is a problem impacting all populations but with We evaluated the efficacy of ionized hydrogen peroxide (iHP) fog increasing risk in developing countries where changes in society and mist for environmental and surface decontamination have led to changes in diet and a more sedentary lifestyle. At the of Syphacia spp. ova in rodent rooms. Ova were collected by perianal very early stages of diabetes or prediabetes, changes in diet and tape impression of Syphacia-infected mice. Tapes with attached ova exercise can have an impact on disease progression. Early were exposed to iHP fog using a whole-room fogging intervention with compounds that slow down/ameliorate diabetes decontamination system in unoccupied housing rooms of 2,600 ft3 progression and enable internal glucose homeostatic mechanisms to and 1,890 ft3, respectively. The air handling system was disabled at take control would be ideal. Rodents are not naturally diabetic, so we the room level and air intake, exhaust ducts, and doors were sealed constructed transgenic mice that overexpress the human amylin (hA) with tape and plastic sheeting. Each room received an initial fogging gene. Amylin is a hormone cosecreted with insulin from pancreatic injection time of 15 min at 0.5 ml/ft3/min of hydrogen peroxide. beta cells and along with insulin, is involved in glucose homeostasis. Iodine test papers were placed throughout the entire room to confirm The transgenic mice become spontaneously diabetic and are used as fog penetration. After injection time, ova were continually exposed to a model of type 2 diabetes to test plant-derived proprietary

674 Abstracts of scientific papers 2019 AALAS National Meeting

compounds for their ability to slow diabetes progression. As there are determine whether the root cause of the event can be directly several compounds to test in vivo, our aim was to develop a clinical attributed to the investigational product, or if there are other biomarker which would enable determination of initial efficacy, contributing factors. We postulated that other possible factors leading before progression to a long-term preclinical study. The compound to unexpected animal mortality/adverse events may include was administered in drinking water ad libitum to hA transgenic underlying congenital disease, subclinical infections (acquired prior (n=13) and non-transgenic (n=9) mice from age 21 d. Water-fed to receipt), experimental parameters leading to stress and stress transgenic (n=13) and nontransgenic (n=9) littermates were used as related pathology, or equipment, technical, and sanitation issues. To controls. Weight, blood glucose, fluid intake, and core body help determine the cause of these events, our facility performs temperature were recorded and serum was collected from tail bleeds diagnostic necropsies and laboratory testing on animals found dead at 50 and 100 d of age. C-peptide and insulin ELISA assays were or euthanized in extremis. To evaluate the effectiveness of our performed on serum samples. C-peptide concentration was a better process, we retrospectively data mined diagnostic cases from the past biomarker than insulin for identifying treatment effect. C-peptide 5 y (2014-2019), including gross and microscopic findings, levels increased significantly with time (50 to 100 d) in water-fed signalment, clinical observations, and duration of time since arrival transgenic mice (P = 0.028) but not in compound-treated transgenic at the facility and study initiation. A definitive diagnosis was mice (P = 0.20). A combination of both C-peptide and insulin levels established for approximately half of the cases (n=124), while the over time gave the best comparison of treatment (C-peptide x remaining half had no lesions or nonspecific findings. Many of the insulin: P = 0.008 compared with P = 0.056; water versus treated undiagnosed cases had a clinical sign of neurologic symptoms but transgenic mice). High sensitivity ELISA assays require small the diagnostic necropsy failed to identify a lesion, which might be a amounts of serum which are collected from tail bleeds. C-peptide result that we only collect the brain as our standard CNS tissue and insulin assays were used to determine the initial efficacy of a collection for our diagnostic necropsy. We believe that by examining compound in vivo, reducing the time needed for testing and are a additional neural tissues, such as spinal cord and peripheral nerves, refinement of long-term survival assays. we will increase the chances of identifying lesions and thus decrease the rate of nondiagnostic outcomes for these cases. Reducing the P258 Cell Line Xenograft Growth in the B6;129- number of cases with undiagnosed mortality, will result in more Rag2tm1FwaIL2rgtm1Rsky/DwlHsd (R2G2) Mouse Model meaningful data, with less repeat work required and allow for definitive interpretation of the safety of new agents. J Naden* P260 Dosing Fenbendazole in Water for the Treatment of Mouse Veterinary Science, Research and Support, Envigo RMS Inc, Pinworms (Syphacia obvelata and Aspiculuris tetraptera) Indianapolis, IN J Duggan*1,2, KR Yachera1, CE Roache1, MC Terzi1,2, T Coksaygan1,2, Immunodeficient mouse models are helping to advance the field of LJ DeTolla1,2, NL Schmidt3 oncology. A new model on the market, the B6;129- Rag2tm1FwaIL2rgtm1Rsky/DwlHsd (R2G2) knockout mouse, lacks 1Comparative Medicine , University of Maryland, Ellicot City, responsiveness to common gamma chain cytokines, including IL-2, MD; 2Pathology, University of Maryland, Baltimore, MD; 3School of IL-4, IL-7, IL-9 and IL-15. In addition, this model exhibits defects in Medicine, University of Maryland, Baltimore, Baltimore, MD lymphoid development and so lacks mature lymphocytes of the B, T, and natural killer (NK) cell lineages. Herein we describe growth and Syphacia obvelata and Aspiculuris tetraptera are common take rate of a few tumor cell line xenografts in the R2G2 nonpathogenic pinworms of laboratory mice. Although infestations immunodeficient mouse model. The cell line xenografts studied are classically subclinical, heavy burdens can lead to rectal prolapse, include multiple myeloma, pancreatic, Burkitt’s lymphoma, ovarian, intussuception, and enteritis. Additionally, a potential research and breast cancer. Multiple myeloma (MM1.S cells) were inject into 5 complication is immunosuppression which leads to unnecessary male mice via the tail vein at 50 x 104 cells per mouse. IVIS imaging variability or discrepancies in data. Fenbendazole is a was used to measure tumor burden of luciferase transfected MM1.S methylcarbamate benzimidazole antiparasitic agent that has been cells. Imaging was completed weekly until the tumors reach 1000 used to treat murine pinworms through premedicated feed. mm3. The pancreatic cancer (BxPC-3 cells) was injected Traditionally, pinworm infested mice are treated with 150 ppm of a subcutaneously into 5 male mice at 1 x 106 and tumor volume was fenbendazole-impregnated diet for 5 wk. However, premedicated measured via calipers weekly. Burkitt’s lymphoma (RAJI) cells, feed is costly and therefore unfeasible for most institutions to ovarian (A2780) cancer cells, and breast (T47D, MDA-MB-231, and implement. Because of this, our goal was to determine an effective MCF-7) cancer cells were implanted into subcutaneously at 1 x dose of fenbendazole for use in mouse drinking water. To achieve 106 cells in both flanks of 5 female mice each. Tumor volume and this, we tested a fenbendazole oral suspension that is approved for body weight was measured twice weekly via calipers for 5 w. use in the drinking water of poultry. During the study, mice were Average tumor growth reached the 1000 mm3 threshold by day 20-35 kept under strict quarantine conditions. These conditions were postinjection for all cell lines tested. These data provide evidence that established by only exposing mice to their home cage on an the R2G2 mouse model is a valuable tool for oncology programs individually ventilated rack or a biosafety cabinet in the animal including cell line tumor models research, with high take rates and facility. Naturally infected mice with positive PCR results were quick growth of allogeneic models. confirmed to have an active infestation by perineal tape tests. We then prepared 3 different doses in water pouches (150 ppm, 300 ppm, P259 Investigating Central Nervous System Collections as a and 450 ppm). Ten mixed-gender C57BL/6J mice were used for each Contributing Factor to Unsolved Diagnostic Necropsy Cases dosing condition. For each cage, we measured water consumption weekly. We resuspended water pouches weekly and observed daily JL Reel*3, J Justen2, M Magagna1 for suspension particles. After 2 wk of PO dosing, we performed perineal tape tests and fecal flotations from the ceca and colons of the 1Department of Pathology, Charles River Laboratories, Mattawan, infested mice. At this time, all mice tested negative for pinworm Mattawan, MI; 2Experimental Therapeutics, Charles River eggs. No changes were observed in water consumption and no Laboratories, Mattawan, Mattawan, MI; 3Post-Life Services, Charles suspension particles were noted in the water pouches. Based on our River Laboratories, Mattawan, Mattawan, MI results, we recommend treatment using fenbendazole in mouse drinking water and strict quarantine conditions for an effective The FDA requires nonclinical safety assessment testing in animal elimination of pinworm infestations. models to occur before new drug products or medical devices can proceed into phase 1 human clinical trials. Unanticipated mortality and adverse effects in these studies warrants investigation to

675 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P261 Effects of Sedative and Anesthetic agents (Dexmedetomidine, gavage: the control group, administered with 200 μl of distilled Alfaxalone, Propofol) on Urodynamic and Anesthetic Parameters water/polyethylene glycol, and the genistein group, administered in Male Cats with 200 μl of distilled water/polyethylene glycol containing 150 mg genistein/kg/day. The first dose was given 7 d following the JJ Xu*1,2, Z Yousuf2,3, Z Ouyang2,3, T Martin1, PA Lester1, TM Bruns2,3 xenografts, when tumors were palpable. In each week, the dosage was administered for 5 consecutive d, followed by 2 d of rest. At the 1Unit for Laboratory Animal Medicine, University of Michigan, Ann end of treatment, animals were euthanized and blood was collected Arbor, MI; 2Biointerfaces Institute, University of Michigan, Ann by cardiac puncture. Using histology and immunohistochemical Arbor, MI; 3Biomedical Engineering Department, University of analyses as well as EIA technique for hormonal determiantions, the Michigan, Ann Arbor, MI antitumoral effects of genistein were determined for 3 wk on an inflammatory mammary cancer xenograft model. No statistically Urodynamic studies in animals are essential to understanding the significant differences in tumor size were found between control and pathophysiology of bladder disorders. Cats are commonly used to treated mice. Distant metastases were significantly higher in study urinary disorders and treatments due to their physiological genistein xenografts versus control group. Significant differences in similarity with the human system. However, sedative and anesthetic Ki-67 index were found between treated and control xenografts being agents used for these studies are known to influence bladder higher in genistein group. Serum progesterone, androstenedione, and function. We compared the effects of commonly used anesthetic estrone sulphate levels were also in the genistein group. agents on urodynamic and anesthetic parameters. Five adult male Intratumoural and serum estrogens were elevated, whereas cats were sedated 3 times with each of 3 agents: dexmedetomidine intratumoural testosterone levels were decreased. These results (0.02–0.04 mg/kg intramuscular [IM] bolus; reversed with matched revealed that genistein ingestion promotes tumour proliferation and volume of atipamazole after session), alfaxalone (5 mg/kg IM bolus elevates metastatic rates by increasing intratumoural and circulating + 0.08 mg/kg/min intravenous [IV] constant rate infusion [CRI]), estrogen levels in a mammary cancer xenograft model. and propofol (2 mg/kg IV bolus + 0.15–0.2 mg/kg/min IV CRI). Two cystometrograms (2 mL/min infusion of body-temperature saline) P263 Using Epstein-Barr Virus DNA to Predict Lymphomagenesis were conducted in each sedated session, and urodynamic parameters in PDX Models (peak pressure, bladder capacity, bladder compliance, pressure trend) and anesthetic parameters (heart rate, respiratory rate, blood J Li, SH Lee* pressure, time to lateral recumbency, time to head up, and time to ambulation) were evaluated. No significant differences were NARLabs, Tainan, Taiwan observed between peak pressure, bladder capacity, or compliance. Qualitatively, bladder pressure increased gradually before voiding Patient-derived xenograft (PDX) model is recognized as an accurate with propofol and alfaxalone, and spiked just before voiding under and clinically relevant animal model for precision medicine. The PDX dexmedetomidine. Trials under alfaxalone had a greater number of model is the only system that can accommodate full heterogeneity nonvoiding contractions. Each anesthetic agent provided adequate and variability of solid tumors. Therefore, it is often used to simulate chemical restraint, and anesthetic parameters were within normal tumor growth in human. The PDX model has been hindered by limits. Dexmedetomidine had the shortest time to lateral recumbency, spontaneous lymphomagenesis, which is highly suspected to be as well as the shortest time to head up and ambulation after reversal caused by EBV. Due to the formation of lymphomas in PDX model, or discontinuation of the agent. However, cats were more likely to precision cancer treatment and drug development could vomit after administration of dexmedetomidine. In summary, all consequently lead in a false direction. We found that 38 of 178 (21%) agents adequately sedated cats for cystometrograms, and gave PDX tumors were EBV-associated human lymphoma by IHC similar peak pressure, bladder capacity, and bladder compliance (Immunohistochemistry) of antihuman CD20. In addition, we found values. Qualitative differences were seen in the pressure trends, and a strong correlation between EBV DNA copy number and PDX differences in time to induction and recovery were observed between lymphoma formations. The quantity of EBV DNA can be a predictor agents. These subtle differences may be a reason to choose 1 for PDX failure. In order to validate the hypothesis, a total of 25 adult anesthetic agent over others, depending on the nature of the study. mice were used for xenografts, and the samples from PDX mice included plasma and tumor tissues. When the tumor volume reached P262 Changes in Steroid Hormone Profile and Tumor Progression 500 ~700mm3 in PDX mice, blood was obtained from the facial vein after Genistein Treatment of Canine Inflammatory Mammary by using a lancet in an EDTA-coated tube. Plasma was separated by a Cancer Xenotransplanted into Mice centrifuge for a qPCR analysis. The mice were euthanized when the tumor reached ~1000 mm3 and the tumor tissues were collected for A Alonso-Diez*1, A Martin-Ruiz1, G Silvan1, M Illera1, L Peña2, A lymphomagenesis IHC analysis. From the qPCR of EBV DNA of Gonzalez-Gil1, I Diez-Prieto3, C Perez-Garcia3, S Caceres1, JC Illera1 tumor tissue and plasma samples, we found that samples collected from tumor tissue have a higher predicative accuracy comparing to 1Animal Fisiology. Fac Veterinaria, University Complutense of samples from plasma. However, the samples collected from plasma Madrid, Madrid, Spain; 2Medicina y Cirugia ANimal, Universidad can be useful as an early indicator for lymphomagenesis. In Complutense de Madrid, Madrid, Spain; 3Dpto. Medicina, Cirugía y conclusion, although the link between EBV and lymphoma is clear, Anatomía Veterinaria, Universidad de León, Leon, Spain the accuracy of diagnosis and prevention of lymphoma transformation in PDX models remains to be revealed. Isoflavones, such as genistein, have been proposed to provide beneficial effects on health, including preventive or therapeutic P264 Transponder Implantation on Postnatal Day 1 Rat Pups actions in carcinogenesis. Their structural similarity to estrogens allows them to bind at the cellular level with estrogen receptors. This J Muehleisen*, J Brum, J Smith study attempts to determine the antitumoural effects of genistein for 3 wk in a canine inflammatory mammary cancer xenograft model, in Envigo, East Millstone, NJ terms of tumor proliferation, appearance of metastases, and steroid hormones regulation. Twenty-four nonovariectomized female SCID Rat pups are typically identified by ink injections in their paws on mice (BALB/cJHan®Hsd-Prkdcscid), 6-8 wk old and weighing postnatal day 1 (PND 1). However, the ink can fade making between 20 and 22 g were used. The xenograft was directly identification difficult and the paws can become permanently established from a 9-y-old female dog with a spontaneous damaged as a result of the technique. This trial evaluated a new inflammatory mammary carcinoma. Tumor fragments were procedure to identify pups by implanting IMI-500 radio frequency subcutaneously implanted into the ventral side of 3 female SCID identification (RFID) transponders (microchips). The microchips were mice. Xenografts from third passage were treated for 3 wk by oral placed subcutaneously with a 15-gauge needle in the scapular region

676 Abstracts of scientific papers 2019 AALAS National Meeting

and surgical glue was used to aid in closing the injection site. A nonsterile approach was taken for the animals as well as the The Cre-Lox recombination system has been used to generate tissue- microchips. This technique was performed on 3 different litters for a specific and time-dependent mouse strains for gene knockout studies total of 33 Sprague Dawley pups. Clinical observations were done on and remains an important tool for many researchers. These models the pups on days 1-7, 14, 21, and 70. Body weights were taken on rely on ligand-dependent Cre recombinases that are activated by days 1, 4, 7, 14, 21, and 70. Five/33 pups had a scab and/or swelling administration of tamoxifen. Tamoxifen is not benign however; it is at the injection site and needed to be implanted again as the original hepatotoxic and problematic to solubilize for injectable delivery microchip fell out. The pups had no abnormal signs by time they which is the preferred route of administration. While alternatives to were weaned on d 21 and all microchips were still intact and injectable delivery exist that are theoretically less stressful, such as working. Body weights were normal and no other notable administration through the feed, researchers cite weight loss and observations were made. Surface righting reflex was done on d 2 and inconsistent induction in their mouse strains when feed the pups were handled by the scruff as they would be for oral gavage administration is used. We, therefore, sought to understand what dosing on d 7. The microchip did not affect these procedures. The change in weight might occur following administration of injection technique was refined throughout the trial to determine commercially available tamoxifen-containing feeds in C57BL/6J how far to insert the needle to ensure the microchip stayed in but not mice. Male and female mice of 2 age groups were placed on either too far to injure the pup. In summary, transponder implantation is an 400 mg/kg tamoxifen citrate diet (TAMC, n=19) or 500 mg/kg effective method of identification for PND 1 pups and will minimize tamoxifen diet (TAM, n=21). The “young” subset of these mice were identification issues, allow for direct data capture, and eliminate the 7 wk of age (n=6 for each diet) and the “old” subset ranged between risk of trauma to the pups’ limbs and digits. 20 and 24 wk of age (n=13 on TAMC and n=15 on TAM). All mice P265 Anesthesia Effects on Prairie Voles (Microtus ochrogaster) experienced significant and sustained weight loss during the 7-d period after placement on these diets. Average weight loss exceeded K Rapp-Santos*1, V Chaplin1, M Normann2, G Pratt1, M McWaters2, 20% of baseline weight by d 6 on both diets in young mice and WT Watanasriyakul2, O Akinbo2, A Grippo2, N McNeal1 exceeded 20% by d 5 in the adult group given TAM. Greater average weight loss was observed on TAMC compared with TAM diet (P = 1Naval Medical Research Unit San Antonio, San Antonio, 0.04). A linear mixed model analysis was considered for weight loss, TX; 2Department of Psychology, Northern Illinois University, DeKalb, with main effects for diet, age, sex, and time. The potential for IL 2-factor interactions was also considered. The analysis revealed significant main effects for diet (P =0.01) and days (P = <0.01), and Prairie voles (Microtus ochrogaster) represent a potential translational significant interactions between diet and days (P = 0.03) and days model for examining the physiologic and behavioral effects of and age (P = <0.01). We can conclude from this study that significant various stimuli as they engage in several social behaviors that mirror weight loss does occur when mice of this strain are placed on 400 those of humans (e.g. living in family groups, cooperative rearing of mg/kg tamoxifen citrate diet or 500 mg/kg tamoxifen diet. More offspring, development of long-term pair bonding, mate-guarding) investigation on the efficacy and side effects of lower dose diets and and regulate their autonomic nervous system in a manner similar to alternate routes of administration should be undertaken to identify humans. Specifically, prairie voles may be useful in examining how the most humane way to induce Cre recombinases in these models. different social stressors may affect response to acute trauma and recovery from injury. For example, disruption of social bonds has P267 Estradiol Enhances the Inhibitory Effect of the Cytokine been shown to induce cardiovascular dysfunction in this species, Interleukin-1β on Pulsatile LH Secretion in Female Mice similar to humans. While some limited hematologic information exists for some species of voles, normal blood gas values and K Makowski*1, R McCosh2, M Kreisman2, K Breen2 chemistries have not been published for the prairie vole, especially in response to inhalational anesthesia. This study measures the impact 1Animal Care Program, University of California, San Diego, of prolonged inhalation anesthesia on heart rate, blood gases and Escondido, CA; 2University of California, San Diego, San Diego, CA chemistries, and behavior of the prairie vole. All subjects (n=30) were housed in male/female pairs. Baseline paired animals (n=18) were Inflammatory and infectious processes can disrupt reproduction briefly anesthetized with isoflurane followed by cardiac puncture for function and fertility in a wide range of species. Immune cells blood collection. Post anesthesia pairs (n=12) were anesthetized for produce cytokines in response to systemic stressors and these 2.5 h, followed by a 20-min recovery from anesthesia post-observed cytokines have the potential to disrupt the central mechanisms righting reflex. Arterial whole blood was collected and processed controlling gonadotropin secretion and ovulatory cyclicity in females. using CG4+ and Chem8+ cartridges on the i-STAT handheld blood When administered peripherally, interleukin 1 beta (IL-1β) has been analyzer at baseline and end of anesthesia for comparison. Significant shown to inhibit the proestrus luteinizing hormone (LH) surge in differences were observed in several values when comparing baseline intact female rats. However, in male gonadectomized rats, IL-1β and post-anesthesia samples. Lactate decreased from 7.7 ± 3.0 delivered via intraperitoneal (IP) injection does not alter LH mmol/L to 4.9 ± 2.3 mmol/L, BUN increased from 26 ± 2 mg/dL to secretion, so the effect of IL-1β may depend on the presence of 38 ± 8 mg/dL, and glucose decreased from 194 ± 53 mg/dL to 128 ± gonadal steroids. Here, we began by investigating whether 31 mg/dL. Additional findings indicated that anesthesia significantly peripheral IL-1β can inhibit mean LH secretion in female mice and if decreased heart rate over time and impaired performance in the estradiol is necessary for this inhibition. Female C57bl6 mice (10 wk) open-field behavioral test when comparing pre- and post-test effects. were ovariectomized and implanted with a silastic capsule This study generated baseline arterial blood gas and chemistry containing oil (n=12) or 100ng estradiol (n=9), which approximates a values in healthy prairie voles that will be critical in examining the diestrus level. LH was measured prior to and at hourly intervals effects of traumatic injury and other research models, and also following IP administration of 20ng/g IL-1β (n=11) or saline explored the influence of anesthetic drugs on prairie vole blood (control,n=10) from tail-tip blood samples. In oil-treated females, parameters. mean LH did not significantly change following saline or IL-1β, compared to pretreatment levels. In contrast, IL-1β elicited a P266 Weight Loss in C57BL/6 Mice Fed Tamoxifen and Tamoxifen significant reduction in LH at 120 and 180 min following Citrate-containing Diets administration. These data indicate that estradiol is necessary for peripheral IL-1β to suppress LH. Based on these findings, we K Patil*1, E Bedrick3, E Torabzadeh2 conducted two experiments. First, we performed a study (n=13) to measure pulsatile LH prior to and following IP administration of 1University Animal Care , University of Arizona, Tucson, AZ; 2BIO5 20ng/g IL-1β or saline (control) in ovariectomized C57bl6 mice Institute, University of Arizona, Tucson, AZ; 3Epidemiology and pretreated with estradiol. We saw a significant decrease in mean LH Biostatistics, University of Arizona, Tucson, AZ and LH pulse frequency in mice pretreated with estradiol. Second,

677 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 we investigated the neural pathways whereby IL-1β may access the in a Broome restrainer in lateral recumbency. The tail was taped onto kisspeptin and GnRH circuit to influence LH pulsatility using c-Fos a mouse restraint platform, and a winged 27x1.5in catheter was taped as a marker. We saw a significant increase in c-Fos in the in place. The animal was anesthetized and moved to the MRI scanner paraventricular nucleus (PVN), middle arcuate (mARC), where images were acquired using a 3D-TWIST sequence, and 0.25ml ventrolateral medulla (VLM), and nucleus tractus solitarii (NTS). of Gadoteridol was administered as a contrast agent via tail vein These data suggest that estradiol affects the specific neural pathways catheter. Ambient air temperature, animal temperature prior, and by which stress hormones influence reproduction. post sequence was then taken. Image series were reconstructed into 4D series and time series regions of interest extracted for tracer P268 A Comparison of the Efficacy and Cardiopulmonary Effects of kinetic modeling to determine regional blood flow. Based on our 3 Different Sedation Protocols in Otolemur garnettii initial findings, mice with thermal jackets retain greater body temperature, and thus higher regional blood flow, when compared to KR Finnie*2, CP Jones2, WD Dupont1, KJ Salleng2, KA Shuster2 the nonjacketed group. This suggests body temperature should not only be reported, but also controlled, in all studies where brain 1Department of Biostatistics, Vanderbilt University, Nashville, perfusion is used for tracer or medication distribution to reach TN; 2Division of Comparative Medicine, Vanderbilt University therapeutic levels. Medical Center, Nashville, TN P270 Tns2 Is a Tumor Suppressor Gene that Regulates Intestinal The Northern Greater Galago (Otolemur garnettii) is a prosimian Tumor Progression in Apc Deficient Mice primate studied to better understand the evolutionary development of vision and somatosensation. Since little has been published about K Hiura*1, A Sakanoue1, S Kontani1, Y Takahashi1, K Nakano2, T physiologic parameters under sedation/anesthesia in this species, the Okamura2, S Kakuta3, H Sasaki1, N Sasaki1 goal of this project was to compare 3 sedation protocols commonly used in other nonhuman primate species. Specifically, this study 1Kitasato University, Towada, Japan; 2National Center for Global aimed to investigate the efficacy and cardiopulmonary effects of Health and Medicine, Shinjuku, Japan; 3The University of Tokyo, alfaxalone (Alf; 8 mg/kg IM) compared to ketamine (Ket; 20 mg/kg Tokyo, Japan IM) alone or ketamine + dexmedetomidine (Ket/Dex; 4 mg/kg + 25 μg/kg IM) with reversal (atipamezole; 250 µg/kg IM). A total of 34 TENSIN (TNS) is a family of multidomain scaffold protein that bind animals were evaluated, including 11 juveniles and 23 adults. Indirect the cytoplasmic tail of β-integrins, and localize to adhesions that blood pressure, heart rate, respiratory rate, SpO2, and rectal anchor stress fibers in cells. In TNS family, TNS2 is ubiquitously temperature were measured. Blood was collected for blood gas expressed in human and mice. We conducted data mining using the analysis and a blood chemistry panel. Induction time, immobilization COSMIC, the exsome database of human tumor. Among various time, and recovery time were recorded. Subjective measures of tumor cases, we detected a considerable number of cases quality and efficacy were evaluated including quality of induction, of TNS2 gene mutation like nonsynonymous mutation or nonsense pedal withdrawal reflex, palpebral reflex, muscle tension, and quality mutation in the mining of the intestine tumor cases. To evaluate of recovery. All 3 protocols successfully immobilized the animals and whether Tns2 is tumor suppressive, we bred Apc deficient all animals recovered from sedation. Heart rates were highest among and Tns2 deficient mice to create combined mutation mice, and the Ket group and the lowest within the Ket/Dex group. The Alf examined the number of polyps they develop and the level of group had the longest immobilization time which was, on average, malignancy of these polyps. Number and size of intestinal tumors twice the time of Ket or Ket/Dex. The Ket/Dex group had an average were significantly increased in compound mutant mice. Tns2 is a recovery time one-third of the other groups and subjectively had the multidomain protein composed of a PTPase (PTP) domain at the best quality of recovery. Based on these results, we recommend Ket/ N-terminus, and Src homology 2 (SH2)/phosphotyrosine binding Dex over Alf or Ket alone for brief sedation of healthy galagos. As (PTB) domains at the C-terminus. However, little is known about the with any sedation/anesthesia protocol, care should be taken to function of both domains. Thus, we examined the function of each choose the best drug combination for each individual animal. domain in tumor suppression using TNS2-deficient HCT116 colon tumor cells. We introduced Tns2 wild-type (WT), PTP inactivated P269 Thermal Body Regulation and Its Effects on Regional Blood mutant (C231S), SH2-PTB domain deleted mutant (ΔC) to HCT116. Flow Perfusion Using Dynamic Contrast Enhanced Magnetic Significant reduction of migration was observed in WT and C231S. Resonance Imaging On the other hand, a significant decrease in the proliferation was observed in WT, but no difference was observed in C231S and ΔC. K Eldridge*, R Speedy, AA Bedwell, P Territo These data suggest that each domain of Tns2 has the distinct function for a tumor suppressor. In conclusion, Tns2 is a tumor suppressor Radiology, Indiana University School of Medicine, Indianapolis, IN gene in this in vivo mouse model and likely has a similar role in human colon cancer. Imaging of brain perfusion using Dynamic Contrast Enhanced Magnetic Resonance Imaging (DCE-MRI) has been used for nearly 30 P271 Evaluating the Infection Model of Chlamydia trachomatis in y in preclinical studies. In animal models, anesthesia can result in C3H/HeJ Mice changes in core body temperature, which is known to alter regional brain perfusion. In an effort to assess the impact of core body KJ Riebe*, BM Ricci, U Gautam, GD Sempowski temperature on regional perfusion 20 C57BL/6J mice were randomly allocated to 1 of 2 groups. In the first group 5 males and 5 females Duke Human Vaccine Institute, Duke University Medical Center, were imaged without heat loss control. The second group of 5 males Durham, NC and 5 females were jacketed in a custom MRI compatible thermal cover to minimize thermal loss. Observations indicated that mice Chlamydia trachomatis (Ct) is a bacterial pathogen that causes upper without thermal jackets lost on average 5-6C (females=6C; males=5C) genital tract of infection leading to oviduct inflammation and over 30 min. Temperature was taken via external temperature probe subsequent infertility in women. Currently there is a critical need for every 10, 20, and 30 min. By contrast, mice with thermal jackets the development of novel antibiotics and vaccines to combat the retained overall greater body heat, losing only 2-4 degrees continued rise in the rate of STDs, such as Chlamydia. To that end (males=2C; females=4C) over the same interval. In order to quantify there is an urgent need to characterize and standardize a mouse regional blood flow in male and female C57BL/6J mice, DCE-MRI model of Ct infection, a human pathogen that does not typically images were acquired on a scanner outfitted with a 4 channel phased infect wild-type laboratory mouse strains. Using C3H/HeJ mice array head coil. Prior to being placed onto the MRI scanner (endotoxin resistant, 6-wk-old) we investigated whether intravaginal vasodilation was obtained via heat lamp and the animal was placed infection with Ct would induce a humanlike infection in the upper

678 Abstracts of scientific papers 2019 AALAS National Meeting

genital tract and cause lesions and could therefore serve as a more investigated. We compare the effect on the microbiome of using robust model of Ct pathobiology for vaccine and therapeutic topical triple antibiotic ointment (TAO) or hypochlorous acid gel development. In this study, 6 mice were randomly assigned in 2 (0.01%) on our UD cases. Twelve UD cases were randomly assigned groups. The groups of 3 mice each were either Ct infected or to 2 groups, 6 mice treated with TAO and 6 mice treated with uninfected controls that receives buffer (SPG). In this model the hypochlorous acid gel. The mice were colony animals on C57BL6 bacterial colonization and replication was measured in vaginal swab backgrounds belonging to multiple investigators. Skin swabs were shortly after the intravaginal delivery of 20 µL of 1 million inclusion collected aseptically by swabbing the hair and skin of the right flank forming units (IFUs) of Ct, using a permissive HeLa cell-based assay. from each mouse with a sterile swan. Swabs were placed in DNAse/ Histopathology results demonstrated hydrosalpinx, characteristic of RNAse free tubes containing 1 mL of ultrapure water. Fecal pellets Ct infection. Clearance of infection was observed in all Ct challenged- were collected directly from the rectum and placed into empty tubes mice by day 25. Together we show that C3H/HeJ mice induce using sterile forceps. All samples were placed on dry ice immediately hydrosalpinx, disruption of epithelial architecture, and recruitment after collection and then stored at -80 C until sequencing. Baseline of neutrophils in genital organs, suggesting intravaginal infection as samples were taken before therapy was started. Control swabs were natural route of C. trachomatis infection and replication. Use of this also collected to rule out environmental bacterial contaminates model and defined phenotype will substantially accelerate discovery due to collection technique. Mice were treated once daily with the of next generation antibiotics and vaccines to fight a growing epi- respective medication until resolution of lesions. Two weeks after demic. the topical therapy was stopped, posttreatment skin swabs and fecal pellets were collected. DNA extraction, preparation of 16S rRNA V4 P272 Correlating Nesting Behavior and Body Temperature region amplicon libraries, and sequencing were performed. Analysis to Behavioral Assessments for Endpoint Determination of the sequence data showed that mice treated with TAO had a in Burkholderia Pseudomallei-infected Mice statistically significant reduction in bacterial counts and diversity on their post-treatment skin swabs and fecal pellets. Mice treated with K Jenner, KE Bruner, LV Kendall* the hypochlorous gel showed minimal impact. Comparison of the 2 medications for efficacy of treatment showed no statistical difference. Laboratory Animal Resources, Colorado State, Fort Collins, CO Our study provides strong evidence that hypochlorous gel is a good treatment choice for UD when microbiome sparing is desired. Mice are frequently used as an animal model to evaluate preventative and therapeutic compounds against Burkholderia, a high-priority P274 Assessment of Experimentally Induced Sinusitis in Rabbits bacterial pathogen. Developing methods to assess humane endpoints Using a Novel CT Scoring System in these studies can be challenging. Ethograms are frequently used to assess the welfare of mice and include observations such MA Hull*1, S Cole1, AK Brice2, U Sajjan3, R Duran-Struuck1,2 as general appearance and activity, facial expressions, rearing, grooming, and nesting activity. Body temperature can also be a 1Pathobiology, University of Pennsylvania, Glenolden, PA; 2ULAR, useful tool to assess mouse wellbeing during infectious disease University of Pennsylvania, Philadelphia, PA; 3Temple University, studies. To evaluate the correlation between nest building, body Philadelphia, PA temperature, and other ethogram assessment, mice used in a dose escalation study of Burkholdeira pseudomallei were observed prior Over 38 million U.S. adults were diagnosed with rhinosinusitis in to and after inoculation. Five mice in each group were inoculated 2016, a condition that impacts the quality of life for sufferers, and with either 5,000, 10,000, or 2,5000 CFU. Mice were observed for 3 with indirect costs of decreased productivity in excess of $12.8 d prior to inoculation, then at 4, 12, 20, 28, 36,45, 53, and 61 h after billion dollars per year in the U.S.in 2016. Advanced imaging inoculation for the behaviors listed above, nesting activity, and body and symptomatic scoring have improved accurate diagnosis and temperature. All mice, regardless of dose, had a sharp decline in treatment in man, but this methodology has not been demonstrated general appearance, activity, rearing, grooming, and nesting activity in the most used translational animal model, the rabbit. We beginning at 12 h postinoculation. Orbital tightening increased developed a novel scoring system of CT image analysis to quantify during the first 12 h post inoculation. Body temperature began to the severity of experimentally induced rhinosinusitis and confirmed drop between 20-28 h postinoculation. By 28 h postinoculation, such severity via histopathology. CT scans were taken of New appearance, activity, grooming, and nesting activity were absent. Zealand White rabbits with 0.5 mm sections. Nine animals were Orbital tightening scores increased after inoculation. Body imaged on day 0 and 7 days following a surgical infection of the temperature reached it nadir at 36 h and did not increase through maxillary sinus with S. aureus. Animals were either treated with hour 61. The decline in nesting activity correlated well with the saline or a novel nitric oxide based experimental compound. Images appearance (r=0.87), activity (r=0.83), rearing (r=0.69) and body were scored using a Zinreich modified Lund-Mackay (ZLM) scoring temperature (r=0.75). There was also a negative correlation to orbital system on 3 key areas, the maxillary sinus, middle meatus, and tightening (r=-0.23). Similarly body temperature correlated well with nasopharyngeal duct. Preliminary data shows that animals infected appearance (r=0.8), activity (r=0.85), rearing (r=0.69), and a negative with bacteria had a significant increase in radiopacity within sinus correlation to orbital tightening (r=-0.22). These data demonstrate structures when compared to sterile controls, with a 10-fold change that nest building activity and body temperature can be used as a in Hounsfield units. Similar to humans, such results correlated with proxy indicator for animal wellbeing in studies with Burkholderia to an increased ZLM score, the current gold standard of diagnosing establish humane endpoints. sinusitis. To determine if CT scores correlated with gross and histopathological examination following treatment with a novel P273 Comparison of Topical Treatments for Ulcerative Dermatitis experimental compound, tissue was harvested on day 14 (after 7 d and Their Effect on the Microbiome in C57BL6 Mice of treatment) at the time of euthanasia. The tissue was fixed in 10% formaldehyde, decalcified, sectioned, and stained with H&E, gram L Colenda*1, P Subramanian2, K Rice1, JM Smith1, JM Izzi3, H stain, and periodic acid-Schiff to evaluate tissue structure, bacterial Habbershon1, CG Linton1 burden, and mucin production. Preliminary histopathological analysis using Necrosis, Epithelial Degeneration, Inflammation, 1DVR, NIH/OD/ORS, Bethesda, MD; 2OCICB, National Institutes Osteonecrosis (NEDIO) scoring demonstrated that treated animals of Allergy and Infectious Diseases, Bethesda, MD; 3Research Animal had >50% improvement of their sinusitis compared to saline controls. Resources, Johns Hopkins University, Baltimore, MD In summary, we have refined the rabbit preclinical sinusitis model by using a human CT scoring system and preliminary studies support Ulcerative dermatitis (UD) is a common condition in C57BL6 research that NO treatment improves sinusitis which makes it attractive as a mice. The rise of antibiotic resistance due to overuse of antibiotics nonantibiotic based therapy. is a growing concern and nonantibiotic treatments for UD are being

679 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P275 Neurochemical Characterization of the Spontaneous Mutation (T) readings acquired by conventional IT and IC. The IR devices Tremor, a New Mouse Model for Epilepsy were used to measure T in 25 obese mice (OM), averaging 54 g, and normal weight (NM) mice, averaging 26.2 g, from the proprietary MS Garcia Gomes1, PK Yamamoto1, JM Alemán Laporte*1, DA strain. Unexpectedly, OM showed T averaging 1.23°C higher than Zanatto1, TM Sandini2, JC Florio2, I Lebrun3, SG Massironi4, SR NM using the IT, while the IC data showed a 0.68°C lower T than Alexandre-Ribeiro4, M Bernardi5, CM Mori1 NM (P < 0.01). Both IR devices presented consistent T readings with a low variability. In addition, the IT showed higher T readings 1Department of Pathology, School of Veterinary Medicine and than IC by an average of 1.2°C (P < 0.01). Even though we had a Animal Science, University of São Paulo (USP), São Paulo, , significant difference in T readings between the devices, we cannot Brazil; 2School of Pharmaceutical Sciences, University of São Paulo conclude differences are solely due to the adipose layer. We believe (USP), São Paulo, Brazil; 3Biochemistry and Biophysics Laboratory, the higher T finding in IT could be due to the heat generated by the Scientific Development Division, Butantan Institute, São Paulo, digestion of the high fat/high sugar diet the OM are fed, or by the Brazil; 4Department of Immunology, Institute of Biomedical Science, larger amount of heat producing brown fat the mouse is known University of São Paulo (USP), São Paulo, Brazil; 5Graduate Program to form. We conclude that although IR devices can be a reliable in Environmental and Experimental Pathology, Universidade welfare oriented technology to evaluate BT in mice, an internal Paulista, São Paulo, Brazil standardization should be used to show intrinsic deviations for location, strai,n and type of research. More experiments are needed The tremor mutant phenotype results from a recessive spontaneous to further understand the changes and to establish a standard mutation that arose from a colony of Swiss mice. The mutant method to reliably use IR devices to measure BT. mice did not differ from WT mice in physical and reflexology development from birth to 3 wk of age. Motor impairment P277 Characterization of D. musculi Burden and Serum IgE was reported in adult mutant along with tremor, ataxia, and Concentration in BALB/cAnNCrl Mice Pre- and Post-treatment spontaneous seizures that can be observed around postnatal day with Imidacloprid/Moxidectin 21 and worsen with aging, evincing a neurodegenerative pattern of this mutation. Considering characteristics analyzed so far, M Gugel*1, RJ Ricart Arbona2,1, W Boteler3, CL Perkins4, KS this mutation might represent an animal model for spontaneous Henderson4, N Lipman2,1 seizures. So to quantify neurotransmitters expressed in this animal, and to create a better understanding of this mutation 1Tri-Institutional Training Program in Laboratory Animal Medicine development and evolution, has been made necessary. The genome and Science, Memorial Sloan-Ketting Cancer Center, Weill Cornell scan using microsatellite markers distributed through the mouse Medicine, The Rockefeller University, New York, NY; 2Memorial genome showed that the tremor mutation is on chromosome 14, Sloan Kettering Cancer Center, Weill Cornell Medicine, New York, in the interval of 5 cM between D14Mit37 (33.21) and D14Mit115 NY; 3Immuno Probe, Frederick, MD; 4Charles River Laboratories, (38.21 cM), making Egr3 (Early Growth Response 3) the main Wilmington, MA candidate gene. To quantify neurotransmitters in hippocampus, 8 animals were euthanized by decapitation and the contents of Sharing genetically engineered mouse strains has led to the noradrenaline (NOR), vanillylmandelic acid (VMA), dopamine (DA), inadvertent introduction of Demodex musculi into many colonies. 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), Detection methods were previously unreliable and its presence in 5-hydroxytryptamine (5-HT), and 5-hydroxyindole acetic acid (5- laboratory mice went unrecognized. While infestations are clinically HIAA), and the amino acids GABA, glutamate, glycine, and aspartate inapparent in most mouse strains, significant serum IgE elevation has were determined using high-performance liquid chromatography been shown in Swiss Webster, C57BL/6NCrl, and BALB/cAnNCrl (HPLC). Regarding the dopaminergic system, neither the DA nor mice, with the most profound elevation in BALB/cAnNCrl. Topical DOPAC content, and the turnover rate of DA (the ratio of DOPAC application of imidacloprid/moxidectin at 13/3mg/kg is an effective plus HVA to DA), showed a statistical difference between WT and treatment for D. musculi in TRP1/TCR mice after 8 consecutive mutant mice. The concentration of NOR was significantly increased weekly treatments. We sought to determine if this treatment (P = 0.0012) and the ratio VMA/NOR was decreased (P = 0.032) eliminated mites in BALB/cAnNCrl mice and would return IgE in the hippocampus in the mutant compared to WT mice. The concentration to preinfested levels. BALB/cAnNCrl mice were concentration of 5-HT (P = 0.0083) and 5HIAA (P = 0.0032) in mutant housed with a Demodex-positive NSG mouse for 42 d, after which mice was significantly increased in the hippocampus compared with animals were treated with topical administration of imidacloprid/ that WT mice. The contents of GABA (P = 0.0123), glutamate (P = moxidectin or vehicle (vehicle controls) to the interscapular region. 0.0217) and aspartate (P = 0.0124) were significantly increased in Another group of uninfested animals were treated (treatment the hippocampus in the mutant compared to WT mice. The glycine controls). Animals were confirmed Demodex-negative prior to the levels were decreased in the hippocampus (P = 0.0168) of mutant experiment via PCR. Swabs for PCR, serum for IgE concentration compared with that WT mice. In conclusion, the increased levels determination, and pelt biopsies for mite burden assessment were of neurotransmitters GABA, glutamate, aspartate, NOR, 5-HT and collected 7 and 28 d following cessation of treatment (n=5/group/ 5-HIAA in the hippocampus suggested that an impairment of Egr3 timepoint). Treated infested animals did not have evidence of gene function could explain the mechanism involved in seizures and mite infestation via PCR or histology at either timepoint and 80% tremor observed in the mutant mouse, a potential model to study of vehicle controls were negative by day 28. IgE levels of infested epilepsy. animals were markedly elevated and decreased significantly 28 d after cessation of treatment, but remained approximately 3-fold P276 Obesity in Mice Affects Temperature Readings When Using 2 higher than in uninfested animals. IgE levels remained unchanged Types of Infrared Thermometers in treatment and vehicle controls. Though treatment is effective in clearing mite infestation, serum IgE levels likely require longer J Hamilton*, SK Martin, J Ekins, MC Rodriguez than 28 d after the cessation of treatment to return to preinfestation levels, possibly due to continued antigenic stimulation from mites or Eastern Virginia Medical School, SoBran Bioscience, Inc, Norfolk, VA dead mite debris. The results show that D. musculi is an important pathogen to exclude in allergy or atopy studies where IgE levels are Newer technologies such as infrared (IR) thermometers (IT) and IR important. imaging cameras (IC) offer an alternative to invasive techniques such as intrarectal thermometers and intracorporal implants to measure body temperature (BT) in mice. Knowing that IR technologies measure heat dissipation from skin, we set to measure if the adipose layer of obese mice serves as insulation, affecting the temperature

680 Abstracts of scientific papers 2019 AALAS National Meeting

P278 Mite Burden and Immunologic Response to Demodex early loss of RPE, thinning or dysplastic change in choroid, and musculi in Swiss Webster, BALB/cAnNCrl, and C57BL/6NCrl Mice progressive degeneration of the retinal outer nuclear layer (ONL). The ganglion cell and nerve fiber layers were better preserved and M Gugel*1, RJ Ricart Arbona2,1, K Daniels3, R Gardner3, W Boteler5, sclera remained unaffected. Immunohistochemistry confirmed initial CL Perkins4, KS Henderson4, N Lipman2,1 presence of some RPE followed by rapid loss in FCRA lesions. FCRA lesions in Albino rats show progressive degeneration of retinal layers 1Tri-Institutional Training Program in Laboratory Animal Medicine within the affected region with concurrent thinning of the underlying and Science, Memorial Sloan-Ketting Cancer Center, Weill Cornell choroid or, rarely, dysplastic thickening of the choroid. Minimal to no Medicine, The Rockefeller University, New York, NY; 2CCMP, lateral expansion of the lesions was detected throughout the 6-mo Memorial Sloan Kettering Cancer Center, Weill Cornell Medicine, observation period. An understanding of the natural progression of New York, NY; 3FCCF, Flow Cytometry Core Facility, New York, FCRA lesions is important to identify the possibility of progression NY; 4Charles River Laboratories, Wilmington, MA; 5Immuno Probe, associated with administration of pharmaceutical agents during Frederick, MD routine toxicology safety testing in affected rat strains.

