----start--- lab animal medicine 10/27/98 quick intro: this is a core course as it has been for 11 years. blame dr rozmiarek for it. how many of you have an interest in practicing lab animal medicine in some way? ASLAP did a survey 7 yrs ago of OSU grads 5 years out. about 50% of them had at one point officially functioned as a lab animal medicine specialist. so you should know some of this stuff. typically, vets in practice need to know some of these things. handouts: we have many guest instructors. they have a lot of handouts. collect them and keep them. we have 11 classes and we'll probably get 11 30 page handouts. exam 1: first 6 lectures. take home exam, one week to do it. use recommended reading sources. exam 2: 1 hr, closed book. covers things you need to remember and understand. there will be a 2 hr review after last lecture, before final. probably this is the major study session you will need. please fill out the instructor review form and bring it to the final exam. btw the takehome midterm is to be a learning experience and may perhaps be resubmitted after initial grading... course grade: takehome exam = 20%; final is 80% guest lecturers are supposed to be done at 10 til the hour and you should tell them if they are going over their time. "Current and future policies..." handout - read that. it's important to remember legal responsibilities of veterinarians - a licensed vet must be responsible for animal care wherever animals are used, as per AWA which is US law. only one federal law, the Animal Welfare Act (AWA) which is enforced by USDA, protects lab animals here - meaning, warm blooded vertebrates. Now, the USDA wrote regulations defining lab animals as excluding domestic mice and rats - so they are not covered. also excluded are farm animals used in production agriculture. a cow on a project investigating milk production is not covered. if that cow is on a heart surgery study, it is covered. also every state has an animal anti-cruelty law, and many cities have them. NY state has the state dept of agriculture overseeing lab animal use. so, vets serve as inspectors. all places using lab animals - zoos, circuses, schools, pharm companies - all register and get license to use animals and are inspected by USDA at least 1 x per year. but not places using only mice, not dairy farms, etc. community regulation: cambridge, mass has their own lab animal inspector - tht person looks at animal use there. laws are enforced. USDA inspector comes unannounced, and if there are violations of policy, they issue a citation - and you go to court. you have the right to appeal, etc. that takes time. if an animal is in dire need of help, USDA can intervene with local authorities. permanent fix takes time. may fine institution, put someone in jail, whatever. USDA cited one primate research lab here some time ago - in court, was found not guilty enough about law. that's the only law we have. oh, one other law about shipment of animals - 28 hr law. animals beinng shipped have to have an opportunity to leave conveyance, stand, walk, stretch, before being reloaded 1 time in each 28 hr period. policies: not laws. you need not comply unless you want the benefit. PHS: public health service policy - PHS policy - this defines animals as any warm blooded vertebrates - no exclusions. there are about 12 principles of good animal care and use that are part of this policy. this policy pertains to any animals used in ways that get federal funding. if you do not want federal funding, you do not have to comply. so it is most widespread in academic institutions.if anyone here at Penn wants grant money, the whole school must comply. any institution wanting federal funds must provide an "assurance" describing your animal use program in detail to the PHS. PHS reviews it. if there are problems, they negotiate with institution. then they eventually give a PHS assurance number - you have to have that number to apply for federal funds. after USDA citation here, PHS found Penn deficient and gave a year to comply - penn didn't comply. for the next year - 1984 - Penn didn't qualify for federal funds. up til then, Penn was getting 90 million/year in grants. Penn was in compliance by the next year. so, sometimes policy is more effective than law, in short term. we now get about 300 million/yr in grant money... but back in 1986 after the moratorium, penn hired dr R as lab animal vet to organize things, to get into compliance, and to create good animal care and use programs. they did several things: established a lab animal medicine residency program, decided to strive to meet AAALAC accreditation standards which are