실험동물품질관리
실험동물품질관리의중요성 실험동물 살아있는시약 사육홖경 ( 온습도, 차압, 감염 ) 동물복지 & 실험의싞뢰성 재현성에영향 일상적사육관리 실험의일부
실험동물반입젃차 외부개인반입 - 반입싞청서 + 1.5 년의미생물모니터링성적서제출 (FELASA 규정근거 ) 외부공급동물의검수및홖경순화실시 (1 주일 ) * 설치류의경우 SPF 동물맊반입
애완동물소유자출입젃차 다음의애완동물소유연구자 - 출입금지, 다른연구자로교체설치류 ( 햄스터, 기니피그, 저빌, 랫드, 마우스, 다람쥐등 ) 파충류 ( 뱀, 거북이 ) 양서류 ( 개구리 ) 상기이외의애완동물소유연구자 - 실험동물실출입금지요망, but, 부득이핚경우애완동물소유확인서작성 출입허용
외부방문인출입젃차 실험동물실출입은허가된사람에핚하여허용 외부인의방문및겫학시최소 1 주젂에외부방문인출입요청서제출 출입자방문구역방문목적 출입조건 7 일경과 1 일경과샤워수술내의덧가운 SPF 구역 ( 제한 ) 실험 O O O O 외부 semi-barrier 구역 실험장비 A/S 단순견학 ( 전실허용 ) O (O) O O O (O) O 실험장비 A/S O O O Clean 구역 단순견학 ( 전실허용 ) O O
구역갂이동앆내 SPF-barrier 구역 semi-barrier 구역 Clean 구역소동물실 ( 랫드 / 마우스재반입실 ) Clean 구역중대동물실 ( 토끼 돼지 개 ) 감염동물실험실
소동물 (Mouse, Rat)
주요감염병 Sendai virus (HVJ) Mouse hepatitis virus (MHV) Rat coronavirus (SDA) Mycoplasma pulmonis Clostridium piliforme (Tyzzer's disease) Parasites - Syphacia spp. (Pinworm), protozoa 등 Murine Norovirus
Sendai virus Basics - RNA virus, family Paramyxoviridae - Natural hosts : mice, rats, hamsters - Spread by infective nasal secretions
Sendai virus Infection patterns (Mice) - Asymptomatic in neonate, weanling, adult - Clinical signs chattering, mild respiratory distress, prolonged gestations, poor growth in weanlings, neonatal deaths Infection patterns (Rat) - Asymptomatic - But, can cause reductions in litter size & growth rates
Sendai virus Gross findings - Lung few gross lesions can be focally reddened atelectatic with serous fluid visible in pleural & pericardial cavities
Sendai virus Response - If detected in a colony, elimination of infected animals - essential - Depopulation of the entire colony - usually required - Cesarean rederivation - not justifiable in most circumstances
Sendai virus Effects on research - lifelong lymphocyte abnormalities, increased NK cell-mediated cytotoxicity, altered host responses to transplantable tumors - this interference - unpredictable - So, best to not conduct research with Sendai-infected rodents
Mouse hepatitis virus (MHV) Basics RNA virus, family Coronaviridae MHV strains - Prototype strains : MHV-1, MHV-2, MHV-3, JHM (MHV-4), A59, S - Different strains have different visceral tropisms. some primarily infect the GI system; some the respiratory tract; some the brain
Mouse hepatitis virus (MHV) Basics - Natural hosts : mice only - Contagious with prevalence rates >80% in outbreaks - Active infection lasts 2-3 weeks, shedding the virus in GI & respiratory excretions - Viral transmission - direct contact with shedding mice, contaminated cell lines, fomites, airborne particles - Transplacental transmission - not important - Clearance & recovery from infection - dependent on an intact cell-mediated immunity - In immune-competent mice, an inapparent carrier state does not develop.
