scholarly journals Transcriptomic analysis of caecal tissue in inbred chicken lines that exhibit heritable differences in resistance to Campylobacter jejuni

2021 ◽  
Author(s):  
Kay M. Russell ◽  
Jacqueline Smith ◽  
Abi Bremner ◽  
Cosmin Chintoan-Uta ◽  
Lonneke Vervelde ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Post Infection ◽  
Cell Signalling ◽  
Transcriptional Response ◽  
Differential Resistance ◽  
Poultry Meat ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Source Of Infection ◽  
Bacterial Gastroenteritis

Abstract Background Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans and the handling or consumption of contaminated poultry meat is a key source of infection. Selective breeding of poultry that exhibit elevated resistance to Campylobacter is an attractive control strategy. Here we studied the global transcriptional response of inbred chicken lines that differ in resistance to C. jejuni colonisation at a key site of bacterial persistence. Results Three-week-old chickens of line 61 and N were inoculated orally with C. jejuni strain M1 and caecal contents and tonsils were sampled at 1 and 5 days post-infection. Caecal colonisation was significantly lower in line 61 compared to line N at 1 day post-infection, but not 5 days post-infection. RNA-Seq analysis of caecal tonsils of both lines revealed a limited response to C. jejuni infection compared to age-matched uninfected controls. In line N at days 1 and 5 post-infection, just 8 and 3 differentially expressed genes (DEGs) were detected (fold-change > 2 and false-discovery rate of < 0.05) relative to uninfected controls, respectively. In the relatively resistant line 61, a broader response to C. jejuni was observed, with 69 DEGs relating to immune regulation, cell signalling and metabolism at 1 day post-infection. However, by day 5 post-infection, no DEGs were detected. By far, the greatest number of DEGs were between uninfected birds of the two lines implying that differential resistance to C. jejuni is intrinsic. Of these genes, several Major Histocompatibility Complex class I-related genes (MHCIA1, MHCBL2 and MHCIY) and antimicrobial peptides (MUC2, AvBD10 and GZMA) were expressed to a greater extent in line N. Two genes within quantitative trait loci associated with C. jejuni colonisation were also more highly expressed in line N (ASIC4 and BZFP2). Quantitative reverse-transcriptase PCR analysis of a subset of transcripts confirmed the RNA-Seq results. Conclusions Our data indicate a limited transcriptional response in the caecal tonsils of inbred chickens to intestinal colonisation by Campylobacter but identify a large number of differentially transcribed genes between lines 61 and N that may underlie variation in heritable resistance to C. jejuni.

scholarly journals Transcriptomic analysis of caecal tissue in inbred chicken lines that exhibit heritable differences in resistance to Campylobacter jejuni

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Kay M. Russell ◽  
Jacqueline Smith ◽  
Abi Bremner ◽  
Cosmin Chintoan-Uta ◽  
Lonneke Vervelde ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Post Infection ◽  
Cell Signalling ◽  
Transcriptional Response ◽  
Differential Resistance ◽  
Poultry Meat ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Source Of Infection ◽  
Bacterial Gastroenteritis

Abstract Background Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans and the handling or consumption of contaminated poultry meat is a key source of infection. Selective breeding of poultry that exhibit elevated resistance to Campylobacter is an attractive control strategy. Here we studied the global transcriptional response of inbred chicken lines that differ in resistance to C. jejuni colonisation at a key site of bacterial persistence. Results Three-week-old chickens of line 61 and N were inoculated orally with C. jejuni strain M1 and caecal contents and tonsils were sampled at 1 and 5 days post-infection. Caecal colonisation was significantly lower in line 61 compared to line N at 1 day post-infection, but not 5 days post-infection. RNA-Seq analysis of caecal tonsils of both lines revealed a limited response to C. jejuni infection compared to age-matched uninfected controls. In line N at days 1 and 5 post-infection, just 8 and 3 differentially expressed genes (DEGs) were detected (fold-change > 2 and false-discovery rate of < 0.05) relative to uninfected controls, respectively. In the relatively resistant line 61, a broader response to C. jejuni was observed, with 69 DEGs relating to immune regulation, cell signalling and metabolism at 1 day post-infection. However, by day 5 post-infection, no DEGs were detected. By far, the greatest number of DEGs were between uninfected birds of the two lines implying that differential resistance to C. jejuni is intrinsic. Of these genes, several Major Histocompatibility Complex class I-related genes (MHCIA1, MHCBL2 and MHCIY) and antimicrobial peptides (MUC2, AvBD10 and GZMA) were expressed to a greater extent in line N. Two genes within quantitative trait loci associated with C. jejuni colonisation were also more highly expressed in line N (ASIC4 and BZFP2). Quantitative reverse-transcriptase PCR analysis of a subset of transcripts confirmed the RNA-Seq results. Conclusions Our data indicate a limited transcriptional response in the caecal tonsils of inbred chickens to intestinal colonisation by Campylobacter but identify a large number of differentially transcribed genes between lines 61 and N that may underlie variation in heritable resistance to C. jejuni.

