Evaluating the Efficacy of 30 Different Essential Oils against Varroa destructor and Honey Bee Workers (Apis mellifera)

Essential oils and their components are generally known for their acaricidal effects and are used as an alternative to control the population of the Varroa destructor instead of synthetic acaricides. However, for many essential oils, the exact acaricidal effect against Varroa mites, as well as the effect against honey bees, is not known. In this study, 30 different essential oils were screened by using a glass-vial residual bioassay. Essential oils showing varroacidal efficacy > 70% were tested by the complete exposure assay. A total of five bees and five mites were placed in the Petri dishes in five replications for each concentration of essential oil. Mite and bee mortality rates were assessed after 4, 24, 48, and 72 h. The LC50 values and selectivity ratio (SR) were calculated. For essential oils with the best selectivity ratio, their main components were detected and quantified by GC-MS/MS. The results suggest that the most suitable oils are peppermint and manuka (SR > 9), followed by oregano, litsea (SR > 5), carrot, and cinnamon (SR > 4). Additionally, these oils showed a trend of the increased value of selective ratio over time. All these oils seem to be better than thymol (SR < 3.2), which is commonly used in beekeeping practice. However, the possible use of these essential oils has yet to be verified in beekeeping practice.

Effectiveness of Different Soft Acaricides against Honey Bee Ectoparasitic Mite Varroa destructor (Acari: Varroidae)

Honey bees (Apis mellifera) are essential for their products—honey, royal jelly, pollen, propolis and beeswax. They are also indispensable because they support ecosystems with their pollination services. However, the production and functions of honey bees are hindered by the arthropod pest Varroa destructor, which attacks bees through its feeding activities. Efforts to control varroa mites have been made through the development of various synthetic pesticide groups, but have had limited success because the mites developed resistance and some of these pesticides are harmful to bees. Branded pesticides are rarely used in Pakistan, as beekeepers utilize acaricides from unknown sources. There is a need to create awareness of available naturally occurring acaricides that may serve as an alternative to synthetic acaricides. Although some naturally occurring compounds are considered toxic to the environment, the soft acaricides oxalic acid, thymol, and formic acid 65% are usually safe for honey bee colonies and beekeepers, when handled appropriately. The current study investigated the effectiveness of formic acid (10, 15, and 20 mL/hive), oxalic acid (4.2, 3.2, and 2.1%/hive), and thymol (6, 4, and 2 g/hive) in controlling mite infestation. The results indicated that all treatments significantly reduced the mite population (p < 0.05). The average efficacies of oxalic acid at 3.2% (94.84% ± 0.34) and 4.2% (92.68% ± 0.37) were significantly higher than those of the other treatments. The lowest efficacy was recorded in formic acid 65% at 10 mL (54.13%). Overall, the results indicated that soft acaricides—such as oxalic acid at 3.2% and 4.2% concentrations—are very effective at controlling varroa mites and can be used in broodless conditions without side effects.

The Westernmost Mediterranean islands present an endemic varroosis disease pattern

Aim of study: This is the first intensive study of the incidence of Varroa destructor (Acari: Varroidae) in the Westernmost Mediterranean islands (Balearics). Area of study: Mallorca, Menorca, Eivissa and Formentera (Balearic Islands), Spain. Material and methods: The study was carried out in autumn 2010 and in spring 2011 because the presence of the mite is higher in these seasons. A total of 462 honeybee colonies were analyzed from different apiaries on. One sample of adult bees (worker bees, n > 200) and another of bee brood (at all stages of development, n> 400) were collected from each colony. All were frozen until analysis. To detect Varroa each sample was examined individually basing the study on the guide edited by the World Organisation for Animal Health, with some modifications. The beekeepers involved were interviewed to characterize Balearic beekeeping. Main results: Regarding the prevalence, results showed that in the Balearic Islands the percentage was higher in spring (34.78%) than in autumn (44.09%) but the infestation rate was lower in both seasons (Autumn: 0.94%, Spring: 0.77%). The focality percentage was also higher in spring (87.10%) than in autumn (82.61%). The data suggests that Varroa mites in the archipelago have an endemic rather than epidemic disease distribution pattern. Research highlights: These results open an interesting scenario to develop a future resistance breeding strategy against V. destructor in these small islands.

