Journal of the American Association for Laboratory Animal Science Vol 49, No 5 Copyright 2010 September 2010 by the American Association for Laboratory Animal Science Pages 623–632

A Comprehensive Laboratory Animal Facility Response Plan

Gordon S Roble,1,2,* Naomi M Lingenhol,3 Bryan Baker,3 Amy Wilkerson,4 and Ravi J Tolwani1,3

The potential of a severe pandemic necessitates the development of an organized, rational plan for continued laboratory animal facility operation without compromise of the welfare of animals. A comprehensive laboratory animal program pandemic response plan was integrated into a university-wide plan. Preparation involved input from all levels of organizational hierarchy including the IACUC. Many contingencies and operational scenarios were considered based on the severity and duration of the . Trigger points for systematic action steps were based on the World Health Organization’s phase alert criteria. One extreme scenario requires hibernation of research operations and maintenance of reduced numbers of laboratory animal colonies for a period of up to 6 mo. This plan includes active recruitment and cross- training of volunteers for essential personnel positions, protective measures for employee and family health, logistical arrangements for delivery and storage of food and bedding, the removal of waste, and the potential for euthanasia. Strate- gies such as encouraging and subsidizing cryopreservation of unique strains were undertaken to protect valuable research assets and intellectual property. Elements of this plan were put into practice after escalation of the pandemic alerts due to influenza A (H1N1) in April 2009.

Abbreviations: RUCBC, The Rockefeller University Comparative Bioscience Center; WHO, World Health Organization.

The recent onset of a worldwide influenza A H1N1 outbreak, this time due to a novel triple among human, as well as the potential for an influenza A H5N1 pandemic, avian, viruses.19 brings to bear the possible exposure of latent weaknesses in Influenza A is the only member of theOrthomyxoviridae family traditional disaster plans of animal research facilities. Many of viruses that can infect birds, humans, and other mammals. disaster plans focus on abrupt, local disasters such as fire, Wild waterfowl serve as reservoirs for this single-stranded, earthquake, and severe storms. Although these events cause negative-sense RNA virus. Two surface antigens determine the structural damage and harm to human and animal life, they subtype specificity of influenza A. Hemagglutinin (HA or H) is a typically disrupt normal operations for a relatively short period viral envelope glycoprotein that binds to sialic acid receptors on of time. By contrast, a severe influenza pandemic could disrupt the host cell surface, permitting fusion of viral and endosomal operations for many months. Although the word ‘pandemic’ membranes and release of virion particles into the cytoplasm. invokes images of severe worldwide disease and possibly Hemagglutinin has the capability to continually mutate to avoid panic within the general population, the adjective ‘pandemic’ recognition by the host’s immune system.6 Neuraminidase indicates spread of disease and does not necessarily correlate (NA or N) allows newly formed virions to bud off from the with severity of disease. The World Health Organization (WHO) surface of the host cell. Reassortment of viral genomes between classifies a disease outbreak as a pandemic when sustained species-specific influenza strains creates new substrains with human-to-human spread of disease is present in countries of increased host range and potential pathogenicity.19 The lack at least 2 of the 6 WHO geographic regions.38 of previous immunologic exposure to a new substrain allows Many experts have posited a modern-day influenza pan- disease to freely spread within the human population, leading demic inevitable.11 Most suspected it would be from the H5N1 to pandemic illness. influenza strain rather than other subtypes such as the H1N1 Largely as a result of the anticipation of epidemics of H5N1 ‘swine flu’ of 2009.11 This assumption reflected the temporal influenza or severe acute respiratory syndrome, pandemic readi- spacing of previous .16 Three major pandemics oc- ness planning by governmental agencies and public and private curred during the 20th century; the most severe (commonly employers increased greatly during the last 10 y. Given that an known as the ‘’) happened in 1918 to 1919. This estimated 25% of all disease transmissions are believed to occur catastrophic illness arrived in 3 waves over an 18-mo period in the workplace, most national and local health organizations and killed upward of 100 million people worldwide.16 The (including the United States Department of Health and Human 1918 virus was an influenza A H1N1 subtype that arose Services and its Centers for Disease Control and Prevention through avian–human viral reassortment. In comparison, and the WHO) and local and state authorities such as the New the pandemic of 2009 resulted from another H1N1 subtype, York City Department of Health and Mental Hygiene have strongly urged employers to develop pandemic response plans that define and maintain essential functions during periods Received: 04 Dec 2009. Revision requested: 06 Jan 2010. Accepted: 28 Jan 2010. with high absentee rates.17,37 These sources provide workplace 1Tri-Institutional Training Program in Laboratory Animal Medicine and Science, 2Me- guidelines for the protection of human life, protection of assets, morial Sloan-Kettering Cancer Center, The Rockefeller University and the Weill Cornell Medical College, 3Comparative Bioscience Center, and 4Research Support, The Rockefeller and return to normal function. These recommendations assist University, New York, New York. animal facilities with disaster planning; however, they do not *Corresponding author. Email: [email protected]

