Host-derived organic acids enable gut colonization of the honey bee symbiont Snodgrassella alvi.

Diverse bacteria can colonize the animal gut using dietary nutrients or by engaging in microbial crossfeeding interactions. Less is known about the role of host-derived nutrients in enabling gut bacterial colonization. Here we examined metabolic interactions within the evolutionary ancient symbiosis between the honey bee (Apis mellifera) and the core gut microbiota member Snodgrassella alvi. This betaproteobacterium is incapable of metabolizing saccharides, yet colonizes the honey bee gut in the presence of a sugar-only diet. Using comparative metabolomics, 13C-tracers and nanoscale secondary ion mass spectrometry (NanoSIMS), we show in vivo that S. alvi grows on host-derived organic acids, including citrate, glycerate and 3-hydroxy-3-methylglutarate, which are actively secreted by the host into the gut lumen. S. alvi also modulates tryptophan metabolism in the gut by converting kynurenine to anthranilate. These results suggest that S. alvi is adapted to a specific metabolic niche in the honey bee gut that depends on host-derived nutritional resources.

Niche partitioning facilitates coexistence of closely related honey bee gut bacteria.

Ecological processes underlying bacterial coexistence in the gut are not well understood. Here, we disentangled the effect of the host and the diet on the coexistence of four closely related Lactobacillus species colonizing the honey bee gut. We serially passaged the four species through gnotobiotic bees and in liquid cultures in the presence of either pollen (bee diet) or simple sugars. Although the four species engaged in negative interactions, they were able to stably coexist, both in vivo and in vitro. However, coexistence was only possible in the presence of pollen, and not in simple sugars, independent of the environment. Using metatranscriptomics and metabolomics, we found that the four species utilize different pollen-derived carbohydrate substrates indicating resource partitioning as the basis of coexistence. Our results show that despite longstanding host association, gut bacterial interactions can be recapitulated in vitro providing insights about bacterial coexistence when combined with in vivo experiments.

Microbes colonize nearly every environment on Earth, from the ocean and soil to the inner and outer surfaces of animals, such as the gut or skin. They form communities that are usually made up of a diverse range of bacteria, often containing closely related species ­ a key factor for a successful community. But closely related bacteria can battle for the same resources, so it is unclear how they manage to live alongside each other without competing against one another. While diet is thought to play a key role in enabling closely related bacterial species to co-exist in the gut of an animal, experimental evidence is lacking, due to the difficulty in replicating these systems in the laboratory. One strategy for investigating microbial communities is using honeybees. A major dietary source for honeybees is pollen, which can also be applied in the laboratory to grow diverse types of bacteria found in the honeybee gut. In addition, scientists can generate bees that lack microbial communities in the gut, allowing them to add specific types of bacteria to study their impact. Brochet et al. used this approach with Western honeybees to assess whether diet enables closely related bacteria to live alongside one another in the gut. First, they colonized bees that lacked gut microbes with four closely related bacteria of the genus Lactobacillus, alone or together, and fed the bees either sugar water or sugar water and pollen. After five days, the gut bacteria were analysed. This revealed that bees fed on sugar water only had one dominant Lactobacillus species present in their gut, while bees fed with additional pollen harboured all four Lactobacillus species. Further analysis of these four bacterial species revealed that each of them activates distinct genes when grown on pollen, allowing the different species to consume specific nutrients from broken down pollen. These findings show that closely related bacteria can coexist in the gut by sharing the different nutrients provided in the diet of the host. Consequently, differences in dietary intake in honeybees and other animals may affect the diversity of gut bacteria, and potentially the health of an animal.