Apis Mellifera Research Articles

Morphological characterization of spermatozoa in Apis mellifera and the effect of processing and semen storage

The aim of this study was to establish a standardized approach for evaluating sperm morphology in the honey bee and to investigate the impact of processing techniques and semen storage on the incidence of sperm morphoanomalies. The first experiment was designed to characterize the sperm morphoanomalies in unaltered honey bee semen, examining if they are compatible with sperm motility and viability and determining their occurrence in the ejaculates of mature and healthy honey bee drones. The different forms of sperm morphoanomalies were described in detail. In the morphological analysis of fresh drone ejaculates, 7.77% of spermatozoa showed abnormal morphology on average, with 2.20%, 4.75% and 0.81% of head, tail and multiple defects, respectively. The method also allowed us to determine the status of acrosome integrity. The second experiment was designed to assess the impact of smearing and air-drying on the occurrence of sperm morphoanomalies, showing a significant increase in the incidence of both head and tail sperm defects when comparing to wet samples. In the third and fourth experiments, the effect of semen storage at room temperature and semen cryopreservation, respectively, on the occurrence of sperm morphoanomalies were investigated. The preservation of semen at 22 °C led to a significant increase in spermatozoa with coiled tails after day 1, whereas the remaining anomalies did not exhibit significant variations over time. The freezing-thawing process showed a more pronounced effect on the incidence of morphological defects, with an increase in the percentage of spermatozoa with deformed heads and coiled tails.

Reliable molecular detection of small hive beetles

Invasive species require adequate reliable detection methods to mitigate their further spread and impact. However, the reliability of molecular detection methods is often hampered by both false positives (Error type I) and false negatives (Error type II). At present, the reliability of the four published molecular detection methods for small hive beetles (SHB), Aethina tumida, has not been rigorously evaluated considering their extensive genetic diversity. Here, we performed intra- and interlaboratory comparisons of the four available methods using SHB samples representing 78 regions from 27 countries on five continents, beetles from the same genus (Aethina concolor, A. inconspicua, A. flavicollis and A. major), as well as western honey bees, Apis mellifera, and ectoparasitic mites Varroa destructor. The data show that the Idrissou et al. (2018) and Li et al. (2018) methods avoid both false positives and false negatives probably due to lower sensitivity to nucleotide mismatches on the primer and probe’s target sequences. Further, the Li et al. (2018) method can be considered more sensitive because the fluorescent amplification curve crosses the threshold at lower Cq values compared to the Idrissou et al. (2018) one. In light of our data, the Li et al. (2018) method is the most reliable molecular diagnostic tool for SHB. We therefore recommend using this method as it will contribute to management efforts of this invasive species.

Viral Diversity in Mixed Tree Fruit Production Systems Determined through Bee-Mediated Pollen Collection

Commercially cultivated Prunus species are commonly grown in adjacent or mixed orchards and can be infected with unique or commonly shared viruses. Apple (Malus domestica), another member of the Rosacea and distantly related to Prunus, can share the same growing regions and common pathogens. Pollen can be a major route for virus transmission, and analysis of the pollen virome in tree fruit orchards can provide insights into these virus pathogen complexes from mixed production sites. Commercial honey bee (Apis mellifera) pollination is essential for improved fruit sets and yields in tree fruit production systems. To better understand the pollen-associated virome in tree fruits, metagenomics-based detection of plant viruses was employed on bee and pollen samples collected at four time points during the peak bloom period of apricot, cherry, peach, and apple trees at one orchard site. Twenty-one unique viruses were detected in samples collected during tree fruit blooms, including prune dwarf virus (PDV) and prunus necrotic ringspot virus (PNRSV) (Genus Ilarvirus, family Bromoviridae), Secoviridae family members tomato necrotic ringspot virus (genus Nepovirus), tobacco necrotic ringspot virus (genus Nepovirus), prunus virus F (genus Fabavirus), and Betaflexiviridae family member cherry virus A (CVA; genus Capillovirus). Viruses were also identified in composite leaf and flower samples to compare the pollen virome with the virome associated with vegetative tissues. At all four time points, a greater diversity of viruses was detected in the bee and pollen samples. Finally, the nucleotide sequence diversity of the coat protein regions of CVA, PDV, and PNRSV was profiled from this site, demonstrating a wide range of sequence diversity in pollen samples from this site. These results demonstrate the benefits of area-wide monitoring through bee pollination activities and provide new insights into the diversity of viruses in tree fruit pollination ecosystems.

