Mellifera Workers Research Articles

Morphometric Characterization of Apis mellifera (Italian Honey Bee) Across Different Agroclimatic Zones of Kumaun Region of Uttarakhand, India

This study investigated the morphometry of Apis mellifera across three diverse locations in Uttarakhand, spanning the Tarai irrigated (Pantnagar), Bhabar Irrigated (Nainital) and High hills (Almora), to explore regional adaptations and environmental influences. Significant regional differences were observed, with populations from high hills exhibiting larger dimensions, whereas populations from Tarai irrigated showed smaller size. The morphological characteristics of Apis mellifera worker bees were analyzed. A total of 13 morphological characters were measured using a stereo zoom microscope in combination with VUE software. The results suggest that morphometric variations can potentially distinguish intraspecific groups of honeybee colonies, identifying them as distinct ecotypes. Morphometric analysis of wings and body partsprovides valuable insights into honeybee biodiversity studies To facilitate species identification based on morphometric traits, Apis mellifera samples were collected. 40 worker bee samples were analyzed, and 13 morphometric characteristics were measured in the entomology laboratory at G.B. Pant University of Agriculture and Technology (GBPUA&T). The biometric data were analyzed by calculating the mean, range, standard error, and coefficient of variation.

Dietary dopamine supplementation has no effect on ovary activity in queen-less or queen-right honeybee (Apis mellifera) workers

Eusociality in honeybees (Apis mellifera) is characterised by a reproductive division of labour, where the queen monopolises reproduction while worker reproduction is constrained. This constraint is mediated by queen mandibular pheromone (QMP), which inhibits worker ovary development through Notch signalling and possibly oocyte apoptosis. Dopamine has been implicated in regulating reproduction in worker honeybees, with prior studies suggesting that dietary dopamine enhances ovary activity in queen-less workers. This study aimed to test whether dopamine supplementation could overcome QMP-mediated reproductive constraint in worker honeybees. Using caged honeybee experiments, we administered dopamine and its precursor L-dopa at a range of concentrations, both in the presence and absence of QMP. Results showed that neither dopamine nor L-dopa supplementation affected ovary development, survival rates, or food intake, which contrasts with a previous study. These findings suggest that dopamine may not be a major factor in QMP-mediated reproductive inhibition. Instead, we suggest that the multifaceted nature of QMP’s components together with the complexity of neuroendocrine signalling makes it likely that multiple redundant mechanisms regulate worker reproduction. Future research should investigate the interplay between nutrition, dopamine and QMP components to fully understand the regulation of ovary activation in honeybee workers.

LC-MS profiling reveals metabolic dynamics in Apis mellifera worker bee larvae–pupae transition

LC-MS profiling reveals metabolic dynamics in Apis mellifera worker bee larvae–pupae transition

Stressdrives premature hive exiting behavior that leads to death in young honey bee (Apis mellifera) workers.

The Western honey bee, Apis mellifera, is an economically important pollinator, as well as a tractable species for studying the behavioral intricacies of eusociality. Honey bees are currently being challenged by multiple biotic and environmental stressors, many of which act concomitantly to affect colony health and productivity. For instance, developmental stress can lead workers to become precocious foragers and to leave the hive prematurely. Precocious foragers have decreased flight time and lower foraging efficiency, which can ultimately lower colony productivity and even lead to colony collapse. In this study, we tested the hypothesis that stress during pupal development can cause young workers to exit the hive prematurely before they are physically able to fly. This premature exiting behavior results in death outside the hive soon thereafter. To determine how various stressors may lead bees to perform this behavior, we subjected workers during the last pupal stage to either cold stress (26°C for 24h), heat stress (39°C for 24h), or Varroa destructor mite parasitization, and compared the rate of premature hive exits between stressed bees and their respective control counterparts. Upon emergence, we individually tagged focal bees in all treatment groups and introduced them to a common observation hive. We then followed tagged bees over time and monitored their survivorship, as well as their likelihood of performing the premature hive exiting behavior. We also dissected the hypopharyngeal glands of all treatment and control bees sampled. We found that significantly more bees in all three treatment groups exited the hive prematurely compared to their control counterparts. Bees in all treatment groups also had significantly smaller hypopharyngeal glands than control bees. Our results suggest that premature hive exiting behavior is driven by stress and is potentially a form of accelerated age polyethism that leads to premature death.

