Abstract
BackgroundCampylobacter 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.ResultsThree-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.ConclusionsOur 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.
Highlights
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
Challenge of line 61 and line N birds with C. jejuni M1 confirms differential resistance early after inoculation To examine the level of resistance and susceptibility of lines 61 and N to colonisation by C. jejuni M1, threeweek-old birds from each line were challenged with 108 colony forming units (CFU) of C. jejuni M1 and the resulting numbers of C. jejuni in the caecal content determined at 1 and 5 days post-infection
Transcriptional responses to infection in chicken lines differing in C. jejuni resistance To explore transcriptomic differences underlying the relative resistance of line 61 and susceptibility of line N to C. jejuni M1 colonisation, RNA-Seq analysis was performed on caecal tonsil tissue from both infected and age-matched uninfected control birds of both lines
Summary
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. The caeca are a key site of persistence of Campylobacter in poultry, where numbers of C. jejuni can reach as high as 1010 colony forming units (CFU)/g of contents in the absence of overt pathology. Given such levels, contamination of carcasses with numbers of C. jejuni predicted to be adequate for human infection is challenging to prevent during the slaughter process [8]. No commercial vaccines exist and it is likely that a multifactorial approach will be required
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