Development of a Selective Agar for Improving Campylobacter jejuni Detection in Food
Jimyeong HaYeongeun SeoYujin KimYukyung ChoiHyemin OhYewon LeeEunyoung ParkJoohyun KangHeeyoung LeeSoomin LeeYohan Yoon
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Abstract Background Campylobacter jejuni is a major gastroenteritis-causing foodborne pathogen. However, it is difficult to isolate when competing bacteria or cold-damaged cells are present. Objective Herein, a medium (Campylobacter selective agar, CSA) was developed and supplemented with catalase, L-serine, L-cysteine, and quercetin for the selective detection of C. jejuni in food. Method The C. jejuni-detection efficiency in media broth and chicken tenders was evaluated. The pathogen was enumerated on modified charcoal–cefoperazone–deoxycholate agar (mCCDA), CSA supplemented with 4 µM catalase (CSA-C4), 8 µM catalase (CSA-C8), 20 mM L-serine (CSA-S20) or 50 mM L-serine (CSA-S50), and mCCDA supplemented with 0.5 mM L-cysteine (mCCDA-LC0.5), 1 mM L-cysteine (mCCDA-LC1), 40 µM quercetin (mCCDA-Q40) or 320 µM quercetin (mCCDA-Q320). The detection efficiency was then evaluated by counting colonies on the selective agar media. Quantitative assessment was also performed using chicken and duck carcasses. Results The C. jejuni detection efficiencies were higher (P < 0.05) in the groups CSA-C4 or CSA-C8, and CSA-S20 or CSA-S50, than mCCDA, and the detection efficiencies were maintained even in the presence of Acinetobacter baumannii, a competing bacterium. In the quantitative test, CSA-C8 and CSA-S50 demonstrated higher C. jejuni-detection efficiencies than mCCDA (control). Conclusions Therefore, CSA-C8 and CSA-S50 improved the detection efficiency of C. jejuni in poultry products by promoting the recovery of cold-damaged cells. Highlights When using CSA-C8 or CSA-S50 developed in this study for detection of C. jejuni in food, detection efficiency was higher than mCCDA.Keywords:
Acinetobacter baumannii
Cefoperazone
Detection and genotyping of Campylobacter jejuni and Campylobacter coli in food samples by fla-DGGE.
The aim of this study was to apply a denaturing gradient gel electrophoresis assay of the flagellin gene(fla-DGGE) for detection and genotyping of Campylobacter jejuni and Campylobacter coli in food samples without prior cμLtivation.RBB(rpeated bead beating),CTAB and acetone-chloroform method were used to extracted DNA of all the samples,and then fla-DGGE was applied.The resμLts of the three methods were the same.10 samples,8 were with Campylobacter jejuni or Campylobacter coli,which 3 samples contaminated by two or more of the Campylobacter jejuni or Campylobacter coli,or their different types.After cloning and sequencing resμLts showed that 7 samples were contaminated with the same Campylobacter jejuni,including 3 samples also have the same type of Campylobacter coli,a serious pollution samples were detected to contain one Campylobacter coli and three types of Campylobacter jejuni.Fla-DGGE method is rapid,accurate,and sensitive and could be used in food in Campylobacter jejuni and Campylobacter coli in the rapid detection and typing.
Campylobacter coli
Flagellin
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Objective To study the drug resistance of the clinical isolates of acinetobacter baumannii. Methods The isolates were identified by API20NE, and the drug sensibility was detected by disk diffusion method. Results Acinetobacter baumannii was more important clinical isolate. The sensibility of imipenem and cefoperazone/sulbactam was 97.7% and 63% respectively. Conclusions The clinical isolates of acinetobacter baumannii were resistant to the more antibiotics.
Acinetobacter baumannii
Cefoperazone
Sulbactam
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OBJECTIVE To investigate the present situation of Acinetobacter baumannii infections and explore the drug resistance to the commonly used antibiotics so as to provide basis for reasonable use of antibiotics.METHODS The clinical specimens obtained from the hospitalized patients between Jun 2011and Jun 2012were cultured for pathogens,then identification and the drug susceptibility testing were performed by using VITEK2Compact fully automatic microorganism analyzer.RESULTS A total of 158strains of A.baumannii were isolated from the clinical specimens,among which 76.58% were isolated from respiratory tract secretions,followed by wound secretions(13.29%).Of all the A.baumannii isolated,27.85% were isolated from the surgical ICU,14.56%from the cadres department,and 10.76% from the respiratory internal medicine department.The A.baumannii strains have produced high drug resistance to the commonly used antibiotics;except for 31.6% to cefoperazone-sulbactam,the drug resistance of the A.baumannii strains varied between 53.8% and 90.5%.Totally 89strains of pandrug-resistant A.baumannii were isolated from the 158strains of A.baumannii,accounting for 56.33%,the drug susceptibility rate to polymyxin E was up to 100.0%,and the drug resistance rate to cefoperazone-sulbactam was 53.6%.CONCLUSIONThe A.baumannii is highly resistant to the commonly used antibiotics,it is necessary to use antibiotics based on the result of drug susceptibility testing.The polymyxin and cefoperazone-sulbactam or sulbactam-containing drugs are preferred for the treatment of pandrug-resistant A.baumannii infections.