Historically, detection methods for Demodex musculi were unreliable P280 Evaluation of High Doses of Buprenorphine for Greater and testing was rarely performed as its presence in laboratory mice Effectiveness and Duration of Analgesia in Mice went under recognized. While infestations are clinically inapparent in most mouse strains, D. musculi burdens are increased and clinical MC Terzi*, T Coksaygan, J Duggan, R Sanchez, A Panda, I Tatarov, LJ signs have been reported in select immunodeficient mouse strains. DeTolla However, the parasite’s impact on the immune system is unknown. Current testing methods include fur pluck, deep skin scrape, biopsy, Pathology, University of Maryland School of Medicine, Baltimore, and PCR assay. We characterized and correlated mite burden with MD PCR copy number and immunologic changes in naïve NOD.Cg- PrkdcscidIl2rgtm1Wjl/SzJ (NSG; mite burden only), Crl:CFW (Swiss Buprenorphine is a partial opioid agonist commonly found in rodent Webster), C57BL/6NCrl, and BALB/cAnNCrl mice following surgery protocols as a postsurgical analgesic. Traditionally, small exposure to Demodex-infested NSG mice. Age-matched mice of each doses (0.05–3.0 mg/kg SQ) have been used which must be strain (n=5) were euthanized 14, 28, 56, and 112 d postexposure to a administered every 6 to 12 h. A more convenient dosing schedule Demodex-positive or Demodex-negative NSG mouse. Mite burden may increase compliance, thereby increasing the welfare of the was determined by PCR and skin histopathology; B cell, CD4, and rodents after surgical manipulation. Simbadol is a buprenorphine CD8 cell counts were evaluated using flow cytometry; complete injection for cats, which, unlike other sustained-release blood counts were performed, and serum IgE levels were measured buprenorphine pharmaceuticals, contains no slow-release element by ELISA. Mite burdens in NSG mice continued to increase while and simply increases the concentration of buprenorphine at the time they plateaued at a considerably lower (100-fold) level in of administration for a 24-hanalgesic duration. This study attempts to immunocompetent strains. Most infested animals developed diffuse determine if higher doses of buprenorphine can extend the duration alopecia by 112 d postexposure. There was no correlation between of analgesia in mice. C57BL/6J and CD1 mice were evaluated for PCR copy numbers in immunecompetent strains, likely due to baseline nociceptive response by exposing them to a hot plate inherently low mite burdens. However, in NSG mice there was a stabilized at 50° C and measuring the time between exposure and significant correlation (R2=0.75). No clinically significant hematologic response (a vocalization, a jump, or a hind paw lick). The following abnormalities or alterations in immunophenotype were evident in day, the same mice were treated with a single dose of 1 of 9 doses of any of the strains throughout the experimental timepoints. BALB/c buprenorphine (0.05, 0.1, 0.5, 1, 2, 3, 5, 7, 10 mg/kg), then challenged and C57BL/6 mice had significant increases in IgE levels starting 28 on the hot plate at 30 min, and at 4, 8, 24, 48, and 120 h (n=10 for each ds postexposure. These findings align with the inherent group; 5 males, 5 females). The results of our study suggest that for immunophenotypes of each strain and demonstrate a need acute, A-δ fiber nociception, doses of 2 mg/kg or higher may be for Demodex musculi surveillance in immunocompetent mouse necessary for appropriate analgesia; at doses of 5 mg/kg, C57BL/6J colonies as the immune perturbations that result from infestation mice may experience prolonged analgesic duration; and, at doses may impact the use of infested mice in select studies. between 5 and 10 mg/kg, the mice trend toward hyperalgesic behavior at time points as far as 120 h postinjection. The main side P279 A Prospective Longitudinal Optical Coherence Tomography effect observed was hypoactivity in doses above 5 mg/kg. To treat Imaging Study Demonstrates Focal Chorioretinal Atrophy Is a Type A-δ fiber-associated pain, we recommend using a dose of 1–3 Progressive Disease in Sprague-Dawley Rats mg/kg in CD1 mice for appropriate analgesia; 2–3 mg/kg for appropriate analgesia in C57BL/6J mice; and 5 mg/kg in C57BL/6J MT Leahy*, J Bartoe, R Boyd, K Nelson, T Vihtelic mice to extend the duration of analgesia for up to 48 h. Further studies are necessary to determine whether these doses are Surgery and Efficacy, Ophthalmology, Charles River Laboratories, appropriate to treat type C fiber-associated pain, as well as to Kalamazoo, MI determine the significance of potential side effects. We feel the risk of very limited side effects are outweighed by the benefits of Focal chorioretinal atrophy (FCRA) affects albino rat strains with enhancement and duration of analgesia at the higher doses of 1–5 incidence of 0.5-2%, yet sparse data are available defining if the mg/kg. lesions progress and at what rate. As FCRA is an important background observation in routine preclinical toxicology studies, a P281 Arterial Access for Serial Coronary Intervention: Where to Go prospective longitudinal study was designed to evaluate progression. and When Our study aimed to see if FCRA does progress with age in Spragye- Dawley Rats. Six male and six female Sprague-Dawley rats were A Corn, ME Burke*, D Ordanes, G Yi selected after FCRA was diagnosed via indirect ophthalmoscopy during screening of populations purchased for unrelated toxicology Veterinary Staff, Cardiovascular Research Foundation, Orangeburg, studies. Under sedation, animals were imaged via optical coherence NY tomography (OCT) monthly until termination. Two males and 2 females were euthanized at time points 2, 4, and 6 mo. Eyes were Serial arterial vascular access poses a clinical challenge in coronary enucleated and prepared for pathologist evaluation of hematoxylin- interventional preclinical studies. A coronary stenting project eosin staining and RPE65 immunolabelling. In FCRA lesions, required weekly arterial vascular access for 5 wk followed by progressive thinning of outer retinal layers was detected via OCT monthly access for a maximum of 10 procedures via the carotid or imaging throughout duration of the study. Histopathology showed femoral arteries. For safety reasons, each vessel was cannulated no

681 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 more than 3 times. It was hypothesized that rotation between access immune function exist between species, which makes translating sites would be more time efficient with less postoperative results difficult. To address this issue, the humanized mouse model complications than reaccessing the same vessel consecutively. Twelve was developed to more closely mimic the response of a human Yucatan swine were split into 2 groups: A and B. In group A, the immune system in mice. The humanized mouse is created via a accesses rotated among the carotid and femoral arteries to maximize multistep process, which begins with irradiation of an time between cannulating the same vessel, which would potentially immunodeficient mouse to deplete any native immune cells. The promote healing and revascularization of the vessel. Alternatively, in mice are then injected with human CD34+ hematopoietic cells, which group B, each access site was used consecutively to minimize time develop into the various cellular components of the human immune between reaccesses, which could potentially allow the surgeon to system. Currently, 5 mice are irradiated simultaneously in a shoebox maximize the use of the vessel before formation of significant scar pen (10” x 6” x 4.5”) and can roam freely. As mice are communal tissue or thrombus in the artery. Group B had slightly shorter average animals, this mobile freedom frequently leads to huddling behavior, surgical times (35.5 min compared with 38 minutes), but the animals which potentially impacts the uniformity of the radiation dose given experienced more postoperative complications with the ventral neck to each mouse. We hypothesized that mice irradiated in individual incision. The animals in group A had fewer postoperative partitions would prevent them from roaming freely, which would complications due to allowance for incision site healing. Carotid improve the uniformity of radiation doses and therefore engraftment access via cutdown was both time efficient and associated with fewer percentages. We tested this by creating a 10-partition barrier inside intraoperative complications when compared to percutaneous the pen. Naïve 4-wk-old NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV- femoral access. However, subsequent carotid accesses became IL3,CSF2,KITLG)1Eav/MloySzJ (SGM3) mice were irradiated at 100 increasingly difficult due to repeated ligations. Benefits from the cGy and divided into 2 experimental groups based on irradiation shorter average surgery time in group B did not outweigh the time method: roam free (RF, n=15) and individual partitions (IP, n=10). spent by increased postoperative care of incisional complications nor These groups were repeated in 2 CD34+ cell donors (donor 5714 and the potential discomfort the animals experienced. Overall, the pattern donor 5722). At 4 h postirradiation, CD34+ cells were thawed, used by group A, vascular access rotation, was the superior suspended in PBS and injected intravenously into the tail vein. At 10 permutation for serial arterial accesses. wk postengraftment, blood was collected from the retroorbital sinus and analyzed by flow cytometry for the following markers: hCD45, P282 Characterization of a Diet-induced Nonalcoholic hCD19, hCD3, hCD33, and mCD45. For both CD34+ donors, there Steatohepatitis Mouse Model were no significant differences in engraftment levels in hCD45, hCD19, hCD3, or mCD45 (P > 0.05). However, for donor 5722, there MM MacBride*, M Gupta, J Richardson, MM Hanson was a significant difference in engraftment in hCD33 (RF=19.4%, IP=15.5%, P = 0.020), though hCD33 was not significantly different in Taconic Biosciences, Hudson, NY donor 5714. In conclusion, the use of individual partitions when irradiating mice does not affect human engraftment levels, though Nonalcoholic fatty liver disease (NAFLD), along with its more severe there is some donor-specific evidence to suggest that myeloid manifestation, nonalcoholic steatohepatitis (NASH), is a growing engraftment is impacted. public health threat. As there are currently no approved therapeutics to treat NASH, the need for preclinical animal models for drug P284 Large Animal Models for Auditory Research discovery is great. A commonly used diet-induced NASH model consists of C57BL/6 mice fed a high-fat, high-fructose, high- MJ Williams-Fritze*1, C Sage2 cholesterol diet with a large proportion of a trans fat. With the FDA ban on trans fats, this diet is no longer available. We thus sought to 1Veterinary Services, CBSET Inc, Lexington, MA; 2CILcare, Inc., validate whether a replacement diet using palm oil in place would Lexington, MA induce NAFLD/NASH in C57BL/6NTac mice. Pilot production was aimed at identifying the best parameters for production of a Auditory studies, including those evaluating ototoxicity and consistent model. C57BL/6NTac male mice were started on the diet pharmacokinetics (PK), are typically performed using rodent models. at 5 or 6 wk of age. Evaluation of weights just prior to diet Some studies, however, may require a larger species with middle and administration compared to weights 17 wk later identified a inner ear anatomy more similar to humans. Currently, there is a minimum weight threshold (15 g) required for best response to diet paucity of literature describing surgical, procedural auditory in terms of weight gain (n=93). Compared to C57BL/6J mice started techniques and proper and sensitive readout in large animal species. on the trans fat diet at 12 wk of age (n=6 per time point), Our goal was to determine if large animal species, such as swine and C57BL/6NTac mice on the palm oil replacement diet (n=8 per time sheep, could be reliably used for auditory studies. Here, we report point) gained weight more quickly and had similar liver weights. successful procedures such as transtympanic injection (TT), a surgical Both groups had similar blood glucose and non-esterified free fatty approach to reach the middle ear space and recording of acoustic acid levels, but the C57BL/6NTac had reduced variance compared to auditory brainstem response (aABR) in large animal species. the C57BL/6J group. Additionally, the C57BL/6NTac mice had Evaluation of the external auditory canal (EAC) anatomy to the higher transaminases, LDL, and triglycerides after 16 wk on diet. At tympanic membrane (TM) was performed in 8 sheep (3 live animals 16 wk on diet, C57BL/6NTac mice (n=10) were obese, with enlarged, and 5 cadaveric heads) using endoscope. Using endoscopic fatty livers. No evidence of liver tumors was seen by gross examination we demonstrate that sheep are better suited for visualization. At 38 wk on diet, C57BL/6NTac mice (n=8) displayed transtympanic injection with an EAC/TM closer to human much higher body weight, liver weight, cholesterol, LDL, and architecture compared to swine (EAC and TM at 90 angle). Although transaminases compared to control mice on chow diet (n=4). The both swine and sheep have anatomical features of the middle/inner data support use of the palm oil diet as an appropriate diet for ear similar to humans, sheep can be used for chronic PK studies, as induction of NAFLD/NASH in C57BL/6NTac mice. the TM is easily accessible. For swine, the surgical approach through the middle ear to reach the TM is complex, and thus, swine are not P283 Irradiating SGM3 Mice in Individual Partitions Does Not recommended for survival studies. For medical devices, such as Impact Engraftment Success of Human CD34+ Hematopoietic Cells cochlear implants, we determined that both species could be used using a similar surgical approach (~6-8 cm incision anterioventral to M Palomares*, AS Yokoyama, P Sproul, M Hente, P Kaur the base of the ear with blunt dissection and drilling into the middle ear space). In addition, we successfully calculated the aABR In Vivo Services, The Jackson Laboratory, Sacramento, CA threshold from both swine (n=4) and sheep (n=3) under different anesthesia protocols (isofluorane by inhalation and/or propofol CRI) The mouse is commonly used as an in vivo model for studying using a homemade multichannel ABR system with or without human immunology. However, fundamental differences in cellular electrode brain implantation (i.e. SQ sensor placement). For both

682 Abstracts of scientific papers 2019 AALAS National Meeting

species the aABR threshold is not affected by anesthesia protocol for treated with 10% chlorhexidine solution [spray] had final scores of 0 all frequencies tested and our system is sensitive enough to be used and 1, sodium hypochlorite (bleach) solution (0.005%) [spray] had with subcutaneous electrode alone. In summary, our studies show final scores of 2 and 1, standard triple antibiotic ointment had final that large animal species can be successfully used in standard scores of 2 and 3, and ophthalmic triple antibiotic ointment had final auditory testing procedures. scores of 3 and 3. Preliminarily, this pilot study data suggests that dermatitis can be treated most efficiently by 10% chlorhexidine P285 Comparison of Immunogenicity and Safety Outcomes of 2 solution [spray]. Malaria Vaccines Tested in Rhesus Macaques (Macaca Mulatta) of Indian And Chinese Origin P287 Intestinal Colonization of Genotoxic Escherichia coli Strains Encoding Colibactin and Cytotoxic Necrotizing Factor in Small ML Martin*1, A Bitzer2, AJ Schrader1, KH Soto2, FA Khan2, MD Mammals Langowski2, Z Beck3,4, GR Matyas3, AH Batchelor2, S Dutta2 N Fabian*, A Mannion, Y Feng, CM Madden, JG Fox 1Veterinary Service Program, Walter Reed Army Institute Research, Silver Spring, MD; 2Laboratory of Structural Vaccinology, Walter Division of Comparative Medicine, Massachusetts Institute of Reed Army Institute Research, Silver Spring, MD; 3Military HIV Technology (MIT), Cambridge, MA Research Program, Walter Reed Army Institute of Research, Silver Spring, MD; 4Henry M Jackson Foundation, Rockville, MD Escherichia coli encoding colibactin (pks), cytolethal distending toxin (cdt), and hemolysin-associated cytotoxic necrotizing factor (cnf) are Indian-origin (IO) Macaca mulatta are the current model of choice for associated with various intestinal and extraintestinal diseases in malaria vaccines studies. However, since 1978, there has been a ban humans and animals. Small mammal pets, many of which are on importing IO rhesus macaques making research with these popular laboratory animal species, are seldom evaluated for animals difficult due to the increase in demand. Chinese-origin (CO) genotoxin-encoding E. coli, and thus the prevalence of such strains is rhesus macaques are easily obtainable and are morphologically unknown. We hypothesized that small mammal species are colonized similar to IO. Currently, the most promising malaria vaccine target with genotoxic E. coli strains. The objective of this study was to the Circumsporozoite Protein (CSP). We developed 2 vaccines based isolate and characterize genotoxin-encoding E. coli from healthy and on CSP. A recombinant soluble nearly full-length CSP vaccine and an ill small mammals examined at a veterinary clinic, 2 animal adoption epitope-based vaccine which uses the tobacco mosaic virus (TMV), centers, and an academic research institution. E. coli isolates were both vaccines are combined with the Army liposomal formulation cultured from fecal samples for molecular and biochemical (ALFQ). We hypothesized that CO and IO rhesus can interchangeably characterization. A total of 65 pets, including mice, rats, rabbits, be used as animal models to distinguish between malaria vaccines. guinea pigs, and hedgehogs, were screened. Additionally, a 4-y-old Two groups of CO and IO monkeys (n=6) received either the TMV- female Dunkin-Hartley laboratory guinea pig was evaluated after CSP+ALFQ or the CSP+ALFQ vaccines intramuscularly at months having acute weight loss, polyuria/polydipsia, and diarrhea. 0-1-2. Blood samples were taken at days 1, 3, 7, 14, and 28. Both Twenty-seven E. coli isolates were obtained from 25 animals. Twelve models reproduced a mild skin reaction after the vaccination with no of the 27 isolates (44.4%) were PCR-positive for elevations in renal and liver function tests. Both models reproduced a the pks genes clbA and clbQ. Two isolates (7.4%) were PCR-positive transient increase in CK and WBC counts following vaccination. Both for cnf. All isolates were PCR-negative for cdt. The laboratory guinea models predicted the superiority of TMV vaccine over CSP barring pig had E. coli isolated from feces; however, the isolate was negative minor differences in their ability to detect differences in avidity and for all 3 genotoxins. All genotoxin-encoding isolates belonged to the CO rhesus induced more functional antibodies by in an Inhibition of pathogen-associated phylogenetic group B2. Representative liver stage development assay. We conclude that Chinese origin genotoxin-encoding isolates had serotypes previously associated rhesus macaques display similar immunological responses to Indian with clinical disease in humans and animals. Isolates origin rhesus macaques when vaccinated with malaria vaccines and encoding pks or cnf induced megalocytosis and cytotoxicity in HeLa as such may be a valid alternate model for malarial vaccine cells in vitro. Although most isolates were obtained from healthy development. animals, 2 pet guinea pigs with diarrhea had pks-positive E. coli isolates cultured from their feces. Whole genome sequencing of 4 P286 How to Stop The Itch: Dermatitis Treatment for Mice representative isolates confirmed the presence of pks and cnf genes and identified other virulence factors associated with pathogenicity M Richburg*, M Blackshear in animals and humans. Our results indicate that small mammals are a reservoir for potentially pathogenic E. coli with zoonotic risk. Animal Research Facility, MedStar Health Research Institute, Cheverly, MD P288 Potentiation of Macrophage Response to M. tuberculosis via Vitamin A-dependent Nuclear Receptor Activation In our breeding colony consisting of various C57BL/6 derived mouse strains, we noticed that many were developing dermatitis. N Mishkin*, J Dilisio, B Podell, R Basaraba Traditionally, our facility would topically apply ophthalmic triple antibiotic ointment to the affected areas. However, even when College of Veterinary Medicine and Biomedical Sciences, Colorado treating the animal daily, signs of improvement took several weeks. State University, Fort Collins, CO We decided to test the efficacy of other treatments that other research laboratories used and others we had used previously, but change the The development of active tuberculosis, the leading cause of death duration, frequency, and application method. We randomly selected due to infectious disease, is often the result of known risk factors, 8 mice with dermatitis with varying degrees of severity. We then either intrinsic or extrinsic. Recently, vitamin A deficiency has been separated the mice into 4 groups, with each group to be treated with implicated as such, however the mechanisms underlying this risk are a different substance: 10% chlorhexidine solution [spray], sodium yet to be elucidated. The aim of this study is to better understand the hypochlorite (bleach) solution (0.005%) [spray], standard triple role of vitamin A in macrophage immune function, through antibiotic ointment, and ophthalmic triple antibiotic ointment. We investigating nuclear receptors (PPARγ, RAR, and RXR) and enzymes sprayed or topically treated the mice a total of 8 times, 4 times per (RALDH1A1 and RALDH1A3) associated with vitamin A’s metabolic week consecutively, over a 2-wk period, with the substance to which pathway. Administration of agonists of these targets should lead to each group was assigned. We used dermal scoring of the typical their activation, which we hypothesize will improve the innate erythema characteristic to quantify our findings on a scale of 0-5, response to infection and consequently antimicrobial capacity. Bone where 0 is no sign of erythema and 5 is severe erythema on >75% of marrow-derived macrophages obtained from guinea pigs were used the original treated area. After the 8 treatments, the animals being as an in vitro cell model of inflammatory macrophages preferentially

683 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 infected by M. tuberculosis in vivo. Through quantification of protein melanoma and MC38 colon adenocarcinoma) with 2 different expression by western blot and gene expression analysis using qRT- concentrations (1x105 and 5x105) in young (7-wk-old) and aged PCR, nuclear receptor targets and immune parameters were assessed (78-wk-old) female C57BL/6J mice, and evaluated tumor growth in context of administration of retinoic acid, the bioactive form of kinetics. Mice were randomly assigned (n=10/group) and 1x105 or vitamin A, and a known PPARγ agonist – rosiglitazone, or 5x105 cells in PBS were injected subcutaneously in their right flanks. combination of both in uninfected macrophages and those infected Mice were monitored daily post cell injection and trilogies (clinical with Mtb. Bacterial growth in the infected cells of these same observations, bodyweights, and tumor measurements) were treatment groups was measured through quantification of colony conducted 3 times per week once the tumor volume reached around forming units (CFUs), and was compared to an MTT viability dye to 50mm3. In both syngeneic models, tumor growth happened at a more assess impact on antimicrobial activity. Initial results show reduced rapid pace in the young mice than in the aged mice for those given expression of proinflammatory cytokines and improved cellular 1x105 cells and more young mice had to be euthanized before study viability in all treatment groups, suggesting increased macrophage terminus due to tumor volumes reaching endpoint (2000mm3). immune response through vitamin-A dependent pathways. Tumor growth in the groups given 5x105 cells was relatively the same across young and aged mice, suggesting rapid tumor growth P289 Urinary Protein Analysis in Mice Lacking the Major Urinary hindered the difference between young and aged mice. Young mice Proteins gained body weight over the course of time, while aged mice lost body weight, suggesting cancer cachexia happens in aged mice. O Suzuki*, M Koura, K Uchio-Yamada, M Sasaki, Y Doi Across all studies, tumors in the aged mice grew less rapidly than they did in their younger counterparts, only in those inoculated with Laboratory of Animal Models for Human Diseases, National lower (1x105) tumor cells. This data verifies the need for aged mice to Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, , be included in cancer research because the tumor kinetics are Japan different in young and aged mice. The observed differences could potentially bridge the gap between successful preclinical cancer In mice, major urinary proteins (Mups) are synthesized in the liver, studies and successful clinical trials. and a considerable amount of the proteins are exreted into the urine. Therefore, even healthy mice exhibit proteinuria, unlike healthy P291 Evaluation of a Diagnostic A1C Report for Adult Rhesus humans, which often makes it challenging to use mice as models for Macaques (Macaca mulatta) research in human urological diseases. To circumvent this problem, we produced Mup knockout (Mup-KO) mice by gene editing with a PO Mills*1, K Nestor2, J Condrey1, RA Howard1, B Herrod1, E similar strategy reported previously and examined the composition Schlosser2, G Ray3 of their urinary proteins. We removed a ~2.2-megabase-long cluster of Mup genes on chromosome 4 in mice (C57BL/6N) by TAKE 1National Center for Emerging and Zoonotic Infectious Disease, method with Cas9 proteins and 2 guide RNAs (gRNAs) targeting Centers for Disease Control and Prevention, Atlanta, GA; 2Charles both ends of the cluster. We selected genetically modified mice by River Laboratory, Atlanta, GA; 3Baycom Diagnostics, Talahassee, FL PCR detecting removal of the gene cluster. We measured urinary protein concentrations in Mup-KO and wild-type mice by a The natural development of Type 2 diabetes mellitus in the adult fluorescence-based protein quantitation kit and urine test strips. We rhesus macaque is a common clinical occurrence. Traditional also examined urinary protein compositions with SDS-PAGE and methods to assess the glycemic state over a prolonged interval (>2 2D-PAGE. We obtained pronuclear embryos (C57BL/6N) by in vitro wk) in adult rhesus macaque require either stressful (physical or fertilization and introduced Cas9 protein/gRNA complexes into 92 chemical) restraint, or large amounts of blood collection per sample. pronuclear embryos. After overnight in vitro culture of the embryos, The A1Care report is more specific in detection and screening of Type we transferred 87 two-cell embryos into uterine foster mothers (ICR). 2 diabetic mellitus adult nonhuman primates than the commercial We finally obtained 17 weaned pups. The Mup-KO mice established laboratory A1c test. Indoor-housed adult male and female Rhesus from them by reproduction and selection with genotyping PCR were macaques (Macaca mulatta; n= 8: age 8-10-y-old) were randomly healthy and fertile. The urinary protein concentration in Mup-KO selected for clinical screening of Type 2 diabetes mellitus. Fasting mice (17.9 ± 1.8 mg/dL, mean ± SD, n=3) was significantly lower (P < blood glucose was concurrently collected from each animal, 0.001) than that of wild-type mice (73.7 ± 8.2 mg/dL, n=3). Protein including 0.2mL of whole blood for diagnostic A1C (using cation measurement with urine test strips also showed the same tendency. exchange high-performance liquid chromatography) and 3mL of Protein analysis confirmed that Mup-KO mice had no Mup proteins EDTA whole blood for the commercial laboratory A1C test (using in their urines, in contrast, that a large amount of Mup proteins, latex immunoagglutination inhibition). One animal tested positive approximately 20 kDa, appeared in urines of wild-type mice. Loss of for Type 2 diabetes with the A1Care test, whereas commercially urinary Mup’s in MupKO mice was also confirmed. These results available fructosamine and commercially available A1C tests showed indicate that Mup-KO mice produce urines more similar to human a false negative. The positive results were confirmed via fasting urines than wild-type mice do, and therefore, Mup-KO mice would blood glucose and intravenous glucose tolerance tests. Results be useful as a model for human urinalysis. We are planning to indicate the A1care report for nondiabetic adult rhesus macaques distribute the mice through our laboratory animal resource bank. ranges from 4% to 5.7% binding of A1C to hemoglobin. This study supports the usage of A1Care for the detection of diabetes mellitus in P290 Comparing Tumor Growth Rate in Young Compared with adult rhesus macaques. The A1care report allows for refinements to Aged C57BL/6J Syngeneic Model these aforementioned procedures so that the test can be conducted cage-side without the added stress of sedation and requires only a P Collier*, J Yang, M Cheng 200uL blood samples.

The Jackson Laboratory, Sacramento, CA P292 Consequences of Chronic Exposure of Rainbow Trout (Onchorhynchus mykiss) to Subinhibitory Levels of the Antibiotic Aging has been directly correlated to the likelihood of developing Sulfamethoxazole cancer. Numerous preclinical cancer studies fail once they reach clinical trials and one contributing factor could be that most tumor RN Labitt*1,2, H Marquis2 research is done in young, healthy mice while the actual cancer population tends to be older. The purpose of this study is to begin 1Center for Animal Resources and Education, Cornell University, establishing tumor models in aged mice, which would better Ithaca, NY; 2Microbiology and Immunology, Cornell University, represent cancer patients. In order to do this, we established mouse Ithaca, NY syngeneic tumor models using two different cell lines (B16-F10

684 Abstracts of scientific papers 2019 AALAS National Meeting

It is well established that natural waters are contaminated with Parvovirus (MKPV), is a single stranded, non-enveloped DNA virus antibiotics, in the United States and internationally. These categorized as a member of a currently unclassified Parvovirus contaminants are in large part due to the excretion of unmetabolized subfamily. MuCPV is genetically and antigenically distinct antibiotics in urine and feces, improper disposal of unused from Protoparvoviruses (e.g. MVM, MPV). MuCPV has recently been antibiotics, and the variable effectiveness of waste water treatment linked with the development of kidney disease in laboratory mice, plants in eliminating antibiotics. We have a negligible knowledge of specifically infection in immunodeficient animals leading to severe the consequences of chronic exposure to subinhibitory levels of kidney disease and renal failure. Immune competent strains infected antibiotics on aquatic life. However, in vitro experiments have shown with MuCPV are less affected by the virus. A recombinant MuCPV- that, at sub-inhibitory levels, antibiotics are very potent signaling VP1 protein expressed in baculovirus/insect cells was purified and molecules influencing bacterial gene expression, facilitating used to develop a serologic assay for detection of antibodies in mutagenesis and horizontal gene transfer, and modulating bacterial mouse serum. Infected SF+ insect cells supernatant was metabolism and virulence. We hypothesized that chronic exposure to ultracentrifuged to pellet protein aggregates which were then further subinhibitory levels of antibiotics would lead to the emergence of purified by OptiprepTM gradient. Potency of the purified protein was antimicrobial resistance within the fish intestinal microbiota. To test determined by enzyme linked immunosorbent assay (ELISA) and this hypothesis, we elected to use sulfamethoxazole (SMX), the most multiplexed fluorometric immunoassay (MFIA®). Purity of the common antibiotic detected in natural waters. Initially, SMX purified 57kD VP1-protein was assessed by staining of SDS-PAGE minimum inhibitory concentration (MIC) and no observed effect gels and western blot (WIB) analysis.Screening of sera from pet shop concentration (NOEC) were determined for the intestinal microbiota and vendor mice by PCR and serology showed a prevalence of >15% of juvenile rainbow trout. Fish were then exposed for 6 mo to in the population. Virus detection in different tissues including subinhibitory concentrations of SMX. Acquisition of antimicrobial lymph nodes, kidney and feces varied based on the age of the animal. resistance was assessed phenotypically by quantifying the ratio of None of the 300 tested sera showed positive results thus resistant to total colony forming units from the intestinal microbiota. demonstrating a 100% specificity of MuCPV-VP1 MFIA assay. Purity Exposure of fish to the NOEC, a dose approximately 80-fold lower of the MuCPV antigen was further tested by screening heterologous than the MIC, resulted in a 10,000 fold increase in resistant colonies. sera positive for other infectious agents (i.e. LCMV, MPV, MTLV, etc.). Susceptible and resistant colonies were archived and will be assessed Results showed no cross-reactivity with non-MuCPV antibodies. In genotypically for the presence of known SMX resistance genes, sul1- summary, MFIA® assays developed using MuCPV-VP1 antigen 3, and acquisition of single site mutations in the folp gene, whose demonstrated good sensitivity and specificity for detection of product is the target of SMX. Results from this study indicate that MuCPV sera antibodies and can be used for routine screening of chronic exposure to very low levels of antibiotics contribute to the mice colonies. development of antimicrobial resistance in aquatic animals and potentially their environment. P295 Comparing Subcutaneous with Mammary Fat Pad Implantation on Growth Kinetics and Histology of PDX Breast P293 We Are What We Eat: The Impact of Fasting and Bedding on Cancer Models Blood Glucose and Regional Brain Uptake of 18F-FDG R Banzon*, M Tewodros, D VanBuskirk, K Draheim, M Cheng R Speedy*, KM Eldridge, AA Bedwell, P Territo Innovative Product Development, The Jackson Laboratory, Imaging and Radiology, Indian University, Purdue University Sacramento, CA Indianapolis, Indianapolis, IN Preclinical oncology studies frequently use patient-derived Imaging of glucose metabolism in rodent models has been used for xenograph (PDX) tumor models and therapy efficacy is evaluated by nearly 30 y for cardiovascular, cancer, and neurological disorders. To measuring changes in tumor volume. Subcutaneous (SQ) ensure that glucose is transported into brain tissue, animals are implantations of tumors are most widely used in oncology studies fasted for 12-18 h prior to reduce the direct completion of due to ease of technique and tumor accessibility for caliper 18F-fluorodeoxyglucose (18F-FDG) with its native substrate across measurements. However, SQ tumors are then growing in a different GLUT1 transporters. Recent observations in our laboratory indicate environment compared to the tumor model. Conversely, orthotopic that mice resort to foraging for residual food, eating their bedding, or implantations provide tumors with a similar microenvironment for enrichment left in cage during fasting periods. Therefore, we growth, but require surgical techniques which increase the amount of hypothesize that animal bedding would alter blood glucose levels resources, time, and skill needed to perform these studies. Currently, and thus impact 18F-FDG uptake. To test this, 16 male and 16 female the standard method for an orthotopic breast cancer involves C57BL6/J mice were divided into 4 animals/group/sex then placed surgically clearing the mammary fat pad (MFP) prior to engrafting into clean cages containing an aspen wood bedding product, the cells or tissue. In this study, we developed a nonsurgical corncob, or paper pellet bedding the day before PET scanning and approach where the PDX tissue is engrafted directly into a were fasted for 18 h. On scan day, blood glucose values were taken noncleared MFP using a method similar to SQ engraftment. This and then animals were injected with 18F-FDG IP. After a 45-min study compares the effect of SQ and MFP implantations on growth uptake period, mice were anesthetized with isoflurane and placed on kinetics and tumor histology using several different preestablished the scanner, imaged for 15 min, and then euthanized for postmortem breast cancer PDX models with doubling times ranging from 7-10 d autoradiography. Results showed that there is a significant sexual to 40-55 d. For each model, NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) dimorphism in blood glucose levels and regional brain PET uptake, mice were put under anesthesia and we implanted 40ul of minced and a bedding dependent uptake of brain 18F-FDG in these mice. tumor tissue SQ in 5 mice using a 14g needle, 10 mice in the MFP These suggest these factors should be not only reported but with 18g needle, and 10 mice in the MFP with 14g needle for each controlled for all imaging studies where glucose is being monitored. tumor model. Clinical observations and tumor volumes were recorded 2 to 3 times a week until mice met endpoint criteria. There P294 Development of a Recombinant Antigen for Serological was no statistical difference in latency, doubling time or engraftment Detection of Antibodies Against Newly Discovered Murine rates between implantation methods in each of the tumor models Chapparvovirus tested. Additionally, there were no major histological differences seen with H&E staining. SQ implantation and both noncleared MFP RK Dhawan*, ML Wunderlich implantation approaches required similar resources, time, and skill level. In the models tested, all implantation methods demonstrated BioAssay Services, Charles River Laboratories, Wilmington, MA similar engraftment rates and growth kinetics. Therefore, the noncleared MFP technique uses few resources, provides a short Murine Chapparvovirus (MuCPV), also referred to as Mouse Kidney recovery time, and is a minimally invasive procedure that mimics the

685 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 natural environment. Future preclinical oncology studies using breast lengths ≥ 1 inch for cervical IM injections in Yucatans and ≤ 1 inch for cancer PDX models should consider using the non-cleared MFP IM hindlimb injections in Göttingens and Yorkshires provided engraftment technique. accurate delivery into targeted muscles. IM injection site dimensions were more variable than those of SC sites. This may be due to IM P296 Daily Ketamine Administration Does Not Alter the Gut injections occasionally being deposited within intramuscular fascial Microbiome of CD-1 Mice planes, allowing the injecta to spread extensively between muscle groups. R Dashek*, S Gerb, A Ericsson, CL Franklin P298 A Novel Method for Isolating Exosomal miRNA from Mouse University of Missouri, Columbia, MO (Mus musculus) Feces

The gut microbiota (GM) is a critical player in the physiology of S Manning*1, Y Imamura Kawasawa2 animals in both health and disease. Many factors have been shown to play a role in modulating the composition of the GM of animals used 1Comparative Medicine, Penn State Hershey Medical Center, in research including, diet, husbandry practices, and certain Hershey, PA; 2Departments of Pharmacology and Biochemistry and experimental treatments. In turn, the GM has been shown to Molecular Biology, Institute for Personalized Medicine, Penn State influence the outcome of various experimental measurements. Hershey Medical Center, Hershey, PA Currently there is a lack of research examining the effects of various anesthetic agents on the GM of mice. The hypothesis of this study is Exosomes, a subtype of extracellular nano-sized vesicle, have been that daily ketamine administration significantly alters the GM shown to mediate extracellular communication via transport of composition of CD-1 mice. Twenty adult female CD-1 mice were split various biomolecules including coding and noncoding RNAs, such into 2 groups: 1 group receiving daily intraperitoneal injections of 100 as microRNA (miRNA). This process impacts several physiological mg/kg ketamine daily, and 1 group receiving daily injections of an and pathological processes. In response to this, investigation of the equivalent volume of saline, both for 10 d in order to model an use of exosomal miRNA as biomarkers has grown significantly. experimental protocol involving chronic ketamine exposure. Feces While multiple methods for isolation of exosomes from samples were collected immediately before the initiation of experimental such as blood, cells, and tissues are available, little is published on protocol, and then again following the completion of the 10 days. effective methods for isolation from feces. Other published protocols Animals were weighed every other day throughout the experiment require specialized equipment such as ultracentrifuge or technically to ensure that they did not lose a significant (>10% total body weight) difficult procedures such as density gradient separation. Here we amount of body mass and to adjust the ketamine dosage as necessary. present a novel isolation method of exosomal miRNA from mouse DNA was extracted from fecal samples and amplified by PCR using (Mus musculus) feces. We propose this method to be a robust and bacterial 16S rRNA primers, and then processed for next generation standardized practice that will have an impact on this growing field. sequencing. Sequence data were analyzed using PAST 3.25 software Additionally, analysis of exosomal miRNA assists in monitoring and groups were compared using permutational multivariate mouse models of IBD in applications where relying solely on clinical analysis of variance. Analysis revealed no significant effects on the scores falls short. Briefly, fresh fecal pellets were collected from GM by daily administration of 100 mg/kg ketamine for 10 d. These C57BL/6J mice and stored in -80°C until processing. Two fecal pellets results indicate that daily ketamine administration does not (approximately 60mg) were first vortexed with steel beads with the significantly alter the GM of mice used in research. same mass of isotonic buffer, followed by centrifugation to pellet cellular contents and other debris. The supernatant was then passed P297 Intramuscular and Subcutaneous Injections in Research through a 0.2µm filter twice. This crude exosome preparation was Swine: A Comparison of Injection Site Characteristics Based on then further purified via use of polymer-based precipitation. After Needle Length and Anatomic Location confirmation of exosomes in the sample via electron microscopy, the sample was then quantified with nanoparticle tracking analysis. It S Hall*1, M Magagna2,4, J Veenstra3, K Nelson2 was found that the sample contained 8.891010 +/- 1.79109particles/ ml of exosomes, ranging from 75–150 nm in size. Purified exosomes 1Pathology Services, Charles River Laboratories, Mattawan, were then lysed and then the miRNA was purified via acid phenol- MI; 2Pathology, Charles River Laboratories, Mattawan, MI; 3Animal chloroform method. The resulting miRNA was quantified using Care, Charles River Laboratories, Mattawan, MI; 4Veterinary microchip electrophoresis. In summary, this procedure successfully Diagnostic Laboratory, Michigan State University , Lansing, MI created high-yield exosomal miRNA that can be applied to most species. Swine are an increasingly used model in toxicologic research, particularly for injection studies. Given the inherent variability in P299 Change in Diet Reverses Nonalcoholic Steatohepatitis in size between breeds, individual animal size should be considered in Choline Deficient Defined Amino Acid Diet-induced Mouse Model relation to needle length to ensure accurate delivery of injecta to the along with Improvement of Overall Liver Function intended anatomic site. Three Yucatan minipigs, two Göttingen minipigs, and two Yorkshire cross domestic pigs (7 total animals) S Perrotta*, A Ambade, J Al-Beik, K Bayer, M Kirkland, N Jones, J were euthanized and injected intramuscularly (IM) and Lyons, J Morrison, J Cornicelli subcutaneously (SC) with 2 mL of dye solution. IM injections were performed in the cervical and hind limb musculature using 3 needle Discovery Safety Assessment, Charles River Laboratories, lengths (0.75, 1, and 1.5 inch). SC injections were performed in the Shrewsbury, MA axillary and inguinal regions using 2 needle lengths (0.75 and 1 inch). Injection site dimensions were measured following formalin fixation Nonalcoholic steatohepatitis (NASH) is associated with excess and serial sectioning of the sites. In all breeds, SC injection site caloric intake and metabolic syndrome. Diet modification is characteristics did not vary significantly between anatomic locations front line treatment in the clinic, yet this has not been validated or needle lengths. In Göttingens and Yorkshires, hindlimb IM in animal models. In this work, we used the choline deficient injections using 1.5-inch needles were consistently deposited around defined amino acid diet (CDAA) to induce NASH to test whether the femur, deep to the targeted muscles. In Yucatans, cervical IM a change to regular diet can reverse the NASH phenotype. Fifty- injections using 0.75-inch needles were consistently deposited in the five, 8-wk-old male C57BL/6J mice were used on study. Forty- subcutis. IM injections were generally located directly under the seven were fed CDAA diet for 26 wk. At 15 wk, liver biopsies were point of needle entry while SQ injections were offset. Yorkshires had obtained by laparotomy for NASH activity score (NAS) and blood the largest ranges in depth and width for IM injections. Göttingens samples collected for ALT and Procollagen Type III N-Terminal had the smallest ranges in all dimensions for SC injections. Needle Propeptide (PIIINP) analysis. At 18 wk, animals were distributed