Addressing bee diversity through active learning methodologies enhances knowledge retention in an environmental education project

The Brazilian ecosystems harbor a significant portion of global bee diversity. Despite their magnitude as pollinators, Brazilian native bees are mainly unknown. Considering the importance of scientific literacy in fostering environmental responsibility, we analyzed the BeeDiversity project’s impact on the scientific knowledge of eight to nine-year-old students. This study outlines the challenges of implementing the BeeDiversity project as a curricular component, including live bee observation and active learning methodologies. Students’ knowledge about bees significantly improved after participating in the project. Although the students mastered the concept of the ecological role of bees as pollinators, they were unaware that this ecosystem service is operated mainly by solitary bees. Learning retention was confirmed after five months. Active learning proved more effective than traditional methods. Handling a live hive and learning about solitary bees’ pollination roles generated high enthusiasm and engagement. Therefore, focusing on educational strategies addressing bee diversity beyond the typical honeybee (<i>Apis mellifera</i>) might effectively stimulate a broad environmental consciousness.

Invasive Ant Detection: Evaluating Honeybee Learning and Discrimination Abilities for Detecting Solenopsis invicta Odor.

Invasive red imported fire ants (Solenopsis invicta) create a serious threat to public safety, agriculture, biodiversity, and the local economy, necessitating early detection and surveillance, which are currently time-consuming and dependent on the inspector's expertise. This study marks an initial investigation into the potential of honeybees (Apis mellifera) to detect and discriminate the odor of S. invicta through the olfactory conditioning of proboscis extension responses. Deceased S. invicta were used as conditioned stimuli to ensure relevance to non-infested areas. The results showed that the bees rapidly learned to respond to deceased ant odors, with response levels significantly increasing at higher odor intensities. Bees exhibited generalization across the odors of 25 minor workers, 21 median workers, 1 major worker, and 1 female alate. When conditioned with deceased ant odors, bees effectively recognized live ants, particularly when trained on a single minor worker. Discrimination abilities varied by species and were higher when S. invicta was paired with Polyrhachis dives and Nylanderia yaeyamensis, and lower with S. geminata, Pheidole rabo, and Pheidole fervens. Notably, discrimination improved significantly with the application of latent inhibition. These findings suggest that trained honeybees have the potential to detect S. invicta. Further refinement of this approach could enhance its effectiveness for detection and surveillance.

Chronic oral toxicity protocol for adult solitary bees (Osmia bicornis L.): Reduced survival under long-term exposure to a “bee-safe” insecticide

Pollinators are essential for crop productivity. Yet, in agricultural areas, they may be threatened by pesticide exposure. Current pesticide risk assessments predominantly focus on honey bees, with a lack of standardized protocols for solitary bees. This study addresses this gap by developing a long-term oral exposure protocol tailored for O. bicornis. We conducted initial trials to determine optimal container sizes and feeding methods, ensuring high survival rates and accurate syrup consumption measurements. A validation test involving five laboratories was then conducted with the insecticide Flupyradifurone (FPF). Control mortality thresholds were set at ≤ 15% at 10 days. Three laboratories achieved ≤10%, demonstrating the protocol's effectiveness in maintaining healthy test populations. The seasonal timing of experiments influenced control mortality, underscoring the importance of aligning tests with the natural flight period of the population used. Our findings revealed dose-dependent effects of FPF on syrup consumption, showing stimulatory effects at lower concentrations and inhibitory effects at higher ones. The 10-day median lethal daily dose (LDD50) of FPF for O. bicornis (531.92 ng/bee/day) was 3.4-fold lower than that reported for Apis mellifera (1830 ng/bee/day), indicating Osmia's higher susceptibility. Unlike other insecticides, FPF did not exhibit time-reinforced toxicity. This study introduces a robust protocol for chronic pesticide exposure in solitary bees, addressing a critical gap in current risk assessment. Based on its low risk to honey bees and bumblebees, FPF is approved for application during flowering. However, our results suggest that it may threaten Osmia populations under realistic field conditions. Our findings underscore the need for comparative toxicity studies to ensure comprehensive protection of all pollinators and the importance of accounting for long term exposure scenarios in risk assessment. By enhancing our understanding of chronic pesticide effects in solitary bees, our study should contribute to the development of more effective conservation strategies and sustainable agricultural practices.

