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The Varroa destructor represents a significant threat to honey bees, leading to substantial yield losses and declines in colony health. Defense behaviors, including grooming (auto and allogrooming), serve as crucial mechanisms against Varroa infestations, yet the genetic basis of these behaviors remains elusive. This study examines the expression levels of hymenoptaecin (hym), neurexin-1 (AmNrx1), and CYP9Q3, potentially associated with defense behavior, in colonies of the Muğla honey bee ecotype (Apis mellifera anatoliaca) subjected to a Varroa selection program. Worker bees from 23 control groups and 23 colonies under selection were evaluated by using qPCR analysis. Results reveal a significant upregulation of hym, AmNrx1, and CYP9Q3 genes in the selected group, with fold changes of 2.9, 2.95, and 3.26 respectively compared to controls (p < 0.01). This suggests that selection against Varroa induces alterations in gene expression linked to Varroa exposure behaviors. These findings advocate for the potential use of hym, AmNrx1, and CYP9Q3 genes in preselection for future Varroa-resistant programs in honey bees. Supported by previous studies, these genes may facilitate the establishment of populations with enhanced defense behaviors, such as autogrooming and allogrooming., Competing Interests: Declarations Consent for publication All authors have seen and approved the final version of the manuscript being submitted. They warrant that the article is the authors’ original work, has not received prior publication, and is not under consideration for publication elsewhere. Ethics committee aproval As non cepholopad invertabrates honey bee not subject to animal ethic approval at Republic of Turkey (ased on the ‘Regulation on the Welfare and protection of Animal Used for Exsperimental and Other Scientific Purpose’ published in the official Gazette dated December 13.2011 and numbered 28141). Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Lactic acid is an alternative treatment to hard chemicals against Varroa destructor, the parasitic mite of the Western honey bee Apis mellifera. This soft acaricide is used only for small apiaries due to its laborious administration. However, the mode of action of this honey bee medication remains unknown. Previous studies showed that a direct contact between the arolia of V. destructor and lactic acid altered their morphology and led to an impairment of grip. Yet, there is no evidence for the way of action of lactic acid in a realistic in-hive scenario, i.e. after an indirect exposure of the mite through honey bees. We investigated the nature of lactic acid activity in the hive treatment context. The local and/or systemic way of action of this honey bee treatment against V. destructor was studied through a behavioural and toxicological approach at the individual level. On one hand, we confirmed the altered morphology for the arolia of mites and studied the evolution of the process over time. On the other hand, we found that haemolymph contaminated with lactic acid did not kill the feeding parasitic mite. These findings support a local mode of action. In order to unravel the sequence of events leading to the local contact between the acid and the mite on bees, we also documented the olfactory valence of lactic acid for A. mellifera and V. destructor. This work provides a new comprehension of lactic acid activity against the parasitic mite through honey bee exposure and gives new opportunities for control strategies against V. destructor., (© 2024. The Author(s).)

The growing demand for honey bee pollination services induces fluctuations in their populations and changes in bee health status. The current study aims to characterize the health condition of managed honey bees in three territories of the Department of Lambayeque, Peru, from a holistic perspective, considering several factors affecting bee health, including management practices, traits of the beekeepers and their apiaries, colony strength, infestation rates by Varroa sp., and the presence of agrochemical residues in bee bread. The results showed changes in land use, with large areas of crops dependent on bee pollination, determining their exposition to agrochemical residues. The 67% of bee bread samples presented pesticide residues, mainly, chlorpyrifos, carbendazim, boscalid, and azoxystrobin. In addition, limited expertise on good management practices by the beekeepers (i.e., inadequate disinfection of beekeeping materials, unsatisfactory varroa mites' surveillance, lack of protein supplements, presence of sanitary gaps) leads to deficiencies in bee colony strength and a high Varroa sp. prevalence in the territory, altering the bee health condition. To the best of our knowledge, this article illustrates the impact of the strongly anthropized ecosystems found along Peru's coast's arid zone on managed populations of honey bees in that territory with a One Health perspective.

Honey bees and other pollinators face threats from pesticides, imperfect nutrition, and a diverse set of parasites and pathogens. Honey bees are also a research model for development, social behavior, microbiology, and aging. Tackling these questions requires a mix of in-hive and controlled laboratory experiments. We have perfected small-scale, inexpensive, disposable, and rearing arenas for honey bees that have proved useful for hundreds of bioassays with thousands of bees. We describe those arenas here, show their advantages over current hoarding cages, and provide videos demonstrating their many uses., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

The honeybee (Apis mellifera) ectoparasite, Varroa destructor, is one of the most important honeybee pests worldwide. Acaricides, including the pyrethroids (tau-fluvalinate, flumethrin), and organophosphate (coumaphos) have been applied to control this mite within apiaries, still the long-term, constant, and excessive use of these products has led to the development of resistance in many populations. Three different mutations (L925V, L925I, L925M) at position 925 and one mutation (M918L) at 918 position of the V. destructor voltage-gated sodium channel (VGSC) have been associated with the resistance to these compounds. In the present study, we examined the presence of resistance mutations in the VGSC gene, encoding the target of pyrethroids, in the V. destructor population collected from the Kayseri and Sivas provinces of Central Anatolia Region of Türkiye. A total of 200 V. destructor samples were collected from 20 apiaries in two provinces throughout 2023. To investigate the mutations in Varroa samples, the domain II region of the VGSC gene was amplified using PCR and sequenced. The nucleotide sequencing of the IIS4-IIS5 linker region of the VGSC gene revealed one amino acid change at position 925: a leucine to isoleucine substitution (L925I). No mutations at other positions were identified. Homozygous resistant alleles were detected in 20 (40 %) of the sequenced 50 samples in the study areas. However, we detected the homozygous sensitive allele (wild-type allele, L925) in the remaining samples (50/30, 60 %). The result shows that this status may indicate a problem for Varroa control in the future. Thus, alternative acaricide with a mode of action of different pyrethroids should be considered in the control of V. destructor populations in these provinces., Competing Interests: Declaration of competing interest On behalf of all authors, the corresponding author states that there is no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Division of labour (DOL) in eusocial insects plays an important role in colony fitness. Honeybees face a variety of stressors that compromise the homeostasis of the colony and reduce survival and reproduction. Considering the significance of DOL in colony homeostasis, it is important to understand whether and how DOL may be altered as a result of chronic stress. Therefore, we tested whether honeybee colonies shift DOL in response to high infestation with the parasitic mite Varroa destructor. For this, we monitored chronically stressed and presumably low-stress colonies from April till December 2022. During the experiment, we applied a cold shock to test whether a perturbation resulted in a larger alteration in DOL in chronically stressed colonies. We found that after cold shock, there was a lower proportion of nurses in the chronically stressed colonies. For foragers, we found higher activity post-cold shock in chronically stressed colonies, but no difference between treatments in nectar inflow, suggesting less efficient foragers. Furthermore, we found that there was an accelerated task switch in chronically stressed colonies after the cold shock. The large changes after the perturbation may indicate inefficient task allocation due to chronic stress. Our study contributes to the understanding of social resilience and chronic stress responses in eusocial animals., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

