LATG: A Review of The Guide for the Care and Use of Laboratory Animals

1. LATG:A Review ofThe Guide for the Care and Use of Laboratory Animals Created by Marc S. Hulin, DVM, Dipl. ACLAM

2. History of The Guide • First published in 1963; 6th revision • Current revision was written by ILAR • Purpose: assist institutions in caring for and using animals in ways judged to be scientifically, technically, and humanely appropriate • 6th edition was supported by NIH, USDA, and Dept. VA

3. Introduction • Laboratory Animal: any vertebrate animal used in research, teaching or testing • Guide endorses: 9 US Government principles, AWA, PHS policy, other federal and state laws

4. Evaluation Criteria • Performance standards preferred over Engineering standards • performance: define an outcome in detail and provide criteria for assessing the outcome, but don’t limit methods to achieve outcome • “Must”: considered imperative • “Should”: strong recommendation

5. I. Institutional Policies and Responsibilities • IACUC membership: • DVM with lab animal experience • 1 practicing scientist with research experience with animals • 1 public member (non-affiliated) • Physical restraint: • use of manual or mechanical means to limit animal’s movement for examination, collection of samples, or drug administration

6. Physical Restraint • Prolonged restraint should be avoided unless it is essential for research objectives and approved by IACUC • Guidelines: • not to be considered normal methods of housing • restraint period should be minimum time • animal should be trained • regular observations • veterinary care provided if lesions or illness

7. Multiple Major Surgical Procedures • Major surgery: • penetrates and exposes a body cavity or produces substantial impairment of physical or physiologic function • Multiple major surgeries on a single animal are discouraged unless: • scientifically justified • approved by the IACUC • related components of a research project • conserve scare animal resources

8. Food or Fluid Restriction • If experimental protocols require restriction at least minimal quantities of food and fluid should be available • Restriction for research purposes should be scientifically justified • Highly preferred food or fluid as positive reinforcement, instead of restriction, is recommended

9. Veterinary Care • Adequate veterinary care must be provided, including access to all animals for evaluation of their health and well-being • Veterinarian must provide investigators with advice on use of sedatives, analgesics, or anesthetics in animals

10. Personnel Qualification & Training • Personnel caring for animals should be appropriately trained and the institution should provide for formal or on-the-job training to facilitate effective implementation of the program and humane care and use of animals • Occupational Health and Safety program must be part of the overall animal care and use program

11. Preventative Medicine for Personnel • Development and Implementation of a program for medical evaluation should involve input from trained health professionals • Cercopithecine Herpesvirus 1 (Herpes B virus): • personnel who work with Macaques should have access to and be instructed in the use of bite and scratch emergency kits

12. Animal Environment, Housing, Management • Proper housing and management: • Animal well-being • Quality of research data • Health and safety of personnel • Animals should be housed with goal of maximizing species-specific behaviors and minimizing stress induced behavior

13. Micro vs. Macroenvironment • Microenvironment: • physical environment immediately surrounding it, i.e. primary enclosure, food, wire bar lid, sipper tube. • Macroenvironment: • the physical environment of the secondary enclosure, such as a room, barn, or an outdoor habitat • Linked by ventilation between primary and secondary enclosure

14. Primary Enclosures • Should be constructed with materials that balance the needs of the animal with the ability to provide sanitation • Solid-bottom caging, with bedding, is recommended for rodents

16. Space Recommendations • Based on professional judgment and experience • Thigmotactic: rodent species benefit more from wall space (stay along the walls in cages) • Animal must have enough space to turn around and express normal postural adjustments • Social animals should be housed in pairs or groups

17. Space Recommendations • Cage height: • Mice: 5 inches • Rats: 7 inches • Hamsters: 6 inches • Guinea pigs: 7 inches • Rabbits: 14 inches • Cats: 24 inches

20. Space Recommendations • Floor space: • Dogs: • <15 kg: 8.o sq. ft. • 15-30 kg: 12.0 sq. ft. • > 30 kg: 24.0 sq. ft. • Monkeys: • Groups 1-7 (including baboons) • Apes (Pongidae): Groups 1-3 • Brachiating species cage height should be when fully extended, swing from cage ceiling without feet touching the floor

23. Temperature and Humidity • Regulation of body temperature within normal variation is necessary for the well-being of lab animals • Relative humidity should be controlled to the acceptable range of 30-70%

24. Dry Bulb Temperatures • Mouse, Rat, Hamster, Gerbil, Guinea Pig: • 64-790 F • Rabbit: • 61-720 F • Cat, Dog, NHP: • 64-840 F • Farm animals & Poultry: • 61-810 F

25. Ventilation • Supply adequate oxygen; remove thermal loads caused by animals and equipment • Use of computer modeling for designing facility ventilation (computational fluid dynamics) • 10-15 fresh-air changes per hours for secondary enclosures is general standard • Recycled vs. Non-recycled air

26. Illumination • Light can affect the physiology, morphology, and behavior of various animals • Lighting should provide sufficient illumination: • for well-being of the animals • allow good housekeeping practices • Time-controlled lighting system should be used to ensure a regular diurnal cycle

27. Illumination • Light levels of 325 lux (30 ft-candles) about 1.0m above the floor appear to be sufficient for animal care and do not cause signs of phototoxic retinopathy in albino rats • Light at cage level for animal susceptible to phototoxic retinopathy should be between 130-325 lux

28. Noise • Noisy animals (dogs, swine, etc.) should be housed away from quieter animals, such as rodents, rabbits, and cats • Noise >85 dB can have both auditory and non-auditory effects: • eosinopenia • increased adrenal weights in rodents • reduced fertility in rodents • increased blood pressure in NHP

