Polo, R. O. | bioRxiv Science

bioRxiv2026

Honey bee (Apis mellifera) gut microbiome associations with viruses and pesticides across Canadian agroecosystems

The honey bee (Apis mellifera) gut microbiome plays a central role in host health, yet its variation across agricultural landscapes remains poorly resolved. This study investigates how major environmental stressors, particularly pesticide exposure and RNA virus loadings, shape the honey bee gut microbiome in a large-scale field study conducted across Canada, spanning diverse agroecosystems from British Columbia to Quebec. We identify consistent associations between specific bacterial taxa and major RNA viruses, including enrichment of Serratia marcescens with SBV and depletion of Bombella intestini with BQCV. Pesticide exposure is likewise linked to reproducible shifts in key microbial taxa. Together, these findings reveal that interacting stressors jointly shape the bee gut microbiome and enable prediction of microbiome responses in agroecosystems. HighlightsDistinct associations identified between gut bacteria and major bee RNA viruses (BQCV, SBV, LSV, IAPV) Pesticide exposure is linked to reproducible shifts in key microbial taxa Combined virus-pesticide effects form coordinated clusters that predict microbiome variation and specific bacterial responses Integrated modeling demonstrates that environmental stressors can jointly explain microbiome structure beyond crop effects Graphical abstractSchematic overview of potential links between pesticide exposure and RNA virus infection and their effects on the bee gut bacterial community. Solid arrows indicate associations supported by the present study, whereas dashed arrows indicate hypothesized or unresolved interactions. Associations between the presence of specific bee RNA viruses (left) or pesticide residues (right) and changes in the relative abundance of particular gut taxa (pink {uparrow}, increased; blue {downarrow}, decreased). The pesticide subtype is indicated by the icon in the cell (leaf - herbicide, hyphae - fungicide and insect - insecticide). Several bacterial taxa showed reproducible associations with specific viral or pesticide variables, including Bombella intestini, Serratia marcescens, Melissococcus plutonius, Paenibacillus alvei, Apibacter sp. wkB309, and Gilliamella sp. A7. Abbreviations: BQCV Black queen cell virus; LSV, Lake Sinai virus; SBV, Sacbrood virus; IAPV, Israeli acute paralysis virus. (p/n/b) indicate the sample matrix in which the pesticide was detected, namely pollen, nectar, and bee tissue, respectively. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/731697v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@cb92a4org.highwire.dtl.DTLVardef@1087045org.highwire.dtl.DTLVardef@102cabforg.highwire.dtl.DTLVardef@4ce2f1_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2026

Comparative Proteomics Across Tissues and Crop Agroecosystems Reveals Agricultural Stressor Responses in the Western Honey Bee

Maintaining honey bee health in crop production systems is increasingly difficult because worker bees encounter multiple chemical and biological pressures from pesticides and pathogens. How these field-realistic pressures affect molecular physiology across functionally distinct tissues remains poorly understood. Here, we tested whether tissue-resolved proteomics could separate stable tissue-specific patterns from crop-associated molecular changes. To do this, we profiled abdomen, gut, and head proteomes from honey bees collected across four Canadian crop ecosystems over two consecutive years, and integrated these data with pesticide-residue and pathogen-load measurements. Proteomic variation was structured by both tissue identity and crop environment. Tissue-specific proteomic profiles were characterized across samples, whereas crop-associated effects were detected in both years and were stronger in 2021, the second year of the study. Tissue-specific enrichment and network analyses linked the abdomen to lipid catabolism and ubiquitin-proteasome proteostasis, the gut to central carbon metabolism, membrane transport, vesicle trafficking, and cytoskeletal organization, and the head to neurosensory and mitochondrial functions, together with amino-sugar metabolism and vesicle-associated quality-control modules. Among the measured pesticide residues, boscalid was the most reproducible chemical correlate of proteomic variation, with the strongest signal in the gut. Cross-year validation associated boscalid exposure with reduced abundance of gut proteins involved in mitochondrial metabolism, protein quality control, vesicle trafficking, nutrient transport, and biosynthetic pathways. Additionally, integrated proteome-transcriptome-microbiome factor analysis further identified gut-centered components associated with measured stressor variables and linked protein-level variation to coordinated transcriptomic and microbial shifts. Independent-year validation showed that compact crop-associated protein signatures detected in 2020 were also present in 2021. Together, these results show that honey bee tissues maintain stable proteomic identities while showing tissue- and year-specific responses to pesticide and pathogen pressures encountered in crop ecosystems. The gut proteome may specifically provide a sensitive molecular indicator of pesticide-associated perturbation under field conditions.

bioinformatics↗