Isolation Identification Of Mycoplasma Ovipneumoniae And Genomic Sequence Determination And Analysis

Mycoplasma ovipneumoniae (MO)is considered an emerging veterinary pathogen causing intersticial and proliferative pleuropneumonia in sheep and goats。The disease appeared to be spreading in the world, which caused great sufferings and losses to the sheep husbandry。So people increasingly pay attention to the study on the disease infected by Mycoplasma ovipneumoniae。 A deep research was made on MO with isolation and identification of pathogen, method of diagnosis and prevention and trentment to provide the scientific basis for controlling it in production。1、12Mycoplasma strains were isolated from33pneumonia lungs and pleural effusion samples of probably infectious pneumonia of sheep and goats come from the siziwangqi and dalateqi areas in Inner Mongolia. Lung tissue samples after cut and grinded under sterile condition were inoculated on the modified Hayflick medium;pleural effusion after filtered by micr-opore filter of0.22μm aperture were inoculated on the modified Hayflick medium. The results of staining microscopic examination, biochemistry experiments and serologic tests suggested that the isolates and MO pattern strain Y98were the same serotype.2、The16SrRNA genes with a fragment of700bp DNA in length were amplified from the separated strain D13D27and XJ and the type strain Y98by PCR using the primer pair designed according to the reported16SrRNA gene sequences of MO.They were cloned and sequenced subsequently。 Comparative analysis of these16SrRNA gene sequences showed that D13, D27and XJ strain shared97.7%,98.6%and100%identity in nucleotide sequence with type strain MO respectively。 The results showed that the isolates belonged to MO。 And MO was the primary pathogen of sheep and goats mycoplasmal pneumonia。3、In this paper, a method is proposed for generating MO cell growth curve by determining the absorption value at450nm of liquid culture with Enzyme-labelled meter。 Cell growth curves of the isolate strain D13,27, XJ and the type strain Y-98were generated in the modified Hayflick medium, the modified KM2medium, the modified FM4medium, the modified Hartley smedium and Mycoplasma Broth Base added Mycoplasma Supplement G respectively。The results showed the most suitable medium for MO was the modified Hayflick medium. 4、The effects of12antibacterial drugs including telithromycin,florfenicol, kanamycin, cillimycin, enrofloxacin, neomycin, tetracycline, streptomycin, erythromycin, gentamicin, and roxithromycin on the isolates in vitro were showed by using the method of micro checkerboard titration。 The results showed that kanamycin, telithromycin, neomycin, enrofloxacin, florfenicol and cillimycin were highly inhibitory on the isolates。 And the6antibacterial drugs were the first choice for the infection of MO in the siziwangqi and dalateqi areas in Inner Mongolia.5、 We sequenced the complete genome of MO strain XJ。 The analyses of gene families, genome annotation and gene function prediction were performed。 This provides a scientific basis for disease prevention and prophylactic vaccine development in the future。...