Diapausing pupae of the flesh fly Sarcophaga crassipalpis (Diptera: Sarcophagidae) are more resistant to inoculative freezing than non‐diapausing pupae

Summary The resistance of diapausing (overwintering) and non‐diapausing (summer) Sarcophaga crassipalpis (Diptera: Sarcophagidae) pupae to inoculative freezing was examined. Although both types of pupae resisted inoculative freezing after 24‐h submergence in water, diapausing pupae were overall significantly more resistant than non‐diapausing pupae. Exposing the thin pupal cuticle by removing the ends of the puparial case eliminated the capacity of both pupal types to resist inoculative freezing, indicating that resistance to inoculative freezing resides with the puparium. Pupae submerged in surfactant solution were significantly less resistant to inoculative freezing than those submerged in water. Furthermore, the puparial water content of pupae submerged in surfactant solution was significantly greater than that of puparia from pupae submerged in water. Surfactant may have promoted inoculative freezing by facilitating the spread of water over the surface of and into the puparium, thereby creating bridges between external ice and pupal body fluids. Extracting puparial surface lipids with chloroform/methanol (2 : 1, v:v) decreased the resistance of non‐diapausing pupae to inoculative freezing but did not significantly affect that of diapausing pupae. This finding indicates that the puparium of diapausing pupae contains protection against inoculative freezing separate from its surface lipids. This barrier may be important in protecting the freezing‐intolerant overwintering pupae against inoculative freezing within their soil hibernaculum.