686 Abstracts of scientific papers 2019 AALAS National Meeting

to treatment groups based upon ALT, PIIINP, and NAS. Groups The development of the patient-derived xenograft (PDX) mouse included Group 1 (vehicle control); Group 2 (30mg/kg PO, QD for model has revolutionized cancer immunotherapy research by fast- 8 wk); Group 3 (switch from CDAA to standard chow diet for 8 tracking the start of clinical trials. PDX mice used in preclinical trials wk); and Group 4 (maintained on regular chow diet throughout the are commonly dosed intravenously (IV), or by oral gavage (PO). study). Blood was collected via submandibular vein on d 1 (prior Recently, intratumoral (IT) dosing has been introduced as a novel to treatment), d 29, and d 57 (terminal). Histopathology performed approach to delivering compounds directly inside the tumor. In this on biopsy tissue confirmed advanced steatosis and minimal fibrosis study, NOD.Cg-Prkdcscidll2rgtm1wjllSzJ (NSG) mice were implanted after 15 wk of CDAA diet feeding, overall NAS score 3.9. ALT (P < subcutaneously in the flank with PDX material. Mice were 0.0003) and PIIINP (P < 0.02) levels were significantly elevated randomized by tumor volume (~150mm3) into 5 groups. To compared to chow fed group. Diet change resulted in lowering body determine whether needle size used disrupts the integrity of the weight within first week and was accompanied by small increase tumor surface, we evaluated both 27G and 30G needles. Mice were in food intake. It also resulted in significant lowering of ALT (P < dosed with 50 µL of saline via IT dosing every other day for 10 d. 2.2E-5) and PIIINP (P < 0.0001) accompanied by lowered liver Mice were closely examined for signs of outer tumor disruptions weight, liver to body weight ratio, and liver cholesterol. The switch (erosion or ulceration). At study terminus, tumors were collected and to chow diet significantly reduced liver triglycerides (TG) which was examined for signs of internal disruption. While neither erosions nor independently confirmed by histopathology. Pioglitazone lowered ulcerations were found during the course of the treatment, it was serum TG but did not impact the liver TG. Terminal histopathology noticed that the 27G needle is more prone to causing fluid leakage NAS scores were as follows: vehicle 6.1, pioglitazone 4.5, diet when withdrawing the needle. To determine if tumor integrity is switch 2.5, and chow control 0.1. Liver biopsy lowered the number compromised, one-point injection (compound being directed to the of required animals and allowed to use each animal as its own tumor center) and fanning injection (compound being directed across control. Our data showed that within 4 wk of switching to standard several areas of the tumor) techniques were trialed. The mice chow, all liver function tests returned to normal and by 8 wk the received the same 10-d dosing regimen, with 2 groups receiving one- NASH phenotype was reversed, thus proving that diet change point injections, and 2 groups receiving fanning injections (with 27G recommended in clinics is also effective in the CDAA diet-induced or 30G needles, respectively). At study terminus, the mice were model of NASH. euthanized and tumors were collected and fixed in formalin for paraffin embedding. Due to the difficulty of moving the needle P300 Progressive Aortic Cuff Inflation as a Porcine Model for Left within the tumor during compound administration, we determined Ventricular Pressure Overload Heart Disease that the one-point technique is ideal for smaller and rounder tumors, while the fanning technique is preferred for larger, multi-focal- CD Fisher*1,2, TL Jones1,2, A Moore2, H Doviak3, M Poole1, S Barlow1,2, merged or oval-shaped tumors. In summary, repeated 27G or 30G F Spinale2 needle insertions does not appear to cause disruption at the tumor surface. And while some risk of ulceration or erosion due to the 1Department of Laboratory Animal Resources, University of South chosen compound still remains, the ideal administration technique Carolina, Columbia, SC; 2Cardiovascular Translational Research for IT dosing varies depending on the size and morphology of the Center, University of South Carolina School of Medicine, Columbia, tumors. SC; 3Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville, VA P302 Detection of Lymphocytic Choriomeningitis Virus by Exhaust Air Dust in IVC Racks Heart failure, a common global cause of death, frequently develops after a prolonged period of hypertension due to excessive load to S Durand*1, D Schaefer2, P Hardy3 the left ventricle. This form of left ventricular pressure overload (LVPO) causes a form of heart failure with unique features and is 1Veterinary and Professional Services, Charles River, L’arbresle defined as heart failure with a preserved ejection fraction (HFpEF). Cedex, France; 2LASC, University of Zurich, Zurich, However, therapies to treat HfpEF have not been forthcoming Switzerland; 3Allentown, Bussy-Saint-Georges, France due to a lack of animal models, particularly relevant large animal models, which recapitulate progressive LVPO and the HFpEF Whereas numerous prevalent infectious agents were demonstrated to presentation. Accordingly, our group has developed a porcine model be efficiently detected with exhaust air dust (EAD) samples, this was of progressive LVPO and HFpEF progression using an implantable not yet established with any zoonotic agents. It was important to hydraulic system allowing for sequential increases in LVPO. This know for safety and biosecurity reasons if this innovative requires a surgical team to perform the aortic surgery in Yorkshire environmental sampling system allowed the detection of zoonotic pigs, provide pre and post-operative management, and perform agents. Experimental infection of mice with lymphocytic serial assessment of left ventricular function using echocardiography. choriomeningitis virus (LCMV) in biosafety level (BSL) 2/3 Recently completed studies over a 5-wk period with sequential LVPO environment is frequently used as a mouse model to study acute and induction (n=15) has demonstrated a stable left ventricular ejection chronic viral infections. This made it possible to use and assess in fraction, increased left ventricular mass (hypertrophy) and functional parallel the inline EAD capture system and plenum swabbing to signs of HFpEF progression (left ventricular diastolic failure). detect the presence of LCMV in IVC racks. Two IVC racks were each Moreover, histological examination demonstrated the classical populated with 2 infected mice cages. A third IVC rack, with cages structural manifestations of HFpEF-increased extracellular matrix containing mice noninfected with LCMV, was used as negative accumulation (“fibrosis”). The specific postoperative management, control. The study mice (C57BL/6, females) were infected IV standard operating procedures, and practice standards have (2x106 pfu) with either the LCMV Armstrong strain (acute LCMV) or demonstrated this HFpEF model can be produced with few surgical with the LCMV docile Clone 13strain (chronic LCMV). The inline EAD complications and/or adverse events. Thus, this large animal model capture media were placed at the air exhaust of each rack plenum of LVPO and HFpEF can be utilized to develop new therapeutic according to manufacturer instructions. Six wk later, 3 types of strategies for this progressive and fatal disease. samples were collected: the inline EAD capture media (n=3), plenums swabs (n=3) from all racks and feces (n=6) from LCMV- P301 Validation of Different Aspects of the Intratumoral Injection infected mice cages. All samples were sent to the laboratory for real- Method time PCR testing. LCMV was detected in EAD capture media (1 positive out of 2 samples) as well as plenums (1 positive out of 2 S Yu*, M Palomares, C Simion, P Sproul, P Kaur samples) and feces (2 positives out of 4). Both LCMV infectious models were detected by EAD samples. LCMV copy number in all In Vivo, The Jackson Laboratory, Sacramento, CA samples were very low (1-10 copies) indicating that the virus was shed at a low level, which might explain that only 50% of the samples

687 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

(4 out of 8) were found positive. In summary, this study is the first sensors changed from low to low-medium. A low reading indicates evidence of the detection of a major zoonotic agent such as LCMV by ammonia levels of 0-1 ppm. The second major color change occurred exhaust air dust in IVC racks. This alternative health monitoring on day 14 where the levels changed to medium. A medium reading technique may consequently be considered for the detection of indicates ammonia levels of 1-25 ppm. The third color change prevalent and zoonotic agents. occurred on day 19 where the levels changes from medium to medium-high. A high reading indicates ammonia levels of 25-50 P303 Dermal and Subcutaneous Mapping Assessment of Yorkshire ppm. Color-metric interpretation of the intracage ammonia levels Swine via Ultrasound revealed that the maximum exposure did not extend past 25 ppm and was considered within acceptable range. The company who S Gerth*, W Woodley, B Selvage, R Pettis, N Bolick produces the sensors were consulted to confirm the interpreted results. Future investigations will focus on analysis of the effects of Parenteral Sciences, BD Technologies, Durham, NC prolonged ammonia exposure limits through histological samples of nasal epithelial tissue. Swine are a common injection and device research model as their subcutaneous and dermal tissue structure are highly representative P305 Comparison of 3 Formulations of Buprenorphine in C57BL/6 of published human dermis and subcutis mean thicknesses: arm Mice (dermis 2mm, SD 0.4/subcutis 11mm, SD 5.6), abdomen (2mm, SD 0.4/14mm, SD 7.3), buttock (2mm, SD 0.5/15mm, SD 7.3), and thigh A Dickerson*1, P Myers1, D Goulding1, R Wiltshire2, K Laber2, F Lih4, (2mm, SD 0.4/10mm, SD 5.7)1. Literature cites the swine neck, M Comins1, CA McGee3, TL Blankenship-Paris1 inguinal, and flank anatomy as sites for injection assessment. This study uses ultrasonography of the neck, inguinal, and flank of 1Veterinary Medicine Section, NIEHS/NIH, Durham, NC; 2Office of domestic Yorkshire farm swine (n=6) to define thickness of dermal the Chief, NIEHS/NIH, Durham, NC; 3Clinical Research Unit, and subcutaneous tissue strata as a function of age and weight. NIEHS/NIH, Durham, NC; 4Epigenetics & Stem Cell Biology Measurements taken at monthly intervals from age 10 wk (~24kg) to Laboratory, NIEHS/NIH, Durham, NC 30 wk(~80kg) will serve as a guide in matching swine anatomy and age with target tissue morphology. Predetermined sites and surface Buprenorphine is a commonly used analgesic in rodent surgery; area per location were assigned at study start and followed however, dosing must be repeated every 12 h for adequate analgesia. throughout: right, left, and back neck (25cm2each), right and left This redosing requires repeated handling postsurgery causing added inguinal (32cm2 each), right and left flank (147cm2 each). Sonograms stress. Sustained-release buprenorphine, which reportedly maintains from sites were used to measure dermal (skin top to dermis bottom) blood levels of buprenorphine greater than 1 ng/ml (reported and subcutis (skin top to subcutis bottom) depths. Subcutis thickness therapeutic concentration levels) out to 48-72 h is commercially was determined by subtraction of dermis from subcutis depth. In available. The viscosity of the product with small dosing volumes 10-wk-old swine (~24kg), subcutis tissue thickness trends as follows: make accurate dosing a challenge. A concentrated formulation of mean flank (10mm, SD 1.2) > neck and inguinal (8mm, SD 2.7 and buprenorphine HCl, labeled for use in cats with recommended 1.9); in dermis, mean neck (4mm, SD 0.43) > inguinal and flank dosing every 24 h is commercially available. We evaluated this (2mm, SD 0.45 and 0.18). Subcutis thickness at 14 wk (~32kg): neck product in male and female C57BL/6NCrl mice. We examined serum (11mm, SD 3.3) > flank (10mm, SD 2.1) > inguinal (8mm, SD 1.4); in concentrations over time following a single injection and compared dermis, neck (4mm, SD 0.58) > inguinal and flank (2mm, SD 0.19 and to standard buprenorphine (BUP), a high concentration 0.22). Among the 3 locations, neck has the thickest dermis (~4mm), buprenorphine formulation (SIM), and the sustained-release product twice of inguinal or flank (2mm) and literature for typical human (BupSR). Males and females were injected with 1 of the 3 dermis (2mm). The flank has the largest surface area (294cm2 total) buprenorphine formulations at a dose of 1mg/kg subcutaneously at allowing for higher n value studies requiring fewer swine. The time 0. Groups of mice (n=8) were sacrificed at 1, 4, 8, 12, 16 h for all Yorkshire swine neck, inguinal, and flank were all found feasible for groups and 24 h for the SIM and BupSR groups. Blood was collected, subcutaneous and dermal injection studies dependent on specific serum separated, and liquid chromatography-mass spectrometry study requirements for depth and surface area. Continuation of (LC-MS) used to determine concentrations of buprenorphine in each measurements to higher weight and age ranges remains ongoing and serum sample. High blood level concentrations were observed in may further delineate sites. both SIM and BUP groups 1 h following injection (56 ng/ml and 51 ng/ml, respectively). Both groups had similar pharmacokinetic P304 A Novel Intracage Ammonia Sensor curves including rates of decline. BUP groups had mean concentrations <1ng/ml by 12 h and the SIM by 16 h. The BSR T James* concentration was 5.3 ng/ml and remained above the 1 ng/ml therapeutic threshold throughout the 24 h. We conclude that SIM University of South Florida, Tampa, FL does not offer sustained-release properties and does not offer any dosing advantages over BUP. A behavioral study requires prolonged cage changes due to potential disruption of the animal’s circadian rhythm. Prolonged cage P306 Comparing 4 Cryopreservation Methods for Histology and changing raises concerns of increased ammonia levels which may RNA Extraction affect the animal’s health. The principle investigator requested changeouts on a 3-wk cycle. A pilot experiment was conducted to V Nunez*1, A Cook1, C Havnar1, S Flanagan1, M Lu2, RE Taylor1, A examine the impact of cage change frequency for singly housed mice Martzall1, O Foreman1 on corn cob bedding within controlled environmental units. To investigate whether that length of time between cage changes was 1Pathology, Genentech, Inc, South San Francisco, CA; 2Veterinary acceptable, ammonia levels were measured over a 21-d period. Sciences, Bristol-Myers Squibb, Redwood City, CA Currently, it is unclear what the exact maximum exposure of ammonia is for mice, however for humans the OSHA regulatory Frozen sections are usually used where there is a need to preserve limits are currently at 50 ppm and the NIOSH recommended limits nucleic acids or when the antigen being visualized by IHC is are at 25 ppm. Eight small animal ammonia sensors were placed sensitive to crosslinking fixation techniques. Cryosectioning requires inside cages within 2 units. Notation of color changes indicating the freezing of fresh tissue in optimal cutting temperature compound shifts in ammonia levels were recorded daily as well as room (OCT). Traditionally, chilled 2-methylbutane is used as the freezing temperature/humidity and animal health status. Pictures of the reagent; however, it is toxic, extremely volatile, and flammable at sensors were taken on various days throughout the 21-d period. The room temperature, and requires special handling and safety first color change occurred on day 7 where ammonia levels for all 8 procedures. The goal of this study was to test the performance of 4

688 Abstracts of scientific papers 2019 AALAS National Meeting

cryopreservation protocols and identify suitable and safer Swine are frequently used as models for medical device and alternatives to 2-methylbutane. Three protocols used commercially procedure testing due to their comparability to humans in size and available freezing reagents and the fourth consisted of snap-freezing vascularization. Qualitative comparisons illustrate the similarity the tissues in liquid nitrogen. The freezing methods were compared between swine and human cardiovascular systems, but quantitative on the following criteria: ability to preserve microscopic appearance, comparisons are not readily documented. The purpose of the study ability to preserve RNA integrity, and assess overall protocol was to measure blood velocity and vessel diameter in commonly duration, complexity, and safety. All major organs were harvested accessed swine peripheral vessels and make quantitative from 3 adult female CD-1 mice. Tissues were divided into 4 portions, comparisons with previously published data on blood flow in human placed in an OCT filled cryomold, and frozen by 2-methylbutane, blood vessels. Thirteen male Yorkshire swine, weighing at least 60kg, 1-methoxyheptafluoropropane, hexane, and 1 portion was frozen were used in this study. The swine were anesthetized and directly in cryotubes placed in liquid nitrogen. Purified RNA samples ultrasonography was used to visualize peripheral vessels. Vessel were extracted and a full-spectrum, UV-Vis spectrophotometers was diameter and systolic velocity measurements using pulsed wave used to quantify and assess purity of RNA. The RNA integrity (PW) doppler were taken for multiple vessels on each swine subject. number was determined using an automated electrophoresis tool for Ten vessels were visualized and measured for each animal: the right RNA sample quality control. For most tissues, using the saphenous artery, right cephalic artery, right femoral artery, right 1-methoxyheptafluoropropane freezing protocol provided excellent saphenous vein, right cephalic vein, left saphenous artery, left preservation of microscopic architecture and RNA quality that was cephalic artery, left femoral artery, left saphenous vein, and the left not significantly different from samples frozen using 2-methylbutane cephalic vein. Vessel diameter and maximum flow rate velocity were or hexane methods. Flash freezing in liquid nitrogen remains the averaged across all animals. Comparisons were made to published gold standard for RNA preservation and produces the highest quality scientific studies on blood flow in human blood vessels within the RNA; however, when histology is required in addition to RNA forearm, abdomen, chest, and leg anatomical regions of the body. extraction, the 1-methoxyheptafluoropropane method should be Mean values of the peak systolic velocities in the left and right swine considered as a suitable compromise. In addition, the saphenous (μ = 317.5 mm/s, σ = 116.1 mm/s) and femoral artery (μ = 1-methoxyheptafluoropropane is less toxic and flammable than 645.03 mm/s, σ = 168.23 mm/s) had overlapping standard deviations Hexane and 2-methylbutane, and offers a safer method for freezing with the systolic velocity in the human radial (μ = 313.8 mm/s, σ = tissues. 72.2 mm/s) and ulnar (μ = 545.5 mm/s, σ = 104.5 mm/s) arteries from previous studies. Additionally, swine vessels showed no P307 Pharmacokinetics and Pharmacodynamics of Maropitant, differences when comparing the left and right sides. Additional Alone or in Combination with Buprenorphine, Using an Incisional vessels have shown similarities and differences between swine and Pain Model in Rabbits human data. This provides qualitative evidence for the use of swine as a translational model for procedure and medical device W McGee*1, G Au2, LV Kendall1, D Gustafson3, P Boscan3, M Sadar3 development and represents a refinement of the swine vascular model with potential for reduction of animals. 1Laboratory Animal Resources, Colorado State University, Fort Collins, CO; 2College of Veterinary Medicine, Washington State P309 Development of Vascular Endothelial Growth Factor-induced University, Pullman, WA; 3Clinical Sciences, Colorado State Retinal Vascular Leakage Model in Rabbits University, Fort Collins, CO X Ping*1, L Yang2, J Destefano1, C Nunes1, C Li1, K Lodge1, L Rabbits experience difficult recoveries after anesthesia due to Gichuru1, W Dong1, D Liu1, T Gray1, C Johnson1, S Motzel1, N Li2, X decreased appetite and gastrointestinal stasis. Analgesic side effects Shen1 include gastrointestinal ulceration with NSAIDs and gastrointestinal stasis with opioids. This study used an incisional pain model to 1SALAR, Merck, West Point, PA; 2Cardio-Metabolic, Merck, West determine the effects of maropitant and buprenorphine on pain, Point, PA appetite, and fecal output in rabbits. Twenty female New Zealand white rabbits were divided into 5 groups: 1) buprenorphine 0.06mg/ Development of novel drugs and drug delivery mechanisms, as well kg IM q8h, 2) buprenorphine+low-dose maropitant 5mg/kg SC q24h, as advanced ophthalmological techniques, requires experimental 3) buprenorphine+high-dose maropitant 10mg/kg SC q24h, 4) high- models with animals capable of developing ocular diseases with dose maropitant, or 5) saline. Rabbits were anesthetized with similar etiology and pathology and suitable for future studies of new isoflurane and treatments started at anesthesia onset. An incision was therapeutic approaches. Although experimental ophthalmology created on the plantar surface of the right pelvic foot, and tactile research is traditionally performed on rodent models, these animals sensitivity measured using a von Frey device at baseline and 3, 6, 12, are often unsuitable for preclinical drug efficacy studies or testing 18, 24, and 48 hours postoperatively. Food intake, weight, and fecal novel drug delivery approaches. Therefore, rabbit models of ocular output were measured daily. Three rabbits received maropitant 10 diseases are particularly useful, since rabbits can be easily handled mg/kg SC for pharmacokinetic sampling. Samples were taken 0, 0.5, and share more common anatomical and biochemical features with 1, 2, 4, 6, 12, and 24 hours after drug administration. Some rabbits humans compared to rodents, such as longer life spans and larger tolerated less pressure with von Frey testing after surgery, but there eye size. Diabetic retinopathy (DR) and diabetic macular edema were no group differences in tactile sensitivity. Rabbits from (DME) are characterized by a series of retinal microvascular changes buprenorphine-treated groups ate less and had reduced fecal output and increases in retinal vascular permeability. To develop an after surgery compared to those that did not receive buprenorphine. appropriate PK/PD model to define pathophysiological progress of The maximum plasma concentration of maropitant occurred at 30-60 retina vasculature, a vascular endothelial growth factor (VEGF) min with a half-life of 12.6±2.3 h and a clearance of 6.74±0.17 L/h/ induced retinal leakage model was established in rabbits. kg. While buprenorphine decreased food intake and fecal output, Noninvasive scanning ocular fluorophotometry was chosen to maropitant did not have an effect on pain, food intake, or weight at measure free sodium fluorescein leakage from the retinal and iris the doses and frequency used in this study. vasculature. Single Intravitreal injection (IVT) of VEGF induced a dose-dependent breakdown of blood-retinal barriers with maximal P308 Swine as a Translational Model: Qualitative Comparisons of leakage level at 48-72 h postinjection. VEGF-induced leakage was Vessel Diameter and Systolic Blood Velocity in Swine Peripheral completely blocked by anti-VEGF (ranibizumab) and 2 steroids Blood Vessels to Humans Blood Vessels (triamcinolone acetonide (TAA) or dexamethasone (DEX) which are standards of care (SOC) for DME therapies, through either single WA Spinks*, C Baschiano, CZ Cannon, S Gabriel intravitreal injection or systemically administration for 3 d. As a result, this post demonstrated that a VEGF-induced retina vascular BD Technologies, Durham, NC leakage model in rabbit can be used for DME research.

689 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

P310 Differential Blood Buprenorphine Levels in Mice Treated showed mitochondrial appearance and intra-mitochondrial cristae with Sustained-release Buprenorphine Pre- Versus Post-Isoflurane network, respectively. Treatment of sperm with hydrogen peroxide Anesthesia induced a decrease in mitochondrial membrane potential. The effects

of H2-treatment on mitochondria in damaged sperm are under YS Moreno*, J Stathopoulos, T Caron investigation. Because of the rapid diffusion and high membrane

permeability, H2 can reach and react with intrasperm ROS, including Comparative Medicine, The Broad Institute, Cambridge, MA hydroxyl radical, possibly in mitochondria and improve low sperm

motility. The results strongly suggest that H2 is a new promising tool Sustained-release buprenorphine (BUP-SR) is widely used as a for male infertility treatment. postoperative analgesic in mice, where a single dose is reported to provide sustained analgesia for up to 72 h. To achieve this duration P312 Genetic Loci for Resistance to Podocyte Injury Caused by the of action, the BUP-SR formulation relies on a liquid polymer matrix Tensin2 Gene Deficiency in Mice that acts as a subcutaneous depot, controlling the rate of drug release into the bloodstream. Recently, we observed behaviors suggestive of Y Takahashi*1, H Sasaki1, K Hiura1, K Nakano2, T Okamura2, N opioid overdose in mice treated with appropriate doses of BUP-SR Sasaki1 prior to isoflurane anesthesia. Such behaviors were absent in animals treated with an equivalent dose of BUP-SR subsequent to isoflurane 1Kitasato University, Towada, Japan; 2Research Institute National anesthesia. We hypothesized that these behavioral differences may Center for Global Health and Medicine, Shinjyuku, Japan have been the result of different rates of buprenorphine release due to compromised integrity of the liquid polymer matrix after exposure The ICGN mouse is a model of glomerular dysfunction that shows to isoflurane. To determine the effect of isoflurane exposure on blood gross morphologic changes in the podocyte foot process levels of buprenorphine in BUP-SR treated mice, 2 groups of 5 female accompanying proteinuria. Previously, we demonstrated that C57BL/6 mice were given a single subcutaneous injection of BUP-SR proteinuria in ICGN mice is caused by the deletion mutation in just prior to, or just after 1 h of isoflurane anesthesia. Blood samples the Tensin2(Tns2) gene.Tns2 is a focal adhesion-localized were collected at predetermined intervals until 48 h post BUP-SR multidomain protein expressed in various tissues, and its administration and buprenorphine levels determined via ELISA dysfunction leads to alterations in podocytes. To identify the modifier assay. Mice were also observed for clinical signs associated with gene(s) to glomerular dysfunction, we produced congenic strains opioid overdose for 180 min subsequent to BUP-SR dosing. Mice carrying the Tns2-null mutation (nph) on the several genetic treated with BUP-SR before isoflurane anesthesia achieved relatively backgrounds and analyzed their severity. Interestingly, the C57BL/6J steady blood concentrations of buprenorphine, peaking at 0.723 ng/ (B6) and 129/SvJcl mice congenic mice exhibited milder phenotypes mL 180 min post BUP-SR administration and maintaining a steady than did ICGN, DBA/2, and FVB strains.Thus, we performed a blood level for up to 48 h post dosing. Conversely, blood genome-wide linkage analysis of backcrosses between two Tns2- concentrations of mice treated with BUP-SR after isoflurane exposure deficient mouse strains, B6. ICGN-Tns2nph(resistant) and FVB. ICGN- rose to 1.4 ng/mL at 180 min post administration (P = 0.0003), Tns2nph(susceptible), and detected a novel major modifier locus on remained above 1.2ng/mL for 24 h (P = 0.01), then fell to levels chromosome 10. The combined effect of the C57BL/6J alleles of the 2 comparable with animals dosed before isoflurane exposure at 48 h. loci on chromosomes 2 and 10 reduced the urinary albumin excretion Behavioral indices did not differ between groups during the caused by Tns2 dysfunction to a level comparable to that of observation period. While these findings do not necessarily explain C57BL/6J mice. These data indicate that the resistance to podocyte our initial observations, it does appear that the timing of BUP-SR injury caused by Tns2 dysfunction is mainly produced by the effects treatment relative to the administration of isoflurane anesthesia may of the modifier genes on the two loci. The identification of these lead to profound differences in blood buprenorphine levels in mice, modifier genes is expected to help elucidate the mechanism potentially affecting the adequacy and duration of analgesia. underlying podocyte injury.

P311 Protective Effect of Molecular Hydrogen on Oxidative Stress- Platform Sessions induced Impairment of Mouse Sperm Motility PS1 Is it Time for the Certified Lab Animal Diet Contaminant Y Noda*1, T Nemoto1, T Endo1, I Ohsawa2 Standards to be Updated?

1Animal Facility, Tokyo Metoropolitan Institute of Gerontology, D Barnard* Itabashi-ku, Japan; 2Biological Process of Aging, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, , Japan Office of Research Sources/Division of Veterinary Resources, National Institutes of Health, Bethesda, MD To investigate the effects of H2 on damaged sperm, suspension of fresh sperm obtained from B6D2F1/Crlj mice (12-15 wk of age, 3 The concept of certified lab animal diet was developed during the males) was treated with 0.3 mM hydrogen peroxide for 30 min, and late 1970s. Certified lab animal diets guarantee that the further incubated with or without H2 for 20 min. To evaluate concentrations for 5 heavy metals, aflatoxin, polychlorinated fertilizability, H2-treated damaged sperm (2 males) was used for in biphenyls (PCBs), and 23 pesticides do not exceed set limits. It is the vitro fertilization (13 females). The obtained 2-cell stage embryos recommended diet for good laboratory practices (GLP) studies. As were transferred to pseudopregnant Jcl:MCH (ICR) mice (3 females). part of the NIH lab animal diet quality assurance program, heavy To further investigate the effect of H2 on ROS-dependent metal and pesticide analyses have been performed on 15 lab animal mitochondrial damage, we stained sperm with NAO, mitochondrial diets for 25 years. The QA pesticide screen included 22 of the 23 cardiolipin binding dye, and MitoTracker Red, mitochondrial pesticides on the certified diet list. Over the 25 years of analyses only membrane potential-dependent dye, and observed them with STED malathion, on occasion, exceeded its minimum detectable super-resolution microscope. The structure of sperms was observed concentration (.01 ppm). Since 1990 14 of the pesticides listed for by SEM. Fresh sperm (motility rate: 82.4%) were treated with certified diets have been banned from use. The Environmental hydrogen peroxide, resulting in damaged sperm with low motility Protection Agency has reported on the 25 most used pesticides in rate (14.6%). H2-treatment significantly restored their motility rate agriculture, and none of these pesticides are on the certified diet (63.9%) accompanied by improvement of intrasperm ATP content. pesticide list. Analyses for glyphosate, the most used agricultural Fertilization rate of damaged sperm was markedly improved from pesticide, in 3 lab rodent diets from 3 lab animal diet manufacturers 37.6% to 59.2% by H2-treatment. Transfer of 2-cell stage embryos showed the average concentration was 0.94+ 0.178 ppm. During the obtained from H2-treated damaged sperm showed normal ontogeny last 19 y mycotoxins and isoflavones have been included in the NIH (94.6%). SEM imaging and STED imaging of NAO-stained sperm lab animal diet QA analyses. Aflatoxin the only mycotoxin on the

690 Abstracts of scientific papers 2019 AALAS National Meeting

certified diet contaminant standards list has not been found in any individuals exhibiting FS (n=7). Observations were made for the diets. However, vomitoxin, fumonisins, ochratoxin A, and following experimental periods: baseline, two repeated trials (with zearalenone are found in the diets. It appears that the contaminant paint), and post-trial (without paint). Finger paint was distributed in standards for lab animal certified diets need to be reviewed and plastic containers secured to caging daily after husbandry during the updated. This will require removing contaminants that are no longer 2 repeated trials to evaluate the effectiveness of this intervention. a hazard, determining new contaminants to add to the certified diet Over the course of this project, a trend towards lower FS scores was standards, and, determining the maximum acceptable levels for the observed for day 14 for individuals utilizing finger paints. There was new contaminants. a significant reduction (P = 0.026) of FS between baseline and finger paint trials on day 14. As a result of this evaluation, the use of finger PS2 Using Clickers to Quiet Monkey Chaos: An Overview of paints in response to FS has been incorporated in conjunction with Implementing a Husbandry-based Clicker Training Program destructible enrichment as part of the standard intervention response. AJ Schenk* PS4 Continued Evaluation of Extended Cage Change Interval for Division of Laboratory Animal Resources, Duke University, Durham, Breeding Mice NC M Dickerson*, R Van Andel Nonhuman primate rooms can be a hectic and noisy environment for all involved. Calming and shaping that behavior can be challenging University of Utah, Salt Lake City, UT in large nonhuman primate colonies due to limited staffing and time constraints. However, literature review reveals that animals engaged The optimal cage change interval for laboratory mice remains in positive reinforcement training display less stress, aggression, and controversial. Our institution previously used a 16-wk cage change stereotypical behaviors due to the sense of control, human interval for mice. This study was undertaken to evaluate whether interaction, and the psychological enrichment the animals recieve cage sanitization interval affects breeding performance, cage from these interactions. Previous studies in canine shelters have microenvironment, and animal health and behavior of mice housed shown that clicker training shelter dogs led to decreased barking and in individually ventilated cages (IVC) with pelleted paper bedding. an increase in positive behavior changes. Applying the framework Continuous and noncontinuous breeding trios of C57BL/6 mice were from these studies, we modified the daily husbandry routine for our followed longitudinally over 18 wk. Cages were completely changed nonhuman primate staff. We incorporated key aspects of using the every 2 wk or had a partial bedding change (75%) with a complete clicker when providing food enrichment items to animals that change at 16 wk. A total of 20 cages were followed, 5 per study displayed positive behavior traits. Animals that demonstrated condition. Breeding success was determined by analyzing litter size, aggressive behavior were not provided extra food enrichment items pup weights, and litter mortality. We evaluated cage and the clicker was not used. Additionally, this interaction helped microenvironment by assessing microbial load and air quality, and develop the bridge of the clicker for more advanced behavior we subjectively scored the cage condition, animal appearance, and training. Implementation of this program was simple and well animal behavior every 2 wk before and after cage change or partial received by the husbandry staff. Staff members received training bedding change in both continuous and noncontinuous breeding from the nonhuman primate enrichment and behavior group on how groups. We found no significant difference in breeding efficiency to use the clicker and recognize positive versus negative behavior between groups. Although there were no differences in animal traits. Initial subjective feedback from the husbandry, veterinary, and behavior between groups, we did note improved animal appearance enrichment staff reported favorable results. Data generated from a in the partial cage change groups that was statistically significant for comparison of the behaviors displayed by these primates before and the continuous breeding trios. Ammonia levels were significantly after the initiation of the clicker training program will be presented. higher in the partial change cages. Cage wall bacterial levels were This work tests the value of clicker training for inducing positive higher in the extended sanitation group and taxonomic differences behavioral changes in non-human primates and explores were seen between the groups using microbiome analysis. Extended methodologies and metrics for implementing a husbandry based cage change did not improve breeding outcomes or animal welfare nonhuman primate training program. based on behavioral analysis. Cage air quality and microbial growth were worse with extended cage change and the practice may alter the PS3 Finger Painting: A Novel Intervention for Feces Smearing in microbiome. Our data do not justify an extended sanitation interval Nonhuman Primates for breeding mice housed on pelleted paper bedding.

C Osimanti*, W Chan, M Hogan PS5 Is a Once per Week Cage Change Possible with Diabetic Mice?

Center for Comparative Medicine, Massachusetts General Hospital, K Nacel*1, J Delatorre1, L Kramer2 Charlestown, MA 1AST, AstraZeneca, Gaithersburg, MD; 2Lenderking Caging Products, As a component of the environmental enrichment program, behavior Millersville, MD management includes observation, quantification, and intervention for abnormal or undesirable behaviors in laboratory nonhuman Studies involving diabetic mice are challenging since they are primates (NHPs). Feces smearing (FS) is a stereotypic behavior sensitive to distress that can affect the onset of diabetes. Polyuria observed in approximately 5.88% (6/102 NHPs) of rhesus (Macaca causes cages to soil quickly and remain wet, which means near daily mulatta) and cynomolgus macaques (Macaca fascicularis), and baboons cage changes. Such frequent cage changing can induce stress in mice (Papio anubis) at this institution. Currently, standard institutional and increase expenses of labor and consumables, such as food and interventions provided to the NHPs designed to curb FS include bedding. A novel cage design that uses a perforated false bottom to destructible enrichment (e.g. box filled with paper and food items), hold bedding while filtered air is pushed through the bedding was with the addition of therapeutic enrichment (e.g. foraging board), compared with a traditional IVC system housing 5 diabetic (DbDb) expanded cage space, or a water cup filled with food-grade baking mice per cage. The traditional IVC was changed 3 times per week, extract as needed. Several studies and case reports on autistic and the novel IVC system changed once per week. Metrics of children who exhibit neurological and behavioral limitations have ammonia, temperature, relative humidity, and moisture content of shown the potential use for finger paints as a treatment for these the bedding were collected in both caging systems. Ammonia, dysfunctions. We hypothesized that the use of finger paints as a temperature, and relative humidity were collected using wireless novel intervention would lead to a reduction of FS in NHPs. sensors which collect data without opening the cage. Moisture Observations were made 6 h, 24 h, 7 d, and 14 d after cage change for content was measured at 4 corners and the center of the cage.

691 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

Traditional cages displayed low ammonia throughout the cage- NC3Rs 2010 study discussing moving mice from dirty to clean cages change cycle, whereas the novel cages saw ammonia increase on day by using tunnels or hands. The study showed that using hands or 5 but remain under 25 ppm by day 7 of the cage change. We could tunnels was less aversive for mice than using forceps, and resulted in not allow the traditional cage to go out 7 d due to the wetness, and fewer stress behaviors observed after cage change. Following this we had to use a 2-3 days cage change cycle during this comparison experience, in May 2017, we obtained both IACUC and principal study. Both cages stayed within the preferred temperature range. investigator approval to implement tunnels as part of the cage Relative humidity was higher in the novel cage by the end of the 7 change process for our immunocompromised mice. We made no days as compared to the traditional cage at 3 days. Moisture content other changes to the husbandry of the room, staff, nor to the measurements found more dry corners and overall lower total cage researchers using the room. We continued making clinical average in the novel cage as compared with the traditional cage. Our observations looking in particular for additional skin lesions. An study indicates that the novel caging system would extend the cage analysis of cases from May through November 2017 showed a 100% change interval for diabetic mice. Fewer cage changes will reduce reduction in skin lesion cases. A more recent check in May 2019 labor and supply costs. Reducing the cage changing requirements showed that no further skin lesions have been observed in the NSG may also aid in reducing the stress associated with handling. The mice. Due to the success in implementation of tunnels in our goal of this study is to safely extend the cage change time period for immunocompromised room, we’ve expanded to make this our these diabetic models in the novel cage. standard model for mouse husbandry across the facility.

PS6 Temperature and Relative Humidity in Static and Ventilated PS9 Going beyond the Isolator: A Novel Process and Facility Cages Housing Mice (Mus musculus) Design to Acknowledge the Role of the Macroenvironment In Germfree Rodent Derivation Success TJ Poling*, N La Santa Medina, DS Crowell, IM Washington C Weiner*, PL Roesch Office of Laboratory Animal Resources, West Virginia University, Morgantown, WV Taconic Biosciences, Inc., Rensselaer, NY

The Guide recommends 68-79F temperature (T) and 30-70% relative As the field of germfree and gnotobiotic rodent science grows to humidity (RH) for mouse macroenvironment. Intracage T and RH support microbiome and gut flora-specific inquiries in basic and may be affected by factors including macroenvironment, cage type, advanced research, the need to generate strain-specific animals at animal number, activity, bedding, and cage change frequency. Our germ-ree health status in vivo also arises. This derivation process is objective was to measure intracage T and RH and evaluate potential technically difficult when limited to working within the modulating factors. We hypothesized that T and RH are higher in microenvironment of the flexible film isolators, as the physical and static versus individually ventilated (IVC) cages and vary with mechanical constraints of a bioexclusionary containment structure macroenvironment, animal number, diurnal cycle, and bedding type. prevent the efficient and successful outcome for the many Male or female mice were housed at 1, 3, or 5 per cage in static or IVC coordinated parts of the biological derivation process. Constraints cages in 2 rooms maintained at 12h:12h light:dark cycles. Data include the lengthy contact time of chemical sterilants used in port loggers in cage lids, rooms, and outside the building provided hourly entry and closure; the use of anesthesia, analgesia, and surgical T and RH readings for 28d. Corncob or bedding consisting of equipment within an isolator, and limitations of cylinder volume for specially processed hardwood paper pulpwere each used for 14d, needed materials. Further, the historical approach to germfree with cage changes at 7d (static) or 14d (IVC). T and RH readings isolator work relies on large quantities of toxic chemicals to clean up were compared by T-test or ANOVA using significance at P < 0.05. the macroenvironment and facilitate the success of isolator port entry Bedding type and cage change frequency did not affect T or RH in of materials and supplies. Many existing facilities are not designed to any cages. T was higher in IVC vs static cages housing 1, 3, or 5 mice work with these materials, processes, and equipment successfully, but did not vary directly with mouse number. T was higher thereby limiting the potential broad-reaching scope of gnotobiotics. nocturnally in static and IVC cages housing 3 or 5 mice. RH was As the microbiome field continues to expand, the need to manipulate higher in static vs IVC cages housing 1, 3, or 5 mice and increased and handle these germfree animals outside the physical constraints with mouse number in static cages. No diurnal RH variation of the isolator grows, as does the need for ventilation support for occurred in any cage. Intracage T and RH were both related to room chemical use within animal facilities. Therefore, an understanding of parameters. In summary, our findings included lower T and higher the baseline and ongoing cleanliness of the macroenvironment must RH in static vs IVC cages, diurnal variation in T but not RH in static be considered. The process for conducting germfree derivations is and IVC cages, and RH but not T variation with mouse number in reframed around working in a sterile macro and microenvironment. static cages. Thus, microenvironmental T and RH are affected by We hypothesize that the clean design of the macroenvironment will several factors and should be considered for relevance to animal improve germfree derivation success rates as well as mitigate and/or welfare. dramatically reduce the need for harsh chemical sterilants. The process for conducting a germfree derivation and embryo transfer in PS8 Mouse Handling with Tunnels a biosafety cabinet is described. In order to maintain acceptable success rates, which is measured by germfree derived pups, we’ve K Marshall*, H Wolford, K Kimura, L Martin developed a novel decontamination process to mitigate potential contamination scenarios based on identified high-to-low risk points Oregon National Primate Research Center, Oregon Health and in the derivation process. Also, we’ve developed a novel facility with Science University West Campus, Beaverton, OR higher standards of macroenvironmental engineering and equipment controls compared to standard animal and surgical facilities, and we Our facility houses 1,000 mice. Approximately 20% of our census are describe the process that drove us toward this concept and outline immunocompromised, including NSG™ mice, a severely our parameters for success. immunodeficient model generally unable to fight off infections. In 2016, we observed a significant increase in skin lesion reported in this PS10 The Impact of Visual Cues on Rabbit Behavior population; 19 cases reported in a 6-mo window versus our typical annual caseload of 1-2 cases. Two factors potentially contributing to NA Castrejon*, ML Martin, CC Hofer the increase included a new staff member and the introduction of Staphylococcus xylosus. This challenge provided our team with a Small Animal Medicine, Walter Reed Army Institute of Research, new opportunity. Recently, the Unit Manager traveled overseas on an Silver Spring, MD AAALAC International Fellowship Award, where she visited various facilities in and around London. There, she was introduced to an Rabbits (Oryctolagus cuniculus) are known for being curious and

692 Abstracts of scientific papers 2019 AALAS National Meeting

timid animals, are easily frightened, and can be difficult to manage in A 1-y-old, castrated, pair-housed male ferret was examined for a laboratory setting. As lab animal technicians we are bound by the lethargy, weight loss, and pale mucous membranes. The animal was Animal Welfare Act, the Guide, the IACUC, and our own personal used solely for annual serology for influenza screening of the naïve morality to mitigate any unnecessary stress to the animals under our ferret colony. On physical examination, the animal had appropriate care. In an effort to alleviate stress during acclimation, handling, and mentation, ideal body condition (3/5), pale mucous membranes, no husbandry duties, visual cues were initiated in order to indicate to murmurs or arrhythmias, normal lung sounds, and no palpable the rabbits what activity was about to occur. After the acclimation abdominal masses or fluid. The animal was anesthetized with 1-5% period and at the start of the protocol, the rabbits were exposed to isoflurane in an induction chamber before being transferred to the laminated signage which hung on the wall facing all caging and working table on a tightly fitting facemask and maintained on 1-3% remained visible to animals at all times. The team used signs isoflurane for a jugular blood collection. A comprehensive analysis consisting of 1 colored shape designated for each procedure type: including a complete blood count, serum chemistry, packed cell husbandry/enrichment (blue circle), invasive procedures (red volume, total protein, and blood smear was conducted. The triangle), or status quo (green square). The signs were placed no less comprehensive analysis revealed an elevated creatinine (0.8mg/dL), than 1 h prior to any activity. This allowed the animals’ time to severe non-regenerative, normocytic, normochromic anemia (PCV observe the sign change and prepare for human interaction. 6%); all other values were within normal limits. Blood smear revealed Behavioral changes were measured objectively by technicians on a a lack of reticulocytosis. The differential diagnosis included pure red weekly basis. Within 2 wk of the sign placement significant changes cell aplasia, chronic kidney disease, immune-mediated anemia, and in behavior were noted. The rabbits tended to come to the front of the neoplasia. The animal was prescribed erythropoietin (150 IU/kg IM), cages and appeared to be more compliant in our presence. There was iron dextran (10mg/kg IM), ad lib carnivore care (6.4 tbsp dry/kg a marked reduction in foot “stomping” or thumping and hiding, and PO) and daily polyvit (0.5mL PO) for 4 d. The anemia was less resistance when attempting to pick them up. The value of unresponsive to therapy after three days and, due to animal welfare communication is often overlooked when dealing with small concerns as well as study-related contraindications of prolonged laboratory animals but the potential benefits are not to be immunomodulation therapy, euthanasia was elected. Gross necropsy underestimated. The change in behavior has been paramount and revealed general mucosal pallor and mesenteric lymphadenopathy. reflects in the success of our study and our relationship with our Bone marrow histopathology revealed hypercellular marrow with an rabbit population. increased myeloid to erythroid ratio due to selective erythroid hypoplasia. These findings could be compatible with pure red cell aplasia, which is only very rarely described in ferrets. Successful PS11 Mass in a Northern Tree Shrew (Tupaia belangeri) management of pure red blood cell aplasia involves prolonged therapy with multiple immunomodulating drugs (prednisone, SL Spears*2, AK Brice2,1, L Makaron1 cyclosporine, and azathioprine).

