• 제목/요약/키워드: C jejuni

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훈제오리에서 캠필로박터균 생물막 및 Viable But Non-Culturable(VBNC) 상태에서의 행동특성 (Behavior of Campylobacter jejuni Biofilm Cells and Viable But Non-Culturable (VBNC) C. jejuni on Smoked Duck)

  • 조혜진;전혜리;윤기선
    • 한국식품영양과학회지
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    • 제45권7호
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    • pp.1041-1048
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    • 2016
  • 본 연구에서 가금류의 주요 병원성 식중독 균을 인위적으로 오염시킨 훈제오리육을 진공포장 조건에서 10, 15, $24^{\circ}C$에 저장하면서 유통기한 동안 관찰한 미생물의 증식 및 생존 결과 Campylobacter jejuni는 저장기간 이내에 사멸하는 경향을 보였으며, Salmonella Typhimurium과 Listeria monocytogenes는 균주의 성장 속도에는 차이가 있었으나 증식하는 경향을 보였다. 훈제오리의 유통온도는 $10^{\circ}C$이며 유통기한이 약 30일인 것을 고려했을 때, 초기 오염 수준이 Campylobacter 균주에 의한 식중독을 유발하게 되는 균수 500 CFU/g 수준 이하에서는 유통기한 내에 문제가 없을 것으로 생각한다. 그러나 낮은 온도에서 저항성이 증가하며 살아있으나 배양은 불가능한 상태인 VBNC 상태의 C. jejuni의 특성에 따라 적절한 조건에서 회복되어 병원성을 일으킬 가능성이 있으므로 C. jejuni에 대한 지속적인 관리가 필요하다. 또한, S. Typhimurium과 L. monocytogenes의 경우 일반적인 유통/보관 온도인 $10^{\circ}C$에서도 증식이 가능하며, 특히 가공품 및 RTE 식품은 적절한 가열처리 없이 소비할 경우 식중독 발생 가능성이 높다는 점에서 제품 제조 단계에서부터 위생적인 관리가 필요하다. 본 연구에서는 C. jejuni biofilm cells을 인위적으로 오염시킨 훈제오리육을 진공포장 하여 일반 유통/보관 온도인 $10^{\circ}C$와 실온, 그리고 일반적으로 C. jejuni가 증식 가능한 온도인 $36^{\circ}C$에서 저장하였으나, C. jejuni biofilm cell은 훈제오리에서는 모든 온도에서 재증식이 불가능한 것으로 관찰되었다. 또한, $10^{\circ}C$의 저온에서 유도한 VBNC 상태의 C. jejuni를 훈제오리에 인위적으로 오염시키고 혐기적 조건에서 $42^{\circ}C$에 1일간 저장하며 VBNC 상태의 C. jejuni의 재증식 가능성을 분석하였으나, 최적 증식 온도인 $42^{\circ}C$에서도 재증식은 관찰되지 않았다. 이처럼 본 연구에서는 biofilm을 형성한 C. jejuni도 VBNC 상태의 C. jejuni는 살아 있으나 훈제오리에서의 증식은 관찰되지 않았다. 따라서 훈제오리에서의 C. jejuni의 위험성은 매우 적은 것으로 생각한다. 그러나 C. jejuni의 경우 매우 적은 양으로도 식중독을 일으킬 수 있고 C. jejuni biofilm 및 VBNC의 특성에 따라 잠재적인 위험성을 포함하는 동시에 유통/보관 온도인 냉장 온도에서 더 잘 살아남는다는 점에서 식중독 발생의 주요 원인으로 작용할 수 있는 교차오염과 전이를 예방하는 것이 중요하므로 이에 대한 관리가 강조되어야 할 것으로 생각한다.

ISOLATION OF Campylobacter jejuni AND C. coli FROM DOMESTIC AND EXPERIMENTAL ANIMALS AND THEIR DRUG SUSCEPTIBILITY

  • Nakai, Y.;Kimura, K.;Sato, M.;Inamoto, T.;Ogimoto, K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제7권4호
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    • pp.505-507
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    • 1994
  • A total of 526 domestic and experimental animals in Miyagi prefecture, Japan were investigated for fecal carriage of Campylobacter jejuni and Campylobacter coli. C. jejuni was detected in chickens (8.2%), dogs (6.3%), pigs (4.3%), cattle (1.8%) and hamsters (1.4%). C. coli was only detected from pigs (20.7%). Drug susceptibility test was performed on 5 strains of C. jejuni isolated from chickens and 13 strains of C. coli isolated from pigs to tylosin (TS), thianphenicol (TP), carbadox (CDX), chroltetracyclin (CTC), vancomycin (VCM), cefoperazone (CPZ), latamoxef (LMOX), GM were highly effective and CTC, CP and PL were moderately effective against both C. jejuni and C. coli. TS and TPH were moderately effective against C. jejuni; however, they were less effective to C. coli. One strain of C. jejuni against CTC considered to be drug resistant. The results suggest that C. jejuni and C. coli can be controlled by several drugs effectively, although a drug resistant strain exists.

