A new detection method for a newly revealed mechanism of pyrethroid resistance development in Varroa destructor.

Aneta Strachecka,Grzegorz Borsuk,Krzysztof Olszewski,Jerzy Paleolog

doi:10.1007/s00436-015-4627-4

Abstract

The Varroa destructor mite has recently displayed an ever increasing resistance to new drugs, contributing to CCD proliferation. This work was aimed at determining new viable methods for identifying the pyrethroid resistance of V. destructor and DNA methylation in resistant and sensitive mites. DNA was extracted from Varroa mites. Nucleotide changes in the DNA of pyrethroid-resistant, pyrethroid-sensitive, and control mites were identified with polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) in the case of five mitochondrial gene fragments. More bands were observed in the drug-resistant mites than in the other two groups. Sequencing confirmed these observations. Decreased global DNA methylation levels were observed in the pyrethroid-resistant mites. There exists a previously undescribed mechanism of pyrethroid resistance development in Varroa mites. The PCR-SSCP methods can be considered and further developed as useful tools for detecting V. destructor resistance.

Highlights

Varroa destructor parasite (Fig. 1) is the crucial, worldwide problem of apiculture
The aim of this study was (1) to develop a new method for detecting pyrethroid resistance of V. destructor based on polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) involving five mitochondrial genes
polymerase chain reaction single-strand conformation polymorphism (PCRSSCP) is a simple and effective technique detecting changes in the nucleotide sequence of PCR products (2) to reveal the existence of a yet undescribed mechanism of V. destructor pyrethroid resistance connected with nuclear DNA methylation

Summary

Introduction

Varroa destructor parasite (Fig. 1) is the crucial, worldwide problem of apiculture. It is believed to be one of the mainThe resistance within the parasite populations has a multifactorial character. Yukioka et al (1998) and Van Leeuwen et al (2008) discovered that Varroa resistance to this acaricide is associated with mutations of mitochondrial genes. Wang et al (2002) and Liu et al (2006) described the third resistance mechanism: Taufluvalinate causes point mutations in mitochondrial DNA (mtDNA) and blocks sodium channels, which increases Varroa tolerance of this pyrethroid. Such mutations result in an exchange of Parasitol Res (2015) 114:3999–4004

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