Effect of chicken meat environment on gene expression of Campylobacter jejuni and its relevance to survival in food

Campylobacter is the most frequent foodborne zoonotic bacteria worldwide, with chicken meat being overwhelmingly the most important reservoir for human infections. Control measures implemented at the farm level (i.e., biosecurity or vaccination), which have been successfully applied to limit other pathogens, such as Salmonella, have not been effective in reducing Campylobacter occurrence. Thus, new approaches are needed to fully understand the ecological interactions of Campylobacter with host animals to effectively comprehend its epidemiology. The objective of this study was to analyse longitudinally the gut microbiota composition of Campylobacter-infected and non-infected farms to identify any difference that could potentially be indicative of gut colonization by Campylobacter spp. Differences in the colonization rate and timing were observed at the farms that became positive for Campylobacter jejuni over the investigated time points, even though in positive tests, the occurrence of Campylobacter jejuni gut colonization was not observed before the second week of the life of the birds. Significant differences were observed in the abundances of specific bacterial taxa between the microbiota of individuals belonging to farms that became Campylobacter positive during the study and those who remained negative with particular reference to Bacteroidales and Clostridiales, respectively. Moreover, Campylobacter colonization dramatically influenced the microbiota richness, although to a different extent depending on the infection timing. Finally, a key role of Faecalibacterium and Lactobacillus genera on the Campylobacter microbial network was observed. Understanding the ecology of the Campylobacter interaction with host microbiota during infection could support novel approaches for broiler microbial barrier restoration. Therefore, evidence obtained through this study can be used to identify options to reduce the incidence of infection at a primary production level based on the targeted influence of the intestinal microbiota, thus helping develop new control strategies in order to mitigate the risk of human exposure to Campylobacter by chicken meat consumption.

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The Genome-Sequenced Variant of Campylobacter jejuni NCTC 11168 and the Original Clonal Clinical Isolate Differ Markedly in Colonization, Gene Expression, and Virulence-Associated Phenotypes

Journal of Bacteriology ◽

10.1128/jb.186.23.8159.2004 ◽

2004 ◽

Vol 186 (23) ◽

pp. 8159-8159 ◽

Cited By ~ 1

Author(s):

Erin C. Gaynor ◽

Shaun Cawthraw ◽

Georgina Manning ◽

Joanna K. MacKichan ◽

Stanley Falkow ◽

...

Keyword(s):

Gene Expression ◽

Clinical Isolate ◽

Campylobacter Jejuni

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Significant contribution of the CmeABC Efflux pump in high-level resistance to ciprofloxacin and tetracycline in Campylobacter jejuni and Campylobacter coli clinical isolates

Annals of Clinical Microbiology and Antimicrobials ◽

10.1186/s12941-021-00439-6 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Saeed Sharifi ◽

Bita Bakhshi ◽

Shahin Najar-peerayeh

Keyword(s):

Gene Expression ◽

Point Mutation ◽

Campylobacter Jejuni ◽

Significant Contribution ◽

Efflux Pump ◽

Campylobacter Coli ◽

Rapd Pcr ◽

Resistant Strains ◽

High Level ◽

Level Resistance

Abstract Background Campylobacter resistance to antimicrobial agents is regarded as a major concern worldwide. The aim of this study was to investigate the expression of the CmeABC efflux pump and the RAPD-PCR pattern in drug-resistant Campylobacter isolates. Methods A total of 283 stool specimens were collected from children under the age of five with diarrhea. The minimum inhibitory concentration (MIC) of tetracycline and ciprofloxacin was determined by broth microdilution method and E-test, respectively. Detection of tetracycline and ciprofloxacin determinants was done by amplification of tetO gene and PCR-sequencing of the gyrA gene. The cmeABC transcriptional expression was analyzed by Real-time (RT)-PCR. Clonal correlation of resistant strains was determined by RAPD-PCR genotyping. Results Out of 283 fecal samples, 20 (7.02%) samples were positive for Campylobacter spp. Analysis of duplex PCR assay of the cadF gene showed that 737 and 461 bp amplicons were corresponding to Campylobacter jejuni and Campylobacter coli, respectively. All of the 17 phenotypically tetracycline-resistant Campylobacter isolates harbored the tetO gene. Also, four phenotypically ciprofloxacin-resistant Campylobacter isolates had a point mutation at codon 257 of the gyrA gene (ACA to ATA; Thr > Ile). High-level expression of the cmeA gene was observed in ciprofloxacin-resistant and high-level tetracycline-resistant Campylobacter isolates, suggesting a positive correlation between the cmeA gene expression level and tetracycline resistance level. Moreover, a statistically significant difference was observed in the cmeA gene expression between ciprofloxacin-resistant and ciprofloxacin-susceptible strains, which signifies the crucial contribution of the efflux pump in conferring multiple drug resistance phenotype among Campylobacter spp. RAPD analysis of Campylobacter isolates exhibited 16 different patterns. Simpsone`s diversity index of RAPD-PCR was calculated as 0.85, showing a high level of homogeneity among the population; however, no clear correlation was detected among tetracycline and/or ciprofloxacin resistant isolates. Conclusion Significant contribution of the CmeABC efflux pump in conferring high-level resistance to tetracycline and ciprofloxacin was observed in C. jejuni and C. coli clinical isolates. The resistant phenotype is suggested to be mediated by CmeABC efflux pumps, the tetO gene, and point mutation of the gyrA gene. Genotyping revealed no clonal correlation among resistant strains, indicating distinct evolution of tetracycline and ciprofloxacin resistant genotypes among the isolates.