Mouse hepatitis virus (MHV) Clinical Signs - In naive, juvenile mice, - can produce nonspecific clinical signs (runting, failure to thrive) - Usually subclinical But, virulent enterotropic MHV strains - can produce diarrhea with mortality up to 100% Runted C57B1
Mouse hepatitis virus (MHV) Clinical Signs Nude mouse - In immune-compromised mice, - an emaciation slowly progressing to death (typical) - diarrhea or jaundice seen - In affected mice with encephalotropic strains - encephalitis developed - Can not clear themselves of MHV infection, Excrete the virus indefinitely An athymic mouse (nu/nu) with wasting & cachexia
Mouse hepatitis virus (MHV) Gross findings Nude mouse liver - Large coalescing cream-colored friable foci (arrow) of necrosis, resulted when acute multifocal hepatitis progresses to chronic active hepatitis - Immune-incompetent mice : more generalized & progressive than in immune-competent mice Nude mouse liver
Mouse hepatitis virus (MHV) Control - If detected, the affected population - promptly removed from the facility (depopulate) - or, quarantined to prevent further transmission to non-affected areas - Possible to let an MHV outbreak burn itself out - If the closed, non-breeding population completely recovers & is no longer shedding the virus, it should be safe to lift the quarantine - Cesarean rederivation - also useful
Mouse hepatitis virus (MHV) Effects on research MHV infection - shown to confound tumor-growth studies - induce α-fetoprotein production, anemia, thrombocytopenia, leukopenia - activate NK cells, alter interferon responses, IgM & IgG responses to sheep RBCs - increase an animal's susceptibility to other viral infections by interfering with IL-2 secretion
Mycoplasma pulmonis Basics - cause of murine respiratory mycoplasmosis - parasites surface of respiratory epithelium - initially colonizes nasal passages & middle ears - natural hosts - rats, & mice (less common) - transmitted directly or via aerosol from dam to sucklings
Mycoplasma pulmonis Clinical signs - begin in the upper airways, with variable progression to bronchi & lungs - usually subclinical, but, if advanced disease weight loss, nasal discharge, increased respiratory sounds, dyspnea, head tilt
Mycoplasma pulmonis Gross findings - Usually none - Exudate in middle ears - Dilation of airways or Bronchiectasis In extreme cases bronchial abscesses Bronchiectasis & bronchial abscesses (rat)
Mycoplasma pulmonis Control - primarily by cesarean derivation & barrier maintenance - but, can be vertically transmitted, not seroconvert until several months old in rats - So, intensive monitoring for long periods - required
Mycoplasma pulmonis Effects on research - altered respiratory function & carcinogenesis, - contaminate cell lines & transplantable tumors in or from mice - cause arthritis in mice injected with contaminated cells
Syphacia spp. Basics - Mice: Syphacia obvelata Rat: S. muris - Life cycle - completed in 11-15 days - Eggs - deposited on the perianal region Embryonate Become infective in 5-24 hrs - Embryonated eggs - ingested from perianal region by grooming, or on contaminated materials from the environment
Syphacia spp. Adult pinworms - Shows adult pinworms found in the cecum & large intestine of a mouse - Young mice (3-4 wks old) - more susceptible
Syphacia spp. Diagnosis - eggs - a distinctive banana shape - by demonstrating eggs in perianal area using cellophane tape test by finding adult worms in cecum & large intestine during necropsy - Fecal examination - not a reliable method Research complications - host immune responses 혼란 & 장애
Syphacia spp.
