Adult Honey Bees Research Articles

Comparative Study of Amino Acids in the Different Stages of Honey Bee

Apiculture is the practice of keeping bees as well as the manufacturing of honey and beeswax. Nectar is the main source of carbohydrates for honey bees. The amount of nectar needed pet colony depends on how concentrated the sugars are in the nectar. A worker bee needs 11 mg of dry sugar each day, which translates to about 22 µl of 50% sugar syrup per bee per day. In order to make honeybees grow satisfactorily enough feed must be provided, and feed should have all the necessary nutrients for the growth of honeybees. The important nutrients such as amino acids, lipids, carbohydrates, and requirements of vitamins and minerals are necessary for the growth and development of honeybees. During development, several structural, physiological, and biochemical changes occur takes place in an organism from stage to stage. There is a variation in the quality of all biochemical components. In the present study, the free amino acids from the eggs, larvae, pupae, and adult honeybee Apis cerana indica and also honey have been analyzed. This work was carried out by adopting the technique of paper chromatography. The developed chromatograms were dried and the spots were identified by spraying with (O.IN) Ninhydrin in acetone. The Rf values were calculated and compared with the Rf values of the standard amino acids. The presence of amino acids in different life stages of honeybee and honey were analyzed and compared.

Single and combined exposure to 'bee safe' pesticides alter behaviour and offspring production in a ground-nesting solitary bee (Xenoglossa pruinosa).

Mounting evidence supporting the negative impacts of exposure to neonicotinoids on bees has prompted the registration of novel 'bee-friendly' insecticides for agricultural use. Flupyradifurone (FPF) is a butenolide insecticide that shares the same mode of action as neonicotinoids and has been assessed to be 'practically non-toxic to adult honeybees' using current risk assessment procedures. However, these assessments overlook some routes of exposure specific to wild bees, such as contact with residues in soil for ground-nesters. Co-exposure with other pesticides may also lead to detrimental synergistic effects. In a fully crossed experiment, we assessed the possible lethal and sublethal effects of chronic exposure to two pesticides used on Cucurbita crops, the insecticide Sivanto Prime (FPF) and the fungicide Quadris Top (azoxystrobin and difenoconazole), alone or combined, on solitary ground-nesting squash bees (Xenoglossa pruinosa). Squash bees exposed to Quadris Top collected less pollen per flower visit, while Sivanto-exposed bees produced larger offspring. Pesticide co-exposure induced hyperactivity in female squash bees relative to both the control and single pesticide exposure, and reduced the number of emerging offspring per nest compared to individual pesticide treatments. This study demonstrates that 'low-toxicity' pesticides can adversely affect squash bees under field-realistic exposure, alone or in combination.

Monitoring the Season-Prevalence Relationship of Vairimorpha ceranae in Honey Bees (Apis mellifera) over One Year and the Primary Assessment of Probiotic Treatment in Taichung, Taiwan.

Microsporidiosis, which is caused by the pathogen Vairimorpha ceranae, is a prevalent disease in the honey bee (Apis mellifera) and might lead to significant adult honey bee mortality. In this study, we conducted an annual survey of the mature spore load of V. ceranae in the guts of nurse bees and forager bees in the apiary of National Chung Hsing University (NCHU) in Taiwan. The results indicated that, on average, honey bees hosted approximately 2.13 × 106 mature spore counts (MSCs)/bee in their guts throughout the entire year. The highest number of MSCs was 6.28 × 106 MSCs/bee, which occurred in April 2020, and the lowest number of MSCs was 5.08 × 105 MSCs/bee, which occurred in November 2020. Furthermore, the guts of forager bees had significantly higher (>58%) MSCs than those of nurse bees. To evaluate the potential of the probiotic to treat microsporidiosis, the lactic acid bacterium Leuconostoc mesenteroides TBE-8 was applied to honey bee colonies. A significant reduction (>53%) in MSCs following probiotic treatment was observed, indicating the potential of probiotic treatment for managing microsporidiosis. This research provided information on V. ceranae MSCs in the honey bee gut at NCHU in Taiwan and the MSCs' correlation with the annual season. Furthermore, a potential probiotic treatment for microsporidiosis was assessed for future management.