scholarly journals Bridging the Gap: A Role for Campylobacter jejuni Biofilms

2020 ◽  
Vol 8 (3) ◽  
pp. 452 ◽  
Author(s):  
Greg Tram ◽  
Christopher J. Day ◽  
Victoria Korolik
Keyword(s):  
Life Cycle ◽  
Campylobacter Jejuni ◽  
Biofilm Formation ◽  
Intestinal Tract ◽  
Food Preparation ◽  
Poultry Meat ◽  
Intrinsic Factors ◽  
Human Host ◽  
Bacterial Gastroenteritis ◽  
Developed World

Campylobacter jejuni is the leading cause of bacterial gastroenteritis in the developed world. Cases of Campylobacteriosis are common, as the organism is an avian commensal and is passed on to humans through contaminated poultry meat, water, and food preparation areas. Although typically a fastidious organism, C. jejuni can survive outside the avian intestinal tract until it is able to reach a human host. It has long been considered that biofilms play a key role in transmission of this pathogen. The aim of this review is to examine factors that trigger biofilm formation in C. jejuni. A range of environmental elements have been shown to initiate biofilm formation, which are then affected by a suite of intrinsic factors. We also aim to further investigate the role that biofilms may play in the life cycle of this organism.

scholarly journals Kinetics of the Cellular and Transcriptomic Response to Eimeria maxima in Relatively Resistant and Susceptible Chicken Lines

2021 ◽  
Vol 12 ◽  
Author(s):  
Abi Bremner ◽  
Sungwon Kim ◽  
Katrina M. Morris ◽  
Matthew John Nolan ◽  
Dominika Borowska ◽  
...  
Keyword(s):  
Post Infection ◽  
Differential Resistance ◽  
Poultry Production ◽  
Rna Seq ◽  
Transcriptomic Response ◽  
Eimeria Maxima ◽  
Ifn Γ ◽  
Production Knowledge ◽  
Immune Related Genes ◽  
Kinetics Of

Eimeria maxima is a common cause of coccidiosis in chickens, a disease that has a huge economic impact on poultry production. Knowledge of immunity to E. maxima and the specific mechanisms that contribute to differing levels of resistance observed between chicken breeds and between congenic lines derived from a single breed of chickens is required. This study aimed to define differences in the kinetics of the immune response of two inbred lines of White Leghorn chickens that exhibit differential resistance (line C.B12) or susceptibility (line 15I) to infection by E. maxima. Line C.B12 and 15I chickens were infected with E. maxima and transcriptome analysis of jejunal tissue was performed at 2, 4, 6 and 8 days post-infection (dpi). RNA-Seq analysis revealed differences in the rapidity and magnitude of cytokine transcription responses post-infection between the two lines. In particular, IFN-γ and IL-10 transcript expression increased in the jejunum earlier in line C.B12 (at 4 dpi) compared to line 15I (at 6 dpi). Line C.B12 chickens exhibited increases of IFNG and IL10 mRNA in the jejunum at 4 dpi, whereas in line 15I transcription was delayed but increased to a greater extent. RT-qPCR and ELISAs confirmed the results of the transcriptomic study. Higher serum IL-10 correlated strongly with higher E. maxima replication in line 15I compared to line C.B12 chickens. Overall, the findings suggest early induction of the IFN-γ and IL-10 responses, as well as immune-related genes including IL21 at 4 dpi identified by RNA-Seq, may be key to resistance to E. maxima.

scholarly journals Vitamin D Reverses Disruption of Gut Epithelial Barrier Function Caused by Campylobacter jejuni

2021 ◽  
Vol 22 (16) ◽  
pp. 8872
Author(s):  
Fábia D. Lobo de Sá ◽  
Steffen Backert ◽  
Praveen K. Nattramilarasu ◽  
Soraya Mousavi ◽  
Geoffrey I. Sandle ◽  
...  
Keyword(s):  
Vitamin D ◽  
Epithelial Cells ◽  
Campylobacter Jejuni ◽  
Epithelial Barrier ◽  
Intestinal Epithelial ◽  
Rna Seq ◽  
Barrier Dysfunction ◽  
Epithelial Barrier Function ◽  
Bacterial Gastroenteritis ◽  
The Relationship

