Adult Honey Bees Research Articles

Managing the parasitic honey bee mite Tropilaelaps mercedesae through combined cultural and chemical control methods

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.

Megaselia scalaris and Senotainia tricuspis Infesting Apis mellifera: Detection by Quantitative PCR, Genotyping, and Involvement in the Transmission of Microbial Pathogens.

The Megaselia scalaris and Senotainia tricuspis parasitoid flies of the honeybee Apis mellifera were found to infest apiaries of different European and Mediterranean countries but their prevalence and impact on apiary health are little known. Therefore, in this study, quantitative PCR (qPCR)-based methods were developed for their rapid detection directly in hive matrices. The newly developed qPCR assays were targeted at the mitochondrial cytochrome oxidase subunit I (COI) gene for the M. scalaris and the cytochrome B (cytB) gene for the S. tricuspis. The tests were preliminarily applied to 64 samples of adult honeybees and hive debris collected in the Abruzzo and Molise regions, Central Italy, and the Republic of Kosovo showing that both flies occur in the two countries and more frequently in Italy. The positive apiaries in Italy were re-sampled by capturing viable forager bees and isolating emerging flies to carry out the genotyping and analyses aimed at defining if these flies can transmit honeybee pathogens. Genotyping based on the COI and cytB gene sequencing for M. scalaris and S. tricuspis, respectively, identified one S. tricuspis genotype and diverse genotypes of M. scalaris highly similar to those from distant countries. Some fly isolates harbored the DNA or RNA of honeybee microbial pathogens Paenibacillus larvae, deformed wing viruses A and B (DWVA and B), black queen cell virus (BQCV), chronic paralysis virus (CBPV), and Nosema ceranae. The results indicated that these parasites should be efficiently controlled in apiaries by using rapid detection methods to facilitate the large screening studies and early detection.

Differential impact of Paenibacillus infection on the microbiota of Varroa destructor and Apis mellifera

The Western honey bee (Apis mellifera) is a vital agricultural pollinator whose populations are threatened by the parasitic mite Varroa destructor and associated pathogens. While the impact of Paenibacillus species on honey bees, particularly Paenibacillus larvae causing American foulbrood, is documented, their effect on the microbiota of Varroa mites remains unclear. This study aimed to investigate the influence of Paenibacillus sp. on the bacterial communities of Varroa mites and adult honey bees. We hypothesized that Paenibacillus sp. would significantly alter the microbiota of Varroa mites but have minimal effect on that of adult honey bees. Utilizing 16S rRNA sequencing data from a previous study, we reanalyzed samples categorized into four groups based on Paenibacillus sp. infection load: highly infected and lowly infected honey bees (A. mellifera) and mites (V. destructor). Infection status was determined by Paenibacillus sp. read counts, with more than three reads indicating high infection. Microbial diversity was assessed using alpha and beta diversity metrics. Co-occurrence networks were constructed to visualize bacterial community assemblies, and network robustness was evaluated through node addition and removal tests. Keystone taxa were identified based on eigenvector centrality and relative abundance. Highly infected Varroa mites exhibited a significant reduction in alpha diversity and a markedly different bacterial community composition compared to lowly infected mites (p < 0.05). Their bacterial co-occurrence networks showed decreased connectivity and robustness, indicating a disruptive effect of Paenibacillus sp. In contrast, adult honey bees displayed no significant differences in alpha diversity or network structure between highly and lowly infected groups (p > 0.05), suggesting a resilient microbiota. Keystone taxa analysis revealed fewer central species in highly infected Varroa mites, potentially impacting network stability. High Paenibacillus sp. infection is associated with significant alterations in the microbiota of Varroa mites, disrupting bacterial communities and potentially affecting mite physiology. The microbiota of adult honey bees appears more robust against Paenibacillus sp. influence. These findings enhance our understanding of the complex interactions within the "honey bee–mite–microorganism" system and may inform future strategies for managing Varroa mite infestations and associated pathogens.

