Key words: Escherichia coli O157:H7, diarrhea, animal model, infant rabbit. probiotics

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1 Key words: Escherichia coli O157:H7, diarrhea, animal model, infant rabbit. probiotics

2 Table 1 The virulence factor genes were detected by polymerase chain reaction (PCR)

3 Fig. 1 The group of symbol ~was challenged with EHEC O157:H7 strain 275 only. The group of symbol ž was challenged with EHEC O157: H7 strain 275 after treated S. faecalis T-110, C. butyricum TO-A and B. mesentericus TO-A. Fig. 2 The group of symbol ~was challenged with EHEC O157:H7 strain 275 only. The group of symbol ž was challenged with EHEC O157: -17 strain 275 after treated S H. faecalis T-110, C. butyricum TO-A and B. mesentericus TO-A. Fig. 3 The group of symbol ~was challenged with EHEC O157:H7 strain 275 only. The group of symbol ž was challenged with EHEC O157: H7 strain 275 after treated S. faecalis T-110, C. butyricum TO-A and B. mesentericus TO-A.

4 O157感 染 ウサ ギ を用 いた 生菌 剤 の評 価 Fig. 4 ment Table 2 The of photograph the The presence number/slice(-: shows microvilli attached H7 strain 275 and efface- ( ~20,600) of attaching 0, +: *: Rabbits were inoculated EHEC O157: 1303 and effacing(ae)lesion <<10, ++: in the intestine. AE lesion 10<) with EHEC O157: H7 strain 275 after treated with S. faecalis T-110, C. butyricum TO-A and B. mesentericus TO-A 考 い る.こ の よ う な 作 用 か らEHECの 察 腸 管 感 染 症 に対 して は 一 般 的 に抗 生 剤 が 使 用 さ と 見 な さ れ て い る 牛 に お い て もEHECを れ るが,抗 生 剤 投 与 に よ って 耐 性 菌 が 出 現 す る可 る手 段 の1つ 能 性 が あ る こ とや,O157感 検 討 さ れ て い る7). 染 に対 して は あ る種 の 抗 生 剤 を使 用 す る こ とで 殺 菌 時 に 菌体 内 に貯 留 し て い るVTが 放 出 さ れ,HUSの 引 き金 に な る と こ れ ま でEHECの 感 染実 験 モ デル として はマ ウ ス8)9),ブ タ10)11),ウ サ ギ4)5-,鶏 ヒ ナ12)13)な ど が 報 告 さ れ て い る が,マ る.一 方,生 ヒ ナ は そ れ ぞ れ 取 り扱 い,あ ウ ス は 下 痢 を 発 症 せ ず,ブ 的 に排 除 し,腸 内細 菌 叢 を正 常 化 す る作 用 か ら, 難 し い な ど の 問 題 が あ っ た.ま 腸 管 感 染 症 の 治 療 や 感 染 予 防 の効 果 が 期 待 さ れ て の 感 染 モ デ ル は 無 菌 動 物,あ 平 成10年12月20日 排 除 す と して プ ロバ イ オ テ ィク ス の 利 用 が い わ れ て い る2)ことか ら慎 重 な使 用 が 望 ま れ て い 菌 剤 の投 与 につ い て は病 原 菌 を競 合 感 染 源 の1つ タ, る い は下 痢 の 判 定 が た,マ ウ ス,ブ タ る い は抗 生 剤 に よ り

5 7) Zhao T, Doyle MP, Harmon BG, Brown CA, Mueller POE, Parks AH: Reduction of carriage of enterohemorrhagic Escherichia coli 157: H7 in cattle by inoculation with probiotic bacteria. J Clin Microbiol 1998; 36: 641- O ) Fujii J, Kita T, Yoshida S et al.: Direct evidence of neuron impairment by oral infection

6 with verotoxin-producing Escherichia coli 157: H- in mitomycin-treated mice. Infect O Immun 1994; 62: ) Isogai E, Isogai H, Kimura K et al.: Role of tumor necrosis factor alpha in gnotobiotic mice infected with an Escherichia coli O157: H7 strain. Infect Immun 1998; 66: ) Francis DH, Collins JE, Duimstra JR: Infection of Gnotobiotic pigs with an Escherichia coli O157: H7 strain associated with an outbreak of Hemorrhagic colitis. Infect Immun 1986; 51: ) Tzipori S, Wachsmuth IK, Chapman C et al.: The pathogenesis of hemorrhagic colitis caused by Escherichia coli O157: H7 in gnotobiotic piglets. J Infect Dis 1986; 154: ) Beery JT, Doyle MP, Schoeni JL: Colonization of chicken cecae by Escherichia coli associated with hemorrhagic colitis. Appl Environ Microbiol 1985; 49: ) Sueyoshi M, Nakazawa M: Experimental infection of young chicks with attaching and effacing Escherichia coli. Infect Immun 1994; 62: ) Fujii J, Kinoshita Y, Kita T et al.: Magnetic resonance imaging and histopathological study of brain lesions in rabbits given intravenous verotoxin 2. Infect Immun 1996; 64: ) Zoja C, Daniera C, Claudio F et al.: Verotoxin glycolipid receptors determine the localization of microangiopathic process in rabbits given verotoxin-1. J Lab Clin Med 1992; 120: ) Iino H, Fukaya K, Hirasawa Y, Shimizu K, Seo G: Stimulation of bacterial growth of some strains of Bifidobacteriurn by a crude preparation of metabolites from Bacillus rnesentericus TO-A. Biomedical Letters 1993; 48: Estimation of Probiotics by Infection Model of Infant Rabbit with Enterohemorrhagic Escherichia coli O157:H7 Takahiro TACHIKAWA1), Gen-ichiro SEO1), Muneo NAKAZAWA2), Masuo SUEYOSHI3), Tsutomu OH-ISHI4) & Kosuke JOH4) Toa Pharmaceutical Co., Ltd. 1) 2)National Institute of Animal Health 3)Faculty of Agriculture, Miyazaki University 4)The Saitama Children's Medical Center We examined the preventative effect against enterohemorrhagic Escherichia coli O157:H7 (EHEC) infection in rabbits on administration of probiotics contained in Streptococcus faecalis, Clostridium butyricum and Bacillus mesentericus. The probiotics were administered to 5 days old Japanese white rabbits throughout the experiment. Inoculation of 106 colony forming units (CFUs) of EHEC per rabbit was tried 8 days after birth. Three days after EHEC inoculation, in the control group (not administered probiotics), diarrhea was observed in approximately 80% of the rabbits, on the other hand, in the group administered probiotics, rabbits showed diarrhea in 15%. In necropsy, the number of EHEC in the contents of the cecum and the number of attaching and effacing (AE) lesions were fewer in the probiotics group than in the control group. These results suggest that infant rabbits are useful as a colitis model for EHEC, and the probiotics are effective for the prevention of the growth of EHEC in the intestine and for diarrhea.

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