623 Vol 49, No 5 Journal of the American Association for Laboratory Animal Science September 2010 provide guidance for the welfare of laboratory animals during dent’s office to an incident commander at an external agency, a pandemic event. The current pandemic of influenza A H1N1 in accordance with the overarching University disaster plan. In emphasizes the need for rational consideration when formulat- addition, a separate management hierarchy delineating essential ing and initiating an action plan. roles has been established for the RUCBC (Figure 2). To meet this challenge, The Rockefeller University Com- We also identified support structures (including housing, parative Bioscience Center (RUCBC) established an Operations transportation, and healthcare) that the University can provide Continuity Plan to outline essential services of the animal to keep essential employees onsite. Plans for facilitating an research department and developed the means to maintain extended shutdown include increasing the ability of the infor- these services in the event of a severe pandemic, most likely mation technology infrastructure to perform remote functions due to influenza. The plan modeled multiple contingencies efficiently. Plant operations personnel have been identified to based on the varying severities and durations of present and maintain and repair hardware, machinery, and systems that historic pandemics. Here we describe our 6-phase response to must remain active, even during a pandemic. a scenario during which the appearance of disease results in We also evaluated our financial and procurement procedures persistent disruption of normal work patterns for 6 mo or longer. to ensure that key financial actions (for example, running Because the early influenza 2009 outbreak was characterized payroll, paying vendors) can be conducted. The University by moderate morbidity and low mortality, full activation of a has had an emergency planning and response budget in place pandemic response plan within our facility would have been for several years. This provision has allowed planning and premature. However, we did implement limited aspects of preparation of the pandemic plan. During a severe pandemic, the Operation Continuity Plan. This effort provided valuable per diem recovery costs will cease. The Rockefeller University experience for future pandemic responses to a potentially more has anticipated the need to dramatically modify and reallocate debilitating disease. funds to cover necessary operations and commitments during an actual pandemic. Case Study Overall, the University’s primary goals during an influenza Developing an operations plan: institutional planning team. pandemic will be to 1) protect the health and safety of the Uni- After hazard analysis of pandemic flu, The Rockefeller Univer- versity community; 2) preserve and protect University resources sity established a pandemic plan working committee in 2006. and assets; 3) communicate rapidly, accurately, and frequently This committee evaluated what, in addition to the already with members of the University community; and 4) work with well-established policies and procedures outlined in the Uni- appropriate government agencies and organizations in support versity Disaster Plan, the University should do to prepare for, of and in response to their actions and efforts. respond to, and recover after a pandemic. The working com- Laboratory animal facility planning team. Animal welfare must mittee included department and resource center representatives be protected along with the health and safety of the University from Human Resources, Laboratory Safety and Environmental community. Under the care of the RUCBC are approximately Health, Finance, Hospital, Materials Management, Occupa- 25,000 mouse cages, predominantly in individually ventilated tional Health Services, Security, Plant Operations, University racks, as well as other species including large zebrafish and Residences, and the vivarium, the RUCBC. Since that initial songbird colonies and other, smaller colonies of species under evaluation, the University has taken several important steps, regulation by the US Department of Agriculture. including providing information concerning both seasonal We identified personnel with diverse expertise within the and pandemic influenza, running infection control campaigns animal facility to assist in developing a specialized plan for the during influenza season, expanding the seasonal influenza vac- RUCBC that provided guidance beyond that of the University- cination program for employees, and developing a three-level wide response plan. The charge to the planning team was to University-wide pandemic response plan that serves as guidance identify specific roles and responsibilities of animal care staff for University leadership. The University-wide response plan during a pandemic and develop plans and procedures for identifies key action steps based on local and national disease the continuity of animal facility operations in the event that prevalence, guidance from outside agencies such as the New York cessation of operations was required. Institution-wide issues City Office of Emergency Management, and to University illness including sick leave and time-off policies, expectations and rates. This template also describes the response and operational incentives for essential personnel, implementing resources to responsibilities of specific persons and groups. enable remote operations and telecommuting, and assigning An important element of the University-wide response plan is authority for shutting down operations are addressed in the the classification of events as trigger points for closing different University pandemic response plan. Plan contributors from parts of the University’s operations. The most severe response the animal facility included veterinarians, managers, and staff is the cessation of normal operations and placement of the Uni- involved in daily operations within the animal facility. Labo- versity into a state of hibernation. The committee has reviewed ratory animal facility members all have different insights into all University functions and identified those departments that day-to-day operations, and each person can provide relevant 36 can and should be shut down at different stages of a pandemic. suggestions. IACUC approval should follow development of a From this information, we identified 107 roles essential to well-delineated plan. A plan for continued IACUC functioning maintain critical facilities and resources. Of these 107 essential during a pandemic is not discussed here, given that the Office jobs, 80 must be done onsite, including roles within the depart- of Laboratory Animal Welfare provides excellent guidelines for 34 ments of security, facilities, finance, hospital, safety, information maintaining IACUC responsibilities during a pandemic. technology, housing, materials management, and animal care. Action step criteria. Our operations plan template (Figures 3 All necessary personnel were placed into a definitive chain of and 4) outlines a 2-tiered system of action steps that are based 39 command to prevent confusion during implementation of the on the WHO 8 level phase-alert classification. The first 6 WHO University-wide response plan. Figure 1 demonstrates key phases represent the identification and spread of an influenza University management changes to be implemented during a virus capable of causing pandemic human illness. Levels 5 and pandemic crisis. Responsibility is reallocated from the Presi- 6 of the WHO classification scheme correspond to declaration