The greater wax moth Galleria mellonella is a cosmopolitan pest of hives of the western honey bee Apis mellifera, where it remains exposed to varroicides applied by beekeepers in past decades as pest management chemicals for control of Varroa destructor, a devastating ectoparasite of bees. The prolonged presence of coumaphos residues, an organophosphate varroicide, in beeswax may be responsible for current levels of tolerance exhibited by G. mellonella, a non-target species that infests beehives. In this study, a field-collected strain of waxworms exhibited a higher LC 50 value for coumaphos than that of a laboratory strain that had not been continuously exposed to coumaphos residues at field concentrations. Despite its higher tolerance for coumaphos, the field strain experienced growth inhibition at ecologically relevant concentration of coumaphos. Moreover, at low environmental concentrations that did not alter growth, detoxification gene expression levels were substantially altered. RNA-Seq analysis revealed 1181 and 658 differentially expressed genes in fat body and midgut, respectively, with 378 and 186 of those genes upregulated. This large-scale upregulation encompassed 21 genes encoding cytochrome P450 monooxygenases (CYPs), 13 encoding UDP-glycosyltransferases (UGTs), 5 encoding glutathione-S-transferases (GSTs), 2 encoding carboxylesterases (COEs), and 2 encoding ABC transporters (ABCs) in either tissue. Expression analysis of 13 selected candidate detoxification genes by RT-qPCR was consistent with their expression from RNA-Seq data. In sum, our results indicate that long-lasting pesticide residues in beeswax from past Varroa mite management may continue to act as selective agents on detoxification systems of hive residents other than the initial target species and that multiple resistance mechanisms to these chemicals may coexist within the beehive fauna., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Owing to its systemic mode-of-action and ease of application, lithium chloride (LiCl) is an ideal varroacide for the control of Varroa destructor infestations in honey bee colonies. To better understand how LiCl functions within a colony, we screened different parts of honey bee anatomy for lithium accumulation. We wanted to elucidate the time-dependent effects of LiCl on V. destructor and its metabolism within honey bees when they were fed continuous LiCl treatments, as well as evaluate potential adverse effects such as accumulation in the hypopharyngeal glands of nurse bees, which could negatively impact queens and larvae., Results: Cage experiments reveal rapid acaricidal onset, with >95% mite mortality within 48 h of treatment. Bee hemolymph analysis supports these observations, showing a rapid increase in lithium concentration within 12 h of treatment, followed by stabilization at a constant level. Lithium accumulates in the rectum of caged bees (≤475.5 mg kg -1 after 7 days of feeding 50 mm LiCl), reflecting the bees' metabolic and excretion process. Despite concerns about potential accumulation in hypopharyngeal glands, low lithium levels of only 0.52 mg kg -1 suggest minimal risk to the queen and 1 st - and 2 nd -instar larvae. Cessation of LiCl treatment results in a rapid decline in mite mortality in the first 5 days, which increases again thereafter, resulting in mite mortality of 77-90% after 10 days., Conclusion: These findings help optimize LiCl application in colonies to achieve high Varroa mortality without unwanted adverse effects and provide important baseline data for future registration. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

The western honey bee (Apis mellifera) is severely impacted by the parasitic Tropilaelaps mercedesae mite, which has the capacity to outcompete Varroa destructor mites (the current leading cause of colony losses) and more rapidly overwhelm colonies. While T. mercedesae is native to Asia, it has recently expanded its geographic range and has the potential to devastate beekeeping worldwide if introduced to new regions. Our research exploited the dependence of T. mercedesae on developing honey bees (brood) by combining a cultural technique (brood break) with U.S. registered chemical products (oxalic acid or formic acid) to manage T. mercedesae infestation. To evaluate this approach, we compared four treatment groups: (1) Brood Break; (2) Brood Break + Formic Acid (FormicPro ® ); (3) Brood Break + Oxalic Acid dribble (Api-Bioxal ® ); and (4) untreated Control. We found that the mite infestation rate of worker brood in Control colonies rose from 0.4 to 15.25% over 60 days, whereas all other treatment groups had infestation rates under 0.11% on Day 60. Mite fall assessments showed similar results, whereby Control colonies had 15.48 mites fall per 24 h on day 60 compared to less than 0.2 mites for any other treatment group. Evaluation of colony strength revealed that Brood Break + Formic Acid colonies had slightly reduced adult honey bee populations. No treatment eliminated all mites, so additional measures may be needed to eradicate T. mercedesae if detected in countries that do not currently have T. mercedesae., (© 2024. The Author(s).)

Varroa destructor is one of the greatest threats to Apis mellifera worldwide and if left untreated will kill a colony in less than three years. A Varroa-resistant population from Gotland, Sweden, has managed to survive for 25 years with little to no Varroa treatment by reducing the mite's reproductive success. The underlying mechanisms of this trait is currently not known, though previous research indicates that it is the honey bee brood, and not adult bee influence, that contributes to this phenotype. As the mite's own reproduction is synchronized with the brood's development though the interception of brood pheromones, it is possible that a change in pheromone profile would disrupt the mite's reproductive timing. To investigate this, we characterized the brood ester pheromone (BEP) profile of our resistant Gotland population compared to a non-resistant control. This was done by extracting and analyzing key cuticular compounds of the BEP using gas chromatography. A significant difference was found immediately after brood capping, indicating a divergence in their pheromonal production at this time point. This is an important step to understanding the mechanisms of the Gotland population's Varroa-resistance and contributes to our global understanding of Varroa destructor infestation and survival., (© 2024. The Author(s).)

Deformed wing virus (DWV), which is an important honey bee virus transmitted by Varroa destructor (V. destructor) , causes colony losses in honey bee colonies. This study aimed to investigate the prevalence and genetic diversity of DWV in honey bees in Türkiye and to determine the role of V. destructor in the transmission of the genetic variants of DWV. Honey bee samples were collected from 62 apiaries, by simple random sampling, during March 2022 and April 2023. The presence of V. destructor in collected bee samples was examined using a stereo microscope. Real-time RT-PCR was used for the detection of DWV-A and DWV-B ( Varroa destructor virus-1 (VDV-1)) viruses. Genetic characterisation of the positive samples was conducted by sequencing polyprotein genomic region. Considering the V. destructor infestation rate of 3% as relevant, out of the 62 apiaries examined, 17 (27.4%) were positive. However, DWV-A and VDV-1 specific RNA was not detected in V. destructor samples. VDV-1 specific RNA was detected in 6.5% (4/62) of the apiaries, whereas DWV-A was not detected in the sampled apiaries. Phylogenetic analysis showed that isolates detected in this study were located in a separate cluster from previously characterised DWV-A and VDV-1 isolates. According to RDP4 and GARD analyses, DWV-VDV-1 recombination breakpoints were detected in field isolates. To the best our knowledge, this is the first report of the presence of VDV-1-DWV recombinants in Türkiye. Further studies are needed to determine the impact of VDV-1-DWV recombinants and their virological and antigenic properties.