1University Laboratory Animal Resources, University of PS13 Respiratory Complications in a Rhesus Macaque (Macaca Pennsylvania, Philadelphia, PA; 2Department of Pathobiology, mulatta) with Chronic Indwelling Catheter University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA V Mrotz*1,2

A 2-year-old, intact female tree shrew (Tupaia belangeri) presented for 1Center for Comparative Research , University of Mississippi Medical recurrent mild periocular dermatitis. As an incidental finding during Center, Jackson, MS; 2Center for Diseases Control and Prevention, physical examination, a fluctuant, subcutaneous, 1.5cm x 1.0cm mass Atlanta, GA at her second left abdominal mamma, centered underneath the nipple was identified. Lymph nodes were unremarkable. Very little is A 10-y-old, singly housed, male Rhesus macaque (Macaca mulatta), published on mammary masses of tree shrews though differentials involved in a chemical dependency protocol that uses chronic from other laboratory animal species include intraductal tumor, indwelling catheters for self-administration, was examined for acute mammary adenoma/adenocarcinoma, mastitis, myoepithelioma, and inappetence and changes in study responses. Cage side assessment lymphoma. A fine needle aspirate was taken of the mass revealing a found the macaque to be bright, alert, responsive, and the indwelling uniform population of neoplastic epithelial cells. Surgery was right internal jugular catheter was flushing appropriately. Under scheduled for a planned excisional biopsy. Two weeks later at the ketamine (10 mg/kg, IV) sedation the macaque was notably dyspneic time of surgery, the mass had grown another 30% to 1.9cm x 1.2cm x with pale blue mucus membranes. Supplemental oxygen was 0.9cm. The tree shrew was induced with isoflurane (2% isoflurane in provided via facemask for the remainder of the physical exam, which 100% oxygen). Full body radiographs did not show any evidence of highlighted crackles auscultated over the chest, predominately on the metastasis. Preemptive analgesia of buprenorphine sustained release left side, and a small amount of purulent discharge at the catheter (0.25mg, SQ) was administered with local analgesia administered via exit site. Complete blood count and biochemistry were unremarkable incisional lidocaine before closure. During surgery, anesthesia was and culture and sensitivity of the exudate found a moderate growth maintained with isoflurane (2-4%) as needed and thermal support of Acinetobacter lwoffii. Fluoroscopy evaluation revealed restricted was provided via a recirculating warm water blanket. The entire inflation of lungs and a large, round, stationary cardiac silhouette. mass and overlaying skin was removed to achieve adequate margins Thoracic and abdominal ultrasound revealed lobulated pleural and the incision was closed with absorbable subcuticular sutures. effusion, no pericardial effusion, and free fluid in the abdomen. Due Recovery was uneventful, and meloxicam (0.125mg, PO) was to poor prognosis, euthanasia was elected. At necropsy over 700 administered. On cut surface, the mass was uniformly tan and milliliters of milky, white fluid was removed from the thoracic multilobular. The histomorphology was consistent with a cystic cavity, and severe bilateral atelectasis and abdominal effusion were papillary mammary carcinoma. At a 10-wk recheck, radiographs grossly appreciated. Fluid analysis of thoracic effusion reported showed no evidence of metastasis and FNA of cranial mammae/ predominately lymphocytes (94%), triglyceride and cholesterol lymph nodes showed no evidence of neoplasia. Surgical mastectomy counts of 873 mg/dL and 68 mg/dL, respectively, and no growth on should be considered in cases of mammary carcinoma of tree shrews. culture, confirming the diagnosis of lymphocytic-rich thoracic effusion, chylothroax. Vascular trauma from repeated attempts of PS12 Dracula the Laboratory Ferret advancing the intravenous catheter to the desired depth has been implicated in adhesion formation which eventually may lead to PO Mills*, JM Ritter failure of local lymphatic ducts. As chylothorax is a rare complication of indwelling catheters in rhesus macaques, this case emphasizes the National Center for Emerging and Zoonotic Infectious Disease, value of radiopaque catheter material and sufficient imaging Centers for Disease Control and Prevention, Atlanta, GA resources to confirm catheter placement locations and assist in follow up examinations. 693 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

PS14 The Tale of a Crooked Tail bacterial colonies were present. Differential diagnosis for the facial edema included irradiation damage to facial vessels, infection, or DR Webb-Wright*1, AE Brix3,1, TL Blankenship-Paris2 hypersensitivity reaction. In the following weeks, 2 additional mice developed facial edema and were euthanized. Further investigation 1Cellular and Molecular Pathology Branch, National Toxicology uncovered that the mice were irradiated in an empty microisolator Program, National Institute of Environmental Health Sciences cage. We hypothesize that due to the ability of the mice to rear, (NIEHS), Research Triangle Park, NC; 2Comparative Medicine multiple mice received increased doses of irradiation to the head, Branch, National Institute of Environmental Health Sciences causing vascular damage and secondary bacterial infection from (NIEHS), Research Triangle Park, NC; 3Experimental Pathology translocated gut bacteria. This phenomenon has been described in Laboratories (EPL) Inc., Research Triangle Park, NC one case study report, but facial edema is a noted sequela in many irradiation studies. For subsequent studies, the irradiation dose was A 30-d-old, male chimera mouse was examined for 2 discrete lowered, enrofloxacin replaced neomycin to provide broader swellings with dorsal deviations along the tail. The proximal swelling spectrum bacterial coverage, and a restraint device was used to was located at the tail base, with a narrowed, circumferential area of prevent rearing. No further clinical evidence of facial edema or apparent constriction caudally. The area of annular constriction was mortality were noted. This case emphasizes the differences between immediately followed by a second, more distal, fusiform shaped irradiation sources and their effects on dose dependent clinical signs. lesion with mild, dark blue to black subcutaneous discoloration, and a lesser degree of swelling and dorsal deviation. The differential PS16 Abdominal Mass and Thoracic Abnormality in a Rhesus diagnoses for these lesions included trauma, handling, nesting Macaque material entrapment, ring tail secondary to low humidity, improper tail vein injection(s), a subcutaneous abscess or cyst, a neoplastic MN Jackson*1, S Kirejczyk2, D Kempf1 process, or a combination of these things. The animal was bright, alert, in good body condition, and was a valuable founder, therefore, 1Department of Animal Resources, Yerkes National Primate Research the lesions were monitored. After 8 d, both lesions were larger in size, Center, Emory University, Atlanta, GA; 2Department of Pathology, the ventral aspect of the proximal lesion was ulcerated, and the Yerkes National Primate Research Center, Emory University, Atlanta, discoloration of the distal lesion was more pronounced. Following GA euthanasia, radiographs of the animal revealed a significant amount of soft tissue opacity in the areas of the swellings, caudal vertebrae A 19-y-old, indoor-housed female rhesus macaque (Macaca mulatta), malformations, and mild lysis of the dorsal aspect of the malformed with a previous history of 2 embryo transfers via laparotomy, was vertebrae. Additionally, a firm, raised, pale pink to tan nodule in the examined for an abdominal mass following a routine research MRI right cranial lung lobe was noted. All tissues were placed in buffered procedure. On physical examination, vitals were within normal limits formalin, and the tail was subsequently placed in decalcifying and an immobile firm mass was palpated within the caudal solution. Histopathology of both the tail and lung lesions revealed a abdomen. Bloodwork was unremarkable, 3+ blood was noted on mixture of tissues from different germ layers, including muscle, bone, urinalysis, and urinary retention was reported during the MRI scan. fat, squamous epithelium, keratin, and nervous tissue, leading to the Radiographs and ultrasound revealed a large, thick-walled, fluid- ultimate diagnosis of a malignant teratoma with metastasis to the filled mass in the caudal abdomen and increased soft tissue opacity lung. Teratomas are most commonly described in the reproductive in the left caudal lung fields. The mass was aspirated, and dark organs of animals and are usually benign. The literature describes brown, hemorrhagic fluid was collected. Differentials for the mass induced teratomas as a model for assessing pluripotency and included endometriosis, neoplasia, ovarian cyst, and possible tumorigenesis in research; however, this animal was experimentally hematoma. Differentials for the abnormality in the left thoracic space naïve. Although spontaneous neoplasia is less common in young included neoplasia and lung lobe atelectasis. Due to poor prognosis, animals, spontaneous teratomas are seen more often in younger the subject was sent to necropsy. Gross pathology revealed a 12cm- animals, and particularly in horses, strain 129, and genetically diameter, thick-walled cyst arising from the left ovary, multifocal modified mice. However, the extragonadal location of this teratoma, cysts along the gastrointestinal and diaphragmatic serosa, numerous and the presence of pulmonary metastasis makes this an unusual adhesions, and an anterior uterine/cervical leiomyoma compressing finding. the urinary bladder. Within the thoracic cavity, the left liver lobe and one-third of the greater omentum was herniated through a rent in the PS15 Facial Edema and Mortality in a Cohort of Irradiated B6.SJL- diaphragm, leading to atelectasis of the left caudal lung lobe. PtprcaPepcb/BoyCrCrl Mice Histology revealed the large cyst to be an endometrioma, with multiple endometriotic lesions throughout the abdominal cavity. KA Zabrecky*, PM Treuting, LE Neidig, SM Meeker Invasive endometriosis found within the membranous portion of the diaphragm likely led to the weakening of and fenestrations within Department of Comparative Medicine, University of Washington, the tissue, allowing for passage of abdominal viscera into the thoracic Seattle, WA cavity. Diaphragmatic hernia associated with endometrial lesions is a rare presentation for a well-known disease. Left undiagnosed, A cohort of B6.SJL-PtprcaPepcb/BoyCrCrl (B6.SJL) mice presented endometriosis in rhesus macaques can lead to various complications, with facial edema (8/40) and spontaneous mortality (5/40) 1 wk after thus routine monitoring and assessments are necessary to improve whole body x-ray irradiation. Mice were irradiated at a dose of 1200 the prognosis of high risk patients. rads and received 2 million bone marrow cells via retro-orbital injection containing either 1 million B6.129S2-Ifnar1tm1Agt/Mmjax or PS17 Anorexia and Weight Loss in a 19-y-old Olive Baboon (Papio B6.129P2(SJL)-Myd88tm1.1Defr/J cells with 1 million B6.SJL cells. anubis) Neomycin was administered prophylactically in the drinking water. This experiment had previously been performed without JA Herrod*, SE Beck, JM Izzi complication; however a cesium irradiator and C57BL/6J recipient strain were used. The affected mice were euthanized, and 2 were Molecular and Comparative Pathobiology, Johns Hopkins University, submitted for necropsy. On gross exam, severe facial edema and Baltimore, MD moderate segmental enteropathies were noted. Splenic cultures grew Enterococcus spp. Histopathology demonstrated multifocal A 19-y-old, 21 kg, individually housed male olive baboon (Papio moderate proliferative enteritis and typhlocolitis with villar blunting anubis) previously involved in imaging studies presented with a 2-wk and fusion and lymph node depletion, characteristic lesions post history of hyporexia. Routine physical exam performed 1 mo prior irradiation. In addition, diffuse edematous facial cellulitis with revealed moderate dental disease but was otherwise unremarkable. fibrinoid degeneration, necrosis of blood vessels, and abundant Examination at the time of presentation showed no change in

694 Abstracts of scientific papers 2019 AALAS National Meeting

severity of dental disease and a stable weight and body condition. PS19 Galactorrhea and Hepatomegaly in a New Zealand White Lab work revealed marked elevation of ALP (1,621 U/L) and GGT Rabbit (Oryctolagus cuniculus) (112 U/L), but all other blood parameters were within normal limits. Differential diagnoses included dental disease, or cholestasis caused RE Cooper*, C Lyons, SE Beck, JM Izzi by cholelithiasis, cholangitis, amyloidosis, neoplasia, or a gallbladder mucocele. Gastroprotectants, S-adenosylmethionine, and silybin were Molecular and Comparative Pathobiology, Johns Hopkins University, started along with pain medication and supportive care. Over the Baltimore, MD next 3 wk, there was no clinical improvement and additional diagnostics were pursued. An 8 percent decrease in body weight was A 3-y-old, multiparous, nongravid, individually housed female New noted. Abdominal ultrasound revealed multifocal dilated cystic Zealand White rabbit (Oryctolagus cuniculus) presented with multiple structures within the liver parenchyma and a dilated gallbladder swellings along the ventral abdomen. Physical exam revealed containing a moderate amount of sludge. No choleliths were seen uniformly enlarged, soft mammae of normal color and temperature. and the liver appeared of normal size and echogenicity. Repeat lab Milky discharge was easily expressible. Cytology showed prevalent work obtained at this time revealed worsening cholestasis (ALP: proteinaceous aggregates, as well as scant plasma cells, lymphocytes, 2,247 U/L; GGT: 191 U/L; Tbili 1.1mg/dL). Broad-spectrum monocytes, and neutrophils. Aerobic and anaerobic cultures were antibiotics, maropitant, and ursodeoxycholic acid were added to the negative for bacterial growth. Findings were consistent with normal treatment regimen. However, the baboon presented with jaundice milk. Abdominal palpation and radiographs were unremarkable. and anorexia 2 wk later and an exploratory laparotomy was Primary differential diagnoses at this time included mammary gland performed. During surgery, a thickened, firm, obstructed gallbladder hyperplasia in response to hyperprolactinemia (prolactinoma vs. was identified and euthanasia was elected due to poor prognosis. hyperestrogenemia [pseudopregnancy vs. uterine adenocarcinoma]), Histopathology revealed a densely cellular, poorly demarcated and and mammary gland adenocarcinoma. Over 5 wk, mammary infiltrative mass consistent with a gallbladder adenocarcinoma with enlargement persisted and body condition decreased. Abdominal secondary invasion into liver parenchyma. Hepatobiliary tumors radiographs and ultrasound exam at this time revealed comprise less than 6 percent of all neoplasms in baboons. Only 6 hepatomegaly with a rounded caudal liver margin. Complete blood cases of gallbladder adenocarcinoma have been reported, and this is count and serum chemistry showed a nonregenerative anemia, the first reported case in an olive baboon. increased hepatocellular enzymes, and hyperlipidemia; serum was grossly lipemic. Supportive care was initiated pending protocol- PS18 Episodic Fainting in a Castrated Male Domestic Short Hair planned euthanasia 1 wk later. Gross necropsy revealed an enlarged, Cat friable liver; histopathology confirmed hepatic lipidosis. The ultimate cause of hepatic lipidosis in this nonanorectic rabbit remains SM Young*2,3, E O’Connor1,2, C Ruggiero4, R Backus4 unknown, but prolonged lactation may have been a precipitating factor. Necropsy additionally revealed a normal pituitary gland, a left 1Office of Animal Research, University of Missouri, Columbia, uterine horn leiomyoma, and diffuse mammary gland hyperplasia. MO; 2Comparative Medicine Program, University of Missouri, Ovaries were normal with no corpora lutea. Previously reported Columbia, MO; 3Veterinary Pathobiology, University of Missouri, causes of leporine galactorrhea, including prolactinoma and Columbia, MO; 4Veterinary Medicine and Surgery, University of pseudopregnancy, were ruled out by these findings. Remaining Missouri, Columbia, MO possible causes of galactorrhea in this rabbit include ectopic hyperprolactinemia, pituitary-origin hyperprolactinemia (secondary A 13-y-old, 4.5 kg male castrated domestic shorthair cat on a vitamin to dopaminergic dysfunction, stress, or hypothyroidism), idiopathic D diet supplementation study presented with episodic syncope and hyperprolactinemia, or idiopathic galactorrhea. In humans, signs consistent with cardiogenic shock. Physical exam revealed pale leiomyomas have been identified as a source of ectopic and tacky mucous membranes, decreased peripheral pulse strength, hyperprolactinemia; uterine leiomyomas occur with some frequency mild dehydration estimated at 5-7%, rectal temperature of 95°F, a in rabbits, but galactorrhea as a comorbidity has not previously been heart rate of 160 BPM, and episodic fainting during examination. An reported in this species. intravenous catheter was placed and 50 mLs of lactated Ringer’s solution were administered. On auscultation, complete heart sound PS20 Head Tilt as a Result of an Unusual Etiology in a C57BL/6J deficits occurred during the periods of observed syncope. Three lead Mouse ECG revealed a left-bundle branch block with intermittent episodes of third degree atrioventricular (AV) block. Hematology, serum RJ Floyd*1, AO Michel1,2, A Piersigilli1,2, E Aronowitz3, H Voss3, RJ biochemistry, and ionized calcium levels did not reveal any Ricart Arbona1,2 significant abnormalities. Thoracic and abdominal radiographs and ultrasound revealed no cardiac abnormalities or signs of metastatic 1Tri-Institutional Training Program in Laboratory Animal Medicine disease. Echocardiogram showed no significant findings outside of and Science, New York City, NY; 2Center of Comparative Medicine complete ventricular standstill during periods of third-degree AV and Pathology, Memorial Sloan Kettering & Weill Cornel Medicine, block. Clinical signs of syncope occurred during periods of elongated New York City, NY; 3Citigroup Biomedical Imaging Center, Weill AV block. The cat was placed in an oxygen cage and observed Cornell Medicine, New York City, NY overnight with continuous cardiovascular monitoring. Clinical resolution of the cardiogenic shock and third-degree AV block A 2-mo-old female C57BL/6J mouse presented with a left-sided head occurred 48 h after the initial episode. The cat returned to his normal tilt. The mouse was otherwise normal. The animal, part of a cohort in housing, vitamin D2 supplementation was discontinued, and a study investigating the influence of inflammation on breast cancer continuous video monitoring was installed inside of the room. development and progression, had been provided a purified high fat Twenty-four-hour electrocardiographic (Holter) recordings (60%) diet (AIN-93) ad libitum since arrival from the vendor 2 wk performed 3 times at 4-w intervals revealed a progressive resolution earlier. All mice in the cohort were maintained in individually of the third-degree AV block and the cat remained clinically stable. ventilated cages with autoclaved aspen chip bedding, and acidified No cardiogenic or systemic cause of the AV block was determined, water. Differential diagnoses included idiopathic necrotizing arteritis, and clinical resolution occurred over time after termination of the bacterial otitis media/interna (Pasteurella pneumotropica, Pseudomonas vitamin 25(OH)D2 supplementation. Cardiac pacing was not aeruginosa, Streptococcus sp. Mycoplasma pulmonis, and Burkholderia implemented at this time due to the stabilized condition of the gladioli), encephalitis, abscess, neoplasia, congenital patient. The cat was removed from the study and was placed for malformation,and accidental or iatrogenic head trauma. Magnetic adoption. Currently the cat has remained stable with no apparent resonance imaging revealed a significant left sided displacement of cardiogenic deficits. the olfactory bulb and areas of hyperintense signaling consistent with edema. Following imaging, the animal was euthanized due to poor

695 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 prognosis. Histopathologic examination revealed a unilateral, full- multiple species. Recent recommendations suggest diets should be thickness bone defect at the base of the cribriform plate and a formulated with the more bioavailable form of vitamin K, malformation of the nasal conchae, resulting in the herniation of the phylloquinone (PK). Menaquinones (MKn) are produced by olfactory bulb into the nasal cavity. There was also a left midline-shift intestinal microbes and provide additional vitamin K through of the frontal cortex and moderate catarrhal sinusitis was present in coprophagy. Hypovitaminosis K was confirmed as no additional the left mandibular sinus. The MRI and histopathologic changes are morbidity was noted following treatment with 100 µg of PK suggestive of a congenital malformation of the nasal cavity and subcutaneously for 3 d and then every 3 d for 2 wk. The diet was also frontal aspect of the skull explaining the clinical presentation. reformulated with 1.2 mg/kg PK. Mice had significantly decreased Clinical, imaging, and pathological findings, prevalence in different MK4 in livers of antibiotic-treated mice prior to PK treatment (6+/- species and strains of mice, as well as the possible impact on research 0.9 pmol/g) compared to antibiotic-treated mice post PK (13 +/-1.2 will be discussed. pmol/g). Pre-PK fecal samples had significantly lower MK5-MK6 where MK7 and MK11 measured higher in antibiotic-treated mice PS21 White Growth on the Leg of a Xenopus laevis than in H. pylori-infected antibiotic-free mice. Fecal microbiome analysis in antibiotic treated mice pre-PK had significant decreases of ER Feldman*1, M Forzán2, RN Labitt1 bacteria in the phylum Bacteroidetes, many of which produce MKs, and an increase within the phylum Firmicutes. Post-PK treatment, the 1Center for Animal Resources and Education, Cornell University, microbiome of antibiotic treated mice rebounded where the profiles Ithaca, NY; 2Department of Population Medicine and Diagnostic matched H. pylori infected cohorts without antibiotics. Our data Sciences, Cornell University, Ithaca, NY demonstrates that marginal vitamin K in the diet when combined with antibiotic-induced intestinal dysbiosis, induced clinical vitamin A 9-year-old female Xenopus laevis was reported for a white growth K deficiency. on its left leg. On physical exam, the frog was quiet and had a white, cotton-like mass on the left tarsus. Both eyes had white corneal PS23 Administration of Oral Diclofenac Results in Signs of NSAID opacities that were easily irritated and bled freely. Cytology of the Toxicity in Rats Harboring the Human ATG16L1 Crohn’s Disease growth revealed abundant fungal hyphae, which were identified Susceptibility Variant as Saprolegnia parasitica via fungal culture. Due to the advanced age of the frog and poor condition, euthanasia was elected. On gross KL Chesney*1,2, EC Bryda1,2 examination, there was an approximately 6 mm x 2 mm white skin defect with hyperemia where the growth was previously present on 1Comparative Medicine Program, University of Missouri, Columbia, the left tarsus. The spleen was severely enlarged and mottled dark MO; 2Veterinary Pathobiology, University of Missouri, Columbia, red with a white, fibrous capsule surrounding the serosal surface. MO The liver and heart were also enlarged and discolored. The spleen and eye were submitted for fungal culture, which yielded no growth. Crohn’s disease (CD) is 1 of 2 chronic inflammatory bowel diseases Histology revealed populations of round neoplastic cells, forming (IBD) that affect the lining of the gastrointestinal (GI) system. Several sheets and perivascular infiltrates in the spleen, liver, kidney, environmental factors, either through acute insult or chronic build- digestive mucosa, blood vessels, eye, and skin. up over time, contribute in large part to the multifactorial etiology of Immunohistochemistry showed antibodies against CD3 cells this disease. For example, using a knock-in rat model harboring an colocalizing with neoplastic cells in the spleen and liver, identifying ATG16L1 gene variant responsible for increased CD predisposition in them as T cells. Based on gross and histological findings, the final humans, our laboratory previously found that longterm, low-dose diagnosis was a multi-organ, T cell lymphoma. Saprolegnia commonly exposure to the nonspecific, nonsteroidal antiinflammatory (NSAID) occurs in amphibians secondary to stress or disease. In this case, the diclofenac (1.25 mg/kg, 50% of the no-observable-effect level frog was likely immunocompromised due to lymphoma, resulting in (NOEL)) results in shifts in the gut microbiota composition as well as colonization by Saprolegnia. The presence of Saprolegnia warrants CD-like histologic signs in ileal and colonic tissues of rats investigation into both environmental and pathological causes of heterozygous for the Atg16l1 CD susceptibility variant (HET). To immunocompromise in Xenopus laevis. assess whether acute exposure to the same NSAID could also trigger disease onset, HET rats (4 male, 4 female) from the Atg16l1 strain and PS22 Hypovitaminosis K in Mice: Do Fecal Menaquinones their wild type (WT) littermates (4 male, 4 female) were orally Overcome Dietary Insufficiencies? gavaged with the NSAID diclofenac at 10 mg/kg once per day for 7 d to simulate a shortterm course of high-dose NSAID administration LA Quinn*1, A Sheh1, HR Holcombe1, J Ellis2, D Smith2, S Booth2, X in humans. We found that only HET rats displayed overt signs of Fu2, S Muthupalani1, Z Ge1, D Puglisi1, TA Gonda3, T Wang4, JG Fox1 disease as well as gross and histologic signs of NSAID toxicity and CD-like lesions at necropsy, with females showing more severe signs 1Department of Comparative Medicine, Massachusetts Institute of of disease than males. Gross signs of disease included pale livers Technology, Cambridge , MA; 2Human Nutrition Research Center on with multifocal necrosis, pale kidneys, and jaundiced, fibrotic Aging, Tufts University, Boston, MA; 3Division of Digestive and Liver omentum. Histologic signs included multifocal liver necrosis and Diseases Department of Medicine, Columbia University, New York, multifocal areas of lamina propria inflammation, mucosal erosion, NY; 4Columbia University, New York, NY and transmural ulceration of the ileum and colon. These results indicate that rats harboring the Atg16l1 variant causing increased We previously published that antibiotic treatment or increased susceptibility to CD in humans are more sensitive to diclofenac dietary folate significantly decreases gastric pathology in Helicobacter treatment than their WT littermates. Further studies are needed to pylori-infected mice. To determine if combining the treatments would elucidate the mechanism underlying why this genetic variant causes act synergistically, cohorts of male INS-GAS mice infected with H. increased sensitivity to NSAID administration resulting in CD-like pylori (SS1) were administered antibiotics and/or high-dietary folate histologic lesions. (8 mg/kg vs 2 mg/kg diet) at 18 wk postinfection. Antibiotics, but not high folate, decreased gastric disease severity. However, PS24 Exploring a Light-based Modality for in Vivo Antimicrobial unexpected morbidity and mortality associated with acute blood loss Treatment of the Brain was observed in 41% of infected mice receiving antibiotics, and in 8% of infected mice not receiving antibiotics. Our presumptive diagnosis was hypovitaminosis K due to dietary insufficiency combined with C Thurman*1,2, A Muthuswamy3,2, M Klinger1,2, G Roble4,2 antibiotic-induced dysbiosis. Mice were fed a chemically defined amino acid diet with menadione (MD) as the form of vitamin K. MD 1Office of Veterinary Resources, New York University, New York, is a synthetic, water-soluble form of vitamin K used in diets for NY; 2Post-Doctoral Program in Laboratory Animal Medicine, NYU-

696 Abstracts of scientific papers 2019 AALAS National Meeting

Regeneron, New York, NY; 3Regeneron Pharmaceuticals, Tarrytown, by PMSG or AIS superovulation methods. Further embryo transfer NY; 4Comparative Medicine, Fred Hutchinson Cancer Center, Seattle, experiments will be performed to determine the numbers of viable WA pups that were derived from vitrified-warmed PMSG or AIS superovulation methods. Antimicrobial resistance is a growing problem in human medicine that extends to biomedical research. Light-based therapies present an PS26 Improvement of Retinal Function in Diabetic Rats After alternative to traditional pharmaceuticals and are less vulnerable to Subconjunctival Injection of Insulin-loaded Nanogels acquired bacterial resistance compared to chemical-based therapies. The brain poses its own set of difficulties with respect to CV Fili*1, FF Sahle2, DJ Hamilton1, TL Lowe2 antimicrobial therapy due to immunological privilege and relative tissue sensitivity to topical antibiotics. This study focused on 405nm 1Comparative Medicine, University of Tennessee Health Science “true violet” light —which has been shown to kill multiple clinically Center, Memphis, TN; 2Pharmaceutical Sciences, University of relevant bacterial species in vitro while leaving mammalian cells Tennessee Health Science Center, Memphis, TN unscathed—and its effect on the murine brain. This study was carried out in 3 parts. First, we built a 405nm LED (light-emitting diode) Diabetic retinopathy (DR) is the most common neurovascular array and validated its power using an optical sensor. Then, we complication of diabetes, affecting approximately 140 million tested in vitro efficacy against a clinical bacterial isolate (Enterobacter diabetics worldwide. Current treatment options, including laser cloacae) from a nonhuman primate open craniotomy site and vitrectomy and anti-VEGF agents, are only used for advanced DR demonstrated bacterial attenuation at our selected doses to validate and have a high non-response rate, many side effects, and do not antimicrobial efficacy. Finally, we treated 84 randomized male and address the underlying neuroretinal pathology of DR. To target the female C57Bl/6J and C57Bl/6CRL mice at the time of craniotomy neuronal degeneration in DR, a sustained release, insulin-loaded, with various doses of light (0, 36 J/cm2, 45 J/cm2, 54 J/cm2) to assess biodegradable nanogel system was developed as a potential long- safety of the device in vivo. Mice were observed daily until term therapeutic to rescue retinal neurons from apoptosis and reduce euthanasia at 1, 3 or 21 days postsurgery. The selected doses caused DR onset and progression. The purpose of this project was to neither behavioral changes postoperatively, as measured by cageside determine if a single subconjunctival injection of these nanogels observation for pain behaviors or neurologic abnormalities, nor would improve retinal function in diabetic Sprague Dawley rats as observable pathology to the brain, as determined postmortem by measured by electroretinogram (ERG). Insulin was loaded in the histological evaluation and immunofluorescence staining nanogels during nanogel synthesis with over 98% loading efficiency. for Cas3 and GFAP, markers of apoptosis and necrosis. True violet The in vitro release of insulin from the nanogels was studied by light devices may present an inexpensive refinement to current dialysis method, and the released insulin was quantified by ultra practices for maintaining open craniotomy sites or reducing bacterial performance liquid chromatography. The nanogels sustained the loads in contaminated surgical sites. Further studies will be needed release of insulin in vitro for at least 60 days. Male Sprague Dawley to assess in vivo bacterial attenuation in the brain either in active rats (n=6) were induced to become diabetic with a single clinical cases or by controlled inoculation. intraperitoneal injection of 50 mg/kg of streptozotocin in citrate buffer. Age-matched control rats (n=6) received citrate buffer alone. PS25 In Vitro and in Vivo Developmental Comparison of Vitrified- One month after diabetes induction, diabetic rats received a single Warmed C57BL/6 Zygotes Derived from Anti-Inhibin Serum or subconjunctival injection of 20 µL of insulin-loaded nanogels in the Pregnant Mare Serum Gonadotropin Superovulation left eye and 20 µL of phosphate buffered saline in the right eye. Scotopic ERG measurements were performed on both diabetic and S Gerb*, Y Agca, C Agca non-diabetic control rats after two hour dark adaptation one day before, one week after, and two weeks after treatment. The ERG data Veterinary Pathology, University of Missouri, Columbia, MO showed that a-wave and oscillatory potential amplitudes were significantly increased two weeks after treatment with the nanogels Cryopreservation of germplasm such as sperm and embryos is an compared with one day before treatment (p<0.05) and were important and cost effective method to preserve scientifically increased to amplitudes similar to those in the naive control rats. important rodent models of human diseases and safe guard them These results suggest that the insulin-loaded nanogels improved against genetic drift, contamination, infectious disease, and natural photoreceptor and amacrine cell function in diabetic rats and can be disasters. Currently, the gold standard consists of superovulation potentially developed as a long-acting therapy for the treatment and with Pregnant Mare’s Serum Gonadotropin (PMSG). Recent prevention of diabetic retinopathy. investigation has shown that superovulation of mice by anti-inhibin serum (AIS) yields significantly higher oocytes compared to PMSG PS27 Effects of Carprofen and Buprenorphine on Tumor Growth in method which reduces the number of donor females. However, there Mouse Models of Prostate Cancer Bone Metastasis is less information available about the cryosurvival of AIS derived embryos. In this study, 10 C57BL/6 female mice were superovulated JJ Xu*1, T Robinson2, S Thurston1, HE Thomas1, S Daignault3, JF with either AIS or PMSG. Clutches of metaphase II oocytes were Escara-Wilke2, ET Keller1,2, JM Keller1,2 collected from the oviducts following hCG administration, and inseminated via in vitro fertilization. A subset of fresh embryos were 1Unit for Laboratory Animal Medicine, University of Michigan, Ann immediately cultured to assess their developmental potential to Arbor, MI; 2Department of Urology, University of Michigan, Ann blastocyst stage. Remaining embryos derived from either AIS or Arbor, MI; 3School of Public Health, University of Michigan, Ann PMSG were cryopreserved via vitrification and later warmed to Arbor, MI determine both in vitro and in vivo developmental competence. The percentage of fresh embryos that developed to the blastocyst stage Murine models of tumor development often require invasive from PMSG (85.75±2.09) and AIS (84.30±1.87) mice were not different procedures for implantation, potentially causing pain or distress. (p>0.05). The percentage of the vitrified zygotes that were intact after However, analgesics are often withheld because they may affect warming for PMSG (93.3±2.06) and AIS (89.3±1.78) mice were not tumor development. Previous studies examining the effects of different (P > 0.05). The percentage of vitrified-warmed embryos that analgesics on the development and metastasis of various tumor lines developed to the blastocyst stage from PMSG (53.33±1.97) and AIS show that the effect of analgesics on the tumors is dependent on (56.66±1.71) mice were also not different (P > 0.05). Vitrified-warmed tumor line and analgesic used. A blanket statement that analgesics zygotes obtained from either AIS or PMSG were cultured to 2-cell affect the general growth of tumors is not adequate scientific and surgically transferred into CD-1 surrogate females to compare justification for withholding analgesics, and pilot studies or pregnancy rates. Equal pregnancy rates were obtained for PMSG references are recommended for each specific scenario. In this study, (3/3) and AIS (3/3) groups, indicating that embryo quality is similar we evaluated the effects of two commonly used analgesics on tumor

697 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 growth in two models of prostate cancer (PCa) bone metastasis. We PS29 Depletion of CD4 T-cells Increased Epstein-Barr Virus hypothesized that a one-time injection of analgesics at the time of Infections in a New Zealand White Rabbit (Oryctolagus cuniculus) intratibial injection of PCa cells would have no effect on tumor Model growth. C57Bl/6 or SCID mice were injected subcutaneously with an analgesic (carprofen 5 mg/kg or buprenorphine 0.1 mg/kg) or AJ Osborne*1, K Balogh2, S Brendle2, J Hu2, C Sample3, ND vehicle (saline 0.1 ml) at the time of intratibial injection with PCa cell Christensen2 lines (RM1 or PC3, n=10 per group). Tumor progression (bioluminescent imaging, radiographs), behavioral assays (Von Frey), 1Comparative Medicine, Penn State Hershey, Harrisburg, and clinical signs (body weight) were monitored for 2-4 weeks. PA; 2Pathology, Penn State Hershey, Hershey, PA; 3Microbiology and Neither carprofen or buprenorphine administration affected tumor Immunology, Penn State Hershey, Hershey, PA growth, behavioral or clinical parameters compared to the saline control for either cell line. Overall, this study adds to the growing Epstein-Barr virus (EBV) is a gamma-herpesvirus that infects over body of literature demonstrating that animal welfare can be 90% of the adult human population, is the main etiologic agent compatible with scientific objectives, and the decision to withhold of infectious mononucleosis, and is associated with cancers such analgesics must be evaluated on a case-by-case basis. as Hodgkin’s or Burkitt lymphomas. EBV has been studied in lab animals using EBV-like viruses, but an adequate animal model has PS28 A Comparison of Ketamine and Etomidate-based not yet been established. The goal of this project was to compare Intraperitoneal Anesthetics in Multiple Mouse Strains two immunosuppressive regimens, Cyclosporine A (CsA) and CD4 T-cell antibody depletion to establish a rabbit model of EBV. Six adult C Gergye*, Y Zhao, V Lee, R Moore NZW rabbits were inoculated with EBV via the marginal ear vein. Two rabbits (female =1, male =1) were treated with 15 mg/kg CsA Emory University, Atlanta, GA subcutaneously at the time of EBV inoculation. CsA treatments were continued at 15 mg/kg daily for four days, and then at 20 mg/kg Intraperitoneal (IP) injection is a common route of anesthetic twice weekly for two weeks. Four rabbits (female = 2, male = 2) were administration in mice, however few safe and effective protocols treated with 2 mg anti-CD4 T-cell antibody IV concurrent to EBV have been developed. Ketamine-xylazine IP anesthesia is one of the inoculation and then once a week for two weeks. Peripheral blood most widely used IP protocols, but has limitations in efficacy and lymphocytes (PBL) and body temperatures were monitored until suitability for some studies. Etomidate is an alternative to ketamine study endpoint. Both rabbits in the CsA group and one rabbit in the that is used in human and companion animal practice that has not anti-CD4 group showed signs of illness such as hyporexia, decreased been widely explored in mice. We evaluated etomidate-xylazine (EX) fecal output, and lethargy; these rabbits reached humane endpoints IP anesthesia as an alternative to ketamine-xylazine (KX) anesthesia. despite veterinary supportive care. Partial depletion (average 12% We hypothesized that EX anesthesia would be as safe and effective decrease from baseline) of CD4 T-cells was demonstrated by flow as KX anesthesia, with anticipated sex- and strain-dependent cytometry. qPCR analysis of spleen cells proved more diagnostic differences. Male and female Crl:CD1(ICR) (n=42), C57BL/6NCrl than PBLs, and the anti-CD4 rabbits were strongly positive for (n=34), BALB/cJ (n=37) and NU/J (n=34) mice were given a single EBV within splenic cells whereas the CsA-treated rabbits were IP dose of ketamine 100 mg/kg and xylazine 10 mg/kg or etomidate weakly positive. Future aims for this model include optimizing the 20 mg/kg and xylazine 10 mg/kg. Multiple physiologic parameters, CD4 T-cell antibody depletion regimen with the goal of 100% CD4 anesthetic time points, and mortality rate were evaluated. Surgical depletion to standardize immunosuppression. The rabbit EBV model depth was confirmed by a negative pedal withdraw response to requires further characterization, but is a promising model to test both manual finger pinch and regulated forceps pinch. Sedation anti-viral medications and prophylactic vaccines for EBV in patient time, defined by duration of loss of righting reflex, was similar populations. between KX and EX anesthesia with CD1 mice exhibiting shorter sedation times regardless of protocol. Several mice experienced an PS30 Placement and Use of an Indwelling Epidural Catheter with adverse hyperexcitement response during induction, with BALB/ Concurrent Behavioral Assessment in Yucatan Minipigs (Sus cJ and NU/J mice administered EX significantly more likely to scrofa) experience hyperexcitement. Surgical anesthesia was achieved in 43.84% of EX animals compared with 4.17% of KX animals. 93.75% KT LaVallee*1, MD Unger2, JA Scholz1, TP Maus3, AS Beutler2 of C57BL/6NCrl mice administered EX lost pedal withdraw reflex and were significantly more likely to achieve surgical anesthesia 1Comparative Medicine, Mayo Clinic, Rochester, MN; 2Oncology, when compared to other strains. Venous pH was evaluated 40 min Mayo Clinic, Rochester, MN; 3Radiology, Mayo Clinic, Rochester, MN post-IP injection for all mice. Male NU/J were significantly more likely to exhibit moderate-severe acidosis as compared to males of Image–guided percutaneous spine procedures offer an attractive other strains. Mortality rates were low for both protocols, with no approach to treating spinal targets because minimally invasive mortalities noted during KX administration and 1 mortality after techniques are well-tolerated and do not necessitate general EX administration. Overall, these results indicate that EX is a more anesthesia. A useful preclinical model of this therapeutic modality effective surgical anesthetic than KX. However, due to multiple would provide the means to test tolerability of novel spinal agents factors such as adverse reactions, strain- and sex- associated delivered to conscious animals in a species that exhibits homology differences, EX IP anesthesia may only be acceptable as a safe and with the structure and scale of adult human spinal anatomy. We set effective alternative to KX anesthesia in C57BL/6NCrl mice. The out to establish a model of epidural infusion in 3 conscious Yucatan variability among mouse strains and between sexes requires further swine to assess the feasibility of catheter maintenance and repeat use investigation to optimize IP EX dosage. in an open field arena over 2 wk. CT-fluoroscopic guidance was used for percutaneous epidural catheter placement in each anesthetized animal with concurrent carprofen analgesia. Behavior was assessed prior to catheter placement and during vehicle infusions (beginning 3 d postcatheter placement) using a modified visual analog scale (VAS) by 2 blinded veterinarians. VAS parameters included restlessness, vocalization, posture, ambulation, injurious behavior, and overall clinical pain. Bolus infusion of epidural lidocaine was available as rescue analgesia. All catheters remained in place throughout the study, demonstrated on CT imaging (at 1- and 2- wk postcatheter placement) by exclusive epidural contrast flow and stability of spinal segments infused. Clinical exams remained within normal

698 Abstracts of scientific papers 2019 AALAS National Meeting

limits throughout the study and lidocaine rescue was not required prominent and tortuous surface vessels. Histologically, lesions at any timepoint. Necropsy and gross examination at 4 wk did not were characterized by dilated spaces surrounded by a thin fibrous reveal evidence of infection, inflammation, or fibrosis of neuraxial capsule often accompanied by early fibroplasia and/or edema tissues. Interrater reliability of the modified VAS was strong. No (7/10). Cystic spaces were lined by cuboidal (10/10) to columnar significant difference in animal behavior was observed between (7/10) epithelium. In all cases there was lymphocytic infiltration vehicle infusion and control timepoints. These results suggest that of the fibrous capsule, and most cases also had neutrophilic (8/10) indwelling epidural catheterization in conscious swine is feasible and and histiocytic (9/10) infiltrates. There was occasional hemorrhage safe for a period of at least 2 wk. As a model of adult human spinal (5/10) and hemosiderin-laden macrophages. Lesions were also intervention, swine offer a robust platform to test tolerability of novel characterized by varying degrees of pancreatic acinar atrophy (6/10) agents intended for use in the awake patient. and fibrosis (5/10). Animals were predominantly, but not exclusively, of NOD and NOD-derived backgrounds. Four of 10 animals were PS31 Comparison of Injectable Anesthetic Management Protocols NOD.Cg-PrkdcIL2rg/SzJ [aka NOD scid gamma, for Swine (Sus scrofa domestica) Utilized in a Paramedic or NSG] mice. Four of 10 animals were from the following strains Emergency Tracheostomy Teaching Laboratory (one of each): NOD/ShiLtJ [NOD], NOD.CB17-Pkrdc/J J Brune*, N Reyes [NOD scid], NOD.Cg-Prkdc Il2rg Tg(CMV- IL3,CSF2,KITLG)1Eav/MloySzJ [aka NSG-SGM3, or NSGS], and Comparative Medicine, University of Washington, Seattle, WA NOD.B-(D3Mit93-D3Mit124)(D4Mit114-D4Mit142)/1112MrkJ [NOD. c3.c4]. Two of 10 animals were not NOD-derived strains: BTBR.V(B6) We report comparison of two injectable anesthetic protocols utilized Lep/WiscJ [BTBR obese] and CBySmn.CB17-Prkdc/J in adult male and female 20-30kg domestic swine for a terminal [BALB scid]. None of these cases had gross or histologic evidence of teaching laboratory in which paramedics conduct an emergency luminal obstruction, and the ultimate driver of these cystic dilations tracheostomy. The laboratory requires the use of injectable anesthetic could not be definitively determined. Although regarded as rare, to prevent exposure of participants to inhaled anesthetics and pancreatic cystic lesions should be considered as a differential to maintain a stable plane of anesthesia as the trachea is incised diagnosis for laboratory mice with abdominal distention, and the and intermittently obstructed over the course of the lab. An ideal lesion may be overrepresented in NOD and NOD-derived strains. protocol should provide a stable anesthetic plane, cause minimal aberrations in cardiac function, be uncomplicated to administer, PS33 Development of a Fistula into the Cecum of a Deer Mouse cost effective, and utilize readily available drugs. In both protocols, (Peromyscus maniculatus) Post Castration Surgery swine were premedicated with intramuscular midazolam 0.5 mg/ kg, butorphanol 0.3 mg/kg and xylazine 4 mg/kg followed by V Smid* isoflurane administration via nose mask, intubation and intravenous catheter placement. All swine received intravenous buprenorphine Canadian Food Inspection Agency, Winnipeg, Canada 0.01 mg/kg prior to surgery. At the start of the lab, isoflurane was discontinued. One group of swine (n=4) received a bolus of Male deer mice were castrated to remove the influence of alfaxalone 0.47 mg/kg followed by 6 mg/kg/hr continuous rate testosterone on the variable of study with testosterone being replaced infusion. A second group of swine (n=4) received 1 ml/kg bolus of via administration through an osmotic pump to control dosage 5% guaifenesin 0.1% ketamine and 0.1% xylazine (GKX) followed and timing.Surgical castration was performed under isoflurane by a 2.2 mL/kg/hr continuous rate infusion. Vitals including heart anesthesia using aseptic technique. A 2-incision method was rate and non-invasive blood pressure were monitored every 10 used, incising over each testicle in the scrotum, exteriorizing the minutes. The relative stability of cardiovascular function of the testicle, clamping and ligating the vessels and spermatic cord with two protocols was determined by comparison of the average slope absorbable suture followed by closure of the skin with tissue glue. for linear regression functions for the mean arterial pressure and Mice were administered meloxicam at 2 mg/kg once every 24 h and heart rate. No significant difference was observed between the two enrofloxacin at 10 mg/kg subcutaneously just prior to surgery and protocols in either parameter. However, the GKX group experienced for at least 2 more d afterward every 24 h. Treatment with analgesia relative higher mean arterial pressures and heart rates trended lower and/or antibiotic was extended if required based on monitoring compared to the alfaxalone group. In conclusion, both protocols observations including but not exclusive to active inflammation, were comparable in providing a stable surgical anesthetic plane, cost discomfort, or reddness at the incision site. One mouse eventually effectiveness, ease of administration and cardiovascular stability. presented with a complication postsurgery that would not become The differences in mean arterial pressure and heart rate between evident until approximately 2 mo after the initial procedure was protocols can be attributed to the action of xylazine, an alpha-2 performed. The 2-mo-old mouse had no complications noted agonist, in the GKX protocol. Ultimately, both protocols present during surgery. On d 5 post surgery, a small amount of reddish, viable options for use in swine and can be used preferentially based clear discharge was observed from the right-side scrotal incision. It on drug availability and targeted length of anesthesia. was treated with topical chlorhexidine ointment and enrofloxacin injectable. Enrofloxacin was continued until d 11 post surgery. PS32 Pancreatic Cystic Lesions in Laboratory Mice from The mouse continued eating or drinking; body weight and body Predominantly NOD and NOD-derived Backgrounds condition as well as activity level remained normal. On d 56 post surgery, a brown discharge was observed from the right scrotal JG Vilches-Moure*1, DM Imai2, R Doty3 incision area. Under isoflurane anesthesia, the area was cleaned with chlorhexidine and flushed with saline. This procedure was repeated 1Comparative Medicine, Stanford University, Stanford, for approximately 3 more d until it was observed that the discharge CA; 2VM:PMI, Comparative Pathology Laboratory, Davis, resembled ingesta. The mouse was euthanized and a postmortem CA; 3Pathology Services, The Jackson Laboratory, Bar Harbor, ME exam revealed a fistula formation between a previous abscess and the cecum through the body wall of the mouse. Despite this fistula Reports in the literature of pancreatic cysts in mice are generally formation, the mouse had shown no other signs of illness other associated with polycystic disease (such as polycystic kidney than the noted brown discharge in the previous few days. This case disease), or as a single feature of the overall lesions observed highlights possible unusual sequelae to a relatively straightforward in pancreatic cancer models. However, pancreatic cystic lesions surgical procedure as well as the time that can elapse before that in mice outside those scenarios are exceedingly rare. Here, we sequelae becomes evident. report a series of 10 purpose-bred laboratory mice with pancreatic cystic lesions. Generally, animals were submitted to diagnostic necropsy for abdominal distension and/or a palpable abdominal mass. Grossly, cystic lesions were thin walled and often had

699 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

PS34 Unanticipated Disease Severity in the IL10-/- Mouse Model of standard chow diet was reinstituted. The remaining mice were Inflammatory Bowel Disease administered subcutaneous fluids and intraperitoneal dextrose for 2 d, and offered palatable nutritional supplements for 1 wk. Following SM Young*1,2, CL Franklin1,2 diet change and supportive care, there were no additional morbidities or mortalities in the colony. Surveillance testing 4 wk 1Veterinary Pathobiology, University of Missouri, Columbia, later by PCR of pooled fecal samples was positive MO; 2Comparative Medicine Program, University of Missouri, for Helicobacter spp. and Entamoeba spp. Surviving mice have been Columbia, MO clinically normal for 3 mo and breeding has commenced. In conclusion, sudden changes in the diet fed to wild- The IL10-/- mouse is used as a model to study inflammatory bowel caught Peromyscus may lead to anorexia and acute onset of hepatic disease. An optimal phenotype requires postweaning oral gavage lipidosis. These cases highlight the importance of gradual dietary with Helicobacter hepaticus, which serves as a provocateur of an changes combined with close observation of wild-caught Peromyscus inflammatory response to intestinal microbiota. Mice not inoculated leucopus housed in laboratory settings. with H. hepaticus develop sporadic and less severe disease. A researcher using this model observed an increase in the severity of PS36 Clinical Hypovitaminosis K in Mice Secondary to Antibiotic- disease (rectal prolapse, weight loss, histologic lesion scores) of IL10- induced Dysbiosis and Marginal Vitamin K Levels in a Chemically /- saline-gavaged control mice that did not receive H. hepaticus. The Defined Diet concern for increased disease in control experimental mice was expressed after the colony was relocated to a modular unit during LA Quinn*1, HR Holcombe1, A Sheh1, J Ellis2, D Smith2, S Booth2, X vivarium renovation. In order to investigate the cause of the Fu2, S Muthupalani1, Z Ge1, D Puglisi1, T Wang3, TA Gonda3, JG Fox1 increased disease occurrence in the control mice, metagenomic evaluation of feces, cecal, and mid-jejunal contents was performed to 1Department of Comparative Medicine, Massachusetts Institute of look for a potential dysbiosis, and intestinal tissues were evaluated Technology, Cambridge, MA; 2Human Nutrition Research Center on by histology. Metagenomics data revealed no dysbiosis, Aging, Tufts University, Boston, MA; 3Columbia University, New histopathology revealed inflammation, and epithelial hyperplasia York, NY present in the cecum and colon consistent with mild to moderate disease. Metagenomic analysis unexpectedly identified bacteria in A study was conducted to assess potential therapeutic synergy in the genus Helicobacter present in the mid-jejunal contents of all treating Helicobacter pylori-infected male INS-GAS mice with control mice analyzed. A PCR to screen for H. hepaticus was then increased dietary folate supplementation and antibiotic combination performed in the remaining breeding animals in the colony, and eradication therapy. Mice were infected with H. pylori (SS1) and fed a positive results for H. hepaticus were obtained. The increase in disease chemically defined amino acid diet containing 2 mg folate/kg. severity of the control mice was contributed to contamination with H. Eighteen weeks postinfection (WPI), specific cohorts were switched hepaticus in the colony breeders and stock animals. The timeline of to an 8 mg/kg folate diet and/or received helicobacter eradication contamination was determined by PCR evaluation of historical therapy consisting of omeprazole (400 μmol/kg/day), metronidazole paraffin embedded tissues. The colony was reestablished, and (14.2 mg/kg/day), and clarithromycin (7.15 mg/kg/day) in a 0.2-mL routine screening of mice was implemented. This occurrence stresses volume orally twice a day for 7 d. Mice that received the eradication the importance of pre-inoculation screening to ensure negative status therapy became lethargic and appeared scruffy, hunched, and pale of experimental pathogens. within 4 d after completion of antibiotic treatment. Starting at 19 WPI, 41% of the mice that were given antibiotics were found dead or PS35 Hepatic Lipidosis following Abrupt Diet Change in Wild- moribund. Animals that were moribund were euthanized with

caught White-footed Mice (Peromyscus leucopus) CO2 and they, along with the mice found dead, were necropsied. On gross examination mice had pale tissues, a pale and friable liver, and CS McCoy*1, S Carrasco1, RM Kramer1, JL Haupt1, J Buchthal2, S a stomach filled with unclotted blood. Complete blood cell counts Muthupalani1, JG Fox1 confirmed severe anemia due to acute whole blood loss. At 24 WPI, 2 mice on 8 mg/kg folate diet that did not receive antibiotics were also 1Division of Comparative Medicine, Massachusetts Institute of found dead with the same findings on necropsy. Our tentative Technology, Cambridge, MA; 2Media Lab, Massachusetts Institute of diagnosis was hypovitaminosis K. The source of vitamin K added to Technology, Cambridge, MA both the 2 and 8 mg/kg folate diet was 0.5 mg menadione sodium bisulfite (MD) per kg of feed. Previous studies question the Twenty adult male and female white-footed mice (Peromyscus bioavailability and optimal feed concentration of menadione in leucopus) caught near Cape Cod were socially housed by sex in a animal feed and recommends the use of phylloquinone (PK). Fecal barrier room. Quarantine testing by PCR of pooled fecal samples was and liver samples were collected for vitamin K level analysis. In positive for Cryptosporidium spp.,Helicobacter spp., fur mites, and antibiotic treated mice, liver MK4 was significantly decreased pre-PK pinworms. After 14 wk, the diet was changed from a standard rodent treatment (6+/-0.9 pmol/g) compared to post-PK (13 +/-1.2 chow to a medicated rodent chow containing 150 ppm fenbendazole, pmol/g). All surviving mice were treated with 100 µg of PK 25 ppm pyrvinium pamoate, and 1000 ppm doxycycline. Five days subcutaneously for 3 d and then every 3 days for 2 w. Within 24 h of later, an adult male was found dead with no premonitory signs. PK treatment, the mice had improved body condition scores and Despite increased monitoring, 2 mice were found dead, and 3 activity. The diets were reformulated with 1.2 mg PK/kg. No moribund mice were euthanized 3 d later. One moribund mouse additional morbidity or mortality occurred for the remainder of the exhibited severe neurologic signs, with lateral recumbence, study when animals were euthanized with CO2 at the 28 WPI opisthotonus, and paddling. Gross pathologic findings of all mice predetermined end point. included thin body condition, empty gastrointestinal tracts, and atrophy of subcutaneous and intracavitary fat. Livers were PS37 Urinary Tract Infection Associated with Bladder moderately enlarged, diffusely pale-tan, and friable. On Augmentation Surgery in Baboons (Papio anubis) histopathological examination, livers had severe macro- and micro- vesicular hepatic lipidosis. The mouse exhibiting neurologic signs S Oh*, L Halliday had chronic proliferative gastritis with intralesional eggs and adult nematodes consistent with Capillaria spp. The white matter at the Biologic resources Laboratory, University of Illinois, Chicago, IL levels of the hippocampus and cerebellum in this mouse exhibited multifocal areas of spongiosis with mild laminar neuronal loss, and Bladder augmentation is a treatment option to increase bladder astrocytosis with Alzheimer type II cells. These changes are volume in situations that cause a decreased bladder capacity. suggestive of hepatic encephalopathy. After the second mortality, the Ileocystoplasty is the standard technique for bladder augmentation.

