Mechanisms of Pathogen and Pesticide Resistance in Honey Bees.

Bees are the most important insect pollinators of the crops humans grow, and Apis mellifera, the Western honey bee, is the most commonly managed species for this purpose. In addition to providing agricultural services, the complex biology of honey bees has been the subject of scientific study since the 18th century, and the intricate behaviors of honey bees and ants, fellow hymenopterans, inspired much sociobiological inquest. Unfortunately, honey bees are constantly exposed to parasites, pathogens, and xenobiotics, all of which pose threats to their health. Despite our curiosity about and dependence on honey bees, defining the molecular mechanisms underlying their interactions with biotic and abiotic stressors has been challenging. The very aspects of their physiology and behavior that make them so important to agriculture also make them challenging to study, relative to canonical model organisms. However, because we rely on A. mellifera so much for pollination, we must continue our efforts to understand what ails them. Here, we review major advancements in our knowledge of honey bee physiology, focusing on immunity and detoxification, and highlight some challenges that remain.

Stress-induced loss of social resilience in honeybee colonies and its implications on fitness.

Stressors may lead to a shift in the timing of life-history events of species, causing a mismatch with optimal environmental conditions, potentially reducing fitness. In honeybees, the timing of brood rearing and nest emergence in late winter/early spring is critical as colonies need to grow fast after winter to prepare for reproduction. However, the effects of stress on these life-history events in late winter/early spring and the possible consequences are not well understood. Therefore, we tested whether (i) honeybee colonies shift timing of brood rearing and nest emergence as response to stressors, and (ii) if there is a consequent loss of social resilience, reflected in colony fitness (survival, growth and reproduction). We monitored stressed (high load of the parasitic mite Varroa destructor or nutrition restricted) colonies and presumably non-stressed colonies from the beginning of 2020 till spring of 2021. We found that honeybee colonies do not shift the timing of brood rearing and nest emergence in spring as a coping mechanism to stressors. However, we show that there is loss of social resilience in stressed colonies, leading to reduced growth and reproduction. Our study contributes to better understanding the effects of stressors on social resilience in eusocial organisms.

Functional redundancy and niche specialization in honeybee and Varroa microbiomes.

The honeybee (Apis mellifera) is a key pollinator critical to global agriculture, facing threats from various stressors, including the ectoparasitic Varroa mite (Varroa destructor). Previous studies have identified shared bacteria between Varroa mites and honeybees, yet it remains unclear if these bacteria assemble similarly in both species. This study builds on existing knowledge by investigating co-occurrence patterns in the microbiomes of both Varroa mites and honeybees, shedding light on potential interactions. Leveraging 16S rRNA datasets, we conducted co-occurrence network analyses, explored Core Association Networks (CAN) and assess network robustness. Comparative network analyses revealed structural differences between honeybee and mite microbiomes, along with shared core features and microbial motifs. The mite network exhibited lower robustness, suggesting less resistance to taxa extension compared to honeybees. Furthermore, analyses of predicted functional profiling and taxa contribution revealed that common central pathways in the metabolic networks have different taxa contributing to Varroa mites and honeybee microbiomes. The results show that while both microbial systems exhibit functional redundancy, in which different taxa contribute to the functional stability and resilience of the ecosystem, there is evidence for niche specialization resulting in unique contributions to specific pathways in each part of this host-parasite system. The specificity of taxa contribution to key pathways offers targeted approaches to Varroa microbiome management and preserving honeybee microbiome. Our findings provide valuable insights into microbial interactions, aiding farmers and beekeepers in maintaining healthy and resilient bee colonies amid increasing Varroa mite infestations.

Relative impacts of Varroa destructor (Mesostigmata:Varroidae) infestation and pesticide exposure on honey bee colony health and survival in a high-intensity corn and soybean producing region in northern Iowa.

The negative effects of Varroa and pesticides on colony health and survival are among the most important concerns to beekeepers. To compare the relative contribution of Varroa, pesticides, and interactions between them on honey bee colony performance and survival, a 2-year longitudinal study was performed in corn and soybean growing areas of Iowa. Varroa infestation and pesticide content in stored pollen were measured from 3 apiaries across a gradient of corn and soybean production areas and compared to measurements of colony health and survival. Colonies were not treated for Varroa the first year, but were treated the second year, leading to reduced Varroa infestation that was associated with larger honey bee populations, increased honey production, and higher colony survival. Pesticide detections were highest in areas with high-intensity corn and soybean production treated with conventional methods. Pesticide detections were positively associated with honey bee population size in May 2015 in the intermediate conventional (IC) and intermediate organic (IO) apiaries. Varroa populations across all apiaries in October 2015 were negatively correlated with miticide and chlorpyrifos detections. Miticide detections across all apiaries and neonicotinoid detections in the IC apiary in May 2015 were higher in colonies that survived. In July 2015, colony survival was positively associated with total pesticide detections in all apiaries and chlorpyrifos exposure in the IC and high conventional (HC) apiaries. This research suggests that Varroa are a major cause of reduced colony performance and increased colony losses, and honey bees are resilient upon low to moderate pesticide detections.