Background: The parasitic mite, Varroa destructor has posed a threat to the health and survival of European honey bees, Apis mellifera worldwide. There is a prevailing belief that small comb cells could provide a management tool against Varroa mites. However, the hypothesis that smaller cells can impede Varroa reproduction has not been fully tested. Here, we tested this hypothesis under laboratory conditions by using two distinct Varroa in vitro rearing systems: one involved gelatin capsules of different sizes, specifically size 00 (0.95 mL) versus size 1 (0.48 mL), and the second consisted of brood comb cells drawn on 3D printed foundations with varying cell sizes, ranging from 5.0 mm to 7.0 mm at 0.5 mm intervals., Results: The results showed that mother mites in size 00 cells had significantly lower fecundity and fertility compared to those in size 1 cells. Interestingly, the reproductive suppression in larger cells could be reversed by adding an extra worker larva. Similarly, gonopore size of mother mites was smaller in size 00 cells, but restored with another host larva. Furthermore, both the fecundity and fertility of mother mites decreased linearly with the size of brood comb cells., Conclusions: Our results suggest that the reproduction of V. destructor is hindered by larger cells, possibly because larger brood cells disperse or weaken host volatile chemical cues that are crucial for Varroa reproduction. The insights derived from this study are expected to hold significant implications for the implementation of Varroa management programs. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Most published data on mite infestation rates in semi-arid regions have been collected over only 3 or 4 months during a specific period of the year. Therefore, the aim of this study was to observe parasite-host dynamics of hygienic and non-hygienic Africanized bee colonies considering environmental factors that may influence Varroa destructor mite infestation rates in a semi-arid region. To this end, the brood puncture method was applied to 37 colonies, forming two groups, namely G1, encompassing 16 hygienic colonies, and G2, comprising 21 non-hygienic colonies. After forming the groups, 300 worker bees from each colony were examined monthly for mite infestations and the data were correlated with climatological records. The monthly infestation average was considered low, below 10%, except in November, when it reached 12.19% ± 6.45. No statistically significant difference was observed for inter-group infestation rates (P > 0.05). When mite infestation rates were associated with climatic variables, they were linked to colony losses (32%) due to swarming. No significant correlations between hygienic behaviour and parasite infestation rates were noted. Nonetheless, these results support the idea that there is no need to apply acaricides for V. destructor control in Brazil., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Tropilaelaps spp. are invasive mites that cause severe disease in Apis mellifera colonies. The UK has deployed an elaborate surveillance system that seeks to detect these mites early in any invasion to allow the best opportunity to eradicate any incursion. Effective field and laboratory protocols, capable of reliably detecting low numbers of mites, are key to the success of any intervention. Here we compared the efficacy of established field monitoring using brood removal with an uncapping fork, and brood 'bump' methods with novel methods for Tropilaelaps detection modified from Varroa monitoring schemes. In addition, we monitored the efficacy of the laboratory method for screening for mites in hive debris by floating mites in ethanol. Our results clearly indicated that novel methods such as uncapping infested brood with tweezers, catching mite drop using sticky traps and rolling adult bees in icing sugar were all significantly more likely to detect Tropilaelaps than existing methods such as using an uncapping fork on infested brood, or the brood 'bump' method. Existing laboratory protocols that sieved hive debris and then floated the mite containing layer failed to detect Tropilaelaps mites and new efficacious protocols were developed. Our results demonstrated that the national surveillance protocols for Tropilaelaps mite detection required modification to improve the early detection of this damaging invasive mite., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Gill et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The interaction between environmental factors affecting honey bees is of growing concern due to their potential synergistic effects on bee health. Our study investigated the interactive impact of Varroa destructor and chlorothalonil on workers' survival, fat body morphology, and the expression of gene associated with detoxification, immunity, and nutrition metabolism during their adult stage. We found that both chlorothalonil and V. destructor significantly decreased workers' survival rates, with a synergistic effect observed when bees were exposed to both stressors simultaneously. Morphological analysis of fat body revealed significant alterations in trophocytes, particularly a reduction in vacuoles and granules after Day 12, coinciding with the transition of the bees from nursing to other in-hive work tasks. Gene expression analysis showed significant changes in detoxification, immunity, and nutrition metabolism over time. Detoxification genes, such as CYP9Q2, CYP9Q3, and GST-D1, were downregulated in response to stressor exposure, indicating a potential impairment in detoxification processes. Immune-related genes, including defensin-1, Dorsal-1, and Kayak, exhibited an initially upregulation followed by varied expression patterns, suggesting a complex immune response to stressors. Nutrition metabolism genes, such as hex 70a, AmIlp2, VGMC, AmFABP, and AmPTL, displayed dynamic expression changes, reflecting alterations in nutrient utilization and energy metabolism in response to stressors. Overall, these findings highlight the interactive and dynamic effects of environmental stressor on honey bees, providing insights into the mechanisms underlying honey bee decline. These results emphasize the need to consider the interactions between multiple stressors in honey bee research and to develop management strategies to mitigate their adverse effects on bee populations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Purpose: Nosemosis is a disease that infects both Western honeybees (Apis mellifera L.) and Asian honeybees (Apis cerana) and causes colony losses and low productivity worldwide. In order to control nosemosis, it is important to determine the distribution and prevalence of this disease agent in a particular region. For this purpose, a national study was conducted to assess the prevalence of Nosema ceranae and N. apis throughout Türkiye, to perform network analyses of the parasites, and to determine the presence of nosemosis., Methods: In this study which aimed to assess the prevalence of N. apis and N. ceranae in different colony types and regions where beekeeping is intensive in Türkiye, specimens were collected from hives with no clinical signs., Results: A total of 1194 Western honeybee colonies in 400 apiaries from 40 provinces of Türkiye were examined by microscopic and molecular techniques. Nosemosis was found in all of 40 provinces. The mean prevalence ratio was 64.3 ± 3.0, with 95% CI in apiaries and 40.5 ± 2.9, 95% CI in hives. Nosema ceranae DNA was detected in all of positive hives, while N. ceranae and N. apis co-infection was detected in only four colonies., Conclusion: This study showed that nosemosis has spread to all provinces, and it is common in every region of Türkiye. All of the N. ceranae or N. apis samples examined were 100% identical within themselves. Network analysis showed that they were within largest haplotype reported worldwide., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Gut microbial communities confer protection against natural pathogens in important pollinators from the genera Bombus and Apis. In commercial species B. terrestris and B. impatiens, the microbiota increases their resistance to the common and virulent trypanosomatid parasite Crithidia bombi. However, the mechanisms by which gut microorganisms protect the host are still unknown. Here, we test two hypotheses: microbiota protect the host (1) through stimulation of its immune response or protection of the gut epithelium and (2) by competing for resources with the parasite inside the gut. To test them, we reduced the microbiota of workers and then rescued the microbial community by feeding them with microbiota supplements. We then exposed them to an infectious dose of C. bombi and characterised gene expression and gut microbiota composition. We examined the expression of three antimicrobial peptide genes and Mucin-5AC, a gene with a putative role in gut epithelium protection, using qPCR. Although a protective effect against C. bombi was observed in bumblebees with supplemented microbiota, we did not observe an effect of the microbiota on gene expression that could explain alone the protective effect observed. On the other hand, we found an increased relative abundance of Lactobacillus bacteria within the gut of infected workers and a negative correlation of this genus with Gilliamella and Snodgrassella genera. Therefore, our results point to a displacement of bumblebee endosymbionts by C. bombi that might be caused by competition for space and nutrients between the parasite and the microbiota within the gut., (© 2024 John Wiley & Sons Ltd.)

Microsporidia are obligate intracellular parasites of the Fungal Kingdom that cause widespread infections in nature, with important effects on invertebrates involved in food production systems. The two microsporidian species Vairimorpha (Nosema) ceranae (and the less common Vairimorpha (Nosema) apis) can cause individual disease in honey bees and contribute to colony collapse. The efficacy, safety, and availability of fumagillin, the only drug currently approved to treat microsporidia infection in bees, is uncertain. In this review, we will discuss some of the most promising alternative strategies for the mitigation of Vairimorpha spp. with an emphasis on infection by V. ceranae, now the dominant species infecting bees. We will focus on pharmacologic interventions where the mechanism of action is known and examine both pathogen-directed and host-directed approaches. As limiting toxicity to host cells has been especially emphasized in treating bees that are already facing numerous stressors, strategies that disrupt pathogen-specific targets may be especially advantageous. Therefore, efforts to increase the knowledge and tools for facilitating the discovery of such targets and pharmacologic agents directed against them should be prioritized., (© 2024 International Society of Protistologists.)

