Newsletter

Office of Laboratory Animal Care

Volume 4, Issue 2 August 2012

What is Post-Approval Monitoring (PAM)? Dana Glass-Mattie, DVM

which means that proce- place throughout the campus dures originally written and for both teaching and re- approved get changed and search protocols and beyond, these changes occur without for those involving field being recognized by the work. Once an observation

principal investigator (PI), takes place, I will inform the Inside This Issue therefore, making the pro- PI and lab staff of any obvi- tocol noncompliant. The ous noncompliant issues and Tribromoethanol: To Use 2 goal of the PAM program is suggest methods in which to or Not To Use, That is the Post-approval monitoring, to be a positive one and help correct them which may in- Question referred to as PAM, is the identify any problems inter- clude the submission of an name commonly used for the nally before outside regula- amendment to the protocol. observation performed on tory agencies identify any. A formal report is sent to the Feral Mice in the Labora- 3 tory Animal Facility IACUC approved protocols PI by email, is included in to ensure they are executed My main job function is to the protocol file and will be as they were written. The identify protocols that have reported at the next IACUC The Animal Care and Use University of Tennessee, active animal work and go meeting. I am also available 4 Program at UTK Knoxville recently began this into the laboratory or field to to help in the initial writing program with my employ- watch the work being per- of protocols to help make ment in the role of Director formed. These observations sure they are written in a of Compliance Support include looking for any manner that makes it easier (DACS). PAM is not cur- changes in procedures, to stay compliant. The over- Focus on Biomedical 6 changes in personnel, animal Research rently a requirement of any all goal of the PAM office is regulatory body, per se, but problems, safety issues and to be a positive resource to is recommended to help work place issues that may help the University of Ten- larger institutes with a great- need to be corrected. The nessee- Knoxville reflect an er number of active proto- PAM process will begin with attitude of compliance. My cols to have a method to a contact being made to a PI office is located in Room 372 maintain compliance within of an active protocol and a in the Ellington Plant Sci- the system. Often times, time set up to observe one or ences Building and my especially with complicated all of the procedures per- phone number is 974-9074 protocols, a phenomenon formed on the protocol. for any questions or con- called ‘protocol drift’ occurs These observations will take cerns.

OLAC Office 2431 Joe Johnson Drive 336 Ellington Plant Science Knoxville, TN 37996 Phone: 865-974-5634 Fax: 865-974-5649 OLAC is published by the University of Tennessee Office of Laboratory Animal Care. UT is an EEO/AA/Title VI/Title IX/ Section 504/ADA/ADEA institution. Publication #: E18-9909-002-08 OLAC Newsletter

Tribromoethanol: To Use or Not To Use, That is the Question William A. Hill, DVM, MPH, DACLAM