Mud Hive: An Eco-friendly Approach for Apis mellifera

The stationary mud hive, designed as a non-transportable alternative to the BIS wooden hives, adheres to the same frame dimensions. A site with direct sunlight is chosen, and the hive is built using a mud mixture, iron planks, and standard Indian bricks (7.5 × 3.54 × 3.54 inches). A brick with a bee entry gate (6 × 2 inches) is made. The entire structure is then layered with mud, providing effective thermal insulation. After drying and sintering for strength, it holds up to 10 BIS frames. This traditional construction approach offers a reliable and environmentally sustainable alternative to modern beekeeping methods.

The Effect of Hive Type on Colony Homeostasis and Performance in the Honey Bee (Apis mellifera).

The colonies of honey bees are mostly sessile organisms. Consequently, the type of nest boxes that beekeepers provide to their bees should impact a colony's ability to maintain homeostasis, which is a key determinant of performance and fitness. Here, we used European honey bees (Apis mellifera) and provided them with two hive setups widely used and known as Langstroth and Warré. We compared colony performance in a Mediterranean climate for five months from late spring to early autumn, which covered the most active time of bees and included periods of heat and drought. We found that irrespective of hive type or season, honey bees kept hive temperature and humidity within a remarkably narrow range. Nevertheless, the hive type impacted the daily fluctuations in temperature and humidity. In Warré hives, where bees have more autonomy to build and maintain their combs, we found that bees were able to reduce daily fluctuations in temperature and humidity and kept both measures closer to the overall average. This increase in colony homeostasis found in Warré hives negatively correlated with other hive performance indicators, such as immunocompetence. We conclude that different hive types affect key areas, such as the central part of the colony with frames of developing brood or the queen, which are the most susceptible individuals. This implies that climatic changes resulting in extreme weather events are expected to impact colony performance and fitness, especially in non-managed honey bees that are limited by available nesting sites. For managed bees, adaptations to existing hive setups could be provided to help bees minimize the effects of abiotic stress.

Complete genome sequence of deformed wing virus isolated from honeybees (Apis mellifera L.) in Iraq.

Here, we report the complete genome sequence of deformed wing virus (DWV) isolated from honeybees (Apis mellifera L.) in Iraq, named the Tikrit isolate. The phylogeny revealed that the Tikrit isolate has a close relationship to the Iraqi isolate DWV-Iraq-2023 and related to isolates from France, the United Kingdom, and Israel.

In vitro cytotoxicity assessment of biosynthesized Apis mellifera bee venom nanoparticles (BVNPs) against MCF-7 breast cancer cell lines

In this work, we reported the synthesis of honey bee (Apis mellifera) venom-derived nanoparticles via a hydrothermal method. This method not only ensures the preservation of the bee venom’s bioactive components but also enhances their potential stability, thus broadening the scope for their applications in the biomedicinal field. The synthesis method started with the homogenization suspension of bee venom, followed by its hydrothermal process to synthesize bee venom nanoparticles (BVNPs). The successful synthesis of BVNPs was characterized using various characteristic techniques such as Ultraviolet–visible (UV–Vis) spectroscopy, Fourier Transforms Infrared (FTIR) Spectroscopy, Zeta Potential (ZP), Liquid Chromatography-Mass Spectrometry (LCMS), and Transmission Electron Microscopy (TEM). The synthesis of BVNPs through biosynthesis is shown by the visible violet-brown color development at 347 nm by UV–Vis spectroscopy. FTIR analysis revealed the presence of several functional groups in the BVNPs, including alcohols (–OH), phenols (C6H5–), carboxylic acids (–COOH), amines (–NH2, –NH–), aldehydes (–CHO), ketones (–CO–), nitriles (–CN), amides (–CO–N–), imines (–CNH–), esters (–COO–), and polysaccharides. These functional groups, as confirmed by their specific stretching and bending vibrational modes, contribute to the diverse biological activities of BVNPs, including cytotoxicity against MCF-7 breast cancer cells. The ZP of the BVNPs indicated good colloidal stability at − 45 mV. LCMS analysis confirmed the presence of major bioactive molecules, including melittin & apamin and TEM analysis shows the BVNPs exhibited a quasi-spherical shape with good dispersion, the average size was approximately 25 nm, with some being smaller (quantum dots) and interplanar spacing of 0.236 nm indicated a highly ordered crystalline structure. Moreover, the anticancer efficacy of the BVNPs was ascertained through in vitro assays against MCF-7 breast cancer cells, showing a dose-dependent cytotoxic effect. The findings of this study underscore the viability of hydrothermal synthesis in producing biologically active and structurally stable BVNPs, with a significant potential for anticancer activities.Graphical

