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Honeybees are prone to poisoning after collecting jujube nectar during the jujube flowering period ('honeybee's jujube flower disease'). To explore the mechanism of honeybee poisoning, the gut microbiota of honeybees undergoing the disease were characterised based on amplicon sequencing of the 16 S rRNA gene. Our results showed that the composition and diversity of the gut microbiota were significantly altered in diseased honeybees. We observed a decrease in the relative abundance of Proteobacteria and increased abundances of Firmicutes and Actinobacteria in the midgut and hindgut of diseased honeybees. Moreover, linear discriminant analysis (LDA) effect size revealed significantly selected enrichment of Fructobacillus and Snodgrassella in the midguts from diseased honeybees and Lactobacillus, Bifidobacterium, and Snodgrassella in the hindguts from diseased honeybees. Tax4Fun anylasis indicated that the functional potential of the diseased honeybee gut bacterial community was significantly changed relative to the healthy honeybee. Carbohydrate metabolism, nucleotides metabolism, amino acid synthesis metabolism, coenzyme and vitamins metabolism were increased, while energy metabolism and xenobiotic biodegradation and metabolism were decreased in the diseased honeybees. These results provide a new perspective for evaluating the response of honeybees to jujube flower disease based on changes in the intestinal microflora., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier Inc.)

Temperature and humidity are the most important factors affecting the growth, reproduction, and survival of bees. Apis mellifera are important pollinating bees that are widely used in agricultural systems. However, the higher temperatures and humidity in greenhouses are not conducive to the survival of bees. Although previous research has revealed the behavioral responses and physiological mechanisms of honeybees to adapt to high temperature and humidity, there are few data on the exact molecular mechanisms involved. In our study, we investigated gene expression in A. mellifera under different temperature and humidity treatments, using transcriptomic analysis to identify differentially expressed genes (DEGs) and relevant biological processes. Based on the transcriptomic results, we selected several genes with significant differences in expression, and detected the expression patterns of these genes at different temperatures or humidity or different treatment times by q-RT PCR. In the high temperature treatments, 434 DEGs were identified; in the high humidity treatments, 86 DEGs were identified; in the combined high temperature and humidity treatments, 266 DEGs were identified. Analysis results showed that DEGs were enriched in pathways related to amino acid and fatty acid biosynthesis and metabolism under each treatment. In addition, heat shock proteins, zinc finger proteins, serine/threonine-protein kinases, and antioxidase were differentially expressed between the different treatments. The results of the q-RT PCR showed that the expression levels of these genes increased with increasing temperature and over treatment time. Our findings provide a general expression profile of the adaptive expression of heat-resistance genes responding to high temperature and high humidity in A. mellifera, including the expression patterns of several DEGs. Our data provide a basis for future research on the mechanisms underlying the adaptation of insects to high temperature and humidity., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The ambient temperature and relative humidity affect the metabolic and physiological responses of bees, thus affecting their life activities. However, the physiological changes in bee due to high temperature and high humidity remain poorly understood. In this study, we explored the effects of higher temperature and humidity on the epiphysiology of bees by evaluating the survival, tolerance and body water loss in two bee species (Apis cerana and Apis mellifera). We also evaluated the changes in the activity of antioxidant and detoxification enzymes in their body. We observed that under higher temperature and humidity conditions, the survival rate of A. mellifera was higher than that of A. cerana. On the other hand, a comparison of water loss between the two species revealed that A. mellifera lost more water. However, under extremely high temperature conditions, A. cerana was more tolerant than A. mellifera. Moreover, under higher temperature and humidity conditions, the activity of antioxidant and detoxification enzymes in bees was significantly increased. Overall, these results suggest that high temperatures can adversely affect bees. They not only affect the survival and water loss, but also stimulate oxidative stress in bees. However, unlike our previous understanding, high humidity can also adversely affect bees, although its effects are lower than that of temperature., Competing Interests: The authors have declared that no competing interests exist.