higher than PHS standards - wanted accreditation for med school, vet school, and campus. so far med school is, and vet school is in progress. there are other things - FDA and EPA have "good laboratory practices" - GLP. these groups oversee food additives, medications, drugs, environmental sprays, cleaners, etc. the GLPs include animal care and use standards. they follow AAALAC type of rules. so the pharmaceutical companies are generally covered by these GLPs, which means their mice and rats are protected. both the PHS and USDA require you to have a committe - the IACUC. each USDA registered or PHS assured facility must have an IACUC of at least 5 people, including at least one veterinarian with experience and training in lab animal medicine. large places have ACLAM certified vets; small places often have local vet consultants. that vet has legal responsibilities. guidelines: Guide for the Care and Use of Laboratory Animals - this is a must have for those consulting vets Guide for the Care and Use of Agricultural Animals - being revised - covers the animals used for food and fiber - which are included under the AAALAC rules. AVMA panel on euthanasia - last guidelines published in 1993 OSHA publications - guidelines and requirements for institutions there are many more, too... What goes on at Penn? we identify a responsible individul - the vice provost for research. this person has aadministrative responsibility to ensure that we comply with stuff. he does this through various other people. the IACUC: we have two subcommittees - between four and five thousand research protocols are going on here. each one needs IACUC approval. they have to fill out forms explaining the protocols. the IACUC gets form, reviews it carefully...they approve about 5% off the top - then about 94% require changes/clarification prior to approval. it goes back and forth til it is right and then it is approved. the IACUC visits each animal room on campus at least 2x yearly. they spend 28 days visiting these facilities to ensure they comply with the guide. they also review animal care and use program every 6 mos. IACUC: -reviews protocols -reviews animal care prograams ad policies -visits housing -visits labs -responds to concerns and inquiries university veterinarian - dr r - responsible to ensure animals are appropriately cared for. ULAR - lab animal resources group. provides proper husbandry. the IACUC sees that ULAR is doing job correctly via reviews every 6 mos. one other thing - we also have academic section of lab animal medicine - that's who teaches this class and the elective. they put on seminars, too. organizations: ALAAS ACLAM ASLAP LAMA Association for gnotobiology lab animal veterinarians: the only ones with ethical and legal responsibility here. we have to be animal advocates. final poinnt - always contact an expert if you need help with a lab animal issue. ---break--- dr silverman from hahnemann/allegheny/whatever the hell they are calling themselves mice and rats: what you should know :) he's going to follow the handout... rats and mice are the most widely used animals in research. about 1.4 million animals were used last year.this doesn't count rodents - they do not get counted b/c they are excluded from the AWA. about 24 million rodents are used. we eat about 8 billion animals a year, though. many people say rodents do not count. however, they are fully sentient animals. maybe they are not your favorite pets. but they are animals. you need to understand that and treat them appropriately. mice originated in northern africa/asia/eastern europe and migrated to the new world (ships). same with rats - from asia, eastern europe. rattus norvegicus - may or may not be from norway :) there are many types of rats/mice he's showing some slides of different kinds of rats. rattus rattus is the wild typical rat lab rat is rattus norvegicus - hooded rats often kept as pets many kinds of mice - lab mouse is mus musculus - house mouse. nude mouse - the hairless, athymic mice used for immunological research. these are mus musculus with a single gene mutation. inbred vs outbred/random bred inbred == strains of mice. all are mus musculus but different strains: these have a / in the name: BALB/c, DBA/2, etc. these mice are homozygous at 98.6% of any loci. inbreeding = at least 20 generations of brother x sister matings like dr silverman's neighbors. you end up with populations of essentially clones. breeding two inbred strains = hybrid inbred x inbred = hybrid LEW/Ma x BN/Mai --> LFNF1 outbreds: any animals not inbred. these have a : in the name Blu:LE = Long Evans rat Tac:N (SHR) = spontaneously hypertensive