Bees play important roles in socio-economic development, biodiversity conservation, and ecosystem stability. However, during the cold season, resources become limited, leading to significant losses in bee colonies. Although many studies have described the characteristics of winter bees and demonstrated that notable changes occur in their gut microflora, the underlying mechanisms remain yet to be fully elucidated. Therefore, this study was conducted to compare the gut microbiota dynamics of overwintering bees. Sample acquisition involved randomly selecting ten colonies each from three bee farms containing Apis cerana (AC) and Apis mellifera (AM), followed by dissection for further analysis. DNA was extracted, and 16S rDNA sequencing, along with various bioinformatics tools, was used to assess microbial diversity, functional differences, and species comparisons between AC and AM gut microbiota. AC exhibited lower β diversity in the gut microbiota than AM during winter. Moreover, Gilliamella and Apibacter were relatively more abundant in AC. Regarding microbial functions, key pathways included the phosphotransferase system, galactose metabolism, the pentose phosphate pathway, and carbohydrate transport and metabolism. These results suggest the presence of microbial diversity differences between AC and AM, with the differential microbial functions mainly enriched in metabolic pathways that facilitate adaptation to cold environmental stress.

Honeybee gut microbiota plays a crucial role in maintaining their health and digestive function. Studies have confirmed that quercetin improves honeybee health by enhancing their pesticide tolerance and survival rates. This study aimed to examine the effects of quercetin on the bee gut microbiome by absolute quantification sequencing. We included 1800 bees from the experimental apiary and exposed them to 151.2, 75.6, and 37.8 mg/L of quercetin. Gut samples were collected on the 5th and 9th days, subjected to a polymerase chain reaction and 16S rRNA sequencing, and analyzed. After 5 days of quercetin treatment, the diversity of the honeybee gut microbiota was altered, and total bacterial copies and Lactobacillus abundance significantly decreased at high quercetin concentrations (151.2 and 75.6 mg/L). On day 9, the gut microbial community had recovered from the adverse effects, and Gilliamella abundance increased in response to 37.8 mg/L quercetin treatment. However, quercetin had no noticeable effects on survival rate, food consumption, and gut structure. Our study confirmed the effect of short-term quercetin intake on the gut microbiota of A. cerana, providing valuable insights into how phytochemicals alter the bee gut microbiome, and their repercussions on host physiology.