A series of laboratory bioassays were conducted for the evaluation of the insecticidal efficacy of an isolate of Metarhizium anisopliae (Metschnikoff) Sorokin (Ascomycota: Hypocreales) against larvae of the Mediterranean flour moth, Ephestia kuehniella Zeller (Lepidoptera: Pyralidae), under various temperature–relative humidity (r.h.) conditions. The fungus was applied at four doses (0, 8 × 106, 8 × 108, and 8 × 1010 conidia ml−1) on wheat and insect mortality was assessed after exposure of 1, 2, 7, and 14 d. Bioassays were conducted at three temperatures (20, 25, and 30 °C) and two r.h. levels (55 and 75%). Although complete control was not achieved in any case, the fungus provided a considerable level of insect control. Mortality of E. kuehniella larvae on wheat treated with M. anisopliae ranged between 41.1 and 93.3% after 14 d of exposure, whereas the respective mortality levels in control dishes never exceeded 28.3%. The increase of temperature resulted in most cases to higher efficacy, indicating that temperature is an important factor for the performance of the fungus. In contrast, in most cases r.h. did not significantly affect the efficacy of the fungus, at least for the humidity levels tested.