Study On The Effective Compounds Group Of Yeast Culture

Yeast culture(YC),as a potential product,which could substitute the antibiotic has many biological functions in animal production,such as improving growth performance and promoting immunity,Improving gastrointestinal microecological balance,to ensure intestinal health and so on.Above all,it has wide application prospect inlivestock production.Yeast culture is a kind of nutrient-rich and complex micro-ecological fermentation product which is produced by the fermentation technology with specific culture medium.Many years of research generally believe that its nutritional care function is realized through its fermentation metabolites.However,there are a great variety of metabolites in yeast culture.Besides a few known nutrients,most of them are unknown components in the form of various compounds.The main or specific effective components are not accurately described and explained.Therefore,What is the main active ingredient and their action relationship is a question that needs further study and confirmation.Based on the previous relevant yeast culture of metabonomics analysis method and multivariate statistical analysis research study ideas such as principal component analysis method,and using GC-MS technological detection method,a series of researches have been conducted to explore " effective compounds groups" of yeast culture.Which provided the reference for reviewing the essence of yeast culture effective group functions and give a objective,accurate and swift evaluation of the efficacy of yeast culture products.The purpose of this chapter is to establish the composition analysis of yeast culture.A method for the simultaneous determination of metabolites in yeast cultures produced at different fermentation times by simultaneous silanization derivatization and GC/MS was developed.N,O-bis(trimethylsilane)trifluoroacetamide(BSTFA with 1% TMCS)as a derivatization reagent,5 different fermentation time yeast cultures,the culture medium and Diamond VXP were derived,then which were measured by GC-MS.The structure identification of metabolites was determined by NIST11 database.The results showed that a total of 85 metabolites were identified.The results of principal component analysis(PCA)showed that the yeast culture samples were significantly separated and showed a dynamic trend.The main components of differentiated yeast cultures were amino acids,organic acids,esters,alcohols,oligosaccharides and so on.This method is suitable for the study of the metabolomics of yeast culture,which creates a different alternative space for the exploration of the yeast culture effective group,and also provides the material research basis for the analysis of its effective group.Through the analysis of the growth performance,immune function,nutrient utilization,cecum microorganism and metabolism of broiler,the best yeast culture used in broiler was optimized,which provided the basis for screening the yeast culture effective group.336 AA broilers(age 1 d)with the similar relative quality were randomly assigned to 7 groups with 6 replicates.Each replicate has 8 broilers.The broilers in the control group(group A)were only fed a basal diet.Treatment groups supplemented with different fermentation time of yeast culture at 12 h,24 h,36 h,48 h and 60 h were named group B,group C,group D,group E,group F,respectively.Treatment G was supplemented with Diamond V XP.The experimental period has 7 d of accilimation and 42 d of growth.The results showed that yeast culture of 24 h fermentative that average daily weight gain(ADG)significantly increased(P<0.05)and the ratio of feed weight(F/G)(P<0.05)was decreased.Moreover,there was a tendancy of increase in average daily feed intake(ADFI)among treatment groups which were increased(P>0.05).At 21 d and 42 d,the IgA and IgG concentrations were significantly higher in cultures of 24 h and 36 h of Saccharomyces cerevisiae(P < 0.05).The concentrations of serum albumin(ABL)and serum total protein(TP)were significantly higher(P < 0.05)in group C than control group and positive control group.Compared with control group,the immune organ indexes of each treatment group have the increased with different degrees.At 42 d,the spleen index in group C was significantly increased by 36.19%(P < 0.05)compared with the control group.In conclusion,under the conditions of this experiment,the nutritional and health efficacy of 24 h fermentative yeast culture on broiler was the best.In addition,The experiment used Illumina Miseq high-throughput sequencing technology to detect the cecal contents of 42-day-old broiler chickens and GC-MS metabolomics to analyze the changes of cecal metabolites in broilers.The results showed that supplementation of yeast cultures increased the relative abundance of intestinal flora in broilers.Among the broilers in group D,the species diversity of the caecal flora was the highest;at the Phylum level,the relative abundance of Firmicutes in each group was the highest,whereas the content of Verrucomicrobia and Akkermansia "slimming bacteria" in fermented 24 h of yeast cultures were significantly higher than other group.The cecal contents of 7 treatment groups were determined by GC-MS.The structure of the compounds was determined by NIST11 database and reference identification.The data were pre-processed and imported into SIMCA-P 13.0 software for multivariate statistical analysis.Results found that the PCA scores showed at individual groups were clustered together at each time point,which means that the addition of yeast culture changed the metabolites.Comparison of yeast cultures of fermented 24 h and 60 h with large difference in production performance,the distinguishing metabolites were screened out by OPLS-DA modeling: alanine,leucine,Phenylacetate,furomannose,arabitol,cholesterol.In the product of group C,the contents of alanine,leucine,and phenylacetic acid were increased;whereas,the contents of furan mannose,arabitol,and cholesterol were decreased.These differential metabolites mainly belong to the metabolic