Gut microbiota composition is associated with environmental landscape in honey bees

There is growing recognition that the gut microbial community regulates a wide variety of important functions in its animal hosts, including host health. However, the complex interactions between gut microbes and environment are still unclear. Honey bees are ecologically and economically important pollinators that host a core gut microbial community that is thought to be constant across populations. Here, we examined whether the composition of the gut microbial community of honey bees is affected by the environmental landscape the bees are exposed to. We placed honey bee colonies reared under identical conditions in two main landscape types for 6 weeks: either oilseed rape farmland or agricultural farmland distant to fields of flowering oilseed rape. The gut bacterial communities of adult bees from the colonies were then characterized and compared based on amplicon sequencing of the 16S rRNA gene. While previous studies have delineated a characteristic core set of bacteria inhabiting the honey bee gut, our results suggest that the broad environment that bees are exposed to has some influence on the relative abundance of some members of that microbial community. This includes known dominant taxa thought to have functions in nutrition and health. Our results provide evidence for an influence of landscape exposure on honey bee microbial community and highlight the potential effect of exposure to different environmental parameters, such as forage type and neonicotinoid pesticides, on key honey bee gut bacteria. This work emphasizes the complexity of the relationship between the host, its gut bacteria, and the environment and identifies target microbial taxa for functional analyses.     

Social immunity in honeybees—Density dependence,diet, and body mass trade‐offs

Group living is favorable to pathogen spread due to the increased risk of disease transmission among individuals. Similar to individual immune defenses, social immunity, that is antiparasite defenses mounted for the benefit of individuals other than the actor, is predicted to be altered in social groups. The eusocial honey bee (Apis mellifera) secretes glucose oxidase (GOX), an antiseptic enzyme, throughout its colony, thereby providing immune protection to other individuals in the hive. We conducted a laboratory experiment to investigate the effects of group density on social immunity, specifically GOX activity, body mass and feeding behavior in caged honey bees. Individual honeybees caged in a low group density displayed increased GOX activity relative to those kept at a high group density. In addition, we provided evidence for a trade‐off between GOX activity and body mass: Individuals caged in the low group density had a lower body mass, despite consuming more food overall. Our results provide the first experimental evidence that group density affects a social immune response in a eusocial insect. Moreover, we showed that the previously reported trade‐off between immunity and body mass extends to social immunity. GOX production appears to be costly for individuals, and potentially the colony, given that low body mass is correlated with small foraging ranges in bees. At high group densities, individuals can invest less in social immunity than at low densities, while presumably gaining shared protection from infection. Thus, there is evidence that trade‐offs at the individual level (GOX vs. body mass) can affect colony‐level fitness.     

Floral volatile sesquiterpenes of Elsholtzia rugulosa (Lamiaceae) selectively attract Asian honey bees

There are few floral volatiles of compounds that have been properly assessed for attracting pollinators. The intense plant odour of Elsholtzia rugulosa (Lamiaceae) to humans may be attributed to the presence of high concentrations of β‐caryophyllene. In a previous study, Zhang, Yang, and Zhang (Scientific Reports, 6, 2016a, 276161) speculated that the presence of β‐caryophyllene attracts pollinators (e.g., honey bees) to its flowers, an assumption that was assessed through the exploration of the functional significance of specific floral volatile compounds and the evaluation of their effects on the behaviour of Apis cerana (Asian honey bees; a known pollinator) and two non‐pollinators, a hornet (Vespa velutina) and a bumblebee (Bombus sp.). The results from these behavioural experiments indicated that both β‐caryophyllene and β‐elemene (both naturally found in flowers of E. rugulosa) were individually attractive to the Asian honey bees. Moreover, the combination of the two was more effective than either of them separately. In contrast, neither compound, nor a blend of the two at different concentrations, was attractive to the hornet and bumblebee species. These results demonstrate that β‐caryophyllene and β‐elemene play a key signalling role in attracting Asian honey bees to E. rugulosa.     

The biosynthesis of Dufour's gland constituents in queens of the honeybee (Apis mellifera)

Chemical analysis of queen Dufour's gland by GC/MS revealed the presence of long chain esters and hydrocarbons. To evaluate whether the biosynthesis of Dufour's gland components is modulated by physiological factors, we studied thede novo biosynthesis of esters and hydrocarbons in virgin and mated queen glandsin vivo andin vitro. Using [1-¹⁴C] sodium acetate as a precursor, it was shown thatin vivo, esters and hydrocarbons comprised the majorde novo products. The relative proportion of the hydrocarbon fraction was significantly higher in mated queens than in virgins, while in esters the situation was reversed. Comparision between reproductive and non reproductive virgin queens revealed that mating and not egg laying induced changes in glandular activity. Glands incubatedin vitro, on the other hand, synthesized mainly esters and alcohols but not hydrocarbons. These results suggest that either hydrocarbons are produced elsewhere and subsequently sequestered by Dufour's gland, or that the gland biosynthetic expressionin vivo is modulated. Differences between the gland activityin vivo andin vitro further support the hypothesis of a physiological mechanism in regulation of glandular performance.     

CLONING AND SEQUENCING OF GENE ENCODING HYALURONIDASE FROM THE VENOM OF APIS CERANA CERANA*

Abstract The venomous hyaluronidase (Hya) gene of Chinese honey bee, Apis cerana cerana, was amplified by RT‐PCR from total RNA of venom glands of the worker bees. The full length of its nucleotide is 1164 bp encoding a 387 amino acid polypeptide with predicted molecular weight of 42.6 kD. The alignment of AcHya amino acid sequence with other 6 Hyas shows that AcHya is most closely related to the Hya of European honey bee, A. mellifera, with 91% amino acid identity. It also shares homology with Hya of Dolichovespul amacidata, Polistes annularis, Vespula vulgaris, Lutzomia longipalpis and Homo sapiens (sperm), with 54%., 52%., 46%, 27% and 20% amino acid identity, respectively. A phylogenetic tree of those hyaluronidases was drawn by using GENETYX program, the conservation, the relationship between molecular structure and function of 7 hyaluronidases as above was compared and analysed.     

