Email: evan@sun5.nlac.gov.tw 1. 2. 3. 1. 2. 3. 4. 5. 1. (1) (2) 2. 3. 1. (1) (2) (3) 2. (1) (2) (3) 1
1. 2. (1) isolator germfree gnotobiotic (2) barrier SPF (3) conventional (4) 3. (1) (2) - IACUC Institutional Animal Care and Use Committee 1. 2. 3Rs 1. 2. SHR AKR nude T beige xid B scid B T op 1995a 2
mouse rat substrain BALB/cByJ short chain acyl-coa dehydrogenase (SCAD) BALB/cByJ BALB/cJ BALB/cByJ BALB/cJ Jackson 4 BALB/cJ BALB/c 1993a phenotype assessment Wood, 2000 / 1. residual heterozygote substrain 2. coisogenic strain 3. genetic contamination 1. gene frequencies 2. 3. inbreeding 1. 2. genetic profile 3. 1. -- loci genotype 2. genetic profile 3. 3
1993 2000 genetic markers 1. 2. isozymes 3. H-2 halpotype mixed lymphocyte culture 4. skin grafting 5. C-band, T6 markers 6. genomic DNA 7. breeding performance 2001 barrier specific pathogen free SPF SPF lactic dehydrogenase elevating virus, LDHV Klebsiella pneumoniae B Proteus mirabilis 1 interleukin-1, IL-1 cytotoxic activity Pseudomonas aeruginosa A IL-1 1. Waggie et al., 1994 2. 3. 4. 5. 6. 1993c 4
82 1993b 1995a, b, 1996, 1997a, b germfree, GF gnotobiotic, GB 2001 GF/GB GF/GB Falk et al., 1998 CIEA human c-ha-ras gene CB6F1-Tg Hras2 dimethylhydrazine dihydrochloride DHM 100 DHM GF 100 SPF 0% Clostridia GB DHM 10 Clostridia AC stock GB SPF Narusbima et al., 1998 Horie et al., 1999a, b Ohno et al., 2001, 2001, 1996 1. 2., 2001, 1996 5
3. 29.4 4.4 18 26 20~24 30~70% 45~65% 4. / 10 15 12~18 25 ppm 5. albino rat 130 325 lux 1 325 400 lux 40 lux hibernation 17 18 6. 85 7., 2001, 1996 8. 18 6 21 C 3 C 9. ph2.8~3.2 5ppm C 10. 11. 6
Whitten Bruce 12. 13. 1 A. -C57 -DBA/2 B. scid C. AKR 2 3 1995 1996 2001 ring tail SPF 82 1993b germfree, GF mucin SPF, conventional SPF CIEA 2001 7
1993b refinement reduction replacement animal welfare 15 1. (1). (2). 2. 3. 4. 5. 1 2 6. microisolator cages 7. 2000 8. Mouse Antibody Production (MAP) Test Rat Antibody Production Test (RAP) test 8
BALB/cByJ C57BL/6J \ BALB/cByJ C57BL/6J (1) (2) a. 67% 22% ( ) ( ) NO TNFα CD4/CD8 b. IL-1 (3) A ---- (DBA/2J ) N NIH non-inbred stock 4 SDN SD 1% 20 C57BL/6 DBA/2 inbreed strain BALB/c B6CF1 x F1 hybrid CB6F1 AKxD recombinant inbred F1 B6NxAKN 20 A-+/nu coisogenic inbred mutant gene A-nu/nu BALB/cByJ-Hfh11 nu congenic inbred Cby.Cg-Foxn1 nu 10 nu Hfh11 nu Foxn1 nu / BN/SsN F344 Lew/SsN SHR WKY Alp1 B B B A B Amy-1 B A A A A Amy-2 A A A A A Es-1 A A A A A Es-2 C A D A D GC A A A A A -- Hbb A A B A A Adh-2 B C C B B Es-10 B A B A B Acon-1 A B B B B Akp-1 A A A A B Es-4 B B B A B Fh A B A B B Pep-3 B B A A B 9
