Hui Lijing, Zhong Zhiping, Hu Bing, Yang Bing, Ji Yanling, Yu Hanshou
Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.
Wei Sheng Wu Xue Bao. 2010 Oct;50(10):1366-72.
To investigate the kinds and characteristics of spiroplasma in honeybees, as well as to study the taxonomy and transmission of honeybee spiroplasma under natural conditions.
We examined the morphology of spiroplasma isolates by dark field and transmission electron microscopy and studied the biological characteristics by using conventional culture-dependent methods and molecular biology and serological methods.
Three spiroplasma isolates were obtained from healthy Apis mellifera. All isolates exhibited helicity during their growth phase, with one isolate (MF0905) being shorter and having less helicity. This isolate also differed from the other two (MF0903 and MF0904) in having larger colonies with an irregular margin instead of being round. In addition, isolate MF0905 could not hydrolyze arginine whereas MF0903 and MF0904 could. All three isolates could use glucose and D-fructose as a carbon source but did not hydrolyse urea. Phylogenetic analysis based on the 16S rDNA, ITS and the rpoB gene showed that MF0903 and MF0904 had a close relationship with Spiroplasma melliferum, and MF0905 was close to S. clarkii. Serological studies including the growth inhibition test, metabolic inhibition test and deformation test gave the same result as the phylogenetic analysis.
The spiroplasma isolate MF0905 might be S. clarkii and other two isolates were S. melliferum. This result indicated that Spiroplasma melliferum is not the only spiroplasma species in honeybees in China.
调查蜜蜂体内螺旋体的种类和特征，以及研究自然条件下蜜蜂螺旋体的分类和传播。
我们通过暗视野显微镜和透射电子显微镜检查螺旋体分离株的形态，并使用传统的依赖培养的方法、分子生物学方法和血清学方法研究其生物学特性。
从健康的意大利蜜蜂中获得了三株螺旋体分离株。所有分离株在生长阶段均呈现螺旋状，其中一株（MF0905）较短且螺旋度较小。该分离株与其他两株（MF0903和MF0904）的不同之处还在于其菌落较大且边缘不规则，而非圆形。此外，分离株MF0905不能水解精氨酸，而MF0903和MF0904可以。所有三株分离株都可以利用葡萄糖和D-果糖作为碳源，但不能水解尿素。基于16S rDNA、ITS和rpoB基因的系统发育分析表明，MF0903和MF0904与蜜蜂螺旋体关系密切，而MF0905与克拉克螺旋体关系密切。包括生长抑制试验、代谢抑制试验和变形试验在内的血清学研究结果与系统发育分析一致。
螺旋体分离株MF0905可能是克拉克螺旋体，其他两株分离株是蜜蜂螺旋体。这一结果表明，蜜蜂螺旋体不是中国蜜蜂体内唯一的螺旋体物种。
