American foulbrood (AFB) disease and chalkbrood disease (CBD) are important bacterial and fungal diseases, respectively, that affect honeybee broods. Exposure to agrochemicals is an abiotic stressor that potentially weakens honeybee colonies. Gut microflora alterations in adult honeybees associated with these biotic and abiotic factors have been investigated. However, microbial compositions in AFB- and CBD-infected larvae and the profile of whole-body microbiota in foraging bees exposed to agrochemicals have not been fully studied. In this study, bacterial and fungal communities in healthy and diseased (AFB/CBD) honeybee larvae were characterized by amplicon sequencing of bacterial 16S rRNA gene and fungal internal transcribed spacer1 region, respectively. The bacterial and fungal communities in disordered foraging bees poisoned by agrochemicals were analysed. Our results revealed that healthy larvae were significantly enriched in bacterial genera Lactobacillus and Stenotrophomonas and the fungal genera Alternaria and Aspergillus. The enrichment of these microorganisms, which had antagonistic activities against the etiologic agents for AFB and CBD, respectively, may protect larvae from potential infection. In disordered foraging bees, the relative abundance of bacterial genus Gilliamella and fungal species Cystofilobasidium macerans were significantly reduced, which may compromise hosts' capacities in nutrient absorption and immune defence against pathogens. Significantly higher frequency of environmentally derived fungi was observed in disordered foraging bees, which reflected the perturbed microbiota communities of hosts. Results from PICRUSt and FUNGuild analyses revealed significant differences in gene clusters of bacterial communities and fungal function profiles. Overall, results of this study provide references for the composition and function of microbial communities in AFB- and CBD-infected honeybee larvae and foraging bees exposed to agrochemicals.

Chun-Xiang Yan

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