Azoxystrobin Exposure Impacts on Development Status and Physiological Responses of Worker Bees (Apis mellifera L.) from Larval to Pupal Stages.

Honeybee larvae and pupae form the cornerstone of colony survival, development, and reproduction. Azoxystrobin is an effective strobilurin fungicide that is applied during the flowering stage for controlling plant pathogens. The contaminated nectar and pollen resulting from its application are collected by forager bees and impact the health of honeybee larvae and pupae. The current study evaluated the survival, development, and physiological effects of azoxystrobin exposure on the larvae and pupae of Apis mellifera worker bees. The field-recommended concentrations of azoxystrobin were found to suppress the survival indices and lifespan in the larval as well as pupal stages; moreover, the rates of the survival and pupation of larvae as well as the body weights of the pupae and newly-emerged adult bees were significantly reduced upon long-term exposure to azoxystrobin. In addition, azoxystrobin ingestion induced changes in the expression of genes critical for the development, immunity, and nutrient metabolism of larvae and pupae, although the expression profile of these genes differed between the larval and pupal stages. Results indicated the chronic toxicity of azoxystrobin on the growth and development of honeybee larvae and pupae, which would affect their sensitivity to pathogens and other external stresses during the development stage and the study will provide vital information regarding the pollination safety and rational use of pesticides.

Dedicated developmental programing for group-supporting behaviors in eusocial honeybees.

The evolutionary changes from solitary to eusocial living in vertebrates and invertebrates are associated with the diversification of social interactions and the development of queen and worker castes. Despite strong innate patterns, our understanding of the mechanisms manifesting these sophisticated behaviors is still rudimentary. Here, we show that doublesex (dsx) manifests group-supporting behaviors in the honeybee (Apis mellifera) worker caste. Computer-based individual behavioral tracking of worker bees with biallelic stop mutations in colonies revealed that the dsx gene is required for the rate and duration of group-supporting behavior that scales the relationship between bees and their work. General sensorimotor functions remained unaffected. Unexpectedly, unlike in other insects, the dsx gene is required for the neuronal wiring of the mushroom body in which the gene is spatially restricted expressed. Together, our study establishes dedicated programming for group-supporting behaviors and provides insight into the connection between development in the neuronal circuitry and behaviors regulating the formation of a eusocial society.

Tracking Varroa Parasitism Using Handheld Infrared Cameras: Is Eusocial Fever the Key?

The Varroa destructor is the most significant bee parasite and the greatest threat to bee health all around the world. Due to its hidden lifestyle, detection within the brood cell is only possible through invasive techniques. Enhancing detection methods is essential for advancing research on population dynamics, spread, selection efforts, and control methodologies against the mite. In our study, we employed infrared imaging to measure the thermal differences in parasite and intact Apis mellifera worker broods. Experiments were conducted over two years at the MATE Kaposvár Campus in Hungary involving five beehives in 2022 and five beehives in 2023. A FLIR E5-XT WIFI handheld infrared camera was used to create a heat map of capped brood frames. Our results indicate that the resolution of these cameras is sufficient to provide detailed IR images of a bee colony, making them suitable to detect temperature differences in intact and Varroa parasitized capped brood cells. Mite parasitism causes a time-dependent and sustained temperature increase in developing bee pupae, observable regardless of mite number. Our work demonstrates two different heating patterns: hotspot heating and heating cells that are responsible for the elevated temperature of the Varroa-infested cells as a social fever response by the worker bees. Based on our results, future research combined with AI-based image evaluation software could offer beekeepers and researchers practical and valuable tools for high-throughput, non-invasive Varroa detection in the field.

Sublethal Effects Induced by a Cyflumetofen Formulation on Honeybee Apis mellifera L. Workers: Assessment of Midgut, Hypopharyngeal Glands, and Fat Body Integrity.