Current honey bee stressor investigations and mitigation methods in the United States and Canada.

Honey bees are the most important managed insect pollinators in the US and Canadian crop systems. However, the annual mortality of colonies in the past 15 years has been consistently higher than historical records. Because they are eusocial generalist pollinators and amenable to management, honey bees provide a unique opportunity to investigate a wide range of questions at molecular, organismal, and ecological scales. Here, the American Association of Professional Apiculturists (AAPA) and the Canadian Association of Professional Apiculturists (CAPA) created 2 collections of articles featuring investigations on micro and macro aspects of honey bee health, sociobiology, and management showcasing new applied research from diverse groups studying honey bees (Apis mellifera) in the United States and Canada. Research presented in this special issue includes examinations of abiotic and biotic stressors of honey bees, and evaluations and introductions of various stress mitigation measures that may be valuable to both scientists and the beekeeping community. These investigations from throughout the United States and Canada showcase the wide breadth of current work done and point out areas that need further research.

Evaluating the seasonal efficacy of commonly used chemical treatments on Varroa destructor (Mesostigmata: Varroidae) population resurgence in honey bee colonies.

The purpose of this research was to determine how common chemical treatments influence Varroa destructor (Anderson and Trueman) population resurgence rates (defined as time posttreatment for mite populations to reach 3 mites/100 adult bees) in managed honey bee (Apis mellifera L.) colonies seasonally. We conducted 2 experiments that followed the same basic protocol to address this purpose. We established 6 treatment groups in Experiment 1 in the fall of 2014: untreated control, Apivar, Apistan, CheckMite+, ApiLifeVar, and Mite Away II applied to 10 colonies per treatment. In Experiment 2, we applied 8 chemical treatments to each of 4 seasonal (spring, summer, fall, and winter) cohorts of honey bee colonies to determine how mite populations are influenced by the treatments. The treatments/formulations tested were Apivar, Apistan, Apiguard, MAQS, CheckMite+, oxalic acid (dribble), oxalic acid (shop towels), and amitraz (shop towels soaked in Bovitraz). In Experiment 1, Apivar and Mite Away II were able to delay V. destructor resurgence for 2 and 6 months, respectively. In Experiment 2, Apiguard, MAQS, oxalic acid (dribble), and Bovitraz treatments were effective at delaying V. destructor resurgence for at least 2 months during winter and spring. Only the Bovitraz and MAQS treatments were effective at controlling V. destructor in the summer and fall. Of the 2 amitraz-based treatments, the off-label Bovitraz treatment was the only treatment to reduce V. destructor populations in every season. The data gathered through this study allow for the refinement of treatment recommendations for V. destructor, especially regarding the seasonal efficacy of each miticide and the temporal efficacy posttreatment.

Sensorizing a Beehive: A Study on Potential Embedded Solutions for Internal Contactless Monitoring of Bees Activity.

Winter is the season of main concern for beekeepers since the temperature, humidity, and potential infection from mites and other diseases may lead the colony to death. As a consequence, beekeepers perform invasive checks on the colonies, exposing them to further harm. This paper proposes a novel design of an instrumented beehive involving color cameras placed inside the beehive and at the bottom of it, paving the way for new frontiers in beehive monitoring. The overall acquisition system is described focusing on design choices towards an effective solution for internal, contactless, and stress-free beehive monitoring. To validate our approach, we conducted an experimental campaign in 2023 and analyzed the collected images with YOLOv8 to understand if the proposed solution can be useful for beekeepers and what kind of information can be derived from this kind of monitoring, including the presence of Varroa destructor mites inside the beehive. We experimentally found that the observation point inside the beehive is the most challenging due to the frequent movements of the bees and the difficulties related to obtaining in-focus images. However, from these images, it is possible to find Varroa destructor mites. On the other hand, the observation point at the bottom of the beehive showed great potential for understanding the overall activity of the colony.