Fall Treatment with Fumagillin Contributes to an Overwinter Shift in Vairimorpha Species Prevalence in Honey Bee Colonies in Western Canada.

(1) Background: Microsporidiosis (nosemosis) is an intestinal disorder of adult honey bees caused by the microsporidian pathogens Vairimorpha apis and Vairimorpha ceranae. In Canada, fumagillin is an approved antibiotic used to treat this disease. However, the recommended dosage is based on efficacy studies for V. apis, the native pathogen in European honey bees. Since the detection of V. ceranae in Apis mellifera, V. ceranae became more prevalent in managed European honey bees and seems to have replaced V. apis due to yet unknown reasons. (2) Methods: This colony study investigated the efficacy of fumagillin administered in the fall to colonies infected with both V. apis and V. ceranae and its effects on the Vairimorpha species' prevalence overwinter. Spore loads in control and fumagillin-treated colonies were analysed by microscopy; Vairimorpha species prevalence was determined molecularly and infection and treatment effects on colony productivity were assessed. (3) Results: Fall fumagillin treatment was associated with a temporary reduction in spore load, but there was no difference in spore loads between treated and control colonies the following spring. Interestingly, fumagillin-treated colonies had a significantly greater prevalence of V. ceranae relative to V. apis the following spring, suggesting fumagillin is less effective in controlling V. ceranae.

Risk assessment of veterinary treatments in bee brood and adult honey bees

Amitraz and tau-fluvalinate are veterinary treatments administered for the control of Varroa destructor in honey bee colonies. These treatments can generate harmful residues for bees. This field study addresses the issue under two scenarios: 1) administration according to the posology and 2) placing contaminated wax containing the treatments. Tau-fluvalinate (Apistan®) residues were detected in larval stages from 25.34 to 84.72 ng g−1 in scenario 1 and from 29.63 to 173.97 ng g−1 in scenario 2. In addition, it was detected in bee bread in scenario 2 from 55.72 to 573.79 ng g−1. In scenario 1, DMPF + DMF (amitraz metabolites) were found from 37.09 to 171.58 ng g−1 (Amicel®) and from 36.74 to 42.64 ng g−1 (Apivar®) in bee bread. In honey, DMPF + DMF ranged from 19.96 to 145.30 ng g−1 (Amicel®) and from 32.20 to 78.43 ng g−1 from (Apivar®). With these treatments, a significant reduction in the population of Varroa mites (mean ± s.d.) was observed between 95.15 ± 2.56% and 98.29 ± 1.97%. There were significant positive correlations between Amicel®, the population of adult bees and the population of brood during evaluation 2 showing a higher effectiveness. According to both RQ (Risk Quotient) and SRA (Standard Risk Approach) values for adult bees and larvae, there is no risk when considering exposure by contact or by ingestion of bee bread or honey. In scenario 2, there is no risk for most of the larval stages, except in two bee hives treated with tau-fluvalinate, where the SRA values were 1.73 and 1.29%, indicating a moderate risk due to acute oral exposure.

Production, characterization and therapeutic efficacy of egg yolk antibodies specific to Nosema ceranae.

Nosema disease, caused by Nosema ceranae, one of the single-celled fungal microsporidian parasites, is one of the most important and common diseases of adult honey bees. Since fumagillin, which has been used for decades in the control of Nosema disease in honey bees (Apis mellifera), poses a toxic threat and its efficacy against N. ceranae is uncertain, there is an urgent need to develop alternative prophylactic and curative strategies for the treatment of this disease. The main aim of this study was to investigate the therapeutic potential of specific egg yolk immunoglobulins (IgY) on Nosema disease. For this purpose, the presence of N. ceranae was determined by microscopic and PCR methods in honey bees collected from Nosema suspicious colonies by conducting a field survey. Layered Ataks chickens, divided into four groups each containing 20 animals, were vaccinated with live and inactivated vaccines prepared from field isolates of N. ceranae. Eggs were collected weekly for 10 weeks following the last vaccination. IgY extraction was performed using the PEG precipitation method from egg yolks collected from each group, and the purity of the antibodies was determined by SDS-PAGE and Western Blot. The presence of N. ceranae-specific IgYs was investigated by Western Blot and indirect ELISA methods. It was determined that specific IgYs showed high therapeutic efficacy on Nosema disease in naturally infected bee colonies. In addition, honey bees collected from infected colonies were brought to the laboratory and placed in cages with 30 bees each, and the effectiveness of IgYs was investigated under controlled conditions. It was detected that specific IgY reduced the Nosema spore load and the number of infected bees significantly in both the field and experimental study groups treated for seven days. It was concluded that chicken IgYs, an innovative and eco-friendly method, had a significant potential for use as an alternative to antifungal drugs.

