• Title/Summary/Keyword: C jejuni

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Use of In Vivo-Induced Antigen Technology to Identify In Vivo-Expressed Genes of Campylobacter jejuni During Human Infection

  • Hu, Yuanqing;Huang, Jinlin;Li, Qiuchun;Shang, Yuwei;Ren, Fangzhe;Jiao, Yang;Liu, Zhicheng;Pan, Zhiming;Jiao, Xin-An
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.363-370
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    • 2014
  • Campylobacter jejuni is a prevalent foodborne pathogen worldwide. Human infection by C. jejuni primarily arises from contaminated poultry meats. Genes expressed in vivo may play an important role in the pathogenicity of C. jejuni. We applied an immunoscreening method, in vivo-induced antigen technology (IVIAT), to identify in vivo-induced genes during human infection by C. jejuni. An inducible expression library of genomic proteins was constructed from sequenced C. jejuni NCTC 11168 and was then screened using adsorbed, pooled human sera obtained from clinical patients. We successfully identified 24 unique genes expressed in vivo. These genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, and other processes. We selected six genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results showed that the selected six genes were significantly upregulated in vivo but not in vitro. In short, these identified in vivo-induced genes may contribute to human infection of C. jejuni, some of which may be meaningful vaccine candidate antigens or diagnosis serologic markers for campylobacteriosis. IVIAT may present a significant and efficient method for understanding the pathogenicity mechanism of Campylobacter and for finding targets for its prevention and control.

Predictive model and quantitative microbial risk assessment of enterohemorrhagic Escherichia coli and Campylobacter jejuni in milk (우유에서 장출혈성 대장균과 캠필로박터균의 행동예측 모델 개발 및 정량적 미생물 위해성 평가 연구)

  • Dong, Jiaming;Min, Kyung Jin;Seo, Kun Ho;Yoon, Ki Sun
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.657-668
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    • 2021
  • We prepared the growth and survival models of enterohemorrhagic Escherichia coli (EHEC) and Campylobacter jejuni in milk as a function of temperature and assessed the microbiological risks associated with the consumption of whole milk. EHEC and C. jejuni were not detected in whole milk (n=195) in the retail market. The minimum growth temperature of EHEC in milk was 7℃. The lag time of EHEC in whole milk was longer than that in skim milk. The survival ability of C. jejuni in milk was better at 4℃ than at 10℃. Lower delta values were observed in whole milk than in skim milk, indicating that C. jejuni survived better in skim milk. The probability of foodborne illness from whole milk consumption was 5.70×10-5 for EHEC and 9.86×10-9 for C. jejuni. Sensitivity analysis results show that the market temperature of EHEC and the dose-response model of C. jejuni are correlated with the probability of foodborne illness.

Specific Detection of Enteropathogen Campylobacter jejuni in Food Using a Polymerase Chain Reaction

  • Shin, Soon-Young;Park, Jong-Hyun;Kim, Wang-June
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.184-190
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    • 1999
  • The use of the polymerase chain reaction (PCR) method was described using two sets of primers based on the ceuN gene (JEJ 1 and JEJ 2) which encodes a protein involved in siderophore transport and 16S rRNA gene (pA and pB) for the sensitive and specific detection of enteropathogen Campylobacter jejuni. Six oligonucleotides were utilized in an amplification experiment and PCR products of predicted sizes were generated from whole cells and boiled cell lysates at the same intensity. Two sets of the primer pairs, JEJ and pAB, were specific enough for all C. jejuni strains tested for the direct use of whole cells without DNA extraction or lysis steps. In the PCR using the pAB primer pair, the detection limit, as determined by the ethidium bromide staining of the amplification products on agarose gels, was at the level of $10^1$ bacteria cells or less in both the pure culture and artificially inoculated milk and chicken enrichment samples, whereas the detection limit with the JEJ primer pair was relatively low, i.e. $10^3$ cells or more in the same PCR samples. The PCR method using either a primer JEJ or pAB was both repeatable and specific for the detection of C. jejuni in food. This method is simply completed within 4 h.

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In-silico characterization and structure-based functional annotation of a hypothetical protein from Campylobacter jejuni involved in propionate catabolism

  • Mazumder, Lincon;Hasan, Mehedi;Rus’d, Ahmed Abu;Islam, Mohammad Ariful
    • Genomics & Informatics
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    • v.19 no.4
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    • pp.43.1-43.12
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    • 2021
  • Campylobacter jejuni is one of the most prevalent organisms associated with foodborne illness across the globe causing campylobacteriosis and gastritis. Many proteins of C. jejuni are still unidentified. The purpose of this study was to determine the structure and function of a non-annotated hypothetical protein (HP) from C. jejuni. A number of properties like physiochemical characteristics, 3D structure, and functional annotation of the HP (accession No. CAG2129885.1) were predicted using various bioinformatics tools followed by further validation and quality assessment. Moreover, the protein-protein interactions and active site were obtained from the STRING and CASTp server, respectively. The hypothesized protein possesses various characteristics including an acidic pH, thermal stability, water solubility, and cytoplasmic distribution. While alpha-helix and random coil structures are the most prominent structural components of this protein, most of it is formed of helices and coils. Along with expected quality, the 3D model has been found to be novel. This study has identified the potential role of the HP in 2-methylcitric acid cycle and propionate catabolism. Furthermore, protein-protein interactions revealed several significant functional partners. The in-silico characterization of this protein will assist to understand its molecular mechanism of action better. The methodology of this study would also serve as the basis for additional research into proteomic and genomic data for functional potential identification.