700 Abstracts of scientific papers 2019 AALAS National Meeting

However, it has been associated with urinary tract infections (UTI), urolithiasis, intestinal obstruction, and long-term risk of bladder PS39 Clinical Cytomegalovirus in a Naïve Subadult Rhesus cancer. Various biomaterials have been proposed as an alternative to Macaque (Macaca mulatta) increase bladder size while minimizing complications. The baboon (Papio anubis) has been developed as a bladder augmentation model, CK Wallace*1, L Makaron1, R Wolf3, G Adams2, S Heidi2 but there is a paucity of information with regards to complications. This case series reports the incidence of UTI following bladder 1University Laboratory Animal Resources, University of augmentation in baboons. A retrospective review of 24 patients (12 Pennsylvania, Philadelphia, PA; 2Neurosciences, University of males, 12 females) was performed. There were 4 groups; cystectomy Pennsylvania, Philadelphia, PA; 3Radiology, Neuroradiology section, (n=6), ileocystoplasty (n=6), commercial biomaterial (n=6), and University of Pennsylvania, Philadelphia, PA experimental biomaterial (n=6). A foley catheter was placed in all animals, while the ileocystoplasty and biomaterial groups also had a An experimentally naïve, 4-y-old male rhesus macaque (Macaca suprapubic catheter and penrose drain placed to ensure adequate mulatta) presented for weakness. On cage-side exam, the animal was urine drainage postoperatively. All groups received enrofloxacin bright and alert but was observed teeth-grinding with generalized postoperatively for the duration that the catheters were in place. The hindlimb weakness and ataxia. Sedated physical exam revealed no rate of UTI was 16.7% in the cystectomy group, 67% in the abnormalities. Whole-body radiographs were unremarkable and ileocystoplasty group, 33.3% in the commercial biomaterial group, point-of-care electrolyte and blood gas values were within normal and 33.3% in the experimental biomaterial group. The predominant limits. Cerebrospinal fluid was collected for fluid analysis and blood bacteria isolated were Enterococcus faecalis, Staphylococcus aureus, E. was collected for blood culture, complete blood count, and chemistry coli, Streptococcus dysgalactiae, Corynebacterium renale, and Aerococcus panel. The animal was given buprenorphine (0.005mg/kg) and sanguinicola. They were treated with appropriate antibiotics per meloxicam (0.2mg/kg) while diagnostic results were pending. The culture and sensitivity results. No urolithiasis was noted in any clinical signs progressed to include a left-sided head tilt, fine whole- group. This case highlights that UTI is the main complication of body tremor, and hindlimb paresis. Bloodwork was normal, blood bladder augmentation, and that postoperative antibiotics and culture was negative, and CSF analysis showed increased protein. continuous bladder drainage do not prevent infection. Gram + MRI showed multicentric and poorly demarcated hyperintense bacteria are the main pathogens, which could reflect elimination of lesions within the brain, brainstem, and cranial spinal cord. Given the Gram – bacteria by use of postoperative enrofloxacin. We discuss the clinical signs and diagnostic results, top differentials included viral incidence and significance of the isolated bacteria in human and or autoimmune etiologies. The animal was started on antiviral veterinary urinary tract infection. (galvancyclovir 13mg/kg), antiinflammatory (prednisone 1mg/kg), antimicrobial (ceftiofur 20mg/kg) , and gastroprotectant (famotidine PS38 Thyroid Follicular Carcinoma in an Adult Female Common 0.5mg/kg) therapy. Additional diagnostics included standard NHP Marmoset (Callithrix jacchus) serology panel (Herpes B, SRV, SIV, STLV), SV-40 serology, flavivirus serology panel (Dengue, West Nile, and Zika), crypococcus antigen JL Haupt*, SC Artim, MA Burns, JG Fox, S Muthupalani test, and toxoplasma PCR, all of which were negative. Cytomegalovirus (CMV) serology and PCR were positive. Given the Massachusetts Institute of Technology, Cambridge, MA positive CMV results, the lesions seen on imaging were consistent with acute disseminated encephalomyelitis, an autoimmune A 10-y-old female marmoset (Callithrix jacchus) presented with demyelinating condition that typically follows viral or bacterial cervical swelling and bruising during a quarantine examination. infection. CMV is a common beta-herpesvirus in nonhuman primate Further evaluation revealed poor dentition with significant gingival colonies. The virus undergoes periodic episodes of asymptomatic recession and pytalism. Diagnostics included CBC, serum chemistry, recrudescence but usually remains latent. Clinical CMV typically and pharyngeal culture. Ultrasound of the neck revealed bilateral requires immunosuppression, and in a research environment is enlargement of suspected cervical lymph nodes with intranodal commonly associated with infectious disease or transplant studies. hemorrhage and subcutaneous edema. No significant findings were This is a unique presentation of CMV since the animal was noted on bloodwork, with no pathogens identified on pharyngeal experimentally naïve and otherwise clinically healthy until the time culture. The animal was maintained on meloxicam and orbifloxacin of presentation, and highlights the importance of considering this to treat suspected reactive lymphadenopathy. Re-evaluation 1 wk virus as a differential in such cases. later revealed resolution of bruising, no change in ultrasonographic size of presumptive cervical lymph nodes, and bilateral soft tissue PS40 Increased Placental Fluid as a Gestational Complication in an opacity enlargement in the neck and tracheal deviation to the right African Green Monkey on radiographs. Aspiration of the node obtained hemorrhagic fluid consistent with a resolving hematoma and biopsy of the cervical skin TM Corey*1,2, R Pike1,2, DS Moddrelle3, M O’Connor1,2, XG Morton1,2, revealed no evidence of viral inclusions, neoplasia, or infectious A Chapwanya4, M Lawrence2,1 organisms. As no clinical signs were present, the animal was monitored for any enlargement of the cervical swelling or 1St. Kitts Biomedical Research Foundation, Basseterre, Saint Kitts and development in clinical signs. Approximately 4 mo after initial Nevis; 2RxGen, Inc., New Haven, CT; 3Medical Devices, WuXi evaluation, the animal developed dyspnea with stridor and open AppTec, Suzhou City, China; 4Theriogenology, Ross University mouth breathing following sedation for routine bloodwork. The School of Veterinary Medicine, Basseterre, Saint Kitts and Nevis animal was initially managed with antiinflammatories, but was recommended for euthanasia due to poor prognosis for recovery and A 7-y-old, 6.6 kg, female, uniparous African green monkey (AGM), possible compromise of overall colony health. Upon sedation for housed in an outdoor breeding enclosure, presented for routine terminal perfusion, the animal went into respiratory arrest. obstetric ultrasonographic imaging and was diagnosed with her Intubation was unsuccessful due to a large mass occluding the dorsal second pregnancy. Ultrasonography showed a fetus with a heart rate nasopharynx. On necropsy, a large yellow-brown cystic left thyroid of 160 beats per minute and a fetal crown-rump length of 16mm, mass extending dorsomedial to the larynx with associated lateral therefore estimated fetal age was 44 days. An intact amniotic sac deviation of the trachea was noted. Histologic findings were measuring 18mm in diameter was surrounded by echogenic fluid consistent with thyroid follicular carcinoma. Thyroid-related within the placenta, distending the placental lumen to 33 x 46 mm. pathology of marmosets is rarely reported in the literature, and to Other maternal physical examination findings were unremarkable, our knowledge this is the first report of thyroid follicular carcinoma apart from a distended abdomen disproportionate to gestational in a common marmoset. This case also highlights the need for duration. Follow-up ultrasound imaging performed 6 wk later anatomical description and imaging assessment of normal structures revealed a nonviable, partially disintegrated fetus with a crown- in the common marmoset. rump length of 25 mm, suggesting that the fetus died at

701 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 approximately 48 d of gestation. The amniotic sac was visible, and musculoskeletal coaching, and a daily movement maintenance the placenta remained distended with fluid. The endometrial lining program (MMP). The employees are trained to use specific posture, was thickened and irregularly shaped. The female was subsequently movement, and exercise tools to prevent musculoskeletal disorders treated with 5 units of IV oxytocin administered over 30 m, which (MSD) associated with manual material handling, awkward work failed to terminate the pregnancy. Seven weeks post fetal death, positions, and repetitive manual tasks in AST. Participation rates are hysterotomy was performed under general anesthesia to remove the high (>90%). All employees who requested individual coaching had placenta and dead fetus. The placenta was partially detached from excellent outcomes based on operational definitions (average of 94.8 the uterine wall and contained brown fluid and an intact amniotic pts out of 100) and the program has achieved longevity. The daily sac. Here we present an unusual gestational complication in MMP targets improved posture and body mechanics, maintaining nonhuman primates, which may represent a subchorionic flexibility in movements that offset the ergonomic risks in AST and hemorrhage or hydroallantois. In the case of gestational strategic strengthening to increase musculoskeletal resilience. In complications resulting in incomplete miscarriage in nonhuman addition, the program emphasizes training in protective work primates, medical options include administration of oxytocin, techniques and use of strategic micro-pausing techniques to combat misoprostol, or mifepristone as initial treatment. Due to the severity fatigue. Follow-up surveys demonstrate a universal benefit and of the fluid accumulation at the time of detection and the prolonged improved postural habits and awareness. There have been no duration of pregnancy postfetal death, a pharmacological musculoskeletal injuries, ROI is estimated to be 10.0 for the whole intervention was not further pursued, and surgery was performed to site with a potential cost avoidance of up to $102,059 for the evacuate the nonviable fetus and placenta immediately. department.

PS41 Making Sense of Workplace Injuries to Help Effect Change in PS43 Increasing Safety with Universal Chemical Containment the Workplace Levels

JR Krout*, S St. Omer, S Rasmussen G Repik*1, S Langston2

New York University Langone Health, New York, NY 1Environmental Health and Safety, Weill Cornell Medicine, New York, NY; 2Environmental Health and Safety, University of Florida, There are a multitude of ways staff can incur injuries while working Gainesville, FL in a research vivarium. Many workplace injuries can be prevented by proactively identifying jobs or tasks with a higher injury risk. Work- The administration of chemicals to research animals is continually related musculoskeletal disorders (WMSDs) are reported by the increasing and evolving. During the IACUC protocol review process, Occupational Health and Safety Administration (OSHA) as the “most IACUC reviewers, veterinary, and EHS staff are tasked with assessing widespread occupational health hazard facing our nation today.” hazards, including chemicals, in animal models. Assessments are WMSDs can occur due to lifting, standing in one position too long, often completed on a case-by-case basis, with inadequate data to repetitive movements, twisting, poor posture, and more. The creation reference, or by those with limited knowledge of the daily animal of a work environment where WMSDs can be avoided is optimal as facility operations. In addition, no regulatory guidance is provided this category of injury often takes a long time to heal, and in severe for handling chemical hazards in animals. This results in a lengthy cases, may require surgery resulting in lost workdays to turn into lost and inconsistent review. Consequently, the recommended safety work months. In line with the aims of our institution, we set out to procedures may be unintentionally inaccurate or incomplete, putting enhance our method of work injury reporting and tracking. both lab and animal care staff at risk. Recognizing a need for an Following modifications to our system of injury reporting we were improved review process, EHS and lab animal professionals from able to identify the type of injury, the anatomic location of the injury, multiple research institutions have come together to develop facility location where the injury occurred, the job task the employee chemical containment levels (CCL). Much like biosafety levels, CCLs was performing at the time of injury, and the type of equipment the will provide the foundation needed to provide an accurate risk employee was working with at the time of the injury. Based on injury assessment. As such, it is important to recognize that the CCL project reporting data, we determined 45.8% of the injuries reported from takes a “cradle to grave” approach, ensuring a review of all activities 2018 and 36.0% of injuries reported from the first half of 2019 could from administration to waste disposal. A list of the most commonly be categorized as WMSDs. Periodic workplace injury assessments used chemicals, administered in vivo, at participating institutions using data from the modified injury reporting system helped to was used as a starting point for this initiative. From there, guide staff ergonomic and safety training, and helped to identify collaborators evaluated the available toxicity and health hazard data areas for targeted ergonomic improvement. Over a period of 2 y, we for humans and specific animal models (if available). Classes and have seen a 38% reduction in the total number of injuries reported categories were assigned to the CCLs based on the severity and and a 34.8% reduction in the number of injuries categorized as commonality of the hazards. Overall, it was determined that the WMSDs. We highlight changes made to our system of injury majority of chemicals fit within 3 CCLs, with some chemicals needing reporting while using case-based examples to illustrate how we to be handled at a higher CCL for administration and a lower CCL obtained, analyzed, and used injury reporting data to help enhance for animal housing. Minimum housing standards (including workplace safety. engineering controls, PPE, etc.) for each containment level were determined. Information on comparable substitutions for these PS42 Benefits of Incorporating an Ergonomic and Injury Prevention controls was included to make CCL universally applicable and allow Program for Animal Facility Staff for flexibility among institutions with different resources. Our hope is that lab animal, IAUCU, veterinary, and EHS professionals can take J Harris2, E Straley*1, K Wingate1, J Duffy Rath3, W Rath3 the information provided back to their institution to help streamline their review process and improve safety for those working with 1Animal Sciences and Technologies, AstraZeneca, Gaithersburg, animals exposed to hazardous chemicals. MD; 2Gaithersburg Safety, Health, and Environment, AstraZeneca, Gaithersburg, MD; 3Duffy-Rath Physical Therapy, P.C., Barnegat PS44 To the Elevator and Beyond: Improving Work-related Light, NJ Conversations with Family and Friends as a Tool to Reduce Compassion Fatigue We implemented a collaborative and progressive ergonomic and injury prevention program June 2017. The program is based on a J Jones*, PA Lester musculoskeletal wellness and injury prevention blueprint which was customized to the tasks and personnel in the department. This Unit for Laboratory Animal Medicine, University of Michigan, Ann includes ergonomic assessments with corrective actions, a Arbor, MI progression of interactive training workshops, individual employee 702 Abstracts of scientific papers 2019 AALAS National Meeting

Our institution is consistently exploring novel methods to identify and improve communication regarding compassion fatigue. We Hamsters are a preferred species for monoclonal antibody (mAb) performed a survey over a 2-yperiod which indicated the majority of generation. The majority of hamsters used in research consist of respondents (64% and 63%, respectively) experienced symptoms of Syrian hamsters (Mesocricetus auratus) and as a result, most literature compassion fatigue. Recognizing the potentially detrimental and published procedural techniques are based around Syrian symptoms related to compassion fatigue, our department created the hamster anatomy and temperament. This posed a unique challenge Compassion Awareness Project, which works internally to prevent, when researchers at our facility requested the use of Armenian reduce, and mitigate the negative effects of compassion fatigue on hamsters (Cricetulus migratorius). Incorporating Armenian hamsters laboratory animal personnel. Although the initial programs covered a at our facility required our veterinarian, training manager, and variety of strategies aimed at reducing compassion fatigue, including technicians to learn the differences between Armenian and Syrian seminars on self-care, lectures featuring research performed at the hamsters to provide proper husbandry practices and immunization- university, and coworker support groups, these programs did not related techniques. Armenian hamsters have a more docile provide resources specifically targeting external social support temperament compared to their Syrian counterparts. This has networks. In addition, focus groups comprised of multiple laboratory allowed our training hamsters, breeding pairs, and weanlings to be animal personnel confirmed the feeling of unpreparedness when regularly pair or group housed. In addition, devices like scoops or talking about their work with people unfamiliar to laboratory animal bowls were deemed unnecessary for basic cage manipulations. Both medicine and indicated an interest in building skills to improve of these aspects resulted in a smoother husbandry workflow. They confidence. We felt these were important skills to improve as the lack have not shown any signs of aggression towards human handlers of understanding from the general public of what our profession and are instead relatively curious. Conversely, they are resistant to entails can hinder the ability to provide proper support to a lab manual restraint, and their excess loose skin, similar to other hamster animal care provider who may be experiencing compassion fatigue. species, further complicates their restraining. Veterinary treatments Having confirmed the need to develop and implement methods to and evaluations require patience and care from the handler due to improve communication confidence, we are designing a novel pilot the challenges associated with restraint. Any type of nontopical program to implement or strengthen preexisting communication veterinary intervention or immunization study work currently skills similar to communication styles typically associated with the requires general anesthesia to reduce as much stress to the animals as classic “elevator pitch.” The focus being creating an environment possible. Unlike for basic husbandry handling, we found using bowls where personnel can seek support from their external support system for topical treatments or tunnels for transferring to anesthesia without concern over judgement or lack of understanding of the machines provide the technician with greater control over the field. Judging from initial feedback from laboratory animal hamster while limiting potential escapes. Our veterinarian, training personnel, there is an increased sense of pride in their work and manager, and technicians keep up-to-date on current literature and improved support from family and friends. As a result, we plan to emerging techniques to provide the proper care to this infrequently continue expansion and development of confidence communication used and little known species. This will become even more necessary skills in our Compassion Awareness Project. as we continue to increase our breeding operation to secure longterm continuous access to this important animal model. PS45 Monkey Kitchen: Creative Food Enrichment for Nonhuman Primates PS47 Withdrawn

LT Alexander1, VA Davis*2 PS48 Development of Greek Tortoises and Hyalomma aegyptium Tick Feeding Model at BSL-4 to Study the Transmission 1Division of Laboratory Animal Resources, University of Kentucky, of Crimean-Congo Hemorrhagic Fever Virus Lexington, KY; 2Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY ES Lee*1, DA Bente2

Environmental enrichment is a critical part of maintaining animals in 1Animal Resources Center, University of Texas Medical Branch, a research setting, providing an outlet for species-specific behaviors League City, TX; 2Microbiology & Immunology, University of Texas and enhancing the psychological wellbeing of laboratory animals. Medical Branch, Galveston, TX Using food as enrichment items for nonhuman primates (NHPs) can be challenging, as these intelligent and curious animals crave novelty Crimean-Congo hemorrhagic fever (CCHF) is a tick- and interaction, while also having specific nutritional requirements. borne Nairovirus causing hemorrhagic fever in humans, and is a We have a standard of many healthy food enrichment offerings as select agent that must be maintained at the biosafety level-4 (BSL-4). well as nonedible environmental enrichment. However, a group of Latest evidence suggests that in addition, mammals and reptiles recently received NHPs did not show much interest in the usual and their ticks, specifically tortoises and Hyalomma aegyptium, play enrichment activities our department has used in the past. This troop a role in the maintenance and transmission of the CCHF virus appeared indifferent to picture books and television and showed (CCHFV) in nature. Greek tortoises (Testuda graeca) are native agitation and fear towards bubble and light machines. We decided to to the Mediterranean, and we have shown that they undergo get more creative with our edible enrichment leading to many novel subclinical infection with CCHFV but transmit the virus to their ways of presenting food. This has made an imaginative environment ticks. Researchers at our institution sought to characterize the role for technicians who get to come up with innovative ways of of Greek tortoises and tortoise-specific ticks, Hyalomma aegyptium, presenting food enrichments, thereby increasing staff morale and job as reservoirs and vectors, respectively, for CCHFV. Unique housing satisfaction, as reported via staff surveys. Video recordings of the and husbandry challenges of this model include that tortoises are NHPs have shown that these complex, fun, and fresh enrichment endothermic with specific temperature, humidity, UV light, and products have increased NHP inquisitiveness and activity (as behavioral requirements. For this project, the tortoise housing must measured by percentage of time spent interacting with the contain, and aid with tracking of, free-feeding ticks infected with enrichment items). CCHFV. Finally, the housing and husbandry procedures must satisfy the risk assessments of regulatory bodies such as the Center for PS46 Optimizing Husbandry Practices for Armenian Hamsters Disease Control and Prevention, and be safe for work within the (Cricetulus migratorius) full-suit BSL-4. Despite these challenges, we successfully created appropriate housing by modifying standard guinea pig tubs and M Short*, A Updike, D Calantropio-Covington, M Lu, A Napolitano, bedding, and adding species-specific hides, heat/UV lamps, and B Rendon, K Walton shallow water dishes. We adapted current tick-containment protocols for use with our tortoise housing. Here we share the husbandry and Bristol-Myers Squibb, Redwood City, CA housing procedures we developed for the care of Greek tortoises, and their ticks, within high biocontainment. 703 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

PS49 Housing Peafowl for Behavioral Studies of Hearing exposed to a 10% v/min CO2 flow rate based on prolonged durations of conscious dyspnea and ataxia. We conclude that emotional L Brossia-Root*1, J Cumming2, HE Heffner2 contagion may contribute to distress in cagemates at 10% v/min flow rate based on exacerbated jumping behavior. We propose that 30% 1 Department of Laboratory Animal Resources, University of Toledo, v/min CO2 should be used for euthanasia of grouped mice, and that Toledo, OH; 2Psychology, University of Toledo, Toledo, OH 50% v/min should also be considered humane.

Comparative studies of animals may involve species not commonly PS51 Nitric Oxide Inhibition Enhances Immunity of Neonatal Mice used in laboratories. One such study is determining the ability of To E. Coli-induced Meningitis in an Il-1 Dependent Manner birds to hear low-frequency sounds that are inaudible to humans (i.e., infrasound). Although pigeons and chickens hear infrasound, C Chambers*, J Skyberg mallard ducks and budgerigars, on the other hand, do not. Thus, the question arises as to which birds hear infrasound and which do not. University of Missouri, Columbia, MO Because a peacock’s train produces infrasound that is thought to be used in social signaling, we wanted to determine the hearing ability Neonatal meningitis-associated Escherichia coli (NMEC) is the leading of peafowl. This requires behavioral testing in which food is used cause of bacterial meningitis in premature infants. The neonatal to train them to report when they hear a tone, with the results used immune system has several differences compared to that of adults. It to construct an audiogram. One male and two female peafowl were is known that cord blood monocytes secrete decreased levels of obtained from a local farm. They are housed together in a large room mature IL-1 compared to adult monocytes in response to stimulation within an 11.3 m2 area enclosed with snow fencing attached to a with bacterial products, and that newborn humans and mice display sealed wood frame (2.1 m high) with a 2x4 wood perch (0.5 m high). increased nitric oxide (NO) levels compared to adults. We therefore The floor was originally covered with straw bedding which was wanted to determine the role of IL-1 and NO in the outcome of replaced with aspen bedding (10-15 cm deep) when it appeared that NMEC infection. In vitro, we show here that induction of IL-1 by dust from the straw contributed to a sinus impaction which required macrophages and microglial cells infected with NMEC is dependent surgical intervention in one animal. The animals were treated on NLRP3, a sensor molecule involved in the maturation of IL-1. We for parasites, and their nails inspected monthly and trimmed as also show here that adult IL-1 receptor knockout (IL-1R-/-) mice have necessary. They receive ad libitum water and are fed Purina Layena reduced survival and higher bacterial loads in the brain at 18 h crumbles, which they work for during the week in an auditory test following intracranial infection compared to wildtype, C57BL/6J chamber; on weekends, the peafowl are provided free access to the mice (n=15-21/group), indicating a protective role of IL-1 during E. food for 40 min each day. The animals are weighed daily while on coli meningitis. To confirm our results in a neonatal mouse model, we test, and this feeding regimen allows them to maintain a healthy infected 3-d-old C57BL/6J mouse pups intraperitoneally with NMEC body weight. This method of laboratory housing has maintained and simultaneously treated them with either anti-IL-1R antibody, or peafowl health while obtaining useable information about their an isotype control. Surprisingly, we did not see a significant effect of hearing abilities throughout the testing period. anti-IL-1R treatment with regards to bacterial loads in either the blood or brains of pups (n=6-8/group). It has been previously shown PS50 The Role of Emotional Contagion in Distress Exhibited by that nitric oxide can suppress NLRP3 activation and IL-1 production,

Grouped Mice Exposed to CO2 and that inhibition of nitric oxide enhances resistance to NMEC infection. Confirming previous results, we show here that treatment AD Moffitt*1, L Brignolo1, A Ardeshir2, M Creamer-Hente3 of pups with an inducible nitric oxide synthase (iNOS) inhibitor at the time of NMEC infection significantly decreased bacterial loads 1Campus Veterinary Services, University of California-Davis, Davis, compared to controls (n=5/group). Interestingly, we found that the CA; 2California National Primate Research Center, University of protective effect of iNOS inhibition was lost when pups were also California-Davis, Davis, CA; 3Comparative Medicine & Quality, The treated with anti-IL-1R antibody (n=11/group). This suggests that Jackson Laboratory, Sacramento, CA the protective effect of nitric oxide inhibition during NMEC infection is due to improved IL-1 signaling, and may indicate a target for The 2013 AVMA Guidelines for the Euthanasia of Animals recommends future therapeutics. a chamber volume displacement rate of 10% to 30% per minute (v/ min) when euthanizing small laboratory rodents with CO2. Group PS52 TKO hu-PBMC Humanized Mouse Model for HIV Research euthanasia of mice is a common practice, and grouping strangers is often avoided to minimize distress; however, emotional contagion, L Holguin*1,2, J Burnett2 which occurs between familiar animals but not strangers, has not 1 been studied in the context of group CO2 euthanasia. This study Center of Comparative Medicine, City of Hope , Monrovia, examined cagemate- and stranger-grouped mice exposed to 10%, CA; 2Center for Gene Therapy, Beckman Research Institute at the

30%, or 50% v/min CO2 to determine whether emotional contagion City of Hope, Duarte, CA plays a role in this context and whether that role is influenced by

CO2 flow rate. Videos of adult male C57BL/6J mice exposed to Currently there is no cure or preventative vaccine for HIV infection, different CO2 flow rates were scored for durations of dyspnea, ataxia, thus continued research is needed to end the HIV pandemic. While and consciousness as well as the numbers of face pawing (pain there are many animal models used in HIV research, none is used response) and jump (escape attempt) behaviors. Blood was collected more than the humanized mouse model. Humanized mouse models at time of unconsciousness and assayed for ACTH. Cagemates are all based around the use of immunodeficient mouse strains, most experienced significantly longer durations of conscious dyspnea and notably the NSG strain. While there are many uses for these current ataxia with 10% v/min CO2 compared with 30% and 50% v/min. models, they all have a major limitation in common: development of Similarly, strangers experienced significantly longer duration of graft-versus-host disease (GVHD). GVHD not only introduces conscious dyspnea with 10% v/min CO2 compared with 30% and variabilities into the research data, it leads to animal welfare 50% v/min and significantly longer duration of ataxia with 10% concerns. A new mouse strain, B6.129S-Rag2tm1Fwa Cd47tm1Fpl compared with 50% v/min. Cagemates showed significantly more Il2rgtm1Wjl/J (TKO), has been used to develop a humanized mouse jumps with 10% v/min CO2 compared with 30% and 50% v/min, model (TKO hu-BLT) that is resistant to GVHD development. We whereas jumping was unaffected by CO2 flow rate in strangers. At used TKO mice to develop a new hu-PBMC mouse model. Female 10% flow rate, cagemates showed significantly longer durations of TKO mice (n=7) were transplanted with human peripheral blood conscious dyspnea and ataxia, and significantly more jumps mononuclear cells (PBMCs) then monitored for engraftment by FACS compared with strangers. There were no significant differences in analysis. A cohort of these mice (n=3) were infected with HIV-1 and face pawing or ACTH levels between groups. We conclude that more monitored for plasma HIV viremia and CD4 T cell depletion. GVHD potential for distress exists when cagemate and stranger mice are 704 Abstracts of scientific papers 2019 AALAS National Meeting

development was monitored by clinical signs. As a control, NSG mice mice, we previously published that Hem-1 is critical for normal (n=6) were used to compare the TKO mice because the NSG hu- lymphopoiesis and innate immunity. However, the cell autonomous PBMC model is the most commonly used model. First, TKO mice functions of Hem-1 in individual immune cell types remain an transplanted human PBMCs supported engraft of human immune enigma. The objective of this study was to test our hypothesis that cells: CD3, CD4, and CD8 T cells as shown by FACS analysis. Second, Hem-1 is critical for migration and phagocytosis by myeloid cells and the TKO hu-PBMC model supports HIV-1 infection as seen by robust antiviral immunity, in a cell-autonomous manner. Our approach was plasma HIV viremia and depletion of CD4 T cells overtime as seen in to create conditional Nckap1lfl/fl mice, which were bred to LyzMCre the humans. Lastly, TKO mice showed a delayed onset of GVHD mice to delete Hem-1 specifically in myeloid cells. Using flow clinical signs (~14 d) compared to NSG mice. Based on these results, cytometry, fluorescence microscope, and time-lapse video the TKO hu-PBMC mouse model not only supports humanization microscopy, we found that neutrophils from Nckap1lfl/flLyzMCre mice and HIV-1 infection, but is also resistant to GVHD development exhibited defective F-actin polymerization and impaired migration in making this model valuable tool in HIV research. response to the chemoattractant 1 µM fMLP or LTB4, relative to LyzMCre control mice (n=4-5 male and female mice/group). Using PS53 Granulocyte Colony Stimulating Factor Plays a Role in the B 1µm fluorochrome labeled beads and flow cytometry, we found Cell Depletion of Bone Marrow in MNV-infected Stat1 Knockout that Nckap1lfl/flLyzMCre macrophages were unable to efficiently Mice phagocytose the beads (n=5 mice/group). Nckap1lfl/flLyzMCre mice were much more susceptible to 10 PFU influenza virus (H1N1/PR8) D Eldridge*, K Mears, C Hsu infection based on significantly increased body weight loss, increased peribronchial inflammation, reduced neutrophil and interstitial Comparative Medicine, University of Washington, Seattle, WA macrophage numbers, and increased pro-inflammatory cytokines in bronchoalveolar lavage fluid, 6 days post-oropharyngeal influenza Murine norovirus (MNV) is highly prevalent in laboratory mice and infection relative to LyzMCre control mice (n=5 mice/group). has been reported to infect macrophages, dendritic cells, T cells, and Collectively, our results reveal previously uncharacterized cell B cells. We previously showed that MNV infection in Stat1 knockout autonomous roles for Hem-1 in primary myeloid cells, and suggest (KO) mice (129S6/SvEv-Stat1tm1Rds) caused a significant depletion of that Hem-1 is critically important for effective antiviral immunity. developing B cell populations in the bone marrow (BM). Concurrent Statistical significance was determined by a two-tailed student’s with this B cell depletion, MNV infected Stat1 KO mice also had a t-test. significant increase in BM granulocytes and serum granulocyte colony stimulating factor (GCSF). Therefore, we hypothesized that PS55 Conditional Deletion of Nckap1l Encoding Hematopoietic the increased GCSF and BM granulopoiesis directly contributed to Protein-1 Alters B Cell Maturation, Antibody Production, and the BM B cell depletion observed in MNV infected Stat1 KO mice. To Immune Responses to Influenza Virus test this hypothesis, uninfected female 5-9-wk-old Stat1 KO mice (n=5 per group) were IP administered either isotype IgG (10 μg/ AV Avalos*, H Park, N Suwankitwat, T Iwata, B Iritani mouse) or GCSF (0.5 μg/mouse) daily for 7 days in order to induce granulopoiesis. BM B cells were then evaluated by flow cytometry to Comparative Medicine, University of Washington, Seattle, WA determine if B cell losses were observed, similar to that seen in MNV infected mice. We show that daily IP administration of GCSF for 7 Mutations in genes encoding actin-regulatory proteins can lead to days caused a significant depletion (P < 0.05) of pre-B/immature and primary immunodeficiency diseases (PIDs) characterized by mature B cells (2.2 fold and 2.4 fold, respectively) in the BM, as well impaired immunity and reoccurring debilitating infections. Utilizing as a concurrent increase of granulocytes (2.6 fold, P < 0.05) as a chemical mutagenesis strategy in mice to discover novel genes expected. To further evaluate the role of GCSF, we administered involved in immune system function, we previously identified a either anti-GCSF antibody (10 μg/mouse) or isotype IgG (10 μg/ strain,NTB.1, with reduced B and T cell numbers due to a non-coding mouse) daily for 7 days to MNV infected Stat1 KO mice (n=5 per mutation in the Nck-associated protein 1-like gene encoding group) to determine whether preventing granulopoiesis in the BM Hematopoietic protein-1 (Hem-1), a hematopoietic cell specific actin would rescue the B cell losses observed after infection. We show that regulatory protein. NTB.1 mice are severely immunodeficient, daily IP injection of anti-GCSF antibody for 7 days postinfection in characterized by reduced B and T cells, impaired neutrophil Stat1 KO mice resulted in increased (although not statistically migration and phagocytic abilities. To determine why B cells are significant) pre-B/immature and mature B cell populations (1.5 fold reduced in NTB.1 mice, we utilized the Cre-LoxP system to generate and 1.7 fold, respectively) and decreased granulocyte populations mice (Nckap1lfl/flMb1Cre) that were conditionally deleted (1.4 fold) in BM compared to MNV infected mice. These results for Nckapl1 in a B cell specific manner. We hypothesized that Hem-1 suggest that granulopoiesis induced by GCSF plays a role in the BM is important in B cell development, B cell migration, and antibody B cell losses seen after MNV infection in the absence of Stat1. Further production. Flow cytometry revealed that deletion of Hem-1 resulted study is warranted to determine why MNV infection induces in impaired follicular and marginal zone B cell development and increased GCSF and granulopoiesis in Stat1 KO mice. reduced innate-like B1a cells (4 independent experiments with n≥4 group). Using flow cytometry and ELISA, we found that Nckap1lfl/ PS54 Conditional Disruption of Hematopoietic Protein-1 In Mice fl Mb1Cre mice had decreased ability to form germinal centers in Reveals an Essential Role for Hem-1 In Myeloid Cell Functions mediastinal lymph nodes and impaired antigen-specific antibody production 10 days following oral pharyngeal influenza PR-8 N Suwankitwat*1, H Park1, T Iwata1, S Libby2, D Liggitt1, B Iritani1 infection, relative to WT littermate mice (n=6 per group). Nckap1lfl/ fl Mb1Cre had reduced antibody production 5 days following 1Comparative Medicine, University of Washington, Seattle, immunization with heat-killed S. pneumonia (a T-independent WA; 2Laboratory Medicine, University of Washington, Seattle, WA antigen), and reduced long-lived antibody production at 3 and 4 wks post-immunization with the T-dependent immunogen Keyhole Hematopoietic protein1 (Hem-1) is a hematopoietic cell-specific Limpet Hemocyanin (KLH) (n=6 per group). These results suggest subunit of WAVE (WASP-family verprolin homologous protein) that Hem-1 is essential for normal B cell development and antibody complex, which acts downstream immune receptors (including BCR, production in a B cell specific manner. We predict that mutations TCR, TLR, and cytokine receptors) to stimulate filamentous actin in Nckap1l could be the cause of some undefined PIDs in humans (F-actin) polymerization. Inactivating mutations in NCKAP1L, the characterized by deficient antibody production and increased gene encoding Hem-1, have been recently associated with primary susceptibility to bacterial or viral infections. immunodeficiency disease in humans, and high NCKAP1L expression has been associated with a poor prognosis in chronic lymphocytic leukemia. Using constitutive Nckap1l null

705 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

NG supplementation. Weights were monitored daily and mice were PS56 Inhibition of Oxidative Phosphorylation but not Glycolysis removed at 30% loss of initial weight. There was a significant Attenuates Lung Injury Caused by H1N1 Influenza A Virus difference (P < 0.05) seen in the maximum percent weight loss of Infection mice given NG in all viral doses used. There was significantly decreased mortality (P < 0.05) seen in the mice given NG at the K Nolan*1, L Baer2, A Nelson2, K Stanford2, L Doolittle2, L Rosas2, I middle and high viral doses. Flow cytometry on cells from Davis2 bronchoalveolar lavage fluid, collected at 6 d postinfluenza infection from female mice (n=10) given the middle dose, demonstrated no 1ULAR, Ohio State University, Columbus, OH; 2Ohio State significant differences (P > 0.05) in percentage and absolute numbers University, Columbus, OH of inflammatory cell populations (specifically resident and interstitial macrophages, neutrophils, eosinophils, T-cells, and B-cells). In Despite availability of vaccines and antiviral drugs, seasonal summary, the results of this study show that supplementation of NG influenza A virus (IAV) epidemics cause >300,000 deaths/year can be beneficial in reducing weight loss and mortality in mice worldwide. IAV infection alters lung epithelial cell metabolism. This infected with various doses of mouse-adapted influenza virus. promotes a shift towards glycolysis and away from oxidative phosphorylation (OXPHOS) for ATP production. We hypothesized PS58 Evidence of Gastrointestinal Microbiome Stability with that this shift benefits the virus rather than the host and that Chronic Simian Immunodeficiency Virus Infection in Sooty inhibition of glycolysis would improve infection outcomes. C57BL/6 Mangabeys (Cercocebus atys) mice (n=5-6/group) were intranasally inoculated with mouse influenza A/WSN/33 (H1N1). Controls were mock-infected with R Bochart*1, GK Tharp2, M Crane1, BV Madeti2, A Ericsen2, S Jean1, J virus diluent. To inhibit glycolysis, mice were treated daily from 1 Cohen1, S Bosinger2 day postinfection (dpi) with 1g/kg 2-deoxy-D-glucose (2-DG). To block OXPHOS, mice were injected every other day from 1 dpi with 1Division of Animal Resources, Yerkes National Primate Research 0.8 mg/kg rotenone (ROT). Treatment controls were treated with Center, Emory University, Atlanta, GA; 2Division of Microbiology & saline. Carotid arterial oxygen saturation (SaO2) was measured using Immunology, Yerkes National Primate Research Center, Emory the MouseOx system at 2, 4, and 6 d postinoculation (dpi). Open University, Atlanta, GA circuit calorimetry and measurement of mouse activity were performed simultaneously using the Oxymax/CLAMS metabolic Human immunodeficiency virus (HIV) remains a global concern chambers from 5-6 dpi. On d 6, mice were euthanized and lungs with approximately 36.9 million people living with the virus and harvested. Viral replication was quantified by serial dilution and 940,000 HIV-associated deaths each year. The sooty mangabey (SM) plaque assay of lung homogenates on MDCK cells. Whole lung is a natural host of simian immunodeficiency virus (SIV) as they wet:dry weight ratios were calculated as an index of intrapulmonary maintain high viral loads and avoid progression to acquired immune fluid accumulation. Relative to controls, IAV infection induced severe disease syndrome (AIDS), which is unlike HIV in humans, and SIV in hypoxemia and pulmonary edema at 6 dpi. There was a significant rhesus macaques (RhMs). Previous studies at Yerkes National decline in nocturnal activity and a decrease in the respiratory Primate Research Center (YNPRC) have identified that a primary exchange ratio (RER), indicating a shift towards increased lipid tissue site differentiating the pathogenic versus non-pathogenic catabolism for ATP generation. Treatment of IAV-infected mice with outcome in SIV infection is the gastrointestinal mucosa. This work the glycolysis inhibitor 2-DG and the OXPHOS inhibitor ROT did not demonstrated that unlike non-natural host species (e.g. RhMs), alter lung IAV titers; however, 2-DG significantly worsened IAV- natural host species retain their mucosal integrity, do not exhibit induced hypoxemia and further decreased nocturnal activity. In luminal microbiota translocation, and harbor a toll-like receptor-4 contrast, ROT treatment restored SaO2 to normal levels, normalized gene mutation which dampens mucosal inflammatory responses to RER, and significantly attenuated IAV-induced pulmonary edema. gram-negative bacteria. Collectively, these data demonstrate that the Blockade of OXPHOS with ROT improves outcomes in IAV-infected gastrointestinal environment and mucosal immunity play a strong mice while inhibition of glycolysis exacerbates severity. This role in determining disease outcome. In this study we characterized indicates that a shift to glycolysis is protective in influenza and the gastrointestinal microbiota of the SM and its relationship to SIV suggests that OXPHOS may be a therapeutic target in this disease. status in comparison with RhMs. In comparison to the non-natural host, we hypothesized the natural host would show evidence of a PS57 Nutritional Gel Supplementation Minimizes Weight Loss in beneficial shift of microbiota with SIV infection allowing them to Mice Infected with Influenza A/PR/8/34 Virus prevent disease progression. Fecal samples from fifty SMs and thirty RhMs of mixed SIV positive and negative controls housed at YNPRC J Felgenhauer*, J Brune, T Brabb, C Frevert were collected and 16S ribosome-based taxonomic characterization was performed by amplification of the V4 hypervariable region. With Comparative Medicine, University of Washington, Seattle, WA chronic SIV infection in SMs, no statistically relevant changes were found (p < 0.05) for Shannon diversity, or at taxonomic levels that Influenza places a large burden on public health, and is therefore a have been previously reported to change with pathogenic SIV highly researched disease process that uses in vivo studies regularly infections. Our data was consistent with previous literature, showing with mice as a commonly used model. A complication of the clinical some statistical differences primarily at the genera level with SIV course of influenza infection in mice is anorexia and dehydration infection in RhMs. In contrast to pathogenic SIV infections, we leading to excessive weight loss often resulting in the early removal conclude SMs with chronic SIV infection revealed microbial stability of mice from study based on euthanasia criteria. To reduce the of potentially pathogenic taxonomic genera that have been shown to number of mice prematurely removed from an experiment, we modulate mucosal inflammation and immune activation, which may assessed the use of a nutritional gel (NG) supplement that is further potentiate disease progression. palatable and provides hydration, calories/carbohydrates, and electrolytes. We hypothesized that when compared to the standard of PS59 Gut Microbiota Alterations in Marmoset Wasting Syndrome: care, supplementation of NG to mice infected with influenza would A Cross-Population Study lead to decreased weight loss and mortality without impacting the immunologic data of a study. Both male and female C57Bl6/J mice were infected with mouse-adapted influenza A/PR/8/34 virus at RE Cooper*1, L Mangus1, J Wright2, R Lamendella2, J Mankowski1 low (0.2 times the lethal dose 50 (LD50); SOC n=21, NG n=20), medium (0.5 LD50; SOC n=30, NG n=27), or high doses (1.25 LD50; 1Molecular and Comparative Pathobiology, Johns Hopkins SOC n=23, NG n=22). Mice were provided with either the standard University, Baltimore, MD; 2Wright Labs, LLC, Huntingdon, PA of care (moistened pellets + hydration gel), or moistened pellets +