pathways as following: phenylalanine metabolism,alanine metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,proline,leucine and isoleucine metabolism,cholesterol metabolism,steroid hormone synthesis,digestion and absorption of fat;vitamin digestion and absorption.Under the conditions of this test,according to the production performance and the screening methods of biomarker biomarkers,such as OPLS-DA,the biomarkers of yeast culture are selected as glycine,fructose,inositol,Piran galactose and sucrose.At the same time,the functional relationship between the production performance and the yeast culture effective group was established,which further revealed the relationship between the combination of the effective group and the production performance,and provided the reference for the confirmation of the yeast culture effective group.In the trial,a single-factor complete randomized grouping design was used.336 AA broilers with a similar healthy body weight were selected from 1 day old males and females.Male and female were randomly divided into 7 groups,each with 6 replicates and 8 replicates.The test period was 42 days.The control group was fed the basal diet,yeast culture effective group(A,B,C)which with 3 different dosage combinations were fed supplemented on the base diet.The group Dwas fed with the yeast culture fermented for 24 hours and the group E?group F were XP group,Add 0.1% and 1% "XP" feed additives on the basic diets.The results showed that: Compared with the control group,in the production performance,compared with the control group,each added treatment group can significantly increase the daily weight gain of broilers,significantly reduce the feed-to-weight ratio(P<0.05),and also have a daily feed intake.Significantly affected(P < 0.05).The daily weight gain of the compatibility group A?group B and group C were greater than that of the D group(YC24h)(P>0.05),and it was 1.71%,4.96%,and 2.69% higher than that of the D group(YC24h).The feed-to-weight ratio of each treatment group was significantly lower than that of the control group(P<0.05),and the feed-to-weight ratio of the compatibility group C was the lowest.At 7-42 days of age,the daily gain of each group was significantly higher than that of the control group(P<0.05),and the feed intake and feed-weight ratio were significantly lower than those of the control group(P<0.05).However,there was no significant difference between the groups(P>0.05).The weight ratio of group C was the lowest among all treatment groups,which was basically the same as that of Group D(YC24h).At 21 days of age,serum total protein,albumin,and serum globulin levels in the experimental groups were all increased compared with the control group,but the difference was not significant(P<0.05).For Aspartate transaminase,the group A and group B were significantly lower than that in the control group(P<0.05).For alkaline phosphatase,the content in each treatment group was higher than that in the control group,and the content of C in the compatibility group was significantly higher than that in the control group and XP group(P <0.05).For serum urea nitrogen,the content in the compatibility group was significantly lower than that in the control group(P<0.05).At 42 days of age,serum total protein,aspartate aminotransferase,alkaline phosphatase and serum globulin were increased in the experimental group compared with the control group,but there was no significant difference compared with the control group(P>0.05).For serum urea nitrogen,the content of C,E,and F in the compatibility group was significantly lower than that in the control group(P<0.05),with E in the lowest and C in the second.There was no significant difference between group(A?B?C)and D group(XP)in the compatibility group(P>0.05).In the immune function of broilers: At the age of 42 days,the thymus index of each treatment group increased compared with the control group,but it was not significant(P>0.05).For IgA,IgG: 21 days of age and 42 days of age,compared with the control group,the IgA and IgG content of each test treatment group increased.At 21 days of age,the IgG content of the compatibility group C was significantly higher than that of the control group(P<0.05).At 42 days of age,the contents of IgA and IgG in the compatibility group B were significantly higher than those in the control group(P<0.05).For 21-day-old broilers,compared with the control group,the content of MDA in the serum of the compatibility group B and group D was lower than that of the control group and the group XP(group E?F),and the MDA content in the serum of the compatibility group B was lower.The lowest was 62.27% lower than the control group(P>0.05).For 42-day-old broilers,compared with the control group,the serum GPX,SOD and CAT activities in the treated groups increased to varying degrees,while the serum MDA levels increased slightly compared with the control group,but the effect was not.Obvious(P>0.05).Effects on slaughter performance of broilers: Compared with the control group,each treatment group had no significant effect on the slaughter rate,slaughter rate,breast muscle rate,and leg muscle ratio of the 42 d broilers(P>0.05),but there was a certain increase trend.This experiment mainly examined the changes in structure and composition of cecal flora in early broilers.The results showed that the effects of different tests on the structure of intestinal flora in broiler chickens were greater.The addition of yeast cultures in the pre-feeding period significantly changed the structure of cecal microflora in the broilers,but the change in the bacterial community structure was not significant between the effective groups and the YC 24 h group.In summary,compared with the fermentation at 24 h of yeast cultures on performance,immune function,antioxidant capacity and slaughter performance,the effective groups can reach the same level of application.Therefore initially confirmation of glycine,fructose,myo-inositol,galactose,and sucrose,which are the main functional components of the effective compounds group.