624 Animal facility pandemic response plan

Figure 1. The command structure for The Rockefeller University in a disaster. This command structure is the same for all disaster types, includ- ing pandemic disease. Management command occurs from an external agency incident commander, who communicates with the University and liaisons from other agencies. This situation differs from the standard management hierarchy of The Rockefeller University, illustrated in the figure insert. of a pandemic. These phase-alert criteria serve as a real-time 1918 to 1919 influenza pandemic was category 5 (case fatality monitor of disease transmission in our plan. For increased rate greater than 2%).35 Qualification of disease severity can guidance, the plan also incorporates an internal severity index. determine the rapidity of response, in that action steps during Severity is recognized as moderate or severe; mild illness can be a moderate pandemic do not need to be executed as rapidly as treated as though due to normal seasonal influenza, with which those for a severe pandemic (Figure 4). workplaces contend yearly. The severity index is determined Plan implementation and logistics. Initial onset of pandemic from secondary features, including advisories from the local (levels 0 and 1). Our early-level (levels 0 to 1) guidelines are department of health, closures of school and public meeting derived from recommendations of the US Department of Health places, and the degree of absenteeism at the institution (Figure and Human Services and its Centers for Disease Control and 5). Severity is gauged also from the Pandemic Severity Index Prevention, the WHO, and principles of nonpharmaceutical developed by the US Department of Health and Human Services intervention. The majority of prepandemic planning occurs and its Centers for Disease Control and Prevention. This Index, during level 0, as outlined in Figure 3 and discussed later. Em- a 5-category scale, enumerates severity as case fatality rates, ployees are educated in methods to reduce disease transmission, which are the proportion of deaths among clinically ill persons.35 with emphasis on simple methods including hand washing In May 2009, the H1N1 pandemic was characterized as category with soap and water and the use of hand sanitizer. Hands- 2 (case fatality rate, 0.1% to less than 0.5%), thus qualifying as free sanitizer distributors are located throughout the facility. moderate severity in our action plan.35 In comparison, the severe Also stressed is the avoidance of touching noses, mouths, and

625 Vol 49, No 5 Journal of the American Association for Laboratory Animal Science September 2010

Figure 2. The organizational hierarchy of The Rockefeller University Comparative Bioscience Center (RUCBC). Shown in shaded boxes are the essential roles considered the minimum adequate for staffing the RUCBC during the pandemic period; the number of personnel required for each role is indicated in parentheses. This number is the same for each of the 2 rotating teams. Supervisors are expected to play an integral role in job performance and oversight. All team members will be sequestered on campus, except the veterinarian and administrative staff member who are available on an on-call basis. The need for accurate communication is exemplified by the representation of the disruption of the normal chain of command during a pandemic.

of personal protective equipment (such as N95 respirators) are recommended. has been modeled to be effective in mitigating disease spread.7 Nonpharmaceutical interventions allow near-normal levels of continuance of op- erations for as long as possible, because these measures help to maintain a healthy population of personnel to sustain essential care operations for laboratory animals. Even for these early action steps, information regarding plan implementation is disseminated from the University President’s office to the campus community by means of currently estab- lished communication systems. Notification methods include the use of The Rockefeller University general and critical infor- mation websites, ‘phone trees,’ written postings, and distributed information. In addition, we have a contract with a third-party emergency notification system that has the capability to notify as many 2500 people simultaneously through multiple sources (text message, email, and cell and landline phone calls) for each person. Once hibernation has begun, communication will con- tinue through the use of cell phones, landlines, the Internet, and person-to-person communication devices. This communication is critical, because much of the RUCBC management staff will be required to provide direction from their homes, due to the Figure 3. The Rockefeller University Comparative Bioscience Center disjoint in the management hierarchy created by University Level 0 response plan: guidelines for facility assessment and prepan- demic planning. hibernation (Figure 2). Later pandemic stages (levels 2 and 3). Further pandemic progression and increased severity of disease may sufficiently reduce the number of personnel reporting to work that modifi- eyes and staying home when sick. Employees are educated to cations in vivarium and University operations are required. In recognize early signs of illness, such as sore throat, fever, and extreme scenarios, the University may elect to cease nonessential cough. This recognition is important in minimizing spread, in research operations and continue only essential operations, such that statistics show each infected person, on average, spreads as providing basic care and support to the laboratory animals—a disease to 2 noninfected people.33 As the pandemic progresses phase of operations in our response plan described as ‘hiber- (level 2 to 3), more stringent social distancing techniques (for nation.’ For the rest of the University, hibernation includes example, avoiding hand-shaking with coworkers) and the use complete cessation of benchtop research activity in an attempt

626 Animal facility pandemic response plan

Figure 4. The RUCBC response plan in accordance with the University response plan. This 2-tiered system is guided by the WHO phase-alert criteria37 and the severity of the pandemic. Important planning or implementation steps are outlined beneath each level and degree of severity. to maintain employee health and disrupt the disease cycle. census at the time of University hibernation to allow provision Ideally, valuable research will not be lost irrevocably because of sufficient care to provide appropriate welfare of remaining researchers are alerted to the step-by-step process of placing the animals. University in the hibernation mode. Investigators should be able Recruitment, training, and support of essential personnel. to suspend their work under conditions that will facilitate the Multiple operational plans were developed for various disease reestablishment of research at University restart. In certain cases scenarios, ranging from those leading to slightly reduced staff- in which active animal research is ongoing when hibernation ing levels through to those necessitating full hibernation of the is initiated, researchers may have to consider euthanasia due research facility. Predictions for the initial stages of a severe to the inability of the RUCBC to allocate resources to manage pandemic were that staffing would be decreased to levels that and monitor these animals. would require modifications of operations, such as increasing Hibernation is not anticipated to occur unless employee the time between cage changes. If pandemic severity required absenteeism reaches 30% to 40% of the University workforce. the University to suspend research activities, essential personnel During hibernation, only essential personnel, estimated to be would be expected to perform rotating shifts of 2 wk each. To less than 10% of the normal work force, will be on campus. facilitate this situation, especially if regional shutdowns made For this and other modeled scenarios, our plan features active commuting difficult or if risk of exposure to personnel during recruitment and cross training of volunteers for these essential commutes was unacceptably high, the University will house roles. In addition, the plan includes decreasing the animal these personnel on campus by using vacant University-owned

