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Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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4
Veterinary Care

Veterinary care is an essential part of an animal care and use Program. The primary focus of the veterinarian is to oversee the well-being and clinical care of animals used in research, testing, teaching, and production. This responsibility extends to monitoring and promoting animal well-being at all times during animal use and during all phases of the animal’s life. Well-being is determined by considering physical, physiologic, and behavioral indicators, which vary by species. The number, species, and use of animals housed in an institution may influence the complexity of the veterinary care program, but a veterinary program that offers a high quality of care and ethical standards must be provided, regardless of the number of animals or species maintained.

An adequate veterinary care program consists of assessment of animal well-being and effective management of

  • animal procurement and transportation

  • preventive medicine (including quarantine, animal biosecurity, and surveillance)

  • clinical disease, disability, or related health issues

  • protocol-associated disease, disability, and other sequelae

  • surgery and perioperative care

  • pain and distress

  • anesthesia and analgesia

  • euthanasia.

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

The veterinary care program is the responsibility of the attending veterinarian (AV), who is certified or has training or experience in laboratory animal science and medicine or is otherwise qualified in the care of the species being used. Some aspects of the veterinary care program can be conducted by persons other than a veterinarian, but a mechanism for direct and frequent communication should be established to ensure that timely and accurate information is conveyed to the responsible veterinarian about issues associated with animal health, behavior, and well-being, and that appropriate treatment or euthanasia is administered. The AV should provide guidance to investigators and all personnel involved in the care and use of animals to ensure appropriate husbandry, handling, medical treatment, immobilization, sedation, analgesia, anesthesia, and euthanasia. In addition, the AV should provide guidance and oversight to surgery programs and perioperative care involving animals.

ANIMAL PROCUREMENT AND TRANSPORTATION

Animal Procurement

All animals must be acquired lawfully, and the receiving institution should ensure that all procedures involving animal procurement are conducted in a lawful manner. Before procuring animals, the principal investigator should confirm that there are sufficient facilities and expertise to house and manage the species being acquired. Procurement of animals should be linked to the prior approval of animal use and number by the IACUC (see Chapter 2, Protocol Review). If dogs and cats are obtained from random sources, such as shelters or pounds, the animals should be inspected for tattoos or identification devices such as subcutaneous transponders (NRC 2009b); such identification might indicate that an animal was a pet, and if so, ownership should be verified. Attention should also be given to the population status of the species under consideration; the threatened or endangered status of species is updated annually by the Fish and Wildlife Service (DOI 2007). Appropriate records and other forms of documentation should be maintained for animals acquired by an institution for its investigators.

Potential vendors should be evaluated for the quality of animals they supply. As a rule, vendors of purpose-bred animals (e.g., USDA Class A dealers) regularly provide information that describes the genetic and pathogen status of their colonies or individual animals and relevant clinical history (e.g., vaccination status and anthelminthic administration). The use of purpose-bred and preconditioned animals is therefore preferable when consistent with the research, teaching, and testing objectives. In general, animals used for scientific purposes should not be obtained from pet stores or pet distributors due to the unknown or uncontrolled background of animals

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

from these sources and the potential for introducing health risks to personnel and other facility animals. Breeding colonies should be established based on need and managed according to principles of animal reduction such as cryopreservation for rodent stocks or strains (Robinson et al. 2003).

Transportation of Animals

Transportation of animals is governed by a number of US regulatory agencies and international bodies. The Animal Welfare Regulations (USDA 1985) set standards for interstate and export/import transportation of regulated species; the International Air Transport Association (IATA) updates the Live Animals Regulations annually and IATA member airlines and many countries agree to comply with these regulations to ensure the safe and humane transport of animals by air (IATA 2009). The Centers for Disease Control and Prevention and USDA enforce regulations to prevent the introduction, transmission, or spread of communicable diseases and regulate the importation of any animal or animal product capable of carrying a zoonotic disease. The US Fish and Wildlife Service regulates importation/exportation of wild vertebrate and invertebrate animals and their tissues. As the national authority arm of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES), the US Fish and Wildlife Service also regulates movement of CITES-listed species that are captive bred, including nonhuman primates (DOI 2007). Institutions should contact appropriate authorities to ensure compliance with any relevant statutes and other animal transportation requirements that must be met for animals to cross international boundaries, including those not of the country of final destination. The NRC publication Guidelines for the Humane Transportation of Research Animals provides a comprehensive review of this topic (NRC 2006); additional references on transportation of animals are available in Appendix A.

Animal transportation may be intrainstitutional, interinstitutional, or between a commercial or noncommercial source and a research facility. For wildlife, transportation may occur between the capture site and field holding facilities. Careful planning for all types of transportation should occur to ensure animal safety and well-being. The process of transportation should provide an appropriate level of animal biosecurity (see definition on page 109) while minimizing zoonotic risks, protecting against environmental extremes, avoiding overcrowding, providing for the animals’ physical, physiologic, or behavioral needs and comfort, and protecting the animals and personnel from physical trauma (Maher and Schub 2004).

Movement of animals within or between sites or institutions should be planned and coordinated by responsible and well-trained persons at the sending and receiving sites to minimize animal transit time or delays in

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

receipt. Shipping should be coordinated to ensure that animals arrive during normal business hours or, if delivery occurs outside of this time, that someone is available to receive them. Defining and delegating responsibility to the appropriate persons, who are knowledgeable about the needs of the species being shipped, will help ensure effective communication and planning of animal transport (AVMA 2002).