Inverse Correlation between Extracellular DNase Activity and Biofilm Formation among Chicken-Derived Campylobacter Strains

  • Jung, Gi Hoon;Lim, Eun Seob;Woo, Min-Ah;Lee, Joo Young;Kim, Joo-Sung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • 제27권11호
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    • pp.1942-1951
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    • 2017
  • Campylobacter jejuni and Campylobacter coli are important foodborne pathogenic bacteria, particularly in poultry meat. In this study, the presence of extracellular DNase activity was investigated for biofilm-deficient Campylobacter strains versus biofilm-forming Campylobacter strains isolated from chickens, to understand the relationship between extracellular DNase activity and biofilm formation. A biofilm-forming reference strain, C. jejuni NCTC11168, was co-incubated with biofilm non-forming strains isolated from raw chickens or their supernatants. The biofilm non-forming strains or supernatants significantly prohibited the biofilm formation of C. jejuni NCTC11168. In addition, the strains degraded pre-formed biofilms of C. jejuni NCTC11168. Degradation of C. jejuni NCTC11168 biofilm was confirmed after treatment with the supernatant of the biofilm non-forming strain 2-1 by confocal laser scanning microscopy. Quantitative analysis of the biofilm matrix revealed reduction of extracellular DNA (16%) and proteins (8.7%) after treatment. Whereas the biofilm-forming strains C. jejuni Y23-5 and C. coli 34-3 isolated from raw chickens and the C. jejuni NCTC11168 reference strain showed no extracellular DNase activity against their own genomic DNA, most biofilm non-forming strains tested, including C. jejuni 2-1, C. coli 34-1, and C. jejuni 63-1, exhibited obvious extracellular DNase activities against their own or 11168 genomic DNA, except for one biofilm non-former, C. jejuni 22-1. Our results suggest that extracellular DNase activity is a common feature suppressing biofilm formation among biofilm non-forming C. jejuni or C. coli strains of chicken origin.

Campylobacter jejuni의 groEL 유전자 산물의 대장균에서의 Chaperon효과 (Chaperon Effects of Campylobacter jejuni groEL Genes Products in Escherichia coli)

  • 임채일;김치경;이길재
    • 미생물학회지
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    • 제32권1호
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    • pp.47-52
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    • 1994
  • Campylobacter jejuni에 48${\circ}C$의 열충격을 30분간 주었을 때, HSP90, HSP66, HSP60의 열충격 단백질들이 합성되었고, 이 단백질들은 각각 E. coli의 hsp87, HSP66 (DnaK), HSP58(GroEL)에 상응하는 단백질들이었다. 여러가지의 제한효소로 처리한 C. jejuni의 chromosomal DNA에 E. coli의 groEL(4.0kb)을 probe로 사용하여 Southern hybridization한 결과, 이들과 상동성을 가지는 유전자들이 있음을 확인하였다. C. jejuni의 groEL 유전자를 pWE15 cosmid를 이용하여 recombinant plasmid pLC1을 만들고, 이를 E. coli B178 groEL44 ts mutant에 형질전환시켜 E. coli LC1을 얻었다. 이 pLC1에는 groEL 유전자가 존재하는 5.7kb인 insert DNA가 포함되어 있었고, 그로부터 subcloning한 pLC101에는 groEL을 포함하는 4.0kb의 DNA가 삽입되어 있었다. 이 recombinant plasmid들이 형질전환된 E. coli LC1과 LC101 균주에서는 C. jejuni의 GroEL 단백질이 과다 생산되었다. C. jejuni의 groEL이 cloning된 E. coli LC1은 42${\circ}C$에서의 생장능력이 회복되었고, ${\lambda}$ vir phage에 대한 감수성도 회복되는 등의 chaperon 효과가 입증되었다.