A test method for assessing chronic oral toxicity of a pesticide to solitary nesting orchard bees, Osmia spp. (Hymenoptera: Megachilidae).

Orchard bees of the genus Osmia Panzerare important pollinators of fruit trees in various regions of the world, with some species commercially available in the United States and Europe. In addition to their pollination services, Osmia lignaria, Osmia cornifrons, Osmia bicornis, and Osmia cornuta have been identified as potential model species for solitary bees in pesticide risk assessment and have been used for the development of new methods to test acute lethal effects via contact and oral routes of exposure. Our goal was to expand the available methodology to characterize the toxicity of pesticides for these solitary bees through a chronic oral test for adult bees. Chronic oral toxicity of pesticides to orchard bees has been reported, but methods differ among research groups. In our study, O. lignaria, O. cornifrons, O. bicornis, and O. cornuta female bees had access to sucrose solution ad libitum in separate, species-specific 10-day tests. Mean body mass, mean daily consumption, and survival differed among the studied bee species. The dose-response test design was validated with dimethoate, a reference toxic compound, and chronic toxicity endpoints were estimated for the 4 Osmia species. The median lethal daily doses normalized by weight for O. lignaria, O. bicornis, O. cornuta, and O. cornifrons were within the same order of magnitude at 0.23, 0.26, 0.49, and 0.61 µg dimethoate/g bee/day, respectively. The methodology described here was aligned as much as possible with the available honey bee and bumble bee standard methods to facilitate the comparison of chronic toxicity profiles among bee species.

Virulence-linked adhesin drives mutualist colonization of the bee gut via biofilm formation.

Bacterial biofilms are stable multicellular structures that can enable long term host association. Yet, the role of biofilms in supporting gut mutualism is still not fully understood. Here, we investigate Snodgrassella alvi , a beneficial bacterial symbiont of honey bees, and find that biofilm formation is required for its colonization of the bee gut. We constructed fifteen S. alvi mutants containing knockouts of genes known to promote colonization with putative roles in biofilm formation. Genes required for colonization included staA and staB , encoding trimeric autotransporter adhesins (TAAs) and mltA , encoding a lytic transglycosylase. Intriguingly, TAAs are considered virulence factors in pathogens but support mutualism by the symbiont S. alvi. In vitro , biofilm formation was reduced in Δ staB cells and abolished in the other two mutants. Loss of staA also reduced auto-aggregation and cell-cell connections. Based on structural predictions, StaA/B are massive (>300 nm) TAAs with many repeats in their stalk regions. Further, we find that StaA/B are conserved across Snodgrassella species, suggesting that StaA/B-dependent colonization is characteristic of this symbiont lineage. Finally, staA deletion increases sensitivity to bactericidal antimicrobials, suggesting that the biofilm indirectly buffers against antibiotic stress. In all, the inability of two biofilm-deficient strains (Δ staA and Δ mltA ) to effectively mono-colonize bees indicates that S. alvi biofilm formation is required for colonization of the bee gut. We envision the bee gut system as a genetically tractable model for studying the physical basis of biofilm-mutualist-gut interactions.