All animals in transit within and between institutions or jurisdictions should be accompanied by appropriate documentation to minimize delays in shipping and receipt. Documentation may include health certificates, sending and receiving institutions’ addresses and contacts, emergency procedures and veterinary contact information, and agency permits as needed.

For noncommercial sources of animals, in particular, it is important for the veterinarian or the veterinarian’s designee to review the health status and other housing and husbandry requirements before authorizing shipment of animals. This action will ensure that effective quarantine practices are implemented for incoming animals and address any special requirements needed to ensure animal well-being (Otto and Tolwani 2002). Special considerations may be necessary for transporting animals during certain phases of their life or in certain conditions, such as pregnant, perinatal, and geriatric animals; animals with preexisting medical conditions (e.g., diabetes mellitus); and animals surgically prepared by the supplier (FASS 2010).

Although ensuring animal biosecurity during transportation is always important, it is of particular importance for immunocompromised, genetically modified, and specific pathogen-free rodents (Jacoby and Lindsey 1998). For these animals, reinforced disposable shipping containers with filter-protected ventilation openings and internal food and water sources help ensure that microbial contamination does not occur during transit. Commercial vendors are experienced in animal transport and typically use dedicated transport systems and protocols to minimize microbiologic contamination. Noncommercial or interinstitutional transfer of rodents poses a higher risk of microbial contamination since the individuals involved may lack the required knowledge and animal biosecurity capabilities to maintain the animals’ health status. Risks due to in-transit microbial contamination of shipping container surfaces can be reduced by decontaminating the surfaces before placement of the containers in clean sites of animal facilities (NRC 1996, 2006). Transportation of animals in private vehicles is discouraged because of potential animal biosecurity, safety, health, and liability risks for the animals, personnel, and institution.

For aquatic species and amphibians, special considerations are required for transportation in an aqueous or sufficiently moist environment, and special attention should be given to avoiding temperature extremes for poikilotherms.

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

In all cases, appropriate loading and unloading facilities should be provided for the safe and secure transfer of animals at an institution. Facilities and procedures should be in place to help ensure that the environment at the site does not pose risks to animal well-being or personnel safety. During times of extreme temperatures animal transport may be detrimental to animal well-being and therefore may not be possible unless an appropriately heated or cooled means of transportation is available (Robertshaw 2004; Schrama et al. 1996).

PREVENTIVE MEDICINE

Disease prevention is an essential component of comprehensive veterinary medical care and biosecurity programs. Effective preventive medicine enhances the research value of animals by maintaining healthy animals and minimizing nonprotocol sources of variation associated with disease and inapparent infection, thus minimizing animal waste and potential effects on well-being. Preventive medicine programs consist of various combinations of policies, procedures, and equipment related to quarantine and stabilization and the separation of animals by species, source, and health status.

Animal Biosecurity

Animal biosecurity refers to all measures taken to identify, contain, prevent, and eradicate known or unknown infections that may cause clinical disease or alter physiologic and behavioral responses or otherwise make the animals unsuitable for research. Animal biosecurity practices should be applied to all species, but they are most important when housing large numbers of animals in intensive housing conditions (e.g., laboratory rodents). Limiting exposure of animals to infectious disease agents requires consideration of physical plant layout and operational practices. Separation of clean and soiled caging and equipment, and sometimes the associated staff, is often fundamental to success.

Animal biosecurity includes all measures to control known or unknown infections in laboratory animals.

A successful animal biosecurity program incorporates a number of elements: procedures that ensure that only animals of a desired defined health status enter the facility; personnel and materials, especially consumables, that do not serve as fomites; practices that reduce the likelihood of cross contamination if an infectious agent is inadvertently introduced; a comprehensive ongoing system for evaluating animals’ health status, including access to all animals; and containment and eradication, if desired, of

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

introduced infectious agents. Related components include procedures for evaluating and selecting appropriate animal suppliers (these may include quarantine and determination of animal health status if unknown); treatment of animals or their products at entry to minimize disease risks (e.g., surface disinfection of fish eggs); a comprehensive pest control program that may include evaluation of the health status of feral animals; procedures to ensure that all biologics administered to animals are free of contamination; and procedures for intra- and interfacility animal transport (e.g., transport of animals to laboratory and other facilities outside the animal facility can present challenges to animal biosecurity) (Balaban and Hampshire 2001). Additional details pertaining to these topics are available in the sections of Chapter 2 that deal with occupational health and safety.

Quarantine and Stabilization

Quarantine is the separation of newly received animals from those already in the facility, in a way that prevents potential spread of contaminants, until the health and possibly the microbial status of the newly received animals have been determined. Transportation can be stressful and may induce recrudescence of subclinical infections harbored by an animal.

An effective quarantine program minimizes the risk of introduction of pathogens into an established colony. The veterinary medical staff should implement procedures for evaluating the health and, if appropriate, the pathogen status of newly received animals, and the procedures should reflect acceptable veterinary medical practice and federal and state regulations applicable to zoonoses (Butler et al. 1995). Effective quarantine procedures are particularly helpful in limiting human exposure to zoonotic infections from nonhuman primates, such as mycobacterial infections, which necessitate specific guidelines for handling of these animals (Lerche et al. 2008; Roberts and Andrews 2008).

Information from suppliers about animal quality should be sufficient to enable a veterinarian to establish the length of quarantine, define the potential risks to personnel and animals in the colony, determine whether therapy is required before animals are released from quarantine, and, in the case of rodents, determine whether rederivation (cesarean or embryo transfer) is necessary to free the animals of specific pathogens. Rodents may not require quarantine if data from the vendor or provider are sufficiently current, complete, and reliable to define the health status of the incoming animals and if the potential for exposure to pathogens during transit is considered. When quarantine is indicated, animals from one shipment should be handled separately or be physically separated from animals from other shipments to preclude transfer of infectious agents between groups.

