Origin and Effect of Alpha 2.2 Acetobacteraceae in Honey Bee Larvae and Description of Parasaccharibacter apium gen. nov., sp. nov.

ABSTRACTThe honey bee hive environment contains a rich microbial community that differs according to niche.AcetobacteraceaeAlpha 2.2 (Alpha 2.2) bacteria are present in the food stores, the forager crop, and larvae but at negligible levels in the nurse and forager midgut and hindgut. We first sought to determine the source of Alpha 2.2 in young larvae by assaying the diversity of microbes in nurse crops, hypopharyngeal glands (HGs), and royal jelly (RJ). Amplicon-based pyrosequencing showed that Alpha 2.2 bacteria occupy each of these environments along with a variety of other bacteria, includingLactobacillus kunkeei. RJ and the crop contained fewer bacteria than the HGs, suggesting that these tissues are rather selective environments. Phylogenetic analyses showed that honey bee-derived Alpha 2.2 bacteria are specific to bees that “nurse” the hive's developing brood with HG secretions and are distinct from theSaccharibacter-type bacteria found in bees that provision their young differently, such as with a pollen ball coated in crop-derived contents.Acetobacteraceaecan form symbiotic relationships with insects, so we next tested whether Alpha 2.2 increased larval fitness. We cultured 44 Alpha 2.2 strains from young larvae that grouped into nine distinct clades. Three isolates from these nine clades flourished in royal jelly, and one isolate increased larval survivalin vitro. We conclude that Alpha 2.2 bacteria are not gut bacteria but are prolific in the crop-HG-RJ-larva niche, passed to the developing brood through nurse worker feeding behavior. We propose the nameParasaccharibacter apiumfor this bacterial symbiont of bees in the genusApis.

Routes of Acquisition of the Gut Microbiota of the Honey Bee Apis mellifera

ABSTRACTStudies of newly emergedApis melliferaworker bees have demonstrated that their guts are colonized by a consistent core microbiota within several days of eclosure. We conducted experiments aimed at illuminating the transmission routes and spatiotemporal colonization dynamics of this microbiota. Experimental groups of newly emerged workers were maintained in cup cages and exposed to different potential transmission sources. Colonization patterns were evaluated using quantitative real-time PCR (qPCR) to assess community sizes and using deep sequencing of 16S rRNA gene amplicons to assess community composition. In addition, we monitored the establishment of the ileum and rectum communities within workers sampled over time from natural hive conditions. The study verified that workers initially lack gut bacteria and gain large characteristic communities in the ileum and rectum within 4 to 6 days within hives. Typical communities, resembling those of workers within hives, were established in the presence of nurse workers or nurse worker fecal material, and atypical communities of noncore or highly skewed compositions were established when workers were exposed only to oral trophallaxis or hive components (comb, honey, bee bread). The core species of Gram-negative bacteria,Snodgrassella alvi,Gilliamella apicola, andFrischella perrara, were dependent on the presence of nurses or hindgut material, whereas some Gram-positive species were more often transferred through exposure to hive components. These results indicate aspects of the colony life cycle and behavior that are key to the propagation of the characteristic honey bee gut microbiota.

The effect of using prebiotic and probiotic products on intestinal micro-flora of the honeybee (Apis mellifera carpatica)

Maintaining bee colonies in a healthy state throughout the year is one of the main concerns of apiculture researchers. The phenomenon of disappearance of bee colonies is determined by several factors, one of which is bee disease. Due to the organizational structure of the bee colony, disease transmission is rapid, especially through infected food or via the nurse worker bees that feed the brood bees of the colony concerned. The practice of stimulating the bee colonies in spring using sugar syrup feeds with added prebiotic products (lactic acid or acetic acid) and probiotics (Lactobacillus acidophilus LA-14 and Bifidobacterium lactis BI-04) by using an Enterobiotic product (Lactobacillus casei), marketed as Enterolactis Plus, for three weeks, resulted in a significant reduction of the total number of bacteria in the digestive tracts of the bees, compared with the control group. By contrast, intestinal colonization with beneficial bacteria contained in probiotics products administered to the bees was observed. This resulted in an improved health status and bio productive index of the bee colonies studied.