sider requests for repeated dosing maceutical-grade compounds; and when adequate scientific justification specific review and approval by the is provided. IACUC. Various regimens using pharmaceutical grade anesthetics Investigators at UTK have recently have been developed for use in mice. presented findings of a study evaluat- ing repeat administration of TBE in C57BL/6NHsd mice. To our knowledge, this is the first study to thoroughly evaluate the safety and efficacy of repeat TBE administra- tion. In our study, intraperitoneal administration of TBE (500 mg/kg) Tribromoethanol (TBE), formally did not produce morbidity, mortality, available under the trade name or pathologic changes following sin- Avertin, is a non-pharmaceutical gle or repeat administration. Howev- grade anesthetic that has been used er, TBE failed to produce loss of pe- extensively for various manipulations Use of pharmaceutical grade anes- dal reflex after single administration. in laboratory rodents due to ready thetics will satisfy regulatory guid- Nine of 10 animals reached a surgical availability, lack of state and federal ance and likely result in less anes- plane of anesthesia following repeat drug regulations associated with use, thetic variability. Please contact an TBE administration, yet anesthetic and rapid anesthetic induction and OLAC veterinarian for assistance in times varied widely. Based on these recovery times.1 Despite routine use, selecting appropriate anesthetics for findings, our group urges caution in TBE use in rodents is controversial rodent species. use of TBE in C57BL/6NHsd mice due to contradictory reports regard- due to variable anesthetic effective- References: ing the compound’s efficacy and as- ness. Meyer RE, Fish RE. 2005. A review of tribromoethanol sociated pathology and mortality.2-7 anesthesia for production of genetically engineered mice and rats. Lab Animal (NY) 34:47-52. Morbidities reported with TBE use in Lieggi CC, Artwohl JE, Leszczynski JK, Rodriguez NA, mice include intestinal ileus, perito- Fickbohm BL, Fortman, JD. 2005. Efficacy and safety nitis, muscle necrosis, serositis of The UTK IACUC Guidelines for of stored and newly prepared tribromoethanol in ICR Preparation and Use of Avertin in mice. Contemp Top Lab Anim Sci 44:17-22. abdominal organs, and death.2,6,7 In Mice can be found at: Norris ML, Turner WD. 1983. An evaluation of tribro- an attempt to balance animal welfare moethanol (TBE) as an anaesthetic agent in the Mon- http://iacuc.tennessee.edu/policies/ golian gerbil (Meriones unguiculatus). Lab Anim concerns and investigator needs, avertin.shtml. 17:324-329. many institutional animal care and Papaioannou VE, Fox JG. 1993. Efficacy of tribromo- ethanol anesthesia in mice. Lab Anim Sci 43:189-192. use committees (IACUCs) have devel- Reid WC, Carmichael KP, Srinivas S, Bryant JL. 1999. oped guidelines for TBE use, includ- Pathologic changes associated with use of tribromoeth- Notwithstanding TBE effectiveness, anol (avertin) in the Sprague Dawley rat. Lab Anim Sci ing prohibiting repeat use. The UTK the National Institutes of Health Of- 49:665-667. IACUC Guidelines for Preparation fice of Laboratory Animal Welfare Tarin D, Sturdee A. 1972. Surgical anaesthesia of mice: evaluation of tribromo-ethanol, ether, halothane and and Use of Avertin in Mice can be has articulated that use of non- methoxyflurane and development of a reliable tech- nique. Lab Anim 6:79-84. found at http://iacuc.tennessee.edu/ pharmaceutical grade compounds policies/avertin.shtml. Here at UTK, Zeller W, Meier G, Bürki K, Panoussis B. 1998. Adverse such as TBE should be based on: sci- effects of tribromoethanol as used in the production of TBE is not recommended for repeat- entific necessity; no availability of transgenic mice. Lab. Anim. 32:407-413. ed use however, the IACUC will con- acceptable veterinary or human phar-

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Feral Mice in the Laboratory Animal Facility Chris Carter BS, LVT, LATg

Feral mice are highly adaptive animals of our “clean” laboratory rodents. In laboratory animal facilities employ the and frequently live in close proximity addition to posing a threat to laborato- use of live traps in all animal rooms to humans. In their search for food ry animals, wild mice have the poten- and storage areas to maximize the and shelter mice frequently find their tial to transmit zoonotic diseases to chances that the mouse will be caught way into houses, office buildings, and humans as well. before it has a chance to cause prob- even laboratory animal facilities. If all lems. the right conditions are met such as the presence of food and a warm, safe If a live mouse is caught, the OLAC office should be contacted as quickly place to nest, feral mice will inhabit as possible to ensure that a blood sam- the area and make it their permanent ple can be obtained from the mouse. home. For this reason, it is important The blood sample will be sent out and that safeguards such as general clean- a gross necropsy performed on the liness, storage of feed and bedding mouse. The information obtained above the floor, and barriers to entry from these procedures could provide are in place to prevent entry of wild an epidemiological linkage between mice. the captured feral rodents and new diseases that may arise in the current Even when the best precautions are Sometimes feral mice are visible and laboratory animal population. Pre- taken, wild mice can be very resource- they can be spotted scurrying across ventative strategies to keep feral mice ful and are known to squeeze through the floor, other times the presence of at bay are an important part of the an opening only ¼ inch in size. Wild mouse droppings is the only sign that pest control plan in a laboratory ani- mice should always be considered they are present in the facility. Mice mal facility and are a key ingredient to “dirty” animals because their health prefer walls and corners to being out impeding the spread of adventitious status is unknown and they can serve in the open, so placing a live trap agents to our rodent population. as host to a number of pathogens along the wall will result in the best which could threaten the health status chance of catching the animal. Our The Animal Care and Use Program at UTK Patricia Coan, DVM, PhD, DACLAM

