Abstract

One of the best indicators of colony health for the European honey bee (Apis mellifera) is its performance in the production of honey. Recent research into the microbial communities naturally populating the bee gut raise the question as to whether there is a correlation between microbial community structure and colony productivity. In this work, we used 16S rRNA amplicon sequencing to explore the microbial composition associated with forager bees from honey bee colonies producing large amounts of surplus honey (productive) and compared them to colonies producing less (unproductive). As supported by previous work, the honey bee microbiome was found to be dominated by three major phyla: the Proteobacteria, Bacilli and Actinobacteria, within which we found a total of 23 different bacterial genera, including known “core” honey bee microbiome members. Using discriminant function analysis and correlation-based network analysis, we identified highly abundant members (such as Frischella and Gilliamella) as important in shaping the bacterial community; libraries from colonies with high quantities of these Orbaceae members were also likely to contain fewer Bifidobacteria and Lactobacillus species (such as Firm-4). However, co-culture assays, using isolates from these major clades, were unable to confirm any antagonistic interaction between Gilliamella and honey bee gut bacteria. Our results suggest that honey bee colony productivity is associated with increased bacterial diversity, although this mechanism behind this correlation has yet to be determined. Our results also suggest researchers should not base inferences of bacterial interactions solely on correlations found using sequencing. Instead, we suggest that depth of sequencing and library size can dramatically influence statistically significant results from sequence analysis of amplicons and should be cautiously interpreted.

Highlights

  • As with other livestock producers, there is interest in the beekeeping industry as to whether it would be of benefit to feed probiotic bacteria to honey bee colonies

  • We show that Gilliamella strains in culture do not competitively inhibit the growth of honey bee specific bacteria, their dominance in the dataset correlated with a decrease in diversity and a greater ability to detect rare community members

  • Evolutionary analyses were conducted in MEGA6 using the Maximum Likelihood method based on the General Time Reversible Model with a gamma distribution, invariable sites, and 100 bootstrap replicates (Tamura et al, 2013). Foraging bees from both productive and unproductive colonies host many bacterial genera Illumina sequencing produced a large number of reads from the sampled honey bee colonies (3,252,621 raw reads and 2,903,512 post filtering), allowing a deep sampling of the honey bee microbial community (Fig. 1)

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Summary

Introduction

As with other livestock producers, there is interest in the beekeeping industry as to whether it would be of benefit to feed probiotic bacteria to honey bee colonies. We tested the hypothesis that the difference in productivity of colonies in the field was associated with a difference in the community structures of their respective gut microbiota These bacterial communities have been best characterized in worker bees, are consistently found across geography and season (Martinson et al, 2011), and are thought to consist of a core group of bacterial clades some of which have genus and species designations (Kwong & Moran, 2012; Engel, Kwong & Moran, 2013). This microbial community is thought to be socially transmitted within the hive via interaction with hive components and fecal material of congeners (Martinson, Moy & Moran, 2012; Powell et al, 2014) These communities are surprisingly consistent between workers and their increased prevalence during honey bee development would seem to suggest that these bacteria may contribute to the health of the bee. It is possible that the composition of the bacterial community in the honey bee may influence health and productivity

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