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Correlation between Genotypic Diversity, Lipooligosaccharide Gene Locus Class Variation, and Caco-2 Cell Invasion Potential of Campylobacter jejuni Isolates from Chicken Meat and Humans: Contribution to Virulotyping

Applied and Environmental Microbiology ◽

10.1128/aem.02269-08 ◽

2009 ◽

Vol 75 (13) ◽

pp. 4277-4288 ◽

Cited By ~ 48

Author(s):

Ihab Habib ◽

Rogier Louwen ◽

Mieke Uyttendaele ◽

Kurt Houf ◽

Olivier Vandenberg ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Genotypic Diversity ◽

Chicken Meat ◽

Pcr Screening ◽

Class A ◽

Invasion Potential ◽

Cell Surface Carbohydrate ◽

Significant Difference ◽

Class B ◽

Class C

ABSTRACT Significant interest in studying the lipooligosaccharide (LOS) of Campylobacter jejuni has stemmed from its potential role in postinfection paralytic disorders. In this study we present the results of PCR screening of five LOS locus classes (A, B, C, D, and E) for a collection of 116 C. jejuni isolates from chicken meat (n = 76) and sporadic human cases of diarrhea (n = 40). We correlated LOS classes with clonal complexes (CC) assigned by multilocus sequence typing (MLST). Finally, we evaluated the invasion potential of a panel of 52 of these C. jejuni isolates for Caco-2 cells. PCR screening showed that 87.1% (101/116) of isolates could be assigned to LOS class A, B, C, D, or E. Concordance between LOS classes and certain MLST CC was revealed. The majority (85.7% [24/28]) of C. jejuni isolates grouped in CC-21 were shown to express LOS locus class C. The invasion potential of C. jejuni isolates possessing sialylated LOS (n = 29; classes A, B, and C) for Caco-2 cells was significantly higher (P < 0.0001) than that of C. jejuni isolates with nonsialylated LOS (n = 23; classes D and E). There was no significant difference in invasiveness between chicken meat and human isolates. However, C. jejuni isolates assigned to CC-206 (correlated with LOS class B) or CC-21 (correlated with LOS class C) showed statistically significantly higher levels of invasion than isolates from other CC. Correlation between LOS classes and CC was further confirmed by pulsed-field gel electrophoresis. The present study reveals a correlation between genotypic diversity and LOS locus classes of C. jejuni. We showed that simple PCR screening for C. jejuni LOS classes could reliably predict certain MLST CC and add to the interpretation of molecular-typing results. Our study corroborates that sialylation of LOS is advantageous for C. jejuni fitness and virulence in different hosts. The modulation of cell surface carbohydrate structure could enhance the ability of C. jejuni to adapt to or survive in a host.

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Comparison of Genotypes and Antibiotic Resistances of Campylobacter jejuni and Campylobacter coli on Chicken Retail Meat and at Slaughter

Applied and Environmental Microbiology ◽

10.1128/aem.00493-13 ◽

2013 ◽

Vol 79 (12) ◽

pp. 3875-3878 ◽

Cited By ~ 13

Author(s):

Sonja Kittl ◽

Bożena M. Korczak ◽

Lilian Niederer ◽

Andreas Baumgartner ◽

Sabina Buettner ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Human Infection ◽

Chicken Meat ◽

Campylobacter Coli ◽

Production Chain ◽

Content Type ◽

Resistance Patterns ◽

Retail Meat ◽

Retail Chicken Meat ◽

Antibiotic Resistance Patterns

ABSTRACTMultilocus sequence typing (MLST) and antibiotic resistance patterns ofCampylobacter jejuniandCampylobacter colifrom retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection.

