镰形扇头蜱唾液腺、中肠和卵的菌群结构

Identification of the bacteria in the salivary glands,midgut and egg from Rhipicephalus haemaphysaloides

采用PCR-DGGE技术分析了镰形扇头蜱雌成蜱中肠内容物、唾液腺、虫卵的菌群结构。无菌条件下采集镰形扇头蜱雌成蜱中肠内容物、唾液腺、虫卵,提取细菌总DNA;以通用引物扩增细菌16SrDNA V3区;DGGE电泳,回收、克隆、测定DGGE优势条带。结果表明,饱血和半饱血雌蜱中肠菌群结构相同,中肠内容物、唾液腺和虫卵部分细菌相同;9条明显条带的序列分别与柯克斯体属(Coxiella sp.)、假单胞菌属(Pseudomonas sp.)芽孢杆菌属(Bacillus sp.)、葡萄球菌属(Staphylococcus sp.)、涅斯捷连科菌属(Nesterenkonia)、共生菌(Symbiotic bacteria)和未培养土壤细菌的16SrDNA V3区序列高度相似。分离到2株细菌,为模仿葡萄球菌和蜡样芽孢杆菌。

The bacteria in the salivary glands,midguts and eggs from Rhipicephalus haemaphysaloides were analyzed with PCR and denaturing gradient gel electrophoresis（PCR-DGGE）.The analysis process were as follows：collecting the salivary glands,midguts and eggs in sterile conditions,extracting the bacterial total genomic DNA,amplifying the 16 SrDNA V3 with universal primers,DGGE electrophoresis,recycling,cloning and sequencing the selected bands.The results showed that the midgut microbiota of the engorged ticks and the semi-engorged one were in the same,and the florae of the female R.haemaphysaloides salivary glands,midguts and eggs were different.Nine distinct DGGE bands were found using PCR-DGGE method,and they belonged to Coxiellasp.,Pseudomonas sp.,Bacillus sp.,Staphylococcus sp.,Nesterenkonia,Symbiotic bacteria and Uncultured soil bacterium,respectively.Two distinct strains were isolated using culturedependent method,and they were Staphylococcus simulans and Bacillus cereus.