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

Depending on the health status of the colony animals and consistent with the animal biosecurity program in place, rodents or other animals being moved outside an animal facility for procedures (e.g., imaging or behavioral testing) may need to be held separately from their colony of origin until their health status is evaluated.

Regardless of whether the animals are quarantined, newly received animals should be given a period for physiologic, behavioral, and nutritional acclimation before their use (Obernier and Baldwin 2006). The length of time for acclimation will depend on the type and duration of animal transportation, the species, and the intended use of the animals. For animals not typically housed in research settings, consideration should be given to providing means to assist with their acclimation (e.g., shearing sheep before they are brought indoors). The need for an acclimation period has been demonstrated in mice, rats, guinea pigs, nonhuman primates, and goats, and time for acclimation is likely important for other species as well (Capitanio et al. 2006; Conour et al. 2006; Kagira et al. 2007; Landi et al. 1982; Prasad et al. 1978; Sanhouri et al. 1989; Tuli et al. 1995).

Separation by Health Status and Species

Physical separation of animals by species is recommended to prevent interspecies disease transmission and to eliminate the potential for anxiety and physiologic and behavioral changes due to interspecies conflict (Arndt et al. 2010). Such separation is usually accomplished by housing different species in separate rooms, but in some instances it may be possible with cubicles, laminar flow units, cages that have filtered air or separate ventilation, or isolators. It may also be acceptable to house different species in the same room—for example, two species that have a similar pathogen status and are behaviorally compatible (Pritchett-Corning et al. 2009), or aquatic species, as long as nets and other animal handling devices remain separate between systems.

In some species subclinical or latent infections can cause clinical disease if transmitted to another species. A few examples may serve as a guide in determining the need for separate housing by species:

  • Helicobacter bilis can infect rats and mice and may induce clinical disease in both species (Haines et al. 1998; Jacoby and Lindsey 1998; Maggio-Price et al. 2002).

  • As a rule, New World (South and Central American), Old World African, and Old World Asian species of nonhuman primates should be housed in separate rooms. Simian hemorrhagic fever (Renquist 1990) and simian immunodeficiency virus (Hirsch et al. 1991; Murphey-Corb et al. 1986), for example, cause only subclinical

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

infections in African species but induce clinical disease in Asian species.

  • Some species should be housed in separate rooms even though they are from the same geographic region. For example, squirrel monkeys (Saimiri sciureus) and tamarins (Saguinus oedipus) may be latently infected with herpesviruses (Herpesvirus saimiri and H. tamarinus, respectively), which could be transmitted to and cause a fatal epizootic disease in owl monkeys (Aotus trivirgatus) (Barahona et al. 1975; Hunt and Melendez 1966; Murphy et al. 1971).

Intraspecies separation may be essential when animals obtained from multiple sites or sources, either commercial or institutional, differ in pathogen status—for example, with respect to rat theilovirus in rats, mouse hepatitis virus in mice, bacterial gill disease in rainbow trout, Pasteurella multocida in rabbits, Macacine herpesvirus 1 (B virus) in macaque species, and Mycoplasma hyopneumoniae in swine.

Surveillance, Diagnosis, Treatment, and Control of Disease

All animals should be observed for signs of illness, injury, or abnormal behavior by a person trained to recognize such signs. As a rule, such observation should occur at least daily, but more frequent observations may be required, such as during postoperative recovery, when animals are ill or have a physical deficit, or when animals are approaching a study endpoint. Professional judgment should be used to ensure that the frequency and character of observations minimize risks to individual animals and do not compromise the research for which the animals are used.

Appropriate procedures should be in place for disease surveillance and diagnosis. Unexpected deaths and signs of illness, distress, or other deviations from normal in animals should be reported promptly and investigated, as necessary, to ensure appropriate and timely delivery of veterinary medical care. Animals that show signs of a contagious disease should be isolated from healthy animals. If an entire room or enclosure of animals is known or believed to be exposed to an infectious agent (e.g., Mycobacterium tuberculosis in nonhuman primates), the group should be kept intact during the process of diagnosis, treatment, and control.

Procedures for disease prevention, diagnosis, and therapy should be those currently accepted in veterinary and laboratory animal practice. Health monitoring programs also include veterinary herd/flock health programs for livestock and colony health monitoring programs for aquatic and rodent species. Access to diagnostic laboratory services facilitates veterinary medical care and can include gross and microscopic pathology, hematology, microbiology, parasitology, clinical chemistry, molecular diagnostics,

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

and serology. If a disease or infectious agent is identified in a facility or colony, the choice of therapy should be made by the veterinarian in consultation with the investigator. If the animal is to remain in the study, the selected treatment plan should be therapeutically sound and, when possible, interfere minimally with the research process.

Subclinical microbial infections (see Appendix A, Pathology, Clinical Pathology, and Parasitology) occur frequently in conventionally maintained rodents but can also occur in facilities designed and maintained for production and use of pathogen-free rodents if the microbial barrier is breached. Examples of infectious agents that can be subclinical but that may induce immunologic changes or alter physiologic, pharmacologic, or toxicologic responses are noroviruses, parvoviruses, mouse hepatitis virus, lymphocytic choriomeningitis virus, and Helicobacter spp. (Besselsen et al. 2008; Clifford and Watson 2008; NRC 1991a,b,c). Scientific objectives of a particular protocol, the consequences of infection in a specific strain of rodent, the potential for zoonotic disease, and the adverse effects that infectious agents may have on other animals or protocols in a facility should determine the characteristics of rodent health surveillance programs and strategies for keeping rodents free of specific pathogens.

