Genome analysis of C. jejuni and C. coli isolated from different regions of the same chicken
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ABSTRACT:
ORGANISM(S): Campylobacter jejuni subsp. jejuni NCTC 11168 = ATCC 700819 Campylobacter coli Campylobacter jejuni RM1221 Campylobacter coli RM2228 Campylobacter jejuni
PROVIDER: GSE17805 | GEO | 2015/09/15
SECONDARY ACCESSION(S): PRJNA118321
REPOSITORIES: GEO
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Project description:The gut of chicken is mostly colonised with Campylobacter jejuni and with 100 fold less C. coli. The competitive ability of C. coli OR12 over C. jejuni OR1 has been examined in experimental broiler chickens following the observation that C. coli replaced an established C. jejuni intestinal colonisation within commercial chicken flocks reared outdoors (El-Shibiny, A., Connerton, P.L., Connerton, I.F., 2005. Enumeration and diversity of campylobacters and bacteriophages isolated during the rearing cycles of free-range and organic chickens. Applied Environmental Microbiology. 71, 1259-1266). Five independent DNA preps of C. jejuni RM1221 were labelled with Cy 5 independently and they were mixed well which was used as the control. OR1 and OR12 were labelled with Cy 3 independently and equal concentration of the control and sample DNA were used for hybridisation. Three biological replicates were done for each slide. The supplementary file (linked at the foot of this record) represents the averaged normalised values for each experimental condition (3replicates/experimental condition).
2015-09-15 | E-GEOD-17805 | biostudies-arrayexpress
Project description:C. jejuni HPC5 is a Campylobacter strain isolated from chickens. Following bacteriophage CP34 treatment on chickens colonised by C. jejuni HPC5, a series of CP34 insensitive strains like C. jejuni HPC5 R14 and C. jejuni HPC5 R20 were obtained which compromised their ability to colonise chickens. Reintroduction of C. jejuni HPC5 R14 and R20 in to chickens led to reversion of these strains and the MRPs of the revertant strains fell in to different classes termed C. jejuni HPC5 R14A, R14B, R20A, R20B and R20C and these strained were tested positive for colonisation proficient and bacteriophage sensitive.
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Project description:C. jejuni HPC5 is a Campylobacter strain isolated from chickens. Following bacteriophage CP34 treatment on chickens colonised by C. jejuni HPC5, a series of CP34 insensitive strains like C. jejuni HPC5 R14 and C. jejuni HPC5 R20 were obtained which compromised their ability to colonise chickens. Reintroduction of C. jejuni HPC5 R14 and R20 in to chickens led to reversion of these strains and the MRPs of the revertant strains fell in to different classes termed C. jejuni HPC5 R14A, R14B, R20A, R20B and R20C and these strained were tested positive for colonisation proficient and bacteriophage sensitive. There are three biological replicates for each experiment. Seven independent cDNA preps of C. jejuni HPC5 were prepared and they were labelled independently using AF555. The labelled cDNAs were mixed together and this acted as the control. Each daughter strains were considered as sample and they were labelled with AF645. Hybridisations were done between C. jejuni HPC5 and C. jejuni HPC5 as one of the sample and each of the daughter strains in triplicates. The supplementary files (linked at the foot of this record) contain the summary of the replicate data of the corresponding strain (3 replicates/strain) which have been averaged.
2015-09-15 | E-GEOD-17799 | biostudies-arrayexpress
Project description:Avian pathogenic Escherichia coli (APEC) is a subset of extraintestinal pathogenic E. coli that causes detrimental losses to the poultry industry. Vaccines to reduce APEC in chickens have been partially successful, but many lack protection against multiple serotypes of APEC. Recombinant attenuated Salmonella vaccine (RASV) strains have been used to induce immunity against Salmonella in production chickens and can be modified to deliver foreign antigens as well. This study evaluated the transcriptome of chicken spleens and assessed prevention of APEC infection following vaccination with RASV strains, including a RASV carrying an E. coli antigen. Four-day-old White Leghorn chicks were orally vaccinated with RASV c8025(pYA3337) carrying an empty plasmid, c8025(pYA4428) carrying genes for E. coli common pilus (ECP), a combination of RASVs c8025(pYA3337) and c8025(pYA4428), or PBS (unvaccinated). To assess the host response to vaccination, antibody titers were measured by ELISA and spleen samples (n = 5) were collected from combination vaccinated and unvaccinated groups of four-week-old chickens for RNA sequencing. Five-week old chickens were challenged via air sac with APEC strains APEC-O2 and c7122 (O78). Blood was obtained 24 hours post-challenge, heart, liver, lung, and spleen were collected 48 hours post-challenge for enumeration of E. coli, and gross colibacillosis lesions were scored at necropsy. Chickens vaccinated with RASV strains elicited anti-E. coli EcpA, as well as cross reactive anti-E. coli IutA and IroN IgY antibodies. IgA results. In some organs, bacterial loads and lesions scores were numerically reduced, but no significant differences were detected for vaccinated compared to unvaccinated chickens. Transcriptome results. This data indicates that RASVs could be used to stimulate the immune system and is an initial step toward developing improved therapeutics to combat APEC infections in chickens.
