Environment of animals and working in lab animal unit

Transcription

1 ESS 09 Environment of animals and working in lab animal unit Satu Mering PhD National Laboratory Animal Center University of Kuopio Genetics + Environment and genetics Environment Microbiology Nutrition Procedures => Phenotype Response Factor of interest => Response in treatment => and control groups Other factors Other factors should be standardised and controlled: accuracy increases reliability increases variability decreases chance of systematic error smaller number of animals needed smaller General considerations Terrestrial mammals dry, clean and warm Adequate space without overcrowding Access to food and water Minimum cage sizes and stocking densities Cages from harmless material easy to clean, sterilize, escape proof Bedding to provide warmth and comfort, and to improve sanitation Environmental enrichment Barrier vs Conventional housing Incoming items are autoclaved, Restricted admittance??? air and water filtered, personnel Personal protective take shower and use sterile clothing protective clothing Quarantines Relative protection Arrival of food and bedding Breakdown means renovation Tap water of unit Isolators Filters in inlet and outlet of ventilation Animals handled through gloves Materials e.g. irradiated, autoclaved - inside through chemical disinfection Additional housing systems Cubical rooms Biohazard unit 1

2 Additional housing systems Scantainer Filter top cages IVC Laminar cabin Appendix A 2007 Physical space recommendations Environment Lighting Temperature Humidity Ventilation COMMISSION RECOMMENDATION of 18 June 2007 Noise on guidelines for the accommodation and care of animals used for experimental and other scientific purposes (2007/526/EC) Lighting 1. Light intensity Requirements vary between species High enough for working Rodents and rabbits nocturnal Albinos: retinal degeneration Same for all?? 2. Light s wavelength Does it matter? Rodents don t see red light? Most birds see UV-light (mating, foraging) Lighting 3. Light cycle Circadian-rhythm: metabolism, behaviour, hormones Research responses vary Breeding Change of light cycle => zeitgeber Standardised light cycle 12:12 Natural light - enrichment NHP, dogs, cats, farm animals Temperature Recommendations vary according to the species... Rodents Rabbits and dogs Pigs, goats Animals are able to adapt 1 o C changes...age, strain, physical state t new-borns, hairless, newly-operated Macro- and microtemperature Cage type, animal density, bedding Animals metabolism Temperature changes Metabolism varies The effect of temperature on the acute toxicity of drugs: 1. Klorpromatsine 2. Isoprenaline Responses vary 2

3 Relative humidity Recommendation 55 % ± 10 % % RH High RH Temperature more difficult to regulate Ammonia levels increase below 100 ppm Low RH Mucous membrane irritation Ring tail Energy consumption of rats in cold (+6 o C solid line) and in room temperature (+22 o C dotted line) with low protein (LP), medium protein (MP) and high protein (HP) diet Increased humidity or temperature can activate latent infections Ventilation Recommendation air changes per hour (other premises, or low stocking density 8-10 x) To bring fresh air To eliminate odors, gases, dust, excess heat and - humidity, infectious agents To avoid harmful draughts and noise Ventilation in a room vs. in a cage Dust and diurnal rhythm Acoustic environment (khz - db) Species differences in hearing ability Ultrasounds (>20 khz), infrasounds (<0,02 khz) Rats 0,25-76 khz, cats 0, khz, humans 0,02-20 khz Sensitivity to sounds varies: habituation, age, breeding status Hearing damage, hearing loss Albinos vs. pigmented Acoustic environment Startle - loud sudden noises Background noise Alarm system outside the sensitive hearing range of the animals Working techniques, equipments Results from handling Scand J Lab Anim Sci 32, , With holding by the scruff, response duration decreased from day one to day two and three - habituation 3

4 Results from handling Decapitation, necropsy, restrain, SC or tail vain injection caused no significant stress to witnessing rats (Sharp et al. 2002) Cage changes - heart rate habituate within 60 min Competent, confident and well-trainded handling Cage environment Cage type cage or pen solid vs grid bedding or not steel vs plastic size Other assessories enrichment nesting material nest boxes Cage environment Bedding Contact vs no-contact Hygiene: humidity, ammonia trasfer of soiled bedding increased aggression Chemical composition non-toxic wood species => metabolism Nesting material, enrichment transfer of familiar nest reduced aggression Harmless Cage environment Social factors Group housing Position in a group, stability, harmonious Pheromones Time of group forming Relationships with humans : dog, primates Animal density Recommendations (floor area: weight: number) Age (activity) Effects of environment and wounds on variation Growth (g) ,0 0,0 1,0 2,0 3,0 4,0 Spleen (mg) 600 EAT (mg) ,0 0,0 1,0 2,0 3,0 4,0 Control Nest Nest+box Nest+tube Environment Non-aggressive 100 Non-wounded 0 Wounded -1,0 0,0 1,0 2,0 3,0 4,0 Group number Adaptation From country to country From unit to unit From barrier to conventional Feed, bedding, light cycle, people, apparatus Before an experiment order animals on time make groups asap quarantine get acquainted Adaptation times 1-2 vk rodents 3-4 vk cats, dogs 5-8 vk primates 4

