Animal Health Maintenance LAT Chapter 8

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The Daily Link Note subtle changes in animal behavior, physical appearance, or daily activity. Early discovery of an abnormality in an animal may reduce animal suffering and colony disease problems and prevent the compromise of an experiment. Contagious viral infection a threat in colony animals. “Herd Health” management Barrier system protection against a serious disease outbreak. Any signs of disease, the entire colony may be affected. If treatment is expensive or disease extensive, more feasible to depopulate (euthanize) room or colony and disinfect the facility.

Animal Acquisition Centralized = better price, transport, housing and health. Animals requests consider: source, strain, sex, age, weight, pathogen status, number and transportation. Do not change suppliers until study is completed. Vendor animal health survey - sample subjected to diagnostic procedures to determine health status. Use correct terminology to describe pathogen status. SPF or VAF may mean different things. Larger animals screening - worming and vaccination schedules, health history, and USDA paperwork.

Transportation AWA & USDA standards = length of time shipment held prior to departure minimum and maximum temperature transport facilities size, ventilation and construction of containers how many animals may be transported in a cage air exchange and temperature in the animal cargo space maximum length of time without food or water how often checked while in transit avoiding rough handling health certificates Shipping containers: conventional = cardboard, SPF and germfree = filtered air vents Document shipment conditions in vehicle and condition of animals on arrival.

Receiving Receiving person must be aware of animals expected and instructions from the investigator on how they are to be housed. Know orders placed, which vendor, what ordered, arrival, and purchase ID - required by federal inspectors. Check specifications of order. Cage cards and records Physical examinations Taconic (right) and Harlan (left) shipping containers

Receiving (continued) Disinfect containers prior to opening. Aseptically remove from containers and place in decontaminated cages in a pathogen-free area. May collect % for diagnostic confirmation of health status. Conditioning: > 48 hours required for rodents Animals may become immunosuppressed and dehydrated during shipment. If exposed to a disease, more likely to become ill. Better to discover disease in quarantine, than in existing colony. Ideal quarantine in separate rooms or isolation cubicles, by vendor and arrival date or at least be separated by species.

Shipping Do This! Link to TAC web site: Click on any of the items enclosed by a red outline and an explanation will pop-up. (Very cool Taconic!) Protect from impact, offer ventilation and heat, food and water and keep animals clean and dry. Display address, contact and special instructions. Ship early in the week in order to avoid weekend delays. Check with state veterinarian or the USDA prior. Species of animal determines shipping requirements. Shipping of birds and cold-blooded animals = unique problems. Size and other animals in the group = # animals housed. Different sizes should not be shipped in the same container. Age must be considered; very young should not be shipped in a container with older animals unless they are suckling littermates with their dam.

Quarantine & Conditioning Quarantine = observed for signs of disease For larger animals, includes treatment for internal or external parasites, tattooing, checking tuberculosis, and treating diseases. Conditioning = or the period during which animals are becoming adjusted to their new environment. Work in quarantine only after work with all other healthy animals. Negative room air pressure so undesirable microorganisms pass directly into exhaust system. Uniform change commonly required after leaving.

Quality Assurance & Monitoring Health monitoring is to ensure validity of experimental data Identify and prevent viral, bacterial, and parasitic diseases. Sentinel animal monitoring Direct or indirect exposure Diagnostic procedures are performed at scheduled intervals. Environmental monitoring = changes in the environment amount, age, quality, and palatability of food and water temperature and humidity ventilation effectiveness light cycle lengths noise level type of bedding material presence of pheromones and chemical disinfectants

Effects of Disease on Research Mouse Hepatitis Virus (MHV): high mortality in newborn mice; alters study of the immune system Sendai Virus: bronchitis and pneumonia; profound influence on the immune system Mycoplasmosis: highly contagious, progressive, inflammatory disease; affects respiratory, reproductive & immune systems Pasteurellosis: Pasteurella multocida, “snuffles” in rabbits; can infect respiratory, reproductive system & middle ear Oftentimes only way to completely eradicate disease from a colony is to euthanize entire colony, disinfect area and start over with new animals. The time and expense of this kind of drastic procedure can be disastrous to research programs.

Microbial Status Conventional: under standard conditions, number and types of microbes are not specifically known Specific Pathogen Free (SPF): describes only those microorganisms they do not contain, term SPF alone says nothing about which are present Gnotobiotic: defined microbial flora Axenic: completely free of all detectable microorganisms and parasites Caesarean-Derived or Caesarean-Origin: mammals delivered by surgical means Barrier-Reared or Barrier-Sustained: raised in a controlled microbial barrier, which prevents the entrance of pathogenic organisms

Isolator Styles

Equipment and Supplies Isolator: rigid type - stainless steel or plexiglass & flexible type - clear plastic film Individual Micro-Isolator™: shoebox-type plastic cages that have a hard plastic lid with a special filter Barrier is at the cage level. One disadvantage is that air exchange may be reduced. Ventilated cage racks: supply HEPA filtered air Supply a constant flow of fresh air to the animals, which reduces ammonia and carbon dioxide levels inside the cage. Provide either positive or negative pressure inside the cage/ Sterilized food, water and bedding done by packaging food and water in a supply cylinder (unless irradiated)/ Air is sterilized by passing it through special HEPA filters.

Ventilated Cage Racks Thoren Caging Ventilated Rack Lab Products Ventilated Rack

Conventional Animals Wear gloves and either a clean lab coat or clothing worn only inside facility. Food and water free from pathogens but not sterile. Equipment and cages sanitized to kill pathogens. Relatively inexpensive to produce and maintain. Research not type that is affected by the microbes or the minor response variations caused by those microbes. Useful in experiments measuring rate drug is absorbed across skin. Not in experiments in which fine differences in immune system’s response to bacteria are measured.

Specific Pathogen Free Defined by specific microbes they lack. Safeguards prevent them from being contaminated. Access to the rooms housing these animals is usually restricted. Shower before entering and/or wear special clothing. Supplies are usually sterilized. Common bacteria and viruses may still be in the environment without affecting SPF status. Monitoring includes tests for pathogens the animal should be free from. Investigations of a disease caused by a specific pathogen require animals free of that disease. Research involving diseases known to be influenced by the presence of specific pathogens.

Gnotobiotic and Axenic Produced by surgically removing young from mother’s sterile uterus just prior to birth. Gnotobiotic animals are associated with bacteria, which enhance digestion but do not cause disease. Food has > amounts of vitamins and other nutrients. Vitamins A, C, K, thiamine, and pyroxidine are partially destroyed by the heat of sterilization process. Special nutritional needs, because the bacteria that help produce vitamins in the intestines normally are missing. Sensitive immunological and biochemical studies are typical of the types of research that require the use of axenic or gnotobiotic animals.

Additional Reading 1. Foster, H.L. “Gnotobiology.” In: The Laboratory Rat, Volume II. Baker, H.L. et al., eds. Academic Press, Orlando, FL, 1980. 2. Small, J.D. “Rodent and Lagomorph Health Surveillance and Quality Assurance.” In: Laboratory Animal Medicine, Fox, J.G., Cohen, B.J., and Loew, F.M., eds. Academic Press, Orlando, FL, 1984.