摘要
对新疆昌吉地区某规模化羊场的疑似绵羊肺炎支原体(MO)感染的病料进行病原分离鉴定,本研究采用羊支原体专用液体培养基和1.0%羊支原体琼脂固体筛选培养基从2份病死羔羊肺脏和13份患病羔羊鼻试子分离得到9株支原体,通过菌落形态学观察,生化试验及特异性PCR方法进行鉴定。结果显示:分离株菌落呈"煎蛋状",Dienes染色菌落中心呈深蓝色,符合支原体菌落染色特征;分离株能发酵葡萄糖,不能水解精氨酸,不分解尿素,明胶液化反应阴性,美兰还原反应阳性符合MO生化特性;参照MO16S r RNA序列进行PCR鉴定,9株培养物均扩增出大小为406 bp的目的片段;生物信息学分析显示,9个分离株与参考株Y98核苷酸同源性均在75%以上,结果表明这9株分离株均为MO,从而确定引起该羊场绵羊呼吸道感染的病原体为MO。本研究结果可为该场绵羊呼吸道病的防控提供诊断依据。
In this study, the suspected Mycoplasma ovipneumoniae（MO） infection samples from the sheep farm of Changji area were conducted to isolate pathogens and then identified by morphological, biochemical and molecular techniques. 9 strains of MO were successfully isolated from 3 lungs and 13 nasal swabs collected from sheep samples suspected of MO infection by Sheep Mycoplasma special liquid medium and 1.0% agar solid sheep Mycoplasma screening medium.There were ＂Omelette shape＂ colonies on the petri dishes,and dienes stain colonies with dark blue center. The isolated could ferment glucose, but could not hydrolyze arginine, nor decomposeurea. It was positive on the Meilan reduction reaction and negative on Gelatin liquefaction reaction consistent with MO. The PCR amplification of the sheep MO showed a specific band of 406 bp by 16 S r RNA. Bioinformatics analysis showed that the nucleotide homology of 16 S r RNA gene of the 9 strains than 75% identity with conference strain Y98 16 S r RNA gene. The results suggested that 9 isolates were MO, which provides diagnostic basis for prevention and control of the sheep respiratory diseases.
出处
《石河子大学学报（自然科学版）》
CAS
2016年第3期291-294,共4页
Journal of Shihezi University(Natural Science)
基金
新疆兵团重大科技项目(2013AA003-3)