627 Vol 49, No 5 Journal of the American Association for Laboratory Animal Science September 2010

18 people was necessary to maintain minimal function in the RUCBC (Figure 2). These calculations were based on factors such as standard metrics of productivity, defined as the number of technicians able to change 1000 rodent cages per week. Extend- ing work shifts to 12 h daily for 7 d each week, reducing cage change frequencies, and making other operational modifications are expected to increase technician productivity. Operational modifications include reducing the number of rodent cages by approximately half. The increase in shift length is expected to be the most important means of increasing work output with fewer personnel. Given the morbidity associated historic pandemics, 20,33 we expect an illness attrition of 30% to 40% of our essential personnel. This attrition rate also includes non-ill employees, such as those remaining home to care for sick family members or children dismissed from school.8 Another concern is that once the pandemic reaches severe conditions, healthy essential personnel may cease to participate, creating a personnel void. Figure 5. Considerations in determination of pandemic severity. We hope that by providing for both essential personnel and their families, the incentive plan creates sufficient impetus for and -operated apartment and hotel units and unassigned patient employees to fulfill their agreement. Members who cease to rooms. Two independent teams of personnel were viewed as participate would be replaced through extra recruited person- necessary to maintaining limited operations (Figure 2). nel. Extra personnel were recruited to replace ill members of the Participation on essential personnel teams is voluntary but 2 teams. These employees would remain at home unless their an incentive program was developed to attract volunteers. status as essential personnel was activated. Past experience with Potential incentives include increased wages while sequestered University staff performance during other disruptions, such as during the hibernation period, access to antiviral medications regional blackouts, transit strikes, and winter storms, indicates for active essential personnel and their families, vaccinations, a strong commitment by the facility staff and lends confidence and sabbatical time after the emergency. Persons volunteering that sufficient numbers of personnel will be able and willing to to fill essential roles undergo both physical and psychologic report during a pandemic. evaluation. The physical exam provides a baseline health evalu- The rotating schedule was created to minimize exposure time ation and accommodates prescription of antiviral medications to infection while accommodating the psychologic ramifications to volunteers. As seasonal and H1N1 flu vaccines become of isolation within the workplace. The ideal situation would be available, essential personnel should receive them. Fit testing a single work team that remained on campus, with no outside of and training for particulate filter respirators is arranged contact until the pandemic had receded. Because we felt this with occupational health and environmental health and safety ideal scenario would put unnecessary pressure of isolation on departments. This process fulfills employee preparedness in that work team, a rotation schedule was created. accordance with established respiratory protection programs. Providing logistical support for employees and animals is a At this time, The Rockefeller University anticipates use of daunting task. The RUCBC, like most laboratory animal facili- N95 respirators for protection against aerosol transmission of ties, operates on a just-in-time principle of supply acquisition. influenza. The literature is sparse regarding the efficiency of Normally, RUCBC receives a weekly food and bedding delivery; N95 respirators compared with surgical masks in protection the new stock is rotated as part of an existing stockpile that is against influenza spread. One recent study showed that use of sufficient for 2 wk. During a severe pandemic, manufacturing surgical masks to be within 1% of the efficacy of N95 respira- and distributing companies may lack the personnel to maintain 12 tors in preventing influenza. Limitations in the cited study a regular delivery schedule. This shortage would require the do not support complete confidence in the results. However, RUCBC to transition to maintaining larger supply quantities after combining those findings with recommendations from the on hand. Advance discussion with our essential suppliers WHO and Occupational Safety and Health Association, facilities has allowed us to establish Memoranda of Understanding to cannot be faulted for recommending the use of surgical masks facilitate the bulk delivery of appropriate materials at the time as preventative measures, particularly in the event of a shortage of request. Through these Memoranda, we are established as a 20,30 of N95 respirators. priority customer for whom deliveries are assured dependant The Rockefeller University will provide documentation to our on the capability of the supplier. Additional Memoranda were essential personnel indicating their special status during disaster generated with critical equipment companies to supply main- periods. These identification cards are available through private tenance and repair. The RUCBC will increase stores to maintain enterprises that work closely with local governments. All facili- 1-mo supplies of food and bedding stock. Long-term storage ties should consider registering essential personnel with their of materials such as bedding, food, water, and emergency municipal governments to facilitate travel and identification equipment (such as flashlights and communication devices) during disaster periods. Municipalities often provide at-cost are challenging for a facility that is already space-constrained. verification of registration in the form of specialized identifica- Adequate storage space must be allocated in vermin-proof areas, tion. Similar arrangements should be made with the appropriate according to the guidelines of The Guide for the Care and Use of officials to allow continued deliveries from outside suppliers. Laboratory Animals.10 RUCBC essential personnel are assigned specific tasks, but For sequestered personnel, arrangements for provision of cross training is provided so that essential personnel can per- nonperishable human food are in place. The University evalu- form multiple functions. We determined that a population of ated options for ready-to-eat meal items that were originally

