Dataset Information

Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens.

ABSTRACT: BACKGROUND:Salmonella enterica serovar Pullorum is a host-restricted serotype causing infection in poultry. The pathogen can not only cause acute infection in young chicks with high mortality and morbidity, but also persist in adult chickens without evident clinical symptoms and lead to vertical transmission. To eradicate S. Pullorum in poultry farms, it is necessary to establish an efficient method to monitor the prevalence of the pathogen in adult chickens. The protein IpaJ is a specific immunogen in S. Pullorum and is not detected in closely related serotypes, such as S. Gallinarum and S. Enteritidis. RESULTS:In the present study, IpaJ was expressed as a recombinant fusion protein MBP-IpaJ in E. coli. The purified MBP-IpaJ was used as a coating antigen to develop an indirect ELISA assay, which was applied to the detection of S. Pullorum infection in chickens. The indirect ELISA assay demonstrated that antibodies produced against IpaJ were detectable in antisera of chickens infected with S. Pullorum in the second week, stably increased until the tenth week, and persisted at a high level in the following two weeks. Furthermore, the ELISA method detected four positive samples out of 200 clinical antiserum samples collected from a poultry farm, and the positive samples were confirmed to be reacted with S. Pullorum using the standard plate agglutination test. CONCLUSIONS:The established indirect ELISA using the IpaJ protein is a novel method for specific detection of S. Pullorum infection, and contribute to eradication of pullorum disease in the poultry industry.

SUBMITTER: Li Q

PROVIDER: S-EPMC6318851 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens.

Li Qiuchun Q Zhu Yue Y Yin Kequan K Xu Lijuan L Yin Chao C Li Yang Y Ren Jingwei J Yuan Yu Y Jiao Xinan X

BMC veterinary research 20190103 1

<h4>Background</h4>Salmonella enterica serovar Pullorum is a host-restricted serotype causing infection in poultry. The pathogen can not only cause acute infection in young chicks with high mortality and morbidity, but also persist in adult chickens without evident clinical symptoms and lead to vertical transmission. To eradicate S. Pullorum in poultry farms, it is necessary to establish an efficient method to monitor the prevalence of the pathogen in adult chickens. The protein IpaJ is a specif ...[more]

PMID: 30606183

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Project description:Pullorum disease is one of the most common diarrhea-related diseases caused by Salmonella enterica subspecies enterica serovar Gallinarum biovar Pullorum (S Pullorum); it negatively affects the poultry industry. However, limited studies have explored the association between the gut microbiota and S Pullorum infection in chickens. In the present study, we performed a microbiome comparison and a microbiome genome-wide association study (mGWAS) to investigate the association among the host genetics, the gut microbiota, and pullorum disease in chickens. We found that S Pullorum infection in chickens could alter the abundance of 39 bacterial genera (P < 0.05). The altered structure and composition of the gut microbiota were also detected in the offspring. mGWAS results revealed host genetic variants to be prominently associated with gut microbial diversity and individual microbes. The pathogens Pelomonas and Brevundimonas, which had a high abundance in positive parent chickens and their offspring, were significantly associated with several genetic mutations in immunity-related genes, such as TGIF1, TTLL12, and CCR7 This finding explained why Pelomonas and Brevundimonas were heritable in S Pullorum-infected chickens. The heritable gut microbes and identified genetic variants could provide references for the selection of resistant chickens and the elimination of pullorum disease.IMPORTANCE The present study investigated the association among the host genome, the gut microbiome, and S Pullorum infection in chickens. The results suggested that the gut microbial structure is altered in S Pullorum-infected chickens. The diversity and abundance of the gut microbiota remarkably differed between the offspring coming from S Pullorum-positive and S Pullorum-negative chickens. Heritable gut microbiota were detected in the offspring. Moreover, host genetic variants were associated with microbial diversity and individual gut microbes. The pathogens Pelomonas and Brevundimonas, which exhibited a high heritability in S Pullorum-positive parents and their offspring, were associated with several genetic mutations in immunity-related genes.

Project description:Salmonella enterica subspecies enterica serovar Gallinarum biovar Pullorum (S Pullorum) is the etiological agent of pullorum disease, causing white diarrhea with high mortality in chickens. There are many unsolved issues surrounding the epidemiology of S Pullorum, including its origin and transmission history as well as the discordance between its phenotypic heterogeneity and genetic monomorphism. In this paper, we report the results of whole-genome sequencing of a panel of 97 S Pullorum strains isolated between 1962 and 2014 from four countries across three continents. We utilized 6,795 core genome single nucleotide polymorphisms (SNPs) to reconstruct a phylogenetic tree within a spatiotemporal Bayesian framework, estimating that the most recent common ancestor of S Pullorum emerged in ?914 CE (95% confidence interval [95%CI], 565 to 1273 CE). The extant S Pullorum strains can be divided into four distinct lineages, each of which is significantly associated with geographical distribution. The intercontinental transmissions of lineages III and IV can be traced to the mid-19th century and are probably related to the "Hen Fever" prevalent at that time. Further genomic analysis indicated that the loss or pseudogenization of functional genes involved in metabolism and virulence in S Pullorum has been ongoing since before and after divergence from the ancestor. In contrast, multiple prophages and plasmids have been acquired by S Pullorum, and these have endowed it with new characteristics, especially the multidrug resistance conferred by two large plasmids in lineage I. The results of this study provide insight into the evolution of S Pullorum and prove the efficiency of whole-genome sequencing in epidemiological surveillance of pullorum disease.IMPORTANCE Pullorum disease, an acute poultry septicemia caused by Salmonella Gallinarum biovar Pullorum, is fatal for young chickens and is a heavy burden on poultry industry. The pathogen is rare in most developed countries but still extremely difficult to eliminate in China. Efficient epidemiological surveillance necessitates clarifying the origin of the isolates from different regions and their phylogenic relationships. Genomic epidemiological analysis of 97 S Pullorum strains was carried out to reconstruct the phylogeny and transmission history of S Pullorum. Further analysis demonstrated that functional gene loss and acquisition occurred simultaneously throughout the evolution of S Pullorum, both of which reflected adaptation to the changing environment. The result of our study will be helpful in surveillance and prevention of pullorum disease.

Dataset Information

Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens.

Publications

Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens.

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