Infections by the zoonotic foodborne bacterium Campylobacter jejuni (C. jejuni) are among the most frequent causes of bacterial gastroenteritis worldwide. The aim was to evaluate the relationship between epithelial barrier disruption, mucosal immune activation, and vitamin D (VD) treatment during C. jejuni infection, using intestinal epithelial cells and mouse models focused on the interaction of C. jejuni with the VD signaling pathway and VD treatment to improve C. jejuni-induced barrier dysfunction. Our RNA-Seq data from campylobacteriosis patients demonstrate inhibition of VD receptor (VDR) downstream targets, consistent with suppression of immune function. Barrier-preserving effects of VD addition were identified in C. jejuni-infected epithelial cells and IL-10−/− mice. Furthermore, interference of C. jejuni with the VDR pathway was shown via VDR/retinoid X receptor (RXR) interaction. Paracellular leakiness of infected epithelia correlated with tight junction (TJ) protein redistribution off the TJ domain and apoptosis induction. Supplementation with VD reversed barrier impairment and prevented inhibition of the VDR pathway, as shown by restoration of transepithelial electrical resistance and fluorescein (332 Da) permeability. We conclude that VD treatment restores gut epithelial barrier functionality and decreases bacterial transmigration and might, therefore, be a promising compound for C. jejuni treatment in humans and animals.

scholarly journals Kinetics of the immune response to Eimeria maxima in relatively resistant and susceptible chicken lines

2019 ◽  
Author(s):  
Abi Bremner ◽  
Sungwon Kim ◽  
Katrina Morris ◽  
Matthew J. Nolan ◽  
Dominika Borowska ◽  
...  
Keyword(s):  
Immune Response ◽  
Post Infection ◽  
Differential Resistance ◽  
Poultry Production ◽  
Rna Seq ◽  
Eimeria Maxima ◽  
Ifn Γ ◽  
Production Knowledge ◽  
Immune Related Genes ◽  
Kinetics Of

AbstractEimeria maxima is a common cause of coccidiosis in chickens, a disease which has a huge economic impact on poultry production. Knowledge of immunity to E. maxima and the specific mechanisms that contribute to differing levels of resistance observed between chicken breeds and between congenic lines derived from a single breed of chickens is required. This study aimed to define differences in the kinetics of the immune response of two inbred lines of White Leghorn chickens that exhibit differential resistance (line C.B12) or susceptibility (line 15I) to infection by E. maxima. Line C.B12 and 15I chickens were infected with E. maxima and transcriptome analysis of infected jejunal tissue was carried out at 2, 4, 6 and 8 days post-infection (dpi). RNA-Seq analysis revealed differences in the rapidity and magnitude of cytokine transcription responses post-infection between the two lines. In particular, IFN-γ and IL-10 transcripts in the jejunum accumulated earlier in line C.B12 (at 4 dpi) compared to line 15I (at 6 dpi). Line C.B12 chickens exhibited increases of IFNG and IL10 mRNA in the jejunum at 4 dpi, whereas in line 15I transcription was delayed but increased to a greater extent. RT-qPCR and ELISAs confirmed the results of the transcriptomic study. Higher serum IL-10 correlated strongly with higher E. maxima replication in line 15I compared to line C.B12 chickens. Overall, the findings suggest early induction of the IFN-γ and IL-10 responses, as well as immune-related genes at 4 dpi identified by RNA-Seq, may be key to resistance to E. maxima.

scholarly journals Evaluation of Glycosylated FlpA and SodB as Subunit Vaccines Against Campylobacter jejuni Colonisation in Chickens

Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 520
Author(s):  
Prerna Vohra ◽  
Cosmin Chintoan-Uta ◽  
Vanessa S. Terra ◽  
Abi Bremner ◽  
Jon Cuccui ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Poultry Meat ◽  
Challenge Dose ◽  
Source Of Infection ◽  
Protein Carriers ◽  
Glycosylation Sites ◽  
Colony Forming Units ◽  
Vaccine Antigens ◽  
Strain Vaccine ◽  
Glycoconjugate Vaccines