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Epidemiological investigations onCampylobacter jejuniin households with a primary infection

Journal of Hygiene ◽

10.1017/s002217240006486x ◽

1984 ◽

Vol 93 (2) ◽

pp. 325-332 ◽

Cited By ~ 63

Author(s):

J. Oosterom ◽

C. H. den Uyl ◽

J. R. J. Bänffer ◽

J. Huisman

Keyword(s):

Campylobacter Jejuni ◽

Primary Infection ◽

Index Patient ◽

Chicken Meat ◽

Circumstantial Evidence ◽

Common Source ◽

Household Contacts ◽

Control Subjects ◽

Source Of Infection ◽

Spread Of Infection

SummaryFifty-four Rotterdam patients in which a primary infection withCampylobacter jejunihad been detected (index patients) were compared with 54 control subjects with regard to the consumption and preparation of foods 7 days before onset of illness and the keeping of pet animals. Significantly more index patients than controls had eaten chicken meat (47v. 29;P= 0·0002), particularly at barbecues (14v. 2;P= 0·0015). Marginally more index patients had eaten pork (47v. 39;P= 0·048) or inadequately heated meat (13v. 8), though in the last case numbers were too small to be statistically significant. The consumption of beef or mutton and outdoor eating (other than at barbecues) were essentially the same in both groups. There was no significant association with the keeping of pet animals, although a few more index patients had cage birds than controls (18v. 12).Twenty-one (15%) of 130 household contacts of index patients also suffered from diarrhoea during the same period. Circumstantial evidence pointed to a common source of infection with the index patient in 13 instances (nine households) and probable intrafamilial spread of infection in six instances.Campylobacters were isolated from one of 110 swabs of kitchen work surfaces and eight of 107 swabs taken from lavatory bowls in index households.

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FlhF and Its GTPase Activity Are Required for Distinct Processes in Flagellar Gene Regulation and Biosynthesis in Campylobacter jejuni

Journal of Bacteriology ◽

10.1128/jb.00884-09 ◽

2009 ◽

Vol 191 (21) ◽

pp. 6602-6611 ◽

Cited By ~ 39

Author(s):

Murat Balaban ◽

Stephanie N. Joslin ◽

David R. Hendrixson

Keyword(s):

Gene Expression ◽

Gene Regulation ◽

Campylobacter Jejuni ◽

Regulatory System ◽

Flagellar Gene ◽

Gtpase Activity ◽

Gtp Hydrolysis ◽

Genetic Analyses ◽

Flagellar Biosynthesis ◽

Two Component

ABSTRACT FlhF proteins are putative GTPases that are often necessary for one or more steps in flagellar organelle development in polarly flagellated bacteria. In Campylobacter jejuni, FlhF is required for σ54-dependent flagellar gene expression and flagellar biosynthesis, but how FlhF influences these processes is unknown. Furthermore, the GTPase activity of any FlhF protein and the requirement of this speculated activity for steps in flagellar biosynthesis remain uncharacterized. We show here that C. jejuni FlhF hydrolyzes GTP, indicating that these proteins are GTPases. C. jejuni mutants producing FlhF proteins with reduced GTPase activity were not severely defective for σ54-dependent flagellar gene expression, unlike a mutant lacking FlhF. Instead, these mutants had a propensity to lack flagella or produce flagella in improper numbers or at nonpolar locations, indicating that GTP hydrolysis by FlhF is required for proper flagellar biosynthesis. Additional studies focused on elucidating a possible role for FlhF in σ54-dependent flagellar gene expression were conducted. These studies revealed that FlhF does not influence production of or signaling between the flagellar export apparatus and the FlgSR two-component regulatory system to activate σ54. Instead, our data suggest that FlhF functions in an independent pathway that converges with or works downstream of the flagellar export apparatus-FlgSR pathway to influence σ54-dependent gene expression. This study provides corroborative biochemical and genetic analyses suggesting that different activities of the C. jejuni FlhF GTPase are required for distinct steps in flagellar gene expression and biosynthesis. Our findings are likely applicable to many polarly flagellated bacteria that utilize FlhF in flagellar biosynthesis processes.

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Prevalence and susceptibility to antibiotics from Campylobacter jejuni and Campylobacter coli isolated from chicken meat in southern Benin, West Africa

BMC Research Notes ◽

10.1186/s13104-020-05150-x ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Sylvain Daton Kouglenou ◽

Alidehou Jerrold Agbankpe ◽

Victorien Dougnon ◽

Armando Djiyou Djeuda ◽

Esther Deguenon ◽

...

