Isolation And Identification Of Feeding Microorganisms And Application Effect Of Fermented Pleurotus Erngii Feeds

The purpose of this study was to screen for Bacillus,yeast and lactobacillus strains with strong cellulolytic ability,lignin decomposition ability and antibacterial activity,and to optimize the strains for fermenting Pleurotus eryngii substrate?PES?in order to improve its feed quality.Furthermore,we explored the effect of fermented PES feed on growth performance and serum biochemical parameters in Matou goat.Exp.1 Screening of special microorganisms for fermented PES.The bacteria/fungi were isolated from effective microorganisms?EM?solution fermented PES by using the cellulose-Congo red,Azure-B and De Man,Rogosa,Sharpe-Ca?MRS-Ca?screening medium and testing the cellulase activity,ligninase activity and antibacterial activity.Species were identified by bacterial 16S RNA and fungal 18S rDNA gene sequence analysis.Then,microbial strains we selected were mixed with isovolumetric to make a compound microbial inoculants.The results showed that:We obtained 6 strains:Bacillus tequilensis strain P11,Pichia fermentans strain R8 and Kluyveromyces marxianus strain MU5 with higher cellulase activity,Bacillus amyloliquefaciens subsp.plantarum strain MU7 with higher ligninase activity,Weissella paramesenteroides strain R4 and Pediococcus acidilactici strain R9 with higher antibacterial activity.Exp.2 Effect of compound microbial inoculants on PES feed quality.In this experiment,PES,the raw material,mixed with 5%bran,0.3%brown sugar and0.85%salt,was used as the fermenting substrate.The test group was inoculated with 4%composite microbial inoculants,while the control group was added 4%sterile liquid medium.For both groups,the matrix's moisture content was adjusted to 40%and fermented anaerobically at 30?.The pH and lactic acid?LA?content of matrix were measured 48 h apart until they became stable.Then,the nutrients of the fermented matrix were measured and compared with EM solution fermented PES which conducted with same process.The results showed that:After 7-day fermentation,the quality of the nutrients in the test group,is the best.The pH,neutral detergent fiber?NDF?and acidic detergent fiber?ADF?of the test group were significantly lower than those of the control group?P<0.05?,the LA content was not significantly different?P>0.05?between two groups,the aflatoxin B1?AFB1?content as low as 1.92?g·kg^-1.Furthermore,its feed quality was significantly better than the EM solution fermented substrate?P<0.05?.Exp.3 Effect of fermented PES on growth performance and serum biochemical parameters in fattening goats.In this experiment,60 healthy Matou goats?3-month-old,wether?with an average body weight of?20.99±3.90?kg were randomly divided into 5 groups,experimental groups were fed with composite microbial inoculants fermented PES?20%?25%?30%and 35%?while control group was fed with EM solution fermented PES?30%?.After feeding for 47 days,all animals were determined production performance and serum biochemical parameters.Results were as follows:?1?Compared with the control group,the final body weight?FBW?and average daily gain?ADG?of the compound microbial fermentation group were significantly increased?P<0.05?,the feed weight ratio?F/G?was significantly declined?P<0.05?,and there was no significant difference in dry matter intake?DMI?between treatment and control group?P>0.05?.With the increasing of PES content in the diet,the growth performance of the test sheep in each group has an increasing trend.When adding 35%PES,FBW,ADG and DMI reached the highest value and significantly higher than other groups?P<0.05?,but there was no significant difference in F/G between different groups?P>0.05?.?2?Contents of GLU,Ca,TP,ALB,TC,HDL,LDL and CREA,as well as activity of AST,ALT,LDH,CK,CAT,SOD and GSH-Px in compound microbial fermentation group's test goats were significantly higher than the control group?P<0.05?.Content of UREA was significantly lower than the control group?P<0.05?.Contents of serum P,K,Na,Cl,IgA,IgG,IgM,GLB,TG and MDA were not significantly different of the control group?P>0.05?.With the increase of fermented PES feed supplemental ratio,contents of GLU,Ca,K,IgM,TP,ALB,GLB,LDH,HDL,CREA and MDA,as well as activity of AST,ALT,CAT and SOD of the test goats showed an overall increasing trend,contents of UREA showed an decreasing trend,contents of serum P,Na,Cl,TC,TG,LDL,IgA and IgG,as well as activity of CK and GSH-Px were not significantly different between each groups.Above all,the feed quality of PES fermented by the strains screened out in this study can be well improved.The supplemental ratio of 35%of fermented PES feed in diet for Matou goat is the best,and did not affect the animal health.