706 Abstracts of scientific papers 2019 AALAS National Meeting

In captivity, common marmosets (Callithrix jacchus) are susceptible to marmoset wasting syndrome (MWS), characterized by chronic PS61 Wound Management of Foreign Body Reactions in Nerve- enteritis and progressive weight loss. MWS, like human grafted Dorset Sheep (Ovis aries) inflammatory bowel diseases, may be associated with gut microbial dysbiosis; thus, investigating the gut microbiome of marmosets may W Hanson*1, U Blas-Machado2, C Roach1 inform prevention and management of MWS. Using 16S rRNA sequencing of rectal swab samples, we characterized the gut bacterial 1Division of Animal Resources, Emory University, Atlanta, microbiota of marmosets in captive colonies with differing MWS GA; 2Department of Pathology, University of Georgia, Athens, GA prevalence: German research university (MWS 0%; healthy n=15), German primate center (MWS 0%; healthy n=18), and United States Two of seven 2-year-old, female Dorset sheep (Ovis aries) on a nerve (US) research university (MWS 5-10%; healthy n=18; MWS-affected regeneration study received 7-cm-long bioengineered grafts imbibed n=6). German-origin (GO) marmoset samples were collected at three with autologous stem cells in their median (left forelimb) and sural time points: 1) upon arrival at the US facility (baseline), 2) following (right hindlimb) nerves. Postoperative complications, specifically transition to US diet in quarantine (100 d), and 3) following social surgical site edema and dehiscence, occurred on d 6 in the forelimbs integration with US colony marmosets (1 y), to dissect the of both animals. The edema and inflammation in the first sheep contributions of diet and social integration in shaping the gut developed into an abscess (cultured as Trueperella pyogenes, microbiota. At baseline, GO samples displayed greater phylogenetic Fusobacterium necrophorum, and Bacteroides fragilis) despite medical diversity and less species evenness, features of greater microbial treatment, and a draining tract eventually resulted. Daily treatments resilience, compared to healthy US samples. Evenness increased and included standard wound management, analgesia, and multiple richness decreased in GO samples at 100 d, but rebounded to baseline courses of antibiotics. The dehiscence in the second sheep was treated at 1 y. Beta analysis revealed increasing similarity of US and GO gut with standard wound management (including wet-to-dry and microbial communities at 1 y compared to baseline; a greater shift in hydrogel bandages), analgesia, and preemptive antibiotics. Despite GO samples suggested a role of diet in shaping the gut multiple attempts to debride and close the wounds, both microbiota. Peptostreptococcaceae, implicated in colorectal cancer and complications progressed until a discrete, long, thick, white, fibrous colitis in humans, was present in 14/18 (78%) US samples and 4/33 tissue was found protruding from the sites. This tissue was sampled (12%) GO samples at baseline. Both increased occurrence (11/29; for histology, which revealed a granulomatous inflammation 38%) and relative abundance were found in GO samples at 100 d; consistent with a foreign body reaction to an undetermined, further enrichment (17/30; 57%) occurred at 1 y, implicating diet and eosinophilic, spheroidal material. Both animals were taken to surgery possibly integration as causative. When comparing healthy and for removal of the white, fibrous tissue (presumed to be the MWS-affected US animals, Prevotella, associated with inflammation bioengineered conduit) as well as debridement of necrotic tissue. in mice and humans, was significantly enriched in the MWS-affected Both sheep achieved complete resolution of their forelimb group. Significant differences in the microbiota of GO versus US complications within 4 weeks post-debridement. Throughout this marmosets suggest that the gut microbiome plays a role in time, the sheep maintained appropriate mentation, activity, appetite, development of MWS. Enrichment of Prevotella in animals with MWS hydration status, and pain management. Despite the pronounced provides a compelling direction for future studies of disease foreign body reaction witnessed in these two cases, all hindlimbs of pathogenesis. the seven sheep healed normally after the engraftment procedure. However, all forelimbs of the post-surgical forelimbs developed some PS60 Systemic Coccidiosis Causing Fulminant Mortality in a level of edema and inflammation, suggesting that forelimb anatomy Colony of Wild-caught European Starlings (Sturnus vulgaris) (i.e., high tension of skin, increased friction of incision site at axilla) contributes. In these two cases, it is proposed that two separate and JG Vilches-Moure*, SA Felt unrelated post-surgical events acted as inciting causes for a heightened inflammatory response, ultimately resulting in a foreign Comparative Medicine, Stanford University, Stanford, CA body reaction. Foreign body reactions should remain as a differential despite previous successful engraftments. Disseminated disease caused by coccidia (systemic isosporosis, also known an atoxoplasmosis) is a common parasitic disease of passerine PS62 Nursing Care of Rabbits with a Spinal Cord Injury birds. Twenty-one European starlings (Sturnus vulgaris) were caught in California in 2016. The time between arrival and onset of clinical M Nigro*, LS Bird signs ranged from 2 d to 2 mo, and signs included anorexia, dropping food, neurologic deficits, conjunctivitis, and sudden death. At the Comparative Medicine Resources, Rutgers University, Piscataway, NJ time of death, birds ranged from fledglings to adults (n=21), with most of the birds ranging from 3-6 wk of age (n=10). Hematology Spinal cord injury (SCI) surgery models and the related postoperative revealed peripheral eosinophilia and lymphocytic intracytoplasmic care is well established in rodents, however limited information is inclusions in most, but not all, birds. Fecal evaluation revealed cysts available for a rabbit model. Our veterinary staff was approached measuring 39 x 39 to 39 x 44 mm. Necropsy revealed decreased with clinical management of a surgical SCI protocol in rabbits. All pectoral musculature, blepharedema, prominent keels, enlarged rabbits received a transverse resection of the cord at T8-9; the pale livers and spleens, and thickened intestinal walls. Histology experimental group received a polymer implant around the spinal revealed florid lymphocytic inflammation with intracytoplasmic cord, providing a cylindrical space for cord repair and regeneration. parasitic inclusions commonly affecting liver, spleen, conjunctiva, We present the nursing challenges of the model and animal connective tissues surrounding the thyroid glands, and bursa of management. We created a postoperative care plan that addressed Fabricius. Samples collected for molecular screening of common major concerns related to SCI, such as ambulation, bladder atony, avian pathogens were negative. Oral ponazuril treatment was fecal incontinence, appetite, and postoperative analgesia. The plan initiated, and this improved the clinical symptoms in some, but included chewable probiotics and critical care diet gel for not all, birds. Because of the variation in clinical manifestations, gastrointestinal concerns, bandaging feet to prevent autophagia, and circulating parasitic load, intestinal parasitic burden, and parasitic bladder expression. Infant pants protected against urine scald and stages present, we query if these differences represent the true created an additional barrier to prevent autophagia. All rabbits were spectrum of lesions observed in infection with a single parasite, or if given a sustained release opioid and nonsteroidal antiinflammatory they represent infection with more than one (or different strains of) analgesic. A postoperative schedule was created to perform coccidian parasites. assessments and care every 8 h, which included manual bladder expression, pain assessment, cleaning of the perineum with pants change, bandage integrity check, and evaluation of food and water consumption. Food and water in shallow bowls, hay, and fresh fruits

707 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 and vegetables were provided on the cage floor. Hay was also did not reoccur. However, 1 male bird developed a severe beak available in stainless steel whisks, and fruit and vegetables were malformation resulting in inability to close the beak. This bird was hand fed multiple times a day. All rabbits handled the recovery euthanized, and histopathology of the beak showed extensive process differently, behaviorally and physically. Their level of necrosis and inflammation. Eosinophilic, intracytoplasmic inclusion mobility, mentation, and bladder tone varied regardless of their bodies were present in mononuclear cells in the nasal and oral group designation. Fine attention to detail was crucial in keeping the submucosa, which is consistent with poxvirus infection. One mo rabbits comfortable for the duration of the study. The success of the later, an additional cohort of cowbirds was brought indoors for over- experiment, ease of clinical management, and carefully designed wintering. Several of these birds developed similar small pox lesions nursing care indicates the viability of the rabbit as a spinal cord on the digits. All birds remained otherwise clinically healthy, and all injury model. lesions resolved spontaneously. Other than the 1 bird with beak necrosis, all pox lesions were cutaneous. Since no birds displayed PS63 Polyglactin 910 Suture-related Pseudoinfection in a Yucatan clinical signs until being moved to the indoor housing facility, Pig transport stress likely led to the development of the pox lesions.

DE Collins*, B Simons PS65 Feline Atypical Mycobacterial Panniculitis Caused by Mycobacterium porcinum Center for Comparative Medicine, Baylor College of Medicine, Houston , TX HR Holcombe*1, A Mannion1, A Sheh1, JG Fox1, T McCollester2

Cutaneous suture reactions related to inappropriate suture material 1Division of Comparative Medicine, MIT, Cambridge, MA; 2Private selection or delay in removal are well documented in both the human Practice, Rochester, MA and veterinary medical literature. However, foreign body extrusion of an internal absorbable suture has not been well described in the A 12-y-old castrated male DLH cat presented with multiple dry veterinary literature. A female, intact Yucatan pig arrived in apparent lacerations measuring 3mm to 1cm on the caudal abdomen. The cat good health, and due to closure of study funding, remained naive. As failed to respond to systemic amoxicillin/clavulanic acid and topical such, a routine ovariectomy was performed for animal health and as chlorhexidine soaks. Within 2 wk, additional lesions, some with required by the vendor to allow for potential adoption. purulent discharge, were present, and Achromobacter spp., sensitive to Polydioxanone was used for the entire procedure except for ovarian amoxicillin, was cultured. Lesions did not resolve after 2 mo of pedicle ligation in which Polyglactin 910 was elected due to its low antibiotic therapy, and complete surgical excision was attempted. memory, ease of handling, and knot security. Approximately 3-4 mo Histologic analysis showed multifocal inflammatory infiltrates later, recurrent small masses on bilateral flanks of the animal were associated with granulation tissue and small cavitated spaces with noted. Diagnostics including cytology, fungal, and bacterial culture, acid-fast-negative, Gram-positive bacilli bacteria, leading to a and bloodwork all supported a noninfectious etiology. Biopsy results presumptive diagnosis of Actinomyces spp. New abdominal lesions from the site indicated a suture-related pseudoinfection. Despite noted at suture removal continued to wax and wane but never treatment including topical, oral, and parenteral antibiotics, surgical resolve despite varied, intensive systemic and topical treatment. The debridement, and nonsteroidal antiinflammatory drugs, these lesions cat did not exhibit additional clinical signs until 18 mo after the initial did not resolve. Eventually, chronic lameness developed in the worse presentation when he became lethargic and anorexic. A fast-growing affected side, and euthanasia was elected. Postmortem necropsy mycobacterium was then cultured from the draining lesions and definitively demonstrated a suture-related pseudoinfection with submitted for identification by whole genome sequencing (WGS). extrusion of suture material from the ovarian pedicle ligatures The cat continued to deteriorate and was euthanized prior to final through the body wall and skin leading to numerous sterile abscesses identification of the isolate. The draft genome of the isolate was in the flank areas. Though well documented in the human literature, sequenced followed by assembly into contigs with SPAdes and gene this is the first report of Polyglactin 910 suture-related annotation with RAST hosted by PATRIC. The resulting genome was pseudoinfection in a veterinary patient. While this may be an isolated 6,796,350 bp in size with a GC content of 66.8% and contained 6,630 incident, it may also indicate an increased sensitivity by Yucatan pigs protein-coding genes, 52 tRNA genes, and 5 rRNA genes. Using the and support presurgical assessment with vicryl hypersensitivity tetra correlation search function hosted, the cat mycobacterium patch testing similar to what is performed in at risk human patients. genome was most closely related to Mycobacterium porcinum, a fast- growing mycobacterium that belong to the M. fortuitum third PS64 Avian Poxvirus Infection in a Colony of Brown-Headed biovariant complex. Whole-genome phylogenetic and average Cowbirds (Molothrus ater) nucleotide identity analyses confirmed the cat mycobacterium genome as M. porcinum. M. porcinum has not been reported as a cause CE Blevins*1, AK Brice1,2, R Duran-Struuck1,2 of feline atypical mycobacterial panniculitis, nor has it been reported as a zoonotic agent. Historically, inconsistencies regarding published 1Department of Pathobiology, University of Pennsylvania, mycobacterium genomes made speciation difficult prior to WGS. Philadelphia, PA; 2University Laboratory Animal Resources, Accurate genome sequencing and reporting is essential due to the University of Pennsylvania, Philadelphia, PA zoonotic potential of some mycobacterial spp.

A cohort of 24 adult male and female brown-headed cowbirds PS66 Observational Learning Facilitates Positive Reinforcement (Molothrus ater) were wild-caught using an outdoor funnel trap on an Training in Macaques (Macaca mulatta) IACUC-approved protocol. The birds were quarantined in an outdoor aviary for 2 mo prior to being brought indoors. Three days K Coleman*, L Houser following arrival to the indoor facility, 9 of 24 birds were reported for multiple, tan, proliferative, 2.0mm-3.0mm masses on 1 or more digits. Oregon National Primate Research Center, Beaverton, OR Additionally, 2 of the birds had a small nodule located on the dorsal maxilla. All birds otherwise appeared clinically healthy. One wk later, Studies have pointed to the many benefits of positive reinforcement a female bird developed 5 3.0mm-5.0mm masses on multiple digits. training (PRT) to animal wellbeing. However, despite the benefits, This bird was euthanized and submitted for postmortem training can involve a large initial input of time, which may preclude examination. Microscopically, there was severe multifocal its use in some facilities. Further, not every animal learns at the same lymphohistiocytic dermatitis with hyperkeratosis and intraepithelial rate. Inhibited or shy animals may take longer to train than eosinophilic intracytoplasmic inclusion bodies (Bollinger bodies), exploratory animals. Therefore, finding alternate training methods which are pathognomonic for avian poxvirus infection. Over the next that can make the process less time consuming could be of great month, masses on other affected birds resolved spontaneously and value. Because primates are known to learn by observing the actions

708 Abstracts of scientific papers 2019 AALAS National Meeting

of others, one potential alternate technique is to have subjects watch showed the macaques had a preference for yogurt covered treats conspecifics being trained. In this study, we examined whether (74.13 ± 21.55) followed by grapes (58.74 ± 20.01), dried pineapple observational learning facilitates training for shy female rhesus (33.13 ± 12.96), banana chips (25.73 ± 11.72), dried apricots (20.35 ± macaques (Macaca mulatta). All females were housed with their 5.08), and peanuts (19.99 ± 10.69). Individual preferences were infants. We first trained 6 bold monkeys (“demonstrators”) for 2 tasks confirmed by the reinforcer evaluations in which the primates were using PRT: touch a target and present for menses check. We then more likely to perform a task for their most preferred item (average trained 12 shy monkeys for the same tasks. Eight of these monkeys engagement score > 80%) versus the least preferred or control (“observers”) watched the demonstrators being trained, while the condition. When choice for all food items was examined over a others (“non-observers”) had not. As expected, bold monkeys were month there was a great deal of instability in preference as shown by more likely than shy monkeys to learn this task within 12 sessions less than significant Kendall’s tau correlation coefficients (P > .05). (P < 0.05). Observers were more likely than nonobservers to perform However, when stability of the most preferred item alone was the task as well. While 6 of the observers reliably presented, none of examined, the item chosen first remained consistent for most of the the nonobservers learned this task. To see if observational learning macaques (12 of 14). On average the MSWO required three trials to applied to the infants, we trained infants to touch a target. Nine of identify preference and took approximately 4 m to complete. These the infants were with their mothers when they were trained for the 2 results suggest the MSWO is a practical and effective method to tasks, 6 watched a female other than their mother being trained, and identify preferred food items and can be used to inform 5 did not observe any training. Infants whose mothers were trained reinforcement choices in PRT programs. learned this task sooner than other infants (P < 0.01). These results suggest that observational learning may facilitate PRT for inhibited PS69 Programmed Training-Primate Computer Learning as a monkeys. Further, it may be an effective tool for training young Model of Positive Reinforcement Training monkeys. K Morrisroe* PS67 How Can the IACUC Encourage and Expand Positive Reinforcement Training Programs? Behavioral Management, Washington National Primate Research Center, Lynnwood, WA M Bloomsmith*, JE Perlman Positive reinforcement is defined as adding a desired stimulus Yerkes National Primate Research Center, Atlanta, GA following a behavior, resulting in the behavior being more likely to occur in the future. Being able to program the exact timing and The process of reviewing and approving research protocols by the amount of rewards while maintaining absolute consistency makes IACUC can be a powerful method for enhancing positive computer-based training an ideal model of positive reinforcement as reinforcement training of monkeys assigned to research projects. well as a valuable tool for data collection. Automated systems can Promoting the increased use of positive reinforcement methods is continually calculate rates of correct trials, giving the trainer compliant with many statements in The Guide for the Care and Use of immediate feedback on the animal’s learning and the ability to make Laboratory Animals (2011). Many behaviors that are commonly part of adjustments that will maximize success. Such computer-based research procedures can be trained with positive approaches, training systems can be set up remotely, thereby allowing 1 person to including shifting to a new cage or location for testing, temporary train multiple animals at a time while also eliminating the potential separation from a partner, receiving an injection, performing a blood biases that come with human interaction. One of the major challenges withdrawal, or moving into a restraint chair. When reviewing to computer-based training is anticipating the successive IACUC protocols, IACUC members can ask questions about the approximations needed for learning. Clear and accurate training planned training processes, the expected duration of training prior to plans, which break down each task into its smallest components, initiating research procedures, criteria for considering a monkey to need to be developed by the trainer and communicated to the task be fully trained on a particular behavior, what will be done if a programmers in order to allow the animals a seamless learning monkey does not adequately learn the procedure on time, and how process. An accessible user interface which allows the trainer to move regression will be handled. To accomplish this, at least some IACUC the animal through each approximation with minimal interruption members must be familiar with animal training methods, and be can help create that kind of seamless experience. At our facility, aware of the types of behaviors that are currently being trained computer learning is used in a variety of ways, from environmental across the laboratory animal science field. Some facilities can refer enrichment in behavioral management to virtual reality testing in investigators to primate training specialists to help plan for animal neuroscience. This introduction will show the possibilities for training as studies are being designed and this process will be computer learning with examples of tasks that range from the described. Some IACUCs may include assessment of animal training beginning of touch screen interaction to complex memory tasks in procedures when they conduct postapproval monitoring of research virtual reality. studies. The IACUC can be instrumental in encouraging investigators to consider the role that positive reinforcement training can play in PS70 The Closed Box Chair as a Refinement in Nonhuman Primate their research, and in improving the welfare of research primates. Sperm Collection

PS68 Assessing Food Preference and Reinforcer Effectiveness in L Houser*, C Ramsey, F de Carvalho, B Kolwitz, K Coleman, C Hanna Laboratory Housed Cynomolgus Macaques (Macaca fascicularis) Oregon National Primate Research Center, Beaverton, OR AN Rehrig*1, L DiVincenti2, D Napolitano3, D McAdam1 Collecting semen from macaques for assisted reproductive 1University of Rochester, Rochester, NY; 2Seneca Park Zoo, Rochester, technology (ART) procedures is critical for the success of those NY; 3Hillside Children’s Center, Rochester, NY projects. In order to get the samples, monkeys are often restrained in open restraint chairs (ORC) using the pole and collar technique. Preference assessments systematically and objectively identify items While commonly used, this restraint is not tolerated by all monkeys; that are preferred and could serve as reinforcers for positive some become anxious or aggressive towards the poles and people. In reinforcement training (PRT). This study used a multiple stimulus an effort to refine this procedure and improve the welfare of the without replacement (MSWO) preference assessment to determine monkeys, we recently modified a closed box chair (CBC), a clear, preference hierarchies of six food items with Cynomolgus macaques plexiglass box in which the monkey is trained to sit. Unlike the ORC, (n=14; 11 males and 3 females). Additionally, seven macaques the CBC does not require pole and collar, and although legs are completed concurrent-schedule reinforcer evaluations to confirm the secured, the monkey’s arms and neck are not restrained, allowing preferred items efficacy as reinforcement. Average food ratings more freedom than with the ORC. Because it is thought to be an

709 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 improvement in welfare, use of CBCs has increased in recent years; in an effort to increase strength, dexterity, and flexibility. Veterinary however, there are few studies demonstrating its effects on scientific technicians rate each chimpanzee on 6 categories of movement and outcomes. We compared semen quality in samples taken from 5 adult mobility (on a scale of 1-5) once per month using a subjective scoring male macaques (2 rhesus Macaca mulatta; 3 cynomolgus, M. system. Lastly, trained caregivers rate affiliative, aggressive, play, fascicularis) trained to participate in semen collection using both fear-related, and anxiety-related behaviors, as well as overall levels of methods (8 samples from each method). There was a significant wellbeing, ease of movement, activity levels, and physical health increase in sperm concentration in samples taken in the CBC each week. Every month, each chimpanzee’s mobility rating and compared to ORC (536 vs 199 million/ml, respectively; P < 0.05). progress are assessed, and new PT targets are created or increased While there was no change in motility, there was nearly a 3-fold (e.g., increase squats from 10 to 15). To date, all 9 chimpanzees increase in total ejaculatory volume (0.98 mL for CBC vs 0.35 mL for voluntarily participated in PT without hesitation. After five weeks of ORC). In addition, it took less time to train monkeys for the CBC participation, paired-samples t-tests showed that caretaker ratings of compared to ORC, and the monkeys showed fewer behavioral well-being increased significantly (P = 0.026), while ratings of ease of indices of stress. These preliminary data suggest that the closed box movement did not significantly differ (P = 0.098). These preliminary chair technique reduces stress on the animals, while enhancing the data suggest that PT using PRT is an effective refinement to captive quality and quantity of sperm samples, supporting the use of the care that allows chimpanzees to increase their mobility. This may also CBC as a refinement. be an enriching experience for the chimpanzees, as PT increases the level of human interaction and choice within the captive PS71 The Role of Positive Reinforcement Training in the Restraint environment. of Primates PS73 The Role of Positive Reinforcement Training in Veterinary JE Perlman*, M Bloomsmith Care: Improving Welfare One Grape at a Time

Yerkes National Primate Research Center, Emory University, Atlanta, E Magden* GA Department of Comparative Medicine, UT MD Anderson Cancer Positive reinforcement training (PRT) is applied across the globe with Center, Bastrop, TX nonhuman primates (NHP) in the laboratory animal science environment. PRT is used to train cooperation with husbandry, Positive reinforcement training (PRT) is used to enhance the welfare clinical, and research behaviors, including teaching NHP to tolerate of NHPs in many ways. Training the animals to participate in their some restraint behaviors such the cage squeeze back mechanism, own veterinary care not only decreases the stress often associated chair restraint, and manual restraint. Benefits of using PRT include with veterinary procedures, it also provides an enriching activity for increased cooperation with husbandry, clinical and research the animals. Sedations and anesthesia are stressful, even if we train procedures, increased welfare, decreased stress and fear, decreased the animals to present for an injection. We can minimize this stress by time to conduct procedures once training is complete, and increased using PRT to train the animals to voluntarily cooperate with their safety for animals and personnel. There are many factors that own care, often eliminating the need for sedation and anesthesia. We contribute to the successful implementation of PRT for restraint have trained NHPs to present wounds for treatment, eyes/ears/ behaviors within animal use programs. These factors include a clear throat for diagnostics, and arms/legs for blood sample collection. understanding of timelines for when the behavior is needed, support Nonhuman primates have also been trained to urinate on demand, from institutional administrators and program managers, PRT present for evaluation of blood glucose values and insulin knowledge and experience of staff members, strong communication administration, and cooperate with integrative pain-relieving among staff, effective animal training transfer and maintenance therapeutics such as acupuncture and laser therapy. Not only do plans, and understanding NHP typical and atypical behavior. these training opportunities improve the welfare of the animals, they Successful training for restraint behavior involves the use of problem are also enriching for the staff. When NHPs are trained to voluntarily solving skills and a working knowledge of how to apply PRT with provide diagnostic samples, the time required for collection decreases other techniques such as desensitization, acclimation, negative and our programs become more efficient. reinforcement, rates of reinforcement, and preference testing. PS74 The Interplay between Temperament and Operant Learning Training for restraint using PRT techniques has been demonstrated to in Primates to Cope with Medical Management positively impact animal welfare via physiological and behavioral measures and is a powerful tool in the laboratory animal science S Palmer1, R Lee1, M Niewinski1, S Oppler1, L Mutch1, J Janecek1, M field. Graham*1,2

PS72 Physical Therapy for Captive Chimpanzees Using Positive 1Surgery, University of Minnesota, Saint Paul, MN; 2Veterinary Reinforcement Training Population Medicine, University of Minnesota, Saint Paul, MN

SJ Neal Webb1,2, J Bridges1, E Thiele1, M Mulholland1,3, S Lambeth1, Training primates to cooperate with activities associated with SJ Schapiro*1,2 husbandry and medical management is paramount in improving welfare and scientific validity. The laboratory environment presents 1Comparative Medicine, UTMDACC, Bastrop, TX; 2Experimental stressors for primates that have limited experience with close contact Medicine, University of Copenhagen, Copenhagen, with humans and medical equipment. Successful training results in Denmark; 3Neuroscience, Georgia State University, Atlanta, GA enhanced coping skills and gives animals the opportunity to exercise a degree of control and choice over environment. Previous research Aging captive chimpanzees face age-related health conditions (e.g., has linked temperament in humans and primates to how they arthritis, stroke, and mobility impairment) and using behavioral respond to stressful situations. We evaluated the relationship management programs to establish personalized care routines are a between primate temperament, successful behavior acquisition, and valuable tool for the care and wellbeing of the animals. We are coping. We retrospectively reviewed data from 60 primates including implementing and evaluating the behavioral, welfare, and mobility rhesus macaques (Macaca mulatta) and cynomolgus macaques effects of a physical therapy (PT) program using positive (Macaca fascicularis) of different ages and sexes. At entry into the reinforcement training (PRT). Personalized therapy and exercise colony, primates were observed by experienced trainers who scored routines, including squats, standing, finger and toe extensions, behavioral traits which allowed categorization as either an weight shifts, and climbing were created for 9 chimpanzees with “exploratory” or “inhibited” temperament. Primates were trained mobility impairments resulting from arthritis, stroke, or injury. using mixed reinforcement training, primarily positive Chimpanzees voluntarily participate in these routines twice per week reinforcement, to perform a complex behavior (present limb to a

710 Abstracts of scientific papers 2019 AALAS National Meeting

trainer for medical manipulation in their home enclosure) in 3 parallel as expected. However, while tissue glucose decreased under phases. We found a negative association between willingness to physical restraint back to nearly baseline within 1 h, heart rates accept a treat from the hand in primates with an inhibited remained elevated until the animals were released into their home temperament at colony entry; still, inhibited animals learned at a cages. On the following d 2 to 5 tissue glucose peaks were lower each similar rate as exploratory animals. All animals completed training, consecutive day whereas the heart rate increase in the infusion chair learned the task, and demonstrated behaviors consistent with remained comparable to d 1. On d 8, when animals were replaced in productive coping evidenced by voluntary approach and the infusion chair again, after 2 d without restraint, the tissue glucose engagement with trainers. The use of a training paradigm designed peak reached comparable d 1 results again. Real-time tissue glucose to build trust and encourage animals to value rewarding over data capture with the flash glucose monitoring system is feasible in avoidance, was successful in moderating the role of temperament in macaques over at least 8 consecutive d. Glucose levels during performing a complex task as measured by total training time, total physical restraint increased in parallel with telemetric heart rate. The training sessions, and task acquisition. After controlling for other consistent increase of the heart rates indicated that at least a period of variables, we observed rhesus macaques learned faster than 5 consecutive d did not elicit any training effects reducing the stress cynomolgus macaques, and this difference was significant. level. The study design did not allow distinguishing clearly whether Considering animal attributes in relation with training methods the increase in glucose levels was clearly attributed to stress or more allows us to develop efficient behavioral management programs. likely caused by increased muscle activity. This improves the wellbeing of our primates and also the rigor of our scientific outcomes by limiting stress-confounding or introduction of enrollment bias. PS77 Light: An Extrinsic Environmental Factor that Influences Animal Health and Wellbeing PS75 Simplifying Nonhuman Primate Positive Reinforcement Training through Innovative Cage Design RT Dauchy*1, DE Blask1, LM Dupepe2, GL Dobek2, GB Voros2, AT DM Abney* Pierce2, SM Hill1

Laboratory Animal Medicine, Charles River Laboratories, Reno, NV 1Laboratory of Chron-Neuroendocrine Oncology, Tulane Univ School of Medicine, New Orleans, LA; 2Comparative Medicine, Tulane We designed and built an indoor group housing area for female University, New Orleans, LA macaques (Macaca fascicularis). It was important to design caging that would enhance welfare, facilitate animal training, and improve Light is an extrinsic environmental factor in animal facilities that efficiency by incorporating knowledge of the animals’ behavior into profoundly influences animal circadian, neuroendocrine and the cage design. There are 27 individual units, each housing up to 11 neurobehavioral regulation. Previous studies from our laboratory females that require daily vaginal swabs for menses cycle tracking. demonstrated that exposure of pigmented and non-pigmented rats to To simplify animal training, we designed a tunnel system by which light at night (LAN) of sufficient intensity, wavelength, and duration the animals enter via the back of the cage. They are trained to come suppresses nighttime pineal circadian melatonin production and to a holding cage for swabs before being released into the main unit. negatively influences metabolism and physiology. In contrast, Between June 2017 and September 2018, we received 556 females into exposure of rats to blue-enriched (465-485 nm) LED light at daytime the group housing area. All of the females were naïve to the caging (bLAD) revealed a marked positive effect on neuroendocrine and and training, but by capitalizing on their natural behavior, we neurobehavioral parameters. Here, in conjunction with our GLAS- successfully trained them how to enter the tunnel within 1–2 training supported investigations, we tested the hypothesis in mice that LAN sessions. In September 2018, data collected on 44 naïve females, disrupts, while bLAD enhances, circadian rhythms of metabolism confirmed 100% compliance with animals coming forward for and physiology associated with animal health and wellbeing. We vaginal swabs after 21 days of positive reinforcement training. examined male and female nude mice (Crl:NU(NCr)Foxn1nu; n=36 Understanding the animals’ natural behavior and incorporating it per group), commonly used in metabolism and cancer studies, and into caging design allowed for improved efficiency with regards to exposed them for 12-weeks in an AAALAC-accredited facility to animal training, husbandry needs, and research activities. either bLAD or standard broad-spectrum (300-700 nm) cool white fluorescent (CWF) light on a common lighting regimen: 12L(35.9 ± PS76 Assessing Stress Levels in Primates Using Real-time Glucose 1.3 lx [within cage];12D, lights on 0600); CWF animals were also Determinations via Cutaneous Sensors exposed to 0.2 lx (0.08 µW/cm2) CWF light during dark phase (LAN). Results showed significantly lower dietary and water intake, and J Luft*1, M Niehoff2, C Luetjens2 body growth rates in both male and female mice maintained under bLAD versus LAN(P < 0.001). Arterial plasma nighttime circadian 1VetCom, Covance Preclinical Services, Münster, Germany; 2Covance melatonin levels were over 400-fold higher in the bLAD- versus Preclincal Services, Münster, Germany LAN-exposed mice, while daily rhythms of arterial plasma total fatty acids, glucose and lactic acid levels, and pO2 and pCO2 were Does acclimatization to physical restraint procedure in an infusion significantly lower (P < 0.001) in bLAD-exposed mice. The present chair over repeated training intervals reduce the stress level in findings suggest that daytime exposure of mice to bLAD, compared primates? Continuous tissue glucose levels were evaluated and to LAN, has a profound positive impact on the circadian regulation compared with telemetric cardiovascular endpoints (heart rate and of neuroendocrine, metabolic, and physiological parameters that blood pressure) taken during repeated physical restraint in infusion influence laboratory animal health and wellbeing, and ultimately chairs. Four cynomolgus macaques where implanted with a scientific outcomes. telemetry device to obtain BP and ECG data in this IACUC-approved study. Additionally, a flash glucose monitoring system for real-time PS78 The Influence of Daytime LED Light Exposure on Circadian tissue glucose measurement was attached to the skin on the animals’ Regulatory Dynamics of Mouse Metabolism and Physiology back. Animals were dressed with a jacket to avoid interference with the device. Tissue glucose data was continuously measured and *1 1 6 4 2 remotely collected over 24 h for 8 consecutive d. Telemetric data was RT Dauchy , DE Blask , AE Hoffman , S Xiang , JP Hanifin , B 2 2 4 3 3 recorded for 2 h before, during, and after physical restraint for 6 h in Warfield , GC Brainard , M Anbalagan , LM Dupepe , GL Dobek , 4 5 4 total. To determine the effect of physical restraint, the animals were VP Belancio , EM Dauchy , SM Hill placed in an infusion chair on 5 consecutive d for 2 h and once again 1 on d 8, following 2 d rest. After animals were hand caught and Laboratory of Chron-Neuroendocrine Oncology, Tulane Univ School 2 placed in the infusion chair, glucose levels and heart rate increased in of Medicine, New Orleans, LA; Neurology, Thomas Jefferson

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University, Philadelphia, PA; 3Comparative Medicine, Tulane days for dietary/water intake, body weights, and feed conversion University, New Orleans, LA; 4Structural and Cellular Biology, efficiency. At the conclusion of the 36-day light exposure period Tulane University, New Orleans, LA; 5Department of Medicine, blood was collected over one week at 6 circadian time points (0400, Louisiana State Health Science Center, New Orleans, 0800, 1200, 1600, 2000, 2400) to obtain blood chemistries and LA; 6Epidemiology, Tulane University, New Orleans, LA complete blood counts (analysis underway). While there were no differences in light duration or within cage light intensities Light and lighting protocols, as outlined in the Guide, are important (illuminance/irradiance) between bLAD (32.16 ± 1.88 ; 78.80 ± 4.80 to both biomedical researchers and animal care personnel alike. Light µW/cm) and CWF (32.25 ± 1.88 ; 78.80 ± 1.88 µW/cm2) groups (n = entrains the master biological clock within the suprachiasmatic 36 cages/g), respectively, there were significant differences (P< 0.05) nucleus (SCN), which regulates the nocturnal pineal melatonin signal in dietary intake and body weights (P< 0.05) only in C3H male mice that temporally coordinates circadian rhythms of animal metabolism maintained in bLAD vs CWF lighting (CWF>bLAD). These data and physiology. Previous studies from our laboratory demonstrated show that daytime exposure of mice to bLAD, compared to CWF that the wavelength (color) of light impacts these responses in lighting, may influence important physiological parameters and rodents. We tested the hypothesis that daytime exposure to the most assist in our understanding regarding the impact of lighting systems. commonly used LED lighting, enriched in the blue-appearing portion (460-480 nm) of the visible spectrum (bLAD), compared to standard, PS80 In Vivo Trafficking of Immune Cells by Noninvasive broad-spectrum (300-700 nm) cool white fluorescent lighting (CWF), Molecular Fluorescence Tomography Imaging influences circadian regulation of metabolism and physiology in 3 important strains of male and female mice (C3H [melatonin- B Yang*, A Giddabasappa producing]; C57BL/6 and BALB/c [melatonin-non-producing]; n 120/group). Animals under an IACUC-approved protocol were Comparative Medicine, Pfizer, San Diego, CA maintained in an AAALAC-accredited facility for 12 wk on a common lighting regimen 12L (68.8 ± 5.2 lux; 168.6 ± 12.8 µW/ Trafficking of cells in vivo is an important phenomenon in cm2 (within cage); lights on 0600 h):12D (0 lux) on either CWF immunology and immunooncology research. In vivo tracking of cells (control) or bLAD (experimental) lighting, and were assessed at 6 after adoptive transfer or engraftment is very challenging. We have circadian time points. Compared mice housed under 12:12-h developed a method to label cells in vitro using fluorescent probes light:dark cycle in CWF light, C3H mice in bLAD evinced a 6-fold that integrate into the cell membrane without affecting cell viability higher peak nighttime plasma melatonin level (P < 0.05). C57BL/6 or function and the labelled cells can be tracked by fluorescence and BALB/c strains did not produce nighttime pineal melatonin. molecular tomography (FMT) imaging in vivo. Human T cells were Body growth rates, dietary and water intake, circadian rhythms in labelled with Cellvue815, a near-infrared (NIR) fluorescent dye that arterial blood corticosterone, insulin, leptin, glucose and lactic acid, intercalates into the cell membrane (excitation ~ 786nm; emission pO2 and pCO2, and fatty acids, and metabolic indicators (cAMP, ~814nm). The final concentration of Cellvue815 was at 50uM with 5 tissue DNA 3H-thymidine incorporation) in major organ systems million cells in 100ul PBS. Naïve NSG mice were injected with the were significantly lower (P < 0.001) in C3H mice, but not in either labelled human T cells. There were 3 mice in each group. Whole body C57BL/6 or BALB/c mice, exposed to bLAD, compared to CWF. FMT imaging was performed to evaluate the trafficking of human T Phospho-activation of major metabolic signaling pathways (mTOR, cells. In vivo (2, 24, 48, 72hr) and ex vivo FMT imaging (24, 72hr) GSK3ß, and SIRT1) was higher in skeletal muscle, and lower in liver, were taken. We showed that T-cells can be labeled by Cellvue815 for C3H mice in bLAD, compared to CWF. These data show that without affecting the viability in vitro. FMT imaging (in vivo and ex daytime exposure of C3H mice to bLAD has a marked positive effect vivo) showed that the trafficking of the labelled T cells in naïve NSG on the circadian regulation of neuroendocrine, metabolic, and mice. Quantitation showed liver and spleen as major organs for T-cell physiological parameters associated with the promotion of animal accumulation. health and wellbeing. PS81 Corynebacterium bovis: Antibiotic Susceptibility, Prophylaxis PS79 Impact of Daytime Blue-Enriched LED Light on Physiological and the Skin Microbiota Parameters of Three Common Mouse Strains Maintained on an IVC System CA Manuel*1,2, AC Fagre3, D Frank5, U Pugazhenthi4, L Johnson2, MJ Crim6, KS Henderson7, C Cheleuitte-Nieves8, DL Fong1,2, JK Leszczynski1,2, JB Daniels3, MJ Schurr9 GB Voros*1, RT Dauchy2, SM Hill2, DE Blask2, GL Dobek1 1Office of Laboratory Animal Resources, University of Colorado 1Comparative Medicine, Tulane University, New Orleans, Anschutz Medical Campus, Aurora, CO; 2Pathology, University of LA; 2Structural and Cell Biology, Tulane University, New Orleans, LA Colorado Anschutz Medical Campus, Aurora, CO; 3Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Many artificial lighting systems around the world, including CO; 4Divison of Endocrinology, Metabolism and Diabetes, University laboratory animal facilities, are transitioning from conventional cool of Colorado Anschutz Medical Campus, Aurora, CO; 5Divison of white fluorescent (CWF) lighting to light emitting diode (LED) Infectious Disease, University of Colorado Anschutz Medical lighting technology due to higher efficiency, cost effectiveness, and Campus, Aurora, CO; 6Microbiology Services, Idexx BioAnalytics, lower environmental impact. According to The Guide, recommended Columbia, MO; 7Research Animal Diagnostic Services, Charles River light illuminance for rodents is about 325 lux at 1 m (3.3 ft) above the Laboratories, Wilmington, MA; 8Tri-Institutional Training Program in floor, however, lighting technologies are not fully addressed. Laboratory Animal Medicine and Science, Memorial Sloan Kettering Previous studies from our laboratory demonstrated that the quality Cancer Center, New York, NY; 9Immunology and Microbiology, of light (wavelength) impacts metabolism and physiology in rodents. University of Colorado Anschutz Medical Campus, Aurora, CO Here we tested the hypothesis that daytime exposure to blue- enriched (460-480 nm) LED light (bLAD), the most commonly used The use of antibiotics to either prevent Corynebacterium bovis infection LED, compared to broad-spectrum CWF, influences mouse or treat infected mice has been controversial due to reports of poor metabolism and physiology. In this ongoing ACLAM-supported efficacy and fears of cultivating antibiotic resistance. However, due investigation, three common strains of age-matched male and female to the complexity of eliminating C. bovis from research vivaria, mice (C3H, BALB/c, and C57BL/6) were randomly assigned and antibiotic therapy could be a valuable tool. Accurate susceptibility maintained in an AAALAC-accredited facility on an IVC system in testing is needed to dose antibiotics appropriately. An in vitro project either CWF or bLAD lighting for a period of 36 days under an was performed to determine the minimum inhibitory concentration IACUC-approved protocol on 12L (300 lx; 123.5 μW/cm2):12D (0 lx) (MIC) of 24 antibiotics on 15 novel mouse-C. bovis isolates from light-dark cycle (lights on 0600). Animals were measured every three across the country. Then, an in vivo experiment was performed to