628 Animal facility pandemic response plan

developed for military purposes but are now available from Considerations for large animals are more complex than those commercial sources, as a supplement to, and in extreme cases for rodents and other small animals. Larger animals require replacement for, meals from neighborhood commercial sources more frequent cage cleaning than do smaller animals, due to or the University’s dining services. Although these ready-to-eat the greater volume of waste produced. Facilities with sizable meal items can be stored for upward of 5 to 10 y, they generally populations of large animals must dedicate a greater propor- are not meant to be the sole source of food for more than 21 d tion of personnel to the care of these animals. Some facilities consecutively because of their typically high sodium, fat, and may consider switching bedding material from disposable bed- calorie levels. At least a 2- to 4-wk supply of ready-to-eat meal ding to durable rubber matting to reduce waste quantities and items is recommended. facilitate cleaning during this time. A temporary switch from Operational changes reducing labor needs and conserving dry-food diets to wet-food diets will ease storage and procure- supplies. For small, rack-caged animals, increasing the time ment requirements. Nonhuman primates present additional between cage changes and reducing the amount of bedding difficulties, in that many of these animals receive supplementary used per cage are 2 methods to extend the supply of bedding. fresh fruits and vegetables. Because fresh food may be difficult Our plan decreases cage-change frequency according to the to obtain during a pandemic, nonperishable treats (such as percentage of husbandry staff reporting to work. Reductions in dehydrated fruits and vegetables) can be used instead. staffing levels by 7% to 30% would require decreasing rodent The Rockefeller University has a large zebrafish colony. Many cage-changing frequency from once a week to every other week. of these animals are valuable transgenic or mutant strains. As Absentee levels beyond 30% would require further reduction part of our aquatics disaster preparation plan, redundant or in cage change frequency for a certain percentage of the rodent backup life-support equipment including water pumps and colonies. Multiple studies have been performed to examine power sources are in place. Care for zebrafish colonies is a time the effect of extending cage change intervals to 2 or 3 wk. This intensive endeavor. Essential personnel will be familiar with extension can be accomplished by modifying cage populations, these species and the associated standard operating procedure ventilation rates, and bedding types and amounts.22-24,26 Cage to prevent catastrophic loss in the unexpected event of a power changes every 3 wk require IACUC approval at The Rockefeller failure or water shutdown. University and could be instituted if bedding supplies became Population management. Food and bedding shortages and increasingly scarce. Frequent spot changes of dirty cages can decreased staffing could negatively influence animal welfare. extend the time between complete cage changes. Loss of utili- Unfortunately, in contrast to many other disasters, an evacuation ties including power and water are not anticipated during a location will not be available for animals during a pandemic. A pandemic, so ventilated caging and cage washer use should not comprehensive plan for a severe, prolonged pandemic must in- be interrupted. The Rockefeller University has guidelines for clude arrangements for the humane euthanasia of a percentage animal care in the event of utilities loss; these guidelines likely of animals in the facility to prevent potential animal suffering. extend to power loss during a pandemic. To this end, research personnel should be involved in pandemic Facilities choosing to decrease the amount of bedding disaster planning from the beginning or be informed as early in provided in each cage should consider the absorbency of the the process as possible to allow for their input. One established material, cage size, and the size and number of animals in each agreement gives each principal investigator a quota of mouse cage. Animals must be able to dig within or lay on the bed- cages that would not be euthanized in the event of a pandemic. ding in as normal a manner as possible and must be protected How investigators distribute this quota is laboratory-dependent; from temperature fluctuations. Available reviews regarding however, guidelines are provided to aid their decision (Figure 6). the absorbency and ammonia-minimizing capabilities of vari- Within this quota, each strain will be maintained at a maximum ous bedding materials should be consulted when making this of a 6-cage breeding colony. Specialized cage cards were created decision.3,22,27 In addition to ameliorating workloads, these to denote these cages. husbandry changes will reduce cage waste. Reductions in cage Critical forethought by investigators is required to determine change frequency also affect the number of cages and bottles which rodent colonies to maintain in the event of a severe, pro- sanitized weekly, reducing the number of personnel needed longed pandemic. Input from animal facility management to to support cage wash operations. Until processes such as soft the scientific staff should help guide decisions, because colony hydrothermal processing to recycle dirty bedding become com- size ultimately will be predicated on resource constraints. One monplace, cage waste will be a considerable problem, especially approach is for each laboratory to assign someone to assess if removal of waste is delayed.15 and prioritize the strains maintained by their laboratory. This Workload reduction also requires considerations of food information should be updated on a periodic basis. If labora- management. In a normal air environment of approximately tory requests for preservation of animals exceed the predicted 72 °F and 50% relative humidity, most dry laboratory animal resources to maintain these colonies, then senior institutional diets can be used as long as 6 mo after the milling date.9 A officials, such as a key scientific officer, will be asked to deter- recent study demonstrated that 2 mice in a cage will deplete mine which strains are maintained. 250 g of chow (approximate amount for wire food hoppers) In addition, the RUCBC has created guidelines for the order in in fewer than 30 d, thus accommodating topping-up of food which euthanasia will proceed (Figure 7). Information from the supplies rather than dumping hoppers and refilling them.9 If a investigator checklist is accounted for within these guidelines. topping-up technique is used, food should be removed entirely Euthanasia guidelines depend on the variety of animal species and replaced every 6 mo from the first filling of the hopper, to housed in the facility, the ability to supply adequate care to each avoid the possibility that some 6-mo-old food remains.25 This species, the true and intangible value of the strain or breed to practice will reduce food waste dramatically for facilities that research, and the ease with which it can be replaced. Within do not already use this technique. Cages will be checked daily these guidelines, early euthanasia will occur for Biosafety Level for low water levels or empty bottles. These will be replaced as 2 and 3 mice, because they require specialized training for the necessary or during cage-bottom change. husbandry staff and increased precautions for waste disposal. Similarly, training mice and mice without strain identification