Residual Tau-Fluvalinate in Honey Bee Colonies Is Coupled with Evidence for Selection for Varroa destructor Resistance to Pyrethroids

Varroa destructor is considered one of the most devastating parasites of the honey bee, Apis mellifera, and a major problem for the beekeeping industry. Currently, the main method to control Varroa mites is the application of drugs that contain different acaricides as active ingredients. The pyrethroid tau-fluvalinate is one of the acaricides most widely used in beekeeping due to its efficacy and low toxicity to bees. However, the intensive and repetitive application of this compound produces a selective pressure that, when maintained over time, contributes to the emergence of resistant mites in the honey bee colonies, compromising the acaricidal treatments efficacy. Here we studied the presence of tau-fluvalinate residues in hives and the evolution of genetic resistance to this acaricide in Varroa mites from honey bee colonies that received no pyrethroid treatment in the previous four years. Our data revealed the widespread and persistent tau-fluvalinate contamination of beeswax and beebread in hives, an overall increase of the pyrethroid resistance allele frequency and a generalized excess of resistant mites relative to Hardy–Weinberg equilibrium expectations. These results suggest that tau-fluvalinate contamination in the hives may seriously compromise the efficacy of pyrethroid-based mite control methods.

An octopamine receptor confers selective toxicity of amitraz on honeybees and Varroa mites

The Varroa destructor mite is a devastating parasite of Apis mellifera honeybees. They can cause colonies to collapse by spreading viruses and feeding on the fat reserves of adults and larvae. Amitraz is used to control mites due to its low toxicity to bees; however, the mechanism of bee resistance to amitraz remains unknown. In this study, we found that amitraz and its major metabolite potently activated all four mite octopamine receptors. Behavioral assays using Drosophila null mutants of octopamine receptors identified one receptor subtype Octβ2R as the sole target of amitraz in vivo. We found that thermogenetic activation of octβ2R-expressing neurons mimics amitraz poisoning symptoms in target pests. We next confirmed that the mite Octβ2R was more sensitive to amitraz and its metabolite than the bee Octβ2R in pharmacological assays and transgenic flies. Furthermore, replacement of three bee-specific residues with the counterparts in the mite receptor increased amitraz sensitivity of the bee Octβ2R, indicating that relative insensitivity of their receptor is the major mechanism for honeybees to resist amitraz. The present findings have important implications for resistance management and the design of safer insecticides that selectively target pests while maintaining low toxicity to non-target pollinators.

Evaluating toxicity of Varroa mite (Varroa destructor)-active dsRNA to monarch butterfly (Danaus plexippus) larvae

Varroa mites (Varroa destructor) are parasitic mites that, combined with other factors, are contributing to high levels of honey bee (Apis mellifera) colony losses. A Varroa-active dsRNA was recently developed to control Varroa mites within honey bee brood cells. This dsRNA has 372 base pairs that are homologous to a sequence region within the Varroa mite calmodulin gene (cam). The Varroa-active dsRNA also shares a 21-base pair match with monarch butterfly (Danaus plexippus) calmodulin mRNA, raising the possibility of non-target effects if there is environmental exposure. We chronically exposed the entire monarch larval stage to common (Asclepias syriaca) and tropical (Asclepias curassavica) milkweed leaves treated with concentrations of Varroa-active dsRNA that are one- and ten-fold higher than those used to treat honey bee hives. This corresponded to concentrations of 0.025–0.041 and 0.211–0.282 mg/g leaf, respectively. Potassium arsenate and a previously designed monarch-active dsRNA with a 100% base pair match to the monarch v-ATPase A mRNA (leaf concentration was 0.020–0.034 mg/g) were used as positive controls. The Varroa mite and monarch-active dsRNA’s did not cause significant differences in larval mortality, larval or pupal development, pupal weights, or adult eclosion rates when compared to negative controls. Irrespective of control or dsRNA treatment, larvae that consumed approximately 7500 to 10,500-mg milkweed leaf within 10 to 12 days had the highest pupal weights. The lack of mortality and sublethal effects following dietary exposure to dsRNA with 21-base pair and 100% base pair match to mRNAs that correspond to regulatory genes suggest monarch mRNA may be refractory to silencing by dsRNA or monarch dsRNase may degrade dsRNA to a concentration that is insufficient to silence mRNA signaling.