Campylobacter jejuni is the leading bacterial cause of human gastroenteritis worldwide and the handling or consumption of contaminated poultry meat is the key source of infection. C. jejuni proteins FlpA and SodB and glycoconjugates containing the C. jejuni N-glycan have been separately reported to be partially protective vaccines in chickens. In this study, two novel glycoproteins generated by protein glycan coupling technology—G-FlpA and G-SodB (with two and three N-glycosylation sites, respectively)—were evaluated for efficacy against intestinal colonisation of chickens by C. jejuni strain M1 relative to their unglycosylated variants. Two independent trials of the same design were performed with either a high challenge dose of 107 colony-forming units (CFU) or a minimum challenge dose of 102 CFU of C. jejuni M1. While antigen-specific serum IgY was detected in both trials, no reduction in caecal colonisation by C. jejuni M1 was observed and glycosylation of vaccine antigens had no effect on the outcome. Our data highlight inconsistencies in the outcome of C. jejuni vaccination trials that may reflect antigen-, challenge strain-, vaccine administration-, adjuvant- and chicken line-specific differences from previously published studies. Refinement of glycoconjugate vaccines by increasing glycosylation levels or using highly immunogenic protein carriers could improve their efficacy.

scholarly journals Comparison of Genotypes and Serotypes of Campylobacter jejuni Isolated from Danish Wild Mammals and Birds and from Broiler Flocks and Humans

2001 ◽  
Vol 67 (7) ◽  
pp. 3115-3121 ◽  
Author(s):  
L. Petersen ◽  
E. M. Nielsen ◽  
J. Engberg ◽  
S. L. W. On ◽  
H. H. Dietz
Keyword(s):  
Campylobacter Jejuni ◽  
Developed Countries ◽  
Human Infection ◽  
Genomic Diversity ◽  
Poultry Meat ◽  
Wild Mammals ◽  
Serotype O ◽  
Source Of Infection ◽  
Strain Collection ◽  
Clonal Lines

ABSTRACT The incidence of human infection with Campylobacter jejuni is increasing in most developed countries and the reason for this is largely unknown. Although poultry meat is considered to be a major source, it is evident that other reservoirs exist, possibly common to humans and poultry. Environmental sources are believed to be important reservoirs of Campylobacter infection in broiler chicken flocks. We investigated the potential importance of wildlife as a source of infection in commercial poultry flocks and in humans by comparing the serotype distributions, fla types, and macrorestriction profiles (MRPs) of C. jejuni isolates from different sources. The serotype distribution in wildlife was significantly different from the known distributions in broilers and humans. Considerable sero- and genotype diversity was found within the wildlife collection, although two major groups of isolates within serotype O:12 and the O:4 complex were found. Common clonal lines among wildlife, chicken, and/or human isolates were identified within serotype O:2 and the O:4 complex. However, MRPs of O:12 and O:38 strains isolated from wildlife and other sources indicated that some clonal lines propagated in a wide selection of animal species but were not detected in humans or broilers in this study. The applied typing methods successfully identified different clonal groups within a strain collection showing large genomic diversity. However, the relatively low number of wildlife strains with an inferred clonal relationship to human and chicken strains suggests that the importance of wildlife as a reservoir of infection is limited.

scholarly journals A novel high-content screening approach for the elucidation of C. jejuni biofilm composition and integrity

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Matthew V. X. Whelan ◽  
Jeremy C. Simpson ◽  
Tadhg Ó Cróinín
Keyword(s):  
Atmospheric Oxygen ◽  
Chicken Meat ◽  
Live Cells ◽  
High Throughput Analysis ◽  
Aerobic Conditions ◽  
Poultry Products ◽  
Source Of Infection ◽  
Fluorescent Markers ◽  
Oxygen Conditions ◽  
Bacterial Gastroenteritis

Abstract Background Campylobacter jejuni is the leading cause of bacterial gastroenteritis worldwide and the main source of infection is contaminated chicken meat. Although this important human pathogen is an obligate microaerophile, it must survive atmospheric oxygen conditions to allow transmission from contaminated chicken meat to humans. It is becoming increasingly evident that formation of biofilm plays a key role in the survival of this organism for extended periods on poultry products. We have recently demonstrated a novel inducible model for the study of adherent C. jejuni biofilm formation under aerobic conditions. By taking advantage of supercoiling mediated gene regulation, incubation of C. jejuni with subinhibitory concentrations of the Gyrase B inhibitor novobiocin was shown to promote the consistent formation of metabolically active adherent biofilm. Results In this study, we implement this model in conjunction with the fluorescent markers: TAMRA (live cells) and SytoX (dead cells, eDNA) to develop a novel systematic high-content imaging approach and describe how it can be implemented to gain quantifiable information about the integrity and extracellular polymeric substance (EPS) composition of adherent C. jejuni biofilm in aerobic conditions. We show that this produces a model with a consistent, homogenous biofilm that can be induced and used to screen a range of inhibitors of biofilm adherence and matrix formation. Conclusions This model allows for the first time a high throughput analysis of C. jejuni biofilms which will be invaluable in enabling researchers to develop mechanisms to disrupt these biofilms and reduce the viability of these bacteria under aerobic conditions.

scholarly journals Genome-Wide Identification and Characterization of Hsf and Hsp Gene Families and Gene Expression Analysis under Heat Stress in Eggplant (Solanum melongema L.)

