AbstractInsect decline is a major threat to ecosystems around the world as they provide many important functions, such as pollination or pest control. Pollution is one of the main reasons for the decline, alongside changes in land use, global warming, and invasive species. While negative impacts of pesticides are well‐studied, there is still a lack of knowledge about the effects of other anthropogenic pollutants, such as airborne particulate matter, on insects. To address this, we exposed workers of the bumblebeeBombus terrestristo sublethal doses of diesel exhaust particles (DEPs) and brake dust, orally or via air. After 7 days, we looked at the composition of the gut microbiome and tracked changes in gene expression. While there were no changes in the other treatments, oral DEP exposure significantly altered the structure of the gut microbiome. In particular, the core bacteriumSnodgrassellahad a decreased abundance in the DEP treatment. Similarly, transcriptome analysis revealed changes in gene expression after oral DEP exposure, but not in the other treatments. The changes are related to metabolism and signal transduction, which indicates a general stress response. Taken together, our results suggest potential health effects of DEP exposure on insects, here shown in bumblebees, as gut dysbiosis may increase the susceptibility of bumblebees to pathogens, while a general stress response may lower available energy resources. Those effects may exacerbate under natural conditions where insects face a multiple‐stressor environment.