Pneumonia virus of mice * 1 * S Reovirus type 3 * - S Sendai virus * * S Lymphocytic choriomeningitis virus * * S Theiler s encephalomyelitis virus(gd ) * * S Minute virus of mice * - S Mouse hepatitis virus * - S Mouse adenovirus * - S Ectromelia virus * - S Kilham s rat virus - * S Sialodacryoadenitis virus/rat corona virus - * S Hantaan virus * * S Mycoplasma pulmonis * * * S Clostridium piliformis Tyzzer s disease * * * S Bordetella bronchiseptica * * * C Corynebacterium kutscheri * * * C Salmonella spp. * * * C Pasteurella pneumotropica C Pseudomonas aeruginosa * C Staphylococcus aureus C Myobia spp * * D Aspiculars tetraptera * * M Syphacia obvelata/muris * * M/Ct Hymenolepis spp * * M Spironucleus spp M Trichomonas spp M Pnenmocystis carninii - - * H 1. * - 2. ( ) (species) 3. C-culture, Ct-Cellophane tape, D-Dissecting microscopy, H-Histopathology, M-MIF direct smear method, S-serology 4. Clostridium piliformis Mycoplasma pulmonis 5. 89 2 18 2 3 10
35 ring tail 22% 22 25 45 50% water deprivation scurvy C 15 20 mg/kg 100 audiogenic seizures 85 35 DBA/2J 100 chromodacryorrhera photoreceptor 130 325 lux degeneration ulcerative pododermatitis 25 250 ppm murine respiratory 4 6 mycoplasmosis 25 ppm foreign body pneumonia 11
1. 1993;1(1): 7-9. 2. 2001 3. 2000 4. 2000 5. 2001 6. 1995 7. 1996 8. 2001 9. 1996 10. SOP 600-617 1993a 11. SOP 700-715 1993b 12. 1993c;1 (3):9-10. 13. I 1995a;2(4):25-31. 14. II 1995b;3(1):15-19. 15. III 1996;3(3):15-21. 16. IV 1997a;4(3):18-21. 17. V 1997b;5(2):2-12. 18. 2001 19. Falk PG, Hooper LV, Midtvedt T, Gordon JI. Creating and maintaining the gastrointestinal ecosystem: what we know and need to know from gnotobiology. Microbiol Mol Biol Rev 1998;62:1157-70. 20. Horie H, Kanazawa K, Okada M, Narushima S Itoh K, Terada A. Effects of intestinal bacteria on the development of colonic neoplasm: an experimental study. Eur J Cancer Prev 1999a;8:237-45. 21. Horie H, Kanazawa K, Kobayashi E, Okada M, Fujimura A, Yamagiwa S, Abo T. Effects of intestinal bacteria on the development of colonic neoplasm II. Changes in the immunologic environment. Eur J Cancer Prev 1999b;8:533-7. 22. Narushima S, Itoh K, Mitsuoka T, Nakayama H, Itoh T, Hioki K, Nomura T. Effect of mouse intestinal bacteria on incidence of colorectal tumors induced by 1, 2-dimethylhydrazine injection in gontobiotic transgenic mice harboring human prototype c-ha-ras genes. Exp Anim 1998;47:111-7. 23. Ohno K, Narushima S, Takeuchi S, Itoh K, Itoh T, Hioki K, Nomura T. Effect of bacterial metabolism in the intestine on colorectal tumors induced by 1,2-dimethylhydrazine in transgenic mice harboring human prototype c-ha-ras genes. J Exp Clin Cancer Res 2001;20:51-6. 24. Waggie K, Kagiyama N, Allen AM, Nomura T. Manual of microbiological monitoring of laboratory animals. 1994 NIH publication No. 94-2498. 25. Wood PA. Phenotype assessment: are you missing something? Comp Med 2000;50:12-5. 12