Memory consolidation in honey bees is enhanced by down-regulation of Down syndrome cell adhesion molecule and changes its alternative splicing.

Down syndrome cell adhesion molecule (Dscam) gene encodes a cell adhesion molecule required for neuronal wiring. A remarkable feature of arthropod Dscam is massive alternative splicing generating thousands of different isoforms from three variable clusters of alternative exons. Dscam expression and diversity arising from alternative splicing have been studied during development, but whether they exert functions in adult brains has not been determined. Here, using honey bees, we find that Dscam expression is critically linked to memory retention as reducing expression by RNAi enhances memory after reward learning in adult worker honey bees. Moreover, alternative splicing of Dscam is altered in all three variable clusters after learning. Since identical Dscam isoforms engage in homophilic interactions, these results suggest a mechanism to alter inclusion of variable exons during memory consolidation to modify neuronal connections for memory retention.

Impact of odorants on perception of sweetness by honey bees.

Organic volatiles produced by fruits can result in overestimation of sweetness by humans, but it is unknown if a comparable phenomenon occurs in other species. Honey bees collect nectar of varying sweetness at different flowering plants. Bees discriminate sugar concentration and generally prefer higher concentrations; they encounter floral volatiles as they collect nectar, suggesting that they, like humans, could be susceptible to sweetness enhancement by odorant. In this study, limonene, linalool, geraniol, and 6-methyl-5-hepten-2-ol were tested for their ability to alter behaviors related to perception of sweetness by honey bees. Honey bees were tested in the laboratory using proboscis extension response-based assays and in the field using feeder-based assays. In the laboratory assays, 6-methyl-5-hepten-2-ol and geraniol, but neither linalool nor limonene, significantly increased responses to low concentrations of sucrose compared with no odorant conditions in 15-day and 25-day-old adult worker honey bees, but not in 35-day-old bees. Limonene reduced responding in 15-day-old bees, but not in the older bees. There was no odorant-based difference in performance in field assays comparing geraniol and limonene with a no odorant control. The interaction of the tested plant volatiles with sucrose concentration revealed in laboratory testing is therefore unlikely to be a major determinant of nectar choice by honey bees foraging under natural conditions. Because geraniol is a component of honey bee Nasonov gland pheromone as well as a floral volatile, its impact on responses in the laboratory may reflect conveyance of different information than the other odorants tested.

Early-Life Sublethal Thiacloprid Exposure to Honey Bee Larvae: Enduring Effects on Adult Bee Cognitive Abilities.

Honey bees have significant ecological and economic value as important pollinators, but they are continuously exposed to various environmental stressors, including insecticides, which can impair their health and cause colony decline. (1) Background: Cognitive abilities are vital for the functional maintenance of honey bees; however, it remains unknown if chronic, low-dose exposure to thiacloprid during the larval stage impairs the cognitive abilities of emerged adult honey bees. (2) Methods: To explore this question, honey bee larvae were fed 0, 0.5, and 1.0 mg/L thiacloprid during their developmental phase. Then, the cognitive (i.e., olfactory learning and memory) abilities of adult honey bees were quantified to assess the delayed impacts of early-stage thiacloprid exposure on adult honey bee cognition. Neural apoptosis and transcriptomic level were also evaluated to explore the neurological mechanisms underlying these effects. (3) Results: Our results revealed that chronic larval exposure to sublethal thiacloprid impaired the learning and memory abilities of adult honey bees by inducing neuronal apoptosis and transcriptomic alterations. (4) Conclusions: We highlighted a previously unknown impairment caused by thiacloprid in honey bees.