Geographical distribution of pyrethroid resistance mutations in Varroa destructor across Türkiye and a European overview.

Varroa destructor Anderson & Trueman (Acari: Varroidae) is of paramount significance in modern beekeeping, with infestations presenting a primary challenge that directly influences colony health, productivity, and overall apicultural sustainability. In order to control this mite, many beekeepers rely on a limited number of approved synthetic acaricides, including the pyrethroids tau-fluvalinate, flumethrin and organophosphate coumaphos. However, the excessive use of these substances has led to the widespread development of resistance in various beekeeping areas globally. In the present study, the occurrence of resistance mutations in the voltage-gated sodium channel (VGSC) and acetylcholinesterase (AChE), the target-site of pyrethroids and coumaphos, respectively, was examined in Varroa populations collected throughout the southeastern and eastern Anatolia regions of Türkiye. All Varroa samples belonged to the Korean haplotype, and a very low genetic distance was observed based on cytochrome c oxidase subunit I (COI) gene sequences. No amino acid substitutions were determined at the key residues of AChE. On the other hand, three amino acid substitutions, (L925V/I/M), previously associated with pyrethroid resistance, were identified in nearly 80% of the Turkish populations. Importantly, L925M, the dominant mutation in the USA, was detected in Turkish Varroa populations for the first time. To gain a more comprehensive perspective, we conducted a systematic analysis of the distribution of pyrethroid resistance mutations across Europe, based on the previously reported data. Varroa populations from Mediterranean countries such as Türkiye, Spain, and Greece exhibited the highest frequency of resistance mutation. Revealing the occurrence and geographical distribution of pyrethroid resistance mutations in V. destructor populations across the country will enhance the development of more efficient strategies for mite management.

Developing a method to rear Varroa destructor in vitro.

Varroa destructor is a significant mite pest of western honey bees (Apis mellifera). Developing a method to rear and maintain populations of V. destructor in vitro would provide year-round access to the mites, allowing scientists to study their biology, behavior, and control more rapidly. In this study, we determined the impact of various rearing parameters on V. destructor survival and reproduction in vitro. This was done by collecting V. destructor from colonies, placing them in gelatin capsules containing honey bee larvae, and manipulating the following conditions experimentally: rearing temperature, colony source of honey bee larva, behavioral/developmental stages of V. destructor and honey bee larva, and mite:bee larva ratio. Varroa destructor survival was significantly impacted by temperature, colony source of larvae and mite behavioral stage. In addition, V. destructor reproduction was significantly impacted by mite: larva ratio, larval developmental stage, colony source of larva, and temperature. The following conditions optimized mite survival and reproduction in vitro: using a 4:1 mite:larva ratio, beginning the study with late stage uncapped larvae, using mites collected from adult bees, maintaining the rearing temperature at 34.5° C, and screening larval colony source. Ultimately, this research can be used to improve V. destructor in vitro rearing programs.

Promising Algerian essential oils as natural acaricides against the honey bee mite Varroa destructor (Acari: Varroidae).

Varroosis induced by Varroa destructor Anderson and Trueman represents the most pathogenic and destructive disease affecting the western honey bee, Apis mellifera. In this study, we investigated the acaricidal activity against the Varroa mite using essential oils (EOs) from the aerial parts of four autochthonous Algerian herbal species, namely Artemisia herba alba, Artemisia campestris, Artemisia judaica and Ruta montana. EOs were obtained by means of hydrodistillation and their composition was characterized by gas chromatography-mass spectrometry. The toxicity of the selected EOs toward V. destructor and A. mellifera adult honey bees was evaluated using the complete exposure method. The results indicate the predominance of davanone (66.9%) in A. herba alba, ß-pinene (19.5%) in A. campestris, piperitone (68.7%) in A. judaica and 2-undecanone (70.1%) in R. montana EOs. Interestingly, the LC50 values coupled to bee mortality rates revealed that all tested oils exhibited significant acaricidal efficiency with selectivity ratio (SR) values of 10.77, 8.78, 5.62 and 3.73 for A. campestris, A. judaica, A. herba alba, and R. montana, respectively. These values were better than that of thymol (SR = 3.65), the positive control. These findings suggest that these EOs could be used as plant-derived veterinary acaricides to control varroosis in field conditions.

Signatures of adaptive decreased virulence of deformed wing virus in an isolated population of wild honeybees (Apis mellifera).