Metal concentration in honeybees along an urbanization gradient in Central Mexico

Urbanization is rapidly increasing worldwide, leading to rising levels of pollution, one of the major drivers of environmental change; yet little is known about the relationship between urbanization intensity and pollution levels in pollinator taxa. Toxic metals are among the most common contaminants in urban environments, but few data exist on their presence in the flora and fauna of cities in Latin America, one of the world's most urbanized and biologically diverse regions. In this study, we used an urban-rural gradient approach to analyze the relationship between the concentrations of eleven metals present in adult honeybees (Apis mellifera) and the degree of urbanization within twelve landscapes in the metropolitan area of Pachuca, Hidalgo, which forms part of the megalopolis of Mexico City. Metal concentrations were compared with previously reported values contrasting honeybees from urban and rural areas after standardizing urbanization levels among published reports. The concentrations of Ag, Cr, Cu, and Zn in honeybees increased significantly with the degree of urbanization. Urbanization was not found to influence the levels of Al, Ba, Cd, Mn, and Sr in honeybees. The maximum concentrations of six metals in our urban sites (Al, Ba, Cd, Cu, Mn, and Sr) were higher than the maximum values reported for bees in other urban areas. The concentrations of two metals measured in our study (Cr and Zn) were within the range of values previously published for urban areas. Compared to other studies, we did not detect Pb in the body of honeybees. We conclude that the concentrations of Ag, Cr, Cu, and Zn present in honeybees are a quantitative reflection of the degree of urbanization in central Mexico. Our results highlight the need to monitor metal emission sources in this and other areas and investigate their effects on bees and other pollinator taxa.

The selection traits of mite non-reproduction (MNR) and Varroa sensitive hygiene (VSH) show high variance in subsequent generations and require intensive time investment to evaluate

The honey bee ectoparasite Varroa destructor is the main cause of honey bee colony losses worldwide. Over the last decades, several projects have focused on improving the robustness of Apis mellifera against this parasitic mite. Selection traits, such as mite non-reproduction (MNR) and Varroa sensitive hygiene (VSH), are favored selection factors in Varroa resistance projects. VSH is a trait where adult honey bees remove the Varroa-infested brood. During this process, the female mites are arrested in their reproductive cycle leading to a reduction of the Varroa population within the bee colony. From 2019 to 2022, 1402 queens were instrumentally inseminated with single or multiple drones in a breeding program. Colonies headed by these queens were established annually, and the MNR and VSH levels were analyzed. VSH was evaluated in response to cells artificially infested with Varroa, and colonies with high VSH values were used to generate our selected VSH stock. Despite crossing high VSH drones and queens, we measured a remarkable heterogeneity of MNR and VSH in the next generation(s), most likely due to the well-described, high recombination rate in the honey bee genome. When assessed multiple times in the same colony, great variance between measurements was observed. Detailed evaluations of daughter colonies are thus required if selection programs want to breed colonies with reliable VSH traits. This constant need to evaluate all offspring to ensure the desirable resistance traits are present results in high workloads and great expenses in selection programs. Furthermore, such large-scale breeding programs are inefficient due to high fluctuations between measurements and generations, indicating we need to develop new approaches and improved methods for assessing Varroa resistance.

Flumethrin exposure perturbs gut microbiota structure and intestinal metabolism in honeybees (Apis mellifera)

Flumethrin mitigates Varroa’s harm to honeybee colonies; however, its residues in colonies threaten the fitness of honeybee hosts and gut microbiota. Our previous research has shown that flumethrin induces significant physiological effects on honeybee larvae; but the effects of flumethrin on the gut microbiota and metabolism of adult honeybees are still unknown. In this study, 1-day-old honeybees were exposed to 0, 0.01, 0.1, and 1 mg/L flumethrin for 14 days and the impacts of flumethrin on the intestinal system were evaluated. The results showed that exposure to 1 mg/L flumethrin significantly reduced honeybee survival and the activities of antioxidative enzymes (superoxide dismutase and catalase) and detoxification enzymes (glutathione S-transferase) in honeybee heads. Moreover, exposure to 0.01, 0.1, and 1 mg/L flumethrin significantly decreased the diversity of the honeybee gut microbiota. Results from untargeted metabolomics showed that long-term exposure to 0.01, 0.1, and 1 mg/L flumethrin caused changes in the metabolic pathways of honeybee gut microbes. Furthermore, increased metabolism of phenylalanine, tyrosine, and tryptophan derivatives was observed in honeybee gut microbes. These findings underscore the importance of careful consideration in using pesticides in apiculture and provide a basis for safeguarding honeybees from pollutants, considering the effects on gut microbes.

