1. Chapter Eight Laboratory Animal Environment

2.  If viewing this in PowerPoint, use the icon to run the show (bottom left of screen).  Mac users go to “Slide Show > View Show” in menu bar  Click on the Audio icon: when it appears on the left of the slide to hear the narration.  From “File > Print” in the menu bar, choose “notes pages”, “slides 3 per page” or “outline view” for taking notes as you listen and watch the presentation.  Start your own notebook with a 3 ring binder, for later study! ALAT Presentations Study Tips

3. Environment, Equipment, & Hygiene  Standards, equipment & procedures  safety & potentially hazardous problems  Animal facility commonly located in building or wing separated from labs & offices.  Facility = animal rooms, cage wash, offices, locker rooms, storage, surgery suites & other rooms.  Environment (temperature, humidity, noise & air exchange) evaluated for level & consistency.  Animals outdoors need protection from heat, cold & other environmental extremes.

4. (Image) Animal Facility Corridor

5. Animal Rooms  Conventional = no special precautions to prevent introduction of disease  Feed, groom & vaccinate against diseases  don’t keep in special room to isolate  Larger animals in conventional cages & rooms  Barrier = special cages & procedures practiced  A barrier = cage, a room or even entire facility  Temperature, humidity, ventilation, light, noise & other variables in room affect environment.  Direct effect on behavior & health which impacts data.  The Guide provides table of recommended temperature levels for common laboratory species.

6. Temperature & Humidity  Too hot or too cold can induce stress in animals.  In rodent breeding room, ~ 72°F suitable for neonates  Newly-hatched chicks or hairless rodents, & post- operative recovery = warmer end of range.  Newly-hatched chicks & very young pigs may require an extra heat source.  Temp > 80°F, adult mice may suffer heat stress.  Humidity = amount of moisture in air.  Range of relative humidity is 30–70%.  Low humidity = ringtail & respiratory problems.  High humidity = respiratory problems & food spoilage.

7. Ventilation  Ventilation helps eliminate noxious odors.  ammonia = by-product of breakdown of urine  The Guide: 10 -15 exchanges of fresh air/hour.  Recirculated air filtered to remove contaminants.  Room is under + or – pressure.  + pressure rooms = > pressure than surroundings  Opening a door causes air to flow out toward the hallway.  Surgical suites, barrier rooms under + pressure.  - pressure rooms = < pressure than surroundings  Opening a door causes air to flow into the room.  Quarantine and conditioning & rooms to contain hazardous agents under – pressure.  Check air pressure regularly to be sure correct.

8. Lighting  Lighting evenly distributed & bright enough  Change = stress & affects breeding efficiency.  Turn lights on or off at prescribed times.  Check lights on automatic timers.  Long exposure to bright light = problem for albino.  Research may call for specific light/dark cycles.  Natural light varies & interferes w/ controlled light.  Some systems permit changing light intensity.  Lower level used to maintain animals.  Level temporarily increased for room cleaning & maintenance.

9. Noise  Certain rodents, cats, guinea pigs & rabbits react negatively to noises.  Rabbits may jump & injure self, rodents may not breed.  Noise may alter hormone levels.  Technicians should avoid making loud noises (shouting or banging cage doors).  Radio may be played to provide low-level background noise, reduces startle response.  Speech in comforting tones to noise-sensitive animals may calm them.  House these animals away from noisy animals, activities & busy areas.

10. Animal Room Construction  Ideal animal room allows easy cleaning.  Non-slip, waterproof floors & drain covers prevent debris from going into sewer.  Floors are sloped toward drains.  Walls and ceilings made of sealed material  waterproof & easy to clean  surfaces free of cracks, chipped paint & vermin-proof  moisture-proof & vermin-proof recessed fixtures & outlets  Doors self-closing, with a view port, kick plate, locks & recessed handles.  Dimensions are a minimum of 42 inches X 84 inches.

11. Barrier & Containment  More sophisticated rooms  Designed to prevent entry of disease or disease in- side room from getting out.  Shower and wear special protective clothing.  Cages, bedding, food & water sterilized before entering room to prevent introduction of disease.  SPF = animals do not have certain diseases.  Barrier rooms have standard design features, but they have a filtered air supply.  Air passes through a HEPA filter to remove dust & particles from air.

12. Hallways and Traffic Patterns  High traffic areas & require durable construction.  Bumper guards on walls and corners help protect surfaces from becoming chipped and marred.  Recommended hallway width of 6–8 feet.  Double-door airlocks at end of hallways help reduce noise, odors & possible contamination.  Doors that open inward prevent collisions.  Fixtures recessed into wall facilitate equipment movement.

13. (Image) Animal Facility Hallway

14. Hallways and Traffic Patterns II  Single hallway w/ all rooms off same hallway = clean & dirty cages through same area.  In clean/dirty system animal room has 2 doors:  1 - entering from clean hallway  2 - leading into the dirty hallway  Prevent dirty from contaminating clean  Enter room from clean, exit by dirty hallway.  Disinfect shipping boxes prior to entering facility.  Disinfect or autoclave supplies before entering barrier.  To work, staff must abide by design requirements.

15. Facility Security  Vandalism & theft add to cost of research by destroying years of effort & wasting animal lives.  Door locks, computerized card entry system  After opening security doors, check that they have been relocked.  Never give unauthorized people keys, access cards & door combinations  Find out who stranger is by asking, “How may I help you?”  Advise supervisors immediately of presence of unauthorized people.