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

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

Chronic larval and adult honey bee laboratory testing: Which dietary additive should be considered when a test substance is not solubilized in acetone?

Chronic toxicity tests on adult and larval honey bees (Apis mellifera) can require the use of dietary additives (solvents, emulsifiers, adjuvants and viscosifier agents) when the active ingredient of plant protection products cannot be dissolved or does not remain stable and homogeneous within the test diets. Acetone is the widely used and accepted solvent allowed within the international regulatory guidelines, but it can be ineffective in keeping certain compounds in solution and can cause toxicity to adults and larvae. In this publication, we present an evaluation of alternative additives in adult and larval diets. Six dietary additives including five solvents (ethanol, isopropanol, n-propanol, propylene glycol and triethylene glycol) and a viscosifier agent (xanthan gum) at five concentrations along with a negative control and a solvent control (acetone) were investigated at seven laboratories. The safe levels for bees were determined for each of the additives used in the 10-day chronic adult and 22-day chronic larval tests. In the 10-day chronic adult study, ethanol and isopropanol were found to be safe at concentrations ≤ 5.0 %, while xanthan gum can be reliably used at concentrations ≤ 0.1 %. Greater variability across laboratories was observed for N-propanol, propylene glycol, and triethylene glycol and these agents may cause mortality when added to diets at concentrations above 0.25–0.5 %. The safe levels of additives to larval diet in the 22-day chronic larval test had a greater variability and were generally lower than what were observed for adult diet. Our results do not recommend the inclusion of ethanol or n-propanol into the larval diet, and isopropanol, propylene glycol, and triethylene glycol may cause mortality at concentrations above 0.25–0.5 %. Safe levels for xanthan gum were more variable than what was observed for adults, but it can be used reliably at concentrations ≤ 0.05 %. Our analyses conclude that several additives can be integrated successfully in honey bee laboratory bioassays at levels that cause low mortality to adults and larvae.

Health status of honeybee colonies (Apis mellifera) and disease-associated risk factors in different agroecological zones of Southwest Ethiopia

A cross-sectional study design was conducted in different agroecological zones of southwest Ethiopia from October 2019 to October 2021. The study aimed to determine the prevalence and associated risk factors for honeybee diseases and pests, as well as the impact of these issues on honeybee colonies and their products. To identify potential risk factors for honeybee disease and pests, a multivariate random effects logistic regression analysis was used. Adult honeybee and brood samples from a total of 384 honeybee colonies were collected and tested using standard laboratory diagnostic methods. The highest prevalence (55.8%) of ants was recorded, followed by wax moths (22.5%) and hive beetles (23.3%). In the current study, the main honeybee diseases observed in the study areas were varroosis (36.5%), bee lice (5.2%), nosemosis (39.6%), amoeba (56%), and chalkbrood (4.5%). However, tracheal mites, sachbrood, and American and European foul brood, were not detected. The agroecological zone (OR = 5.2, 95% CI: 1.75–14.85), type of hive (OR = 2.9, 95% CI: 1.17–17.03), management system (OR = 4.3, 95% CI: 1.23–14.70), and the management of the colony (OR = 3.5, 95% CI: 1.31–9.14) were identified as risk factors for varroosis in these areas. The occurrence of nosemosis in colonies was also influenced by the agroecological zone (OR = 12.2, 95% CI: 3.06–48.54) and colony management (OR = 3.4, 95% CI: 1.59–7.23). The agroecological zone (OR = 10.5, 95% CI: 12.76–22.63) and hive type (OR = 3.0, 95% CI: 1.39–6.36) were the primary risk factors for the occurrence of amoeba in honeybee colonies. However, the occurrence of bee lice (OR = 34.7, 95% CI: 3.96–104.93) and chalkbrood (OR = 4.8, 95% CI: 1.44–13.16) in honeybee colonies was only influenced by the agroecological zone in the study areas. This study demonstrated that losses in honey production in the area are significantly attributed to honeybee disease and pests. Therefore, it is essential to increase public awareness of how honeybee diseases and pests affect honey production and to develop and implement appropriate control measures for these diseases and pests. Furthermore, more studies should be conducted to characterize and isolate other causes of honeybee diseases and pests in various locations.