Our animal care and use program in- Animal Facility (WLSLAF); The following Research and Education cludes The University of Tennessee  Jesse Harris Laboratory Animal Centers comprise the animal facilities Medical Center, Knoxville, TN; College Facility (JHLAF); throughout the state: of Education, Health and Human Sci-  University of Tennessee Medical  Dairy Research and Education Cen- ences; College of Arts and Sciences; Center, Knoxville Laboratory Ani- ter (DREC) College of Veterinary Medicine (CVM); mal Facilities (UTMCKLAF); and  East Tennessee Research and Edu- College of Agricultural Sciences and  UTK Veterinary Medical Center cation Center (ETREC) Natural Resources; and Agricultural (VMC) DAFs, which include: CVM  Greeneville Research and Education Research and Education Centers. Laboratory Animal Facility Center (GREC) The following dedicated animal facili- (VMCLAF), Joe Johnson Animal ties (DAF) comprise the core animal  Highland Rim Research and Educa- Research and Teaching Unit facilities on the UTK campus: tion Center (HRREC) (JARTU), CVM Cherokee Building A  Middle Tennessee Research and  Walters Life Sciences Laboratory (VMCCBA).

Page 3 OLAC Newsletter The Animal Care and Use Program at UTK

Education Center (MTREC) Other animals utilized at the CVM are angiotensin system and its relationship  Plateau Research and Education provided veterinary care by Small Ani- to shock and sepsis; the correlation Center (PREC) mal Clinical Sciences (SACS) veteri- between hormone replacement thera- UTK’s primary purpose is to move for- narians, Large Animal Clinical Scienc- py and vascular disease in women; ward the frontiers of human es (LACS) veterinarians, or the most small animal functional and anatomic knowledge and enrich and elevate soci- appropriate veterinary designee, as imaging of novel tracers that target ety. The mission of the animal facilities determined by the AV and IO. The tumor antigens, angiogenesis, or amy- at the University of Tennessee is to REC’s utilize the Ambulatory Service, loid-related biomarkers; and the in provide excellent service in support of members of the VMC veterinary facul- vivo study of immunoglobulin light research and teaching missions while ty or private local veterinarians who chain-related pathologies. There is a concurrently providing for the safe and have been granted designation by the strong emphasis on molecular and ethical treatment of the animals. Ex- IO and the AV to provide veterinary functional imaging and many of these ternal research funding at UTK in- care to animals at the outlaying loca- research projects make use of the state creased to more than $208.5 million in tions. -of-the-art small animal SPECT, PET FY 2011. The animal care and use pro- The decentralized evolution of the Uni- and CT imaging facility located in UT- gram at UTK is AAALAC accredited versity of Tennessee’s animal care and MCK. and has 144 primary investigators (PI) use program and the inclusion of units with 301 currently active IACUC- in the statewide REC’s have resulted in approved protocols. The Office of La- a number of separately operated ani- boratory Animal Care (OLAC) is the mal facilities. Although budget source centrally administered animal re- and management practices may vary source for UTK. somewhat among facilities, the IACUC The Institutional Official (IO) is James ensures that all practices and proce- P. Thompson, DVM, PhD. Oversight of dures are consistent with those de- the campus-wide animal care and use scribed in the Animal Welfare Regula- program is the responsibility of the tions, The Guide for the Care and Use Institutional Animal Care and Use of Laboratory Animals, The Guide for DEPARTMENT OF NUTRITION – Committee (IACUC) and the Attending the Care and Use of Agricultural Ani- JESSIE HARRIS (JH) mals in Research and Teaching, and Veterinarian (AV), Patricia N. Coan, The major areas of animal based re- university policies. The IACUC is re- DVM, PhD, DACLAM. The IACUC is search in the Department of Nutrition sponsible for semi-annual facility in- appointed by the IO and the AV re- include: obesity, diabetes, and oncolo- spections, semi-annual program re- ports to Dr. Thompson. Due to the var- gy utilizing rats and mice. Zucker fatty view, review and approval of standard ied nature of teaching and research rats are used as a genetic model of obe- operating procedures (SOPs), and re- activities and the size of the UTK Cam- sity. Athymic nude mice are used in view and approval of all teaching and pus, animal care is decentralized and xenograph experiments to study pros- research protocols involving vertebrate several satellite facilities are main- tate cancer after it has metastasized to animals. tained. The Office of Laboratory Ani- a secondary sight, such as bone. An mal Care (OLAC) is responsible for ApcMin/ mouse model is used to study research animal care at UTMCK, JH, THE GRADUATE SCHOOL OF nutrient effects on intestinal tumor- WLS, and Satellites. OLAC additionally MEDICINE – MEDICAL CENTER igenesis. provides veterinary care for some ani- KNOXVILLE (UTMCK) mals used in instructional activities at Research at the UTMCK is focused in a THE COLLEGE OF ARTS AND the College of Veterinary Medicine and variety of biomedical fields that in- SCIENCES –WALTERS LIFE SCI- for the Animal Science Department. clude: the study of the renin- ENCES (WLS)