Apis cerana cerana, an important endemic honey bee species in China, possesses valuable characteristics such as a sensitive olfactory system, good foraging ability, and strong resistance to parasitic mites. Here, we performed transcriptome sequencing of the antenna, the major chemosensory organ of the bee, using an Illumina sequencer, to identify typical differentially expressed genes (DEGs) in adult worker bees of different ages, namely, T1 (1 day); T2 (10 days); T3 (15 days); and T4 (25 days). Surprisingly, the expression levels of DEGs changed significantly between the T1 period and the other three periods. All the DEGs were classified into 26 expression profiles by trend analysis. Selected trend clusters were analyzed, and valuable information on gene expression patterns was obtained. We found that the expression levels of genes encoding cuticle proteins declined after eclosion, while those of immunity-related genes increased. In addition, genes encoding venom proteins and major royal jelly proteins were enriched at the T2 stage; small heat shock proteins showed significantly higher expression at the T3 stage; and some metabolism-related genes were more highly expressed at the T4 stage. The DEGs identified in this study may serve as a valuable resource for the characterization of expression patterns of antennal genes in A. cerana cerana. Furthermore, this study provides insights into the relationship between labor division in social bees and gene function., (Copyright © 2018. Published by Elsevier B.V.)

In social insects, the foraging gene (for) regulates insect age- and task-based foraging behaviors. We studied the expression and localization of the for gene (Acfor) in Apis cerana cerana workers to explore whether the differential regulation of this gene is associated with the behaviors of nurses and foragers. The expression profiles of Acfor in different tissues and at different ages were examined using real-time quantitative reverse transcription polymerase chain reaction. Cellular localization in the brain was detected using in situ hybridization. Acfor transcripts in different ages workers showed that Acfor expression was detected in all the heads of 1- to 30-d-old worker bees. Acfor expression reached a peak at 25 d of age, and then declined with increasing age. The results showed that Acfor gene expression in five tissues was respectively significantly higher in foragers than in nurses. In nurses, the relative expression of Acfor was the highest in the antennae. There was a highly significant difference in expression between antennae, legs, and the other three tissues. In foragers, Acfor expression was the highest in the thorax, which was significantly different from all other tissues. In situ hybridization showed that Acfor was highly expressed in the lamina of the optic lobes, in a central column of Kenyon cells in the mushroom bodies of the brain of workers, and in the antennal lobes. This suggested that Acfor expression affects age-related foraging behavior in Apis cerana cerana, and that it may be related to flight activity.

One important concern regarding the use of transgenic cotton expressing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) is its potential detrimental effect on non-target organisms. The honey bee (Apis mellifera) is the most important pollinator species worldwide and it is directly exposed to transgenic crops by the consumption of genetically modified (GM) pollen. However, the potential effects of Bt cotton on A. mellifera remain unclear. In the present study, we assessed the effects of two Bt cotton varieties; ZMSJ expressing the Cry1Ac and Cry2Ab insecticidal proteins, and ZMKCKC producing Cry1Ac and EPSPS, on A. mellifera. Feeding on pollen from two Bt cotton varieties led to detection of low levels of Cry toxins (<10 ng/g fresh weight) in the midgut of A. mellifera adults, yet expression of detoxification genes did not change significantly compared to feeding on non-Bt cotton. Binding assays showed no Cry1Ac or Cry2Ab binding to midgut brush border membrane proteins from A. mellifera adults. Taken together, these results support minimal risk for potential negative effects on A. mellifera by exposure to Bt cotton., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Insects use olfaction to recognize a wide range of volatile cues, to locate food sources, mates, hosts and oviposition sites. These chemical volatiles are perceived by odorant receptors (ORs) expressed on the dendritic membrane of olfactory neurons, most of which are housed within the chemosensilla of antennae. Most insect ORs are tandemly arrayed on chromosomes and some of them are formed by gene duplication. Here, we identified a pair of duplicated Or genes, AcerOr1 and AcerOr3, from the antennae of the Asian honeybee, Apis cerana cerana, and reported their molecular characterization and temporal expression profiles. The results showed that these two genes shared high similarity both in sequence and the gene structure. Quantitative real-time PCR analysis of temporal expression pattern indicated that in drones the expression pattern of these two genes were very similar. The transcripts expressed weakly in larvae and pupae, then increased gradually in adults. In workers, the expression level of AcerOr1 changed more drastically and expressed higher than that of AcerOr3. However, both reached their highest expression level in one-day-old adults. In addition, the expression profiles between different sexes revealed that AcerOr3 appear to be expressed biased in male antennae. These results suggest that AcerOr1 may perceive odours of floral scents, while AcerOr3 may detect odours critical to male behaviour, such as the queen substance cues.