629 Vol 49, No 5 Journal of the American Association for Laboratory Animal Science September 2010

proximately 333 h (or 13 d) per employee per station. For this reason, The Rockefeller University has purchased a euthanasia rack to facilitate humane euthanasia of rodents.14 These systems

provide consistent CO2 delivery for safe and efficacious eutha- nasia and significantly reduce the time required to euthanize rodents. This feature enables decreased staffing to provide bet- ter animal care to remaining colonies. In addition, this system may ameliorate the psychologic toll on employees, in that the euthanasia rack is automated and therefore less personal. An important consideration in euthanasia planning is providing a compassionate explanation of the need for euthanasia to the animal care staff because of the documented psychologic toll it imposes on many of those required to conduct the euthanasia.1 During an influenza outbreak, stringent husbandry require- ments and depopulation will protect both humans and animals. Influenza does not strike humans only; the potential exists for humans to pass disease to the animals in their care. Ferrets can catch the flu and are commonly used to study influenza due to the similar pathogenesis of disease between them and humans.18 Strains of influenza A (H1N1) have caused signs of lethargy, inappetence, and nasal discharge in ferrets.18 Other studies have demonstrated weight loss beyond acceptable IACUC parameters in infected ferrets, necessitating euthanasia.13 Pigs, Figure 6. besides giving their name to the influenza A (H1N1) of 2009, are Example of guidelines provided to laboratories to assist in 5 prioritizing animals for euthanasia. Higher-priority animals (4 > 3 > capable of acquiring the disease from people. Other influenza 2 > 1) will be euthanized later during a pandemic if necessary. These strains, such as H5N1 , can infect animals other guidelines are based on the ability of The Rockefeller University Com- than birds. Both dogs and cats have both developed clinical parative Bioscience Center to provide care and management during a signs and died following with influenza exposure to H5N1 pandemic and on the ability to repopulate postpandemic. through ingestion of infected birds.28,32 There are recent reports of household cats infected with the H1N1 virus.29 Any animals in a laboratory setting that display clinical signs consistent with influenza infection should be euthanized in an attempt to break the disease cycle within the facility Accumulation of animal carcasses and normal waste products creates considerable logistical difficulties. Disposal of waste bedding, waste food, and animal carcasses is necessary to pre- vent a public health hazard due to odor and the potential for pathogen growth. Municipal waste, cage waste, and biowaste must be considered individually, in that each may need to be removed by a different transporter dependent on facility loca- tion. The Rockefeller University has a fully permitted pathogen destructor, which allows for onsite disposal of ‘normal’ animal remains. Alternate treatment technologies, such as alkaline Figure 7. Order of euthanasia of animals within the RUCBC. The first hydrolysis tissue digestion, allow institutions to dispose of animals to be euthanized in the event of a severe pandemic would be Animal Biosafety Level 2 and 3 remains. For smaller facilities, the animals in row 1, followed by animals listed in subsequent rows as needed. Row 6 animals will not be euthanized baring extreme, un- carcass remains can be stored within freezer units for disposal predicted circumstances. when available. However, onsite freezer storage may not be fea- sible for larger facilities. Removal of these carcasses during the middle of a pandemic may prove impossible due to contractor will not be used during a pandemic period. These mice, like shutdowns; facilities must consider how carcasses are pack- vendor wild-type strains and other nontransgenic animals, aged and stored during times when onsite or offsite treatment will be available commercially after the pandemic. Animals is not available. As with other essential services and supplies that are more specialized, such as internally derived strains, (such as food, bedding, and laboratory gases), facilities should will be maintained for as long as possible if they are not part of arrange for critical and priority service guarantees from waste the specifically requested do-not-euthanize group, due to the transportation and disposal companies. potential difficulties in their rederivation postpandemic. The Rockefeller University understands that euthanasia on Mass euthanasia of rodents for colony reduction creates any scale provokes discussion. For this reason, we anticipate several logistical problems. Carbon dioxide euthanasia of the preparation of 2 sources of communication regarding our mice is used as an example here, because of its prevalence. decision. One will be an internally communicated document Many facilities have the capability to euthanize rodents on a providing explanation of our actions to the campus community, cage-by-cage basis in several locations throughout the facility. especially to those who are not involved with animal work and The Rockefeller University requires a CO exposure time of 10 2 who may not have prior knowledge of the RUCBC Operations to 15 min before opening the cage to room air to ensure high Continuity Plan. Another document will be prepared to provid- certainty of death in adult rodents.2 Euthanasia of 10,000 cages ing explanation to direct inquiries from outside sources. At this of rodents by using 5 euthanasia stations would require ap-