The principal methods for detecting microbial infections in animal populations are serologic tests (e.g., flow cytometric bead immunoassays, immunofluorescent assays) but other methods, such as DNA analysis using polymerase chain reaction (PCR), microbial culture, clinical chemistry (e.g., lactate dehydrogenase virus), histopathology, and other validated emerging technologies, can also be used to make or confirm a diagnosis.

Transplantable tumors, hybridomas, cell lines, blood products, and other biologic materials can be sources of both murine and human viruses that can contaminate rodents or pose risks to laboratory personnel (Nicklas et al. 1993); rapid and effective assays are available to monitor microbiologic contamination and should be considered before introducing such material into animals (Peterson 2008).

Because health monitoring programs are dependent on the size and complexity of the Program, the species involved, and the institutional research focus, it is beyond the scope of the Guide to go into details about health monitoring programs for all species; additional references are in Appendix A (under Disease Surveillance, Diagnosis, and Treatment; Pathology, Clinical Pathology, and Parasitology; and Species-Specific References).

CLINICAL CARE AND MANAGEMENT

Healthy, well-cared-for animals are a prerequisite for good-quality animal-based science. The structure of the veterinary care program, including the number of qualified veterinarians, should be appropriate to fulfill the

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

program’s requirements, which will vary by institution, species used, and the nature of the animal use. To be effective in providing clinical care, the veterinarian should be familiar with the species and various uses of animals in the institutional research, teaching, testing, or production programs and have access to medical and experimental treatment records.

Medical Management

There should be a timely and accurate method for communication of any abnormalities in or concerns about animal health, behavior, and well-being to the veterinarian or the veterinarian’s designee. The responsibility for communicating these concerns rests with all those involved with animal care and use. Reports should be triaged to ensure that animals most in need receive priority attention, and the veterinarian or veterinarian’s designee should perform an objective assessment of the animal(s) to determine an appropriate course of action.

Well-planned experiments with clearly delineated scientific and humane endpoints will help to ensure that a contingency plan is in place for problems that may arise during the study (see Chapter 2, Experimental and Humane Endpoints). For animals on research protocols, the veterinarian or veterinarian’s designee should make every effort to discuss any problems with the principal investigator or project director to jointly determine the most appropriate course of treatment or action. Standard operating procedures (SOPs) may be developed for recurrent health conditions to expedite treatment. Recurrent or significant problems involving experimental animal health should be communicated to the IACUC, and all treatments and outcomes should be documented (USDA 1997).

Emergency Care

Procedures must be in place to provide for emergency veterinary care both during and outside of regularly scheduled hours. Such procedures must enable animal care and research staff to make timely reports of animal injury, illness, or death. A veterinarian or the veterinarian’s designee must be available to expeditiously assess the animal’s condition, treat the animal, investigate an unexpected death, or advise on euthanasia. In the case of a pressing health problem, if the responsible person (e.g., investigator) is not available or if the investigator and veterinary staff cannot reach consensus on treatment, the veterinarian must have the authority, delegated by senior administration (see Chapter 2, Institutional Official and Attending Veterinarian) and the IACUC, to treat the animal, remove it from the experiment, institute appropriate measures to relieve severe pain or distress, or perform euthanasia if necessary.

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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Recordkeeping

Medical records are a key element of the veterinary care program and are considered critical for documenting animal well-being as well as tracking animal care and use at a facility. A veterinarian should be involved in establishing, reviewing, and overseeing medical and animal use records (Field et al. 2007; Suckow and Doerning 2007). All those involved in animal care and use must comply with federal laws and regulations regarding human and veterinary drugs and treatments. Drug records and storage procedures should be reviewed during facility inspections.

SURGERY

Successful surgical outcomes require appropriate attention to presurgical planning, personnel training, anesthesia, aseptic and surgical technique, assessment of animal well-being, appropriate use of analgesics, and animal physiologic status during all phases of a protocol involving surgery and postoperative care (see Appendix A, Anesthesia, Pain, and Surgery). The individual impact of those factors will vary according to the complexity of procedures involved and the species of animal used. A team approach to a surgical project often increases the likelihood of a successful outcome by providing input from persons with different expertise (Brown and Schofield 1994; Brown et al. 1993).

Surgical outcomes should be continually and thoroughly assessed to ensure that appropriate procedures are followed and timely corrective changes are instituted. Modification of standard techniques may be required (for instance, in aquatic or field surgery), but should not compromise the well-being of the animals. In the event of modification, close assessment of outcomes may have to incorporate criteria other than clinical morbidity and mortality. Such assessments rely on continuing communication among technical staff, investigators, veterinarians, and the IACUC.

Training

Researchers conducting surgical procedures must have appropriate training to ensure that good surgical technique is practiced—that is, asepsis, gentle tissue handling, minimal dissection of tissue, appropriate use of instruments, effective hemostasis, and correct use of suture materials and patterns (Brown et al. 1993; Heon et al. 2006). Training may have to be tailored to accommodate the wide range of educational backgrounds frequently encountered in research settings. For example, persons trained in human surgery may need training in interspecies variations in anatomy, physiology, the effects of anesthetic and analgesic drugs, and/or postoperative care requirements.

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

Technical staff performing rodent surgery may have had little formal training in surgical techniques and asepsis and may require general surgical training as well as training for the specific techniques they are expected to perform (Stevens and Dey 2007).