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Quantitative microbial risk assessment of Campylobacter jejuni in jerky in Korea

  • Ha, Jimyeong;Lee, Heeyoung;Kim, Sejeong;Lee, Jeeyeon;Lee, Soomin;Choi, Yukyung;Oh, Hyemin;Yoon, Yohan
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권2호
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    • pp.274-281
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    • 2019
  • Objective: The objective of this study was to estimate the risk of Campylobacter jejuni (C. jejuni) infection from various jerky products in Korea. Methods: For the exposure assessment, the prevalence and predictive models of C. jejuni in the jerky and the temperature and time of the distribution and storage were investigated. In addition, the consumption amounts and frequencies of the products were also investigated. The data for C. jejuni for the prevalence, distribution temperature, distribution time, consumption amount, and consumption frequency were fitted with the @RISK fitting program to obtain appropriate probabilistic distributions. Subsequently, the dose-response models for Campylobacter were researched in the literature. Eventually, the distributions, predictive model, and dose-response model were used to make a simulation model with @RISK to estimate the risk of C. jejuni foodborne illness from the intake of jerky. Results: Among 275 jerky samples, there were no C. jejuni positive samples, and thus, the initial contamination level was statistically predicted with the RiskUniform distribution [RiskUniform (-2, 0.48)]. To describe the changes in the C. jejuni cell counts during distribution and storage, the developed predictive models with the Weibull model (primary model) and polynomial model (secondary model) were utilized. The appropriate probabilistic distribution was the BetaGeneral distribution, and it showed that the average jerky consumption was 51.83 g/d with a frequency of 0.61%. The developed simulation model from this data series and the dose-response model (Beta Poisson model) showed that the risk of C. jejuni foodborne illness per day per person from jerky consumption was $1.56{\times}10^{-12}$. Conclusion: This result suggests that the risk of C. jejuni in jerky could be considered low in Korea.

Ethanol 충격에 의한 Campylobacter jejuni 의 Ethanol 내성 (Ethanol Tolerance of Campylobacter jejuni by Ethanol Shock)

  • 김치경;가익현
    • 미생물학회지
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    • 제30권5호
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    • pp.377-382
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    • 1992
  • Ethanol 충격에 의한 Campylobacter jejuni 의 생존성 및 ethanol 충격 단백질의 합성과 내성반응을 연구하였다. 1% 의 ethanol 충격에 대해서는 60분, 3% 의 ethanol 충격에서는 30분, 5%의 ethanol 충격에서는 10분대데, 분자량이 90, 66, 60, 45, 24 kd 인 충격 단백질이 합성되는 것을 autoradiography 로 확인하였다. C. jejuni 를 1% 와 3% 의 ethanol 에서 30 분간 충격을 준후 각각 3% 와 5% 의 농도에 노출시켰을때의 생존율은 ethanol 충격엾이 직접 3% 와 5% 의 ethanol에 노출시켰을 때보다 $10^{1}$ $-10^{2}$ 정도 높게 나타났다. 또 5% 에서 10분간 충격을 준후7% 의 ethanol 에 노출시켰을 때에도 직접 7% 에 노출시켰을 때보다 생존율이 $10^{2}$ 정도 높게 나타났다. 그러므로 1-5% 의 ethanol 로 충격을 받은 C. jejuni 는 그보다 높은 농도의 ethanol 에 대하여 ethanol 충격단백질에 의한 내성반응이 나타나 생존율의 감소가 훨씬 적어졌다.

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Campylobacter jejuni에서 고온충격 단백질의 합성과 내열성 (Synthesis and thermotolerance of heat shock proteins in campylobacter jejuni)

  • 김치경;김현옥;이길재
    • 미생물학회지
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    • 제29권1호
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    • pp.49-55
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    • 1991
  • The heat shock responses of Campylobacter jejuni were studied by examination of their survival rates and synthesis of heat shocd proteins. When C. jejuni cells were treated at the sublethal temperatures of 48.deg.C for 30 minutes, most of the cells maintained their viabilities and synthesized the heat shock proteins of 90, 73, and 66 kD in molecular weight. By the method of two-dimensional electrophoresis, the heat shock proteins of C. jejuni were identified to be Hsp90, Hsp73, and Hsp66. During the heat shock at 48.deg.C, the heat shock proteins were induced from about 5 minutes after the heat shock treatment. Their synthesis was continued upto 30 minutes, but remarkably retarded after 50 minutes. When C. jejune cells were heat shocked at 51.deg.C for 30 minutes, the survival rates of the cells were decreased by about $10^{3}$ fold and synthesis of heat shock proteins and normal proteins was also generally retarded. The cells exposed to 55.deg.C for 30 minutes died off by more than $10^{5}$ cells and the new protein synthesis was not observed. But when C. jejuni cells were heat-shocked at the sublethal temperature of 48.deg.C for 15 to 20 minutes and then were exposed at the lethal temperature of 55.deg.C for 30 minutes, their viabilities were higher than those exposed at 55.deg.C for 30 minutes without pre-heat shock at 48.deg.C. Therefore, the heat shock proteins synthesized at the sublethal temperature of 48.deg.C in C. jejuni were thought to be responsible for thermotolerance. However, when C. jejuni cells heat-shocked at various ranges of sublethal and lethal temperatures were placed back to the optimum temperature of 42.deg.C, the multiplication patterns of the cells pretreated at different temperatures were not much different each other.

