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We investigated the effects of fadrozol, an aromatase inhibitor (AI), and 17α-methyltestosterone (MT) on the induction of sex change in juvenile longtooth grouper Epinephelus bruneus, via histological observation of gonads. Changes in the mRNA expression of GtH subunits (FSH-β and LH-β) in the pituitary, and estradiol-17β (E2) and 11-ketotestosterone (11-KT) levels in the blood were also surveyed after AI and MT treatment. Juvenile longtooth groupers (113±17 g body weight; 16.2±1.2 cm body length) received intramuscular injections of AI at 3 (3-AI) and 5 (5-AI) mg/kg BW doses and MT at a 5 mg/kg BW (5-MT) dose. At week 7 post-injection, 3-AI and 5-MT oocytes were degenerated, and gonads of the 5-AI group initiated spermatogenesis. At week 21 post-injection, 3-AI- and 5-MT-treated gonads contained spermatogonia and spermatocytes, while 5-AI treatment induced advanced stages of spermatogenesis. The serum E2 level showed no significant differences throughout the experimental period, whereas that of 11-KT was significantly elevated in the 5-AI group at weeks 7 and 21 post-injection. A significant increase in the expression of FSH-β mRNA was evident in the 5-AI group at week 21 post-injection. In contrast, LH-β mRNA expression did not significantly differ among groups during the experimental period. These results imply that sex change has two stages in the longtooth grouper. In the first stage, oocytes are degenerated by the stimulation by 11-KT, and in the second stage spermatogenesis occurs, owing to the co-effects of 11-KT and FSH-β. [ABSTRACT FROM AUTHOR]