Development of germline progenitors in larval queen honeybee ovaries.

Honeybees (Apis mellifera) are a keystone species for managed pollination and the production of hive products. Eusociality in honeybees leads to much of the reproduction in a hive driven by the queen. Queen bees have two large active ovaries that can produce large numbers of eggs if conditions are appropriate. These ovaries are also active throughout the long lives of these insects, up to 5 years in some cases. Recent studies have indicated that the germline precursors of the adult honeybee queen ovary are organized into 8-cell clusters, joined together by a polyfusome; a cytoplasmic bridge. To understand the origin of these clusters, and trace the development of the honeybee queen ovary, we examined the cell types and regionalization of the developing larval and pupal queen ovaries. We used established (nanos and castor), and novel (odd skipped) gene expression markers to determine regions of the developing ovary. Primordial germline cells develop in the honeybee embryo and are organized into ovary structures before the embryo hatches. The ovary is regionalized by larval stage 3 into terminal filaments and germaria. At this stage, clusters of germline cells in the germaria are joined by fusomes and are dividing synchronously. The origin of the 8-cell clusters in the adult germarium is therefore during larval stages. On emergence, the queen ovary has terminal filaments and germaria but has not yet developed any vitellaria, which are produced after the queen embarks on a nuptial flight. The lack of germaria, and the storing of germline progenitors as clusters, may be adaptions for queen bees to endure the metabolic demands of a nuptial flight, as well as rapidly lay large numbers of eggs to establish a hive.

Full-Length Transcriptome Construction and Systematic Characterization of Virulence Factor-Associated Isoforms in Vairimorpha (Nosema) Ceranae.

Vairimorpha (Nosema) ceranae is a single-cellular fungus that obligately infects the midgut epithelial cells of adult honeybees, causing bee microsporidiosis and jeopardizing bee health and production. This work aims to construct the full-length transcriptome of V. ceranae and conduct a relevant investigation using PacBio single-molecule real-time (SMRT) sequencing technology. Following PacBio SMRT sequencing, 41,950 circular consensus (CCS) were generated, and 25,068 full-length non-chimeric (FLNC) reads were then detected. After polishing, 4387 high-quality, full-length transcripts were gained. There are 778, 2083, 1202, 1559, 1457, 1232, 1702, and 3896 full-length transcripts that could be annotated to COG, GO, KEGG, KOG, Pfam, Swiss-Prot, eggNOG, and Nr databases, respectively. Additionally, 11 alternative splicing (AS) events occurred in 6 genes were identified, including 1 alternative 5' splice-site and 10 intron retention. The structures of 225 annotated genes in the V. ceranae reference genome were optimized, of which 29 genes were extended at both 5' UTR and 3' UTR, while 90 and 106 genes were, respectively, extended at the 5' UTR as well as 3' UTR. Furthermore, a total of 29 high-confidence lncRNAs were obtained, including 12 sense-lncRNAs, 10 lincRNAs, and 7 antisense-lncRNAs. Taken together, the high-quality, full-length transcriptome of V. ceranae was constructed and annotated, the structures of annotated genes in the V. ceranae reference genome were improved, and abundant new genes, transcripts, and lncRNAs were discovered. Findings from this current work offer a valuable resource and a crucial foundation for molecular and omics research on V. ceranae.

Minibrain plays a role in the adult brain development of honeybee (Apis mellifera) workers.

