Effects Of Diet With The Unsaturated Fatty Acids / Saturated Fatty Acids Ratio Simulating To Sow Milk On Intestinal Functions,intestinal Microbiota Of Weaned Piglets

The lipid content in milk is the important energy source for the growth of young individuals,accounting for 40-55%of the total energy intake.Lipids have been incorporated in commercial diets as the most valuable method to achieve high energetic concentrations to meet pig nutritional requirements.Recently,research has shown that fatty acids,exert beneficial effects on inflammatory bowel diseases,moderate the immune response,improve the intestinal morphology and gut microbial ecosystem,while less studies focused attention on the lipid composition whether has effects on the growth,hepatic lipid metabolism and attenuating inflammation of piglets.In the present study,eighteen 21 days old weaning piglets（Duroc×Landrace×Yorkshire,Body weight（BW）:5.446±0.079 kg）were obtained from Hunan Zhenghong Science and Technology development Co.,Ltd.The piglets were randomly divided into three groups（n=6）:（1）Control group（CON）:Piglets were fed with a soybean（2%）basal diet;（2）Sow milk simulated group（SMS）:Piglets were fed diet with 0.64%soybean oil,1.20%palm oil and 0.16%coconut oil,and UFAs/SFAs stimulate to sow milk composition;（3）Emulsification group（EMS）:Piglets were fed diet with emulsified lipids（0.64%soybean oil,1.20%palm oil and 0.16%coconut oil）.The growth performance and organ indexes,serous biochemical parameters,Serum Free amino acids,liver lipid metabolism,Serum and liver antioxidant index,intestinal histomorphology and functions,intestinal microbiota composition of piglets were determined.The test results show that:1)Effects of diet with UFAs/SFAs simulate to sow milk on the growth performance and organ indexes of weaning piglets.Compared with the control group,there was no difference on the ADG,ADFI and organ indexes of liver,spleen,kidney.But diet with UFAs/SFAs simulate to sow milk had a tendency to increase the ratio of F/G.2)Effects of diet with UFAs/SFAs simulate to sow milk on the serum biochemical parameters and Serum free amino acids metabolism.Compared with the control group,the bio-activity of AST（p=0.027）and the concentration of TP（p=0.037）were decreased by diet with UFAs/SFAs simulate to sow milk,but had a tendency to increase the concentration of CHOL（p=0.070）.Compared with the diet with UFAs/SFAs simulate to sow milk,Emulsification significantly increased the content of GSH,but there was no difference on the Serum free amino acids index.3)Effects of diet with UFAs/SFAs simulate to sow milk on the liver lipid and oxidation reaction Compared with the control group,Emulsification significantly increased the expression of FABP m RNA（p=0.001）,and had a tendency to increase the expression of TGH m RNA（p=0.059）and ACC m RNA（p=0.052）,while no significant difference was found on the CAT,GPX1,SOD₂ of liver.4)Effects of diet with UFAs/SFAs simulate to sow milk on the intestinal barrier.（1）tight junction proteins.Diet with UFAs/SFAs simulate to sow milk increased the expression of occludin in duodenum（p=0.007）and ileum（p=0.008）,but emulsification weakened this affect.Diet with UFAs/SFAs simulate to sow milk decreased the expression of claudin-1 m RNA in jejunum（p=0.001）,but increased in ileum（p=0.047）,while emulsification had an adverse effect in jejunum and ileum.Diet with UFAs/SFAs simulate to sow milk increased the expression of zonula occludens-1（ZO-1）m RNA in duodenum（p<0.001）,and jejunum（p=0.002）,while emulsification only had the same effect in duodenum.（2）pro-inflammatory genes.Diet with UFAs/SFAs simulate to sow milk increased the expression of toll-like receptor 4（TLR4）m RNA（p=0.019）in jejunum,but decreased the expression of TLR4（p=0.041）m RNA in ileum.Emulsification changed the expression of nuclear factor kappa B（NF-κb）,and increased its m RNA expression in duodenum（p<0.001）,while decreased its m RNA expression in jejunum（p=0.015）.（3）About the expression of anti-inflammatory genes,diet with UFAs/SFAs simulate to sow milk pronounced increased the expression of interleukin（IL）-10（p=0.036）m RNA in jejunum and IL-22（p=0.009）m RNA in ileum.Emulsification changed increased the expression of IL-22 m RNA in duodenum（p=0.035）and jejunum（p<0.001）,as well as increased the expression of IL-17 m RNA in duodenum（p<0.001）and jejunum（p<0.001）.5)Effects of diet with UFAs/SFAs simulate to sow milk on the short chain fatty acids concentrations.Compared with the control group,Diet with UFAs/SFAs simulate to sow milk and Emulsification decreased the concentrations of butyric acid（p=0.038）in cecum contents.6)Effects of diet with UFAs/SFAs simulate to sow milk on the intestinal microbiota composition.