rat outbreds are also called "stocks" transgenic animals: if you take a gene or DNA from one species, and put it inot another one (mouse), you have a transgenic mouse. or, if you remove a gene, you have a knockout animal. importance of transgenics: by putting a gene onto a normal background you can see the effect of that gene. and that animal can reproduce similar animals. or if you can remove a gene, you see the effect that you remove. this can help you identify the effect of the gene. Husbandry: these mice on the slide are in little plastic shoebox things in racks. in the cage is bedding. is it better to have a mesh floor or solid floor? bottom line - solid floors are better - helps to decrease podal dermatitis. solid floors require bedding. primary functions of bedding: absorption of urine, fecal water, drips from water bottle, etc; and comfort - nesting, burrowing, having soft surfce to walk on. common types: hardwood chips, paper shreds, corn cob. however, for those who work with pocket pets, most kids use pine shavings or cedar shavings - esp cedar to reduce odor. this is a problem. these aromatic oils induce liver enzyme production which causes clinical effects such as short pentobarbitol sleep time. the other point - two most common cage types: polycarbonate plastic - most common - with wire mesh top for food and water stainless steel - too heavy, expensive the plastic cages can also have special filter tops - microisolator cages. the microisolator name is trademarked but anyway...it's a big petri dish to keep animals in :) these cages keep something in, or keep something in. key to successful use is to change the cages under a laminar flow hood. you cna't just open it in the middle of the room. typical housing requirement - temp 68-72 F; if comfy for you, comfy for them. temp up to 80 for nudes. humidity 30-70% (relative) food/water info in handout btw, mice/rats are coprophagic and get some vitamins that way. the virtual rodent tour: teeth: incisors, no canines or premolars, then molars. 1/1, 0/0, 0/0, 3/3. space b/w incisors and molars == diastema the incisors grow throughout life. they have to occlude correctly to avoid overgrowth of the incisors.otherwise they overgrow and curl back into the jaw, or up through hard palate into brain, or up the front of the face. if the mouse/rat has malocclusion, you have to trim the teeth every 2 to 4 weeks. you can teach clients to do this. eyes: because many are albino, you have to keep in mind the light levels. at 50 footcandles, which is about what's in here now, all the rats got retinal lesions. at 30 footcandles (dimly lit bar) lesions start to decrease. current standard is 20-30 footcandles (300 lux) harderian gland - sits sort of surrounding the eye and secretes porphyrin pigments - sometimes people see hypersecretion and they think it is red tears, but it isn't. you cann use UV light to see it fluoresce. hypersecretion is caused by any nonspecific stress - shipping, heat, broken leg, whatever. with hypersecretion you see chromodacryorrhea (which means excess pigment being produced by Harderian gland...). sometimes secretions go down nasolacrimal duct to nares. remember cause is nonspecific stress siallodacryoadenitis virus: SDAV - a coronavirus, most common virus of rats. very high morbidity, almost no mortality. blows over in a few weeks. spp specific. SDAV is a disease of rats only. clinically, you see increased harderian gland secretion, swollen LNs and salivary glands, and sneezing. you hear a whole room sneezing. other dzs cause sneezing but first thing to think of is SDAV esp when whole room doing it. some animals show only a few signs.others show all the signs. sometimes with SDAV you get an ocular ulcer. how? the Harderian gland duct gets clogged with necrotic debris. squamous metaplasia occurs. eyes dry out. ulcers occur. dx: serology - ELISA - best way. a bit of blood, send it in, that's it. effect on research: animals feel sick, can exacerbate other dzs, complicate respiratory and ocular studies as well as carcinogenesis studies, reduces IL1 production. to eliminte it - quarantine it and wait 4-6 weeks. let them all get it and recover. other ways: rederivation, depopulation. zymbal's gland: in rats, mice, insectivores - but only clinically relevant to rat. it's near the external ear canal, hidden by lacrimal gland and parotid. can get tumors of the zymbal's gland. can be huge. what kind of sadist would let a tumor get big enough to take that picture? dunno. rat and mouse have single lobed left lung and 3 or 4 lobes to right lung. for intracardiac stick, go right or left, not important. going right will not help