Diagnostic study on major honeybee disease, pests and predators in North Western Ethiopia.

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.

Investigation of landscape risk factors for the recent spread of varroa mite (Varroa destructor) in European honeybee (Apis mellifera) colonies in New South Wales, Australia.

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.

Real-time dynamics of aculeate hymenopteran reed gall inquilines in oligotrophic reed beds of anthropogenic and natural origin.

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.

Varroa Mite Counting Based on Hyperspectral Imaging.

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.

Varroa destructor shapes the unique viral landscapes of the honey bee populations of the Azores archipelago.

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.

Do Varroa destructor (Acari: Varroidae) mite flows between Apis mellifera (Hymenoptera: Apidae) colonies bias colony infestation evaluation for resistance selection?

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.

Varroa destructor and deformed wing virus interaction increases incidence of winter mortality in honey bee colonies.

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.

La mortalité hivernale des colonies d'abeilles est une cause importante de pertes économiques en apiculture. Cette étude prospective visait à estimer le risque d'incidence de mortalité hivernale des colonies d'abeilles et les facteurs de risque associés dans le sud-ouest du Québec (Canada). Au total, 242 colonies provenant de 31 ruches ont été sélectionnées en août 2017. La présence de Varroa destructor, de Vairimorpha (Nosema) spp., de Melissococcus plutonius, du virus des ailes déformées (DWV) et des virus du complexe AKI ont été évalués. Les pratiques de régie ont été obtenues selon un questionnaire. Le risque de mortalité des colonies à l'hiver 2017­2018 a été estimé à 26,5 % (95 % CI : 15,4 à 40,3). Dans un modèle de régression logistique, la détection de ≥ 1 mite de V. destructor par 100 abeilles était associée à des cotes plus élevées de mortalité (3,46, 95 % CI : 1,35 à 8,90) comparativement aux colonies avec < 1 mite par 100 abeilles, mais seulement pour les colonies positives au DWV. Les ruchers d'apiculteurs possédant entre 1 et 5 colonies présentaient une mortalité plus élevée comparativement à ceux d'apiculteurs possédant plus de 100 colonies, suggérant une influence de l'expérience ou du type de régie. En assumant une relation causale, les résultats de cette étude suggèrent que jusqu'à 9 % de toutes les mortalités hivernales observées dans la population auraient pu être prévenues en réduisant le niveau d'infestation par V. destructor à moins d'une mite per 100 abeilles dans toutes les colonies.(Traduit par les auteurs).

Known and novel viruses in Belgian honey bees: yearly differences, spatial clustering, and associations with overwintering loss.

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.

Molecular phylogeny reveals Varroa mites are not a separate family but a subfamily of Laelapidae.

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.

First detection and prevalence of Apis mellifera filamentous virus in Apis mellifera and Varroa destructor in the Republic of Korea.

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.

Analysis of Varroa Mite Colony Infestation Level Using New Open Software Based on Deep Learning Techniques.

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.

Comparison of Brain Gene Expression Profiles Associated with Auto-Grooming Behavior between Apis cerana and Apis mellifera Infested by Varroa destructor.

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.

Higher prevalence of sacbrood virus in Apis mellifera (Hymenoptera: Apidae) colonies after pollinating highbush blueberries.

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.

Effects of natural treatments on the varroa mite infestation levels and overall health of honey bee (Apis mellifera) colonies.

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.

Proteomics profiling of the honeybee parasite Tropilaelaps mercedesae across post-embryonic development.

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.

Brood indicators are an early warning signal of honey bee colony loss-a simulation-based study.

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.

Parasite and virus dynamics in the honeybee Apis mellifera unicolor on a tropical island recently invaded by Varroa destructor.

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.

Heightened sensitivity in high-grooming honey bees (Hymenoptera: Apidae).

Honey bees use grooming to defend against the devastating parasite Varroa destructor Anderson and Trueman. We observed the grooming responses of individual bees from colonies previously chosen for high- and low-grooming behavior using a combination of mite mortality and mite damage. Our aim was to gain insight into specific aspects of grooming behavior to compare if high-grooming bees could discriminate between a standardized stimulus (chalk dust) and a stimulus of live Varroa mites and if bees from high-grooming colonies had greater sensitivity across different body regions than bees from low-grooming colonies. We hypothesized that individuals from high-grooming colonies would be more sensitive to both stimuli than bees from low-grooming colonies across different body regions and that bees would have a greater response to Varroa than a standardized irritant (chalk dust). Individuals from high-grooming colonies responded with longer bouts of intense grooming when either stimulus was applied to the head or thorax, compared to sham-stimulated controls, while bees from low-grooming colonies showed no differences between stimulated and sham-stimulated bees. Further, high-grooming bees from colonies with high mite damage exhibited greater grooming to Varroa than high-grooming colonies with only moderate mite damage rates. This study provides new insights into Varroa-specific aspects of grooming, showing that although a standardized stimulus (chalk dust) may be used to assess general grooming ability in individual bee grooming assays, it does not capture the same range of responses as a stimulus of Varroa. Thus, continuing to use Varroa mites in grooming assays should help select colonies with more precise sensitivity to Varroa.

Neonicotinoid exposure increases Varroa destructor (Mesostigmata: Varroidae) mite parasitism severity in honey bee colonies and is not mitigated by increased colony genetic diversity.

Agrochemical exposure is a major contributor to ecological declines worldwide, including the loss of crucial pollinator species. In addition to direct toxicity, field-relevant doses of pesticides can increase species' vulnerabilities to other stressors, including parasites. Experimental field demonstrations of potential interactive effects of pesticides and additional stressors are rare, as are tests of mechanisms via which pollinators tolerate pesticides. Here, we controlled honey bee colony exposure to field-relevant concentrations of 2 neonicotinoid insecticides (clothianidin and thiamethoxam) in pollen and simultaneously manipulated intracolony genetic heterogeneity. We showed that exposure increased rates of Varroa destructor (Anderson and Trueman) parasitism and that while increased genetic heterogeneity overall improved survivability, it did not reduce the negative effect size of neonicotinoid exposure. This study is, to our knowledge, the first experimental field demonstration of how neonicotinoid exposure can increase V. destructor populations in honey bees and also demonstrates that colony genetic diversity cannot mitigate the effects of neonicotinoid pesticides.

The efficacy of 1-allyloxy-4-propoxybenzene (3c{3,6}) against Varroa destructor mites in honey bee colonies from Maryland, USA.

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.