Bacterial secreted products selectively inhibit non-symbiotic fungi in bees

Abstract

Microbial interactions play an important role in shaping microbiome assembly, such as by limiting invasion by harmful organisms that can directly affect the host or disrupt microbiome-associated benefits. Such interactions have been observed across systems, including in the microbiomes of key pollinators such as stingless bees. In Scaptotrigona depilis, bacteria associated with the larval diet inhibit potentially pathogenic filamentous fungi while allowing beneficial yeast symbionts to persist. The mechanisms underlying these effects, however, remain unclear. Here, we combined conditioned media (cell-free supernatant) assays with genomic and metabolomic analyses to investigate whether bacterial secreted products mediate these effects in the bee microbiome. Our results show that bacterial secreted products, particularly from prevalent bacterial taxa such as Apilactobacillus kunkeei, strongly affect fungal growth. Filamentous fungi, including the pathogen Aspergillus, were consistently inhibited, partly through substrate acidification driven by organic acids, but also through additional acidity-independent factors. In contrast, yeast responses were more variable: a non-symbiotic Zygosaccharomyces was inhibited by bacterial metabolites under near-neutral pH, whereas the symbiotic Zygosaccharomyces required for larval development was maintained or promoted under acid-conditioned media. Genomic analyses revealed limited canonical antifungal biosynthetic clusters in the most prevalent bacteria in the larval diet, while metabolomics identified extracellular peptide-like compounds across strains, suggesting a role for non-canonical secreted products. Together, these results show that bacterial secreted products play a key role in selectively shaping fungal communities in the stingless bee larval diet, providing first hints on a mechanistic basis for how microbial interactions structure this ecosystem.