Stabilization of mineralized and sclerotized puparial cuticle of muscid flies

Calcium, magnesium and phosphorus are the major mineral elements in puparial exuviae of the face fly, Musca autumnalis, house fly, M. domestica and stable fly, Stomoxys calcitrans, but they are 20-50 times more prevalent in face fly than in the other two species that sclerotize the puparium. Carbon and nitrogen are approx. 5 times more abundant in house fly puparia than in face fly puparia. Face fly puparia contain two and three-fold less total amino acids than the house fly and stable fly, respectively. β-Alanine is a major amino acid in puparial cuticle of the house fly and stable fly, but it is absent in the face fly. There is no significant difference in glucosamine (chitin) content between the three species. Dopamine is the major catechol detected in face fly puparial cuticle while N-β-alanyldopamine (NBAD) is 10 to 15 times more prevalent than other catechols such as dopamine, N-acetyldopamine (NADA), 3,4-dihydroxyphenylalanine (DOPA) and 3,4-dihydroxyphenylacetic acid (DOPAC) in house fly and stable fly puparial cuticles. The latter two species have 75 to nearly 200 times higher levels of extractable catechols than the face fly. At the onset of pupariation, dopamine and NBAD attain nearly equivalent titres in puparial cuticles of face fly and house fly, respectively. Dopamine subsequently decreases more than 40-fold in the face fly as the cuticle becomes stabilized, while NBAD continues to accumulate in the house fly. The house fly covalently incorporates about 150 times more catechols in the puparium than does the face fly. The force required to fracture house fly and stable fly puparia is about three-fold greater than that required to fracture face fly puparia of comparable thickness. However, the face fly puparium attains a strength comparable to those of house fly and stable fly puparia by significantly increasing its thickness. These results demonstrate that dipterans use both catecholamines and minerals for stabilization of puparial cuticle with the house fly and stable fly relying primarily on sclerotization and the face fly on mineralization.