630 Animal facility pandemic response plan

time, The Rockefeller University does not anticipate a general 2. Artwohl J, Brown P, Corning B, Stein S. 2006. Report of the press release regarding euthanasia of animals during disaster ACLAM Task Force on Rodent Euthanasia. J Am Assoc Lab Anim response measures. Sci 45:98–105. Mitigation of research loss. The long-term cessation of a re- 3. Burn CC, Mason GJ. 2005. Absorbencies of 6 different rodent beddings: commercially advertised absorbencies are potentially search facility has even longer lasting effects on research. One misleading. Lab Anim 39:68–74. method to mitigate disaster effects is the preservation of valu- 4. Carmichael C, Westerfield M, Varga ZM. 2009. Cryopreservation able strains of animals, especially mice and fish. Embryo and and in vitro fertilization at the zebrafish international resource sperm cryopreservation are options for mouse preservation. center. Methods Mol Biol 546:45–65. Recent advances have improved the efficacy of sperm cryop- 5. Cima G. 2009. H1N1 strain in humans causes concerns over animal reservation, particularly for strains such as C57BL/6, making impacts. J Am Vet Med Assoc 234:1358–1360. this technique viable for transgenic and knockout animals.21,31 6. Ekiert DC, Bhabha G, Elsliger MA, Friesen RH, Jongeneelen M, Cryopreservation techniques are available to bank important Throsby M, Goudsmit J, Wilson IA. 2009. Antibody recognition of 4 a highly conserved influenza virus epitope. Science 324:246–251. experimental lines of fish. Immediately after drafting of the 7. Glass RJ, Glass LM, Beyeler WE, Min HJ. 2006. Targeted social pandemic response plan for the RUCBC, researchers were distancing design for pandemic influenza. Emerg Infect Dis offered murine sperm cryopreservation and embryo cryop- 12:1671–1681. reservation services at subsidized cost to encourage them to 8. Homeland Security Council. [Internet]. 2006. National strategy protect their mouse strains. Several hundred unique strains for pandemic influenza implementation plan. [Cited Oct 2009]. have been preserved and banked. A proactive, ongoing effort Available at: http://georgewbush-whitehouse.archives.gov/ to cryopreserve mouse strains as they are produced is a prudent homeland/nspi_implementation_chap01.pdf strategy to protect these valuable assets during severe pandemic 9. Huerkamp MJ, Dowdy MR. 2008. Diet replenishment for ad- libitum-fed mice housed in social groups is compatible with shelf conditions. This strategy also minimizes the potential losses due life. J Am Assoc Lab Anim Sci 47:47–50. to other catastrophic events. 10. Institute of Laboratory Animal Research. 1996. Guide for the Return to function. Once pandemic illness has subsided, care and use of laboratory animals. Washington (DC): National employees will be invited to return to work. Research activ- Academies Press. ity will take an estimated 6 to 12 mo to return to full capacity, 11. Kaiser L. 2009. An influenza smiley. Swiss Med Wkly 139:298– because animal stocks will need to be purchased or rederived. 299. All facility areas that contain no animals will be disinfected 12. Loeb M, Dafoe N, Mahony J, John M, Sarabia A, Glavin V, thoroughly prior to animal repopulation. Although these areas Webby R, Smieja M, Earn DJ, Chong S, Webb A, Walter SD. 2009. Surgical mask versus N95 respirator for preventing influenza would be cleaned during hibernation, reduced activity might among health care workers: a randomized trial. J Am Med Assoc lead to build-up of dust or other fomites. Similarly, all unused 302:1865–1871. equipment will be sanitized thoroughly or sterilized prior to use. 13. Maines TR, Jayaraman A, Belser JA, Wadford DA, Pappas C, Essential employees will be offered opportunities to meet with Zeng H, Gustin KM, Pearce MB, Viswanathan K, Shriver ZH, therapists and other medical personnel. All essential personnel Raman R, Cox NJ, Sasisekharan R, Katz JM, Tumpey TM. 2009. will be provided time-off. Transmission and pathogenesis of swine-origin 2009 A(H1N1) influenza viruses in ferrets and mice. Science 325:484–487. 14. McIntyre AR, Drummond RA, Riedel ER, Lipman NS. 2007. Conclusion Automated mouse euthanasia in an individually ventilated cag- Planning for a crisis is one of the most important under- ing system: system development and assessment. J Am Assoc Lab takings of a laboratory animal facility. Appropriate planning Anim Sci 46:65–73. protects both personnel and laboratory animals and allows for 15. Miyamoto T, Li Z, Kibushi T, Yamasaki N, Kasai N. 2008. Use continuity of animal care and welfare. The planning process is of soft hydrothermal processing to improve and recycle bedding a multistep process involving the identification of risk, essential for laboratory animals. Lab Anim 42:442–452. 16. Morens DM, Fauci AS. 2007. The 1918 influenza pandemic: in- functions and personnel to fulfill them, and operational changes sights for the 21st century. J Infect Dis 195:1018–1028. to address staffing and supply reductions. Addressing the 17. Morse SS. [Internet]. 2007. Waiting for the vaccine to arrive: psychologic needs of essential personnel is another important nonpharmaceutical interventions. [Cited Oct 2009]. Available component of the plan. The current influenza A H1N1 outbreak at: http://www.nyas.org/Publications/EBriefings/Detail. provided an opportunity for the University to test aspects of aspx?cid=e8696207-ef62-409c-a5f9-a7a5e7050e70 their response plan in real time to further strengthen its ap- 18. Munster VJ, de Wit E, van den Brand JM, Herfst S, Schrauwen plications. To date we have not advanced beyond level 1 of the EJ, Bestebroer TM, van de Vijver D, Boucher CA, Koopmans M, University response plan. Hopefully, the University will never Rimmelzwaan GF, Kuiken T, Osterhaus AD, Fouchier RA. 2009. Pathogenesis and transmission of swine-origin 2009 A(H1N1) have to enter hibernation and fully implement our pandemic influenza virus in ferrets. Science 325:481–483. response plan, but the pertinent steps are now in place, should 19. Neumann G, Noda T, Kawaoka Y. 2009. Emergence and pan- hibernation become necessary. demic potential of swine-origin H1N1 influenza virus. Nature 459:931–939. Acknowledgments 20. Occupational Safety and Health Administration. [Internet]. 2009. Pandemic influenza preparedness and response guidance for We would like to thank Dr Fred Quimby for initiating the Rockefeller healthcare workers and healthcare employers. [Cited Oct 2009]. University cryopreservation program and his helpful insights regarding Available at: http://www.osha.gov/Publications/OSHA_pan- the laboratory animal medicine pandemic plan. demic_health.pdf 21. Ostermeier GC, Wiles MV, Farley JS, Taft RA. 2008. Conserving, References distributing, and managing genetically modified mouse lines by 1. American Veterinary Medical Association. [Internet]. 2007. AVMA sperm cryopreservation. PLoS ONE 3:e2792. guidelines on euthanasia, 2007 update. [Cited Oct 2009]. Available 22. Potgieter FJ, Wilke PI. 1996. The dust content, dust generation, at: http://www.avma.org/issues/animal_welfare/euthanasia.pdf ammonia production, and absorption properties of 3 different rodent bedding types. Lab Anim 30:79–87.