Worldwide, both cultivated and wild plants are pollinated by the honey bee, Apis mellifera. Bee numbers are declining as a result of a variety of factors, including increased pesticide use. Cyflumetofen controls pest mites in some plantations pollinated by bees, which may be contaminated with residual sublethal concentrations of this pesticide, in nectar and pollen. We evaluated the effects of a sublethal concentration of a cyflumetofen formulation on the midgut, hypopharyngeal gland, and fat body of A. mellifera workers orally exposed for 72 h or 10 days. The midgut epithelium of treated bees presented digestive cells with cytoplasm vacuoles and some cell fragmentation, indicating autophagy and cell death. After being exposed to the cyflumetofen formulation for 72 h, the midgut showed a higher injury rate than the control bees, but after 10 days, the organs had recovered. In the hypopharyngeal gland of treated bees, the end apparatus was filled with secretion, suggesting that the acaricide interferes with the secretory regulation of this gland. Histochemical tests revealed differences in the treated bees in both exposure periods in the midgut and hypopharyngeal glands. The acaricide caused cytotoxic effects on the midgut digestive cells, with apical protrusions, plasma membrane rupture, and several vacuoles in the cytoplasm, features of cell degeneration. In the hypopharyngeal glands of the treated bees, the secretory cells presented small electron-dense and large electron-lucent secretory granules. The fat body cells had no changes in comparison with the control bees. In conclusion, the cyflumetofen formulation at sublethal concentrations causes damage to the midgut and the hypopharyngeal glands of honey bee, which may compromise the functions of these organs and colony fitness. Environ Toxicol Chem 2024;43:2455-2465. © 2024 SETAC.

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.

Impact of a worker bee thoracic ganglion RIC-3 variant on the actions of acetylcholine and neonicotinoids on nicotinic receptors in Apis mellifera.

A transmembrane thioredoxin (TMX3) enables the functional expression of insect nicotinic acetylcholine receptors (nAChRs) in Xenopus laevis oocytes, while co-factors RIC-3 and UNC-50 regulate the receptor expression level. RIC-3 (resistant to inhibitors of cholinesterase 3) has been shown to diversify by its differential mRNA splicing patterns. How such diversity influences neonicotinoid sensitivity of nAChRs of beneficial insect species remains poorly understood. We have identified a RIC-3 variant expressed most abundantly in the thoracic ganglia of honeybee (Apis mellifera) workers and investigated its effects on the functional expression and pharmacology of Amα1/Amα8/Amβ1 and Amα1/Amα2/Amα8/Amβ1 nAChRs expressed in X. laevis oocytes. The AmRIC-3 enhanced the response amplitude to the acetylcholine (ACh) of these A. mellifera nAChRs when its cRNAwas injected into oocytes at low concentrations but suppressed the ACh response amplitude at high concentrations. Co-expression of the AmRIC-3 had a minimal impact on the affinity of ACh, but changed the efficacy of imidacloprid and clothianidin, suggesting that the presence and the level of RIC-3 expression can affect the nAChR responses to ACh and neonicotinoids, depending on nAChR subunit composition in honeybees. © 2024 Society of Chemical Industry.

Acetylcholine and choline in honey bee (Apis mellifera) worker brood food are seasonal and age-dependent

Nursing honeybees produce brood food with millimolar concentrations of acetylcholine (ACh), which is synthesized through head gland secretions mixed with honey stomach contents. While we previously demonstrated the necessity of ACh for proper larval development, the dynamics of ACh levels throughout ontogenesis and their seasonal variations have remained unclear until now. Our HPLC analysis reveals dependencies of choline and ACh levels on larval development days (LDDs), influenced by seasonal (April–September) variations. Median ACh concentrations peak on LDD 2, declining significantly toward cell capping, while choline levels are lowest during the initial LDDs, rising markedly toward cell capping. Seasonal patterns show peak ACh levels from April to June and a low in August, paralleling choline's peak in July and low in August. This seasonality holds consistently across multiple years (2020–2022) and colonies, despite potential variations in colony performance and environmental conditions. Our analysis found no correlation between temperature, sunshine, precipitation, or favourable foraging days and ACh/choline levels, suggesting the involvement of additional factors. These findings underscore the seasonal fluctuation of ACh levels and its potential implications for the genetic programs governing winter bee development.

Action of dithiocarbimates salts on the honey bee and its pathogen Nosema ceranae

Apis mellifera, crucial pollinators for both native and cultivated plants, also yield various products such as honey, wax, royal jelly, and propolis, extensively utilized in the food, pharmaceuticals, and cosmetics industries. Nosema ceranae, a prevalent microsporidian worldwide, stands as a significant pathogen for A. mellifera, showing resistance to conventional antibiotics. Consequently, the exploration of novel compounds for N. ceranae control becomes imperative. Dithiocarbimate derivatives emerge as promising antifungal candidates under evaluation for combating various pathogens, particularly those affecting plants. This study assessed the toxicity profile of six dithiocarbimate derivatives on A. mellifera worker survival and N. ceranae pathogen. Among these, four compounds exhibited minimal bee mortality and proceeded to further evaluation against N. ceranae. In vitro assays demonstrated their inhibitory effects on spore germination. Remarkably, the most potent compound suppressed N. ceranae spores by 62% at a concentration of 20 µmol L−1in vivo. Thus, these dithiocarbimate derivatives represent promising new antifungal agents for combatting nosemosis in honey bee populations.