Training guidelines for research surgery commensurate with an individual’s background are available (ASR 2009) to assist institutions in developing appropriate training programs. The IACUC, together with the AV, is responsible for determining that personnel performing surgical procedures are appropriately qualified and trained in the procedures (Anderson 2007).

Presurgical Planning

Presurgical planning should include input from all members of the surgical team (e.g., the surgeon, anesthetist, veterinarian, surgical technicians, animal care staff, and investigator). The surgical plan should identify personnel, their roles and training needs, and equipment and supplies required for the procedures planned (Cunliffe-Beamer 1993); the location and nature of the facilities in which the procedures will be conducted; and perioperative animal health assessment and care (Brown and Schofield 1994). A veterinarian should be involved in discussions of the selection of anesthetic agents and doses as well as the plan for perioperative analgesic use. If a nonsterile part of an animal, such as the gastrointestinal tract, is to be surgically exposed or if a procedure is likely to cause immunosuppression, preoperative antibiotics may be appropriate (Klement et al. 1987); however, the routine use of antibiotics should never be considered a replacement for proper aseptic surgical techniques.

Presurgical planning should specify the requirements for postsurgical monitoring, care, and recordkeeping, including the personnel who will perform these duties. The investigator and veterinarian share responsibility for ensuring that postsurgical care is appropriate.

Surgical Facilities

Unless an exception is specifically justified as an essential component of the research protocol and approved by the IACUC, aseptic surgery should be conducted in dedicated facilities or spaces. When determining the appropriate location for a surgical procedure (either a dedicated operating room/suite or an area that provides separation from other activities), the choice may depend on the species, the nature of the procedure (major, minor, or emergency), and the potential for physical impairment or post-operative complications, such as infection. Most bacteria are carried on airborne particles or fomites, so surgical facilities should be maintained and operated in a manner that ensures cleanliness and minimizes unnecessary

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

traffic (AORN 2006; Bartley 1993). If it is necessary to use an operating room for other purposes, it is imperative that the room be returned to an appropriate level of hygiene before its use for major survival surgery.

Generally, agricultural animals maintained for biomedical research should undergo surgery with techniques and in facilities compatible with the guidelines set forth in this section. However, some minor and emergency procedures commonly performed in clinical veterinary practice and in commercial agricultural settings may take place under field conditions. Even when conducted in an agricultural setting, however, these procedures require the use of appropriate aseptic technique, sedatives, analgesics, anesthetics, and conditions commensurate with the risk to the animal’s health and well-being.

Surgical Procedures

Surgical procedures are categorized as major or minor and, in the laboratory setting, can be further divided into survival and nonsurvival. As a general guideline, major survival surgery (e.g., laparotomy, thoracotomy, joint replacement, and limb amputation) penetrates and exposes a body cavity, produces substantial impairment of physical or physiologic functions, or involves extensive tissue dissection or transection (Brown et al. 1993). Minor survival surgery does not expose a body cavity and causes little or no physical impairment; this category includes wound suturing, peripheral vessel cannulation, percutaneous biopsy, routine agricultural animal procedures such as castration, and most procedures routinely done on an “outpatient” basis in veterinary clinical practice. Animals recovering from these minor procedures typically do not show significant signs of post-operative pain, have minimal complications, and return to normal function in a relatively short time. When attempting to categorize a particular surgical procedure, the following should be considered: the potential for pain and other postoperative complications; the nature of the procedure as well as the size and location of the incision(s); the duration of the procedure; and the species, health status, and age of the animal.

Laparoscopic surgeries and some procedures associated with neuroscience research (e.g., craniotomy, neurectomy) may be classified as major or minor surgery depending on their impact on the animal (Devitt et al. 2005; Hancock et al. 2005; NRC 2003; Perret-Gentil et al. 1999, 2000). For example, laparoscopic techniques with minimal associated trauma and sequelae (e.g., avian sexing and oocyte collection) could be considered minor, whereas others (e.g., hepatic lobectomy and cholecystectomy) should be considered major. Although minor laparoscopic procedures are often performed on an “outpatient” basis, appropriate aseptic technique, instruments, anesthesia, and analgesia are necessary. Whether a laparoscopic procedure is deemed

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

major or minor should be evaluated on a case-by-case basis by the veterinarian and IACUC.

Emergency situations sometimes require immediate surgical attention under less than ideal conditions. For example, if an animal maintained outdoors needs surgical attention, movement to a surgical facility might be impractical or pose an unacceptable risk to the animal. Such situations often require more intensive aftercare and may pose a greater risk of postoperative complications. The appropriate course of action requires veterinary medical judgment.

In nonsurvival surgery, an animal is euthanized before recovery from anesthesia. It may not be necessary to follow all the techniques outlined in this section if nonsurvival surgery is performed but, at a minimum, the surgical site should be clipped, the surgeon should wear gloves, and the instruments and surrounding area should be clean (Slattum et al. 1991). For nonsurvival procedures of extended duration, attention to aseptic technique may be more important in order to ensure stability of the model and a successful outcome.

