Low-protein diets supplemented with casein hydrolysate favor the microbiota and enhance the mucosal humoral immunity in the colon of pigs

Background:High-protein diets can increase the colonic health risks.A moderate reduction of dietary crude-protein(CP)level can improve the colonic bacterial community and mucosal immunity of pigs.However,greatly reducing the dietary CP level,even supplemented with all amino acids(AAs),detrimentally affects the colonic health,which may be due to the lack of protein-derived peptides.Therefore,this study evaluated the effects of supplementation of casein hydrolysate(peptide source)in low-protein(LP)diets,in comparison with AAs supplementation,on the colonic microbiota,microbial metabolites and mucosal immunity in pigs,aiming to determine whether a supplementation of casein hydrolysate can improve colonic health under very LP level.Twenty-one pigs(initial BW 19.90±1.00 kg,63±1 days of age)were assigned to three groups and fed with control diet(16%CP),LP diets(13%CP)supplemented with free AAs(LPA)or casein hydrolysate(LPC)for 4 weeks.Results:Compared with control diet,LPA and LPC diet decreased the relative abundance of Streptococcus and Escherichia coli,and LPC diet further decreased the relative abundance of Proteobacteria.LPC diet also increased the relative abundance of Lactobacillus reuteri.Both LP diets decreased concentrations of ammonia and cadaverine,and LPC diet also reduced concentrations of putrescine,phenol and indole.Moreover,LPC diet increased total short-chain fatty acid concentration.In comparison with control diet,both LP diets decreased protein expressions of Toll-like receptor-4,nuclear factor-κB,interleukin-1βand tumor necrosis factor-α,and LPC diet further decreased protein expressions of nucleotide-binding oligomerization domain protein-1 and interferon-γ.LPC diet also increased protein expressions of G-protein coupled receptor-43,interleukin-4,transforming growth factor-β,immunoglobulin A and mucin-4,which are indicators for mucosal defense activity.Conclusions:The results showed that supplementing casein hydrolysate showed beneficial effects on the colonic microbiota and mucosal immunity and barrier function in comparison with supplementing free AAs in LP diets.These findings may provide new framework for future nutritional interventions for colon health in pigs.

Low crude protein diets supplemented with casein hydrolysate enhance the intestinal barrier function and decrease the pro-inflammatory cytokine expression in the small intestine of pigs

To reduce nitrogen excretion and lower feeding costs,low crude protein(CP)diets are sometimes pro-posed,however,a great reduction of dietary CP concentration(>4%reduction vs.recommended con-centration),even supplemented with essential and nonessential amino acids(AA)can detrimentally affect small intestinal barrier function and immunity,possibly due to the excessive lack of peptides.Here we hypothesize that with an extremely low CP concentration diet,protein-derived peptides,rather than AA supplementation,can improve intestinal barrier development and health.To test this hypothesis,21 growing pigs(19.90±1.00 kg body weight)were randomly assigned to 3 treatments with control diet(16%CP),or low CP diets(13%CP)supplemented with AA(LCPA)or casein hydrolysate(LCPC)for 28 days.In comparison with the control diet,the LCPA diet decreased the protein expression level of jejunal barrier factor zonula occludens-1(ZO-1)and stem cell proliferation factor leucine-rich repeat-containing G-protein-coupled receptor-5,whereas the LCPC diet enhanced intestinal barrier function by increasing the protein expression level of jejunal occludin and ZO-1 and ileal mucin-2.The LCPA diet reduced Lactobacillus counts,whereas the LCPC diet increased Lactobacillus counts and reduced Escherichia coli counts in the ileum.The LCPA diet also increased protein expression levels of pro-inflammatory cytokine interleukin-6(IL-6)and IL-22,whereas the LCPC diet decreased protein expression levels of pro-inflammatory IL-1β,IL-17A and tumor necrosis factor-αin the ileum.Collectively,the casein hydroly-sate supplementation of low CP diets showed beneficial effects on the small intestinal barrier,bacterial community,and immunity in pigs,pointing to the important role of protein-derived peptides in small intestinal health in cases of low crude protein diets.