712 Abstracts of scientific papers 2019 AALAS National Meeting

evaluate if oral, prophylactic antibiotics can prevent C. bovis infection PS83 Longitudinal 16S rRNA Gene Sequencing of a Mouse Colony following acute exposure. An infectious dose of C. boviswas applied Associated with ‘Wild Mouse’ Gut Microbiota to 2 of 3 groups (n = 4-5/group) of singly housed, athymic nude (n = 15) and NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG, n = 14) mice. MM Hanson*, P Dube, C Horizny, J Vitale, D Gulezian, A maue Immediately following exposure one group received amoxicillin and clavulanic acid (0.375 mg/mL) in the water for 2 wk. The antibiotic Taconic Biosciences, Inc., Rensselaer, NY treated, negative and positive control groups were followed for 8 wk after antibiotic therapy by qPCR. In addition, swabs were collected The role of the microbiome in health remains a significant area of from nude mice for 16S rRNA sequence analysis to evaluate the research for many investigators with newly discovered links to impact of C. bovis and antibiotics on the dermal microbiota. Our data various disease states. Although many factors can affect the demonstrate that the MICs for all mouse-C. bovis isolates obtained do reproducibility and performance of mouse models, it is well not markedly differ from previous results published for cow-C. established that changes in the gut microbiota can have a major role bovis isolates. With the use of oral prophylactic antibiotics, neither in driving experimental variability. We hypothesized that germ-free nude nor NSG mice became infected with C. bovis following an acute C57BL/6NTac mice associated with gut microbiota collected from exposure (P < 0.05). Two weeks after infection the dermal microbiota wild mice would maintain a consistent microbial profile over time of the positive controls were significantly different from both the and generations of mice. A group of 12 mice associated at birth were negative and antibiotic treated groups and remained that way for the housed and bred for two subsequent generations within an isolator duration of the study (Q < 0.05). After 2 wk of antibiotics and 8 wk microenvironment under gnotobiotic husbandry practices. Fecal after antibiotic withdrawal, a significant difference was not observed samples were collected for 16S rRNA gene sequencing from the between the antibiotic treated group and negative control group. Our associated founder mice (n=12) and their subsequent F1 (n=40) and findings demonstrate that prophylactic antibiotics can be used to F2 (n=19) offspring at multiple timepoints. Analysis of the alpha- prevent C. bovis infections and has less of an impact on the dermal diversity of the three generations of mice harboring the ‘Wild Mouse’ microbiome than C. bovis infection. gut microbiota revealed no differences (P>0.05) in the species richness of the microbial community with respect to total number of PS82 Whole Genome, Molecular, and Biochemical Characterization operational taxonomic units (OTUs) detected. Furthermore, species of Klebsiella pneumoniae Strains Isolated from evenness and richness of the microbial community was not Immunocompromised NSG Mice significantly altered due to isolator husbandry as measured by Shannon Diversity Index. When examining specific taxonomic levels, A Mannion*1, N Fabian1, M Stair1, M Gold2, K Ribbeck2, J Dzink-Fox1, no loss of Phyla was detected during the experimental period. In S Carrasco1, E Buckley-Jordan1, JG Fox1,2 addition, the top 10 Orders accounting for greater than 97% of the total microbial community remained unchanged in terms of relative 1DCM, Massachusetts Institute of Technology, Cambridge, abundance. However, transient differences in microbial diversity MA; 2Department of Biological Engineering, Massachusetts Institute were detected among weanling mice compared to 8-week-old mice in of Technology, Cambridge, MA the F1 and F2 generations, with weanlings being enriched with Lactobacillus spp. before transitioning to a chow diet. These data Klebsiella pneumoniae (Kp) is an intestinal bacteria that can cause support that a defined gut microbiota can be stably transferred across septicemia, urinary tract infections, and pneumonia particularly in breeding generations using gnotobiotic practices. These findings may immunocompromised hosts. Some strains of Kp exhibit antibiotic improve experimental reproducibility for researchers. resistance and hypermucoviscosity, making infection challenging to effectively treat. NOD-scid gamma (NSG) mice are PS84 Novel Virus Discovery by Next Generation Sequencing immunocompromised due to defective immune cell development/ function and are used as cancer xenograft, humanized, and infectious C Wang*, P Momtsios, S Dayanis, KS Henderson disease models. In this study, NSG mice housed at a research institute experienced unexpected diarrhea, morbidity, and mortality. Kp was Research Animal Diagnostic Services, Charles River Laboratories, cultured from feces, intestines, liver, lungs, and blood at necropsy Wilmington, MA from several clinically affected mice. While Kp can cause illness in laboratory mice, mouse isolates remain poorly characterized. The Viruses are obligate intracellular parasites and can alter the objective of this study was to elucidate the pathogenic potential physiology of infected cells and communication or interaction with of Kp isolated from NSG mice by whole genome sequencing (WGS), other cell types. The existing ongoing threat of rodent viruses and the molecular, and biochemical assays. WGS was performed on 13 realization of newly discovered ones present unknown impacts on isolates cultured from the feces, cecum, liver, lung, and blood. Pan- laboratory animal research. It is imperative to have a rapid and genome phylogenetic analysis placed all isolates in a separate clade comprehensive screening of virus in research rodent colonies to most similar to human urine, blood, throat, and sputum isolates. All know the variables that may influence data obtained during isolates were assigned a multilocus sequence type of ST1165, capsule biological investigations. PCR detection of viruses is limited by K antigen of K45, and LPS O antigen of O2v2. No genomes exhibited available known virus genome diversity. Therefore genetic variation hypermucoviscosity according to the string test and were negative in undiscovered virus strains can prevent PCR detection of these for the hypermucous genes rmpA and magA. All genomes encoded viruses. Serology can only determine viral presence based on enterobactin for iron acquisition and type 1 and 3 fimbriae for available virus antigens for the detection of specific antiviral adhesion and biofilm formation. Biofilm formation was confirmed by antibody production. Virus isolation and purification is also crystal violent assay. A plasmid-encoded class 1 integron harboring challenging to establish due to the difficulty of some viruses to be antibiotic resistance genes for aminoglycosides, chloramphenicol, propagated in cell culture. Murine Chapparvovirus (MuCPV), along and trimethoprim/sulfonamide was identified in 12/13 genomes. with other novel rodent viruses, have been rapidly identified by next PCR identified this integron in 20/23 additional isolates. All genomes generation sequencing (NGS) over the last 10 y. In an attempt to also contained resistance genes for beta-lactams and fosfomycin. better understand the genetic diversity of recently reported viruses, Antibiotic resistance to beta-lactams and trimethoprim/sulfonamide we used NGS to search for additional strains. Two-4 young and adult was confirmed by MIC broth assay. In conclusion, Kp isolates from mice from a pet shop distributer were evaluated for the presence of NGS mice likely represent opportunistic pathogens. The expression newly reported viruses to assess strain variation. RNA and DNA was of plasmid-encoded multidrug resistance raises the potential of isolated from fecal samples and converted to double stranded DNA spreading antibiotic resistance to related Enterobacteriaceae colonizing by cDNA and dsDNA synthesis. DNA was further fragmented and mice housed in the same vivarium. ligated to primer linkers for sequencing. As a result, compared with GenBank viral genome sequence database, we identified novel viruses not previously reported, such as mouse alpha coronaviruses,

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Herpes virus 1, Murine Kobuvirus 2, and Murine picornavirus along sacrificing of animals. Besides, recent data reveals that the sentinel with novel viruses previously reported. These viruses shared only a health monitoring system may not efficiently and accurately 50-74% nucleotide identity to the next closest match in GenBank. demonstrate the status of ectoparasite infestations in laboratory TaqMan PCR assays were developed to determine the prevalence in animal colonies using the IVC system. In this study, we developed a research colonies. NGS is a valuable tool that broadens our multiplex PCR assay that targets the rRNA genes to simultaneously understanding of the diversity of viruses in laboratory mice. detect and differentiate these ectoparasites, including Myocoptes musculinus, Myobia musculi/ Radfordia, and the species A furmite using PS85 Genomic Characterization of Novel Mouse Kidney swab samples. This assay is very sensitive and specific with a Parvovirus strains in Laboratory and Wild Mice detection limit of 10 copies for all target agents. In 15 rodent colonies, the multiplex PCR assay has successfully detected the dual or triple Z Ge*, Y Feng, S Carrasco, S Muthupalani, D Annamalai, RM Kramer, natural infestations of fur mites in 17 of 48 rodents by fur swabs and JG Fox in 10 out of 25 rodent cages by cage swabs in 5 fur mite-positive colonies. The status of the parasite infestations in these animals was Comparative Medicine, Massachusetts Institute of Technology, confirmed with pelts examination and specific PCR assays followed Cambridge, MA by DNA sequence analyses. The rodent fur mites/actin multiplex PCR assay developed in this study could be a useful tool for Chronic nephropathy spontaneously occurs in mice. This renal monitoring rodent health in the future. pathological condition is often associated with intranuclear inclusions predominantly in immune-deficient mice, and to a lesser PS87 Improving the 3Rs in Preclinical Oncological Research wih extent, in aged immunocompetent mice including inbred C56BL/6 Multimodal Imaging and outbred Swiss Webster mice. Recently, chronic tubulointestitial nephropathy and kidney fibrosis in mice is linked to infection of an J Delgado* atypical parvovirus, termed mouse kidney parvovirus (MKPV) based on its genome sequence and organization being similar to members Fuel3D, Oxford, United Kingdom of Parvovirinae. However, determination of the complete genome sequences of new MKPV strains is essential for revealing Efficacy of treatment in preclinical oncological research is assessed by phylogenetic relationships and pathogenicity among MKPV strains. measuring tumor volume. This is measured using either medical In our study, we have identified and determined the complete imaging techniques such as US, MRI, or CT which are expensive and 4440-nucleotide genome of a new MKPV strain (MKPV MIT-WI-1) resource intensive; or callipers which are subjective and assume that isolated from Il2rg-/-Rag2-/-Wsh/Wsh mice using PCR and genome tumors are spheroidal (measuring only length and width). We have walking. The genome organization of MKPV MIT-WI-1 is identical to developed a minimally invasive, morphology independent solution previously known MKPVs. The nucleotide and deduced amino acid for subcutaneous tumor measurement using 3D imaging, sequences of the MIT-WI-1 genome is phylogenetically related to thermography, and AI. Our platform captures integrated tumor MKPV strains from laboratory mice at the Memorial Kettering Sloan volume from the 3D surface as well as 3D morphological and thermal Cancer Center (MKSCC), New York, wild mice (Mus musculus) in the features, thereby offering better performance in precision and urban areas of New York, and laboratory mice at The Centenary accuracy than callipers, but also providing extra imaging Institute (CI), Sydney, Australia. Notably, there is a dinucleotide information. With respect to tumor volume assessment, we will deletion at nucleotide 495 compared to the MKPV genome from demonstrate the extent of calliper measurement inconsistencies from MKSCC and IC. The deletion is located immediately downstream of a dataset of 2,500 tumor volume measurements from 1,600 mice (6 the coding region of NS2-P, which does not apparently affect strains), multiple operators and 20 tumor models. Then, a dataset of downstream open reading frames. Moreover, PCR and qPCR assays 2,500 scans captured this year shows 92% and 98% measurement using designed primers conserved among the known MKPV genome agreement for length and width respectively (within +/-3mm of sequences were established. Using the newly developed PCR assay, a calliper measurements) and an interoperator variance showing 93% segment of the MKPV genome, displaying 98-99% nucleotide of measurements for length and 96% for width are within 20% of sequence similarity to the corresponding regions of known MKPV repeated measurements. A further benefit is that users can be trained genomes including the MIT-WI strain, was identified in wild mice eight times faster compared to callipers. Further, we use the (Peromyscus leucopus) originating from Massachusetts Taken together, information contained in the images to estimate tumor symptoms our data indicate that MKPV is prevalent in laboratory and wild including redness, pallor, necrosis, and ulceration, as well as other mice, and provides new insights into the evolution of the MKPV biology-relevant features that can be explored as potential genomes. pharmacodynamic, toxicity, and animal welfare biomarkers. We demonstrate how these features can be useful to make preclinical PS86 Development of the Multiplex PCR Assay for Simultaneous oncological studies more statistically powerful. Our solution shows Detection of Fur Mites in Laboratory Rodents much promise for tackling refinement and reduction and will enable more confident decisions regarding animal welfare to be made in the C Wan*, C Chou early stages of drug discovery.

National Taiwan University, Taipei, Taiwan PS88 Examining the Need to Customize Animal Welfare Legislation for Animals used in Xenotransplantation Trials and Fur mites, including Myocoptes musculinus, Myobia Production musculi, Radfordia affinis, and Radfordia ensifera are the most prevalent ectoparasites in contemporary laboratory mice and rats. Recently, an M Lam*1,2 unclassified novel fur mite-like ectoparasite was identified in several rodent colonies in Taiwan. These parasite infestations are usually 1Harvard Center for Comparative Medicine, Harvard Medical subclinical; however, marked lesions, such as pruritus, alopecia, and School, Boston, MA; 2The Jeanne Marchig International Centre for ulcerative dermatitis have been reported in heavy infestation or in Animal Welfare Education, The Royal (Dick) School of Veterinary some mouse strains and may also modulate research data in animal Studies, University of Edinburgh, Edinburgh,, United Kingdom studies. Diagnosis of the fur mite infestations is thus required in routine health monitoring of laboratory animals. However, the Recent advances in overcoming both immunological and traditional diagnostic method for these ectoparasites, the subgross pathobiological barriers across species make xenotransplantation a examination of pelts, may not efficiently and accurately demonstrates potential solution to ongoing shortage of human organs. Pigs are the the fur mite infestation status, especially during light infection. best candidates so far for supplying cells, tissues, and organs for Additionally, the subgross examination of pelts will necessitate treatment of human diseases and traumatic injuries. Genetically

714 Abstracts of scientific papers 2019 AALAS National Meeting

modified pigs are being raised in the U.S. to prepare for clinical PS90 Cost Modeling: Factors to Consider when Maintaining xenotransplantation trials in humans. The Animal Welfare Act Inhouse Breeding Colonies of Mice (AWA), enforced by the United States Department of Agriculture (USDA), is the only federal law regulating the treatment of animals L Thibault* used in research in the U.S., while the Center for Biologics Evaluation and Research (CBER), under the Food and Drug Administration The Jackson Laboratory, Ellsworth, ME (FDA), has regulatory oversight of xenogenic products and xenotransplantation in humans. Animals involved in We focus on the practical considerations involved in sizing mouse xenotransplantation are not specifically described under the AWA but colonies. We demonstrate how cohort sizes can be produced more are addressed under the FDA’s “Guidance for Industry” documents effectively with a detailed understanding of productivity factors and instead. These guidance documents provide suggestions or animal health requirements. Additionally, we show how genetic drift recommendations only and do not establish legally enforceable can affect the reproducibility of experiments using mice bred in small responsibilities. They also focus primarily on public health concerns colonies. We introduce principles of cost modeling and demonstrate a rather than the welfare of source animals. Current animal protection tool that was developed to compare purchasing mice from an animal laws other than the AWA, such as antianimal cruelty laws, vary from vendor compared with breeding mice inhouse. Data shows that state to state, while animal welfare monitoring for livestock purchasing mice directly from a reputable vendor can have many production relies heavily on voluntary audits and certification benefits, including savings for an institution with regards to labor, programs. Given the unique function and needs of animals raised for time, animal space, and animal health. More importantly, use of this organ or tissue harvest for human use, producing and housing such tool can also have a positive impact on animal welfare and the 3Rs in animals will necessitate separate regulatory oversight different than certain circumstances. For example, in an experiment that requires 10 that for animals in laboratory research or food production. It is B6J female mice at 3 wk of age for a 10-wk study in an academic lab, critical for U.S. lawmakers to review, revise, and customize current we used per diem rates, cage costs, and tech costs provided by a federal and state laws to protect the welfare of this special group of major university to show how to determine the cost variance of laboratory animals. Communication and collaboration between the ordering the mice directly compared with breeding on site and the USDA and the FDA are crucial, as well as mandatory accreditation results for cost, box space, tech time, and animal welfare. This and auditing for all facilities housing animals for exampler resulted in a box savings of over 19 boxes, a decrease in xenotransplantation. As scientific discoveries lead both humans and overall costs of over $1,000, and a savings of 138 mice. Genetic drift animals toward unfamiliar domains, more rigorous and customized and the impact and number of potential mutations that occur over legislation is needed to oversee the use and welfare of animals in multiple rounds of in house breeding without genetic control is also a xenotransplantation programs. factor. Purchasing animals directly from a reputable vendor enables principal investigators to obtain animals of the correct age, strain, PS89 Behavioral and Reproductive Impacts of Environmental and sex and reduces the number of discarded animals that do not fit Enrichment and Pseudoloma neurophilia infection on Adult the criteria for the study. Zebrafish (Danio rerio) PS91 Education and Training to Fully Implement Refinement JM Estes*1, J Whitaker1, M Altemara1, MJ Crim2, C Fletcher1 Methods in Practice

1Division of Comparative Medicine, UNC-CH, Chapel Hill, K Herrmann* NC; 2IDEXX BioAnalytics, Columbia, MO Coinciding with Danio rerio’s expanding popularity in research is the Department of Environmental Health & Engineering, Johns Hopkins widespread prevalence of zebrafish microsporidiosis, a disease Bloomberg School of Public Health, Baltimore, MD caused by the parasite Pseudoloma neurophilia. Clinical signs of infection include emaciation, spinal deformities, reduced growth, Retrospective reviews of the application of refinement methods in altered behavior, gdecreased fecundity, and increased mortality. No practice revealed that the knowledge we have gained from treatment is currently available for microsporidiosis, making its refinement research is not fully applied in practice and thus, exclusion from laboratory animal facilities challenging. laboratory animals may endure unnecessary suffering. Experimental Environmental enrichment (EE) is another factor which may affect refinements, including anesthesia and peri- and postoperative behavior and reproduction in laboratory zebrafish. We investigated analgesia protocols, humane endpoints, and euthanasia methods the behavioral and reproductive effects were reviewed in over 500 basic and applied animal research that Pseudoloma neurophilia infection and EE have on anxietylike proposals. Furthermore, a literature review was conducted to behaviors and reproductive performance in adult AB strain assemble the latest best practice approaches in regards to housing zebrafish. Specific-pathogen free-P. neurophilia (SPF, n=94) zebrafish and care. In all areas of refinement, flaws in the application of and zebrafish experimentally infected with P. neurophilia (n=43) were available approaches which could help to reduce unnecessary pain, socially housed in tanks with or without EE. After 5 wk, zebrafish distress, and suffering of animals used in science were detected. One were behaviorally assessed using novel tank test (NTT) and light- potential reason that was identified in the study of experimental dark test (LDT), 2 established anxiety assays for zebrafish, and then refinement use was that researchers might not be sufficiently aware bred once weekly for 6 wk, alternating whether EE was provided in of existing refinement methods. In addition, a recent international breeding tanks each week. We hypothesized that P. survey, conducted by the European Commission’s Joint Research neurophiliainfection would increase anxietylike behavior and have a Centre found that very few courses could be identified at university negative impact on fecundity, and that the presence of EE would level that are specifically teaching the 3Rs. To avoid needless neither affect anxietylike behavior nor reproduction. Infected fish had suffering of laboratory animals, refinement methods need to be more averaged entries into the bottom of the NTT (16.6) than SPF fish sufficiently implemented in practice. Thus, an 8-wk module-based (11.3) (P = 0.003) and fish housed with EE had more averaged entries course for university students and early career scientists is currently (13.97) into the top of the test tank than fish housed without EE (8.04) being established to teach best practice approaches and to foster a (P = 0.037). No statistical differences between groups were found for culture of care for laboratory animals. Several modules cover classical LDT. SPF fish produced, on average, more eggs (17.49) and more housing and experimental refinement, others comprise refinements viable embryos at 6 d postfertilization (11.7) than infected fish (4.47 of planning, including identifying the most appropriate research and 1.06, respectively) (P = 0.0008), with EE in breeding tanks not model, proper data analysis, and comprehensive reporting. resulting in a difference in reproductive outcomes (P = 0.61). P. Consequences of poor refinement method use for the animals’ neurophilia infection and EE both affected zebrafish behavior, and welfare and for the validity of collected data are also discussed. The disease status had a significant impact on reproduction. Our findings lectures are complemented by interviews with experts in laboratory support both the exclusion of P. neurophilia from laboratory zebrafish science to explore reasons for the currently low refinement and the use of EE in housing tanks. 715 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 mplementation rates and to find possible approaches to improve the PS94 Serene Scents: Odors in Nesting Material and Plantar Sweat situation. Influence Male Mouse Social Behavior in the Home Cage

PS92 Caring for Research Animals through Final Transition: What *1 2 2 1 Can We Learn from Veterinary Hospice Practitioners? AJ Barabas , HA Soini , MV Novotny , BN Gaskill

1 2 C O’Rourke*1,2, J Welton3 Animal Science, Purdue University, West Lafayette, IN; Chemistry, Indiana University, Bloomington, IN 1Shamrock Lab Animal Consulting, Bozeman, MT; 2Peaceful Journey Pet Hospice, Bozeman, MT; 3JenWel Counseling, Bozeman, MT Severe wounding, resulting from excessive home cage aggression, is a common reason for premature euthanasia in male laboratory mice. In laboratory animal medicine, we pride ourselves on the superior Aggression can be reduced by transferring the old nest during cage care and compassion we provide for the animals we work with. At cleaning which is thought to contain aggression appeasing odors the same time, we walk a difficult balance between our attachment to from the plantar sweat glands. However, neither the deposits on used our animals and the need to emotionally detach from our situation. nesting material nor the contents of plantar sweat have been Do we honor that human-animal bond and the grief we might feel evaluated. The aims of this study were to identify and quantify when an animal must be euthanaized as part of the research study? compounds deposited in the nest site and to determine if nest and Animal hospice and in-home euthanasia is one the fastest growing sweat odor profiles correlate with social behavior. Home cage specialties in veterinary medicine. Hospice veterinarians focus on behavior was evaluated in 3 strains: SJL/JOrlCrl (high aggression), appropriate pain management, palliation of symptoms at the end of C57BL/6NCrl (moderate aggression), and A/JCr (low aggression; n life, humane euthanasia, and grief support for the owners. What can = 8/strain; 24 cages*5 mice/cage =120 mice). Aggressive and we learn from this specialty in order to better support our staff and affiliative behaviors were recorded on d 1, 2, and 7 using 1,0 the research animals with which we work? We present how the sampling. Cage hierarchy was assessed on days 5 and 6 via a tube knowledge gained in veterinary hospice can be applied to combat test. On d 7, sweat from the dominant and subordinate in each cage compassion fatigue, assist care staff through the euthanasia process, and nest samples were collected for volatile odor analysis using gas and honor the human-animal bond in a research setting. Hospice chromatography-mass spectrometry. Behavior proportions along veterinarians assist families through the end of life transition and this with nest and sweat odor were analyzed with separate principal can be applied to laboratory animal medicine. component analyses. Each significant component represents a set of odors or behaviors with high correlation. Significant components, PS93 ‘WWAG’ing Our Way towards Wellness: Implementation and from each sample type analysis, were run in general linear models to Assessment of an ASLAP Veterinary Student Canine Enrichment test if odors influence behavior. Overall, aggression had a negative relationship with nest (F =16.84; P < 0.01) and sweat (F =9.10; P < Program 1,18 1,18 0.01) odors. When there were increased nest and sweat odors, less K Nolan*, M Walker, C Hedrick, CL Freed aggression was seen in the cage. Affiliative behaviors had a positive relationship with sweat odors (F =10.91; P < 0.01). When there 1,18 ULAR, Ohio State University, Columbus, OH were increased sweat odors, more affiliative behaviors were seen in the cage. C57BL/6N cages displayed the most affiliative behaviors (F =32.36; P < 0.01; F =9.56; P < 0.01), and had high sweat odors. The Welfare and Wellness ASLAP Group (WWAG) was created to 2,18 2,18 SJLs displayed high aggression (F =117.30; P < 0.01) and had low help maximize the positive human-animal interaction opportunities 2,18 within our institution’s robust environmental enrichment program. nest odors. Previously, nest transfer has been shown to reduce This initiative provided practical hands-on experience for laboratory aggression at cage change and this data shows that odors in nesting animal career-focused students. Prior to participation, members of material and sweat correlate with low aggression and high affiliative the Ohio State University ASLAP Club completed a written behaviors. Influential odors will be candidates for future work to application to assess individual goals for the program. Following the determine their direct effect on mouse social behavior. completion of specifically tailored orientation sessions, students PS95 Increasing Public Engagement in the Evaluation of Animal completed 20-60 m enrichment sessions weekly throughout each Research: An Experiment in Transparency semester. Working in pairs, students engaged their canine

companions in leash training, forming playgroups and enhancing * canine socialization with one-on-one interactions. Volunteers MW Brunt , EH Ormandy, KE Mills, DM Weary documented session activities in a dedicated canine enrichment log to track animal-specific behaviors and preferences. A survey was Animal Welfare Program, University of British Columbia, Vancouver, distributed at the completion of the academic year to evaluate Canada individual skills, perceived benefits of the program, and acquire feedback for future improvements. Prior to program initiation, the One potential response to calls for increased openness in animal most frequently reported objectives for program participation research is to make protocols publicly accessible, but it is unclear included increased experience and comfort working with canines in a what type of input the public would provide if given this laboratory setting, positively impacting dogs through enrichment opportunity. We made five different hypothetical research scenarios sessions, and gaining more knowledge about the canine enrichment (describing projects on chronic pain research with mice, organ program. Following participation, 100% of respondents reported transplant research with pigs, smoking research with mice, their personal wellness was positively impacted by canine Parkinson’s disease research with monkeys, and cancer research with interactions and they felt their interactions positively impacted their zebrafish) available to US participants (N = 247) via an online survey. canine companions. The majority of students indicated they were Our objective was to assess participant responses to these animal more knowledgeable about the laboratory enrichment program (91%) research protocols, and identify factors that influenced acceptance or and that they had improved specific clinical skills such as low stress rejection of the proposed research. Participants were asked “Do you handling and restraint (75%) and understanding of canine behavior support the use of these animals in this research?” Response options (66%). From student feedback, we have modified our vivarium tour were yes/neutral/no, and participants were asked to provide open- and adjusted refresher training to streamline the process for ended text responses to explain the reasons for their choice. Overall, returning students. With the overwhelming success of WWAG, we 54% of the participants expressed support for the research. The have continued to grow our roster and expand our program to scenario describing smoking research with mice garnered the least include additional species. Our program exemplifies a unique support from participants (35.6%), while chronic pain research with resource for enriching our animals while fostering valuable mice was most supported (63.6%). Many participants provided open- experiences for the next generation of laboratory animal ended comments, showing that an online forum can also provide veterinarians. 716 Abstracts of scientific papers 2019 AALAS National Meeting

qualitative feedback on research protocols. Four themes were BRAD promotes peer education about animal research, offers a prevalent in participant reasoning regarding their support for the platform for those working with or supporting animals in research to proposed research: scientific merit, morality, availability of discuss their work with animals, and highlights the devoted staff alternatives, and impact on humans. These results illustrate the type who care for these animals. The event also serves as a pipeline for of public input that can emerge should research institutions provide future careers in biomedical research and laboratory animal care and opportunities for this participation as part of protocol review. We medicine. The flexible nature of BRAD allows each event to be conclude that this type public input could provide institutions a tailored to reach specific audiences and fit the resources and goals of better understanding of what types of animal research people are any organization. We discuss how BRAD has been implemented at willing to accept, and thus reduce the risk that practices fall out of an institutional level, within an entire region at multiple step with community values. organizations, and in new audiences, such as veterinary technician students and educators. PS96 Use of a Validated Smartphone-based Electrocardiogram in Mice Reveals Severe Handling-induced Bradycardia across Both PS98 Development of a Translational Canine Model of Human Sexes and Four Strains Intervertebral Disc Disease

RN Labitt*1, A Davis2, E Oxford3, S Butler4, EK Daugherity1 NN Lee*1,2, J Kramer2, A stoker2, C Pundee3, D Moore3, C Goldstein3, T Choma3, J Cook2 1Center for Animal Resources and Education, Cornell University, Ithaca, NY; 2College of Veterinary Medicine, Cornell University, 1Comparative Medicine Program, University of Missouri, Columbia, Ithaca, NY; 3Molecular Medicine, Cornell University, Ithaca, MO; 2Thompson Laboratory for Regenerative Orthopedics, NY; 4College of Veterinary Medicine, Biomedical Sciences, Cornell University of Missouri, Columbia, MO; 3Orthopaedic Surgery, University, Ithaca, NY University of Missouri, Columbia, MO

Noninvasive electrocardiogram (ECG) devices are not often used in Intervertebral disc disease (IVDD) is strongly associated with spine mice, as they require restraint or anesthesia, and expensive pain and disability without known regenerative treatments. equipment. As a result, there is little data on the effect of mouse Etiopathogenesis of IVDD is poorly understood and appears to be handling on cardiovascular parameters. We validated the use of a multifactorial. Inflammatory and degradative mechanisms are smartphone-based, single-lead ECG system in mice by performing associated with it; however, specific pathways have not been fully simultaneous ECG recordings in conscious, restrained mice and elucidated. Valid animal models that closely resemble human IVDD anesthetized mice (n=19) with surgically implanted telemetry ECG are lacking. Dogs develop spontaneous IVDD similar to humans, and systems, the gold standard for ECGs in mice. Heart rates and clinical IVDD is another significant canine health concern. This study rhythms collected by smartphone-based ECG and telemetry were was designed to test the hypothesis that pathobiology of equivalent. We observed that mice restrained in a standard intervertebral discs (IVD) from dogs and humans would show immobilizing scruff experienced an up to 80% decrease in heart rate, similarities in metabolite production and responses to cytokine equivalent to a 600 bpm decrease. We hypothesize that pressure on stimulation. With ACUC and IRB approvals, we evaluated IVDs from cervical baroreceptors produced by stretching the neck skin dorsally nonchondrodystrophic dogs (adult mongrel dogs; n=32; female n=28) during immobilizing restraint results in bradycardia, and conversely and human patients (n=85, clinical patients) to elucidate potential that restraints that do not result in pressure on the neck would not similarities and differences between canine and human intervertebral alter heart rate. Mice of both sexes and four strains (C57Bl/6, FVB/n, discs. Canine lumbar and cervical IVDs were collected aseptically DBA/2, and BALB/c, total n=38) were restrained by 3 different after euthanasia for reasons unrelated to this study. Human IVD restraint methods: a light restraint that allowed substantial head tissues were collected during surgery. Using 6 mm biopsy bunches, movement; a 3-finger scruffing technique designed to prevent IVD explants were created. Explants were separated into 2 groups, pressure on the neck by creating a transverse fold of skin; and a control and cytokine stimulated (10 ng/ml IL-1β) in supplemented standard immobilizing scruff designed to draw the forelimbs back, Dulbecco’s Modified Eagle Medium (DMEM). The explants were prevent head movement, and create a crease on the ventral neck as a cultured for 3 d and media were collected at the end of culture consequence of a longitudinal fold of skin. Three experienced mouse period. Media were tested for inflammatory and degradative handlers performed the experiments. In C57Bl/6, there was no metabolites using commercially available assays. Production levels difference in heart rate between the light restraint and 3-finger were compared for statistically significant (P < 0.05) differences. restraint method. The median heart rate using the immobilizing Significant differences in IL-6, IL-8, KC, MCP-1, MMP-1, MMP-2, and scruff was significantly decreased by 50-66% for all 3 handlers (range MMP-3 levels were noted between canine and human IVD tissues. 143-750 beats per minute). Similar bradycardia was seen in the other Human IVD tissues had higher production of inflammatory and strains and both sexes. Moreover, when the 3 restraint methods were degradative biomarkers than canine IVD tissues in both control and compared using telemetry devices, only mice that received an cytokine stimulated groups. Both canine and human IVD tissues immobilizing scruff produced intermittent delayed beats for as long showed significant increases in biomarker production in response to as 7 m after release from restraint. Due to the profound cytokine stimulation. Collectively, these data suggest that both cardiovascular effects, we recommend use of the light or 3-finger canine and human IVDs similarly respond to cytokine stimulation by scruffing technique, and avoiding or minimizing the use of the increasing production of inflammatory and degradative metabolites. standard immobilizing scruff while handling mice. Ongoing translational studies will be aimed at further elucidation of these important similarities and difference towards understanding PS97 Peer-led Outreach through Biomedical Research Awareness and addressing IVDD mechanisms in dogs and humans. Day PS99 Evaluation of Electroacupuncture for Symptom Modification AL Murray* in a Rodent Model of Human Osteoarthritis

Insourcing Solutions, Charles River Laboratories, Fredericksburg, VA AR Personett*1, M Afzali1, M Story2, R Martinez1, L Culver1, S Leavell1, A Timkovich1, J Sanford1, K Santangelo1 Accelerating strategies to increase public understanding and appreciation for essential animal research is more important now 1Microbiology, Immunology, & Pathology, Colorado State University, than ever. Biomedical Research Awareness Day (BRAD) is a day Fort Collins, CO; 2Clinical Sciences, Colorado State University, Fort devoted to honoring the role of animals in biomedical research and Collins, CO stimulating conversation about the necessary role of animals in medical advancements. Celebrated annually around the world, When faced with the frustration of chronic discomfort and restricted

717 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 mobility due to osteoarthritis, many individuals have turned to reduction in orofacial and somatic allodynia compared to TBI/no EA acupuncture as an alternative therapy. Acupuncture is a traditional group. Additionally, animals in the TBI/ EA group showed Chinese practice for pain alleviation that involves insertion of thin statistically significant improvement in physiological indices of needles into the skin and underlying tissue followed by manual spasticity compared to TBI/ no EA group. Progressively, these manipulation or electrical stimulation. However, the efficacy of studies may increase our understanding of the neurobiology of TBI- acupuncture in managing osteoarthritis is uncertain, as much of the induced spasticity and pain, and the potential mechanisms of action evidence is of questionable quality. The purpose of this study was to of experimental therapies utilizing EA. The goal of these studies is to evaluate electroacupuncture in a rodent model of osteoarthritis such enhance the opportunity for the translation of safe and effective that unbiased conclusions regarding its efficacy for symptom treatments for human TBI injury-induced spasticity and pain/ modification could be drawn. Ten 11-mo-old, male Dunkin Hartley headache conditions. guinea pigs, which characteristically have moderate osteoarthritis at this age, were randomly assigned to receive electroacupuncture (n=5) PS101 A Novel Device to Measure Static Hindlimb Weight Bearing or anesthesia only (n=5). Acupuncture points were based on the Forces in Pronograde Rodents traditional Chinese bi syndrome therapy for knee pain (BL11, BL23, BL40, BL54, GB29, GB30, GB34, GB39, LI11, and ST36). Animals were MD Williams*1,4, SL Sommer1, RC Meyers2,4, J Valdivia3, MW Nolan1, anesthetized with isoflurane, and treatments were performed 3 times BX Lascelles1,4 weekly for 3 wk. Using videotracking software, movements in an open field were recorded at baseline (prior to treatment) and then 1Clinical Sciences , North Carolina State University, College of biweekly to measure changes in activity levels as an indicator of knee Veterinary Medicine, Raleigh, NC; 2College of Veterinary Medicine, pain. Animals were harvested 2 wk after the final treatment session. North Carolina State University, Raleigh, NC; 3Honeywell, Raleigh, Serum was collected for inflammatory biomarker testing, and whole NC; 4Translational Research in Pain Program, North Carolina State knee joints were collected for histology and gene expression. Animals University, Raleigh, NC receiving electroacupuncture had a significantly greater change in total distance traveled compared to those receiving anesthesia only. Chronic musculoskeletal diseases significantly contribute to the There was a trend towards decreased serum complement component burden of persistent pain. Persistent joint pain can drastically impair 3 and tumor necrosis factor protein concentrations in the an individual’s quality of life due to alterations in limb function and electroacupuncture group compared to the anesthesia group. therefore mobility. Spontaneous pain assessment through weight Collagen type 2, fibroblast growth factor 18, and inducible nitric bearing is essential to rodent-based pain research. However, most oxide synthase gene transcripts in articular cartilage were marketed instruments assess rodents at an abnormal incline or significantly increased by electroacupuncture. There was not a during locomotion. We designed a Static Horizontal Incapacitance significant difference in total joint histology scores between groups. Meter (SHIM) for pronograde standing rodents to detect spontaneous This study provides evidence that a 3-week period of pain. We assessed the device for feasibility, repeatability, and electroacupuncture had a positive effect on symptom, but not sensitivity to quantify hindlimb bodyweight distribution. Mice and disease, modification in a rodent model of osteoarthritis. Further rats with unilateral inflammatory pain induced by Capsaicin (25 µL investigations into mechanistic pathways that may explain the of 0.1% capsaicin or 200 µL of 0.05% capsaicin, respectively) or efficacy of electroacupuncture in this animal model, as well as longer Complete Freund’s Adjuvant (CFA; 200 µL of 1 mg Mycobacterium term studies, are needed. tuberculosis) subcutaneously into the plantar surface of the left hind paw were used to measure static weight bearing. Meloxicam (1 mg/ PS100 Electro-acupuncture Prevents Spasticity and Orofacial and kg), an NSAID used for inflammatory pain, was administered to Somatic Allodynia in a Clinically Relevant Rodent Model of evaluate the ability to attenuate pain-associated weight bearing Closed Head Traumatic Brain Injury asymmetry. By evaluating acute and subchronic pain models, we validated the ability of the SHIM to detect significant reductions in P Bose2,3, J Hou2, G Mustafa2, R Nelson2, J Watts2, S Tsuda2, H limb loading on the injected hindlimb in mice and rats. Treatment Ramirez*1, F Thompson2,3 with meloxicam partially reversed CFA-induced effects. In contrast to other weight-bearing assays that measure spontaneous behavior in 1University of Florida, Gainesville, FL; 2Malcom Randal VA Medical response to a movement-related stimulus (e.g., walking), the SHIM Center, Brain Rehabilitation Research Center of Excellence (151), allows evaluation of spontaneous pain directly related to weight North Florida/South Georgia Veterans Health System, Gainesville, bearing while standing at rest. The SHIM successfully detected FL; 3Department of Physiological Sciences, University of Florida, lameness in acute and subchronic pain models and detected the Gainesville, FL analgesic effects of an NSAID. This study provides a novel tool to objectively evaluate spontaneous limb pain in rodents. Traumatic brain injury (TBI) is major health problem in both military and civilian personnel. Approximately 50% of patients diagnosed PS102 Buprenorphine administration in NZW rabbits: a with TBI end up with persistent debilitating conditions, including pharmacokinetic study of intravenous, subcutaneous and spasticity, headache, and chronic pain. Electro-acupuncture (EA) is a intramuscular administration. nonpharmacological therapeutic technique that has been shown to have a clinically relevant effect on chronic pain. The objective was to test the effectiveness of EA in the prevention of TBI-induced R Askar2, E Fredriksson1, E Manell1, M Hedeland3,4, L Olsén1, PG spasticity and painlike behaviors in a clinically relevant rodent model Hedenqvist*1 of closed head traumatic brain injury (cTBI). In this model we have shown comprehensive evidence of progressive and enduring 1Swedish University of Agriculture, Uppsala, Sweden; 2Stockholm spasticity, and orofacial and somatic allodynia. A total of 24 rats University, Stockholm, Sweden; 3Uppsala University, Uppsala, (CRL:SD) were divided in 3 groups: 1) no TBI/no EA, 2) TBI/no EA Sweden; 4National Veterinary Institute (SVA), Uppsala, Sweden and, 3) TBI/EA. EA treatment was started 30 min after cTBI as preemptive acute treatments. Treatment consisted of stimulating Buprenorphine is one of the most commonly used drugs used for acupoints ST-36, LIV-3, and LI-4 bilaterally for 30 min using postoperative pain in rabbits. Recommendations for buprenorphine continuous 10 Hz stimulation at an intensity of 2-3 mA for a total of 6 dose and dosing interval are the same for intravenous (IV), treatments over 2 wk. The velocity-dependent ankle torques, time- intramuscular (IM), and subcutaneous (SC) routes, despite lack of locked triceps surae EMGs, H-Reflex testing, and a reward-conflict comparative pharmacokinetic and sparse efficacy studies. Given that operant pain testing paradigm were applied to test spasticity and an earlier study using rabbits revealed that 0.05 mg/kg orofacial allodynia, respectively. Our data to date indicate that buprenorphine SC could not reduce pain scores after orthopedic animals in the TBI/EA group exhibited a statistically significant surgery, an experiment was conducted to assess the following

718 Abstracts of scientific papers 2019 AALAS National Meeting

hypotheses: 1) the AUC would be lower for SC than for IV and IM Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, injections, 2) that the bioavailability (F) would be lower after SC than PA; 2Neurosurgery, University of Pennsylvania, Philadelphia, PA IM injection, 3) that Cmax would be lower after SC than IM injection, and 4) that a higher SC dose could be used to compensate for a lower The pathogenesis of neurodegenerative diseases, such as F. Five female and 5 male NZW rabbits (3.1± 0.3 kg) were spinocerebellar ataxia, involves genetic defects in specific cell administered buprenorphine SC (0.05 and 0.1 mg/kg), IV and IM populations within the brain. A major challenge for gene therapy (0.05 mg/kg) in a randomized order with 2 weeks wash-out between research aimed at correcting these defects is developing methods administrations. Serum was analyzed with HPLC-MS before, and up capable of delivering therapeutic genetic material to cells most to 480 min after injection. Data was analyzed with 2-way ANOVA affected by the disease process being studied. Here, we outline a with administration route and sex as treatment factors, animal and novel method of delivering adeno-associated viral vectors (AAVs) to time as blocking factors, and body weight (BW) as a covariate. Purkinje cells in the cerebellum of rhesus macaques (Macaca mulatta) Administration route had an overall effect on all parameters, an using a frameless stereotactic device under MRI guidance. Isoflurane- affect that varied between sexes. AUC was significantly lower for SC anesthetized animals are placed in the bore of a MRI machine. 0.05 than: (i) IV in females (423 ± 148 and 725 ± 211 µg/L*min, P = Percutaneous pins stabilize the skull and a specialized surgical drape 0.003) but not males (423 ± 149 and 530 ± 163 µg/L*min, P = 0.086), that maintains sterility of the prepared surgical site as the animal (ii) IM in females (773 ± 185 µg/L*min, P = 0.004) but not males (634 moves in and out of the bore during the procedure is placed. An ± 141 µg/L*min, P = 0.026), (iii) SC 0.1 in females (633 ± 211, P = MRI-compatible disposable trajectory device, which houses a guide 0.015) and in males (958 ± 387 , P < 0.001). The bioavailability was tube containing a liquid contrast agent, is mounted to the skull with significantly lower for SC 0.05 than: (i) IM in both females and males percutaneous bone screws. To avoid disturbing vasculature in the (50 ± 20 and 95 ± 21 %) and (ii) SC 0.1 in males (71 ± 23 %) but not brain, liquid contrast is delivered intravenously prior to trajectory females (36 ± 14 %). Cmax was lower for SC 0.05 than: (i) IM in both planning. Using MRI imaging combined with a specialized trajectory sexes (2 ± 1 and 10 ± 4 ng/mL, p<0.001) and (ii) SC 0.1 in males (7 ± 3 mapping software program, individual nuclei within the brain are ng/mL, p<0.001) but not females (3 ± 1 ng/mL, p=0.015). In targeted for direct AAV delivery. The trajectory is plotted and the conclusion, a dose for SC administration of buprenorphine could not device is positioned in line with the target. A small burr hole is be recommended due to the low bioavailability which is not simply created in the skull, followed by introduction of a cannula to a depth compensated for by increasing the dose. determined by the mapping software. The trajectory of this cannula is confirmed with MRI imaging prior to the commencement of a slow PS103 Comparative Risk of Human Injury/Exposure while infusion of a phosphate-buffered saline solution containing the Collecting Blood from Sedated and Unsedated Nonhuman therapeutic agent and a contrast agent. Postinfusion imaging Primates confirms that the solution adequately covered the target. The cannula is retracted after a dwell period adequate to minimize reflux up the C Hotchkiss*, M Young cannula path, and the skin is closed with suture. Prior to recovery, all hardware is removed. This technique may be used to deliver Washington National Primate Research Center, University of therapeutic agents to multiple targets within the same animal, and it Washington, Seattle, WA may be used in other large animal model species, such as pigs and dogs. Collection of blood samples for research or clinical evaluation is one of the most common procedures performed in nonhuman primates. PS105 Reducing Variability in Mouse Studies through Continuous There are several possible methods to obtain samples. In the early Data Collection days of primate research, manual/physical restraint was used, which was stressful for the animal and risky for the human. As the field L Schaevitz*, M Lim, E Defensor developed, chemical immobilization with ketamine or other anesthetics became the most commonly used method. More recently, Vium, San Mateo, CA the use of training using positive reinforcement has allowed collection of blood samples from unsedated primates that are Inconsistencies in the reported behavioral phenotypes of mouse unrestrained or minimally restrained. Elimination of anesthesia models present challenges for scientific interpretation. Behavioral reduces risks to the animal. We wanted to know whether there was a traits are challenging to measure consistently due to the influence of difference in risks to humans between the 2 methods of blood interacting genetic and environmental factors. As a result, the collection. We evaluated injury and near-miss reports in conjunction reported behavioral phenotype of a model can vary widely between with blood collection data from 2011 to 2019 at a primate center that laboratories and likely stems from conditions which are difficult to houses macaques (M. nemestrina, M. mulatta, and M. fasicularis) and standardize between labs. In an effort to identify approaches to squirrel monkeys (S. sciureus). Injuries associated with sedated blood improve the consistency of findings, we hypothesized that collection included those occurring during the sedation procedure continuous collection of data in the home cage would optimize and recovery, as well as those directly associated with blood phenotyping by reducing the impact of experimental confounds. The collection. Injuries associated with unsedated blood collection hypothesis was tested in the cuprizone mouse model which is used included those which occurred both during animal training and to study de/remyelination associated with motor deficits. Results during blood collection. Overall, there were 16 injury exposures and from standard behavioral assays are highly variable with reports of 3 near misses associated with 62,522 blood collection procedures and open field activity that is increased, decreased, or normal in 0.022% of sedated blood collections were associated with exposure cuprizone-treated mice. In this study, female C57Bl/6J mice received incidents and 0.088% of unsedated blood collections were associated either control chow (n=6) or 0.2% cuprizone chow (n=6) for 6 wk. with exposure incidents. Our data indicate a very low risk of Throughout the study, mice were single-housed on racks with cloud- exposure associated with blood collection. The risk was higher for connected video cameras. In the cloud, computer algorithms unsedated animals (P = 0.04 by chi square analysis), but the low processed the video to create a motion biomarker. Analysis of motion number of incidents makes statistical interpretation questionable. revealed variable responses depending on the time of day and occurrence of recent procedures (cage change). Nighttime motion PS104 A Novel Method of MRI-guided Gene Therapy Agent was significantly decreased in cuprizone vs. control mice on nights Delivery to Nonhuman Primate Brain Using a Frameless 5-14, 20-28, and 35-41. Daytime motion was not different with the Stereotactic Device exception of significantly elevated motion in cuprizone mice following cage change. Continuous monitoring showed that different A Muehlmatt*1, TH Lucas2, GR Smith1, MK Keiser1, BL Davidson1 conditions throughout the study, such as time of day or procedure, interact with the cuprizone treatment to produce different behavioral 1The Raymond G. Perelman Center for Cellular and Molecular outcomes. These results reinforce our understanding that traditional