Aseptic Technique

Aseptic technique is used to reduce microbial contamination to the lowest possible practical level (Mangram et al. 1999). No procedure, piece of equipment, or germicide alone can achieve that objective (Schonholtz 1976): aseptic technique requires the input and cooperation of everyone who enters the surgery area (Belkin 1992; McWilliams 1976). The contribution and importance of each practice varies with the procedure. Regardless of the species, aseptic technique includes preparation of the patient, such as hair or feather removal and disinfection of the operative site (Hofmann 1979); preparation of the surgeon, such as the provision of appropriate surgical attire, face masks, and sterile surgical gloves (Chamberlain and Houang 1984; Pereira et al. 1990; Schonholtz 1976); sterilization of instruments, supplies, and implanted materials (Bernal et al. 2009; Kagan 1992b); and the use of operative techniques to reduce the likelihood of infection (Ayliffe 1991; Kagan 1992a; Lovaglio and Lawson 1995; Ritter and Marmion 1987; Schofield 1994; Whyte 1988).

While the species of animal may influence the manner in which principles of aseptic technique are achieved (Brown 1994; Cunliffe-Beamer 1983; Gentry and French 1994), inadequate or improper technique may lead to subclinical infections that can cause adverse physiologic and behavioral responses (Beamer 1972; Bradfield et al. 1992; Cunliffe-Beamer 1990; Waynforth 1980, 1987) affecting surgical success, animal well-being, and research results (Cooper et al. 2000). General principles of aseptic technique should be followed for all survival surgical procedures (ACLAM 2001).

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
×

Specific sterilization methods should be selected on the basis of the physical characteristics of the materials to be sterilized (Callahan et al. 1995; Schofield 1994) and sterilization indicators should be used to validate that materials have been properly sterilized (Berg 1993). Autoclaving and plasma and gas sterilization are effective methods most commonly used to sterilize instruments and materials. Alternative methods, used primarily for rodent surgery, include liquid chemical sterilants and dry heat sterilization. Liquid chemical sterilants should be used with appropriate contact times and instruments should be rinsed with sterile water or saline before use. Bead or dry heat sterilizers are an effective and convenient means of rapidly sterilizing the working surfaces of surgical instruments but care should be taken to ensure that the instrument surfaces have cooled sufficiently before touching animal tissues to minimize the risk of burns. Alcohol is neither a sterilant nor a high-level disinfectant (Rutala 1990) but may be acceptable for some procedures if prolonged contact times are used (Huerkamp 2002).

Intraoperative Monitoring

Careful monitoring and timely attention to problems increase the likelihood of a successful surgical outcome (Kuhlman 2008). Monitoring includes routine evaluation of anesthetic depth and physiologic functions and conditions, such as body temperature, cardiac and respiratory rates and pattern (Flegal et al. 2009), and blood pressure (Kuhlman 2008), and should be appropriately documented. Use of balanced anesthesia, including the addition of an intraoperative analgesic agent, can help minimize physiologic fluctuations during surgery. Maintenance of normal body temperature minimizes cardiovascular and respiratory disturbances caused by anesthetic agents (Dardai and Heavner 1987; Flegal et al. 2009; Fox et al. 2008), and is of particular importance in small animals where the high ratio of surface area to body weight may easily lead to hypothermia. Fluid replacement may be a necessary component of intraoperative therapy depending on the duration and nature of the procedure. For aquatic species (including amphibians), care should be taken to keep the skin surfaces moist and minimize drying during surgical procedures.

Postoperative Care

An important component of postsurgical care is observation of the animal and intervention as necessary during recovery from anesthesia and surgery (Haskins and Eisele 1997). The intensity of monitoring will vary with the species and the procedure and may be greater during the immediate anesthetic recovery period. During this period, animals should be in a clean, dry, and comfortable area where they can be observed frequently by

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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trained personnel. Particular attention should be given to thermoregulation, cardiovascular and respiratory function, electrolyte and fluid balance, and management of postoperative pain or discomfort. Additional care may be warranted, including long-term administration of parenteral fluids, analgesics, and other drugs, as well as care of surgical incisions. Appropriate medical records should also be maintained.

After recovery from anesthesia, monitoring is often less intense but should include attention to basic biologic functions of intake and elimination and to behavioral signs of postoperative pain, monitoring for postsurgical infections, monitoring of the surgical incision site for dehiscence, bandaging as appropriate, and timely removal of skin sutures, clips, or staples (UFAW 1989).

PAIN AND DISTRESS

An integral component of veterinary medical care is prevention or alleviation of pain associated with procedural and surgical protocols. Pain is a complex experience that typically results from stimuli that damage or have the potential to damage tissue; such stimuli prompt withdrawal and evasive action. The ability to experience and respond to pain is widespread in the animal kingdom and extends beyond vertebrates (Sherwin 2001).

Pain is a stressor and, if not relieved, can lead to unacceptable levels of stress and distress in animals. Furthermore, unrelieved pain may lead to “wind-up,” a phenomenon in which central pain sensitization results in a pain response to otherwise nonpainful stimuli (allodynia; Joshi and Ogunnaike 2005). For these reasons, the proper use of anesthetics and analgesics in research animals is an ethical and scientific imperative. Recognition and Alleviation of Pain in Laboratory Animals (NRC 2009a) is an excellent source of information about the basis and control of distress and pain (see also Appendix A, Anesthesia, Pain, and Surgery).

Fundamental to the relief of pain in animals is the ability to recognize its clinical signs in specific species (Bateson 1991; Carstens and Moberg 2000; Hawkins 2002; Holton et al. 1998; Hughes and Lang 1983; Karas et al. 2008; Martini et al. 2000; Roughan and Flecknell 2000, 2003, 2004; Sneddon 2006). Species vary in their response to pain (Baumans et al. 1994; Kohn et al. 2007; Morton et al. 2005; Viñuela-Fernández et al. 2007), and criteria for assessing pain in various species differ. The U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training (see Appendix B) state that in general, unless the contrary is known or established, it should be considered that procedures that cause pain in humans may also cause pain in other animals (IRAC 1985).