Horticulturae ◽  
2021 ◽  
Vol 7 (6) ◽  
pp. 149
Author(s):  
Chao Gong ◽  
Qiangqiang Pang ◽  
Zhiliang Li ◽  
Zhenxing Li ◽  
Riyuan Chen ◽  
...  
Keyword(s):  
Heat Stress ◽  
Gene Families ◽  
High Temperature Stress ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Genome Wide ◽  
Motif Composition ◽  
Hsp Genes ◽  
Identification And Characterization

Under high temperature stress, a large number of proteins in plant cells will be denatured and inactivated. Meanwhile Hsfs and Hsps will be quickly induced to remove denatured proteins, so as to avoid programmed cell death, thus enhancing the thermotolerance of plants. Here, a comprehensive identification and analysis of the Hsf and Hsp gene families in eggplant under heat stress was performed. A total of 24 Hsf-like genes and 117 Hsp-like genes were identified from the eggplant genome using the interolog from Arabidopsis. The gene structure and motif composition of Hsf and Hsp genes were relatively conserved in each subfamily in eggplant. RNA-seq data and qRT-PCR analysis showed that the expressions of most eggplant Hsf and Hsp genes were increased upon exposure to heat stress, especially in thermotolerant line. The comprehensive analysis indicated that different sets of SmHsps genes were involved downstream of particular SmHsfs genes. These results provided a basis for revealing the roles of SmHsps and SmHsp for thermotolerance in eggplant, which may potentially be useful for understanding the thermotolerance mechanism involving SmHsps and SmHsp in eggplant.

High Pressure Inactivation of Escherichia coli, Campylobacter jejuni, and Spoilage Microbiota on Poultry Meat

2012 ◽  
Vol 75 (3) ◽  
pp. 497-503 ◽  
Author(s):  
YANG LIU ◽  
MIRKO BETTI ◽  
MICHAEL G. GÄNZLE
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Campylobacter Jejuni ◽  
Resistant Mutant ◽  
Poultry Meat ◽  
E Coli ◽  
Cell Counts ◽  
Meat Spoilage ◽  
Brochothrix Thermosphacta ◽  
Pressure Resistance

This study evaluated the high pressure inactivation of Campylobacter jejuni, Escherichia coli, and poultry meat spoilage organisms. All treatments were performed in aseptically prepared minced poultry meat. Treatment of 19 strains of C. jejuni at 300 MPa and 30°C revealed a large variation of pressure resistance. The recovery of pressure-induced sublethally injured C. jejuni depended on the availability of iron. The addition of iron content to enumeration media was required for resuscitation of sublethally injured cells. Survival of C. jejuni during storage of refrigerated poultry meat was analyzed in fresh and pressure-treated poultry meat, and in the presence or absence of spoilage microbiota. The presence of spoilage microbiota did not significantly influence the survival of C. jejuni. Pressure treatment at 400 MPa and 40°C reduced cell counts of Brochothrix thermosphacta, Carnobacterium divergens, C. jejuni, and Pseudomonas fluorescens to levels below the detection limit. Cell counts of E. coli AW1.7, however, were reduced by only 3.5 log (CFU/g) and remained stable during subsequent refrigerated storage. The resistance to treatment at 600 MPa and 40°Cof E. coli AW1.7 was compared with Salmonella enterica, Shiga toxin–producing E. coli and nonpathogenic E. coli strains, and Staphylococcus spp. Cell counts of all organisms except E. coli AW 1.7 were reduced by more than 6 log CFU/g. Cell counts of E. coli AW1.7 were reduced by 4.5 log CFU/g only. Moreover, the ability of E. coli AW1.7 to resist pressure was comparable to the pressure-resistant mutant E. coli LMM1030. Our results indicate that preservation of fresh meat requires a combination of high pressure with high temperature (40 to 60°C) or other antimicrobial hurdles.

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