Lipid-enhanced pollen and lipid-reduced flour diets and their effect on the longevity of honey bees (Apis mellifera L.)

Abstract  As eucalypt pollens contain low concentrations of lipid, enhancing pollen diets with fatty acids was hypothesised to improve honey bee longevity. Different concentrations of linoleic and oleic acid added to eucalypt pollen were trialled in small cages containing approximately 1400 bees each. Bees fed diets of redgum ( Corymbia calophylla (Lindl.) Hill & Johnson, formerly Eucalyptus calophylla ) pollen had the lowest mortality of 22 diets tested for 6 weeks and had life spans (50%) greater than 42 days. Linoleic acid mixed with a redgum diet in concentrations >6% corresponded to life spans of 24–25 days. Bee longevity appeared to be more sensitive to oleic acid as life spans decreased to 15–21 days when diets had concentrations >2%. The life spans of bees fed soya bean flour were 26 days on low (0.6% lipid) fat, 19 days on defatted and 20 days on full-fat diets. Bees fed lupin flour had a life span of 23 days. Adding redgum pollen to lupin flour caused increased mortality, but addition of pollen to soya bean flour was beneficial. Thus, beekeepers who choose to utilise soya bean or lupin flours as protein substitutes to pollen will have bees with reduced longevity. Bees fed redgum pollen that had been dried, crushed, irradiated and hermetically stored in a cool room for several years had similar longevity to bees fed fresh-collected and frozen redgum pollen.     

Brain melatonin content and polyethism in adult workers of Apis mellifera and Apis cerana (Hym., Apidae)

Abstract:  Radioimmunoassay showed that melatonin, N -acetyl-5-methoxytryptamine, was synthesized in the brain of workers of Apis mellifera ligustica Spinola, and related to the polyethism of the workers. Statistically significant differences were found in melatonin levels between workers at different ages and between workers performing different tasks. The level in foragers was higher than that in young nurses. The highest melatonin level was found in 21-day-old adult workers (213.15 pg per worker head) and the lowest level in 1-day-old adult workers (6.08 pg per worker head); and the highest level in pollen-collecting workers (404.13 pg per worker head) and the lowest in nurse workers (13.10 pg per worker head). Apis cerana cerana Fabricius showed a similar pattern, but the nectar foragers had the highest melatonin levels. The melatonin level also showed a daily rhythm. In 30-day-old adult workers there were two peaks, at 7:00 and 19:00 hours, and three troughs at 15:00, 21:00 and 1:00 hours respectively.     

The effect of genotype on response thresholds to sucrose and foraging behavior of honey bees (Apis mellifera L.)

Honey bee foragers were tested for their proboscis extension response (PER) to water and varying solutions of sucrose. Returning pollen and nectar foragers were collected at the entrance of a colony and were assayed in the laboratory. Pollen foragers had a significantly higher probability of responding to water and to lower concentrations of sucrose. Bees derived from artificially selected high- and low-pollen-hoarding strains were also tested using the proboscis extension assay. Returning foragers were captured and tested for PERs to 30% sucrose. Results demonstrated a genotypic effect on PERs of returning foragers. The PERs of departing high- and low-strain foragers were consistent with those of returning foragers. The PERs were related to nectar and water reward perception of foragers. High strain bees were more likely to return with loads of water and lower concentrations of sucrose than foragers from the low pollen strain. Low-strain bees were more likely to return empty. We identified a previously mapped genomic region that contains a variable quantitative trait locus that appears to influence sucrose response thresholds. These studies demonstrate a gene-brain-behavior pathway that can be altered as a consequence of colony-level selection for quantities of stored food. Accepted: 3 September 1997     

Floral resources and risk of exposure to pesticides for Melipona quadrifasciata anthidioides Lepeletier 1836 in a Cerrado of São Paulo (Brazil)

Honey and bee bread samples from storage pots of Melipona quadrifasciata anthidioides were collected monthly from April 2015 to May 2016 in the Mogi Guaçu Biological Reserve (22º 10? S, 47º 11? W). The flora in the site is characteristic of the Atlantic Forest with preserved areas of savanna-like vegetation surrounded by commercial forests, orchards and various crops of exotic and native plants. Samples were analysed with the use of melissopalynological methodology and 46 pollen types from 38 genera and 30 families were identified in 25 honey samples. Fabaceae, Asteraceae, Myrtaceae, Sapindaceae showed the greatest pollen richness in honey. Predominant nectariferous pollen types were Anadenanthera, Cordia, Eucalyptus, Mimosa scabrella, Schefflera, Sida, Serjania and Vernonia. Twenty-eight types of pollen from 21 genera and 19 families were identified in 22 bee bread samples. Fabaceae, Asteraceae and Myrtaceae showed the highest pollen richness. Anadenanthera, Cecropia, Eucalyptus, Melastomataceae, Mimosa scabrella, Mimosa verrucosa and Myrcia were the most frequent polliniferous pollen types. Principal component analysis (PCA) demonstrated that honey and pollen samples formed two main groups of similarity, mainly due to Eucalyptus’ nectar and pollen of Melastomataceae, respectively. Melipona quadrifasciata anthidioides collected nectar and pollen from the preserved areas as well as in the secondary and ‘ruderal’ vegetation and in cultivated forests/fields, suggesting their importance as pollinators both of native flora and exotic species. The use of trophic resources of plants grown with pesticides is a concern for the conservation of these species of bee and should be better studied.