Transgenic insect-resistant cotton (Bt cotton) has been extensively planted in China, but its effects on non-targeted insect species such as the economically important honey bee (Apis mellifera) and silkworm (Bombyx mori) currently are unknown. In this study, pollen from two Bt cotton cultivars, one expressing Cry1Ac/EPSPS and the other expressing Cry1Ac/Cry2Ab, were used to evaluate the effects of Bt cotton on adult honey bees and silkworm larvae. Laboratory feeding studies showed no adverse effects on the survival, cumulative consumption, and total hemocyte count (THC) of A. mellifera fed with Bt pollen for 7 days. No effects on the survival or development of B. mori larvae were observed either. A marginally significant difference between Cry1Ac/Cry2Ab cotton and the conventional cotton on the THC of the 3(rd) day of 5(th) B. mori instar larvae was observed only at the two highest pollen densities (approximately 900 and 8000 grains/cm(2)), which are much higher than the pollen deposition that occurs under normal field conditions. The results of this study show that pollen of the tested Bt cotton varieties carried no lethal or sublethal risk for A. mellifera, and the risk for B. mori was negligible.

Olfaction recognition process is extraordinarily complex in insects, and the olfactory receptors play an important function in the process. In this paper, a highly conserved olfactory co-receptor gene, AcerOr2 (ortholog to the Drosophila melanogaster Or83b), cloned from the antennae of the Asian honeybee, Apis cerana cerana Fabricius (Hymenoptera: Apidae), using reverse transcriptase PCR and rapid amplification of cDNA ends. The full-length sequence of the gene was 1763 bp long, and the cDNA open reading frame encoded 478 amino acid residues, including 7 putative transmembrane domains. Alignment analysis revealed that AcerOr2 shares high homology (> 74%) with similar olfactory receptors found in other Hymenoptera species. The amino acid identity with the closely related species Apis mellifera reached 99.8%. The developmental expression analysis using quantitative real-time reverse transcriptase PCR suggested that the AcerOr2 transcript was expressed at a relatively low level in the larval stage, whereas it was expressed broadly in the pupal and adult stages, with a significantly high level on the days just before and after eclosion. In situ hybridization showed that AcerOr2 mRNA was expressed in sensilla placodea and on the basal region of the worker antennal cuticle, in accordance with the previous conclusions that the conserved genes are expressed in most olfactory receptor neurons.

Abnormal temperature will affect the normal physiological activities of bees. Similarly, heat stress can also affect the immune system of bees. However, the changes that occur in immune system in bees that are exposed to heat stress remain unclear. We want to understand the impact of heat stress on the immune system of social insects (Apis cerana and Apis mellifera), so we analyzed the expression patterns of three antimicrobial peptide genes (Defensin, Abaecin, Hymenoptaecin) at different temperatures and different treatment times. Our results provide information on the expression patterns of Defensin, Abaecin and Hymenoptaecin at high temperatures. The expression of these genes was found to be increasingly affected by an increase in temperature and treatment time. Heat stress seems to have activated immunity and this early activity may prevent infection under stressful conditions. Our findings provide a basis for determining the mechanisms by which bees immune system adapt to high temperatures. [ABSTRACT FROM AUTHOR]

The malvolio (mvl) gene plays an important role in the transition from nursing to foraging in honeybees (Apis mellifera). Apis cerana cerana (A. c. cerana) is a subspecies of the eastern honeybee, well-known for its pollinator role throughout China. Although A. c. cerana shares many characteristics with A. mellifera, it is unclear whether Acmvl plays a similar role to Ammvl in foraging behavior. In this study, Acmvl expression was quantified during the transition from nursing to foraging in A. c. cerana. Acmvl protein production was also characterized in different tissues in bees from three behavioral groups. Finally, in situ hybridization was used to describe Acmvl expression patterns in forager bee brains. Acmvl expression was low early in life but then showed a major peak, which suggests its role in labor division. Examination of tissue type revealed that Acmvl expression was highest in the thoraxes of nurse bees and the heads of forager bees. In bee brains, Acmvl was selectively expressed in the somata of Kenyon cells in the mushroom bodies, optic lobes and antennal lobes. Taken together, these findings suggest that Acmvl plays a role in the nurse–forager transition of A. c. cerana. [ABSTRACT FROM AUTHOR]