Abstract  [Aim]  Gut microbes can directly and indirectly affect host health. Several studies have shown that gut microbes can improve egg production, but the mechanisms involved remains unclear. This study aims to determine how gut microbial metabolites influence oviposition in bumblebees. [Methods]  Non-target metabolomics analysis was used to compare the relationship between intestinal metabolites and oviposition in Bombus terrestris and B. lantschouensis queen bees. Six laying queens and six females that were non-ovipositing for 28 d were selected from each species and metabolomics analysis was performed on their intestinal contents. [Results]  Enrichment analysis indicated significant enrichment in the lipid metabolism pathways in queens from both the Bombus terrestris (DQA1) and B. lantschouensis (LQA1) ovipositing groups. The cysteine and methionine metabolic pathways were significantly enriched in the Bombus terrestris ovipositing (DQA1) and 28 d non-ovipositing (DQW) groups. Whereas, for B. lantschouensis, the sphingolipid metabolism pathway was significantly enriched in both the LQA1 and 28 d non-ovipositing (LQW) group. There were no significantly enriched metabolic pathways in the 28 d non-ovipositing B terrestris (DQW) or B. lantschouensis (LQW) groups. [Conclusion]  There was a significant difference in lipid metabolism pathways between B. terrestris and B. lantschouensis queen bees. Non-ovipositing B. terrestris lacked the L-Cysteine and L-Methionine metabolites in the Cysteine and methionine metabolism pathways. The O-Phosphoethanolamine metabolites in the Cysteine and methionine metabolism pathways were absent in non-ovipositing queen bumblebees.