Acinetobacter baumannii
Cefoperazone
Sulbactam
Polymyxin B
Polymyxin
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This chapter explores the contribution that population studies have made to one's understanding of the biology of the Campylobacter, and the authors argue that such studies have a central role to play in understanding the epidemiology and pathogenesis of this important group of gram-negative bacteria. Campylobacter jejuni and Campylobacter coli cause the majority of human cases of Campylobacter-associated gastroenteritis; these two organisms are associated with approximately 90 and 10% of cases, respectively. This chapter discusses variation within the genus Campylobacter. Although human infection is one of the most important practical applications of studies of Campylobacter populations, in terms of Campylobacter population dynamics and evolution, infection is probably irrelevant. Understanding the population biology of Campylobacter is, however, crucial in understanding the transmission to humans and developing means for its control. It is instructive to reflect that the first study of C. jejuni and C. coli population structure by multilocus enzyme electrophoresis provided many insights that have proved to be correct and that have been extended and deepened by multilocus sequence typing (MLST) studies. Ongoing nucleotide sequence-based studies involving large numbers of isolates and improved genealogical analysis tools provide the highly attractive prospect that well within the next 10 years, the population biology of these organisms, at least insofar as it relates to human infection, will be effectively resolved.
Multilocus sequence typing
Campylobacter coli
Molecular Epidemiology
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Backgrounds: As zoonotic infectious agents, Campylobacter spp. are important factors causing gastroenteritis in humans. Surveys show that the three strains; Campylobacter jejuni, Campylobacter coli and Campylobacter fetus play a major role in human infections. Identification of these infectious agents is valuable for sanitary control of disease transmission through water resources.
Objectives: The aim of this study was identification and molecular diagnosis of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus in surface waters in Rasht.
Materials and Methods: This cross-sectional study was conducted on 45 samples of surface water in Rasht collected according to water health guidelines. After culture and biochemical tests on collected samples, detection and identification of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus was done using sequence-specific amplification by Multiplex PCR. The results were subjected to statistical analysis using SPSS software.
Results: Out of 45 samples tested, 6 were positive in culture, four of which were identified as Campylobacter jejuni after biochemical tests. Using Multiplex PCR, 8 samples were positive, from which 3 were Campylobacter jejuni, 1 Campylobacter coli and 4 were positive for both Campylobacter jejuni and Campylobacter coli. All the samples did not yield C. fetus.
Conclusions: Multiplex PCR is regarded a diagnostic method with higher sensitivity and specificity than compared to methods for Campylobacter. The prevalence of Campylobacter jejuni and Campylobacter coli in surface waters in Rasht is considerable. Therefore, public health measures for the control of these organisms are recommended.
Campylobacter coli
Campylobacter fetus
Campylobacteriosis
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ObjectiveTo understand the distribution characteristics and drug resistance of Acinetobacter baumannii in clinical specimens.MethodsInfection and drug resistance of A.baumannii in our hospital from 2009 to 2010 was analyzed.352 strains of A.baumannii isolated from patients were identified by MicroScan Neg Combo Panel Type 31 and drug susceptibility tests were performed. Results283 strains(80.4%) of A.baumannii were examined from phlegm.163 strains(46.3%) of A.baumannii were from intensive care unit(ICU).56 strains(15.9%) from neurosurgery.A.baumannii had a higher susceptibility to sulbactam/cefoperazone(95%),and to other cephalosporins antibiotics the resistance rate was more than 60%.ConclusionA.baumannii infection is the main pathogenic bacteria in hospital,especially in the ICU.A.baumanii is forming multiple antibiotic resistance in the process of treatment.Sulbactam/cefoperazone is as first-line antimicrobial agents to treat patients with multiple resistance of A.baumannii infection.
Acinetobacter baumannii
Cefoperazone
Sulbactam
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Campylobacter species, primarily Campylobacter jejuni and Campylobacter coli, are regarded as a major cause of human gastrointestinal disease, commonly acquired by eating undercooked chicken. We describe a PCR-ELISA for the detection of Campylobacter species and the discrimination of C. jejuni and C. coli in poultry samples. The PCR assay targets the 16S/23S ribosomal RNA intergenic spacer region of Campylobacter species with DNA oligonucleotide probes designed for the specific detection of C. jejuni, C. coli, and Campylobacter species immobilized on NucleoLink™ wells and hybridized to PCR products modified with a 5′ biotin moiety. The limit of detection of the PCR-ELISA was 100–300 fg (40–120 bacterial cells) for C. jejuni and C. coli with their respective species-specific oligonucleotide probes and 10 fg (4 bacterial cells) with the Campylobacter genus-specific probe. Testing of poultry samples, which were presumptive positive for Campylobacter following culture on the Malthus V analyzer, with the PCR-ELISA determined Campylobacter to be present in 100% of samples (n = 40) with mixed cultures of C. jejuni/C. coli in 55%. The PCR-ELISA when combined with culture pre-enrichment is able to detect the presence of Campylobacter and definitively identify C. jejuni and C. coli in culture-enriched poultry meat samples.