Certain species-specific behavioral manifestations are used as indicators of pain or distress—for example, vocalization (dogs), depression

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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(all), anorexia (all), rapid or labored respiration (rodents, birds, fish), lack of grooming (mammals and birds), increased aggression (mammals and birds), periocular and nasal porphyrin discharge (rodents), abnormal appearance or posture (all), and immobility (all) (NRC 2008, 2009a). However, some species may mask signs of pain until they are quite severe (NRC 2009a). It is therefore essential that personnel caring for and using animals be trained in species-specific and individual clinical, behavioral, physiologic, and biochemical indicators of well-being (Dubner 1987; Karas 2002; Murrell and Johnson 2006; Rose 2002; Stoskopf 1994; Valverde and Gunkel 2005).

Distress may be defined as an aversive state in which an animal fails to cope or adjust to various stressors with which it is presented. But distress may not induce an immediate and observable pathologic or behavioral alteration, making it difficult to monitor and evaluate the animal’s state when it is present. Both the duration and intensity of the state are important considerations when trying to prioritize attention to and treatment of animal distress. For example, an injection requiring brief immobilization may produce acute stress lasting only seconds, while long-term individual housing of a social species in a metabolic cage may produce chronic distress. Implementation of clear, appropriate, and humane experimental endpoints for animals, combined with close observation during invasive periods of experimentation, will assist in minimizing distress experienced by animals used in research, teaching, testing, and production. Recognition and Alleviation of Distress in Laboratory Animals (NRC 2008) is a resource with important information about distress in experimental animals.

ANESTHESIA AND ANALGESIA

The selection of appropriate analgesics and anesthetics should reflect professional veterinary judgment as to which best meets clinical and humane requirements as well as the needs of the research protocol. The selection depends on many factors, such as the species, age, and strain or stock of the animal, the type and degree of pain, the likely effects of particular agents on specific organ systems, the nature and length of the surgical or pain-inducing procedure, and the safety of the agent, particularly if a physiologic deficit is induced by a surgical or other experimental procedure (Kona-Boun et al. 2005).

Preemptive analgesia (the administration of preoperative and intraoperative analgesia) enhances intraoperative patient stability and optimizes postoperative care and well-being by reducing postoperative pain (Coderre et al. 1993; Hedenqvist et al. 2000). Analgesia may be achieved through timely enteral or parenteral administration of analgesic agents as well as by blocking nociceptive signaling via local anesthetics (e.g., bupivacaine).

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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Alleviation of chronic pain may be more challenging than postprocedural pain; commercially available opiate slow-release transdermal patches or implantable analgesic-containing osmotic minipumps may be useful for such relief. Because of wide individual variation in response to analgesics, regardless of the initial plan for pain relief, animals should be closely monitored during and after painful procedures and should receive additional drugs, as needed, to ensure appropriate analgesic management (Karas et al. 2008; Paul-Murphy et al. 2004). Nonpharmacologic control of pain may be effective and should not be overlooked as an element of postprocedural or perioperative care for research animals (NRC 2009a; Spinelli 1990). Appropriate nursing support may include a quiet, darkened recovery or resting place, timely wound or bandage maintenance, increased ambient warmth and a soft resting surface, rehydration with oral or parenteral fluids, and a return to normal feeding through the use of highly palatable foods or treats.

Most anesthetics cause a dose-dependent depression of physiologic homeostasis and the changes can vary considerably with different agents. The level of consciousness, degree of antinociception (lack of response to noxious stimuli), and status of the cardiovascular, respiratory, musculoskeletal, and thermoregulatory systems should all be used to assess the adequacy of the anesthetic regimen. Interpretation and appropriate response to the various parameters measured require training and experience with the anesthetic regimen and the species. Loss of consciousness occurs at a light plane of anesthesia, before antinociception, and is sufficient for purposes of restraint or minor, less invasive procedures, but painful stimuli can induce a return to consciousness. Antinociception occurs at a surgical plane of anesthesia and must be ascertained before surgery. Individual animal responses vary widely and a single physiologic or nociceptive reflex response may not be adequate for assessing the surgical plane or level of analgesia (Mason and Brown 1997).

For anesthesia delivery, precision vaporizers and monitoring equipment (e.g., pulse oximeter for determining arterial blood oxygen saturation levels) increase the safety and choices of anesthetic agents for use in rodents and other small species. For injectable anesthestic protocols, specific reversal agents can minimize the incidence of some side effects related to prolonged recovery and recumbency. Guidelines for the selection and proper use of analgesic and anesthetic drugs should be developed and periodically reviewed and updated as standards and techniques are refined. Agents that provide anesthesia and analgesia must be used before their expiration dates and should be acquired, stored, their use recorded, and disposed of legally and safely.

Some classes of drugs such as sedatives, anxiolytics, and neuromuscular blocking agents may not provide analgesia but may be useful when

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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used in combination with appropriate analgesics and anesthetics to provide balanced anesthesia and to minimize stress associated with perioperative procedures. Neuromuscular blocking agents (e.g., pancuronium) are sometimes used to paralyze skeletal muscles during surgery in which general anesthetics have been administered (Klein 1987); because this paralysis eliminates many signs and reflexes used to assess anesthetic depth, autonomic nervous system changes (e.g., sudden changes in heart rate and blood pressure) can be indicators of pain related to an inadequate depth of anesthesia. It is imperative that any proposed use of neuromuscular blocking drugs be carefully evaluated by the veterinarian and IACUC to ensure the well-being of the animal. Acute stress is believed to be a consequence of paralysis in a conscious state and it is known that humans, if conscious, can experience distress when paralyzed with these drugs (NRC 2008; Van Sluyters and Oberdorfer 1991). If paralyzing agents are to be used, the appropriate amount of anesthetic should first be defined on the basis of results of a similar procedure using the anesthetic without a blocking agent (NRC 2003, 2008, 2009a).