The brain of adult honeybee (Apis mellifera) workers is larger than that of queens, facilitating behavioural differentiation between the castes. This brain diphenism develops during the pharate-adult stage and is driven by a caste-specific gene expression cascade in response to unique hormonal milieus. Previous molecular screening identified minibrain (mnb; DYRK1A) as a potential regulator in this process. Here, we used RNAi approach to reduce mnb transcript levels and test its role on brain diphenism development in honeybees. White-eyed unpigmented cuticle worker pupae were injected with dsRNA for mnb (Mnb-i) or gfp, and their phenotypes were assessed two and 8 days later using classic histological and transcriptomic analyses. After 2 days of the injections, Mnb-i bees showed 98% of downregulation of mnb transcripts. After 8 days, the brain of Mnb-i bees showed reduction in total volume and in the volume of the mushroom bodies (MB), antennal, and optic lobes. Additionally, signs of apoptosis were observed in the Kenyon cells region of the MB, and the cohesion of the brain tissues was affected. Our transcriptomic analyses revealed that 226 genes were affected by the knockdown of mnb transcripts, most of which allowing axonal fasciculation. These results suggest the evolutionary conserved mnb gene has been co-opted for promoting hormone-mediated developmental brain morphological plasticity generating caste diphenism in honeybees.

Early exposure to glyphosate during larval development induces late behavioural effects on adult honey bees

As the most abundant pollinator insect in crops, Apis mellifera is a sentinel species of the pollinator communities. In these ecosystems, honey bees of different ages and developmental stages are exposed to diverse agrochemicals. However, most toxicological studies analyse the immediate effects during exposure. Late effects during adulthood after early exposure to pollutants during larval development are poorly studied in bees. The herbicide glyphosate (GLY) is the most applied pesticide worldwide. GLY has been detected in honey and beebread from hives near treated crops. Alterations in growth, morphogenesis or organogenesis during pre-imaginal development could induce late adverse effects after the emergence. Previous studies have demonstrated that GLY alters honey bee development, immediately affecting survival, growth and metabolism, followed by late teratogenic effects. The present study aims to determine the late impact on the behaviour and physiology of adult bees after pre-imaginal exposure to GLY. For that, we reared brood in vitro or in the hive with sub-chronic exposure to the herbicide with the average detected concentration in hives. Then, all newly emerged bees were reared in an incubator until maturity and tested when they became nurse-aged bees. Three behavioural responses were assessed as markers of cognitive and physiological impairment. Our results show i) decreased sensitivity to sucrose regardless of the rearing procedure, ii) increased choice latency and locomotor alterations during chemotaxis and iii) impaired associative learning. These late toxicity signs could indicate adverse effects on task performance and colony efficiency.

Comparing the efficacy of alcohol wash and powdered sugar methods to dislodge the Varroa mite from the body of the adult honey bee

Comparing the efficacy of alcohol wash and powdered sugar methods to dislodge the Varroa mite from the body of the adult honey bee

Early-Life Sublethal Exposure to Thiacloprid Alters Adult Honeybee Gut Microbiota.

Thiacloprid, a neonicotinoid pesticide, is known to affect the gut microbiome of honeybees, yet studies often focus on immediate alternations during exposure, overlooking long-term microbiological impacts post-exposure. This study investigates the influences of sublethal thiacloprid administered during the larval developmental stage of honeybees on physiological changes and gut microbiota of adult honeybees. We found that thiacloprid exposure increased mortality and sugar intake in emerged honeybees. Using 16S rDNA sequencing, we analyzed intestinal microbial diversity of honeybees at one and six days post-emergence. Our findings reveal a significant but transient disruption in gut microbiota on day 1, with recovery from dysbiosis by day 6. This study emphasizes the importance of evaluating chronic sublethal exposure risks of thiacloprid to protect honeybee health.

A field realistic model to assess the effects of pesticides residues and adulterants on honey bee gene expression.