Study of the chemical composition of the essential oil derived from Thymus lancéolatus desf, an endemic aromatic plant native from Algeria and evaluation of its efficacy against Paenibacillus larvae and Ascosphaera Apis, two diseases affecting the honeybees, Apis mellifera

The essential oil and hydrolate of Thymus lancéolatus desf, an endemic aromatic plant native to the mountains of northwestern Algeria, were tested in vitro for their antimicrobial activity against the bacteria Paenibacillus larvae, the causative agent of American foulbrood, and the fungi Ascosphaera apis, the responsible agent of chalkbrood disease in honeybees, Apis mellifera. The oil was extracted from various aerial parts of the plant using steam distillation. It shows high efficacy against both diseases, while its hydrolat demonstrates high efficacy against A.apis alone. The lethal doses (LD100) of the fungi were estimated at 0.034% and 25.30% for the essential oil and the hydrolat, respectively. The estimated concentration of the DL100 bacteria was 17.37 ul/Disc of EO. The hydrolate had no effect on P. larvae. This exceptional double efficacy of T.lancéolatus desf EO has been attributed to its particular active compound, Durenol, which constitutes 34.8% of the oil and the high quantities of its precursors (γ-Terpinene (16.71%) and Sylvestrene (15.17%) contribute to its effectiveness. Also, the abcence of acetate compound further enhances its efficacy. EO of Thyme was proven to be the most effective essential oil in vitro against P. larvae and A. apis, making it an excellent natural alternative to antibiotics.

Bioactive compounds enrichment in rabbit doe’s diet pre-and during pregnancy improves productive and reproductive performance and cost-effectiveness under hot climates

This study aimed to investigate the effect of diet supplementation with a symbiotic (SY), bee pollen (BP), honey bee (HB), date palm pollen (DPP) and their mixture (MIX) on female rabbit productive and reproductive performances under desert hot climates. Seventy-two Californian does of 5 months age and average body weight of 3250 ± 78.2 g were randomly allotted into six groups, each of 12 does. All does orally receive 3 ml distilled water for 10 days before mating and 28 days during pregnancy. Treatments were repeated for four consecutive parities. The first group served as control (C) given distilled water only, however the second, third, fourth and fifth groups were supplemented with 3 ml distilled water containing 0.2 ml SY, 200 mg DPP, 200 mg BP, 0.2 ml HB/doe per day, respectively. While, the sixth group does were given all previous ingredients (MIX). Sexual receptivity rate, fertility rate, kindling rate, and newborn traits were recorded. Also, maternal feed intake, feed conversion ratio, and digestibility coefficients of nutrients were recorded. Does in all groups were artificially inseminated with 0.5 ml of fresh heterospermic semen of 15 fertile bucks extended in Tris at 806–1006 sperm/ml. Treatment increased maternal body weight and daily gain with highest values (P < 0.05) in BP, SY, and MIX does. All treatments enhanced feed intake and feed conversion ratio (FCR) compared with control. Number services per conception decreased (P < 0.01), while litter size and weight and survival at birth and weaning increased (P < 0.01) in treated than control does. Treated does produced more milk than control. Digestibility coefficients of all nutrients were improved (P < 0.01) in treated does. In conclusion, supporting rabbit does pre- and during pregnancy with diets supplemented with a mixture of honey bee, date palm pollen, bee pollen, and synbiotic improves the productive and reproductive performances of rabbit does and their offspring.

Foraging distance distributions reveal how honeybee waggle dance recruitment varies with landscape

Honeybee (Apis mellifera) colonies use a unique collective foraging system, the waggle dance, to communicate and process the location of resources. Here, we present a means to quantify the effect of recruitment on colony forager allocation across the landscape by simply observing the waggle dance on the dancefloor. We show first, through a theoretical model, that recruitment leaves a characteristic imprint on the distance distribution of foraging sites that a colony visits, which varies according to the proportion of trips driven by individual search. Next, we fit this model to the real-world empirical distance distribution of forage sites visited by 20 honeybee colonies in urban and rural landscapes across South East England, obtained via dance decoding. We show that there is considerable variation in the use of dancing information in colony foraging, particularly in agri-rural landscapes. In our dataset, reliance on dancing increases as arable land gives way to built-up areas, suggesting that dancing may have the greatest impact on colony foraging in the complex and heterogeneous landscapes of forage-rich urban areas. Our model provides a tool to assess the relevance of this extraordinary behaviour across modern anthropogenic landscape types.