631 Vol 49, No 5 Journal of the American Association for Laboratory Animal Science September 2010

23. Reeb C, Jones R, Bearg D, Bedigan H, Myers D, Paigen B. 1998. 32. Thiry E, Addie D, Belak S, Boucraut-Baralon C, Egberink H, Microenvironment in ventilated animal cages with differing venti- Frymus T, Gruffydd-Jones T, Hartmann K, Hosie MJ, Lloret A, lation rates, mice populations, and frequency of bedding changes. Lutz H, Marsilio F, Pennisi MG, Radford AD, Truyen U, Hor- Contemp Top Lab Anim Sci 37:43–49. zinek MC. 2009. H5N1 avian influenza in cats. ABCD guidelines 24. Reeb-Whitaker CK, Paigen B, Beamer WG, Bronson RT, Churchill on prevention and management. J Feline Med Surg 11:615–618. GA, Schweitzer IB, Myers DD. 2001. The impact of reduced 33. US Department of Health and Human Services. [Internet]. 2006. frequency of cage changes on the health of mice housed in venti- Pandemic influenza implementation plan—part I of II. [Cited Oct lated cages. Lab Anim 35:58–73. 2009]. Available at: http://www.hhs.gov/pandemicflu/imple- 25. Ricart Arbona RJ, Lipman N, Wolf F. 2009. Diet replenishment mentationplan/ for ad-libitum-fed mice housed in social groups is compatible with 34. US Department of Health and Human Services. [Internet]. 2006. shelf life. J Am Assoc Lab Anim Sci 48:136. PHS policy on humane care and use of laboratory animals: fre- 26. Rosenbaum MD, Vandewoude S, Johnson TE. 2009. Effects of quently asked questions. [Cited Oct 2009]. Available at: http:// cage-change frequency and bedding volume on mice and their grants.nih.gov/grants/olaw/faqs.htm#instresp_9 microenvironment. J Am Assoc Lab Anim Sci 48:763–773. 35. US Department of Health and Human Services. [Internet]. 2007. 27. Smith E, Stockwell JD, Schweitzer I, Langley SH, Smith AL. Community strategy for pandemic influenza mitigation. [Cited 2004. Evaluation of cage microenvironment of mice housed on Oct 2009]. Available at: http://pandemicflu.gov/professional/ various types of bedding materials. Contemp Top Lab Anim Sci community/commitigation.html 43:12–17. 36. Vogelweid CM. 1998. Developing emergency management plans 28. Songserm T, Amonsin A, Jam-on R, Sae-Heng N, Pariyothorn N, for university laboratory animal programs and facilities. Contemp Payungporn S, Theamboonlers A, Chutinimitkul S, Thanawon- Top Lab Anim Sci 37:52–56. gnuwech R, Poovorawan Y. 2006. Fatal avian influenza A H5N1 37. Weisfuse IB, Berg D, Gasner R, Layton M, Misener M, Zucker in a dog. Emerg Infect Dis 12:1744–1747. JR. 2006. Pandemic influenza planning in New York City. J Urban 29. Sponseller BA, Strait E, Jergens A, Trujillo J, Harmon K, Koster Health 83:351–354. L, Jenkins-Moore M, Killian M, Swenson S, Bender H, Waller K, 38. World Health Organization. [Internet]. 2009. Global alert and Miles K, Pearce T, Yoon KJ, Nara P. 2010. Influenza A pandemic response: current WHO phase of pandemic alert. [Cited Oct 2009]. (H1N1) 2009 virus infection in a domestic cat. Emerg Infect Dis Available at: http://www.who.int/csr/disease/avian_influenza/ 16:534-537. phase/en/ 30. Srinivasan A, Perl TM. 2009. Respiratory protection against in- 39. World Health Organization. [Internet]. 2009. Pandemic influenza fluenza. J Am Med Assoc 302:1903–1904. preparedness and response. [Cited Oct 2009]. Available at: http:// 31. Takeo T, Hoshii T, Kondo Y, Toyodome H, Arima H, Yamamura www.who.int/csr/disease/influenza/pipguidance2009/en/ K, Irie T, Nakagata N. 2008. Methyl-β-cyclodextrin improves index.html fertilizing ability of C57BL/6 mouse sperm after freezing and thawing by facilitating cholesterol efflux from the cells. Biol Re- prod 78:546–551.

632