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Project description:Campylobacter jejuni (C. jejuni) is a zoonotic pathogen that causes human diarrhea worldwide. Chickens are a natural reservoir of C. jejuni. Understanding the host response to C. jejuni infection at the molecular level will lay the foundation to control human campylobacterosis by reducing food contamination. Two distinct genetic lines, resistant (line A) and susceptible (line B) to C. jejuni colonization, were utilized to profile the host response to C. jejuni infection using an Agilent chicken 44K microarray. Day-old chickens were challenged orally with C. jejuni and spleens collected for total RNA 7 days post-challenge. Twenty infected samples with highest (a) or lowest bacterial number (b) in cecal content and twenty non-infected (c) in each line were randomly pooled into four biological replicates. The pair comparisons among these three groups within each line were analyzed. The signal intensity of each gene was normalized using LOWESS method. A mixed model was used to identify differentially expressed genes by SAS (P < 0.001). This was opposite to previous cecal tonsil microarray result. There were 468, 743, and 939 genes differentially expressed between groups a and c, groups a and b, and groups b and c in line A, respectively, and 201, 37, 126 genes in line B, respectively. More differentially expressed genes in spleen in line A than in line B were found. The results indicated that significantly different response to C. jejuni infection occurred between resistant and susceptible chicken lines, and the effects of interaction between genetics and tissue should be considered.
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Project description:Campylobacter jejuni is a human pathogen which causes campylobacteriosis, one of the most widespread zoonotic enteric diseases worldwide. Most cases of sporadic C. jejuni infection occur through the handling or consumption of undercooked chicken meat, or cross-contamination of other foods with raw poultry fluid. A common practice to combat Campylobacter infection is to treat chickens with chlorine which kills the microbe. This analysis aimed to elucidate the transcriptomic response of Campylobacter jejuni treated with hypochlorite through Illumina sequencing. C. jejuni was grown and treated with hypochlorite. Samples were taken 5, 20 and 45 min after treatment for RNAseq analysis.The data generated were compared to the transcriptome pre-exposure to determine C. jejuni's response to hypochlorite.
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Project description:Campylobacter jejuni (C. jejuni) is a zoonotic pathogen that causes human diarrhea worldwide. Chickens are a natural reservoir of C. jejuni. Understanding the host response to C. jejuni infection at the molecular level will lay the foundation to control human campylobacterosis by reducing food contamination. Two distinct genetic lines, resistant (line A) and susceptible (line B) to C. jejuni colonization, were utilized to profile the host response to C. jejuni infection using an Agilent chicken 44K microarray. Day-old chickens were challenged orally with C. jejuni and spleens collected for total RNA 7 days post-challenge. Twenty infected samples with highest (a) or lowest bacterial number (b) in cecal content and twenty non-infected (c) in each line were randomly pooled into four biological replicates. The pair comparisons among these three groups within each line were analyzed. The signal intensity of each gene was normalized using LOWESS method. A mixed model was used to identify differentially expressed genes by SAS (P < 0.001). This was opposite to previous cecal tonsil microarray result. There were 468, 743, and 939 genes differentially expressed between groups a and c, groups a and b, and groups b and c in line A, respectively, and 201, 37, 126 genes in line B, respectively. More differentially expressed genes in spleen in line A than in line B were found. The results indicated that significantly different response to C. jejuni infection occurred between resistant and susceptible chicken lines, and the effects of interaction between genetics and tissue should be considered. Chickens in two broiler lines were inoculated with 10^5 cfu C. jejuni on one day after hatch. The cecal content and cecal tonsil was collected and bacterial number in cecal content was counted on day 7 after inoculation. Twenty samples were separated into 3 groups (high burden, low burden, and control) based on bacterial burden of cecal content in each line, 5 samples were mixed randomly into one pool. A dual color, balanced design was carried on for all samples. Three comparisons were used in each line, non-infected/susceptible, susceptible/resistant, resistant/non-infected, totally, four biological replicates in each line. A Dye swap was used in each pair of comparisons including AN/AS, AS/AR, AR/AN; BN/BS, BS/BR, and BR/BN. Background subtracted signal intensity were collected from 24 arrays and normalized for data analysis.
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