A gene drive does not spread easily in populations of the honey bee parasite Varroa destructor

Varroa mites (Varroa destructor) are the most significant threat to beekeeping worldwide. They are directly or indirectly responsible for millions of colony losses each year. Beekeepers are somewhat able to control Varroa populations through the use of physical and chemical treatments. However, these methods range in effectiveness, can harm honey bees, can be physically demanding on the beekeeper, and do not always provide complete protection from Varroa. More importantly, in some populations Varroa mites have developed resistance to available acaricides. Overcoming the Varroa mite problem will require novel and targeted treatment options. Here, we explore the potential of gene drive technology to control Varroa. We show that spreading a neutral gene drive in Varroa is possible but requires specific colony-level management practices to overcome the challenges of both inbreeding and haplodiploidy. Furthermore, continued treatment with acaricides is necessary to give a gene drive time to fix in the Varroa population. Unfortunately, a gene drive that impacts female or male fertility does not spread in Varroa. Therefore, we suggest that the most promising way forward is to use a gene drive which carries a toxin precursor or removes acaricide resistance alleles.

Residual Tau-Fluvalinate in Honey Bee Colonies is Coupled with Evidence for Selection for Varroa Destructor Resistance to Pyrethroids

Varroa destructor is considered one of the most devastating parasites of the honey bee, Apis mellifera, and a major problem for the beekeeping industry. Currently, the main method to control Varroa mites is the application of drugs that contain different acaricides as active ingredients. The pyrethroid tau-fluvalinate is one of the acaricides most widely used in beekeeping due to its efficacy and low toxicity to bees. However, the intensive and repetitive application of this compound produces a selective pressure that, when maintained over time, contributes to the emergence of resistant mites in the honey bee colonies, compromising the acaricidal treatments efficacy. Here we studied the presence of tau-fluvalinate residues in hives and the evolution of genetic resistance to this acaricide in Varroa mites from honeybee colonies that received no pyrethroid treatment in the previous four years. Our data revealed the widespread and persistent tau-fluvalinate contamination of beeswax and beebread in hives, an overall increase of the pyrethroid resistance allele frequency and a generalized excess of resistant mites relative to Hardy-Weinberg equilibrium expectations. These results suggest that tau-fluvalinate contamination of the hives may seriously compromise the efficacy of pyrethroid-based mite control methods.

A New Strain of Virus Discovered in China Specific to the Parasitic Mite Varroa destructor Poses a Potential Threat to Honey Bees

The ectoparasitic mite, Varroa destructor, feeds directly on honey bees and serves as a vector for transmitting viruses among them. The Varroa mite causes relatively little damage to its natural host, the Eastern honey bee (Apis cerana) but it is the most devastating pest for the Western honey bee (Apis mellifera). Using Illumina HiSeq sequencing technology, we conducted a metatranscriptome analysis of the microbial community associated with Varroa mites. This study led to the identification of a new Chinese strain of Varroa destructor virus-2 (VDV-2), which is a member of the Iflaviridae family and was previously reported to be specific to Varroa mites. A subsequent epidemiological investigation of Chinese strain of VDV-2 (VDV-2-China) showed that the virus was highly prevalent among Varroa populations and was not identified in any of the adult workers from both A. mellifera and A.cerana colonies distributed in six provinces in China, clearly indicating that VDV-2-China is predominantly a Varroa-adapted virus. While A. mellifera worker pupae exposed to less than two Varroa mites tested negative for VDV-2-China, VDV-2-China was detected in 12.5% of the A. mellifera worker pupae that were parasitized by more than 10 Varroa mites, bringing into play the possibility of a new scenario where VDV-2 could be transmitted to the honey bees during heavy Varroa infestations. Bioassay for the VDV-2-China infectivity showed that A. cerana was not a permissive host for VDV-2-China, yet A. mellifera could be a biological host that supports VDV-2-China’s replication. The different replication dynamics of the virus between the two host species reflect their variation in terms of susceptibility to the virus infection, posing a potential threat to the health of the Western honey bee. The information gained from this study contributes to the knowledge concerning genetic variabilities and evolutionary dynamics of Varroa-borne viruses, thereby enhancing our understanding of underlying molecular mechanisms governing honey bee Varroosis.