Effects of chronic oral exposure to insecticide teflubenzuron on the midgut of the honey bee Apis mellifera workers: histopathological insights into pesticide toxicity.

The honey bee Apis mellifera plays a significant role as a pollinator of native and cultivated plants, by increasing the productivity of several cultures, preserving the flora, and producing forest seeds. However, bee populations are declining worldwide, including A. mellifera, due to Colony Collapse Disorder, mainly resulting from the constant use of pesticides in the crops. Teflubenzuron is a physiological insecticide that belongs to the benzoylurea group, which inhibits chitin synthesis, the main component of the insect integument classified as safe for non-target insects, including bees. However, its effect on non-target organs of insects remains unknown. The midgut is the main organ of the digestive tract, which works in digestion and absorption and may be exposed to pesticides that contaminate food resources. The present work aimed to verify if the insecticide teflubenzuron is toxic and has histopathological effects on the midgut of A. mellifera adult workers. Workers exposed orally and chronically to the field-realistic concentration of teflubenzuron present 81.54% mortality. The epithelium of the midgut of these bees presents high vacuolization, spherocrystals, cell fragments released to the organ lumen, apocrine secretion, nuclear pyknosis, loss of cell-cell contact, and damage to regenerative cell nests and to the peritrophic matrix. These results indicate that the chitin synthesis-inhibiting insecticide teflubenzuron is toxic to A. mellifera after chronic oral exposure, at realistic field concentration, although it is classified as non-toxic to adult and non-target insects.

High-energy emulsification of Allium sativum essential oil boosts insecticidal activity against Planococcus citri with no risk to honeybees

AbstractThe ecotoxicological consequences of synthetic pesticides have encouraged stakeholders to search for eco-friendly pest control tools, like essential oils (EOs). Nano-delivery systems (nanoparticles and nano-emulsions) seem ideal for developing EO-based biopesticides, although production processes should be standardized and implemented. In this study, nano-emulsions loaded with a high amount of Allium sativum L. EO (15%) were developed using different mixed bottom-up/top-down processes. Garlic EO was chemically analyzed by gas chromatography-mass spectrometry (GC-MS) and formulations were physically characterized using Dynamic Light Scattering (DLS) apparatus. The insecticidal activity against Planococcus citri Risso (Hemiptera: Pseudococcidae) and selectivity toward Apis mellifera L. (Hymenoptera: Apidae) worker bees was evaluated. Garlic EO was mainly composed of sulphur components (96.3%), with diallyl disulphide and diallyl trisulphide as the most abundant compounds (37.26% and 28.15%, respectively). Top-down processes could produce stable nano-emulsions with droplet size in the nanometric range (< 200nm) and good polydispersity index (PDI < 0.2). In contrast, the bottom-up emulsion was unstable, and its droplet size was around 500nm after 24 hours. High-energy emulsification processes significantly increased the residual toxicity of garlic EO against 3rd instar P. citri nymphs, whereas the developed formulations were harmless to A. mellifera workers in topical application. This study confirmed that the production process significantly affected the physical properties and efficacy against target pests. The lack of adverse impact on honeybees denotated the potential of these formulations as bioinsecticides in organic and/or IPM programs, although further extended ecotoxicological studies are necessary.

Does consuming irradiated royal jelly affect Apis mellifera larvae development and survival to adulthood in vitro?