Background: The Varroa mite (Varroa destructor) is considered to be the greatest threat to apiculture worldwide. RNA interference (RNAi) using double-stranded RNA (dsRNA) as a gene silencing mechanism has emerged as a next-generation strategy for mite control., Results: We explored the impact of a dsRNA biopesticide, named vadescana, designed to silence the calmodulin gene in Varroa, on mite fitness in mini-hives housed in a laboratory. Two dosages were tested: 2 g/L dsRNA and 8 g/L dsRNA. Vadescana appeared to have no effect on mite survival, however, mite fertility was substantially reduced. The majority of foundress mites exposed to vadescana failed to produce any offspring. No dose-dependent effect of vadescana was observed, as both the low and high doses inhibited mite reproduction equally well in the mini-hives and neither dose impacted pupal survival of the honey bee. Approximately 95% of bee pupae were alive at uncapping across all treatment groups., Conclusion: These findings suggest that vadescana has significant potential as an effective alternative to conventional methods for Varroa control, with broader implications for the utilization of RNAi as a next-generation tool in the management of pest species. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Background: The study was conducted in Pawe district from Benishangul-Gumuz and Jawi and Fagita Lekoma districts from the Amhara region to investigate major honeybee pests, predators and diseases., Methods: Using a purposive sampling technique, 183 households were interviewed, and 240 samples were collected for laboratory analysis of bee disease; data were analysed using descriptive statistics., Results: The share of hive types owned by sampled respondents was 88.6%; overall, 1.1% and 10.3% were traditional, transitional and modern beehives, respectively. About 92% of the sample respondents acquired their base colonies by catching swarm bees on the apex of trees. The majority of beekeepers executed external inspections of their colony, whereas only 50% carried out internal inspections. Based on the responses of beekeepers, around 48.9%, 56.3% and 23.1% of colonies absconded every year from Pawe, Jawi and Fagita Lekoma districts, respectively. Ants, wax moths, bee lice, beetles, spiders, birds, monkeys and honey badgers were the major honeybee pests and predators discovered in study areas in decreasing order. Concerning the incidence of Varroa mites, Nosema apis and amoeba disease, 27.5%, 60% and 71.6% of samples showed positive results in study locations, respectively., Conclusions: From this result, we observed that ants, wax moths, bee lice, beetles, spiders, birds, monkeys and honey badgers were the major honeybee pests and predators. The prevalence of amoeba disease was comparatively higher in highland areas and in the summer season. This finding suggests the need for the alertness of beekeepers in controlling bee disease and pests and strengthening bee colonies through seasonal colony management. There should be a strict quarantine, and check-up undertaken when a new colony is purchased from one region to another is essential., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

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.

Highbush blueberry pollination depends on managed honey bees (Apis mellifera) L. for adequate fruit sets; however, beekeepers have raised concerns about the poor health of colonies after pollinating this crop. Postulated causes include agrochemical exposure, nutritional deficits, and interactions with parasites and pathogens, particularly Melisococcus plutonius [(ex. White) Bailey and Collins, Lactobacillales: Enterococcaceae], the causal agent of European foulbrood disease, but other pathogens could be involved. To broadly investigate common honey bee pathogens in relation to blueberry pollination, we sampled adult honey bees from colonies at time points corresponding to before (t1), during (t2), at the end (t3), and after (t4) highbush blueberry pollination in British Columbia, Canada, across 2 years (2020 and 2021). Nine viruses, as well as M. plutonius, Vairimorpha ceranae, and V. apis [Tokarev et al., Microsporidia: Nosematidae; formerly Nosema ceranae (Fries et al.) and N. apis (Zander)], were detected by PCR and compared among colonies located near and far from blueberry fields. We found a significant interactive effect of time and blueberry proximity on the multivariate pathogen community, mainly due to differences at t4 (corresponding to ~6 wk after the beginning of the pollination period). Post hoc comparisons of pathogens in near and far groups at t4 showed that detections of sacbrood virus (SBV), which was significantly higher in the near group, not M. plutonius, was the primary driver. Further research is needed to determine if the association of SBV with highbush blueberry pollination is contributing to the health decline that beekeepers observe after pollinating this crop., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Honey bees (Apis mellifera L.) are the premier agricultural pollinators with direct ecological value and are key to some agro-economies. Major factors have negatively impacted honey bee health in the past 2 decades with Varroa (Varroa destructor Anderson and Trueman) infestation rising as a principal predictor of colony mortality. A key strategy deployed in Varroa management is breeding for resistant honey bee populations that can maintain comparable levels of productivity as nonresistant populations. In this study, we examine one such population, Hilo honey bees, within the context of a common garden contrast with a commercial population in a stationary honey production operation. We compare colony survival, health, yield, and profit outcomes to show how this specific breeding population retains a profit value in honey production operations while maintaining higher survival and lower Varroa infestation levels than the commercial population. This information can be used by commercial beekeepers to make best management practice decisions and inspire further work examining what trade-offs, if any, are present in this Varroa-resistant population., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

This is the first study providing long-term data on the dynamics of bees and wasps and their parasitoids for the evidence-based management of reed beds. Ten years ago, we identified Lipara (Chloropidae) - induced galls on common reed (Phragmites australis, Poaceae) as a critically important resource for specialized bees and wasps (Hymenoptera: Aculeata). We found that they were surprisingly common in relatively newly formed anthropogenic habitats, which elicited questions about the dynamics of bees and wasps and their parasitoids in newly formed reed beds of anthropogenic origin. Therefore, in the winter and spring of 2022/23, we sampled reed galls from the same set of reed beds of anthropogenic and natural origin as those in 2012/13. At 10 sites, the number of sampled galls was similar in both time periods (80-122% of the value from 2012/13); 12 sites experienced a moderate decline (30-79% of the value from 2012/13), and the number of galls at six sampling sites was only 3-23% of their abundance in 2012/13. Spontaneous development was associated with increasing populations. After 10 years of spontaneous development, the populations of bees and wasps (including their parasitoids) bound to Lipara-induced reed galls increased in abundance and species richness or remained at their previous levels, which was dependent on the sampling site. The only identified threat consisted of reclamation efforts. The effects of habitat age were limited, and the assemblages in habitats of near-natural and anthropogenic origin largely overlapped. However, several species were consistently present at lower abundances in the anthropogenic habitats and vice versa. In conclusion, we provided evidence-based support for the establishment of oligotrophic reed beds of anthropogenic origin as management tools providing sustainable habitats for specialized reed gall-associated aculeate hymenopteran inquilines, including the threatened species., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Varroa mite infestation poses a severe threat to honeybee colonies globally. This study investigates the feasibility of utilizing the HS-Cam and machine learning techniques for Varroa mite counting. The methodology involves image acquisition, dimensionality reduction through Principal Component Analysis (PCA), and machine learning-based segmentation and classification algorithms. Specifically, a k-Nearest Neighbors (kNNs) model distinguishes Varroa mites from other objects in the images, while a Support Vector Machine (SVM) classifier enhances shape detection. The final phase integrates a dedicated counting algorithm, leveraging outputs from the SVM classifier to quantify Varroa mite populations in hyperspectral images. The preliminary results demonstrate segmentation accuracy exceeding 99% and an average precision of 0.9983 and recall of 0.9947 across all the classes. The results obtained from our machine learning-based approach for Varroa mite counting were compared against ground-truth labels obtained through manual counting, demonstrating a high degree of agreement between the automated counting and manual ground truth. Despite working with a limited dataset, the HS-Cam showcases its potential for Varroa counting, delivering superior performance compared to traditional RGB images. Future research directions include validating the proposed hyperspectral imaging methodology with a more extensive and diverse dataset. Additionally, the effectiveness of using a near-infrared (NIR) excitation source for Varroa detection will be explored, along with assessing smartphone integration feasibility.