Significance of honeybee pollination in increasing seed yield of Trifolium alexandrinum (Fabales: Fabaceae)and its impact on economic sustainability of smallholder farmers.

A major limitation to producing high seed yields in berseem clover (Trifolium alexandrinum L.) is failure to set seed, predominantly due to lack of pollination. Despite the importance of berseem clover as a leading forage legume, the contribution of pollinators to seed set is scarce. In Pakistan, the honeybee population is declining mainly because of the extensive use of neonicotinoid pesticides and habitat fragmentation. This, combined with the region's harsh environment and the use of inferior, locally bred genotypes, has resulted in low seed yields. Insufficient seed availability leads to limited forage supply, resulting in poor livestock nutrition, which subsequently impacts livestock health and productivity, and reduces farmers' income. The present study estimated the seed production of 3 berseem clover genotypes resulting from honeybees [Apis mellifera L. (Hymenoptera: Apidae)] pollination in 2 growing seasons (2012-2014) in the central Punjab region of Pakistan. Experiments had 2 pollination treatments (open pollination and honeybee inclusion) and 3 seed genotypes, viz. farmer own-saved, market, and the improved variety cultivars. For both growing seasons, honeybee pollination resulted in significant increases in seed yields ranging from 35% to 67%, regardless of seed genotype. With the exception of the number of seed heads/m2, all seed yield parameters also increased significantly in response to honeybee inclusion. The combination of improved variety and honeybee inclusion resulted in the production of a maximum number of seeds per head (45.3), 1,000-seed weight (3.7g), and estimated seed yield (375.5kg/ha). In addition, the increase in estimated net income of seed ranged from PKR 82,485 Rs/ha (US$844/ha) to PKR 168,975 Rs/ha (US$1728/ha) with the use of honeybees as an insect pollinator across all the seed genotypes. Honeybee pollination has broader implications for mixed farming systems by playing a key role in preserving biodiversity and promoting sustainable agriculture. It also enhances the quality and quantity of berseem crops by increasing the production of high-quality seeds and forage leading to improved livestock productivity and family food security which strengthens the economic resilience of rural communities.

Comparative Transcriptome Analysis of Cold Tolerance Mechanism in Honeybees (Apis mellifera sinisxinyuan).

Honeybees are important pollinators worldwide that are closely related to agricultural production and ecological balance. The biological activities and geographical distribution of honeybees are strongly influenced by temperature. However, there is not much research on the cold tolerance of honeybees. The Apis mellifera sinisxinyuan, a kind of western honeybee, exhibits strong cold hardiness. Here, we determined that short-term temperature treatment could regulate the honeybee's cold tolerance ability by measuring the supercooling point of A. m. sinisxinyuan treated with different temperatures. Transcriptome data were analyzed between the treated and untreated honeybees. A total of 189 differentially expressed genes were identified. Among them, Abra, Pla1, rGC, Hr38, and Maf were differentially expressed in all comparisons. GO and KEGG analysis showed that the DEGs were enriched in molecular functions related to disease, signal transduction, metabolism, and the endocrine system's function. The main components involved were ribosomes, nucleosomes, proteases, and phosphokinases, among others. This study explored the formation and regulation mechanism of cold tolerance in honeybees, not only providing a theoretical basis for cultivating honeybees with excellent traits but also promoting research and practice on insect stress tolerance.