（1）The operational taxonomic units（OTU）distributions in jejunum,ileum,cecum and colon among the three group were analyzed.The resulted shows that 761,526,627,683 of the total taxa were shared of which 91,80,225;77,153,166;115,60,141and 94,76,136 unique elements among the groups in the jejunum,ileum,cecum,colon,respectively,the unique OTUs number of diets of CON group was much higher than other groups.β-diversity analysis represents the extent of similarity between different microbial communities The Principal Coordinates Analysis（PCo A）using weighted UNIFRAC analysis.Emulsification decreased separation in jejunum（p=0.036）and ileum（p<0.001）.In the colon,Emulsification has a tendency to increase the separation（p=0.056）.（2）The bacterial sequences on the jejunum,ileum,caecum and colon among three groups were analyzed by Nonparametric Test at phylum and genus levels.In the about identified phylum,Emulsification increased abundance of Bacteroidetes in ileum（p=0.027）,Diet with UFAs/SFAs simulate to sow milk increased the abundance of Bacteroidetes in cecum（p=0.002）.In the about identified genus,Emulsification increased the abundance of Prevotella＿1 in ileum and cecum.In ileum,Compared with the control group,Emulsification increased the abundance of Prevotella＿1（p=0.033）,Prevotella＿9（p=0.032）,Prevotellaceae＿NK3B31＿group（p=0.031）,Prevotellacea-UGG-003（p=0.031）,Lachnospiraceae-UGG-005（p=0.031）,Ruminococcaceae＿UCG＿005（p=0.042）.In cecum,compared with the control group,Diet with UFAs/SFAs simulate to sow milk decreased the abundance of Clostridium＿sensu＿stricto＿1（p=0.009）,streptococcus（p=0.005）;Emulsification decreased the abundance of Clostridium＿sensu＿stricto＿1（p=0.022）,streptococcus（p=0.004）.7)The biofunction of microbiota in different intestinal segments of weaning piglet were predicted via PICRUSt analysis.In Cecum,Diet with UFAs/SFAs simulate to sow milk major in Amino Acid（p=0.001）,Energy metabolism（p=0.011）,Metabolism of Cofactors and Vitamins（p=0.002）,Nucleotide Metabolism（p=0.007）,Glycan Biosynthesis and Metabolism（p=0.002）,Metabolism of Terpenoids and Polyketides（p=0.001）,Metabolism of Other Amino Acids（p=0.018）and Biosynthesis of Other Secondary Metabolites（p=0.011）at KEGG pathway level 2.At the same time,Emulsification major in Amino Acid（p=0.003）,Energy metabolism（p=0.025）,Metabolism of Cofactors and Vitamins（p=0.017）,Nucleotide Metabolism（p=0.023）,Metabolism（p=0.043）,Glycan Biosynthesis and Metabolism（p=0.004）,Metabolism of Terpenoids and Polyketides（p=0.017）,Metabolism of Other Amino Acids（p=0.003）and Biosynthesis of Other Secondary Metabolites（p=0.002）.8)Correlations coefficient between intestinal microbiota and tight junction proteins,pro-inflammatory and anti-inflammatory related genes expression and short chain fatty acids concentrations.The relationship between the gut microbiota（the relative abundance of 20 at the genus level）and intestinal related genes expression and short chain fatty acids concentrations was performed by the Spearman’s rank correlation.In jejunum,Prevotella,Bacteroidetes,Faecalibacterium,Alloprevotella showed a significant negative correlation with IL-10,IL-17,IL-22 and Claudin-1,while exhibited a positive correlation with NF-kb.In ileum,Prevotella exhibited a significant positive correlation with IL-10,IL-17,IL-22 and Claudin-1;In cecum,Streptococus,Clostridium＿sensu＿stricto＿1,Lachnospiraceae-UGG-005,Lactobacillus showed a significant positive correlation with butyrate,valerate,but Prevotella exhibited a negative correlation with butyrate and valerate.Meanwhile,in colon,Prevotella also exhibited a negative correlation with butyrate and valerate.In this paper,we through the research route of“growth performance-serum related indexes-liver lipid related indexes-intestinal barrier-intestinal microbiota composition find that diet with UFAs/SFAs simulate to sow milk has a tendency to increase the ratio of F/G,improved the liver lipid index,moderated the intestinal barrier function,immune function,and intestinal microbiota composition,which could affect the intestinal health in weaned piglets.