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Swine intestinal microbiota perturbation by antibiotic treatment is linked to a reduced number of necrotic epithelial cells and up-regulation of immune-associated pathways using an in vitro Salmonella Typhimurium challenge model

ABSTRACT: Salmonella enterica serovar Typhimurium is an animal welfare and public health concern due to its ability to parasite livestock and potentially contaminate meat products. In pigs, it is associated with enterocolitis and diarrhea, whereas some individuals may become life-long carriers. To reduce Salmonella shedding and the risk of pork contamination, antibiotic therapy is used and can contribute to antimicrobial resistance. Here we hypothesized that immune system education by the microbiota can play a role in intestinal resilience to infection. We used amoxicillin (15mg/Kg) to modulate the intestinal microbiome of 10 piglets, paired with same age pigs that received a placebo (n=10) from 0 to 14 days of age. Animals were euthanized when 4-weeks old. Each pig donated colon sections for ex vivo culture (n=20 explants/pig). Explants were inoculated with S. Typhimurium, PBS or LPS. The gut bacteriome was characterized by sequencing of the 16S rRNA at 7, 21 days of age, and upon in vitro culture. In vivo antibiotic treatment led to b-diversity differences between groups at all times (P<0.05), while a-diversity did not differ between amoxicillin and placebo groups on day 21 and at euthanasia (P<0.03 on day 7). Explant microbiomes were not different from in vivo. No significant differences in epithelial cell necrosis scores were observed between amoxicillin and placebo groups after exposure to Salmonella, even though median scores were lower in the amoxicillin group. Both groups did have significantly higher necrosis scores than the PBS group (P<0.05). Activation of immune-related cascades in control explants from the amoxicillin group was observed, whereas Salmonella exposure activated specific cellular repair, wound healing and macrophage pathogen inhibition pathways that were significantly different from placebo pigs. These suggest that immune education by the microbiota may be developed as a tool to mitigate intestinal lesions following pathogen exposure.

ORGANISM(S): Sus scrofa

PROVIDER: GSE134130 | GEO | 2020/04/08

REPOSITORIES: GEO

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Project description:A first generation Affymetrix GeneChipÂ® Porcine genome array was used to profile the gene expression in porcine mesenteric lymph nodes over a time course of infection with S. Typhimurium, including the acute (8 hours post inoculation (hpi), 24 hpi, 48 hpi) and chronic (21 days post-inoculation (dpi)) stages of infection. Our objectives were to 1) identify and examine the stereotypical gene expression response within host MLN to S. Typhimurium infection, 2) characterize global host responses by revealing the specific features of the hostâs innate immunity pathways, and 3) explore if and how S. Typhimurium may escape the host immune response and develop into a carrier state. Our study has attempted to investigate the features of host gene expression profiling during S. Typhimurium infection at the acute and chronic infection stages and to explore the mechanism by which S. Typhimurium can escape from the host immune response and develop a carrier state in the host. In conclusion, by using the Affymetrix porcine GeneChip, 848 differentially expressed genes were identified in porcine MLN during infection and several specific features of host response were revealed by gene cluster and pathway analysis. Our data are the first report to investigate global host responses to S. Typhimurium in porcine MLN, and this new study provides data applicable for studying enteric salmonellosis of pigs, as well as humans. Experiment Overall Design: Fifteen piglets from Salmonella spp.-free sows were weaned at 10 days (d) of age, shipped to the National Animal Disease Center, Ames, IA and raised in isolation facilities. To confirm that all piglets were free of Salmonella spp. prior to challenge, bacteriological cultures were performed on rectal swabs twice. Seven week old pigs were randomly divided into 2 groups, 3 non-infected pigs and 12 infected pigs. Three non-infected control pigs were necropsied 2 days prior to experimental infection. On day 0, pigs in the infected groups were intranasally challenged with 1 billion CFU of Salmonella enterica serovar Typhimurium. Three infected pigs were necropsied at 8 hours post-inoculation (hpi), 24 hpi, 48 hpi and 21 days post-inoculation (dpi). Tissue samples from the mesenteric lymph nodes (MLN) were collected and immediately frozen in liquid nitrogen for RNA isolation.

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Swine intestinal microbiota perturbation by antibiotic treatment is linked to a reduced number of necrotic epithelial cells and up-regulation of immune-associated pathways using an in vitro Salmonella Typhimurium challenge model

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