Organ Culture of Salivary Glands of Male Anastrepha suspensa (Diptera:Tephritidae)

Abstract

Salivary glands from male Caribbean fruit flies, Anastrepha suspensa, were isolated individually by dissection and cultured in Schneider's medium containing 10% Fetal Bovine Serum and antibiotics. A total of 103 salivary glands were successfully cultured. Two different procedures were used to evaluate viability. A trypan blue dye test revealed that no more than 10% of the cultured cells were non-viable, since they acquired blue coloration. In the second set of experiments, acid phosphatase activity of the cultured glands was measured colorimetrically from absorbance at 415 nm of liberated p-nitrophenol. This method indicated that the cultured glands stayed metabolically active, with enzyme activity equal to that of freshly excised glands on the fifth, eighth, and fifteenth days of the experiment. Scanning electron microscopy (SEM) showed no visual differences in tissue organization and size of individual cells when freshly excised salivary glands and cultured glands were compared. SEM showed that the cells of cultured glands were neither swollen nor shrunken, and close-up views showed no evidence of cell deterioration or lysing.