Abstract
Aspergillus fungi are ubiquitous inhabitants of colonies of the western honey bee (Apis mellifera), where they interact with bees in associations ranging from parasitism to possible mutualism. Aspergillus Flavi fungi are frequently found in bee bread (pollen processed for longterm storage) and are thought to contribute to food preparation, processing, preservation, and digestion. Conditions in the hive are challenging for fungi due, in part, to xeric and acidic properties of bee bread and the omnipresence of propolis, an antimicrobial product manufactured by bees from plant resins. We used quantitative and qualitative assays to determine whether A. flavus isolated from bee bread demonstrates tolerance for hive environmental conditions in terms of temperature, pH, osmotic pressure, and propolis exposure. Comparisons made use of three strains of A. flavus: a fungal biocontrol product not known from beehives (AF36), a strain isolated from bee bread (AFBB) in hives from central Illinois, and a pathogenic strain from a honey bee colony displaying symptoms of stonebrood (AFPA). Strain AFBB displayed higher tolerance of acidic conditions, low matric potential (simulating xeric substrate), and propolis exposure than did other strains. A genomic comparison between this new strain and the reference NRRL-3357 showed that AFBB, like AF36, might be blocked from carrying out aflatoxin biosynthesis. Sequence comparisons also revealed several missense variants in genes that encode proteins regulating osmotolerance and osmotic pressure in Aspergillus spp., including SakA, SskB, GfdA, and TcsB/Sln1. Collectively, results of our laboratory assays and genetic analyses are consistent with the suggestion that the strain isolated from bee bread is adapted to the bee bread environment and may have persisted due to a coevolutionary relationship between Aspergillus and A. mellifera. This finding bolsters recent concerns about the effects of fungicide use near bee colonies and broadens the ecological importance of highly adaptable fungal strains.