Page 4 Volume 4, Issue 2 The Animal Care and Use Program at UTK

Biochemistry, Cellular, and Molecular and applied and basic research using members. In wildlife, teaching faculty Biology (BCMB) faculty and students laboratory, exotic, avian, wildlife, and conduct demonstrations on small use wild-type mutant and transgenic livestock species. Research emphasis mammal, amphibian, and bird han- mice to study germ cell development, areas involving animals include ad- dling to prepare students for graduate cancer, aging, cell-cycle regulation, vanced imaging, oncology, and cancer school, veterinary school, or careers in regulation of circadian rhythms and cell biology, nutrition and metabolic natural resources. Research programs responses to fungal infections. Rabbits disorders, vascular and degenerative cover amphibian diseases and conser- are used for production of polyclonal disorders, rehabilitation, and veteri- vation concerns, zoonotic disease in- antibodies. Oocytes from frogs nary public health. Teaching protocols vestigations, deer ecology and move- (Xenopus) are used for expression and include routine diagnostic and surgical ments, bird behavior and ecology, fish biochemical analysis of membrane procedures for companion animals conservation and restoration, and car- proteins. Research on vertebrate ani- and livestock. nivore ecology. mals in the Department of Ecology and Evolutionary Biology include snake THE COLLEGE OF AGRICUL- TENNESSEE AGRICULTURAL behavior, ecology, and phylogeogra- TURAL SCIENCES AND NATU- RESEARCH AND EDUCATION phy, and salamander population ge- RAL RESOURCES (CASNR) CENTERS (REC) netics and ecology. Faculty in the De- partment of Psychology study the vocal interactions of birds and effects of so- cial stress in hamsters. Those faculty members in the Department of Micro- biology who use the animal facility ag- gressively pursue research in immu- nology; virology; and microbial pathol- ogy. These investigators focus on re- search problems that use the mouse as the model animal. Typical research Animal research by Tennessee Agricul- projects include studies on: virulence The College of Agricultural Sciences tural REC scientists are focused on and population genetics of Toxoplas- and Natural Resources strives to pro- food animals and are principally di- ma gondii; cytomegaloviruses' modu- vide high-quality, experiential learning rected toward improving the efficiency lation of the immune system; lipid- opportunities for our students to pre- of food animal production systems. based virulence determinants in myco- pare them for careers in the animal Dairy and beef systems receive the ma- bacterial species; B cell responses to and wildlife industries, biotechnology, jor attention. The REC’s maintain influenza infection and vaccination; and professional programs, including three research dairy herds and four and, viral immunology and memory T Veterinary Colleges, Pharmaceutical research beef herds. Scientists use the- cell differentiation. and Medical Schools and graduate pro- se research herds to explore issues re- THE VETERINARY MEDICAL grams. To support such, the Depart- lated to food safety, animal health, re- CENTER (VMC) ment of Animal Science provides clas- production efficiency, nutrition, and Research ses covering hands-on instructional environmental impact. When called for performed laboratories involving major food ani- by specific research protocols, swine, by VMC mal (swine, bovine, ovine, and poultry) poultry, sheep, and fish are acquired in investiga- and equine species. The live animal adequate quantities and maintained in tors in- based instructional laboratories are appropriate facilities for duration of cludes clini- coordinated by Animal Science faculty specific studies. cal trials