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Role of flgA for Flagellar Biosynthesis and Biofilm Formation of Campylobacter jejuni NCTC11168

  • Kim, Joo-Sung;Park, Changwon;Kim, Yun-Ji
    • Journal of Microbiology and Biotechnology
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    • 제25권11호
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    • pp.1871-1879
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    • 2015
  • The complex roles of flagella in the pathogenesis of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are important. Compared with the wild-type, an insertional mutation of the flgA gene (cj0769c) demonstrated significant decrease in the biofilm formation of C. jejuni NCTC11168 on major food contact surfaces, such as polystyrene, stainless steel, and borosilicate glass. The flgA mutant was completely devoid of flagella and non-motile whereas the wild-type displayed the full-length flagella and motility. In addition, the biofilm formation of the wild-type was inversely dependent on the viscosity of the media. These results support that flagellar-mediated motility plays a significant role in the biofilm formation of C. jejuni NCTC11168. Moreover, our adhesion assay suggests that it plays an important role during biofilm maturation after initial attachment. Furthermore, C. jejuni NCTC11168 wild-type formed biofilm with a net-like structure of extracellular fiber-like material, but such a structure was significantly reduced in the biofilm of the flgA mutant. It supports that the extracellular fiber-like material may play a significant role in the biofilm formation of C. jejuni. This study demonstrated that flgA is essential for flagellar biosynthesis and motility, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

Campylobacter jejuni 의 열충격 반응과 그유전자에 관한 연구

  • 김치경;임채일;이길재
    • 미생물학회지
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    • 제30권3호
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    • pp.232-238
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    • 1992
  • Campylobacter jejuni 에 열처리를 했을 때 그들의 생존성 및 열충격 단백질 합성의 양상과 더불어, dnaK 와 groESL 유전자를 이용하여 C. jejuni 의 열충격 유전자를 검출하여 그 특성을 E. coli 의 열충격 유전자와 비교하였다. C. jejuni 의 열충격 단백질은 48.deg.C 에서 가장 잘 발형되었으며, 48.deg.C 에서 30 분간의 처리중 세포들의 생존율은 떨어지지 않았다. C. jejuni 의 열충격 단백질로서의 Hsp90, Hsp66, Hsp60 이 합성되는 것을 SDS-PAGE 및 방사선사진법을 통해 확인하였다. dnaK 와 groESL 을 DNA 탐침자로 이용하여 Southern hybridization 한 결과, C. jejuni 의 열충격 유전자도 groESL 과 dnaK 유전자와 상동성을 가진 염기서열을 가지고 있었으나, 두 균주사이에는 열충격유전자를 내포하고 있는 DNA 상에서 제한효소의 절단부위에 차이가 있었다.

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Role of eptC in Biofilm Formation by Campylobacter jejuni NCTC11168 on Polystyrene and Glass Surfaces

  • Lim, Eun Seob;Kim, Joo-Sung
    • Journal of Microbiology and Biotechnology
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    • 제27권9호
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    • pp.1609-1616
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    • 2017
  • The complex roles of cell surface modification in the biofilm formation of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are poorly understood. In a screen of mutants from random transposon mutagenesis, an insertional mutation in the eptC gene (cj0256) resulted in a significant decrease in C. jejuni NCTC11168 biofilm formation (<20%) on major food contact surfaces, such as polystyrene and borosilicate glass, when compared with wild-type cells (p < 0.05). In C. jejuni strain 81-176, the protein encoded by eptC modified cell surface structures, such as lipid A, the inner core of lipooligosaccharide, and the flagellar rod protein (FlgG), by attaching phosphoethanolamine. To assess the role of eptC in C. jejuni NCTC11168, adherence and motility tests were performed. In adhesion assays with glass surfaces, the eptC mutant exhibited a $0.77log\;CFU/cm^2$ decrease in adherence compared with wild-type cells during the initial 2 h of the assay (p < 0.05). These results support the hypothesis that the modification of cell surface structures by eptC affects the initial adherence in biofilm formation of C. jejuni NCTC11168. In motility tests, the eptC mutant demonstrated reduced motility when compared with wild-type cells, but wild-type cells with the transposon inserted in a gene irrelevant to biofilm formation (cj1111c) also exhibited decreased motility to a similar extent as the eptC mutant. This suggests that although eptC affects motility, it does not significantly affect biofilm formation. This study demonstrates that eptC is essential for initial adherence, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.