While studies on the sublethal effects of chemical residues in beeswax on adult honey bees are increasing, the study protocols assessing the impacts on honey bee brood in realistic conditions still need to be investigated. Moreover, little is known about the residue's effect on gene expression in honey bee brood. This study reports the effects of chlorpyriphos-ethyl, acrinathrin and stearin worker pupae exposure through contaminated or adulterated beeswax on the gene expression of some key health indicators, using a novel in vivo realistic model. Larvae were reared in acrinathrin (12.5, 25, 10 and 100 ppb) and chlorpyriphos-ethyl (5, 10, 500 and 5000 ppb) contaminated or stearin adulterated beeswax (3, 4, 5, 6 and 9%) in newly formed colonies to reduce the influence of external factors. On day 11, mortality rates were assessed. Honey bee pupae were extracted from the comb after 19 days of rearing and were analysed for the gene expression profile of four genes involved in the immune response to pathogens and environmental stress factors (Imd, dorsal, domeless and defensin), and two genes involved in detoxifications mechanisms (CYP6AS14 and CYP9Q3). We found no linear relation between the increase in the pesticide concentrations and the brood mortality rates, unlike stearin where an increase in stearin percentage led to an exponential increase in brood mortality. The immune system of pupae raised in acrinathrin contaminated wax was triggered and the expression of CYP6AS14 was significantly upregulated (exposure to 12.5 and 25 ppb). Almost all expression levels of the tested immune and detoxification genes were down-regulated when pupae were exposed to chlorpyrifos-contaminated wax. The exposure to stearin triggered the immune system and detoxification system of the pupae. The identification of substance-specific response factors might ultimately serve to identify molecules that are safer for bees and the ecosystem's health.

Endochitinase from the Microsporidia Nosema ceranae facilitates infection in the honey bee Apis mellifera

Bees are crucial pollinators in terrestrial ecosystems, responsible for 80% of insect-driven pollination and playing a vital role in the pollination of 75% of crops. The honey bee, Apis mellifera, is not only used in honey production but also serves as a pollinator in agriculture. However, A. mellifera faces various challenges, including exposure to pathogens such as the Microsporidia Nosema ceranae, which has been linked to decreased crop yields and colony losses. Nosema ceranae spores infect adult honey bees by penetrating the midgut lumen and invading the cytoplasm of epithelial cells, completing their life cycle. However, the midgut possesses a protective mechanical barrier called the peritrophic matrix, composed of chitin and proteins, which prevents epithelial infection. Nevertheless, N. ceranae overcomes this primary defense mechanism, though the specific mechanisms it employs to cross the peritrophic matrix and reach the midgut epithelium are not yet well understood. This study aimed to investigate the potential role of the predicted endochitinase from N. ceranae to infect bees. We tested the hypothesis that inhibiting the expression of N. ceranae endochitinase through RNA interference would impact the pathogen infection of A. mellifera. Bees treated with dsRNA targeting endochitinase, administered 12 and 24 h after spore inoculation, exhibited suppressed endochitinase gene expression and a decrease in the number of total and viable N. ceranae spores in the midgut. These results indicate that inhibiting the expression of the target gene through RNA interference affects Microsporidia infection, underscoring the importance of this enzyme in the infection process.

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.

Antifungal Activity and Mechanism of Camphor Derivatives against Rhizoctonia solani: A Promising Alternative Antifungal Agent for Rice Sheath Blight.

Rice sheath blight, caused by the fungus Rhizoctonia solani, poses a significant threat to rice cultivation globally. This study aimed to investigate the potential mechanisms of action of camphor derivatives against R. solani. Compound 4o exhibited superior fungicidal activities in vitro (EC50 = 6.16 mg/L), and in vivo curative effects (77.5%) at 500 mg/L were significantly (P < 0.01) higher than the positive control validamycin·bacillus (66.1%). Additionally, compound 4o exhibited low cytotoxicity and acute oral toxicity for adult worker honeybees of Apis mellifera L. Mechanistically, compound 4o disrupted mycelial morphology and microstructure, increased cell membrane permeability, and inhibited both PDH and SDH enzyme activities. Molecular docking and molecular dynamics analyses indicated a tight interaction of compound 4o with PDH and SDH active sites. In summary, compound 4o exhibited substantial antifungal efficacy against R. solani, serving as a promising lead compound for further optimization of antifungal agents.