In June 2022, an exotic pest of the European honeybee (Apis mellifera), the varroa mite (Varroa destructor), was detected in surveillance hives at the Port of Newcastle, New South Wales (NSW). Previously, Australia remained the only continent free of the varroa mite. In September 2023, the National Management Group decided to shift the focus of the response from eradication to management. It is estimated that the establishment of varroa mite in Australia could lead to more than $70 million in losses each year due to greatly reduced pollination services. Currently, there are no reported studies on the epidemiology of varroa mite in NSW because it is such a recent outbreak, and there is little knowledge of the factors associated with the presence of V. destructor in the Australian context. We sourced publicly available varroa mite outbreak reports from June 22 to December 19, 2022, to determine if urbanization, land use, and distance from the incursion site are associated with the detection of varroa mite infestation in European honeybee colonies in NSW. The outcome investigated was epidemic day, relative to the first detected premises (June 22, 2022). The study population was comprised of 107 premises, which were declared varroa-infested. The median epidemic day was day 37 (July 29, 2022), and a bimodal distribution was observed from the epidemic curve, which was reflective of an intermittent source pattern of spread. We found that premises were detected to be infected with varroa mite earlier in urban areas [median epidemic day 25 (July 17, 2022)] compared to rural areas [median epidemic day 37.5 (July 29, 2022)]. Infected premises located in areas without cropping, forests, and irrigation were detected earlier in the outbreak [median epidemic days 23.5 (July 15, 2022), 30 (July 22, 2022), and 15 (July 7, 2022), respectively] compared to areas with cropping, forests, and irrigation [median epidemic days 50 (August 11, 2022), 43 (August 4, 2022), and 47 (August 8, 2022), respectively]. We also found that distance from the incursion site was not significantly correlated with epidemic day. Urbanization and land use are potential factors for the recent spread of varroa mite in European honeybee colonies in NSW. This knowledge is essential to managing the current varroa mite outbreak and preventing future mass varroa mite spread events.

The worldwide dispersal of the ectoparasitic mite Varroa destructor from its Asian origins has fundamentally transformed the relationship of the honey bee (Apis mellifera) with several of its viruses, via changes in transmission and/or host immunosuppression. The extent to which honey bee-virus relationships change after Varroa invasion is poorly understood for most viruses, in part because there are few places in the world with several geographically close but completely isolated honey bee populations that either have, or have not, been exposed long-term to Varroa, allowing for separate ecological, epidemiological, and adaptive relationships to develop between honey bees and their viruses, in relation to the mite's presence or absence. The Azores is one such place, as it contains islands with and without the mite. Here, we combined qPCR with meta-amplicon deep sequencing to uncover the relationship between Varroa presence, and the prevalence, load, diversity, and phylogeographic structure of eight honey bee viruses screened across the archipelago. Four viruses were not detected on any island (ABPV-Acute bee paralysis virus, KBV-Kashmir bee virus, IAPV-Israeli acute bee paralysis virus, BeeMLV-Bee macula-like virus); one (SBV-Sacbrood virus) was detected only on mite-infested islands; one (CBPV-Chronic bee paralysis virus) occurred on some islands, and two (BQCV-Black queen cell virus, LSV-Lake Sinai virus,) were present on every single island. This multi-virus screening builds upon a parallel survey of Deformed wing virus (DWV) strains that uncovered a remarkably heterogeneous viral landscape featuring Varroa-infested islands dominated by DWV-A and -B, Varroa-free islands naïve to DWV, and a refuge of the rare DWV-C dominating the easternmost Varroa-free islands. While all four detected viruses investigated here were affected by Varroa for one or two parameters (usually prevalence and/or the Richness component of ASV diversity), the strongest effect was observed for the multi-strain LSV. Varroa unambiguously led to elevated prevalence, load, and diversity (Richness and Shannon Index) of LSV, with these results largely shaped by LSV-2, a major LSV strain. Unprecedented insights into the mite-virus relationship were further gained from implementing a phylogeographic approach. In addition to enabling the identification of a novel LSV strain that dominated the unique viral landscape of the easternmost islands, this approach, in combination with the recovered diversity patterns, strongly suggests that Varroa is driving the evolutionary change of LSV in the Azores. This study greatly advances the current understanding of the effect of Varroa on the epidemiology and adaptive evolution of these less-studied viruses, whose relationship with Varroa has thus far been poorly defined., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Lopes et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In recent years, managed honey bee colonies have been suffering from an increasing number of biotic and abiotic stressors, resulting in numerous losses of colonies worldwide. A pan-European study, EPILOBEE, estimated the colony loss in Belgium to be 32.4% in 2012 and 14.8% in 2013. In the current study, absolute viral loads of four known honey bee viruses (DWV-A, DWV-B, AmFV, and BMLV) and three novel putative honey bee viruses (Apis orthomyxovirus 1, apthili virus, and apparli virus) were determined in 300 Flemish honey bee samples, and associations with winter survival were determined. This revealed that, in addition to the known influence of DWV-A and DWV-B on colony health, one of the newly described viruses (apthili virus) shows a strong yearly difference and is also associated with winter survival. Furthermore, all scrutinized viruses revealed significant spatial clustering patterns, implying that despite the limited surface area of Flanders, local virus transmission is paramount. The vast majority of samples were positive for at least one of the seven investigated viruses, and up to 20% of samples were positive for at least one of the three novel viruses. One of those three, Apis orthomyxovirus 1, was shown to be a genuine honey bee-infecting virus, able to infect all developmental stages of the honey bee, as well as the Varroa destructor mite. These results shed light on the most prevalent viruses in Belgium and their roles in the winter survival of honey bee colonies., Importance: The western honey bee ( Apis mellifera ) is a highly effective pollinator of flowering plants, including many crops, which gives honey bees an outstanding importance both ecologically and economically. Alarmingly high annual loss rates of managed honey bee colonies are a growing concern for beekeepers and scientists and have prompted a significant research effort toward bee health. Several detrimental factors have been identified, such as varroa mite infestation and disease from various bacterial and viral agents, but annual differences are often not elucidated. In this study, we utilize the viral metagenomic survey of the EPILOBEE project, a European research program for bee health, to elaborate on the most abundant bee viruses of Flanders. We complement the existing metagenomic data with absolute viral loads and their spatial and temporal distributions. Furthermore, we identify Apis orthomyxovirus 1 as a potentially emerging pathogen, as we find evidence for its active replication honey bees., Competing Interests: The authors declare no conflict of interest.

Winter mortality of honey bee colonies represents a major source of economic loss for the beekeeping industry. The objectives of this prospective study were to estimate the incidence risk of winter colony mortality in southwestern Quebec, Canada and to evaluate and quantify the impact of the associated risk factors. A total of 242 colonies from 31 apiaries was selected for sampling in August 2017. The presence of Varroa destructor, Vairimorpha (Nosema) spp., Melissococcus plutonius , deformed wing virus (DWV), and viruses of the acute-Kashmir-Israeli complex (AKI complex) was investigated in each colony. Management practices of the various colonies were obtained from a questionnaire. The incidence risk of colony mortality during the winter of 2017-2018 was estimated to be 26.5% [95% confidence interval (CI): 15.4 to 40.3]. In logistic regression modeling of winter mortality in colonies, an interaction was discovered between V. destructor and DWV; the detection of ≥ 1 V. destructor mites per 100 bees was associated with higher odds of mortality (3.46, 95% CI: 1.35 to 8.90) compared to colonies with < 1 mite per 100 bees, but only in DWV-positive colonies. There were more colony losses in apiaries from beekeepers owning 1 to 5 colonies than in apiaries from beekeepers owning over 100 colonies, which suggests that beekeeper experience and/or type of management are important contributors to winter colony mortality. Assuming a causal relationship, the results of this study suggest that up to 9% of all colony mortalities in the population could have been prevented by reducing the level of V. destructor to < 1 mite per 100 bees in all colonies., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

