title: The prevalence of the honeybee brood pathogens Ascosphaera apis, Paenibacillus larvae and Melissococcus plutonius in Spanish apiaries determined with a new multiplex PCR assay.
abstract: 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.
authors.affiliation: Bee Pathology Laboratory, Centro Apícola Regional (CAR), Junta de Comunidades de Castilla La Mancha, 19180, Marchamalo, Spain.
authors.forename: Encarna
authors.initials: E
authors.lastname: Garrido-Bailón
authors.forename: Mariano
authors.initials: M
authors.lastname: Higes
authors.forename: Amparo
authors.initials: A
authors.lastname: Martínez-Salvador
authors.forename: Karina
authors.initials: K
authors.lastname: Antúnez
authors.forename: Cristina
authors.initials: C
authors.lastname: Botías
authors.forename: Aránzazu
authors.initials: A
authors.lastname: Meana
authors.forename: Lourdes
authors.initials: L
authors.lastname: Prieto
authors.forename: Raquel
authors.initials: R
authors.lastname: Martín-Hernández
mesh_terms: D000818:Animals; D001203:Ascomycota / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001516:Bees / Q000382:microbiology*; D056569:Enterococcaceae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D056507:Paenibacillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D013030:Spain
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