The Therapeutic Effects Research Of Bacillus Subtilis,Lactobacillus Fermentum Strain And Pediococcus Acidilactici Compound Probiotics On Rabbit Endometritis

[Objective]Endometritis is a common disease in postpartum,and most of them are recessive,brought a certain threat to the development of livestock breeding.Over the years,researchers have been seeking effective and safe treatment methods,to explore the therapeutic effect of endometritis.Probiotics are compounded,is still non-toxic and safe,it will not cause drug residues,enhanced the immunity of animal body.This study is to explore treatment effect of compound probiotics on endometritis.[Materials and Methods]Experiment 1:Taken the vaginal secretions of dairy cows,taken anaerobic and aerobic way of cultivation,respectively,after separation and purification,staining microscopy,physiological and biochemical reaction,analysis of 16SrDNA sequences and homology comparison to determine strains.Experiment two:(1)The optimal growth conditions of each strain were determined by comparing the growth curve,the culture pH,the inoculation amount of the strain and the volume of the liquid culture;(2)The effect of antibacterial was determined by Oxford cup method in vitro,three strains can or cannot grow in the same environment(3)A total of 40 Kunming SPF mice were randomly divided into 4 groups.The control group(K),Bacillus subtilis group(S1),Lactobacillus fermentzum strain group(S2)and Pediococcus acidilactici group(S3),7d after,organ coefficient were calculated to evaluate strains of safety.Experiment three:(1)20 Kunming SPF mice were randomly divided into 2 groups,experimental group(probiotics)and control group,7d after,the number of the main inflammatory cells in blood was detected and the organ coefficient was calculated to evaluate the safety of compound probiotics.(2)The 15 female rabbits were divided into three groups:control group,infection group and treatment group(n = 3).The rabbit endometritis model was established and treated by intrauterine infusion of pathogenic bacteria and compound probiotics.Times of administration were three times,once every other day.Observed clinical symptoms,detected urine,the classification count of white blood cell,uterine morphology and pathological sections of uterus,detected antibody levels of serum and level of inflammatory related factors to evaluate the therapeutic effect of the endometritis.[Results]1?After the separation,purification,microscopic examination,physiological and biochemical reaction,analysis of 16SrDNA sequence and homology,Two strains of probiotics were isolated in study,Bacillus subtilis and Lactobacillus fermentum strain.2?The optimal culture conditions of the three strains were determined by measuring the biological characteristics of the strains.All strains were cultured at 37 ?.Bacillus subtilis with inoculum amount of 6%20 ml pH =6.8 LB liquid culture medium,18 h,its activity was the strongest;Pediococcus acidilactici in inoculum amount of 2%30 mL pH =6.4 MRS liquid culture medium,24 h anaerobic fermentation;Lactobacillus fermentum strain in inoculation amount of 5%30 mL pH=6.4 MRS liquid culture medium,24 h anaerobic fermentation.In vitro antibacterial activity of three strains was detected by Oxford cup method.The results showed that:Three strains had obvious antibacterial effect.By detecting the compatibility among three strains,they can grow in the same environment,no antagonism.Through the toxicity test,it was found that the three strains does not cause the pathological changes of the main organs of mice,and there was no significant difference in organ coefficient(p>0.05).The results showed that three strains were safe when used alone.3?Through the toxicity test,it was found that the combination of three strains does not cause the pathological changes of the main organs of the mice,and there was no significant difference in organ coefficient(p>0.05).Blood routine examination showed that there was no significant difference in the number of inflammatory cells(p>0.05).The results showed that the three strains were safe when were compounded.4?After treatment with compound probiotics,the animals had good mental state,and urine examination results were negative with silver nitrate method.Compared with the infection group,the number of inflammatory cells of the treatment group were significantly reduced,and the difference was significant(p<0.01,p<0.05).The uterine morphology was observed,and the degree of swelling and congestion was relieved.Observed the pathological sections of uterine tissue,compared with infection group,endometrial epithelial cells of uterine of treatment group regularly arranged,infiltration of neutrophils significant decreased,uniform distribution of uterine gland,vacuolization degree was significantly reduced.5?Tested antibody level and main inflammatory factor levels by using the method of ELISA,compared with infection group,the levels of IgA,IgG and IgM of the treatment group in serum were significant higher,both showed significant difference(p<0.01,p<0.05);the levels of inflammatory cytokine IL-1? and CRP of treatment group were significant decreased,showed significant difference(0.01<p<0.05).[Conclusion]Bacillus subtilis,Lactobacillus fermentum strain and Pediococcus acidilactici after compound is safe can improve immunity of animals,body and relieve inflammation.It has therapeutic effect on endometritis.