| The aim of this study was to determine the effect of complex probiotics and it s cultures on microflora,rumen fermentation system and immune function of sheep under different concentrate to forage ratios and detected by real-time quantitative P CR(RT-qPCR).A total of six healthy Mongolian sheep fitted with different add materials with good health and similar weight[average body weight was(45.5 ± 1.52)kg]and r umen fistula were randomly divided into two periods.Using in self-control cross de sign trial,the experiment were crossing conducted in batches and lasted for 35 day s,consisted of early period(15-day)and late period(20-day).In period one,the s heep were divided into two groups,with 3 replicates per group.Sheep in group 1 w ere fed low concentratediet,while sheep in group 2 were fed high-concentrate diet.The diet was exchanged in Period two the after two weeks interval.Each period co mprised three parts of content part I(control group)part II(The sheep were fed probiotics or the cultures ten days),part III(The sheep was fed complex probiotic s or the cultures twenty days).Ruminal content was collected before feeding for t he sample of DNA extract and activity of CMC、Amylase、Protease.Ruminal conte nt was collected before feedingat 2、4、6、8、12 hours to measure Ruminal fermentation Parameter.Finally the Jugular venous blood of sheep were collected bef ore feeding for the sample of sCD4+、sCD8+.The results showed that:(1)The same level of complex probiotics were added under the lower level of concentrate diet.Compared with the control group,the number of General bacteria,F-succinogenes,R-flavefaciens,B-fibrisolvens,Lactobacillus,Ruminobacter amylophilus were significantly increased(P<0.05),whereas the R-albus,Prevotella brevis were significantly decreased(P<0.05);the Anaerobic fungi and S-ruminantiumwere upward trend(P>0.05),and the S-bovis,Methannogens were no change.(2)The same level of complex probiotics cultures were added under the lower level of concentrate diet.Compared with the control group,the number of General bacteria,F-succinogenes were significantly increased(P<0.05).the Anaerobic fungi,R-albus,B-fibrisolvens,S-ruminantium,Lactobacillus,Prevotella-brevis,Methannogens we re significantly decreased(P<0.05).the S-bovis and R-flavefaciens showed a decreased tendency(P>0.05);The difference of Ruminobacter amylophilus were not prominen t(3)The same level of complex probiotics were added under the higher level of concentrate diet.Compared with the control group,the number of Anaerobic.fungi,F-succinogenes,Ruminobacter amylophilus were significantly increased(P<0.05);the S-bovis,Methannogens were significantly decreased(P<0.05);the B-fibrisolvens,S-ruminantium,Lactobacillus,Prevotella brevis showed a increased tendency(P>0.05);whereas the S-bovis,the General bacteria,R-albus,R-flavefaciens were decreased(P>0.05);(4)The same level of complex probiotics cultures were added under the higher level of concentrate diet.Compared with the control group,the number of Anaerobic fungi,S-bovis,Ruminobacter amylophilus were significantly increased(P<0.05);the General bacteria,F-succinogenes,R-albus,R-flavefaciens,B-fibrisolvens,S-ruminantium were significantly decreased(P<0.05);the Lactobacillus showed a decreased tendency(P>0.05);the Prevotella brevis,Methannogens was no change.(5)The same level of complex probiotics and the cultures were added under the different level of concentrate diet.Compared with the control group,the pH was increased,and the concentration of NH3-N wered decreased As well as the increased the TVFA.The complex probiotics addition werebetter than its cultures.(6)The same level of complex probiotics and the cultures were added under the different level of concentrate diet.Compared with the control group,the activity of Protease was significantly increasing(P<0.05);the activity of CMC was significantly decreasing under the lower level of concentrate diet and significantly increasing under the lower level of concentrate(P<0.05).The Amylase stay a stable stage.(7)The same level of complex probiotics and the cultures were added under the different level of concentrate diet.Compared with the control group,the concentration of sCD4+was significantly increased under the same level of complex probiotics addition(P<0.05),but the concentration of sCD8+ showed decreased trendency under the same cultures.However,The the concentration of sCD8+ was significantly increased under the lower level of concentrate(P<0.05. |