The purpose of this study was to determine if Apis mellifera could be reared in vitro on a diet containing royal jelly (RJ) irradiated at 25 kGy. Twelve-hour-old larvae were collected from fourteen colonies and fed a diet containing unirradiated RJ (refrigerated until use), irradiated RJ, or an irradiation control (RJ stored next to the irradiator for the time it takes to irradiate RJ). Statistically fewer larvae survived to adulthood when fed a diet containing irradiated RJ (70.6%) than when fed a diet containing unirradiated RJ (82.9%) or the irradiation control RJ (78.4%; F2,39 = 4.34; p < 0.001). Nevertheless, larval survival at day 8 and eclosion exceeded the U.S. Environmental Protection Agency’s requirements for controls in tier one toxicological studies. Feeding on irradiated RJ did not affect bee developmental time or weight at emergence. The data demonstrate that A. mellifera workers can be reared on a diet that includes irradiated RJ, but that additional diet refinements may be necessary to improve the survival of these individuals to levels experienced by larvae feeding on a diet containing unirradiated RJ.

Cytotoxic and genotoxic effects of a pendimethalin-based herbicide in Apis mellifera

Public concern about the effects of pesticides on non-target organisms has increased in the recent years. Nevertheless, there is a limited number of studies that address the actual toxic effects of herbicides on insects. This study investigated the side effects of herbicides on non-target organisms inhabiting agroecosystems and performing essential ecological and economic functions such as crop pollination. We analysed morphological alterations in the gut, Malpighian tubules and circulating haemocytes of Apis mellifera workers as markers of exposure effects. A commercial formulation of a pendimethalin-based herbicide (PND) was administered orally under laboratory conditions at a realistic concentration admitted in the field (330gL−1 of active ingredient., 4 L ha−1 for cereal and vegetable crops). The worker bees were exposed to a single application of PND for a period of one week, to simulate the exposure that can occur when foraging bees accidentally drink drops of contaminated water upon treatments. Histopathological analyses of the midgut, ileum and Malpighian tubules showed alterations over time (from 24 to 72 h after the beginning of exposure) such as loss of epithelial organisation, cellular vacuolisation and altered pyknotic nuclei as well as disruption of the peritrophic membrane over time. Semiquantitative analyses of the midgut showed a significant increase in the organ injury index 24 and 72 h after the initial exposure in PND-exposed bees compared to control bees. In addition, a change in positivity to Gram staining was observed in the midgut histological sections. A recovery of cytotoxic effects was observed one week after the initial exposure, which was favoured by the periodic renewal of the intestinal epithelium and the herbicide dissipation time. Cytochemical staining with Giemsa of haemocytes from PND-treated workers over 24 and 72 h showed significant nuclear alterations such as lobed or polymorphic nuclei and micronuclei compared to bees in the control group. These results show that the dose of PND used to protect crops from weeds can lead to significant cytotoxic and genotoxic effects in non-target organisms such as honey bees. In croplands, the sublethal effects on cell morphology can impair vital physiological processes such as nutrition, osmoregulation, and resistance to pathogens, contributing to the decline in biodiversity and abundance of species that play a prominent ecological role, such as pollinators.

Construction of a Full-Length Transcriptome of Western Honeybee Midgut Tissue and Improved Genome Annotation.

Honeybees are an indispensable pollinator in nature with pivotal ecological, economic, and scientific value. However, a full-length transcriptome for Apis mellifera, assembled with the advanced third-generation nanopore sequencing technology, has yet to be reported. Here, nanopore sequencing of the midgut tissues of uninoculated and Nosema ceranae-inoculated A. mellifera workers was conducted, and the full-length transcriptome was then constructed and annotated based on high-quality long reads. Next followed improvement of sequences and annotations of the current reference genome of A. mellifera. A total of 5,942,745 and 6,664,923 raw reads were produced from midguts of workers at 7 days post-inoculation (dpi) with N. ceranae and 10 dpi, while 7,100,161 and 6,506,665 raw reads were generated from the midguts of corresponding uninoculated workers. After strict quality control, 6,928,170, 6,353,066, 5,745,048, and 6,416,987 clean reads were obtained, with a length distribution ranging from 1 kb to 10 kb. Additionally, 16,824, 17,708, 15,744, and 18,246 full-length transcripts were respectively detected, including 28,019 nonredundant ones. Among these, 43,666, 30,945, 41,771, 26,442, and 24,532 full-length transcripts could be annotated to the Nr, KOG, eggNOG, GO, and KEGG databases, respectively. Additionally, 501 novel genes (20,326 novel transcripts) were identified for the first time, among which 401 (20,255), 193 (13,365), 414 (19,186), 228 (12,093), and 202 (11,703) were respectively annotated to each of the aforementioned five databases. The expression and sequences of three randomly selected novel transcripts were confirmed by RT-PCR and Sanger sequencing. The 5' UTR of 2082 genes, the 3' UTR of 2029 genes, and both the 5' and 3' UTRs of 730 genes were extended. Moreover, 17,345 SSRs, 14,789 complete ORFs, 1224 long non-coding RNAs (lncRNAs), and 650 transcription factors (TFs) from 37 families were detected. Findings from this work not only refine the annotation of the A. mellifera reference genome, but also provide a valuable resource and basis for relevant molecular and -omics studies.