Selective Detection of Campylobacter sp. and Campylobacter jejuni in Meat Food by Polymerase Chain Reaction (PCR을 이용한 육류 내 Campylobacter sp. 및 Campylobacter jejuni의 분리 검출)

  • Joo, Jong-Won;Hong, Kyung-Pyo;Kim, Yong-Hui;Cho, Sang-Buem
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.5
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    • pp.753-759
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    • 2008
  • The principal objective of this study was to develop the optimum oligonucleotide primers for the simple detection of Campylobacter in food samples. In order to achieve this goal, a variety of oligonucleotide primers were designed via the modification of 16S rDNA, ceuE and mapA sequences of Campylobacter. Through the subsequent analysis of the specificity and sensitivity of primers, two types of oligonucleotide primers, CB4 and CJ1, were selected for Campylobacter genus-specific and C. jejuni species-specific primers, respectively. The detection limit was found to be $10^0{\sim}10^1$ cells per reaction with the prepared cell suspension, however, the sensitivity in the meat samples was less, at $10^1{\sim}10^2$. We suggested that PCR inhibitors such as hemoglobin or immunoglobulin in pork or beef influenced.

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Isolation and Antimicrobial Susceptibility of Campylobacter jejuni from Diarrhea Patients (설사환자에서 Campylobacter jejuni의 분리 및 항생제 감수성 특성)

  • Park, Eun-Hee;Kim, Joung-A;Choi, Seung-Hwa;Bin, Jae-Hun;Cheigh, Hong-Sik;Suk, Dong-Hee;Lee, Su-Chul;Kim, Young-Hui
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.811-815
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    • 2007
  • ln this study we isolated 27 isolates of Campylobacter jejuni from stool samples of 882 diarrheal patients. The seasonal distribution of patients was highest at July (11.7%). All the isolates of C. jejuni hydrolyzing sodium hippurate were serotyped on basis of heat-stable antigens, and identified with the use of passive hemagglutination assay. A total of 59.3% among 27 C. jejuni isolates were identified into 6 different serotypes, which serotype HS2, HSl/44, and HS2l were dominant. Antibiotics resistant rates of C. jejuni isolates were shown to be 100%, 63.0%, 51.9%, 37.0%, 33.3%, 25.9% and 7.4% to cephalothin, trimethoprim- sulfamethoxazole, tetracycline, ciprofloxacin, ampicillin, gentamycin and clindamycin, respectively. All isolates were sensitive to the erythromycin and imipenem.

Effects of Freezing and Thawing Treatments on Natural Microflora, Inoculated Listeria monocytogenes and Campylobacter jejuni on Chicken Breast (냉동과 해동처리가 계육 가슴살의 natural microflora, 접종된 Listeria monocytogenes와 Campylobacter jejuni에 미치는 영향)

  • Choi, Eun Ji;Chung, Young Bae;Kim, Jin Se;Chun, Ho Hyun
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.42-50
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    • 2016
  • The effects of freezing and thawing conditions on microbiological quality and microstructure change of inoculated (Listeria monocytogenes and Campylobacter jejuni) and non-inoculated chicken breasts were investigated. Chicken breasts were frozen with air blast freezing (-20, -70, and $-150^{\circ}C$), ethanol ($-70^{\circ}C$) and liquid nitrogen ($-196^{\circ}C$) immersion freezing. There were no significant differences on the populations of L. monocytogenes inoculated with chicken breasts under different freezing conditions. However, air blast freezing ($-20^{\circ}C$) resulted in significant reductions for total aerobic bacteria and C. jejuni compared to the control and other freezing treatments. The frozen samples were thawed with (hot or cold) air blast, water immersion, and high pressure thawing at $4^{\circ}C$ and $25^{\circ}C$. the populations of total aerobic bacteria, and yeast and mold in the frozen chicken breast increased by 5.78 and 4.05 log CFU/g after water immersion thawing ($25^{\circ}C$) treatment. After five freeze-thaw cycles, the populations of total aerobic bacteria, yeast and mold, and C. jejuni were reduced by 0.29~1.40 log cycles, while there were no significant differences (P > 0.05) in the populations of L. monocytogenes depending on the freeze-thaw cycles. In addition, the histological examination of chicken breasts showed an increase in spacing between the muscle fiber and torn muscle fiber bundles as the number of freeze-thaw cycles increased. These results indicate that freezing and thawing processes could affect in the levels of microbial contamination and the histological change of chicken breasts.

