Translocation of Tebuconazole between Bee Matrices and Its Potential Threat on Honey Bee (Apis mellifera Linnaeus) Queens

Various pesticide residues can be found in different bee colony components. The queen larvae of honey bee (Apis mellifera L.) receive non-contaminated food from nurse bees. However, there is little knowledge about how pesticide residues affect developing bees. Additionally, little is known about the migration of lipophilic pesticides between bee matrices. While wax, royal jelly (RJ), and bee larvae are chemically distinct, they all contain lipids and we expected the lipophilic fungicide tebuconazole to be absorbed by different contacting materials. Our aim was to analyze the translocation of tebuconazole residues from queen cell wax to RJ, queen larvae, and newly emerged queens and to evaluate its potential risk to queens. We demonstrated the potential for the migration of tebuconazole from wax to RJ, with a strong dilution effect from the original contamination source. No residues were detected in queen bee larvae and newly emerged queens, indicating that the migration of tebuconazole probably did not directly endanger the queen bee, but there was some risk that tebuconazole might still affect the homeostasis of developing bees.

Genetic diversity among queen bee, worker bees and larvae in terms of retrotransposon movements

Objective Honeybee (Apis mellifera L.) is a model organism, contributing significant effect on global ecology by pollination and examining due to its social behaviour. Methods In this study, barley-specific Sukkula and Nikita retrotransposons were analysed using IRAP (Inter-Retrotransposon Amplification Polymorphism) marker technique, and the relationships between retrotransposon movements and development were also investigated in three different colonies of the Caucasian bee (Apis mellifera caucasica). Furthermore, transposon sequences belonging to Apis mellifera, Bombus terrestris, Triticum turgidum and Hordeum vulgare were also examined to figure out evolutionary relationships. Results For this purpose, a queen bee, five worker bees, and five larvae from each colony were studied. Both retrotransposons were found in all samples in three colonies with different polymorphism ratios (0-100% for Nikita and 0-67% for Sukkula). We also determined polymorphisms in queen-worker (0-83% for Nikita, 0-63% for Sukkula), queen-larvae (0-83% for Nikita, 0-43% for Sukkula) and worker-larvae comparisons (0-100% for Nikita, 0-63% for Sukkula) in colonies. Moreover, close relationships among transposons found in plant and insect genomes as a result of in silico evaluations to verify experimental results. Conclusion This work could be one of the first studies to analyse plant-specific retrotransposons’ movements in honeybee genome. Results are expected to understand evolutionary relationships in terms of horizontal transfer of transposons among kingdoms.

Morphology of immatures of the thelytokous ant, Monomorium triviale Wheeler (Formicidae: Myrmicinae: Solenopsidini) with descriptions of the larval caste dimorphism

The ant genus Monomorium is one of the most species-rich but taxonomically problematic groups in the hyperdiverse subfamily Myrmicinae. An East Asian species, M. triviale Wheeler, W. M., produces both reproductive queens and sterile workers via obligate thelytokous parthenogenesis. Here, we describe the immature forms of M. triviale based on light and scanning electron microscopy observations, with a note on the striking caste dimorphism in the last larval instar. The last-instar queen larvae were easily recognized by their large size, "aphaenogastoroid" body shape, and rows of doorknob-like tubercles on the lateral and dorsal body surface. This type of queen-specific structure has not been found in ants in general, let alone congeneric species found in Japan. In stark contrast to the queen larvae, worker larvae showed a "pheidoroid" body shape and a body surface similar to other ants. The worker larvae were estimated to have three instars, consistent with previously described congeners. The pupae of both castes also had no cocoon, a characteristic commonly described in other Myrmicinae species. In total, the developmental period from egg to adult worker averaged 59 days under 25℃. We discuss possible functions of the tubercles of queen larvae based on previous studies.

Pesticide Exposure During Development Does Not Affect the Larval Pheromones, Feeding Rates, or Morphology of Adult Honey Bee (Apis mellifera) Queens

Recent work demonstrated that honey bee (Apis mellifera L.) queens reared in pesticide-laden beeswax exhibit significant changes in the composition of the chemicals produced by their mandibular glands including those that comprise queen mandibular pheromone, which is a critical signal used in mating as well as queen tending behavior. For the present study, we hypothesized that pesticide exposure during development would alter other queen-produced chemicals, including brood pheromone in immature queens, thus resulting in differential feeding of queen larvae by nurse workers, ultimately impacting adult queen morphology. We tested these hypotheses by rearing queens in beeswax containing field-relevant concentrations of (1) a combination of tau-fluvalinate and coumaphos, (2) amitraz, or (3) a combination of chlorothalonil and chlorpyrifos. These pesticides are ubiquitous in most commercial beekeeping operations in North America. We observed nurse feeding rates of queen larvae grafted into pesticide-laden beeswax, analyzed the chemical composition of larval queen pheromones and measured morphological markers in adult queens. Neither the nurse feeding rates, nor the chemical profiles of immature queen pheromones, differed significantly between queens reared in pesticide-laden wax compared to queens reared in pesticide-free wax. Moreover, pesticide exposure during development did not cause virgin or mated adult queens to exhibit differences in morphological markers (i.e., body weight, head width, or thorax width). These results were unexpected given our previous research and indicate that future work is needed to fully understand how pesticide exposure during development affects honey bee queen physiology, as well as how various adult queen quality metrics relate to each other.

