Detection of tau-fluvalinate resistance in the mite Varroa destructor based on the comparison of vial test and PCR-RFLP of kdr mutation in sodium channel gene
Language English Country Netherlands Media print-electronic
Document type Comparative Study, Journal Article
Grant support
QJ1530148
Národní Agentura pro Zemědělsk Vzkum
PubMed
30810851
DOI
10.1007/s10493-019-00353-9
PII: 10.1007/s10493-019-00353-9
Knihovny.cz E-resources
- Keywords
- Acaricide, Apiary, Honey bee, Restriction, Varroa destructor,
- MeSH
- Acaricides pharmacology MeSH
- Biological Assay methods MeSH
- Densitometry methods MeSH
- Drug Resistance genetics MeSH
- Nitriles pharmacology MeSH
- Polymerase Chain Reaction methods MeSH
- Polymorphism, Restriction Fragment Length MeSH
- Pyrethrins pharmacology MeSH
- Varroidae drug effects genetics MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
- Names of Substances
- Acaricides MeSH
- fluvalinate MeSH Browser
- Nitriles MeSH
- Pyrethrins MeSH
Varroa destructor is the major cause of honey bee (Apis mellifera) colony losses. Mite control is limited to several miticides. The overuse of tau-fluvalinate has resulted in resistance via a knockdown resistance (kdr) mutation in the sodium channel gene NaVChs (L925V/I/M). In this study, we used the discriminating concentration of tau-fluvalinate (0.25 µg/mL) to detect the resistance of mites in a bioassay. Further, we verified the presence of the kdr mutation in mites from the bioassay via PCR amplification of a fragment of the voltage-gated sodium channel gene (NaVCh), restriction fragment length polymorphisms (RFLPs), and densitometry analyses in pools of surviving or dead mites. Resistance values corresponding to the densitometry of the resistant allele were related to mite survival. In the vial test, the survival of the control group was significantly higher (70.4%) than that of the tau-fluvalinate-treated group (34.3%). Mite survival in the vial test was significantly correlated with the mean proportion of resistance values. Individuals that died after tau-fluvalinate application exhibited an average resistance value of 0.0783, whereas individuals that survived exhibited an average resistance of 0.400. The concentration of tau-fluvalinate in the vials was checked using high performance liquid chromatography under different temperatures and exposure times, and indicates that the stability of tau-fluvalinate stored in the refrigerator (4 ± 1 °C) is at least 14 days. PCR-RFLP of the NaVCh gene fragment verified that the vial test is a suitable, rapid, and cost-effective method for the identification of tau-fluvalinate resistance based on kdr mutation in V. destructor in apiaries.
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