719 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 behavioral tests, utilizing a limited observation window, are B6-db) and a C57BLKS/J strain with sustained hyperglycemia (BKS- influenced by environmental and procedural factors. In contrast, db). On a standard chow diet, serum cholesterol measured on continuous data collection allows for a more comprehensive chemistry analyzer was higher at 8 wk in Apoe-null males (531 vs 470 characterization of an animal model since transient disturbances are mg/dL; P = 0.0036; n=20 per sex) but did not differ significantly by detected as anomalies within a larger data set. The results highlight sex in Ldlr-null (P = 0.9673; n=20 per sex); male and female the utility of continuous collection of undisturbed behavior for better cholesterol was higher than sex-matched controls (P < 0.0001 for characterization of motor abnormalities in disease models. both). LDL did not differ by sex in Apoe-null mice while LDL was higher in Ldlr-null females (females: 88 vs 65 mg/dL; P = 0.0006). PS106 Exploring Cardiovascular and Metabolic Stress Responses to HDL and ApoA1 were elevated in Apoe- and Ldlr-null and C57BL/6J Carbon Dioxide Euthanasia in Rats males (P < 0.001 for each sex-wise comparison). As assessed by histopathology, atherosclerotic plaque formation varied by strain but not sex. In the diabetic strains, homozygous males and females KE Nichols*1, JM Doyle2, KL Holliday-White1, KM Swearingen1, HM gained weight similarly despite sexual dimorphism in control Bogie1, M Swaab Fine3, SR Tiesma4 weights. In homozygous mice at 8 and 16 wk of age, the only significant weight differences between males and females (n=40 of 1Surgical Services, Data Sciences International, St. Paul, each) were at 8 wk in B6-db (males were 1.7 g more; P = 0.0037) and MN; 2Services, Data Sciences International, St. Paul, MN; 3Boston 16 wk in B6-ob (females were 2.5 g more; P = 0.0017). BKS-db/ Scientific, St. Paul, MN; 4Marketing, Data Sciences International, St. db males had higher blood glucose than females (415 vs 357 mg/ Paul, MN dL; P < 0.0001) at 8 wk but no other differences were found at 8 and 16 wk. HbA1c was higher in males for all strains at 16 wk (P < 0.05 Euthanasia is a necessary component in research and must be for all comparisons). As assessed by DEXA imaging, male and female conducted as humanely as possible. Carefully regulated homozygotes had similar body fat despite differences in control CO2 exposure in conscious rats is currently recommended but genotypes. BKS-db/db insulin was higher relative to controls at 8 wk, published data are divided on whether this method is more but db/db levels did not differ by sex (n=10 per sex). Leptin was distressing than others, such as adjunct anesthetic use prior to elevated in db/db mice over controls (P < 0.0001 for all comparisons). CO2 exposure. Thus, we evaluated cardiovascular and metabolic Leptin did not differ by sex in db/db mice at 8 wk, but was higher in responses of direct CO2 versus isoflurane exposure prior to CO2 to 16-wk females (P = 0.0039). All data are in Mouse Phenome Database assess distress in rats during euthanasia. We hypothesized that mean as references for using female mice in diabetes and cardiovascular arterial pressure (MAP), heart rate (HR), and plasma glucose (GLU) research. would initially increase as a stress response to either gas and that these increases would be more marked with CO2 alone. Male PS108 Subcutaneous Mass in a Naked Mole Rat (Heterocephalus Sprague Dawley rats were implanted with telemetry devices to glaber) obtain MAP, ECG, and GLU. Animals recovered for two weeks and * were exposed to either 5% isoflurane (Iso+CO2, n=6) or 100% KA Coda , C Adams CO2 (CO2exp, n=7; 30% chamber volume replacement) in their home cages to induce loss of consciousness (LOC). Once LOC was achieved University of Illinois, Chicago, IL with isoflurane, Iso+CO2 rats were transferred to a charged CO2 chamber for euthanasia. CO2exp rats continued to receive CO2 in A 9-mo-old experimentally naive female naked mole rat (NMR) their home cage. MAP, ECG, and GLU were continuously acquired (Heterocephalus glaber) presented with an approximately 2.5 x 1.0 x 0.5 prior to gas exposure through death. MAP increased in both the cm soft to firm, dark purple to black subcutaneous mass on the right CO2exp and Iso+CO2 groups when exposed to CO2 or isoflurane, ventrolateral abdomen. Otherwise the NMR displayed no adverse respectively. MAP in the Iso+CO2 group decreased during LOC and clinical signs, was in good body condition, and displayed normal further dropped when exposed to CO2 while MAP in the CO2exp behavior. The NMR was from a closed colony that have been bred group gradually increased until death. HR was elevated during inhouse for approximately 20 y and have routinely tested negative initial isoflurane exposure and returned to baseline before for common rodent pathogens. The differential diagnosis list CO2 exposure. When exposed to CO2, HR dropped dramatically in included neoplasia, hematoma, or abscess. Naked mole rats are a both groups. GLU remained stable throughout LOC with isoflurane species known for their longevity and resistance to neoplasia. There and decreased slightly and transiently when placed in CO2. GLU are only very rare reports of neoplasia in the literature. Similarly, the decreased in the first minute after CO2 exposure in the CO2exp group young age of the animal also reduced the likelihood of neoplastic and levels were stable until death. Time to death was shorter in disease. Fighting or traumatic injuries are one of the more common CO2exp. Isoflurane exposure caused increased MAP and HR, health issues reported in this species, which could lead to either a whereas direct exposure to CO2 caused an increase in MAP but not in hematoma or abscess. However, no external wounds were seen on HR. A mild decrease in GLU was detected in CO2exp. These data physical examination. The NMR was euthanized by anesthetic suggest that LOC with exposure to either gas may be distressful, but (ketamine/xylazine) overdose and a necropsy was performed. Gross euthanasia via CO2 without preanesthetic appears to occur quicker necropsy revealed that the large, dark subcutaneous mass was the and elicit less of a physiologic stress response. cecum, which had herniated through a defect in the abdominal wall. The abdominal hernia was likely a result of a congenital defect or PS107 Comparing Male and Female Phenotypes in Common trauma, though no traumatic events were noted by animal care or Mouse Models of Cardiovascular and Metabolic Disease investigative staff.

J Serrano1, K Leighton2, R French2, A Schile*1 PS109 Generalized Neurologic Disease in an Athymic Nude Mouse

1The Jackson Laboratory, Sacramento, CA; 2The Jackson Laboratory, W Hanson*1, U Blas-Machado2, V Lee1 Bar Harbor, ME 1Division of Animal Resources, Emory University, Atlanta, National Institutes of Health policy states that animal studies should GA; 2Department of Pathology, University of Georgia, Athens, GA account for sex as a biological variable by including males and females when possible. We studied key phenotypes in mouse disease A 15-month-old, singly housed, female athymic nude mouse was models with a large usage bias toward males: cardiovascular disease examined for acute hindlimb lameness. The animal was strains (C57BL/6J mice with Apoe or Ldlr mutations), and diabetes experimentally naïve and last observed to be normal 1 wk prior. On models with inactive leptin (Lepob) or its receptor (Leprdb): two physical exam, the mouse was bright, alert, responsive, hunched, and C57BL/6J mutant strains with transient hyperglycemia (B6-ob and slightly dehydrated and thin. The mild dehydration resolved with

720 Abstracts of scientific papers 2019 AALAS National Meeting

moist chow provided on the cage floor, and eating, drinking, as leiomyosarcoma. Medical oncotherapy was not pursued due to urination, and defecation were all observed to be normal. On practical limitations. While mass regrowth is expected due to the neurologic exam, there was moderate kyphosis and decreased weight histopathologic diagnosis, no regrowth was found 4 mo after surgery. bearing in all limbs. The mouse generally remained in a sitting Leiomyosarcomas of the skin and subcutis are rarely reported in position with hindlimbs splayed, and nonambulatory tetraparesis avian species. was observed. Cranial nerves were within normal limits, and paw placement was intact. Withdrawal reflexes were decreased, and PS111 Unusual Cause of Hind Limb Paresis in C57BL6 Mice Used hindlimb splay reflexes were absent. Superficial pain was positive in for a Behavioral Study all limbs. On orthopedic exam, all limbs had decreased range of motion and mildly decreased muscle mass/tone. No overt pain or SA Kurnick*, RM Kramer, S Muthupalani soft tissue swelling was noted, but the hindlimbs appeared mildly erythemic. At the stifles, 2-3 small, white, firm, round nodules were Division of Comparative Medicine, Massachusetts Institute of present. Clinical signs and examination suggested the presence of Technology, Cambridge, MA both a lower motor neuron disease as well as an orthopedic condition. Differential diagnoses for the neurologic disease included An experienced investigator noted that an entire cohort (n=6) of neuritis, polyradiculoneuritis, acute inflammatory demyelinating young male C57BL6 male mice exhibited varying degrees of hind polyneuropathy, peripheral nerve sheath tumor, paraneoplastic limb paresis following short habituation sessions in a restraint device polyneuritis, and hypermagnesemia. Differential diagnoses for the for head-fixed, awake, behavioral experiments. The clinical history orthopedic disease included osteoarthrosis, osteomyelitis, myositis, revealed that 2 wk prior to the behavior habituation, animals osteoporosis, muscle atrophy, and osteosarcoma. The animal was received a craniotomy and head-plate implant surgery. All animals euthanized and submitted for histopathology which revealed acute, recovered uneventfully and were clinically normal postsurgery until multifocal, moderate polyradiculoneuritis of the nerve roots and habituation in the restraint device. This investigator had performed ganglia. It also revealed chronic, moderate osteoarthrosis of the both the surgery and behavior habituation procedures in mice femorotibial joints with cruciate ligament and menisci involvement. previously with no issues. Several animals demonstrated only mild While nearly all strains of mice develop some form of osteoarthrosis improvement after several days and physical examinations revealed with age, acute polyradiculoneuritis is an unusual condition in normal mentation with no clinical abnormalities other than hind limb laboratory mice. This disease shares many clinical similarities with ataxia. Two morbid animals were euthanized for detailed necropsy. Guillain-Barré syndrome in humans, which has been associated Grossly, the skin of the dorsum on both mice was slightly depressed with Campylobacter jejuni infection. Clinicians should consider the and irregular over an area of 2 cm x 0.5 cm along the spine. On potential bacterial etiologies of these conditions in mice, such as dissection, the underlying subcutaneous tissue and musculature was from C. jejuni. necrotic and firm.Histologically, in both cases there was regionally extensive acute to subacute necrosis, degeneration, edema, PS110 Cervical Cutaneous Mass in a White Carneau Pigeon mineralization, and atrophy with associated acute neutrophilic (Columba livia) inflammation of the skin, subcutis, and underlying musculature. These findings extended to the distal cervical-thoracic vertebral *1 2 1 1 1 N Fabian , M Esmail , JL Haupt , S Muthupalani , S Carrasco bones with degenerative myelopathy of the underlying spinal cord consistent with thermal/caustic injury. On follow up queries, the 1 Division of Comparative Medicine, Massachusetts Institute of investigator revealed that he had used a halogen lamp to illuminate 2 Technology (MIT), Cambridge, MA; Tufts CMS (Comparative the head-plate for 5-10 m in close proximity to the animals at the Medicine Services), Tufts University, Boston, MA onset of each habituation session. On the basis of the experimental history as well as the clinical and pathological findings, the cause of A 14-y-old male research White Carneau pigeon used in psychology hind limb paresis was determined to be halogen lamp-induced studies was assessed for a soft tissue mass on the right ventral neck severe deep thermal injury with myelopathy. This case demonstrates near the crop. Two months prior, a mass suspected to be a feather the importance of creating awareness among personnel manipulating follicle cyst 1cm in diameter was noted at the same location. An open laboratory animals regarding types of light sources and their wound over the mass and blood on the bird’s beak indicated self- potential to cause thermal injury. trauma. The next day, the mass was cavitated, suggesting that a portion of it was pecked and removed by the pigeon. Medical care PS112 Sudden Death after Laceration Repair in a Pair- included oral trimethoprim sulfamethoxazole, meloxicam, and Housed C1Q Transgenic Rabbit bandage application. Two months later, the mass was 3-4x larger (4.0 x 3.5 x 2.5cm), soft to firm, freely moveable, and extended into the C Hunter*1, PA Lester1, J Xu2, J Zhang2, J Song2, D Yang2, I Bergin1 muscularis. Physical examination did not reveal other masses or abnormalities. CBC and biochemistry were unremarkable. Metastases 1Unit for Laboratory Animal Medicine, University of Michigan, Ann were not seen on whole body radiographs, which were normal aside Arbor, MI; 2Internal Medicine-Cardiology, University of Michigan, from the mass. On ultrasound with color flow Doppler imaging, the Ann Arbor, MI mass was highly vascular with a multilobular nodular pattern. The mass was excised under general anesthesia, but complete surgical A 13-mo-old, pair-housed, intact male rabbit was found dead in its margins were difficult to obtain due to close adherence to the cage 3 d after treatment of minor skin lacerations suspected to be esophageal serosal surface. Recovery was uneventful and the bird fight wounds. This rabbit was a C1Q heterozygous knockout. C1Q is resumed experimental operant chamber testing 11 d after surgery. a classical complement component and its insufficiency is associated Histologically, the dermis and subcutis were expanded by a well- with autoimmune disease including systemic lupus erythematosus demarcated, nonencapsulated, densely cellular mass comprised of (SLE). Skin ulcerations on the ear and dorsum had been clipped and sheets and haphazardly arranged short-interwoven bundles of cleaned under inhalant anesthesia, and the animal had been neoplastic spindle to elongate cells supported by a thin fibrovascular separated from its male cagemate and treated with subcutaneous stroma. Neoplastic cells formed variably sized nodules surrounding meloxicam and topical silver sulfadiazine. Differential diagnoses for dilated veins, arteries, and cystic cavitations containing necrotic sudden death included adverse anesthetic/analgesic reactions, debris that extended to deep and lateral surgical margins. Neoplastic hemorrhage or infection of wounds, stress-related intestinal ileus, or cells exhibited moderate to marked atypia and 91 mitotic figures per glomerulonephritis due to C1Q insufficiency. Necropsy findings 10 HPFs. On immunohistochemistry, neoplastic cells were positive included acute necrotizing pancreatitis and focal duodenal for desmin and α-smooth muscle actin and negative for perforation with fibrinous, nonsuppurative peritonitis, adrenal gland pancytokeratin and S100. Based on gross, histological, and infarction, and extensive submucosal gastric amyloidosis. Abdominal immunohistochemical features, the cutaneous mass was diagnosed fluid cytology showed a predominately lymphocytic infiltrate. Taken

721 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 as a whole, the precipitating cause of death appeared to be acute, tumor growth and elected to continue monitoring this mouse. Two w severe pancreatitis and duodenal perforation, but the antecedent after initial presentation, the lab decided to euthanize the mouse due cause was unclear. Both gastric amyloidosis and pancreatitis are to weight loss of 2 grams. The list of differential diagnoses included unusual in rabbits. Cutaneous, renal, or other manifestations of SLE hemolytic disorders, hepatitis, cholangitis, cholelithiasis, were not present. C1Q mutations have been associated with other cholecystitis, and neoplasia. On necropsy, the gallbladder was autoimmune disorders including scleroderma and urticarial severely distended at 7mm in diameter; the gallbladder contained cutaneous vasculitis. Unfortunately, serum samples and skin lesions mildly viscous, dark green/black bile with no evidence of choleliths. were not available in this animal. While spontaneous pancreatitis Clinical chemistry revealed elevation of leakage and induced liver cannot be ruled out, its infrequency in rabbits and the concurrent enzymes, hypercholesterolemia, and hyperbilirubinemia. On presence of amyloidosis, adrenal gland infarction, and skin lesions histopathology, we found evidence of metastatic carcinoma in the suggest that a C1Q-related autoimmune component may be present. gallbladder, lung, pancreas, ovaries, uterus, mesenteric, and peri- This case illustrates that sudden death in transgenic animals can act pancreatic lymph nodes. The liver also had evidence of as a sentinel event that may guide investigation of potential cholangiohepatitis and coagulative necrosis. Based on the lack of phenotypes. Biopsy of unusual skin lesions, complete tissue neoplastic foci on serosal surfaces, inadvertent intraperitoneal collection from animals found dead, and assessment of serum injection was unlikely. This case represents an atypical presentation autoimmune antibodies will be useful in following this colony going of subcutaneous tumor implantation. forward. PS115 Abdominal Mass in a Female Common Marmoset (Callithrix PS113 Complications following Caesarian Section in a Common jacchus) Marmoset (Callithrix jacchus) SJ Mangosing*1, C Echeverri1,2, A Piersigilli1,3, C Pressl2, LL Diaz1,2 J Brockhurst*, JA Herrod, CF Brayton, JM Izzi 1Tri-Institutional Training Program in Laboratory Animal Medicine Molecular and Comparative Pathobiology, Johns Hopkins University and Science, New York, NY; 2The Rockefeller University, New York, School of Medicine, Baltimore, MD NY; 3Laboratory of Comparative Pathology, Memorial Sloan Kettering, Weill Cornell Medicine, & The Rockefeller University, New A 2-y-o, pair-housed, primigravid common marmoset (Callithrix York, NY jacchus) presented for dystocia. Two stillborn fetuses were delivered An adult, experimentally naïve, multiparous female common following medical intervention. Radiographs and abdominal marmoset (Callithrix jacchus) presented with a large, approximately 3 ultrasound revealed 2 remaining dead fetuses, which were removed x 4 x 2 cm, firm, immobile mass in the right cranial abdomen during through caesarian section. On postoperative d 1, the marmoset quarantine. Diagnostic test results and the clinical evaluation were developed swelling, erythema, and lameness of the right leg below otherwise unremarkable. The animal was cohoused with a male. the stifle. Differential diagnoses at this time included ischemia- Ultrasound revealed the presence of a hyperechoic mass with solid reperfusion injury secondary to catheter placement or the leg-tie and fluid-filled compartments in the right cranial quadrant of the during surgery, venous thrombosis, inferior vena cava syndrome, abdomen. MicroCT revealed an intraabdominal hyperdense fetiform cellulitis, or trauma. Blood work revealed a marked structure in the region of the palpated mass. Differential diagnoses thrombocytopenia (PLT 22K/µL), elevated BUN, and included teratoma, ectopic pregnancy, or fetus in fetu. The animal was hypoproteinemia. Supportive care, analgesia, and systemic euthanized after an acute terminal procedure and submitted for antibiotics were initiated. Right leg swelling and skin necrosis complete necropsy. On gross examination, a large, oval heterogenous progressed with subsequent loss of motor function, deep pain mass was found adhered to the margins of the left liver and sensation, and appreciable tarsal pulse. The distal tail and left foot abdominal wall. Macroscopically, the mass’s consistency ranged from also developed swelling and erythema approximately 48 h after soft to hard depending on location. On cut section, it appeared to surgery. Due to rapid deterioration and tissue compromise, a mid- contain different tissue types with a variable level of organization. femoral amputation of the right leg was performed. The marmoset Microscopically, the mass had a thick fibrous capsule and contained recovered well following the second surgery, but the distal tail and bony structures surrounding 2 skull-like structures, as well as left foot became necrotic over the next several days. Euthanasia was segments of axial and appendicular skeleton, soft tissues and organs. elected due to poor prognosis. Necropsy and histopathology MicroCT, anatomic, and histologic findings, differential diagnoses, confirmed ischemic necrosis of the left foot and distal tail, with and prevalence in nonhuman primates will be discussed. thrombi near the junction of necrotic and healthy tissue in the tail. Few foci of necrosis and scarring consistent with infarction were PS116 Dyspnea in a Long-Evans Rat (Rattus norvegicus) present in both kidneys. The cause of morbidity in this patient was likely multifactorial. Hypercoagulability associated with pregnancy A Skorupski*, I Bergin, R Dysko and compression of the vena cava while supine during C-section surgery, both of which are common in humans, may have Unit for Lab Animal Medicine, University of Michigan, Ann Arbor, contributed to decreased peripheral perfusion and subsequent injury MI to the extremities. A 3-mo-old male Long-Evans rat (Rattus norvegicus) presented PS114 Icteric Mouse in a Tumor Xenograft Study acutely with open-mouthed breathing. The only experimental manipulation was an intravitreal injection of an adenovirus vector CK Gow*1, AL Johnson2, KE Saunders1 one month prior. On physical examination, the rat was bright, active, and responsive, and in addition to the dyspnea was sneezing with 1Department of Comparative Medicine, Oregon Health & Science production of white foamy nasal discharge. Differential diagnoses University, Portland, OR; 2Department of Comparative Medicine, included upper respiratory infection, pneumonia, or congestive heart Oregon National Primate Research Center, Beaverton, OR failure. Euthanasia was elected because of poor prognosis. Gross necropsy findings were pulmonary hyperinflation, esophageal and A 4-mo-old, 20.3g, female athymic nude mouse (Crl:NU(NCr)- intestinal dilation suggestive of aerophagia, and a markedly Foxn1nu) was examined for concerns of yellow discoloration to the thickened left cardiac ventricle. Histologically, there was extensive skin (icterus). Two mo prior, the lab injected 4-6 x 106 MDA-MB cells cardiac myocyte hypertrophy but without significant degeneration, subcutaneously in the right flank of the mouse. On initial necrosis, inflammation, or fibrosis. Neither pulmonary edema nor presentation, the physical exam was unremarkable except for the hepatic congestion were present. Thus, although the animal had yellow tint to the skin; the animal›s abdomen palpated soft and severe hypertrophic cardiomyopathy (HCM), the clinical nonpainful with no evidence of masses or fluid. The lab did not note presentation did not appear to be due to acute cardiac

722 Abstracts of scientific papers 2019 AALAS National Meeting

decompensation. There was, however, severe rhinitis and necrotizing changing behaviors and practices in those working with research laryngotracheitis with mixed gram-positive and gram-negative rodents. This study focused on the need for understanding principles bacteria. Warthin-Starry stains for CAR bacillus were negative. of human behavior change to improve animal welfare. The specific Tracheal culture yielded a mixed population of alpha objective was to assess the current attitudes of laboratory animal hemolytic Streptococcus, Escherichia coli, Enterococcus professionals, including veterinarians, veterinary technicians, and faecalis, coagulase-negative Staphylococcus spp., and Proteus spp. Based animal care personnel towards research animal welfare to determine on histologic severity and the presence of mixed commensals, the major animal welfare concerns for rodents. Study participants were acute presentation appeared due to the severe rhinitis and recruited from amongst the memberships of CALAS and CALAM. laryngotracheitis, but a primary pathogen was not identified. Methods consisted of 1:1 structured 30-60-m interviews with 17 Although the inciting cause of infection is not known, increased veterinarians and 18 veterinary technicians/animal caregivers from respiratory effort due to preexisting HCM may have played a role a range of research facility types from across Canada. Volunteers due to inhaled nasal irritants (ammonia or bedding material). HCM were asked between 13-17 questions, depending on their responses, has been reported in certain rat strains, but the heritability and and they were additionally asked to complete a 5-question written prevalence in rats overall is unknown. This case presentation was demographic survey after the oral interview. Interviews were striking since the clinical signs could have been attributable to either recorded and transcribed verbatim and transcripts were subsequently respiratory or cardiac conditions and the case turned out to have coded using a thematic analysis. Our results identified no discernible elements of both. differences in response comments between gender, age, geographic distribution or institutional type, although responses did differ PS117 Cutaneous, Oral, and Abdominal Masses in a Cohort of in weighting between veterinarians and veterinary technicians/ Immunomodulated Syrian Hamsters (Mesocricetus auratus) animal caregivers. Both groups consistently identified current rodent housing standards and pain management as their biggest welfare IA Galex*1, AD Otero Segui1, JM Estes1, RJ Pickles2, VL Godfrey1, VK concerns. Historical standards and speciesism were identified as the Baxter1 primary reasons for current welfare issues, but most interviewees remain optimistic that it is possible to move beyond these limitations. 1Division of Comparative Medicine, University of North Carolina at We hope to use these findings to develop a general action plan that Chapel Hill, Chapel Hill, NC; 2Department of Microbiology and may be used to address research rodent welfare issues. Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC PS119 Cognitive Dissonance in Laboratory Animal Medicine and Implications for Animal Welfare A cohort of 20 adult male and female STAT2 knockout Syrian hamsters (Mesocricetus auratus) was imported from another academic RM Engel*1, CC Silver2, C Veeder3, RE Banks4 institution. Approximately 8 wk after arrival, and prior to the start of an experimental infectious disease study, 6 hamsters presented with 1Division of Veterinary Resources, National Institutes of Health, multiple swellings on the face and/or legs or with poor body Bethesda, MD; 2SoBran Biosciences, Bethesda, MD; 3Fox Chase condition and an enlarged abdomen with a firm, palpable mass. The Cancer Center, Philadelphia, PA; 4University of Oklahoma Health animals were euthanized and necropsies were performed. Gross Sciences Center, Oklahoma City, OK necropsy findings varied among the hamsters and included exophytic growths on oral mucosa and within cheek pouches, People experience cognitive dissonance when they entertain 2 multifocal grey raised nodules in the skin, and mottled mesenteric conflicting ideas at the same time. Cognitive dissonance may and retroperitoneal masses that adhered to or displaced other viscera. cause a negative emotional state, which can lead to engagement of Livers were enlarged and pale or mottled. Histopathology was compensation mechanisms to resolve the dissonance. This report completed on 3 hamsters. Findings in the skin were multiple describes a survey which explores cognitive dissonance in laboratory epithelial neoplasms showing areas of differentiation to all segments animal veterinarians and veterinary technicians and some ways in of the hair follicle including hair bulb, inner and outer root sheath, which veterinary staff manage dissonance associated with research and accumulation of keratin and matrical shadow cells within cystic animal use. Respondents, 164 veterinarians and 145 veterinary centers. Where rupture of the neoplasm exposed keratin, a secondary technicians, were asked to rate their opinions of various statements granulomatous dermatitis was present. In abdominal tissue sections, on a scale of 1-10, where the numbers represented “strongly disagree sheets of moderately pleomorphic neoplastic round cells formed to strongly agree” or “never to always.” Statements assessed negative mesenteric masses and infiltrated the liver, kidney, GALT, and emotions (shame, powerlessness, and frustration) and compensation mesenteric lymph nodes with areas of necrosis and inflammation. mechanisms (devaluing, emotional distancing, and shifting These tumor cells had large, irregular, round-to-oval nuclei, finely responsibility). Responses were evaluated overall and compared by reticulated chromatin, and scant cytoplasm. Gross and histologic level of training (veterinarian verus veterinary technician), years of lesions were characteristic of trichoepitheliomas and lymphoma. A work experience (0-5, 6-10, >10), and species tended (large, mixed, diagnosis of transmissible lymphoma induced by hamster small species). Respondents strongly agreed that animal wellbeing polyomavirus (HaPyV) was made based on prior pathogen and animal use in research were important. Respondents reported surveillance testing at the sending institution, clinical presentation, feelings of shame, powerlessness, and frustration associated with and gross and histologic evaluation. work, but did not consistently agree to feeling powerless to initiate changes affecting animal welfare. The most frequent compensation PS118 Changing Human Behavior Towards Animals to Improve mechanism noted was shifting responsibility onto the IACUC and Laboratory Animal Welfare institutional rules. Devaluing the animals was also reported as a compensation mechanism. Responses to emotional distancing Z Brown2, J Reniers2, PV Turner*1,2 statements were divided. The survey supports the existence of cognitive dissonance associated with laboratory animal medicine. 1Global Animal Welfare, Charles River, Wilmington, MA; 2University Potential negative and positive impacts on animal welfare are of Guelph, Guelph, , Canada discussed.

Visits to research facilities in Canada and the U.S. by various regulatory and accreditation organizations commonly demonstrate gaps in routine implementation of tools, practices, and techniques to enhance the environment and care of research rodents. While part of the answer for this may lie in a need for further animal welfare science and 3Rs research, some of this is also related to resistance in

723 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019

PS120 Conflict of Interest in Research: What Is It and Why Should with a high level of national and international liaison with scientific You Care? and regulatory communities, we have been able to establish an integrated program of work aimed at reducing and ultimately ending S Pritt* severe suffering. We promote constructive dialogue between those who are involved in the use, care, and regulation of research and COI & IACUC, UT Southwestern Medical Center, Dallas, TX testing to identify practical strategies to avoid, or reduce the impact of, severe models and procedures. Our approach is well supported Conflict of interest (COI) is receiving increasing scrutiny in by the scientific community and the UK Government. Our pioneering biomedical research. Discussions about COI impact all research due initiative has so far included the organization of 2 major international to requirements found in federal regulations, scientific journals, conferences, the convening of several expert working groups, and the research contracts, grant agency guidelines, and institutional policies, production, publication, and dissemination of a range of resources, many of which originate out of concerns about industry influence including a dedicated web resource to help reduce and ultimately on academic research from over 20 years ago. Recently, the failure to end severe suffering. We highlight how the RSPCA, as a scientific disclose industry relationships in scientific manuscripts has resulted animal welfare organization, working with, rather than against, the in increased media and public attention to academic research scientific community has managed to improve implementation of the programs along with faculty consulting arrangements. Concurrently, 3Rs. Practical examples of approaches to reduce severe suffering and academic research programs have heightened institutional policies a range of resources that we have produced will be presented. regarding COI disclosures and management as well. Since several studies indicate that even small financial relationships may influence PS123 Increasing Benefit in Harm/Benefit Analysis by Robust research results, public concern about COI will only continue to Study Design intensify. We review the foundations of the current concerns with academic-industry research affiliations, which directly resulted in J Rigney* the current federal regulations on financial COI (FCOI). The most common types of COIs will be discussed and made relevant to Office Of Animal Welfare, Ethics, & Strategy, GlaxoSmithKline, animal research oversight. Differences between FCOIs and COI for Collegeville, PA IACUC review, as per Animal Welfare Act Regulations and the Public Health Service Policy requirements, will be critically examined. IACUCs work to minimize harms and ensure judicious use of Additionally, the impact of rigorous COI review, management, and animals. Harm-benefit analysis (HBA) of the experiment is usually enforcement on animal research and researchers at various institution discussed during committee review. The HBA assesses whether types (academic, industry, government) will be evaluated. the inflicted harm on the animals is outweighed by the potential benefit of the experiment. Committees generally accept the science PS121 Development of a Laboratory Animal Care Training Program justification for the procedure is valid, based on the scientist’s citing of published data or previous work. Data generated from MJ Horn*, M Holmes preclinical studies should be reproducible and translatable to the human condition, pathway, or target. If not, the benefit of the study Envigo, Indianapolis, IN may not outweigh the harm to the animals. What are the tools to select the best model and design a study that is reproducible, Laboratory animal technicians are tasked with immense translatable, and robust? Are there ways to avoid publication bias, responsibility in caring for laboratory animals that support life- that is, studies with “positive” outcomes validating a hypothesis, saving research. Attention to proper husbandry and animal care or a novel scientific discovery get published rather than those with are imperative for upholding animal welfare. Many academic uncertain or negative outcomes. Knowing this, how can we be institutions now offer training programs for animal technicians, assured that we are minimizing harm and maximizing benefit? Tools and companies exist to provide training for animal care technicians. developed in house facilitate determination of quality study design There are also several other resources available for training content, driven by preclinical-clinical continuity and translational relevance such as the AALAS Learning Library. It is also helpful for individual and pathobiology of clinical disease. The tools are prestudy review institutions to develop their own training programs based on their by a multidisciplinary Animal Model Strategy Team (AMST), and own standards of care and vivariums. We share the development employment of an Animal Model Quality Assessment (AMQA), and implementation of a training program for a large commercial followed by retrospective After Action Review (AAR) of animal institution where lessons learned may be applied to any institution. models. Additionally, applying robust study design principles, such We discuss how the training program was developed, including as randomization, blinding, and power analysis of experiments use of best practice sharing amongst facilities, creation of a training uses animals judiciously and avoids progression of projects with guidebook for 2 types of housing environments, as well as program misleading data, and nonreproducibility of experiments. Application implementation and assessment. of these tools, in conjunction with robust study design, increase benefit and minimize harm. Rapid advances in data analytics PS122 Focus On Severe Suffering: How a Scientific Animal Welfare and data sharing initiatives could further decrease the effects of Organization Can Promote Change publication bias generating advances in 3Rs principles.

E Lilley* PS124 On the CUSP: A New Option for Addressing Administrative Burden Research Animals, Royal Society for the Prevention of Cruelty to Animals, Southwater, United Kingdom A Schoenleben*1, S Thompson-Iritani2

All laboratory animal suffering is a concern, but the Royal Society for 1Office of Animal Welfare, University of Washington, Seattle, the Prevention of Cruelty to Animals (RSPCA) believes that ending WA; 2Washington National Primate Research Center, Seattle, WA ‘severe’ suffering (equivalent to some procedures within category D/E of USDA Pain and Distress Categories) should be a top priority. Under the auspices of the Federal Demonstration Partnership, the There are a number of reasons to do this: the ethical obligation to Compliance Unit Standard Procedure (CUSP) Project offers an option avoid or reduce suffering, the requirement to adhere to the 3Rs to address administrative burden at the institutional level. The goal principles, and the scientific benefits. It is widely acknowledged that of this project is to create an online resource where institutions can good quality science goes hand in hand with good welfare, and that share standard procedures used in animal care protocols with the unalleviated suffering can introduce avoidable variation and reduce broader animal welfare compliance community. A working group, the power of experiments. As a scientific animal welfare organization representing over 40 institutions, has been formed to support site

724 Abstracts of scientific papers 2019 AALAS National Meeting

design and development. The group has made significant progress both the volume and type of new PIs joining the program. By over the past year, and the site is currently in development and streamlining the PI on-boarding process, relevant information is testing. We provide an overview of the CUSP project, as well as an disseminated efficiently and a positive relationship is fostered with update on the current status and what attendees can expect moving the new investigator and the IACUC. forward. PS127 Understanding Compassion Fatigue: Associations between PS125 Reaching Research Goals through Animal Behavioral Euthanasia, Social Support, and Other Factors with Personnel Training Quality of Life

AM Ostdiek*, H Arnott, S Best, C Sandell, N Barka M LaFollette1, M Riley1, S Cloutier2, C Brady3, M O’Haire4, BN Gaskill*1 Physiological Research Laboratories, Medtronic, Coon Rapids, MN 1Animal Sciences, Purdue University, West Lafayette, IN; 2Canadian Animal research models are necessary to adequately study new Council for Animal Care, Ottowa, , Canada; 3youth development medical technologies and are required to bring new products to and agricultural education, Purdue University, West Lafayette, market. To obtain viable data, animals should be physically healthy IN; 4Comparative Pathobiology, Purdue University, West Lafayette, and housed in an environment with minimal external stressors. IN Behavioral acclimation and training can decrease stress and allow for easier data collection. Dogs, sheep, and pigs here walk on treadmills, Laboratory animal personnel may be subject to significant workplace spend time in slings, carry external equipment in saddlebags, and stress because of constantly caring for animals that may experience are acclimated to a variety of conditions. Recently we introduced pain or distress and are typically euthanized at the end of the project. clicker and target training to provide better animal welfare and easier Workplace stress is often described as compassion fatigue, which animal handling for animal care staff. Clicker training is a type of is comprised of burnout and secondary traumatic stress (a result operant conditioning which uses a distinct click to mark desired of viewing stress in others). We explored the associations between behaviors. Animals are taught to associate the click with a food treat. professional quality of life (compassion fatigue as well as the After learning the association, the sound is used to train animals opposite, compassion satisfaction) and potential risk or protective to other behaviors, such as touching their nose to a target. Target factors in laboratory animal personnel. A total of 802 laboratory training encourages animals to step onto a scale to be weighed, stand animal personnel, from the United States and Canada, completed still while being examined or data is being collected, return to their an online survey regarding professional quality of life, social kennel, and walk on a treadmill. These refinements have decreased support, euthanasia, stress/pain levels, and general interactions behavioral signs of stress in the animals when they’re being handled with laboratory animals. Personnel worked in a wide range of and provide additional cognitive enrichment. The care staff can settings (e.g., industry, universities), research types (e.g., basic, perform their duties in a safer and more efficient manner since the applied), species (e.g., nonhuman primates, mice), and roles (e.g., animals are better trained. Overall these behavioral refinements have animal care technicians, veterinarians). Data were analyzed using improved study data collection, animal welfare, and staff morale. general linear models. Our results indicate that higher self-reported levels of burnout and secondary traumatic stress were associated PS126 Setting New Principal Investigators up for Success by with less social support, enrichment provision, and control over Streamlining the On-boarding Process euthanasia (P < 0.05). Additionally, lower levels of burnout and secondary traumatic stress were associated with lower levels of K Cavanaugh*1, S Pritt2 stress or pain in the animals being cared for, the personnel’s desire for more enrichment provision, and general positive behaviors 1IACUC, UT Southwestern Medical Center, Dallas, TX; 2COI & towards laboratory animals (P < 0.05). Surprisingly, neither primary IACUC, UT Southwestern Medical Center, Dallas, TX species the personnel worked with nor frequency of euthanasia performed by the survey participant was associated with workplace New principal investigators (PIs) require guidance in both stress levels (P > 0.05). In summary, higher reported levels of social establishing their first animal protocol and understanding their support, enrichment provision, and control over euthanasia, as well responsibilities as PI. Our IACUC identified a need to inform relevant as lower levels of animal stress/pain, were associated with less parties when new PIs are being on-boarded and to establish a process compassion fatigue. Developing interventions targeting these areas to provide new PIs with targeted resources to assist with their could improve the professional quality of life of laboratory animal transition. In this large research program, there are many channels personnel. through which animal research service departments may learn of the arrival of a new PI. Once a new PI is identified, the IACUC office PS128 Impact of Euthanasia on Compassion Fatigue in Personnel is alerted and then acts as the centralized communication source to Working in Animal Research notify key personnel across departments. Standardized messaging allows on-boarding tasks between departments (e.g. granting NM Gades*1, KL Thompson2, TR Meier2, C Page3, L Mi4, M Watson2, system access, coordinating animal shipments) to occur in tandem, AM Harris5, KJ Yost5 reducing burden on administrators as well as the new PI. The IACUC office also sends each new PI a welcome message guiding them to 1Comparative Medicine, Mayo Clinic Arizona, Scottsdale, a New PI Resources webpage. This webpage was created by the AZ; 2Comparative Medicine, Mayo Clinic Rochester, Rochester, IACUC office in an effort to establish a “one-stop-shop” for new MN; 3Comparative Medicine, Mayo Clinic Florida, Jacksonville, PIs, providing relevant information on topics such as accessing the FL; 4Research Biostatistics, Mayo Clinic Arizona, Scottsdale, electronic protocol management system, writing an animal protocol, AZ; 5Health Sciences Research, Mayo Clinic Rochester, Rochester, completing required training, and obtaining animal facility access. MN Additionally, a link to OLAW’s What Investigators Need to Know About the Use of Animals is provided as an introductory resource. A cross-sectional study design was used to investigate the association Complimenting this webpage, the IACUC office also arranges a between euthanasia and compassion fatigue among employees meet and greet between each new PI, the IACUC chair/vice chair, working in animal research at a large academic medical center. The and an IACUC administrator. Meeting the new PI in person allows Professional Quality of Life Compassion Satisfaction and Fatigue for customized discussion and the ability to answer questions about questionnaire was modified to refer to animal research, and scores their specific research needs. It is also an opportunity to convey the from 3 domains (compassion satisfaction scale, burnout scale, IACUCs expectations for taking ownership of an animal protocol. and secondary traumatic stress scale) were compared between Each month, the IACUC is provided with metrics communicating individuals who euthanized animals and those who did not using

725 Vol 58, No 5 Journal of the American Association for Laboratory Animal Science September 2019 a 2-sample t-test. The overall web survey response rate was 30.8%, PS129 Murine Norovirus Mediated B Cell Depletion in Stat1 with a total of 414 responses out of 1,341 surveys. A total of 273 Knockout Mice Does Not Impair Effective Antibody Production self-reported in the survey that they euthanized animals and 73 self-reported that they did not. There was no difference (P = D Eldridge*, Mears K, C Hsu 0.10) in the compassion satisfaction T score between those who euthanized animals (mean 49.6, SD 10.0) and those who did not Comparative Medicine, University of Washington, Seattle, WA (mean 51.7, SD 10.1). There was a significant difference (P< 0.0001) in the burnout T score with greater burnout reported by those who Noroviruses have a profound global epidemiologic impact on euthanized animals (mean 51.1, SD 9.8) compared to those who human health, and it was recently shown that noroviruses can infect did not (mean 45.2, SD 9.2). Those who euthanized animals also B cells. We previously demonstrated that murine norovirus (MNV) reported significantly higher (P = 0.04) traumatic T scores (mean infection significantly reduces all stages of B cell populations in the 50.5, SD 9.96) than those who did not (mean 47.8, SD 10.0). The effect bone marrow in a signal transducer and activator of transcription sizes for score differences were further examined using Cohen’s 1 (Stat1) dependent manner as early as 7 d postinfection. Because B D to quantify the strength of the association between compassion cells are the source of antibody production, we hypothesized that satisfaction, burnout, and traumatic Stress scores and animal depletion of B cells in the bone marrow may cause an impaired euthanasia. Effect sizes were small for compassion satisfaction and ability to effectively produce antibodies when exposed to novel traumatic stress scores, -0.21 and 0.27, respectively, and medium antigens. To test this hypothesis, uninfected Stat1 knockout mice for the burnout score, 0.61. Animal research workers who euthanize (129S6/SvEv-Stat1tm1Rds) were immunized with the T-dependent animals reported significantly more burnout and traumatic stress antigen keyhole limpet hemocyanin (KLH) and then compared to than workers who do not, and the magnitude of the differences was KLH immunized mice infected with MNV-4 (n=5 mice/group), greater for burnout than for traumatic stress. Going forward, we will or uninfected Stat1 knockout mice were immunized with the analyze our survey data further to determine whether factors such T-independent antigen NP-Ficoll, and then compared to NP-Ficoll as years of employment, animal species, or method of euthanasia immunized mice infected with MNV-4 (n=5 mice/group). Serum are associated with feelings of burnout and traumatic stress among levels of antigen-specific IgG were measured by ELISA 4 wk after individuals involved in euthanasia. This information may be used to immunization. After immunization with NP-Ficoll, MNV-4 infected inform targeted interventions to mitigate these aspects of compassion Stat1 knockout mice had an average OD value of 1.04 compared to fatigue. uninfected Stat1 knockout mice with an average OD value of 0.36. After immunization with KLH, MNV-4 infected Stat1 knockout mice had an average OD value of 1.58 compared to an average OD value of 1.11 in uninfected Stat1 knockout mice. We demonstrate here that the significant depletion of B cells in the bone marrow of Stat1 knockout mice infected with MNV-4 does not result in a significant difference (P > 0.05) in serum IgG levels compared to uninfected mice immunized to either KLH or NP-Ficoll. These results suggest that B cells in MNV-4 infected mice can still mount an effective adaptive immune response to novel antigens, even in the face of diminished populations of developing B cells in the bone marrow. This effect may be due to B-cell responses outside of the bone marrow that are still able to effectively respond to antigens. Further investigation is warranted to determine the mechanisms of this effective response.

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