Kilwinski Jochen, Peters Martin, Ashiralieva Ainura, Genersch Elke
Staatliches Veterinäruntersuchungsamt Arnsberg, Zur Taubeneiche 10-12, 59821 Arnsberg, Germany.
Vet Microbiol. 2004 Nov 30;104(1-2):31-42. doi: 10.1016/j.vetmic.2004.08.001.
The bacterial pathogen Paenibacillus larvae subsp. larvae (P. l. larvae), is the etiological agent of American foulbrood, an extremely contagious and disastrous disease of honeybee brood. In case of American foulbrood the destruction of infected colonies is often considered the only workable control measure. Therefore, the ability to diagnose this disease properly is important to prevent unnecessary economic loss to beekeepers. The development of suitable methods for the early and reliable detection of P. l. larvae is hampered by the fact that the two subspecies of Paenibacillus larvae, P. l. larvae and Paenibacillus larvae subsp. pulvifaciens (P. l. pulvifaciens), seem to be indistinguishable by cultural characteristics as well as by PCR protocols. Here we present an extensive analysis of several P. larvae reference strains. We employed conventional culture techniques, morphological and biochemical identification, PCR-based methods and sequencing of the 16S rDNA. We found indeed that P. l. pulvifaciens strain DSM 3615 is indistinguishable from P. l. larvae (DSM 7030). We did not face any problems to discriminate between P. l. larvae and P. l. pulvifaciens strains DSM 8442 and DSM 8443. Therefore, classification of DSM 3615 as type strain of P. l. pulvifaciens seems not to be justified. We propose to reclassify this strain as P. l. larvae. Former problems in differentiating the two subspecies might have arisen from this misclassification. PCR-based methods as well as appropriate biochemical identification systems provide a reliable means for the discrimination between the two subspecies P. l. larvae and P. l. pulvifaciens.
细菌性病原菌幼虫类芽孢杆菌幼虫亚种（P. l. larvae）是美洲幼虫腐臭病的病原体，这是一种极具传染性且对蜜蜂幼虫危害极大的疾病。对于美洲幼虫腐臭病而言，销毁受感染蜂群通常被视为唯一可行的控制措施。因此，正确诊断这种疾病的能力对于防止养蜂人遭受不必要的经济损失至关重要。由于幼虫类芽孢杆菌的两个亚种，即P. l. larvae和粉状幼虫类芽孢杆菌（P. l. pulvifaciens），在培养特征以及PCR检测方法上似乎难以区分，这阻碍了早期且可靠地检测P. l. larvae的合适方法的开发。在此，我们对几种幼虫类芽孢杆菌参考菌株进行了广泛分析。我们采用了传统培养技术、形态学和生化鉴定、基于PCR的方法以及16S rDNA测序。我们确实发现，P. l. pulvifaciens菌株DSM 3615与P. l. larvae（DSM 7030）难以区分。但我们在区分P. l. larvae与P. l. pulvifaciens菌株DSM 8442和DSM 8443时没有遇到任何问题。因此，将DSM 3615归类为P. l. pulvifaciens的模式菌株似乎不合理。我们建议将该菌株重新归类为P. l. larvae。之前区分这两个亚种时出现的问题可能源于这种错误分类。基于PCR的方法以及合适的生化鉴定系统为区分P. l. larvae和P. l. pulvifaciens这两个亚种提供了可靠手段。
