Background The high invasiveness of phytophagous insects is related to their adaptability to various environments, that can be influenced by their associated microbial community. Microbial symbionts are known to play a key role in the biology, ecology, and evolution of phytophagous insects, but their abundance and diversity are suggested to be influenced by environmental stressors. In this work, using 16 S rRNA metabarcoding we aim to verify (1) if laboratory rearing affects microbial symbiont communities of Zeugodacus cucurbitae females, a cosmopolitan pest of cucurbitaceous crops (2) if temperature, diet quality, and antibiotic treatments affect microbial symbiont communities of both laboratory and wild populations, and (3) if changes in microbial symbiont communities due to temperature, diet and antibiotic affect longevity and fecundity of Z. cucurbitae. Results The results showed that microbial diversity, particularly the β-diversity was significantly affected by insect origin, temperature, diet quality, and antibiotic treatment. The alteration of gut microbial symbionts, specifically Enterobacteriaceae, was associated with low fecundity and longevity of Z. cucurbitae females feeding on optimal diet only. Fecundity reduction in antibiotic treated females was more pronounced when flies were fed on a poor diet without protein. Conclusions Our study proves the relationship between gut microbiome and host fitness under thermal and diet fluctuation highlighting the importance of microbial community in the adaptation of Z. cucurbitae to environmental stress.