Evaluation of Molecular Epidemiology of IAPV in Several Regions of Iran.

In this study, the Israeli acute paralysis virus (IAPV), a single-stranded RNA virus, was investigated in honey bee colonies, which had a history of mortality, population decline, and parasitic diseases. Samples (adult honey bees) were collected from 328 apiaries from three provinces (Tehran, Alborz, and Mazandaran) of Iran to detect IAPV. After sample preparation, RNA was extracted and cDNA was synthesized to perform the reverse transcription polymerase chain reaction (RT-PCR) method using a PCR primer pair, and a 185 bp fragment was amplified. The results showed that out of 328 samples, 103 (31.4%) samples were positive, which were from Mazandaran (14.33%), Tehran (8.84%), and Alborz (8.23%) provinces. Subsequently, some of the positive samples were sequenced and a phylogenetic tree was drawn. The phylogenetic tree showed that the virus isolates were divided into two distinct groups, including one group that had a high similarity to the European acute bee paralysis virus (ABPV) and one group that had a high similarity to the Kashmir bee virus. In addition, the sequences of the samples in three regions were separated in a node from the strains of ABPV from Eastern Europe. Since the length of the branch between the Iranian sequences and the different strains of ABPV from Eastern Europe was short, it can be assumed that the sequences from Iran have a common ancestor with the mentioned strains of ABPV from Eastern Europe.

Possible functions of ecdysone signaling reiteratively used in the adult honey bee brain

The European honey bee is a model organism for investigating the molecular and neural bases of the brain underlying social behaviors. Mushroom bodies (MBs) are a higher-order center of memory, learning, and sensory integration in insect brains, and honey bee MBs are a model to study adult neuronal plasticity. In the honey bee, MBs comprise three Class I Kenyon cell (KC) subtypes: large-, middle-, and small-type KCs, which are distinguished based on the size and localization of their somata, and gene expression profiles. One of the unique characteristics of honey bee MBs is that genes for ecdysone signaling are expressed in a spatially and temporarily regulated manner in the adult brain, suggesting that they play a role in the functional specialization of each KC subtype and behavioral control. A recent study reported that the transcription factor Mblk-1/E93, which functions downstream of ecdysone signaling during metamorphosis, targets genes involved in synaptic plasticity underlying memory and learning ability in the adult honey bee brain. On the other hand, the ecdysone receptor (EcR), which is expressed in small-type KCs in the MBs, was reported to target genes involved in lipid metabolism in the brain during foraging flight. The target genes for Mblk-1 and EcR in the adult brains differed from those during metamorphosis, implying that the reiterative use of some transcription factors involved in ecdysone signaling, such as EcR and Mblk-1, has contributed to the acquisition of novel MB functions in Aculeata species, including the honey bee.

Supplementation of honey bee production colonies with a native beneficial microbe mixture.

Honey bee colonies form a complex superorganism, with individual and social immune defences that control overall colony health. Sometimes these defences are not enough to overcome infections by parasites and pathogens. For that reason, several studies have been conducted to evaluate different strategies to improve honey bee health. Anovel alternative that is being studied is the use of beneficial microbes. In a previous study, we isolated and characterised bacterial strains from the native gut microbiota of honey bees. Four Apilactobacillus kunkeei strains were mixed and administered in laboratory models to evaluate their potential beneficial effect on larvae and adult bees. This beneficial microbe mixture was safe; it did not affect the expression of immune-related genes, and it was able to decrease the mortality caused by Paenibacillus larvae infection in larvae and reduced the Nosema ceranae spore number in infected adult honey bees. In the present study, we aimed to delve into the impact of the administration of this beneficial microbe mixture on honey bee colonies, under field conditions. The mixture was administered in sugar syrup using lyophilised bacterial cells or fresh cultures, by aspersion or sprayed and feeder, once a week for three consecutive weeks, in autumn or spring 2015, 2017 and 2019. Colony strength parameters were estimated before the administration, and one and three months later. Simultaneously different samples were collected to evaluate the infection levels of parasites and pathogens. The results showed that administering the beneficial microbe mixture decreased or stabilised the infection by N. ceranae or Varroa destructor in some trials but not in others. However, it failed to improve the colony's strength parameters or honey production. Therefore, field studies can be a game-changer when beneficial microbes for honey bees are tested, and meticulous studies should be performed to test their effectiveness.