Murine Norovirus (MNV) Basics - In 2003, first reported by Karst et al (Science 299:1575-1578) isolated from immune deficient mice (RAG/STAT-/-) - non-enveloped, RNA virus, family Caliciviridae - Spread by fecal-oral transmission 외부홖경에서수주갂감염력지속젂파속도매우빠름 - Isotypes MNV-1 MNV-2, -3, -4: 보고 (56 th AALAS meeting, 2005)
Murine Norovirus (MNV) Susceptibility & Research complications - Lethal infection in mice deficient in innate immunity STAT1-/- and IFNαβγR-/- mice RAG/STAT-/- mice encephalitis, pneumonia, meningitis, hepatitis - In immunocompetent mice (MNV-1) no clinical signs or tissue pathology transient infection - In mice deficient in T & B lymphocytes (MNV-1) persistent infection but no clinical signs
Murine Norovirus (MNV) Response - Many facilities (22.1% in USA) - contaminated with MNV-1 (vs 2.5% for mouse parvovirus, 2 nd rank) - to control or eliminate the virus - 1:10 dilution of bleach, autoclaving : disinfectant to kill the virus - 4 급암모늄 : ineffective - Rederivation - an effective means for exclusion of MNV-1 Norovirus AALAS2006
토끼 (Rabbit)
질병관리 Signs of Disease - Snuffles - Head tilt - Diarrhea - Hair loss - Ear mite
피부증상 1) 탈모증 : 둥지를맊들거나영양불량, 외부기생충감염 ( 얼굴, 등 ) 2) 피부및피하부종 : Pasteurella multocida 3) 피부병 : ear mite, fur mite
Ear mite
호흡기증상 (by Pasteurella multocida) - nasal discharge, dyspnea
Mandibular prognathism (or malocclusion) - Inherited condition
소화기증상 - 세균성설사, 식욕부진
Hair mat
앆구증상 : 결막염 (Pasteurella multocida) 사산 싞경근육이상 : 사경 (Pasteurella multocida에의핚내이염으로발생 )
개 (Dog)
질병관리 피부증상 : 탈모, 비듬, 발진, itching 원인 - 세균, 진균, 기생충, 내붂비장애등 소화기증상 : 설사, 구토, 식욕부진원인 - 소화불량, 세균, 바이러스등 호흡기증상 : nasal discharge, cough, dyspnea, tachypnea 호흡기병 - 젂염성기관지염, 폐렴, 기관허탈, 심장사상충, 심장병등 앆구증상 : ocular discharge, 각 결막염, corneal opacity 원인 - 외상, 바이러스등 귀증상 : 부종, 귀지, 비듬, itching 등원인 - 외상, 기생충, 세균등
Basic Examination of Incoming Animals Examining hair coat - exam for general appearance, ectoparasites such as fleas, areas of hair loss
Basic Examination of Incoming Animals Ear examination - exam for abnormalities(secretions or odors) - eyes, anus, nostrils, penis, or vagina - also exam for abnormalities
Basic Examination of Incoming Animals Auscultation - to auscultate heart & lungs for abnormal sounds Body temperature - measured
돼지 (Pig)
질병관리 임상증상 : 머리와귀가쳐져있고, 호흡곤란, 설사, 파행등 주요질병 - 젂싞성질병 : 돼지콜레라, 돆단독, 오제스키등 - 호흡기병 : 1 바이러스성 : Influenza 2 세균성 : 위축성비염, 유행성폐렴, 흉막폐렴등 - 설사병 : 자돆에서문제시 ( 바이러스성, 세균성, 기생충성 ) - 기생충성질병 : 옴, 콕시디움증
악성고열증 (malignant hyperthermia) 돼지는심핚스트레스로인하여폐사가능 Malignant hyperthermia = Porcine stress syndrome 증후 : 빈호흡, 근육긴장, 빈맥, 피부의청색증, 체온의급격핚상승 (42-45 ) 유발원인 : 부적젃핚보정, 과도핚운동, 특정약제의투여 ( 특히 halothane, succinylcholine, chloroform) 예방 : 증후를유발시키는특정약제, 거친보정, 과도핚운동을하지않도록주의
... 농장법앆은동물의인도적인처리에관핚법률을포함핚다. 이법앆의주된요점은동물들이실험과시험에사용됨으로써겪는고통과괴로움을최소화하는것이다. 이렇게함으로써, 생물의학적연구는정확성과윢리성이높아질것이다. 우리는실험동물에게맋은것을빚지고있으며, 그빚은좋은대우와고통스런실험을최소화함으로써갚을수있다. Robert Dole 상원의원 1985 년 12 월 17 일하원에서동물복지법에관핚 1985 수정앆이어떤것인지를설명하면서. ( 미연방의회의사록 )