Assessment of Inactivation for Campylobacter spp. Attached on Chicken Meat (계육에 오염된 Campylobacter 균의 불활성화 평가)

  • Jang Keum-Il;Jeong Heon-Sang;Kim Chung-Ho;Kim Kwang-Yup
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.302-307
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    • 2005
  • The inactivation efficiency of Campylobacter jejuni were assessed in vitro and in vivo using confocal laser microscopy and flow cytometry. C. jejuni cells were inactivated with $1\%$ (w/v) trisodium phosphate (TSP) and the live cells and inactivated cells were distinguished by staining with LIVE/DEAD BacLight Bacteria Viability fluorescent probe. After treatment of TSP for 5 min, most of C. jejuni cells turned to coccoid form from original spiral shape. C. jejuni cells lost total cell viability in the absence of organic nutrients but did not lost total cell viability in the presence of organic nutrients. In vivo test, C. jejuni cells turned to viable but non-culturable (VBNC) form after TSP treatment and remained alive on chicken skin. C. jejuni cells attached on chicken meat would transform to coccoid form by sanitizer treatment, but could possibly be alive by the benefits of organic nutrients present in chicken meat.

Prevalence of virulence-associated genes and antimicrobial resistance of Campylobacter jejuni from ducks in Gyeongnam Province, Korea

  • Yang, Jung-Wong;Kim, Sang-Hyun;Lee, Woo-Won;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.37 no.2
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    • pp.85-96
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    • 2014
  • Total 99 strains of Campylobacter spp. were isolated from 117 cases of duck's fecal samples. Among 99 strains of Campylobacter spp. isolates, 93 strains (93.9%) were C. jejuni and 6 strains (6.1%) were C. coli. Prevalence of virulence and GBS associated genes of 72 C. jejuni isolates was determined by m-PCR. Among the 10 kinds of virulence associated genes, cadF, dnaJ, flaA and ceuE genes were detected in all of C. jejuni isolates from ducks, racR, pldA, iamA, ciaB, virB11 and docC genes were 87.5%, 84.7%, 77.8%, 48.6%, 13.9% and 11.1%, respectively. Antimicrobial susceptibility test was performed on 72 C. jejuni isolates. The rate of resistance were 62.5% for oxytetracycline, 55.6% for kanamycin, 54.2% for enrofloxacin, 50% for ciprofloxacin, 37.5% for tetracycline and nalidixic acid, 18.1% for ampicillin, 15.3% for streptomycin, and 6.9% for ofloxacin. All isolates were susceptible to erythromycin. The adherence (intracellular and extracellular bacteria) abilities of the 20 isolates to INT-407 cells were between $4.21{\pm}1.27{\times}10^4$ CFU/well and $1.053{\pm}0.451{\times}10^6$ CFU/well from the isolates of cj-55 and cj-52, respectively, and that can be expressed as 0.1033% to 5.2655% to the infecting inoculum. The invasion (intracellular bacteria) abilities of the 20 isolates to INT-407 were between $1.00{\pm}1.73{\times}10^3$ CFU/well and $8.47{\pm}5.16{\times}10^4$ CFU/well from the isolates of cj-13 and cj-47, respectively, and that can be expressed as 0.0050% to 0.4235% to the infecting inoculums. The average CFU/well of 20 campylobacters isolated from ducks for adherence to and invasion were $2.646{\pm}2.886{\times}10^5$ and $3.03{\pm}2.7{\times}10^4$ respectively, and that was $1.3230{\pm}1.2139%$ and $0.1516{\pm}0.1343%$ of the starting viable inoculum. There was considerable correlation ($R^2$=0.627) between the adherence and invasion ability of C. jejuni isolates for INT-407 cell.

Electron Microscopic Studies on the Morphology and Ultrastructures in Campylobacter jejuni treated with Physico-chemical Disinfectants (이화학적 살균제로 처리한 Campylobacter jejuni의 세포 형타와 미세구조에 대한 전자현미경적 연구)

  • Yun, Man-Seok;Oh, Hak-Shik;Kim, Chi-Kyung
    • Applied Microscopy
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    • v.19 no.2
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    • pp.17-26
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    • 1989
  • The cells of Campylobacter jejuni treated with physical or chemical disinfection agents were comparatively examined by scanning and transmission electron microscopies for their morphological features and internal ultrastructures. The normal cells of C. jejuni, showed typical spiral rod shapes. The ribosomes, nucleoids, and other cellular constituents were observed to be distributed evenly throughout the cytoplasm. The cells treated with heat or UV-light were changed to spherical or irregular shapes and their cell envelopes were destroyed to form ghost cells by liberating their cytoplasmic components. The cells treated with chlorine or monochloramine were also changed into irregular round shapes. The chlorinated cells showed very rough surface structures with many blob-like protrusions, while the surface of the monochloramine-treated cells appeared to be relatively smooth.

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