Mandibular glands secrete 24-methylenecholesterol into honey bee (Apis mellifera) food jelly

Honey bee (Apis mellifera) workers feed their larvae with food jelly that is secreted by specialized glands in their heads – the hypopharyngeal and the mandibular glands. Food jelly contains all the nutrients the larvae need to develop into adult honey bees, including essential dietary sterols. The main sterol in food jelly, 24-methylenecholesterol (24MC), is pollen-derived and delivered in food jelly to the larvae in a complex with two proteins, major royal jelly protein 1 (MRJP1) and apisim. Whereas the proteins are synthesized in the hypopharyngeal glands, the sterol-secreting gland has not been identified. We here identified the mandibular glands as sterol-secreting gland for food jelly production by direct detection of the four main honey bee sterols (24MC, campesterol, β-sitosterol and isofucosterol). Furthermore, 24MC seems to be specifically enriched in the mandibular glands, thereby ensuring that food jelly contains the amounts of 24MC necessary for complex formation with MRJP1 and apisimin.

Impact of deformed wing virus master variants (DWV-A, DWV-B, and DWV-C) in managed honey bee colonies of Türkiye

Aim: This study aimed to determine the deformed wing virus (DWV) master variants in managed honey bee hives in Central Anatolia and the Mediterranean Regions of Türkiye. Also, the relationship of DWV genotypes circulating in the apiaries with clinical signs observed in honey bee hives was investigated. Materials and Methods: For this study, adult honey bees were collected from the same 25 hives in the spring-summer and autumn seasons of 2019 from the provinces of Aksaray, Isparta, Karaman, Konya and Nigde. DWV-specific nucleic acid and DWV genotypes were detected by DWV real-time RT-PCR assay and ABC assay, respectively. Results: Deformed wing virus infection was detected in each sampling season. While many colonies were without any clinical signs, in some of the apiaries where samples were collected, wing deformity, trembling, paralysis, swelling in the abdomen, loss of productivity, and dead bees were observed. The prevalences of DWV-A, DWV-B, and DWV-C in adult honey bees were 62%, 82%, and 24%, respectively. The dominant genotype detected in bee hives was the DWV-B master variant (98%). Also, the virus load of the DWV-A master variant was high in all of the honey bee hives with wintering losses. Conclusion: In this present study, data on the current status of DWV master variants circulating in Turkey and their impacts on honey bee colonies are reported for the first time. Thus, it is thought that DWV, which causes yield losses at varying rates in every season of the year in Turkish bee hives, should be carefully monitored.

Epidemiology, factors influencing prevalence and level of varroosis infestation (Varroa destructor) in honeybee (Apis mellifera) colonies in different agroecologies of Southwest Ethiopia