Effects of tefluthrin and guadipyr on the midgut bacteria of adult Apis mellifera

The objective of this study is to assess the potential impact of tefluthrin and guadipyr on the gut microbial composition and metabolism in adult Apis mellifera ligustica, thereby elucidating the underlying mechanisms of insecticide action and its practical implications for bee protection. In this investigation, A. mellifera were subjected to one of three dietary conditions: (1) control sugar water, (2) tefluthrin-infused sugar water, or (3) guadipyr-infused sugar water. After a 10-day exposure period, genomic DNA from the gut bacteria was extracted. High-throughput sequencing was employed to evaluate the potential influence of tefluthrin and guadipyr treatments on the diversity and abundance of gut bacteria. Among the A. mellifera specimens, a total of twenty species of gut bacteria were identified, spanning across five phyla, six classes, eleven orders, eleven families, and fifteen genera. The dominant phyla within the gut bacterial community were Proteobacteria and Bacteroidetes. In comparison to the control group, both the tefluthrin-treated and deltamethrin-treated groups exhibited alterations in the composition of their gut bacterial flora. At the phylum level, there was a significant decrease in the relative abundance of Cyanobacteria (P < 0.05). On the genus level, the tefluthrin group displayed a significant increase in the relative abundance of Bartonella and Serratia (P < 0.05). In the guadipyr-treated group, the relative abundance of Gilliamella and Frischella increased significantly (P < 0.05), while the relative abundance of norank_o_Chloroplast and Enterobacter decreased significantly (P < 0.05). Further analysis of cluster of orthologous genes predicted functional changes in gut microbial metabolism following tefluthrin exposure but no significant changes after guadipyr exposure. Consequently, exposure to tefluthrin and guadipyr can induce shifts in both the composition and metabolic activity of the gut bacteria in A. mellifera. Notably, the impact of tefluthrin on the gut bacteria of A. mellifera appears to be more pronounced compared to that of guadipyr.

Cognitive limits of larval Drosophila: testing for conditioned inhibition, sensory preconditioning, and second-order conditioning.

Drosophila larvae are an established model system for studying the mechanisms of innate and simple forms of learned behavior. They have about 10 times fewer neurons than adult flies, and it was the low total number of their neurons that allowed for an electron microscopic reconstruction of their brain at synaptic resolution. Regarding the mushroom body, a central brain structure for many forms of associative learning in insects, it turned out that more than half of the classes of synaptic connection had previously escaped attention. Understanding the function of these circuit motifs, subsequently confirmed in adult flies, is an important current research topic. In this context, we test larval Drosophila for their cognitive abilities in three tasks that are characteristically more complex than those previously studied. Our data provide evidence for (i) conditioned inhibition, as has previously been reported for adult flies and honeybees. Unlike what is described for adult flies and honeybees, however, our data do not provide evidence for (ii) sensory preconditioning or (iii) second-order conditioning in Drosophila larvae. We discuss the methodological features of our experiments as well as four specific aspects of the organization of the larval brain that may explain why these two forms of learning are observed in adult flies and honeybees, but not in larval Drosophila.

Analysis of X-ray irradiation effects on the mortality values and hemolymph immune cell composition of Apis mellifera and its parasite, Varroa destructor

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.

Nosema apis and Nosema ceranae in Beehives of the Apulian Region of Italy: How Citizen Science Can Support Scientific Research

Nosema apis and Nosema ceranae are two well-known pathogens affecting the health of honeybees. To help understand how honeybee colonies are affected by these pathogens, the aim of this study was to analyze the impact of Nosema ceranae and Nosema apis in hives in the Apulian Region through a Citizen Science approach. First, a form about the health status of the beehives was filled out by beekeepers. After an inspection visit to confirm the signs observed by beekeepers, adult honeybee samples collected from beehives in four Apulian provinces (Taranto, Bari, Foggia and Brindisi) were subjected to light microscopy investigation for the detection of Nosema spp. spores and to molecular analysis using species-specific primers for the discrimination of Nosema apis spores from those of Nosema ceranae. Among the forty-eight samples, thirty-six tested positive for Nosema ceranae, and one sample tested positive for Nosema apis. The forms filled out by beekeepers revealed that only 5/36 beehives that tested positive for Nosema ceranae showed signs of depopulation and reduced honey production, while 19/36 had only low honey yield. This study provides data on Nosema apis and Nosema ceranae prevalence in Italy and correlates the presence of these intestinal pathogens with the most important problems encountered by local beekeepers.