Understanding the ecological and evolutionary processes that drive host-pathogen interactions is critical for combating epidemics and conserving species. The Varroa destructor mite and deformed wing virus (DWV) are two synergistic threats to Western honeybee (Apis mellifera) populations across the globe. Distinct honeybee populations have been found to self-sustain despite Varroa infestations, including colonies within the Arnot Forest outside Ithaca, NY, USA. We hypothesized that in these bee populations, DWV has been selected to produce an avirulent infection phenotype, allowing for the persistence of both host and disease-causing agents. To investigate this, we assessed the titre of viruses in bees from the Arnot Forest and managed apiaries, and assessed genomic variation and virulence differences between DWV isolates. Across groups, we found viral abundance was similar, but DWV genotypes were distinct. We also found that infections with isolates from the Arnot Forest resulted in higher survival and lower rates of symptomatic deformed wings, compared to analogous isolates from managed colonies, providing preliminary evidence to support the hypothesis of adaptive decreased viral virulence. Overall, this multi-level investigation of virus genotype and phenotype indicates that host ecological context can be a significant driver of viral evolution and host-pathogen interactions in honeybees.

Frequent Parasitism of Apis mellifera by Trypanosomatids in Geographically Isolated Areas with Restricted Beekeeping Movements.

Trypanosomatids form a group of high prevalence protozoa that parasitise honey bees, with Lotmaria passim as the predominant species worldwide. However, the knowledge about the ecology of trypanosomatids in isolated areas is limited. The Portuguese archipelagos of Madeira and Azores provide an interesting setting to investigate these parasites because of their geographic isolation, and because they harbour honey bee populations devoid of two major enemies: Varroa destructor and Nosema ceranae. Hence, a total of 661 honey bee colonies from Madeira and the Azores were analysed using different molecular techniques, through which we found a high prevalence of trypanosomatids despite the isolation of these islands. L. passim was the predominant species and, in most colonies, was the only one found, even on islands free of V. destructor and/or N. ceranae with severe restrictions on colony movements to prevent the spread of them. However, islands with V. destructor had a significantly higher prevalence of L. passim and, conversely, islands with N. ceranae did not shown any significant correlation with the trypanosomatid. Crithidia bombi was detected in Madeira and on three islands of the Azores, almost always coincident with L. passim. By contrast, Crithidia mellificae was not detected in any sample. A high-throughput sequencing analysis distinguished two main haplotypes of L. passim, which accounted for 98% of the total sequence reads. This work suggests that L. passim and C. bombi are parasites that have been associated with honey bees predating the spread of V. destructor and N. ceranae.

Short communication: Varroa destructor re-invasion dynamics during autumn and winter in Apis mellifera colonies from a temperate climate.

Infestation with Varroa destructor compromises the survival of Apis mellifera colonies, especially during the winter season. In order to prevent colony losses due to the presence of Varroa mites, determination of the infestation level and the application of autumn treatment is highly recommended. However, autumn reinvasion by inter-apiary and intra-apiary horizontal transmission might threaten the control strategies. Drivers like the infestation level of untreated colonies, the presence of bee brood during late autumn and the colonies disposition within the apiary might explain at least partially the re-invasion rates of treated colonies during autumn and winter. A total of six apiaries with five colonies were distributed within a 30 km radius. Colonies were arranged within each apiary following a circular or a lineal disposition. Twenty-four of the thirty colonies were treated during late summer against V. destructor and one colony per apiary remained untreated. The phoretic Varroa infestation and the adult bee population, brood area and nutritional reserves in the colonies was estimated before and after treatment and then for the next four months (late autumn and winter season). In this study, a model for the V. destructor re-invasion dynamics during autumn and winter in honey bee colonies from a temperate climate was fitted. According to these results, the re-invasion probability is higher for lineal disposition of the colonies that also present a higher amount of brood availability during late autumn and winter. Beekeeping practical implications are discussed in the context of an Integrated Pest management approach.

Lithium chloride treatments in free flying honey bee colonies: efficacy, brood survival, and within-colony distribution.