29. Behavioral Management • Structural Environment: • primary enclosure- cage complexities, cage furniture, manipulanda • Social Environment: • physical contact and communication with conspecifics (members of same species) • Activity: • animals should have opportunities to exhibit species-typical activity patterns

30. Husbandry • Food: • unused, opened bags of food should be stored in vermin-proof containers • exposures to temp. > 700 F and humidity extremes hastens deterioration • dry lab animal diets stored for 6 months after manufacture • Vitamin C diets- 3 months shelf-life • Stabilized forms of Vitamin C extend shelf life

31. Bedding • Untreated softwood shavings and chips are contraindicated may affect animals’ metabolism • Cedar shavings are not recommended, because microsomal enzymes and cytotoxicity • Soiled bedding should be removed and replaced with fresh materials as often as is necessary to keep the animals clean and dry

32. Cleaning and Disinfection of Primary Enclosures • If animal waste is to be removed by flushing-at least once daily (animals kept dry during flushing) • enclosures and accessories (tops) should be sanitized at least once every 2 weeks • Solid-bottom caging, bottles, and sipper tubes usually require sanitization at least once a week

34. Cleaning and Disinfection of Primary Enclosures • Effective disinfection can be achieved with wash & rinse water at 143-1800 F • Traditional 1800 F for rinse water refers to the water in the tank or sprayer manifold • A regularly scheduled and documented pest control and monitoring should be implemented

36. Emergency, Weekend, and Holiday Care • Animals should be cared for by qualified personnel every day, including weekends and holidays • Emergency veterinary care should be available after work hrs., weekends, and holidays • A disaster plan that takes into account both personnel and animals should be prepared as part of the overall safety plan for the animal facility

37. Identification and Records • Toe-clipping, for identification of small rodents: • only when no other method is feasible • performed only on altricial rodents • Clinical records for individual animals: • valuable for dogs, cats, NHP • contain pertinent clinical and diagnostic info. • dates of vaccinations, surgery, experimental use

38. Veterinary Medical Care • Adequate veterinary care consists of effective programs for: • Preventative Medicine • Surveillance, Diagnosis, Treatment, and control of disease including Zoonosis • Management of protocol associated disease • Anesthesia and Analgesia • Surgery and Postsurgical care • Assessment of Animal Well-Being • Euthanasia

39. Veterinary Medical Care • A veterinary care program is the responsibility of the attending veterinarian, who is certified or has training or experience in lab animal science and medicine • The veterinarian must provide guidance to investigators and all personnel involved in the care and use of animals

40. Animal Procurement & Transportation • All animals must be acquired lawfully • Dogs and cats from Class B dealers should be carefully inspected for special identification markers • Importation of NHP is regulated by PHS with specific guidelines for Tuberculin testing

41. Quarantine, Stabilization, and Separation • Effective quarantine program minimizes the chance for introduction pathogens into an established colony • Veterinary Medical staff should have procedures for evaluating the health and pathogen status of newly received animals

42. Quarantine, Stabilization, and Separation • Effective quarantine program for NHP: • limit exposure of humans to zoonotic infections • filoviral and mycobacterial infections in NHP have necessitated specific handling guidelines (CDC) • Newly received animals should be given a period for physiologic, psychologic, and nutritional stabilization before their use.

43. Quarantine, Stabilization, and Separation • Physical separation of animals by species is recommended to prevent interspecies disease transmission and conflict • separate rooms • cubicles • laminar-flow units • cages that have filtered air or separate ventilation • isolators

44. Quarantine, Stabilization, and Separation • Examples of need for separate housing by species: • Bordetella bronchiseptica in rabbits- severe disease in guinea pigs • Simian Hemorrhagic Fever and SIV separate New World, Old World African, and Old World Asian • Squirrel monkey latently infected with Herpesvirus tamarinus which is fatal to Owl Monkeys

45. Surveillance, Diagnosis, Etc. • All animals should be observed for signs of illness, injury, or abnormal behavior daily • Unexpected deaths and signs of illness should be reported promptly for appropriate veterinary medical care • The choice of medication or therapy should be made by the veterinarian in consultation with the investigator

46. Surveillance, Diagnosis, Etc. • Infectious agents that affect research: • Sendai, KRV, MHV, LCMV, and Mycoplasma pulmonis • The principal method for detecting viral infections is serologic testing • Transplantable tumors, hybridomas, cell lines, and other biologic materials should be tested for murine viruses • MAP (mouse antibody production) test, RAP, HAP used for monitoring for viral contamination

47. Surgery • Appropriate attention to presurgical planning, personnel training, aseptic and surgical technique • Use of antibiotics should never be considered as a replacement for aseptic procedures • PHS policy and AWA place responsibility with the IACUC for determining that personnel are qualified and trained for surgery

48. Surgery • Major survival surgery: • penetrates and exposes a body cavity or produces substantial impairment of physical or physiologic function • laparotomy, thoracotomy, craniotomy, joint replacement, limb amputation

49. Surgery • Minor survival surgery: • does not expose a body cavity and causes little or no physical impairment • wound suturing, peripheral-vessel cannulation, castration, prolapse repair, skin biopsy • most procedures routinely done on an “outpatient” basis in veterinary clinical practice

50. Surgery • Nonsurvival surgery: • animal is euthanized before recovery from anesthesia • does not require aseptic surgery • surgical site should be clipped, wear gloves, and clean instruments • Skin sutures, wound clips, or staples: • must be removed in timely manner • Veterinary medical standard: 10-14 days