Page 5 OLAC Newsletter

Spotlight on Models in Animal Research Tim Sparer, PhD

pesvirus called cytomegalovirus (CMV) stroke or heart attack. Human CMV has contributes to CMV dissemination and been implicated as a contributor to the pathology. Human CMV is a ubiquitous development of atherosclerotic plaques. herpesvirus with over 70% of humans We wanted to know how it was doing this. infected. In most cases the initial infec- Because immune cells are one of the initi- tion with CMV occurs in childhood with ators of plaque formation, viral and host little or no symptoms. As with all herpes- chemokines are probably responsible for viruses, after initial infection, the virus some portion of atherosclerosis. One of remains latent for the rest of their lives. the advantages of using mice is that mice Even though most people carry this virus have been generated that knock out spe- for life, it is asymptomatic unless a person cific genes allowing them to develop ath- becomes immune compromised. This erosclerosis either under a high fat diet or could occur during late stage AIDs, cancer spontaneously. These atherosclerotic therapies, or long-term suppression fol- mice will allow us to see how CMV infec- Dr. Sparer received his BS from lowing organ transplantation. Once a tion contributes to the inflammation that Northwestern University, PhD from person is immune suppressed the virus is initiates atherosclerosis. Without these Emory Medical School, and contin- reactivated and can lead to complications, mouse models the effects of CMV and its ued his training as postdoctoral fel- death or organ rejection. In order to un- role in atherosclerosis could not be discov- low at Imperial College, London, UK derstand the role that chemokines play at ered. and Stanford University Medical each stage of infection, we use the mouse More recently my lab has branched out Center. He joined the UT Microbi- model of CMV infection. One of the fea- into the field of cancer biology with a fo- ology Department in 2003. tures of CMV is its species specificity. cus on a chemokine receptor instead of This means that human CMV does not the chemokine ligand. We have found ANIMAL MODELS FOR VIRAL productively infect mice and mouse CMV that a common chemokine receptor, PATHOGENESIS, CANCER, AND does not infect humans. The only way to CXCR2, can undergo a single point muta- ATHEROSCLEROSIS understand human CMV’s lifecycle and to tion that locks it in the “ON” position. We The role of chemokines in different diseas- test potential vaccines/ treatments in vivo hypothesized that once this protein is es is the overarching focus of my lab. is to infect mice with mouse CMV. The turned on, it could contribute to cancer Chemokines are small chemoattractant mouse model for human CMV infection metastasis. After all, 90% of cancer proteins. Their secretion leads to the at- mimics many of the aspects of human deaths are due to metastasized tumor cells traction of multiple different cell types. CMV. Using this model along with recom- instead of the primary tumor so an acti- Any cell of the body that is undergoing binant mouse CMVs expressing host and vated chemokine receptor could direct stress (i.e. physical trauma, viral infection, viral chemokines, we seek to address the cancer cells to distant sites in the body etc.) will release these proteins to alert role of host and viral chemokines in viral and contribute to cancer mortality. We and recruit cells of the immune system. dissemination and pathogenesis. By un- have made cell lines that express these Think of chemokines as the “early warning covering novel roles of these chemokines mutated chemokine receptors and inject- system” of body, informing it that there is we hope to exploit its weakness for devel- ed them into mice to measure how quickly a problem or invasion from outside. We oping anti-virals that will minimize CMV the tumor grows and how well it can me- are interested in how these chemokines pathology. tastasize. Being able to measure the contribute to viral spread within the host, The mouse model for atherosclerosis al- spread of the tumor cells to distant sites in atherosclerosis (i.e. hardening of the ar- lowed us to test the role that viral and host the mouse will help us develop novel tools teries), and cancer metastasis. Mouse chemokines play in CMV-enhanced ather- for combatting metastatic cancer initiated models for studying chemokines within osclerosis. Atherosclerosis is the develop- with this chemokine receptor. Without these different disease states have been ment of fat filled plaques lining the arter- the mouse we would not be able to under- essential. ies. These plaques block the flow of blood stand the role that chemokines and their One of our projects is to study how a leading to high blood pressure but more receptors play in cancer development and chemokine produced from a common her- importantly they can rupture leading to a metastasis.

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