Since the global invasion of the ectoparasitic mite Varroa destructor (Anderson and Trueman), selection of mite-resistant honey bee (Apis mellifera L.) colonies appears challenging and has to date not broadly reduced colony mortality. The low published estimated heritability values for mite infestation levels could explain the limited genetic progresses obtained so far. We hypothesize that intercolonial horizontal mite transmission could differentially affect the single colonies located in a given apiary and therefore invisibly bias colony infestation phenotypes. This bias may be lower in regions with lower colony density, providing suitable conditions to set up evaluation apiaries. To verify these hypotheses, we monitored mite infestation and reinvasion in experimental colonies, as well as infestation in neighboring colonies belonging to beekeepers in three areas with variable colony densities in the canton of Bern, Switzerland during three consecutive beekeeping seasons. Mite immigration fluctuated between apiaries and years and significantly contributed to colony infestation level. Depending on apiary and year, 17-48% of the mites present in the experimental colonies at the time of the summer oxalic acid final treatment potentially derived from mite immigration that had occurred since mid-spring. Mite immigration was not linked to local colony density or the infestation levels of beekeepers' colonies located within 2 km. Our results do not prove that apiaries for colony evaluation should necessarily be established in areas with low colony density. However, they highlight the high impact of beekeeping management practices on mite colony infestation levels., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Crithidia bombi is a trypanosomatid parasite that infects several species of bumble bees (Bombus spp.), by adhering to their intestinal tract. Crithidia bombi infection impairs learning and reduces survival of workers and the fitness of overwintering queens. Although there is extensive research on the ecology of this host-pathogen system, we understand far less about the mechanisms that mediate internal infection dynamics. Crithidia bombi infects hosts by attaching to the hindgut via the flagellum, and one previous study found that a nectar secondary compound removed the flagellum, preventing attachment. However, approaches that allow more detailed observation of parasite attachment and growth would allow us to better understand factors mediating this host-pathogen relationship. We established techniques for genetic manipulation and visualization of cultured C. bombi. Using constructs established for Crithidia fasciculata, we successfully generated C. bombi cells expressing ectopic fluorescent transgenes using two different selectable markers. To our knowledge, this is the first genetic modification of this species. We also introduced constructs that label the mitochondrion and nucleus of the parasite, showing that subcellular targeting signals can function across parasite species to highlight specific organelles. Finally, we visualized fluorescently tagged parasites in vitro in both their swimming and attached forms, and in vivo in bumble bee (Bombus impatiens) hosts. Expanding our cell and molecular toolkit for C. bombi will help us better understand how factors such as host diet, immune system, and physiology mediate outcomes of infection by these common parasites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Apis mellifera filamentous virus (AmFV) is a double-stranded DNA virus that infects Apis mellifera bees. To our knowledge, this is the first comprehensive study aiming to detect and analyse the genetic diversity and prevalence of AmFV in Korean honeybee colonies. Phylogenetic analysis based on baculovirus repeat open reading frame-N gene (Bro-N) sequences revealed that AmFV isolates from the Republic of Korea (ROK) fell into two distinct lineages, with genetic origins in Switzerland and China, with nucleotide similarities of 98.3% and 98.2%, respectively. Our prevalence analysis demonstrated a noteworthy infection rate of AmFV in 545 honeybee colonies, reaching 33.09% in 2022 and increasing to 44.90% by 2023. Intriguingly, we also detected AmFV in Varroa destructor mites, highlighting their potential role as vectors and carriers of AmFV. The presence of AmFV was correlated with an increased infection rate of sacbrood virus, deformed wing virus, Lake Sinai virus 2, black queen cell virus, and Nosema ceranae in honeybee colonies. These findings provide valuable insight into the prevalence and potential transmission mechanisms of AmFV in honeybee colonies in the ROK. The results of this study may be instrumental in the effective management of viral infections in honeybee apiaries., (© 2024. The Author(s).)

Varroa mites, notorious for parasitizing honeybees, are generally classified as Varroidae. Their extremely modified morphologies and behaviors have led to debates regarding their phylogenetic position and classification as an independent family. In this study, two different datasets were employed to reconstruct the phylogenies of Varroa mites and related Laelapidae species: (1) 9257 bp from the whole 13 mitochondrial protein-coding genes of 24 taxa, (2) 3158 bp from 113 taxa using Sanger sequencing of four nuclear loci. Both mitochondrial and nuclear analyses consistently place Varroa mites within the Laelapidae. Here we propose to place Varroa mites in the subfamily Varroinae stat. nov., which represents a highly morphologically adapted group within the Laelapidae. Ancestral state reconstructions reveal that bee-associated lifestyles evolved independently at least three times within Laelapidae, with most phoretic traits originating from free-living ancestors. Our revised classification and evolutionary analyses will provide new insight into understanding the Varroa mites., (© 2024. The Author(s).)

Varroa mites, scientifically identified as Varroa destructor , pose a significant threat to beekeeping and cause one of the most destructive diseases affecting honey bee populations. These parasites attach to bees, feeding on their fat tissue, weakening their immune systems, reducing their lifespans, and even causing colony collapse. They also feed during the pre-imaginal stages of the honey bee in brood cells. Given the critical role of honey bees in pollination and the global food supply, controlling Varroa mites is imperative. One of the most common methods used to evaluate the level of Varroa mite infestation in a bee colony is to count all the mites that fall onto sticky boards placed at the bottom of a colony. However, this is usually a manual process that takes a considerable amount of time. This work proposes a deep learning approach for locating and counting Varroa mites using images of the sticky boards taken by smartphone cameras. To this end, a new realistic dataset has been built: it includes images containing numerous artifacts and blurred parts, which makes the task challenging. After testing various architectures (mainly based on two-stage detectors with feature pyramid networks), combination of hyperparameters and some image enhancement techniques, we have obtained a system that achieves a mean average precision (mAP) metric of 0.9073 on the validation set.

The grooming behavior of honeybees serves as a crucial auto-protective mechanism against Varroa mite infestations. Compared to Apis mellifera , Apis cerana demonstrates more effective grooming behavior in removing Varroa mites from the bodies of infested bees. However, the underlying mechanisms regulating grooming behavior remain elusive. In this study, we evaluated the efficacy of the auto-grooming behavior between A. cerana and A. mellifera and employed RNA-sequencing technology to identify differentially expressed genes (DEGs) in bee brains with varying degrees of grooming behavior intensity. We observed that A. cerana exhibited a higher frequency of mite removal between day 5 and day 15 compared to A. mellifera , with day-9 bees showing the highest frequency of mite removal in A. cerana . RNA-sequencing results revealed the differential expression of the HTR2A and SLC17A8 genes in A. cerana and the CCKAR and TpnC47D genes in A. mellifera . Subsequent homology analysis identified the HTR2A gene and SLC17A8 gene of A. cerana as homologous to the HTR2A gene and SLC17A7 gene of A. mellifera . These DEGs are annotated in the neuroactive ligand-receptor interaction pathway, the glutamatergic synaptic pathway, and the calcium signaling pathway. Moreover, CCKAR , TpnC47D , HTR2A , and SLC17A7 may be closely related to the auto-grooming behavior of A. mellifera , conferring resistance against Varroa infestation. Our results further explain the relationship between honeybee grooming behavior and brain function at the molecular level and provide a reference basis for further studies of the mechanism of honeybee grooming behavior.

The bee louse Braula spp. had until recently a distribution coincident with its host the honey bee. The adult fly usually attaches to a worker honey bee and steals food from its mouth. However, not all worker bees carry Braula spp. and the mechanism used by Braula spp. to select hosts is not well understood. Using choice remounting bioassays and chemical analyses, we determined host selection and the cues used by B. coeca, a species associated with the African honey bee Apis mellifera scutellata. Braula coeca successfully remounted bees from which they were initially removed and preferred their mandibular gland pheromones (MDG) over those of bees not carrying them. The bee lice did not show any preference for the cuticular hydrocarbons of both types of workers. Chemical analyses of the MDG extracts, revealed quantitative differences between the two categories of workers, with workers carrying B. coeca having more of the queen substance (9-oxo-2(E)-decenoic acid) and worker substance (10-hydroxy-2(E)-decenoic). Braula coeca showed a dose response to the queen substance, indicating its ability to use host derived kairomones as cues that allowed it to benefit from trophallactic dominance by individuals that have a higher probability of being fed by other workers., (© 2024. The Author(s).)