16. Cage Washing Rooms  Cage washing activities are noisy.  Locate areas away from animal areas & offices.  Ventilate areas well to remove steam & odors.  Seal all electrical outlets, light fixtures, walls, floors & ceilings to allow proper cleaning.  prevent water & moisture from penetrating

17. Quarantine, Conditioning & Isolation  Quarantine area separates incoming animals from animals already in the facility.  During quarantine, personnel perform health status evaluations & monitor for disease.  While in quarantine area, animals adjust to new surroundings, referred to as conditioning.  Can be constructed as specialized cubicles  each cubicle equipped w/ negative-pressure air supply  Isolation for animals known or suspected of carrying disease  Isolation rooms operate on negative pressure air supply.

18. Feed and Bedding Storage  Must be vermin-proof & dry  Feed & bedding stored on shelves or pallets, away from walls.  Cool temp and low humidity avoid spoilage.  Separate storage areas designated for clean equipment.  Do not store disinfectants & chemicals w/ clean equipment, feed or bedding.  Hallways, corridors & animal rooms should not be used as storage areas.  Items obstruct traffic flow, accumulate dirt and can be a safety hazard.

19.  Personnel Areas: offices, lunch rooms, showers & locker rooms  Lunch & office areas are only place in which eating & other break activities are permitted.  Procedure rooms - investigators & technicians perform experimental or therapeutic procedures  Performing procedures in animal room may alarm other animals.  Contain examining tables for diagnostic or experimental work & some storage space. Personnel & Procedure Areas

20. Emergency Back-up Power  Without ventilation, air conditioning or heat = no fresh air in the facility.  Causes temperature and ammonia levels to rise  Prolonged high levels of ammonia or carbon dioxide can have serious effects on animal health.  Loss of negative air pressure is a serious problem in quarantine and containment areas.

21. Hazardous Agents  Use of materials or procedures that present a physical, chemical, biological or radiological hazard  Document potential problems & precautions before any research involving hazardous agents.  Signs & notices of hazard type must be posted where hazardous agents are being used.  Contaminated animals or wastes disposed of in areas bearing biohazard symbol.  ALWAYS ASK FIRST & proceed only when confident proper procedures are being followed.

22. Radioisotope Hazards  Radioisotopes emit low levels of radiation, which makes them valuable in studies of processes such as digestion.  Radioisotopes are usually dangerous only if contacted directly; some require stringent precautions & safety measures. Radiation monitoring badge records exposure and is worn by staff working with radioisotopes. Institutions using radioactive materials must conform to federal NRC guidelines.

23. Pathogen Hazards  Infectious bacteria, viruses, fungi or parasites that pose a threat to humans and animals  Some can cause disease in humans and may be present in research animals that are not SPF.  Pathogens are utilized for research studies.  i.e. developing tuberculosis vaccine  CDC publishes guidelines for handling organisms & infected animals.  Guidelines outline biosafety levels depending on degree of danger to humans.  Biosafety levels graded from 1 to 4.  BSL 1 is least & BSL 4 most hazardous. Diagram of HIV virus

24. Mutagen, Carcinogens & Toxin Hazards  Mutagens cause changes in chromosomes.  Induce occurrence of mutations.  Carcinogens can induce cancer directly.  Dilute formaldehyde (formalin) tissue preservative.  Toxins are poisonous substances.  i.e. formaldehyde and gas anesthetics  Toxins can also be produced by bacterial, plant or animal cells.  Organism that causes tetanus produces a toxin which is responsible for symptoms.

25. Hazard Containment  Following features help contain hazards:  + and - air pressure differences  Filtration of exhaust air  Airlocks and pass-through autoclaves  Ultraviolet light to destroy organisms on surfaces  Change and shower rooms  Intercom systems between areas  Treatment of contaminated sewage  Back-flow check valves in main water supply lines  Separation between containment & utility systems  SOP for each specific biohazard under study  Covers proper use of PPE

26. Dead Animal Storage & Disposal  Place in plastic bags & either freeze or refrigerate until destroyed or removed.  Plastic or metal cans used for storing carcasses should be leak proof & have tight-fitting lids.  Store only dead animals in these units  Incineration is safe & sanitary means of disposal. If animal is found dead, notify supervisor who will notify veterinarian & PI. May perform necropsy & take certain tissues from animal before disposal. Clearly label carcass & place in refrigerator until a necropsy can be performed.

27. Caging - The Microenvironment  Animals must be able to move freely & have access to clean food and water.  SOPs describe housing standards & care.  Adhering to space recommendations allows animals to have enough room to make normal postural adjustments.  Too many animals or cages that are too small causes stress.  affect physiology, behavior & research data  Crowding is both inhumane & against the law.  USDA’s AWA & The Guide provide specific guidelines.

28. (Image) Various Types of Caging

29. Bedding  Bedding = an absorbent in waste collection pans & material in direct contact with animals.  Cage design determines what type of bedding is used; direct or indirect bedding. Too muchToo littleJust right

30. Bedding II  Characteristics of good bedding material are:  No ideal material for all species in all applications.  ground corn cob, wood shavings, compressed paper & straw  All bedding products create a certain amount of dust & can clog spray jets in cage washers.

31. Additional Reading Biosafety in Microbiological and Biomedical Laboratories. HHS Publication No. (CDC) 93- 8395. 3rd. Edition US Government Printing Office, Washington, D.C. 1993. Guide for the Care and Use of Laboratory Animals. National Academy Press, 1996.