The efficacy of various lithium chloride (LiCl) applications in eradicating the parasitic mite Varroa destructor in honey bee colonies was investigated, with a specific focus on its impact on brood development. In broodless colonies (3 weeks post queen caging), the highest efficacy of 98% was achieved with a 9-day treatment of 2.5 kg of candy spiked with 50 mM LiCl. A shorter 5-day treatment with 2 kg of 50 mM LiCl candy resulted in an efficacy of 78%. In colonies with brood, a repeated short-term application of 4 × 0.5 kg 50 mM LiCl candy yielded an efficacy of 88%. LiCl treatment led to a removal of the first batch of brood reared after release of the queen. However, no long-term effects on colony growth were observed, and the colonies successfully overwintered. Additionally, the study demonstrated that lithium is rapidly distributed among the bees of a colony within 2 days, yet only low concentrations were detected in stored food samples. This suggests that the bees efficiently absorb and distribute lithium within the colony. The harvested honey in the following spring revealed a lithium concentration of 0.1-0.2 mg/kg, which is below naturally occurring lithium levels in honey. Based on these findings, LiCl can be considered an effective and easy-to-apply acaricide in broodless colonies, and even in colonies with brood, it had good efficacy and no long-term effects on colony survival. Further research may be necessary to determine the optimal treatment period for achieving an efficacy over 95%.

Oxalic acid application method and treatment intervals for reduction of Varroa destructor (Mesostigmata: Varroidae) populations in Apis mellifera (Hymenoptera: Apidae) colonies.

Oxalic acid (OA) is a popular miticide used to control Varroa destructor (Mesostigmata: Varroidae) in western honey bee (Apis mellifera L.) (Hymenoptera: Apidae) colonies. Our aim was to investigate which method of OA application (dribbling, fogging, or vaporizing) was the most effective at reducing V. destructor infestations (Experiment 1) and to improve upon this method by determining the treatment interval that resulted in the greatest V. destructor control (Experiment 2). We used the product Api-Bioxal (97% OA) and maintained 40 honey bee colonies (10/treatment) in both experiments. In Experiment 1, the treatments included (i) dribbling 50 ml of 3% OA solution, (ii) vaporizing 4 g of solid OA, (iii) using an insect fogger supplied with 2.5% OA dissolved in ethyl alcohol, and (iv) an untreated control. After 3 weeks, only the vaporization method reduced V. destructor infestations (from 9.24 mites/100 bees pretreatment to 3.25 mites/100 bees posttreatment) and resulted in significantly increased brood amounts and numbers of adult bees over those of the controls. In Experiment 2, all colonies were treated with 4 applications of OA via vaporization at a constant concentration of 4 g OA/colony. In this experiment, the groups were separated by treatment intervals at either 3-, 5-, or 7-day intervals. We observed that 5- and 7-day treatment intervals significantly reduced V. destructor populations from pretreatment levels over that of the controls and 3-day intervals. Our data demonstrate the efficacy of OA in reducing V. destructor infestation, particularly vaporizing 4 g every 5-7 days as the most effective method of application.

Genetic variability of the honey bee mite, Varroa destructor, from a humid continental climatic region of Canada, and temperate and tropical climatic regions of Mexico.

Varroa destructor is a damaging mite of Western honey bees (Apis mellifera). Genetic variability of the mite in different regions of the world could be related to the movement of infested bees or other factors, such as climate. In this study, V. destructor samples were collected from tropical and temperate climate regions of Mexico, and a humid continental climate region of Canada. COX-1 AFLPs showed that all the mites were the Korean haplotype. Four microsatellites revealed nine haplogroups from the continental climate region of Canada, compared to three haplogroups from the tropical and temperate climate regions of Mexico. CytII-ATP sequences showed seven haplogroups from the humid continental climate region vs. two haplogroups from the temperate region and one haplogroup from the tropical region. CytB sequences revealed seven haplogroups from Canada vs. three from Mexico. A comparison of the cytB sequences of the samples from Canada and Mexico to those from a worldwide collection showed that one sequence, designated the cytB1 type, predominated, comprising 57% of the 86 sequences; it clustered with similar sequences that comprised 80% of the sequences, designated family A. CytB1 was predominant in Mexico, but not in Canada. The other 20% of sequences were in families B and C, and all those samples originated from East and Southeast Asia. The microsatellite, cytII-ATP, and cytB markers, all showed higher variability in mites collected in Canada than in Mexico, which could be related to the cooler climate or an earlier invasion and/or multiple mite invasions in Canada.

Cold case: The disappearance of Egypt bee virus, a fourth distinct master strain of deformed wing virus linked to honeybee mortality in 1970's Egypt.

In 1977, a sample of diseased adult honeybees (Apis mellifera) from Egypt was found to contain large amounts of a previously unknown virus, Egypt bee virus, which was subsequently shown to be serologically related to deformed wing virus (DWV). By sequencing the original isolate, we demonstrate that Egypt bee virus is in fact a fourth unique, major variant of DWV (DWV-D): more closely related to DWV-C than to either DWV-A or DWV-B. DWV-A and DWV-B are the most common DWV variants worldwide due to their close relationship and transmission by Varroa destructor. However, we could not find any trace of DWV-D in several hundred RNA sequencing libraries from a worldwide selection of honeybee, varroa and bumblebee samples. This means that DWV-D has either become extinct, been replaced by other DWV variants better adapted to varroa-mediated transmission, or persists only in a narrow geographic or host range, isolated from common bee and beekeeping trade routes.