Nosema ceranae infection reduces the fat body lipid reserves in the honeybee Apis mellifera

Nosema ceranae is an intestinal parasite frequently found in Apis mellifera colonies. This parasite belongs to Microsporidia, a group of obligate intracellular parasites known to be strongly dependent on their host for energy and resources. Previous studies have shown that N. ceranae could alter several metabolic pathways, including those involved in the nutrient storage. To explore the impact of N. ceranae on the fat body reserves, newly emerged summer bees were experimentally infected, and we measured (1) the lipid percentage of the abdominal fat body at 2-, 7- and 14-days post-inoculation (p.i.) using diethyl ether lipid extraction, (2) the triglyceride and protein concentrations by spectrophotometric assay methods, and (3) the amount of intracellular lipid droplets in trophocytes at 14- and 21-days p.i. using Nile Red staining. Comparing the three methods used to evaluate lipid stores, our data revealed that Nile Red staining seemed to be the simplest, fastest and reliable method. Our results first revealed that the percentage of fat body lipids significantly decreased in infected bees at D14 p.i. The protein stores did not seem to be affected by the infection, while triglyceride concentration was reduced by 30% and lipid droplet amount by 50% at D14 p.i. Finally, a similar decrease in lipid droplet reserves in response to N. ceranae infection was observed in bees collected in fall.

Honey bee viruses in the yellow-legged hornet Vespa velutina (Lepelieter 1836): Prevalence, loads, and detection of replicative DWV and LSV forms

Apiaries in Galicia, northwestern Spain, are currently facing the invasive alien species Vespa velutina, which is well established in the region. The pressure on honey bee colonies is high, resulting in both economic and ecological losses. Honey bee colonies also face the challenge of viruses, which are becoming increasingly diverse. In recent years, honey bee viruses have been spreading across taxonomic groups beyond Apoidea, infecting the Vespoidea superfamily. This cross-species spillover has raised concerns in the scientific community due to the potential risk of viruses spreading in ecosystems. Currently, there is a lack of knowledge on this topic, and further research is needed to address this issue. This study employed qPCR and sequencing to investigate the prevalence, loads, and presence of replicative forms of important honey bee viruses in V. velutina individuals collected from 11 apiaries in Galicia. All V. velutina individuals tested positive for DWV, BQCV, AKI complex (ABPV, KBV, and IAPV), or LSV but not for CBPV. DWV showed the highest prevalence (97.0 %) and loads, with both DWV-A (67.4 %) and DWV-B (32.6 %) being detected. The AKI complex (46.3 %) and LSV (43.3 %) were also common, whereas BQCV (11.9 %) was rarer. LSV is detected for the first time in V. velutina. LSV-2 was the dominant strain (82.1 %), and two less frequent (17.9 %) unknown strains were also detected. All 44 screened V. velutina samples carried the replicative form of DWV, and six of these also carried the replicative form of LSV, raising for the first time the possibility of co-infection in the hornet. The detection of honey bee viruses in V. velutina, and the ability of these viruses to spread to other species, may indicate a potential risk of spillover in the apiaries.

Molecular characterization and expression pattern analysis of Sirtuin family genes in the eastern honeybee, Apis cerana

Sirtuin proteins are a family of evolutionarily conserved NAD+-dependent deacetylase enzymes which play crucial roles in regulating insect metabolism, maintaining genome stability, and lifespan. However, there is no report on the genome-wide analysis of the Sirtuin gene family in the eastern honeybee, Apis cerana. Here, 6 Sirtuin genes (AcSirt1, AcSirt2, AcSirt4 ~ AcSirt7) were obtained in the A. cerana genome, which were located on 6 different chromosomes and were all single-copy genes. Gene structure analysis revealed that these 6 genes were all multi-exon genes. The Sirtuin family genes contained 9 conserved motifs and 5 conserved domains, which exhibited typical Sirtuin protein structural characteristics. The phylogenetic analysis of the Sirtuin family genes indicates that A. cerana lacked the Sirt3 gene, and the 6 Sirtuin encoding genes could be clustered into 5 groups and 6 branches, showing high homology with the Apis mellifera and the Apis dorsata. The results of the tissue and period expression profiles by qRT-PCR indicated that the 6 Sirtuin family genes showed transcriptional activity in different tissues of worker bees, and the gene expression level reached the peak in the 4-day-old larvae and pupa stages. It was speculated that Sirtuin genes expression level was affected by fasting behavior, caloric restriction and organ differentiation during different developmental stages of A. cerana. Meanwhile, the differential expression patterns of Sirtuin genes in various tissues further demonstrated the diversity and complexity of their functions, providing a basis for target selection in future studies on the functionality of Sirtuin genes.