Bacillus sp. probiotic supplementation diminish the Escherichia coli F4ac infection in susceptible weaned pigs by influencing the intestinal immune response, intestinal microbiota and blood metabolomics

Background:Probiosis is considered a potential strategy to reduce antibiotics use and prevent postweaning diarrhea(PWD).This study investigated the effect of Bacillus amyloliquefaciens DSM25840 or Bacillus subtilis DSM25841 supplementation on growth,health,immunity,intestinal functionality and microbial profile of post-weaning pigs after enterotoxigenic E.coli(ETEC)F4 challenge.Methods:Sixty-four post-weaning piglets(7748 g±643 g)were randomly al ocated to four groups:control basal diet(CO);CO+1.28×10~6 CFU/g of B.amyloliquefaciens(BAA);CO+1.28×10~6 CFU/g feed of B.subtilis(BAS);CO+1 g colistin/kg of feed(AB).At day(d)7,animals were chal enged with 10~5 CFU/m L of ETEC F4 ac O149 and then fol owed for fecal score and performance until d 21.Blood was col ected at d 6,d 12 and d 21 for immunoglobulins,at d 8 for acute phase proteins,at d 8 and d 21 for metabolomics analysis.Jejunum was sampled for morphometry,quantification of apoptosis,cel proliferation,neutral and acid mucine and Ig A secretory cel s,and microarray analysis at d 21.Jejunum and cecum contents were col ected for microbiota at d 21.Results:AB and BAS reduced the fecal score impairment compared to CO(P<0.05)at d 14.Body weight(BW),average daily weight gain(ADWG),average daily feed intake(ADFI)and gain to feed ratio(G:F)did not differ between Bacil us groups and CO.AB improved BW at d 7,d 14 and d 21,ADWG ADFI and G:F from d 0 to d 7(P<0.05).At d 8,CO had higher plasma arginine,lysine,ornithine,glycine,serine and threonine than other groups,and higher haptoglobin than AB(P<0.05).At d 21,CO had lower blood glycine,glutamine and Ig A than BAS.Morphology,cel s apoptosis and mucins did not differ.BAS and AB increased the vil us mitotic index.Transcriptome profile of BAS and AB were more similar than CO.Gene sets related to adaptive immune response were enriched in BAA,BAS and AB.CO had enriched gene set for nuclear structure and RNA processing.CO had a trend of higher Enterobacteriaceae in cecum than the other groups(P=0.06).Conclusion:Bacil us subtilis DSM25841 treatment may reduce ETEC F4ac infection in weaned piglets,decreasing diarrhea and influencing mucosal transcriptomic profile.

Pharmacological inhibition of cannabinoid receptor 1 stimulates gastric release of nesfatin-1 via the mTOR pathway

AIM To determine whether Nucb2/nesfatin1 production is regulated by the cannabinoid system through the intracellular m TOR pathway in the stomach.METHODS Sprague Dawley rats were treated with vehicle, rimonabant, rapamycin or rapamycin+rimonabant. Gastric tissue obtained from the animals was used for biochemical assays: Nucb2 m RNA measurement by real time PCR, gastric Nucb2/nesfatin protein content by western blot, and gastric explants to obtain gastric secretomes. Nucb2/nesfatin levels were measured in gastric secretomes and plasma using enzyme-linked immunosorbent assay. RESULTS The inhibition of cannabinoid receptor 1(CB1) by the peripheral injection of an inverse agonist, namely rimonabant, decreases food intake and increases the gastric secretion and circulating levels of Nucb2/nesfatin-1. In addition, rimonabant treatment activates m TOR pathway in the stomach as showed by the increase in pm TOR/m TOR expression in gastric tissue obtained from rimonabant treated animals. These effects were confirmed by the use of a CB1 antagonist, AM281. When the intracellular pathway m TOR/S6 k was inactivated by chronic treatment with rapamycin, rimonabant treatment was no longer able to stimulate the gastric secretion of Nucb2/nesfatin-1.CONCLUSION The peripheral cannabinoid system regulates food intake through a mechanism that implies gastric production and release of Nucb2/Nesfatin-1, which is mediated by the m TOR/S6 k pathway.