Varroa destructor is one of the most destructive enemies of the honey bee, Apis mellifera all around the world. Several control methods are known to control V. destructor, but the efficacy of several alternative control methods remains unexplored. Irradiation can be one of these unknown solutions but before practical application, the effectiveness, and the physiological effects of ionizing radiation on the host and the parasite are waiting to be tested. Therefore, the objective of our study was to investigate the effects of different doses (15, 50, 100, and 150 Gy) of high-energy X-ray irradiation through mortality rates and hemocyte composition changes in A. mellifera workers and record the mortality rates of the parasite. The mortality rate was recorded during short-term (12, 24, and 48 h) and long-term periods (3, 6, 12, 18, and 24d). The sensitivity of the host and the parasite in case of the higher doses of radiation tested (50, 100, and 150 Gy) been demonstrated by total mortality of the host and 90 % of its parasite has been observed on the 18th day after the irradiation. V. destructor showed higher sensitivity (1.52-times higher than the adult honey bee workers) at the lowest dose (15 Gy). A. mellifera hemocytes were influenced significantly by radiation dosage and the elapsed time after treatment. The higher radiation doses increased plasmatocyte numbers in parallel with the decrease in prohemocyte numbers. On the contrary, the numbers of granulocytes and oencoytes increased in the treated samples, but the putative effects of the different dosages on the recorded number of these hemocyte types could not be statistically proven. In summary, based on the outcome of our study X-ray irradiation can be deemed an effective tool for controlling phoretic V. destructor. However, further research is needed to understand the physiological response of the affected organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

The honeybee ectoparasite Varroa destructor is a major threat to apiculture when evaluating bee diseases and pests. While attempting to control this mite, beekeepers often depend on a small selection of authorized synthetic acaricides, such as flumethrin, which is widely used in Türkiye and globally. However, resistance to flumethrin develops due to incorrect and excessive use. In this study conducted at Ordu Beekeeping Research Institute, trial group were established including an untreated control group and group where flumethrin-based pesticides were applied. Dead varroas collected from pollen traps and live varroas collected from bees were obtained from these trial groups for molecular analysis as positive-negative controls. Varroa samples were collected from provinces representing different regions with intensive beekeeping activities such as Adana, Ankara, Bingöl, Muğla, Ordu, Şanlıurfa, Tekirdağ. Molecular methods were employed to investigate the resistance gene region for pyrethroids (specifically flumethrin) against V. destructor. In our study, individual DNA extractions were performed on dead parasites from colonies subjected to pyrethroid application (resistance negative control) and live parasites (resistance positive control). The DNA samples obtained were used in PCR reactions targeting the region encoding the 925th amino acid of the voltage-gated sodium channel (VGSC) gene, which is responsible for resistance formation. The DNA samples were subjected to gel electrophoresis to observe the amplification products of the expected target region. To examine the nucleotide sequence changes that encode leucine at the 925th amino acid, which is associated with resistance, DNA sequence analysis was applied to the amplification products. Out of 332 V. destructor parasites obtained from different provinces, 279 were analysed using molecular methods. It was observed that 31% of the samples showed sensitivity to flumethrin while 69% exhibited resistance to it. Among the resistant samples: 27% had homozygous isoleucine mutation; 28% had homozygous valine mutation; 2.8% had heterozygous isoleucine mutation; 8.5% had heterozygous valine mutation; and 2.8% had heterozygous methionine mutation, all of which were associated with flumethrin resistance. As a result, the rate of flumethrin resistance in parasites varied between 51% and 94% among different provinces., (© 2024. The Author(s).)

A 2-year field experiment was performed to test lithium chloride, LiCl, application in a normal beekeeping management system. The effect of LiCl on bee larval mortality, beehive weight (honey production) and Varroa mite mortality were tested. Spectrometric quantification of Li on honey and the larval body were made to test the effectiveness of the presence of LiCl. Li was detected in bee larval bodies and in honey over 2 years, from 2018 to 2019. According to the results, no effect of LiCl on mite mortality or bee larval mortality was detected in the first year of application. By assessing the weight variation of beehives, only one LiCl-treated hive showed a significantly higher weight, whereas no other differences were detected between treatments and control. The same trend seen in 2018 was repeated in 2019, while a total bee larval mortality was observed after the first LiCl application, and still no differences in Varroa mite mortality were observed. According to these results, it was concluded that LiCl has no effect on Varroa mite mortality during normal beekeeping practice; furthermore, the recommended amount of treatment (25 mM) had a lethal effect (i.e., total mortality) on larvae following repeated applications., (© 2023. The Author(s).)

In La Réunion, the established honeybee subspecies Apis mellifera unicolor, an endemic subspecies of African lineage, is facing considerable challenges. Since the introduction of the Varroa destructor mite in 2017 high colony losses have been recorded. We investigated the dynamics of V. destructor and two viruses, the Deformed Wing Virus (DWV), known to be transmitted by the mite, and the Chronic Bee Paralysis Virus (CBPV), in A. m. unicolor. Colonies from two apiaries located at 300 and 900 m a.s.l were monitored twice for one year without any acaricide treatment. The brood area, V. destructor infestation rates, DWV and CBPV prevalence and load were recorded monthly. A. m. unicolor maintained brood rearing throughout the year. Varroa destructor infestation resulted in high colony mortality (up to 85 %) and high phoretic mite rates (up to 52 mites per hundred bees). The establishment of DWV in colonies occurred after that of V. destructor and the mite infestation rate had a significant effect on the virus prevalence and load. CBPV appeared only transiently throughout the surveys. The data showed that, in tropical colonies with permanent brood rearing, V. destructor and DWV can reach high levels, but are still subject to seasonal variations that appear to be influenced by environmental conditions. This suggests that beekeeping practices could be adapted by favouring sites and periods for transhumance or acaricide treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Cleptoparasitism, also known as brood parasitism, is a widespread strategy among bee species in which the parasite lays eggs into the nests of the host species. Even though this behavior has significant ecological implications for the dynamics of several species, little is known about the molecular pathways associated with cleptoparasitism. To shed some light on this issue, we used gene expression data to perform a comparative analysis between two solitary neotropical bees: Coelioxoides waltheriae, an obligate parasite, and their specific host Tetrapedia diversipes. We found that ortholog genes involved in signal transduction, sensory perception, learning, and memory formation were differentially expressed between the cleptoparasite and the host. We hypothesize that these genes and their associated molecular pathways are engaged in cleptoparasitism-related processes and, hence, are appealing subjects for further investigation into functional and evolutionary aspects of cleptoparasitism in bees., (© 2024. The Author(s).)

