Detection of acute bee paralysis virus by RT-PCR in honey bee and Varroa destructor field samples: rapid screening of representative Hungarian apiaries

Abstract

-A two years survey was undertaken to determine the occurrence of Acute Bee Paralysis Virus (ABPV) in field samples of adult bees and the parasitic mite Varroa destructor. To detect the viral nucleic acid we used polymerase chain reaction following reverse transcription (RT-PCR). We demonstrated the presence of ABPV RNA in 14 from 114 seemingly healthy colony samples collected from Hungarian honey bees. The investigation revealed that two third of the apiaries were infected with ABPV at a 12.2% infection rate. In seven other apiaries out of eight investigated (87.5%) the presence of the virus was detected from colonies following a sudden collapse; these colonies were simultaneously infected with Nosema apis or infested with Varroa destructor. Virus specific nucleic acid was also identified in the mites collected from two apiaries falling into the latter category. The amplicon of RT-PCR was sequenced and the nucleic acid sequence was aligned to the complete ABPV sequence deposited in the GeneBank database revealing a 93% homology. acute bee paralysis virus / polymerase chain reaction / Varroa destructor / Nosema apis / honey bee

A real-time PCR based survey on acute bee paralysis virus in German bee colonies

“…Previous studies diagnosed significant ABPV infections post mortem or after the manifestation of severe symptoms (Ball, 1985;Ball and Allen, 1988;Bakonyi et al, 2002;Berenyi et al, 2006). Our prospective approach confirms the data about the relevance of ABPV from retrospective surveys.…”

“…While several protocols for a qualitative detection of ABPV exist (Benjeddou et al, 2001;Bakonyi et al, 2002;Tentcheva et al, 2004;Berenyi et al, 2006;Grabensteiner et al, 2007) only few approaches for the quantification of ABPV have been developed. Allen et al (1986) developed an indirect ELISA for ABPV, but the main drawback of serological quantification for bee virus research is the limited availability of antisera and the inconsistency between batches of antisera preparations.…”

“…No data are available about the sample size that is required to detect ABPV with a given probability. In previous studies, a sampling size of up to 60 worker bees was used (Bailey et al, 1981;Ball, 1985;Nordström et al, 1999;Bakonyi et al, 2002;Cox-Foster et al, 2007). Tentcheva et al (2004) and Gauthier et al (2007) used pools of 100 individuals.…”

“…Extracts of heads from field samples were analysed by end-point PCR according to a published protocol (Bakonyi et al, 2002) with slight modifications. A mixture of reverse transcriptases and taq polymerases was used (Qiagen One Step RT-PCR kit).…”

Integration of Molecular and Genetic Methods in the Investigation of the Biodiversity of the Honey Bee (Apis Mellifera L., Hymenoptera: Apidae) and Diagnosis of Diseases of Economic Importance Affecting the Bees and their Brood in Bulgaria

“…Genetic variability detection was performed also based on microsatellite marker profiles [15,18]. Varroa species are the main vectors for horizontal transfer of viruses in bee colonies [19][20][21] and a possible vectors for horizontal transmission of some bacteria (Wolbachia) [22]. Because of that most of the researches are focused on molecular identification of viral infection and their association with mites as vectors.…”