Key words =: Helicobacter pylori, viable but nonculturable
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1 Key words =: Helicobacter pylori, viable but nonculturable
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3 Fig. 1 Phase contrast microscopy dium. left : before incubation, Fig. 2 Molecular RAPD method. C: typing Lane M: of Helicobacter Marker 態 のH.pylori of Helicobacter pylori in supplemented right: pylori incubate by (20 bp ladder), H. pylori ATCC 43504, 1 : Source of strain, 17 liquid me- after 3 days. Fig. 3 Growth curve of Helicobacter pylori in liquid medium 2 : After resuscitation. な い ま で に 傷 害 を 受 け た細 胞 で あ る こ と を 示 唆 し 考 て い る.お そ ら く,ヒ 察 態 の 細 胞 は,培 養 出 来 な い まで に傷 害 を トは現 在 知 られ て い る どの 培 地 よ り も良 好 な,VNCの 回 復 環 境 と考 え られ て 受 け た細 胞 と考 え られ る が15),そ の よ うな 細 胞 は い る16).そ の 菌 が 生 きて い る か ど うか は,最 終 的 選択 分 離 の 条 件 下 で は 発 育 し な い が,そ に培 養 が で き る こ とが 証 明 さ れ て は じめ て 決 定 で の条件 を 除去 す る か,ま た は傷 害 を修 復 させ る よ う な 方 法 き るが,coccoid を講 ず れ ば,発 育 性 が 回 復 す る と され る.細 胞 修 発 育 可能 な ラ セ ン状 桿 菌 へ と復 元 され た事 実 は, 復 剤 を加 えた 培 地 中 で の み,coccoid formか らの 発 育 が 認 め られ た事 実 は,人 工 培 地 中 で 発 育 出 来 平 成11年1月20日 formのh.pyloriが,人 工 培地 で 態 で あ る 証 明 とな っ た.休 眠 状 態 の 結 核 菌 は発 育 し な い が 代 謝 活 性 は 持 っ て い る17)との 報 告
4 Table 1 Growth of Helicobacter pylori in supplemented medium 1) Warren JR, Marshall BJ : Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet 1983 ; 321: ) Bode G, Mauch F, Malfertheiner P : The coccoid form of Helicobacter pylori,criteria for their viability. Epidemiol Infect 1993; 111 : ) Roszak DB, Grimes DJ, Colwell RR : Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems. Can J Microbiol 1984 ; 30 : ) Roszak DB, Clowell RR : Survival strategies of bacteria in the natural environment. Microbiol Rev 1987; 51: ) Shahamat M, Mai U, Paszko-Kolva C. Kessel M, Clowell RR : Use of autoradiography to assess of viability of Helicobacter pylori in water. Appl Environ Microbiol 1993; 59: ) Rollins DM, Clowell WW : Viable but nonculturable state of Campylobacter jejuni and it's role in survival in the natural aquatic environment. Appl Environ Microbiol 1986; 52: ) Xu H-S, Roberts N, Singleton FL, Attwell RW,
5 Grimes DL, Colwell RR : Survival and viability of nonculturable Escherichia coli and Vibrio cholerae in the estuarine and marine environment. Microb Ecol 1982; 8 : ) Nilsson L, Oliver JD, Kjelleberg S : Resuscitation of Vibrio vulnificus from the viable but nonculturable state. J Bacteriol 1991 ; 173 : ) Jones DM, Sutcliffe EM, Curry A : Recovery of viable but nonculturable Campylobacter jejuni.j Gen Microbiol 1991 ; 137: ) Colwell RR, Brayton BR, Grimes DJ, Roszak DB, Huq SA, Palmer LM : Viable but nonculturable Vibrio cholerae and related pathogens in the environment : implications for release of genetically engineered microorganisms. Biotechnol 1985 ; 3 : ) Enroth H, Engstrand L : Immunomagnetic separation and PCR for detection of Helicobacter pylori in water and stool specimens. J Clin Microbiol 1995 ; 33 : ) Wai SN, Moriya T, Kondo K, Misumi H, Amako K : Resuscitation of Vibrio cholerae 0 1 strain TSI- 4 from a viable but nonculturable state by heat shock. FEMS Microbiol Lett 1996 ; 136 : ) Welsh J, McClelland M : Characterization of pathogenic microorganisms by genomic fingerprinting using arbitrarily primes PCR. Persing, D. H. (Eds), Diagnostic molecular microbiology, principles and applications. ASM press, Washington D.C., 1993, ) Weichart D, Kjelleberg S : Stress resistance and recovery potential of culturable and viable but nonculturable cells of Vibrio vulnificus.microbiology 1996 ; 142 : ) Colwell RR, Brayton P, Herrington D, Tall B, Huq A, Levine MM : Viable but nonculturable Vibrio cholerae 0 1 revert to a culturable state in the human intestine. World J Microbiol Biotechonol 1996 ; 12 : ) Wayne LG : Dormant of Mycobacterium tuberculosis and latency of disease. Eur J Clin Microbiol Infect Dis 1994 ; 13 : ) Nilsson L, Oliver JD, Kjelleberg S Resuscitation of Vibrio vulnificus from the viable but nonculturable state. J Bacteriol 1991 ; 173 : Resuscitation from the Viable but Nonculturable State of Helicobacter pylori Manabu KUROKAWA1), Masafumi NUICINA1), Hisao NAKANISHI1), Syusuke TOMITA', Toshihide TAMURA3) & Takashi SHIMOYAMA4) 1)Department of Bacteriology, Kobe Institute of Health 2)Department of Internal Medicine of Gastroenterology, Kobe City General Hospital 3)Department of Bacteriology, Hyogo College of Medicine 4)Department of Internal Medicine of Fourth, Hyogo College of Medicine It is well known that the change of state from the culturable to not culturable on pathogenic organisms could easily occurred. These states of bacteria were so called viable but nonculturable (VNC). The coccoid form of H. pylori is seems to be in this state. The possibility of resuscitation of H. pylori was examined in vitro. The coccoid form bacteria was treated with Ammonium Sulfate and heat shock before culture, and cultured in Brucella broth containing, sodium pyruvate, laked human erythrocyte and serum. The growth of the rod spiral form was found in the coccoid form bacteria, and further the bacteria could grow on blood agar and Skirrow agar as well as the original strain. It was strongly suggested that the resuscitation from VNC state of cells to culturable form consists of two processes, stimulation and supplemented of appropriate nutrition.
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