Keyword(s):

West Africa ◽

Campylobacter Jejuni ◽

Chicken Meat ◽

Campylobacter Coli ◽

Southern Benin

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Two rapid diagnostic procedures for the identification of Campylobacter jejuni/coli in food matrix

Czech Journal of Food Sciences ◽

10.17221/3381-cjfs ◽

2011 ◽

Vol 23 (No. 3) ◽

pp. 121-125

Author(s):

T. Jeleník ◽

Z. Šabatková ◽

K. Demnerová ◽

J. Pazlarová

Keyword(s):

Chronic Illness ◽

Detection Limit ◽

Campylobacter Jejuni ◽

Positive Reaction ◽

Diagnostic Procedures ◽

Chicken Meat ◽

Food Matrix ◽

Selective Enrichment ◽

Campylobacter Species ◽

Campylobacter Spp

Campylobacter species, in particular C. jejuni and C. coli, cause infections which vary in symptoms, ranging from asymptomatic to severe chronic illness. The only ISO method for the detection of Campylobacter spp. until now has been the cultivation by selective enrichment and distinct conditions of growth taking several days to complete. We compared the Singlepath® Campylobacter test which involved 24 h of enrichment in Bolton broth, with PCR-based identification. Chicken meat salad with mayonnaise was spiked with C. jejuni and C. coli and the detection limit was determined. PCR provided the same detection limit of 102 CFU/ml for both strains. The immunotest Singlepath® was positive with C. jejuni only, the quantity of cells being 103 CFU/ml. C. coli was undetectable by Singlepath®, even the concentration of 105 did not reveal a positive reaction.

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Influence of Glycyrrhiza glabra Extract on Growth, Gene Expression of Gut Integrity, and Campylobacter jejuni Colonization in Broiler Chickens

Frontiers in Veterinary Science ◽

10.3389/fvets.2020.612063 ◽

2020 ◽

Vol 7 ◽

Author(s):

Doaa Ibrahim ◽

Alaa H. Sewid ◽

Ahmed H. Arisha ◽

Amir H. abd El-fattah ◽

Adel M. Abdelaziz ◽

...

Keyword(s):

Gene Expression ◽

Body Weight ◽

Campylobacter Jejuni ◽

Broiler Chickens ◽

Body Weight Gain ◽

Control Diet ◽

Tight Junction Proteins ◽

Expression Of Genes ◽

Real Time Qpcr ◽

Junction Proteins

Phytogenic feed additives have been gaining considerable interest due to their ability to improve gut health and thereby performance of broiler chickens. The impact of Glycyrrhiza glabra (licorice) extract (GE) on expression of genes coding for tight junction proteins and gut protection and Campylobacter jejuni colonization in broilers has not been discussed until now. Thus, the current study assessed the effective dose of GE for maximum growth in broiler chickens, clear-cut molecular mechanisms related to integrity and health of intestine, and controlling C. jejuni colonization. Over a 35-day feeding period, a total of 500 Ross broiler chicks were allocated to five groups; the first group was fed a control diet without GE and the second group to the fifth group were fed a control diet with GE (0.25, 0.5, 1, and 2 g/kg of diet); each group comprised 100 chicks with 10 replicates (10 birds/replicate). Birds fed GE had an improved body weight gain and feed conversion ratio. Furthermore, the highest body weight gain was observed in the group that received 1 g/kg of GE (P < 0.05). The expression of genes coding for tight junction proteins [occludin and junctional adhesion molecules (JAM)] was upregulated in all groups supplemented with GE. Moreover, birds fed 1 g/kg of GE exhibited the maximum gene expression of occludin and JAM [0.2 and 0.3 fold change, respectively (P < 0.05)]. In relation to enterocyte protective genes [glucagon-like peptide (GLP-2) and fatty acid-binding protein (FABP-6)], use of GE significantly upregulated expression of GLP-2 gene with 0.8 fold change in 2 g/kg of the GE supplemented group (P < 0.05) while the expression of FABP-6 gene was not affected by GE supplementation (P > 0.05). After challenge with C. jejuni, the expression of mucin (MUC-2) gene was upregulated and the inflammatory markers such as Toll-like receptors (TLR-4) and interleukin (IL-1β) were downregulated with increasing level of supplemented GE (P < 0.05). The mean log10 count of C. jejuni in cecal samples after 7 days post-infection by culture and real-time qPCR was decreased in groups fed GE in a dose-dependent manner (P < 0.05). In addition, the highest reduction of C. jejuni count in cecal samples by culture and real-time qPCR was observed in the group fed 2 g/kg of GE (2.58 and 2.28 log10 CFU/g, respectively). Results from this study suggested that G. glabra extract (1 g/kg) improved growth performance of broiler chickens, as well as influenced the maintenance of intestinal integrity and reduced C. jejuni shedding from infected birds.

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