Experimental increase in fecundity causes upregulation of fecundity and body maintenance genes in the fat body of ant queens

In most organisms, fecundity and longevity are negatively associated and the molecular regulation of these two life-history traits is highly interconnected. In addition, nutrient intake often has opposing effects on lifespan and reproduction. In contrast to solitary insects, the main reproductive individual of social hymenopterans, the queen, is also the most long-lived. During development, queen larvae are well-nourished, but we are only beginning to understand the impact of nutrition on the queens' adult life and the molecular regulation and connectivity of fecundity and longevity. Here, we used two experimental manipulations to alter queen fecundity in the ant Temnothorax rugatulus and investigated associated changes in fat body gene expression. Egg removal triggered a fecundity increase, leading to expression changes in genes with functions in fecundity such as oogenesis and body maintenance. Dietary restriction lowered the egg production of queens and altered the expression of genes linked to autophagy, Toll signalling, cellular homeostasis and immunity. Our study reveals that an experimental increase in fecundity causes the co-activation of reproduction and body maintenance mechanisms, shedding light on the molecular regulation of the link between longevity and fecundity in social insects.

Prisoners receive food fit for a queen: honeybees feed small hive beetles protein-rich glandular secretions through trophallaxis

The honeybee nest parasite Aethina tumida (small hive beetle), uses behavioural mimicry to induce trophallactic feeding from its honeybee hosts. Small hive beetles are able to induce honeybee workers to share the carbohydrate–rich contents of their crops, but it is not clear whether the beetles are able to induce to workers to feed them the protein-rich hypopharyngeal glandular secretions fed to the queen, larvae and other nest mates. Protein is a limiting macronutrient in an insect's diet, essential for survival, growth and fecundity. Honeybees obtain protein from pollen, which is consumed and digested by nurse bees. They then distribute the protein to the rest of the colony in the form of hypopharyngeal gland secretions. Using 14C-phenylalanine as a qualitative marker for protein transfer, we show that small hive beetles successfully induce worker bees to feed them the protein-rich secretions of their hypopharyngeal glands during trophallaxis, and that females are more successful than males in inducing the transfer of these protein-rich secretions. Furthermore, behavioural observations demonstrated that female beetles do not preferentially interact with a specific age cohort of bees when soliciting food, but males tend to be more discriminate and avoids the more aggressive and active older bees.

Virion Structure of Black Queen Cell Virus, a Common Honeybee Pathogen

ABSTRACTViral diseases are a major threat to honeybee (Apis mellifera) populations worldwide and therefore an important factor in reliable crop pollination and food security. Black queen cell virus (BQCV) is the etiological agent of a fatal disease of honeybee queen larvae and pupae. The virus belongs to the genusTriatovirusfrom the familyDicistroviridae, which is part of the orderPicornavirales. Here we present a crystal structure of BQCV determined to a resolution of 3.4 Å. The virion is formed by 60 copies of each of the major capsid proteins VP1, VP2, and VP3; however, there is no density corresponding to a 75-residue-long minor capsid protein VP4 encoded by the BQCV genome. We show that the VP4 subunits are present in the crystallized virions that are infectious. This aspect of the BQCV virion is similar to that of the previously characterized triatoma virus and supports the recent establishment of the separate genusTriatoviruswithin the familyDicistroviridae. The C terminus of VP1 and CD loops of capsid proteins VP1 and VP3 of BQCV form 34-Å-tall finger-like protrusions at the virion surface. The protrusions are larger than those of related dicistroviruses.IMPORTANCEThe western honeybee is the most important pollinator of all, and it is required to sustain the agricultural production and biodiversity of wild flowering plants. However, honeybee populations worldwide are suffering from virus infections that cause colony losses. One of the most common, and least known, honeybee pathogens is black queen cell virus (BQCV), which at high titers causes queen larvae and pupae to turn black and die. Here we present the three-dimensional virion structure of BQCV, determined by X-ray crystallography. The structure of BQCV reveals large protrusions on the virion surface. Capsid protein VP1 of BQCV does not contain a hydrophobic pocket. Therefore, the BQCV virion structure provides evidence that capsid-binding antiviral compounds that can prevent the replication of vertebrate picornaviruses may be ineffective against honeybee virus infections.

Rearing Drones in Queen Cells of Apis mellifera Honey Bees

We investigated the rearing of drone larvae grafted in queen cells. From the 1200 drone larvae that were grafted during spring and autumn, 875 were accepted (72.9%) and reared as queens. Drone larvae in false queen cells received royal jelly of the same composition and of the same amounts as queen larvae. Workers capped the queen cells as if they were drones, 9-10 days after the egg laying. Out of 60 accepted false queen cells, 21 (35%) were capped. The shape of false queen cells with drone larvae is unusually long with a characteristically elongate tip which is probably due to the falling of larvae. Bees start the destruction of the cells when the larvae were 3 days old and maximised it before and after capping. Protecting false queen cells in the colony by wrapping, reversing them upside down, or placing in a horizontal position, did not help. The only adult drones that emerged from the false queen cells were those protected in an incubator and in push-in cages. Adult drones from false queen cells had smaller wings, legs, and proboscis than regular drones. The results of this study verify previous reports that the bees do not recognise the different sex of the larvae at least at the early stage of larval development. The late destruction of false queen cells, the similarity in quality and quantity of the produced royal jelly, and the bigger drone cells, allow for the use of drone larvae in cups for the production of royal jelly.