5 Housing of mice Solid bottom cages with bedding (once a week) Male mice are aggressive housing alone Bedding and nesting material nest boxes if insufficient nesting material Usually ad libitum feeding with pelleted diet (~3-5g/day) in hoppers Housing of mice Water in bottles (~6-7 ml/day), changed twice a week Sensitive to temperature changes and susceptible to water loss o C, 45-65%RH, low light levels (<60 lux for albinos) shelters, sensitive to ultrasounds Cage size for mice Cage size in NLAC 42 x 24,5 x 15cm (1029 cm 2 ) Decree 36/2006 Minimum floor area in stock 180 cm 2 30g mice ~10/cage (Apx A: 12 mice/cage) Minimum floor area in breeding 200 cm 2 Housing of rats Solid or grid floors, high enough Bedding and nesting material nest boxes if insufficient nesting material Cage changes twice a week Usually ad libitum feeding (~5 g/ 100g daily) in hoppers, cautios eaters Housing of rats Water in bottles (~10 ml/100g daily), changed twice a week Less sensitive to temperature changes than mice o C, 45-65%RH, <50-60 lux for albinos, 12-16h light optimal for breeding Cage size for rats Cage size in NLAC 48,5 x 28,5 x 20 cm (1382 cm 3 ) Decree 36/2006 Minimum floor area in stock 350 cm 2 <400g rats ~6/cage (Apx A: 3 rats/cage) Minimun floor area in breeding 800 cm 2 5

6 Housing of rabbits Solid or grid floors with resting area Roughage, hay blocks or chew sticks, area for withdrawal Raised area in cages (shelf), in pens structures that subdivide the space (shelf or something else) Pens for young females or does with litters, and when possible Cleaned weekly, high calcium content in urine => cleaning with acid Males urinate allover Housing of rabbits Restrected feeding with pelleted diet, high fiber (~5g / 100g daily), changes in diet gradually Water ~10 ml / 100 g daily o C, not less than 45%RH, adequate ventilation (ammonia level) Cage size for rabbits Cage size in NLAC 69 x 80 x 46 cm (5520 cm 2 ) Cage size for rabbits Decree 36/2006 Height Minimum floor area in stock <3kg 2000 cm2 30 cm >5 kg 3600 cm2 40 cm National Laboratory Animal Center 3 units rat and mouse barrier, rabbit barrier conventional animal area The quality and welfare of laboratory animals 2 R Education Consulting and informing regarding LAS Rederivation of strains Personnel Satu Mering (Head of department) Timo Nevalainen (Professor) Iiris Kasanen (Veterinarian) Katja Inhilä (Administrative coordinator) Heikki Pekonen (Research technician) Arja Konttinen (Supervisor of Snellmania and Bt1) Virve Immonen (Supervisor of Bt2) Rauni Tapaninen (Supervisor of Ca1 and Ca2) Päivi Hyvärinen (Supervisor of Sn barrier) 6

7 Laboratory animals Health, welfare and standarisation Mainly for the Kuopio university scientists Different health status in different units Sn and Ca bar: SPF animals, no entry Bt2: conventional animals, restricted entry Sn, Bt1 and Imaging unit: conventional animals Ca1: conventional animals and Mouse parvovirus Ca2: quarantine animals (MPV +?) General rules Try to cetralise your work in one department/unit Quarantines (48 h normal) General rules Doors are locked Application for the admission Alarm system Light cycle 7-19 y!!!! Do not put on the lights on the corridores!!!!! Ordering animals Animals bred by NLAC Mice: CD2F1, C57BL/6J Rats: Wistar Rabbits: WHHL From other vendors Order via NLAC Exchange of animals Contact NLAC (Iiris Kasanen and the head of the unit) Health monitoring reports Practical issues Delivery time (date, time) Receiving unit Who is paying How delivered (door to door or something else) Grouping, special cages, quarantines etc. Who to contact Quarantee Price list Animal price Per diem price Special services Delivery costs laskutus/ 50 rats of age 10 weeks for 3 months = 25,81 x 50 + (50 x 90 x 0,41) = 3.135,5 7

8 8 Inventory VALTAKUNNALLINEN KOE-ELÄINKESKUS ELÄININVENTAARI 2005 Inventaari tehdään tiistaisin ja perjantaisin Snellmania barrieri Tutkija ilmoittaa / kirjoittaa kyseisiä eläimiä koskevan koe-eläintoimikunnan antaman lupanumeron Kuukausi:Huone:Tilanneaik.Vk1Vk2Vk3Vk4Vk5Laji/Kanta:TiPeTiPeTiPeTiPeTiPeLaitos/Tutkija(Laskutus):Muutos ± Saldo Lupanumero:Muutos ± Saldo Lisä tietoja: NKKuukausi:Huone:Tilanneaik.Vk1Vk2Vk3Vk4Vk5Laji/Kanta:TiPeTiPeTiPeTiPeTiPeLaitos/Tutkija(Laskutus):Muutos ± Saldo Lupanumero:Muutos ± Saldo Lisä tietoja:nko:laatu/lomakkeet/inventaari2005snbar.docworking in laboratory animal unit Environmental conditions No pathogens Quarantine and adaptation periods No disturbances Transfer or housing of animals in other Transfer or housing of animals in other premises outside the Center Document guide Book keeping and instructions Working in laboratory animal unit Minor procedures in animal rooms Own materials and equipments (anaesthetics, needles, syringes) Euthanasia in autopsy room CO2 available CO2 available Carcasses into double plastic bags and into freezer Surgery/operation unit X-ray Waste management Protecting the health status Personal protective clothing Equipments and materials entering the unit Animals entering the animal area Health monitoring information Health monitoring information Animals leaving the animal area People entering the animal area The maintenance and ensuring of health standards in everyday practise liikenne.shtml Conclusion Researcher is responsible for the planning and practise of the experiment We are here to help Authoroties supervise Your behaviour may influence the y experiments of ohters DON T HESITATE TO ASK