Campylobacter coli
Oligomer restriction
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1. Campylobacter Infection. 2. Taxonomy, Phylogeny, and Methods for the Identification of Campylobacter Species. 3. Population Genetics of Campylobacter jejuni. 4. Campylobacter jejuni Strain Variation. 5. Advances in Campylobacter jejuni Comparative Genomics Through Whole Genome DNA Microarrays. 6. Prevalence of Campylobacter in the Food and Water Supply: Incidence, Outbreaks, Isolation, and Detection. 7. New Methods for Epidemiological Analysis of Campylobacter jejuni. 8. Plasmids of Campylobacter jejuni 81-176. 9. Mechanisms of Antimicrobial Resistance in Campylobacter. 10. Multidrug Efflux Systems in Campylobacter. 11. Genetic Bases for the Variation in the Lipooligosaccharide Outer Core of Campylobacter jejuni and Possible Association of Glycosyltransferase Genes with Post-Infectious Neuropathies. 12. The Polysaccharide Capsule of Campylobacter jejuni. 13. Protein Glycosylation in Campylobacter. 14. Metabolism, Electron Transport and Bioenergetics of Campylobacter jejuni: Implications for Understanding Life in the Gut and Survival in the Environment. 15. Iron Transport and Regulation. 16. Campylobacter jejuni Stress Responses During Survival in the Food Chain and Colonisation.17. Motility. 18. Campylobacter Chemotaxis. 19. Invasion. 20. Cytolethal Distending Toxin. 21. Interactions of C. jejuni with Non-professional Phagocytic Cells. 22. Campylobacter jejuni Interactions with Professional Phagocytes. 23. Campylobacter spp and the Ability to Elicit Intestinal Inflammatory Responses. 24. Campylobacter jejuni Interaction with Enterocytes - Using Host Gene Expression Analysis to Unravel.
Campylobacter coli
Cytolethal distending toxin
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Campylobacter spp. particularly C. jejuni has been recognized as one of the most prevalent causes of foodborne bacterial illnesses in humans. Most previous studies have focused on the transmission routes of C. jejuni from commercial flock farms to the final retail product. To date, no in vivo studies have addressed the efficacy of sulfadimethoxine in the control of C. jejuni in poultry. This dissertation research proceeds along two lines of investigation. The objectives of the first line of investigation are to determine the enumeration of Campylobacter spp. and the prevalence on both Campylobacter spp. and C. jejuni on live egg shells, to detect the presence and extent of Campylobacter spp. and C. jejuni in live birds raised in battery-cage and cage-free systems and to determine to what extent these bacteria are present in drinking water, feed, enclosures and troughs. The objectives of the second line of investigation are to determine the effects of sulfadimethoxine antibiotic on the enumeration of Campylobacter spp. and the prevalence on both Campylobacter spp. and C. jejuni in broilers and to examine the effects of sulfadimethoxine antibiotic treatments on the enumeration of Campylobacter spp. and prevalence of Campylobacter spp. and C. jejuni from likely sources of cross-contamination to include drinking water, feed, enclosures and troughs. The results from the first line of investigation suggest that the vertical transmission of these bacteria from egg surfaces to newly hatched chicks is not a significant risk factor. Additionally, this study found that the horizontal transmission of Campylobacter spp. and C. jejuni among live broilers is significantly higher (P<0.05) raised in the cage-free system than in the battery-cage system. Prevalence of Campylobacter spp. and C. jejuni in the potential abiotic sources of cross contamination (drinking water, feed, enclosures and troughs) in the cage-free system was significantly higher than in the battery-cage system. The results from the second line of investigation also found that drinking water may be a prime source of Campylobacter spp. and C. jejuni cross contamination. The use of antibiotic sulfadimethoxine can reduce the enumeration and prevalence of Campylobacter spp. and the prevalence of C. jejuni.
Flock
Campylobacteriosis
Campylobacter coli
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We report a new molecular technique for the analysis of Campylobacter jejuni and Campylobacter coli based on the separation of target fragments by denaturing high-performance liquid chromatography(DHPLC).A multiplex PCR(mPCR) assay for detection of Campylobacter jejuni and Campylobacter coli was developed using primers that specifically amplify segments of the 16S rRNA,gyrA and cdt genes.The mPCR had a detection limit of 10 pg/μL genomic DNA for Campylobacter jejuni and 1 pg/μL for Campylobacter coli.No cross-reaction was detected with non-thermo-tolerant Campylobacter and other 22 strains tested.The assay could detect 1.5 CFU/mL Campylobacter jejuni and Campylobacter coli in artificial inoculated chicken meat sample after enrichment at 42 ℃ in microaerophilic condition for 24 h.Test on 172 chicken meat samples detected 18 Campy-lobacter jejuni positive and 7 Campylobacter coli positive.These results indicated that the multiplex PCR-DHPLC assay could be used for specific detection of Campylobacter jejun and Campylobacter coli.
Campylobacter coli
Multiplex
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