EUTHANASIA

Euthanasia is the act of humanely killing animals by methods that induce rapid unconsciousness and death without pain or distress. Unless a deviation is justified for scientific or medical reasons, methods should be consistent with the AVMA Guidelines on Euthanasia (AVMA 2007 or later editions). In evaluating the appropriateness of methods, some of the criteria that should be considered are ability to induce loss of consciousness and death with no or only momentary pain, distress, or anxiety; reliability; irreversibility; time required to induce unconsciousness; appropriateness for the species and age of the animal; compatibility with research objectives; and the safety of and emotional effect on personnel.

Euthanasia may be planned and necessary at the end of a protocol or as a means to relieve pain or distress that cannot be alleviated by analgesics, sedatives, or other treatments. Criteria for euthanasia include protocol-specific endpoints (such as degree of a physical or behavioral deficit or tumor size) that will enable a prompt decision by the veterinarian and the investigator to ensure that the endpoint is humane and, whenever possible, the scientific objective of the protocol is achieved (see Chapter 2).

Standardized methods of euthanasia that are predictable and controllable should be developed and approved by the AV and IACUC. Euthanasia should be carried out in a manner that avoids animal distress. Automated systems for controlled and staged delivery of inhalantsmay offer advantages for species killed frequently or in large numbers, such as rodents (McIntyre et al. 2007). Special consideration should be given to euthanasia of fetuses

Suggested Citation:"4 Veterinary Care." National Research Council. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. doi: 10.17226/12910.
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and larval life forms depending on species and gestational age (Artwohl et al. 2006).

The selection of specific agents and methods for euthanasia will depend on the species involved, the animal’s age, and the objectives of the protocol. Generally, chemical agents (e.g., barbiturates, nonexplosive inhalant anesthetics) are preferable to physical methods (e.g., cervical dislocation, decapitation, use of a penetrating captive bolt); however, scientific considerations may preclude the use of chemical agents for some protocols.

Although carbon dioxide (CO2) is a commonly used method for rodent euthanasia, there is ongoing controversy about its aversive characteristics as an inhalant euthanasia agent. This is an area of active research (Conlee et al. 2005; Danneman et al. 1997; Hackbarth et al. 2000; Kirkden et al. 2008; Leach et al. 2002; Niel et al. 2008) and further study is needed to optimize the methods for CO2 euthanasia in rodents (Hawkins et al. 2006). The acceptability of CO2 as a euthanasia agent for small rodents should be evaluated as new data become available. Furthermore, because neonatal rodents are resistant to the hypoxia-inducing effects of CO2 and require longer exposure times to the agent (Artwohl et al. 2006), alternative methods should be considered (e.g., injection with chemical agents, cervical dislocation, or decapitation; Klaunberg et al. 2004; Pritchett-Corning 2009).

It is essential that euthanasia be performed by personnel skilled in methods for the species in question and in a professional and compassionate manner. Special attention is required to ensure proficiency when a physical method of euthanasia is used. Death must be confirmed by personnel trained to recognize cessation of vital signs in the species being euthanized. A secondary method of euthanasia (e.g., thoracotomy or exsanguination) can be also used to ensure death. All methods of euthanasia should be reviewed and approved by the veterinarian and IACUC.

Euthanizing animals is psychologically difficult for some animal care, veterinary, and research personnel, particularly if they perform euthanasia repetitively or are emotionally attached to the animals being euthanized (Arluke 1990; NRC 2008; Rollin 1986; Wolfle 1985). When delegating euthanasia responsibilities, supervisors should be sensitive to this issue.

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A respected resource for decades, the Guide for the Care and Use of Laboratory Animals has been updated by a committee of experts, taking into consideration input from the scientific and laboratory animal communities and the public at large. The Guide incorporates new scientific information on common laboratory animals, including aquatic species, and includes extensive references. It is organized around major components of animal use:

  • Key concepts of animal care and use. The Guide sets the framework for the humane care and use of laboratory animals.
  • Animal care and use program. The Guide discusses the concept of a broad Program of Animal Care and Use, including roles and responsibilities of the Institutional Official, Attending Veterinarian and the Institutional Animal Care and Use Committee.
  • Animal environment, husbandry, and management. A chapter on this topic is now divided into sections on terrestrial and aquatic animals and provides recommendations for housing and environment, husbandry, behavioral and population management, and more.
  • Veterinary care. The Guide discusses veterinary care and the responsibilities of the Attending Veterinarian. It includes recommendations on animal procurement and transportation, preventive medicine (including animal biosecurity), and clinical care and management. The Guide addresses distress and pain recognition and relief, and issues surrounding euthanasia.
  • Physical plant. The Guide identifies design issues, providing construction guidelines for functional areas; considerations such as drainage, vibration and noise control, and environmental monitoring; and specialized facilities for animal housing and research needs.

The Guide for the Care and Use of Laboratory Animals provides a framework for the judgments required in the management of animal facilities. This updated and expanded resource of proven value will be important to scientists and researchers, veterinarians, animal care personnel, facilities managers, institutional administrators, policy makers involved in research issues, and animal welfare advocates.

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