Honey bees (Apis mellifera) are exposed to multiple stressors such as pesticides, lack of forage, and diseases. It is therefore a long-standing aim to develop robust and meaningful indicators of bee vitality to assist beekeepers While established indicators often focus on expected colony winter mortality based on adult bee abundance and honey reserves at the beginning of the winter, it would be useful to have indicators that allow detection of stress effects earlier in the year to allow for adaptive management. We used the established honey bee simulation model BEEHAVE to explore the potential of different indicators such as population size, number of capped brood cells, flight activity, abundance of Varroa mites, honey stores and a brood-bee ratio. We implemented two types of stressors in our simulations: 1) parasite pressure, i.e. sub-optimal Varroa treatment by the beekeeper (hereafter referred as Biotic stress) and 2) temporal forage gaps in spring and autumn (hereafter referred as Environmental stress). Neither stressor type could be detected by bee abundance or honey reserves at the end of the first year. However, all response variables used in this study did reveal early warning signals during the course of the year. The most reliable and useful measures seem to be related to brood and the abundance of Varroa mites at the end of the year. However, while in the model we have full access to time series of variables from stressed and unstressed colonies, knowledge of these variables in the field is challenging. We discuss how our findings can nevertheless be used to develop practical early warning indicators. As a next step in the interactive development of such indicators we suggest empirical studies on the importance of the number of capped brood cells at certain times of the year on bee population vitality., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Groeneveld et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Tropilaelaps mercedesae, an ectoparasitic mite of honeybees, is currently a severe health risk to Apis mellifera colonies in Asia and a potential threat to the global apiculture industry. However, our understanding of the physiological and developmental regulation of this pest remains significantly insufficient. Using ultra-high resolution mass spectrometry, we provide the first comprehensive proteomic profile of T. mercedesae spanning its entire post-embryonic ontogeny, including protonymphs, deutonymphs, mature adults, and reproductive mites. Consequently, a total of 4,422 T. mercedesae proteins were identified, of which 2,189 proteins were significantly differentially expressed (FDR < 0.05) throughout development and maturation. Our proteomic data provide an important resource for understanding the biology of T. mercedesae, and will contribute to further research and effective control of this devastating honeybee pest., (© 2024. The Author(s).)

The survival of the honey bee (Apis mellifera), which has a crucial role in pollination and ecosystem maintenance, is threatened by many pathogens, including parasites, bacteria, fungi and viruses. The ectoparasite Varroa destructor is considered the major cause of the worldwide decline in honey bee colony health. Although several synthetic acaricides are available to control Varroa infestations, resistant mites and side effects on bees have been documented. The development of natural alternatives for mite control is therefore encouraged. The study aims at exploring the effects of cinnamon and oregano essential oils (EOs) and of a mixed fruit cocktail juice on mite infestation levels and bee colony health. A multi-method study including hive inspection, mite count, molecular detection of fungal, bacterial and viral pathogens, analysis of defensin-1, hymenoptaecin and vitellogenin immune gene expression, colony density and honey production data, was conducted in a 20-hive experimental apiary. The colonies were divided into five groups: four treatment groups and one control group. The treatment groups were fed on a sugar syrup supplemented with cinnamon EO, oregano EO, a 1:1 mixture of both EOs, or a juice cocktail. An unsupplemented syrup was, instead, used to feed the control group. While V. destructor affected all the colonies throughout the study, no differences in mite infestation levels, population density and honey yield were observed between treatment and control groups. An overexpression of vitellogenin was instead found in all EO-treated groups, even though a significant difference was only found in the group treated with the 1:1 EO mixture. Viral (DWV, CBPV and BQCV), fungal (Nosema ceranae) and bacterial (Melissococcus plutonius) pathogens from both symptomatic and asymptomatic colonies were detected., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Narciso et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In recent years, there has been growing concern on the potential weakening of honey bees and their increased susceptibility to pathogens due to chronic exposure to xenobiotics. The present work aimed to study the effects on bees undergoing an infection by Nosema ceranae and being exposed to a frequently used in-hive acaricide, amitraz. To achieve this, newly emerged bees were individually infected with N. ceranae spores and/or received a sublethal concentration of amitraz in their diets under laboratory conditions. Mortality, food intake, total volume excrement, body appearance, and parasite development were registered. Bees exposed to both stressors jointly had higher mortality rates compared to bees exposed separately, with no difference in the parasite development. An increase in sugar syrup consumption was observed for all treated bees while infected bees fed with amitraz also showed a diminishment in pollen intake. These results coupled with an increase in the total number of excretion events, alterations in behavior and body surface on individuals that received amitraz could evidence the detrimental action of this molecule. To corroborate these findings under semi-field conditions, worker bees were artificially infected, marked, and released into colonies. Then, they were exposed to a commercial amitraz-based product by contact. The recovered bees showed no differences in the parasite development due to amitraz exposure. This study provides evidence to which extent a honey bee infected with N. ceranae could potentially be weakened by chronic exposure to amitraz treatment., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Honey bees play a major role in crop pollination but have experienced declining health throughout most of the globe. Despite decades of research on key honey bee stressors (e.g., parasitic Varroa destructor mites and viruses), researchers cannot fully explain or predict colony mortality, potentially because it is caused by exposure to multiple interacting stressors in the field. Understanding which honey bee stressors co-occur and have the potential to interact is therefore of profound importance. Here, we used the emerging field of systems theory to characterize the stressor networks found in honey bee colonies after they were placed in fields containing economically valuable crops across Canada. Honey bee stressor networks were often highly complex, with hundreds of potential interactions between stressors. Their placement in crops for the pollination season generally exposed colonies to more complex stressor networks, with an average of 23 stressors and 307 interactions. We discovered that the most influential stressors in a network-those that substantively impacted network architecture-are not currently addressed by beekeepers. Finally, the stressor networks showed substantial divergence among crop systems from different regions, which is consistent with the knowledge that some crops (e.g., highbush blueberry) are traditionally riskier to honey bees than others. Our approach sheds light on the stressor networks that honey bees encounter in the field and underscores the importance of considering interactions among stressors. Clearly, addressing and managing these issues will require solutions that are tailored to specific crops and regions and their associated stressor networks., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Varroa destructor Oud (Acari: Varroidae) is a harmful ectoparasite of Apis mellifera L. honey bees causing widespread colony losses in Europe and North America. To control populations of these mites, beekeepers have an arsenal of different treatments, including both chemical and nonchemical options. However, nonchemical treatments can be labor intensive, and Varroa has gained resistance to some conventional pesticides, and the use of other chemical treatments is restricted temporally (e.g., cannot be applied during periods of honey production). Thus, beekeepers require additional treatment options for controlling mite populations. The compound 1-allyloxy-4-propoxybenzene (3c{3,6}) is a diether previously shown to be a strong feeding deterrent against Lepidopteran larvae and a repellent against mosquitoes and showed promise as a novel acaricide from laboratory and early field trials. Here we test the effect of the compound, applied at 8 g/brood box on wooden release devices, on honey bees and Varroa in field honey bee colonies located in Maryland, USA, and using a thymol-based commercial product as a positive control. 3c{3,6} had minimal effect on honey bee colonies, but more tests are needed to determine whether it affected egg production by queens. Against Varroa3c{3,6} had an estimated efficacy of 78.5%, while the positive control thymol product showed an efficacy of 91.3%. 3c{3,6} is still in the development stage, and the dose or application method needs to be revisited., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

Honey bee parasites remain a critical challenge to management and conservation. Because managed honey bees are maintained in colonies kept in apiaries across landscapes, the study of honey bee parasites allows the investigation of spatial principles in parasite ecology and evolution. We used a controlled field experiment to study the relationship between population growth rate and virulence (colony survival) of the parasite Varroa destructor (Anderson and Trueman). We used a nested design of 10 patches (apiaries) of 14 colonies to examine the spatial scale at which Varroa population growth matters for colony survival. We tracked Varroa population size and colony survival across a full year and found that Varroa populations that grow faster in their host colonies during the spring and summer led to larger Varroa populations across the whole apiary (patch) and higher rates of neighboring colony loss. Crucially, this increased colony loss risk manifested at the patch scale, with mortality risk being related to spatial adjacency to colonies with fast-growing Varroa strains rather than with Varroa growth rate in the colony itself. Thus, within-colony population growth predicts whole-apiary virulence, demonstrating the need to consider multiple scales when investigating parasite growth-virulence relationships., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

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., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

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., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)