The prevalence of the honeybee brood pathogens Ascosphaera apis, Paenibacillus larvae and Melissococcus plutonius in Spanish apiaries determined with a new multiplex PCR assay

Authors


  • Funding Information This study was supported by funding from Junta de Comunidades de Castilla-La Mancha (Consejería de Agricultura and Consejería de Educación), INIA (RTA2005-152 and RTA 2008-00020-C02-01, FEDER funding) and MARM-FEAGA (Programa Nacional Apícola 2011–2013). INCRECYT, European Social Funds.

For correspondence. E-mail mhiges@jccm.es; Tel. (+34) 949 250026; Fax (+34) 949 250176.

Summary

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.

Ancillary