Screening Of Clostridium Butyricum And Its Application In Growing Pigs

Microbial anaerobic culture was used in this experiment,a strain of Clostridium butyricum was isolated and screened from pig manure;The strain was preliminarily identified by the morphological characteristics of the colony and the microscopic observation of gram staining;Further experiments were carried out,including sugar fermentation experiment,gelatin liquefaction,starch hydrolysis,litmus milk and other physiological and biochemical experiments,16SrDNA sequence analysis and identifica-tion.Then the growth characteristics and antibacterial activity in vitro were studied.The pigs were grown as guinea pigs,Added in the diet with the research on the basis of growing pigs Clostridium butyricum acid and butyric acid Bacteria,Yeast,Bacillus and composite in enterococcus bacteria fermentation biological material for its growth performance,diarrhea rate,serum biochemical index,the influence of oxidation resistance and immunity performance,In addition,the quantity of E.coli、Lactobacillus and Salmonella were determined in each group.Screening and identification of Clostridium butyricum:a strain of Clostridium butyricum was screened from pig manure and named Cbutyricum C3.Determination of the growth characteristics of Clostridium butyricum:The optimal growth temperature of C.butyricum C3 is 34℃,The optimum initial pH is 7.0,The bacteria suspension in 90 ℃ water bath 1 min survival rate was 31.7%.Clostridium butyricum was cultured for 4 h in RCM and began to proliferate rapidly,reaching thepeak at 8 h;The pH value of the RCM liquid medium gradually decreased with the incubation time.After 12 h,the pH gradually stabilized around 4.6.C.butyricum C3 was resistant to the glycopeptide antibiotics such as amikacin and gentamicin,and was less sensitive to penicillin,ciprofloxacin,norfloxacin and compound sulfamethoxazole.In vitro antibacterial properties:The fermentation supernatant of Clostridium butyricum C3 has inhibitory effects on Staphylococcus aureus,White staphylococcus,Salmonella typhimurium and E.coli of piglets,The inhibition effect of enterococcus and pseudomonas was not obvious.The bacteriostatic effect of crude extract of C.butyricum C3 is better than that of supernatant.Study on the application of Clostridium butyricum in growing pig diet:A total of 160 pigs with a healthy weight of 44±2.4kg were selected,Single factor randomized trial design was adopted,They were divided into 4 groups:Control group C,Test TPF,Test FPF and Test CB,Two repetitions per group,20 pigs per repeat.Control group C feeding growing pig feed;Test TPF,Test FPF and Test CB were added Biological feed A,Biological feed B and Clostridium butyricum powder on the basis of feeding compound feed of growing pigs.The pH value of biological feed A was 4.2 and the water content was 44%.The contents of Yeasts,Bacillus and Enterococcus faecalis were 16×108 cfu/g,13×108 cfu/g and 32×108 cfu/g respectively.The pH value of biological feed B was 4.3 and the water content was 42%.The contents of Yeasts,Bacillus,Enterococcus faecalis and Clostridium butyricum were 2×108 cfu/g、10×108 cfu/g、33×108 cfu/g and 20×104 cfu/g respectively.The results show that:Growth performance:In the test of 1-34 day,average daily gain,feed intake and feed conversion ratio were not significantly different(P>0.05);On the 17～34 day of experiment,average daily gain and average daily intake increased.Microbial flora in feces:Compared with the Control group C,the number of Escherichia coli in the Test TPF,Test FPF and Test CB decreased by 6.5%(P>0.05)、21.8%(P<0.05)and 17%(P<0.05);Compared with TFP,the number of E.coli in FPF was significantly reduced by 16.4%(P<0.05);The number of lactic acid bacteria increased by 13.2%、10.7%and 13.0%respectively,and the difference was significant(P<0.05).Serum enzyme:Compared with the Control group C,the serum alanine transaminase in the Test TPF,Test FPF and Test CB decreased by 4.7%(P>0.05)、16.3%(P<0.05)and 25%(P<0.05),respectively;In comparison with TPF,the Test FPF and the Test CB serum were decreased by 12.2%(P<0.05)and 21.3%(P<0.05),respectively.Immune performance:Compared with the Control group C,the Test TPF,Test FPF and Test CB immunoglobulin A(IgA)increased by 29.6%,35.2%and 16.9%respectively,with no significant difference(P>0.05);Immunoglobulin G(IgG)increased by 0.32%(P>0.05)、9.6%(P>0.05)and 20.4%(P<0.05),respectively.Antioxidant properties:Compared with the Control group C,the Test TPF,Test FPF and Test CB glutathione peroxidase increased by 2.4%(P>0.05)、3.4%(P<0.05)and 9.5%(P<0.05),the total superoxide dismutase T-SOD increased by 0.3%、8.2%and 9.2%,respectively,the difference was significant(P<0.05);Compared with the Test TPF,the serum GSH-PX in the Test FPF and the Test CB increased by 2.5%(P>0.05)and 8.5%(P<0.05),and the serum T-SOD increased by 7.9%and 8.9%,respectively,and the difference was significant(P<0.05).In summary:In this experiment,we screened a Clostridium butyricum from pig feces.And determine the optimum temperature,the optimal initial pH and the sensitivity to antibiotics.C.butyricum C3 has inhibitory effect on Staphylococcus aureus,White Salmonella,Salmonella typhimurium and Escherichia coli of piglets.Clostridium butyricum and mixed fermentation organisms containing Clostridium butyricum were added to the basal diet of growing pigs,It can significantly decreased the number of Escherichia coli in feces and increase the number of lactic acid bacteria in feces.At the same time,it can significantly increase the content of IgG in serum and enhance the activity level of GSH-PX and T-SOD.