Ontogeny of immunity and natural viral infection in Apis mellifera drones and workers

The most common viral diseases affecting honey bees (Apis mellifera) in Israel include deformed wing viruses (DWV-A and DWV-B) and acute paralysis viruses (ABPV and IAPV). These viruses are transmitted within and between colonies, both horizontally and vertically. All members of the colony contribute to this transmission, on the other hand individual and social immunity, particularly hygienic behaviour, may affect the outcome of the process. In this study, we evaluated the ontogeny of natural infections of DWV-A, DWV-B, ABPV and IAPV, their prevalence and loads, in workers and drones from high (H) and low (L) hygienic colonies. In parallel, we evaluated the expression of two immune genes: peptidoglycan recognition protein S2(PGRP-S2) and hymenoptaecin. The prevalence of DWV-B and IAPV increased with age and was higher in workers than in drones. ABPV was not detected in drones. The expression of both immune genes was significantly affected by age and sex. Drones from H colonies had higher expression of these genes. The increased expression of immune genes with drones’ age, particularly in hygienic colonies, suggest additional value of honey bee breeding for hygienic behaviour for sustainable beekeeping.

Expression and function of the vitellogenin receptor in the hypopharyngeal glands of the honey bee Apis mellifera (Hymenoptera: Apidae) workers.

The vitellogenin receptor (VgR) is essential for the uptake and transport of the yolk precursor, vitellogenin (Vg). Vg is synthesized in the fat body, released in the hemolymph, and absorbed in the ovaries, via receptor-mediated endocytosis. Besides its important role in the reproductive pathway, Vg occurs in nonreproductive worker honey bee, suggesting its participation in other pathways. The objective was to verify if the VgR occurs in the hypopharyngeal glands of Apis mellifera workers and how Vg is internalized by these cells. VgR occurrence in the hypopharyngeal glands was evaluated by qPCR analyses of VgR and immunohistochemistry in workers with different tasks. The VgR gene is expressed in the hypopharyngeal glands of workers with higher transcript levels in nurse honey bees. VgR is more expressed in 11-day-old workers from queenright colonies, compared to orphan ones. Nurse workers with developed hypopharyngeal glands present higher VgR transcripts than those with poorly developed glands. The immunohistochemistry results showed the co-localization of Vg, VgR and clathrin (protein that plays a major role in the formation of coated vesicles in endocytosis) in the hypopharyngeal glands, suggesting receptor-mediated endocytosis. The results demonstrate that VgR performs the transport of Vg to the hypopharyngeal glands, supporting the Ovary Ground Plan Hypothesis and contributing to the understanding of the role of this gland in the social context of honey bees.

Exploring the effects of the acaricide cyflumetofen on the vital organs of the honey bee Apis mellifera (Hymenoptera: Apidae) workers

Bees are important for maintaining ecosystems, pollinating crops and producing marketable products. In recent years, a decline in bee populations has been reported, with multifactorial causes, including the intensification of pesticide use in agriculture. Among pesticides, cyflumetofen is an insecticide and acaricide used in apple, coffee and citrus crops, whose main pollinator is the honey bee Apis mellifera. Therefore, this bee is a potential target of cyflumetofen during foraging. This study evaluated the histopathological and cytological damage in the midgut, hypopharyngeal glands and fat body of A. mellifera workers exposed to LC50 of cyflumetofen. The midgut epithelium of exposed bees presented cytoplasmic vacuolization, release of vesicles and cell fragments, which indicate autophagy, increased production of digestive enzymes and cell death, respectively. The cytological analysis of the midgut revealed the dilation of the basal labyrinth and the presence of spherocrystals in the digestive cells. The hypopharyngeal glands produced greater amounts of secretion in treated bees, whereas no changes were observed in the fat body. The results indicate that acute exposure to cyflumetofen negatively affect A. mellifera, causing damage to the midgut and changes in the hypopharyngeal glands, which may compromise the survival and foraging of this pollinator.