Bradysia odoriphaga and Bradysia difformis are devastating pests of vegetable, ornamental crops and edible mushrooms causing significant losses. Temperature may be an important factor restricting their population abundance in the summer. To determine the effects of short-term heat shock on adults, their survival, longevity and fecundity data were collected, and antioxidant responses and heat shock protein expression levels were examined. Our results indicated that the survival rates of Bradysia adults decreased rapidly after heat shock ≥36 °C, and the longevity and reproductive capacities were significantly inhibited, indicating that short-term heat shock had lethal and sub-lethal effects. Moreover, the lipid peroxidation levels of B. difformis and B. odoriphaga increased dramatically at 36 °C and 38 °C, respectively. Four antioxidant enzymes activities of B. odoriphaga were greater than those of B. difformis at 38 °C. Additionally, hsp70 and hsp90 expression levels significantly increased after heat stress, and higher expression levels of B. difformis and B. odoriphaga were discovered at 36 and 38 °C respectively, indicating their different heat tolerance levels. Overall, short-term heat shock (≥36 °C) caused significantly adverse effects on Bradysia adults, indicating that it could be applied in pest control, and antioxidant system and hsp genes played important roles in their heat tolerance levels.