Proteomic analysis of Bayvarol^® resistance mechanisms in the honey bee parasite Varroa destructor

Abstract

The haemophagous mite, Varroa destuctor is one of the most dangerous threats to the Western honey bee, Apis mellifera. Varroa mites parasitize the larval and adult stages of the honey bee and can have devastating effects on the health of the individual bee and colony. In recent years, varroa have shown resistance to the pyrethroid group of insecticides, including Bayvarol ® which has flumethrin as the active ingredient. In the work presented here, changes in the expressed proteomes of mites, either sensitive or resistant to Bayvarol ® were observed using 2D-SDS-PAGE and shotgun label-free proteomics. A number of detoxification proteins (e.g., glutathione-s-transferase, flavin-containing monooxygenase) were present at higher levels in the resistant mites, as were some proton pumping proteins (e.g., Na+/K+ ATPase alpha and beta subunit, E1-E2 ATPase protein). A decrease in the abundance of 12 cuticle proteins in the resistant mites was observed indicating that alteration to cuticle structure could be a potential resistance mechanism. A number of structural proteins such as myosin and alpha tubulin were expressed at higher levels in the resistant mites, which could indicate a change to the intracellular structure of the cuticle barrier or a change in the cell shape/surface, rather than the addition of extra cuticle proteins. The results presented here indicate higher levels of protein associated with cellular detoxification in Bayvarol ® -resistant varroa mites.Análisis proteó mico de los mecanismos de resistencia a Bayvarol ® en el parásito de la abeja de la miel Varroa destructor El ácaro hemó fago, Varroa destuctor es una de las amenazas más peligrosas para la abeja occidental de la miel, Apis mellifera. Los ácaros varroa parasitan los estadios larvarios y adultos de la abeja de la miel y pueden tener efectos devastadores sobre la salud de la abeja y la colonia individual. En los últimos años, Varroa han mostrado resistencia al grupo de insecticidas piretroides, incluyendo Bayvarol ® que tiene la flumetrina como ingrediente activo. En el trabajo que aquí se presenta, se observaron cambios en los proteomas expresados por los ácaros, ya sea sensibles o resistentes a Bayvarol ® utilizando 2D-SDS-PAGE y proteó mica con marcaje libre shotgun. Ciertas proteínas de desintoxicació n (por ejemplo, transferasa-s-glutatió n, monooxigenasa incluyendo flavina) estaban presentes en niveles más altos en los ácaros resistentes, así como algunas proteínas de bombeo de protones (por ejemplo, las subunidades alfa y beta de la ATPasa Na + / K +, y la proteína ATPasa E1-E2). Se observó una disminució n en la abundancia de 12 proteínas de la cutícula de los ácaros resistentes lo que indica que la alteració n de la estructura de la cutícula podría ser un mecanismo de resistencia potencial. Ciertas proteínas estructurales, tales como la miosina y la alfa tubulina se expresaron en niveles más altos en los ácaros resistentes, lo que podrían indicar un cambio en la estructura intracelular de la barrera de la cutícula o un cambio en la form...

A Varroa destructor protein atlas reveals molecular underpinnings of developmental transitions and sexual differentiation

“…The work presented here provides a foundation to begin to unravel the fundamentals of Varroa biology, including developmental transitions, sexual differentiation, diet, and host–virus interactions, as well as assisting with improving the genome annotation. Prior to this, there has been one published Varroa RNA-seq transcriptomics ( 24 ) and two proteomics studies, one of which—as far as we can tell—identified only virus and honey bee proteins and none from Varroa ( 15 ), and the other only analyzed foundresses ( 26 ). With almost 20,000 unique peptides identified, our study represents the deepest Varroa proteome to date.…”

“…Research on Varroa has focused on its role as a vector for viruses ( 13 – 18 ), their response to pheromone cues ( 19 – 21 ), attempts to control it via RNAi ( 22 – 24 ), and host shifts ( 25 ). At the time of writing, there have only been two previous Varroa proteomic investigations, one of which focused on viral proteins ( 15 ) and the other identifying fewer than 700 proteins within one developmental stage ( 26 ). Global protein expression changes associated with developmental transitions and sexual differentiation are yet unknown.…”

Quantitative proteomics reveals divergent responses in Apis mellifera worker and drone pupae to parasitization by Varroa destructor

“…High-resolution quantitative mass spectrometry (MS) routinely quantify and identify thousands of proteins across multiple samples in a single run which gives an unprecedented opportunity to examine changes in proteomic profile of a particular biological fraction or organism. We previously utilised this technology to study the mechanisms conferring resistance to Bayvarol in V. destructor and the results indicated a decreased abundance of cuticle associated proteins and an increase in the abundance of proteins involved in cellular detoxification as possible factors in the resistant phenotype (Surlis et al, 2016). Similar proteome-wide analysis has been utilised to investigate the molecular determinants of hygienic behaviours and resistance/sensitivity to V. destructor (Parker et al, 2012;Hu et al, 2016).…”

AVarroa destructorprotein atlas reveals molecular underpinnings of developmental transitions and sexual differentiation

“…The work presented here provides a foundation to unravel the fundamentals of Varroa biology, including developmental transitions, sexual differentiation, diet, and host-virus interactions, as well as assisting with improving the genome annotation. Prior to this, there has been one published Varroa RNA-seq transcriptomics (24) and two proteomics studies, one of whichas far as we can tellidentified only virus and honey bee proteins and none from Varroa (15), and the other only analyzed foundresses (26). With almost 20,000 unique peptides identified, our study represents the deepest Varroa proteome to date.…”

“…Research on Varroa has focused on its role as a vector for viruses (13)(14)(15)(16)(17)(18), their response to pheromone cues (19-21), attempts to control it via RNAi (22)(23)(24) and host shifts (25). At the time of writing, there have only been two previous Varroa proteomic investigations, one of which focused on viral proteins (15) and the other identifying fewer than 700 proteins within one developmental stage (26). Global gene expression changes associated with developmental transitions and sexual differentiation are yet unknown.…”