Bee Pathology Laboratory, Centro Apícola Regional (CAR), Junta de Comunidades de Castilla La Mancha, 19180, Marchamalo, Spain.
Microb Biotechnol. 2013 Nov;6(6):731-9. doi: 10.1111/1751-7915.12070. Epub 2013 Aug 6.
The microorganisms Ascosphaera apis, Paenibacillus larvae and Melissococcus plutonius are the three most important pathogens that affect honeybee brood. The aim of the present study was to evaluate the prevalence of these pathogens in honeybee colonies and to elucidate their role in the honeybee colony losses in Spain. In order to get it, a multiplex polymerase chain reaction (PCR) assay was developed to simultaneously amplify the16S ribosomal ribonucleic acid (rRNA) gene of P. larvae and M. plutonius, and the 5.8S rRNA gene of A. apis. The multiplex PCR assay provides a quick and specific tool that successfully detected the three infectious pathogens (P. larvae, M. plutonius and A. apis) in brood and adult honeybee samples without the need for microbiological culture. This technique was then used to evaluate the prevalence of these pathogens in Spanish honeybee colonies in 2006 and 2007, revealing our results a low prevalence of these pathogens in most of the geographic areas studied.
影响蜜蜂幼虫的三种最重要的病原体是节肢动物芽孢杆菌、蜂房芽孢杆菌和花翅实蝇球孢菌。本研究旨在评估这些病原体在西班牙蜜蜂群体中的流行程度，并阐明它们在蜜蜂群体损失中的作用。为此，开发了一种多重聚合酶链反应（PCR）检测方法，以同时扩增蜂房芽孢杆菌和花翅实蝇球孢菌的 16S 核糖体核糖核酸（rRNA）基因，以及节肢动物芽孢杆菌的 5.8S rRNA 基因。该多重 PCR 检测方法提供了一种快速而特异的工具，可成功检测幼虫和成年蜜蜂样本中的三种传染性病原体（蜂房芽孢杆菌、花翅实蝇球孢菌和节肢动物芽孢杆菌），而无需进行微生物培养。然后，我们使用该技术评估了 2006 年和 2007 年西班牙蜜蜂群体中这些病原体的流行程度，结果表明，在大多数研究的地理区域，这些病原体的流行率都较低。
