Isolation And Identification Of Mycoplasma Bovis And Epidemiological Surver In Ning-xia

In recent years,Mycoplasma bovis was showing trends in our country,it also showed an increasing trend in Ning-xia,in this study,Ning-xia Mycoplasma bovis were investigated separation,identification and epidemiology,and made improvements susceptibility test and media on it.mainly in the following conclusions.(1)On november 2013,introduced some cows in a dairy farm in Ning-xia from other dairy cows and calves,a few weeks the similar adult cow disease continue to occur,for the diagnosis of pathogens,disease sterile samples of material collected for bacterial and mycoplasma culture and separated by Gram stain,colony morphology,the results show:Bacteria isolated from E.coli,Streptococcus,bacteria and other bus,isolated three strains of mycoplasma were named as NP 1,NP 2,NP 3,to three mycoplasma bovis isolates using specific PCR identification and 16S rRNA sequence alignment,NJ method using three isolates of 16S rRNA sequences of representative strains of mycoplasma establish phylogenetic tree,the results confirmed that the epidemic is causing the field the main pathogen mycoplasma bovis,and mixed infections with other bacteria.(2)Mycoplasma bovis exhibit epidemic in North America,Europe and other countries and regions,since 2008,Chinese first reported in cattle caused by it So far,a number of provinces have reported in the region,in order to master the dairy farm in Ning-xia and infection in cattle farm,ELISA test kit and specific PCR were used to detect the Ning-xia region between 1150 sera from November 2013 to December 2014,acquisition of mycoplasma bovis antibody levels and 88 tissue samples specific gene.The results showed that the detection of bovine serum in 1380,detected 319 positive,the positive rate was 23.12%,percent of individuals in the detection of 89 diseased samples were detected,positive number 53 overall,positive rate was 59.55%.(3)Because Mycoplasma bovis nutrients needs its own synthesis was weak,higher in vitro culture conditions and its nutrient needs,and the medium is expensive,in order to improve the efficiency of Mycoplasma bovis culture,we explored the Mycoplasma bovis in liquid the optimum pH value of the growth medium,the optimal serum concentrations and optimum concentration of yeast extract,in order to further improve the efficiency of the original body of the culture medium and modified Thiaucourt's ability to inhibit bacteria growth,in the medium deoxygenated sodium ribose nucleotides,beef heart soup and bovine lung digestion,inhibit the growth of gram-positive bacteria crystal violet,using the viable count method,the bacterial protein assay,OD values were measured,the results confirm the improvement Thiaucourt's the above mentioned substances can inhibit the growth of the added efficiency mycoplasma bovis,while improving the ability of the medium to inhibit bacteria.After the modified Thiaucourt's liquid medium was inoculated Mycoplasma bovis discoloration fastest time from 2d becomes 1d,improved Thiaucourt's solid medium after inoculation Mycoplasma bovis can be observed under the microscope colonies fastest time from 3d becomes 2d.(4)Mycoplasma bovis in domestic prevalence and incidence of the more serious,and the overuse of antibiotics leads to resistance enhanced it,commonly DST is not available for the detection of it susceptibility sensitivity.To this end,we selected three alkaloids(oxymatrine,matrine,matrine),Mycoplasma bovis was measured in these three alkaloids minimum inhibitory concentration(MIC),while using micro-dilution method,we identified susceptibility kit made improvements apply to both the initial identification and susceptibility testing of it,At the same applies to preliminary identification and susceptibility testing of it,it achieved the identification and susceptibility test simultaneously,enhance the efficiency of diagnosis and treatment of disease.