Little information is available on the epidemiology of varroosis caused by Varroa mite, Varroa destructor infestation in Ethiopia, although it is a devastating honeybee disease that results in significant economic losses in beekeeping. Therefore, between October 2021 and October 2022, a cross-sectional study was carried out in different agroecology zones in Southwest Ethiopia to determine the prevalence and associated risk factors for varroosis, as well as the effects of this disease on honeybee colonies and honey production. A multivariate logistic regression analysis was performed to identify possible risk factors for the prevalence of V. destructor. A total of 384 adult honeybee and worker or drone brood samples were collected from honeybee colonies and examined using standard diagnostic techniques in the laboratory. The result shows that the prevalence of V. destructor was found to be 39.3% (95% CI 34.44–44.21) and 43.2% (38.27–48.18) in adult honeybees and brood, respectively. The major risk factors for the prevalence of V. destructor in the study areas included agroecology (OR = 5.2, 95% CI 1.75–14.85), type of hive (OR = 2.9, 95% CI 1.17–17.03), management system (OR = 4.3, 95% CI 1.23–14.70), and colony management (OR = 3.5, 95% CI 1.31–9.14). The lower level of colony infestation in adult bees and brood was measured as 1.97 ± 0.14 and 3.19 ± 0.25, respectively. Season, colony status, colony management, and agroecology were among the determinant factors of the level of varroa mite infestation in adult bees and brood. The results of the study demonstrated that honey production losses are largely attributable to V. destructor infestation. Therefore, it is critical to inform the community about the effects of V. destructor on honey production and develop and implement effective management strategies for this disease. In addition, further research should be done to identify and isolate additional factors that contribute to varroosis in honeybees in different regions.

Toxicity of spray adjuvants and tank mix combinations used in almond orchards to adult honey bees (Apis mellifera).

Commercial beekeepers transporting honey bees across the United States to provide almond pollination services have reported honey bee deaths, possibly due to pesticide applications made during crop bloom. Pesticides are often applied as "tank mixes", or mixtures of fungicides and insecticides combined into a single application. Spray adjuvants are often added to tank mixes to improve the application characteristics of a pesticide and include spreaders, stickers, or surfactants. The goal of this research was to determine toxicity of adjuvants to adult worker honey bees, both when applied alone and in adjuvant-pesticide tank mixtures. Field-relevant combinations of formulated products were applied to 3-day-old adult worker honey bees using a Potter Spray Tower, and mortality was assessed 48 h following exposure. Adjuvants tested included Activator-90, Attach, Choice Weather Master, Cohere, Dyne-Amic, Induce, Kinetic, LI 700, Liberate, Nu-Film P, PHT Latron B-1956, and Surf-90; fungicides tested include Luna Sensation (Fluopyram and Trifloxystrobin), Pristine (Pyraclostrobin and Boscalid), Tilt (Propiconazole), and Vangard (Cyprodinil), and insecticides tested include Altacor (Chlorantraniliprole), Intrepid 2F (Methoxyfenozide), and a positive control Mustang Maxx (Zeta-cypermethrin). Results demonstrated that exposure to some adjuvants causes acute honey bee mortality at near-field application rates, both when applied alone and in combination with pesticides. Some adjuvant-pesticide combinations demonstrated increased toxicity compared with the adjuvant alone, while others demonstrated decreased toxicity. A better understanding of adjuvant and adjuvant-pesticide tank mixture toxicity to honey bees will play a key role in informing "Best Management Practices" for pesticide applicators using spray adjuvants during bloom when honey bee exposure is likely.

Acute and sublethal effects of acetamiprid alone and in mixture with emamectin benzoate on honeybee, Apis mellifera

The neonicotinoid acetamiprid (ACE) and emamectin benzoate (EMB) are widely utilized agrochemicals worldwide in crop protection. Most previous studies have evaluated on the toxicity of individual insecticides on honeybees. This study evaluated formulation of combined toxicities of acetamiprid (8%) and emamectin benzoate (2.15%) through oral and contact exposures to Apis mellifera. The binary mixture had a lethal dose (LD50) of 1 × 10−3 µg/bee (oral) and 4 × 10−5 µg/bee (contact) to adult honeybees and a lethal concentration (LC50) of 2 × 10−1 µg/ml to larvae. The mixture's oral and contact hazard quotient were significantly greater than ACE, with values of 17,400 and 5000, respectively. The estimated additive indices (AI) showed that ACE and EMB interacted additively with AI values of 0.047 (oral), 0.01 (contact), and 0.02 (larval). The mixture also reduced the survival of larvae, affected proboscis extension response (PER) scores, and impacted the ability of honeybees to discriminate odors. The authors recommend a field study to investigate the mixture's adverse effects on honeybees further.