[Reclassification of the four China isolated strains of the pathogen for contagious caprine pleuropneumonia].

Contagious caprine pleuropneumonia (CCPP) is caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp). The aims of this study were to identify 4 Chinese isolated strains employing molecular methods and to determine the appropriate subspecies classification of these strains. Three genome fragments (A, B and C) from each strain were amplified and then transformed into plasmids. The inserted fragments were sequenced and analyzed by comparison with six members of the Mycoplasma mycoides cluster. Cleavage of the PCR products of the 4 strains with PstI yielded three fragments 548, 420 and 128bp in length, just like strain F38. The other M. mycoides cluster members had only 2 fragments of 428 and 128bp. Homology analysis of fragment B indicated that the 4 strains exhibited 99.5% homology with Mccp reference strain F38, 98.9% with M. capricolum subsp. Capricolum (Mcc) strain California Kid, and only 95.4% with Mmc strain ZZ. In fragment C, the 4 strains had 67.4% - 67.6% homology with Mmc PG3, 95.1% -98.6% with Mcc strains 8601-50 and California Kid, 99.6% - 99.8% with Mccp strains 97097ET, Gabes and F38. The analysis revealed that 4 pathogeny strains, 87001, 87002, 367, 1653, isolated from China are more closely related to Mccp than to Mcc. Therefore the pathogeny of CCPP in China should be reclassified as Mccp.

Antimicrobial susceptibility and molecular characterization of macrolide resistance of Mycoplasma bovis isolates from multiple provinces in China.

Mycoplasma bovis has spread widely throughout the world via animal movement and has become an important pathogen of bovine respiratory disease. However, the minimum inhibitory concentrations of antimicrobials for Mycoplasma bovis have not been studied in China. The objective of this study was to determine the prevalence and antibiotic resistance of Mycoplasma bovis isolated from young cattle with respiratory infection in China. Mycoplasma bovis was detected in 32/45 bovine respiratory infection outbreaks at beef farms in 8 provinces in China. The isolates were susceptible or had medium sensitivity to ciprofloxacin, enrofloxacin and doxycycline, but were frequently resistant to macrolides (13/32, 41%). An A2058G (Escherichia coli Numbering) mutation located in the rrnA operon in domain V of 23S rRNA was observed in strains that were resistant to macrolides. This single mutations at the rrnA operon in domain V of 23S rRNA may play an important role in the resistance of Mycoplasma bovis strains to macrolides.

[Expression, purification and antigen activity analysis of the N-terminal domain of lipoprotein LppQ of Mycoplasma mycoide subsp. mycoides SC].

The gene sequence coding the N-terminal domain of LppQ was amplified from Mycoplasma mycoides subsp. mycoides SC (MmmSC) HVRI X strain by PCR using special primers and was cloned into the EcoR I /Sal I sites of pET32a vector to construct the expression recombinant plasmids. The recombinant plasmids were indentified by restriction digestion, PCR and sequence analysis. The gene was overexpressed in Escherichia coli BL21 (DE3) host cell and the soluble protein was purified with Ni-NTA His. Bind purification kits. The amount of recombinant protein reached 53.7% of the total mass of bacterial protein. The purity of recombinant protein reached to over 95 %. The antigen activity of the purified protein was examined with Western blot analysis. The purified protein reacted strongly with the standard positive serum and didn't react with the negative sera of contagious bovine pleuropneumonia(CBPP).

Isolation and genetic characterization of bovine parainfluenza virus type 3 from cattle in China.

Bovine parainfluenza virus type 3 (BPIV3) is one of the most important of the known viral respiratory pathogens of both young and adult cattle. However BPIV3 has not been detected or isolated in China prior to this study. In 2008, four BPIV3 strains were isolated with MDBK cells from cattle in China and characterized by RT-PCR, nucleotide sequence analysis, transmission electron microscope observation, hemadsorption and hemagglutination tests. Nucleotide phylogenetic analysis of partial hemagglutinin-neuraminidase (HN) gene for four isolates and the complete genome for the SD0835 isolate implicated that the four Chinese BPIV3 strains were distinct from the previously reported genotype A (BPIV3a) and genotype B (BPIV3b) and might be a potentially new genotype, which was tentatively classified as genotype C (BPIV3c). This is the first study to report the isolation and genetic characterization of BPIV3 from cattle in China.