Detrimental effects of thiamethoxam on the physiological status, gut microbiota, and gut metabolomics profile of Propsilocerus akamusi chironomid larvae (Diptera: Chironomidae)

Thiamethoxam, a widely applied neonicotinoid pesticide, poses a non-negligible risk to aquatic organisms and has garnered considerable attention. The biological impacts of thiamethoxam on chironomid larvae and protective strategies for tolerance remain to be investigated. In this study, we addressed the functional role of gut microbiota and determined the potential effects of thiamethoxam on physiological status, microbial commensals, and gut metabolome profile. A disturbed physiological status was induced by semi-lethal and sub-lethal thiamethoxam, with a higher concentration resulting in a more rapid and stronger response, as reflected by a conspicuous alteration of detoxifying and oxidative markers. Our results also demonstrated that an intact gut microflora was necessary for chironomid larvae to survive better under thiamethoxam-challenged condition. A low dosage of thiamethoxam could remarkably decrease the relative abundance of beneficial bacterial strains (e.g. Cetobacterium and Tyzzerella) while significantly increase the prevalence of opportunistic pathogens, including the genera Serratia, Shewanella, Aeromonas and Pseudomonas. Additionally, an evident variability of bacterial correlations was observed, and the thiamethoxam exposure impaired the genus-genus interaction and destabilized the whole community structure. The metabolome profile revealed that the toxic factor induced a significant downregulation of metabolites involved in glycolysis, amino acid metabolism and fatty acid metabolism pathways. Notably, the integration of metabolomics and gut microbiota data highlighted that representative substrates related to energy metabolism were negatively correlated with the elevated opportunities pathogens when chironomid larvae were challenged with thiamethoxam. These results suggested that a balanced microbial community was pivotal for maintaining energy expenditure and intake system, thus conferring benefits for chironomid larvae to defend against the invading thiamethoxam and preserve their physical well-being. This work provides theoretical guidance for the practical use of thiamethoxam in aquatic ecosystem and offers insights into the potential mechanisms utilized by chironomid larvae to detoxify pesticides.