Lethal concentrations of Cymbopogon nardus essential oils and their main component citronellal on Varroa destructor and Apis mellifera.

Varroosis is a disease caused by the mite Varroa destructor, and it is considered one of the biggest threats to honey bee populations globally. Mite control is centered on the use of synthetic acaricides, such as amitraz and flumethrine. However, high usage of these chemicals is associated with a wide variety of undesirable effects on bee colonies, including the development of resistance and persistence of harmful residues of acaricides in hive products used by humans. Botanical extracts have been identified as a potentially suitable organic alternative to synthetic acaricides. Essential oils, such as clove, eucalyptus, lemongrass, and oregano, have been found to exhibit acaricidal activity against V. destructor. The main goal of this work was to assess the bioactivity of the Cymbopogon nardus essential oil from two different locations (Argentina and India), and the activity of its major component the monoterpene citronellal. According to our results, complete essential oil from India is more effective in controlling parasitosis than the isolated citronellal component. The essential oil of C. nardus from Argentina demonstrated promise for the control of varroosis, as well as exhibiting low toxicity against bees (LC50 = 11.84 µL/mL). In addition, this essential oil may avoid the problems caused by synthetic acaricides, such as the emergence of resistance foci in Varroa and residues in hive products. Future research needs to investigate the delivery of volatile essentials oils to target mite populations.

Molecular and kinetic properties of three acetylcholinesterases in the Varroa mite, Varroa destructor.

The Varroa mite, Varroa destructor, poses one of the most serious threats to honey bees worldwide. Although coumaphos, an anticholinesterase pesticide, is widely used for varroa mite control, little information is available on the properties of Varroa mite acetylcholinesterases (VdAChEs). In this study, three putative VdAChEs were annotated and named VdAChE1, VdAChE2, and VdAChE3. All VdAChEs possessed most of the functionally important signature domains, suggesting that they are catalytically active. Phylogenetic analysis revealed that VdAChE1 was clustered into a clade containing most arthropod AChE1s, whereas VdAChE2 and VdAChE3 formed a unique clade with other arachnid AChEs. VdAChE1 was determined to be membrane-anchored, but both VdAChE2 and VdAChE3 are soluble, as judged by electrophoresis in conjunction with western blotting. Tissue-specific transcription profiling revealed that VdAChE1 was most predominantly expressed in the synganglion. In contrast, VdAChE2 was most predominantly expressed in the legs and cuticle. VdAChE3 showed negligible expression levels in all the tissues examined. In a kinetic analysis using recombinant VdAChEs, VdAChE1 exhibited the highest catalytic efficiency, followed by VdAChE2 and VdAChE3. Inhibition experiments revealed that VdAChE1 was most sensitive to all tested inhibitors. Taken together, VdAChE1 appears to be the major synaptic enzyme with a more toxicological relevance, whereas VdAChE2 is involved in other noncatalytic functions, including chemical defense against xenobiotics. Current findings contribute to a more detailed understanding of the evolutionary and functional traits of VdAChEs and to the design of novel anticholinesterase varroacides.

Genome-wide identification of miRNAs and target regulatory network in the invasive ectoparasitic mite Varroa destructor.

Varroa destructor is an ectoparasite mite that attacks bees leading to colony disorders worldwide. microRNAs (miRNAs) are key molecules used by eukaryotes to post-transcriptional control of gene expression. Nevertheless, still lack information aboutV. destructor miRNAs and its regulatory networks. Here, we used an integrative strategy to characterize the miRNAs in the V. destructor mite. We identified 310 precursors that give rise to 500 mature miRNAs, which 257 are likely mite-specific elements. miRNAs showed canonical length ranging between 18 and 25 nucleotides and 5' uracil preference. Top 10 elements concentrated over 80% of total miRNA expression, with bantam alone representing ~50%. We also detected non-templated bases in precursor-derived small RNAs, indicative of miRNA post-transcriptional regulatory mechanisms. Finally, we note that conserved miRNAs control similar processes in different organisms, suggesting a conservative role. Altogether, our findings contribute to the better understanding of the mite biology that can assist future studies on varroosis control.