you miss lung like some say. mycoplasmosis: mycoplasma pulmonis aka chronic respiratory dz: in mice and rats, causes bronchopneumonia which is often clinicallly silent until animal is stressed. signs typical of any bronchopneumonia - dyspnea, inappetance, etc.can cause rhinitis, too. can get into tympanic bulla and cause head tilt, or uterus and cause abortion or infertility. no real tx for this either. baytril, maybe. nothing really works. you can try to control it but it tends to come back. hunched up, in corner, fur ruffled - typical sick mouse. major effects: increased resp tract tumors, as well as screwing up all research studies since animal is very weaak, and sick. may kill animal but not always. dx: ELISA or culture Sendai virus - common paramyxovirus in lab animals and pocket pets. in rats/mice but seen more in mice. main points: usually no signs of disease. but can become clinical esp in mice - causes interstitial pneumonia. dx: serology - ELISA. effects on research: immunosuppression. lymphocyte abnormalities, icreased NK cell mediated stuff...all research is affected how do you get rid of it? quarantine 4-6 weeks - it will burn out. hantavirus: carried by wild rodents. a disease of people no clinical signs in rodents not isolated in labs (yet) for pocket pets, in some areas, this may be a concern re zoonosis skipping helicobacter two factoids: 1. rat/mouse have cardiac muscle in pulmonary veins. not sure why. 2. sometimes you necropsy a mouse and see calcification around the heart - in BALB/c, some other strains. no clinical relevance to this. stomach: divided into two parts - forestomach/aglandular and glandular corpus. physical ridge - limiting ridge - separates them. what's forestomach for? not sure. removing it does not do much. may aid CHO digestion. who knows. also - esophagus of rat/mouse has cornified stratified squamous epi, normally. abdominal stuff: mice have gall bladders. rats do not. liver: mouse hepatitis virus: MHV - significant in mice only - many forms - neuro, resp, enteric (most common). a coronavirus, fairly spp specific, generally not causing problems. in nude mice though, is a problem - causes severe necrotizing hepatitis. but in immunocompetent mice only rarely gets to liver to cause hepatitis, usually doens't do anything. doesn't bother animal. effects on research: it's immunosuppressive. to eliminate it: quarantine and burnout 4-6 wks syphacia obvelata (mouse), s.muris (rat) - common parasite in lab and pocket pet rodents. usually no signs. these are pinworms. usually clinically silent. dx via fecal flotation, necropsy, or scotch tape prep - put tape near anus, then look under scope. eggs get picked up on tape. tx: ivermectin is in the books, but doesn't work. fenbendazole works - one week on, one week off, mixed into food. it will work. also that has ovicidal activity too. urinary system - it's easy to catheterize females but almost impossible to do a male b/c cath goes up into the bulbourethral gland diverticulum and doesn't make the sharp turn into the bladder.don't even bother trying. dz of rats/mice esp rats - chronic nephropathy/nephrosis/renal dz - of aging rats (and mice) - over a year, up to 2 yrs...kidneys are large, pitted, pockmarked...animal starts going downhill - dx as in dog/cat, tx - try lowering protein in diet maybe but (oh boy) hard to do that. no good diet. (mouse rat life span maximum 2-3 yrs)(one rat lived 4 yrs) skin: ringtail - rats, sometimes mice. you see annular constrictions around the tail of the animal. in immature animals. it is an avascular necrosis - idiopathic. associated with low humidity. no tx. tip of tail falls off, that's it. barbering - overgrooming - a psychological condition of mice, rarely rats. one mouse is the barber and he bites off fur from other ones. looks like it was shaved. take barber out of cage. fur mites: myobia musculi, myocoptes musculinus - they are fur mites, not burrowing mites. they are on the fur. fairly common in pets and labs. mice scratch, get excoriations. tx: take some ivermectin 1% cattle stuff, take 5 microliters and put it on the mouse's neck q 10 days x 5 times. mite wounds can mimic fighting wounds. bite wounds from fighting are often near tailbase - separate mice. but sometimes hard to tell. mammary tumors - mice and rats. mammary tissue is around most of the trunk, except head/limbs/tail.you caan get mammary tumors in weird places - just behind elbow or something. tumors may be benign fibroadenoma - remove surgically, animal does fine (rat) or in mice, usually adenocarcinoma that is infiltrative and metastatic to lungs, and hard to remove. no tx in mice. ---end---