Increased understanding of the dry season survival mechanisms of Anopheles gambiae (An. gambiae) in semi-arid regions could benefit vector control efforts by identifying weak links in the transmission cycle of malaria. In this study we examine effect of seasonal indicators on morphologic and chemical characteristics known to contribute to suppression of water loss in mosquitoes. An. gambiae body size (indexed by wing length), mesothoracic spiracular index ((spiracle-length/wing-length)*100), and cuticular hydrocarbon (CHC) profiles were examined for their ability to differentiate mosquitoes exposed to aestivating and non-aestivating conditions in a laboratory setting. Mosquitoes exposed to aestivating conditions exhibited larger wing lengths, larger mesothoracic spiracular indices, and greater total CHC amount (standardized) than mosquitoes exposed to non-aestivating conditions. Total CHC amount increased with both mating and age. Mean n-alkane retention time (a measure of mean n-alkane chain-length) was lower in mosquitoes exposed to aestivating conditions, and increased with age. Individual CHC peaks were examined, and several CHCs were identified as potential biomarkers of aestivation, age, and insemination status. This study indicates that aestivation status of nulliparous female An. gambiae can be determined using both morphologic and chemical biomarkers.