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The reproductive cycle of the rock oyster Striostrea prismatica was determined at two fishing areas, General Villamil and Ayangue (located on the southern coast of Ecuador), between May 2012 and April 2013. Monthly sampling campaigns were performed at both locations. The tissues were histologically examined to determine gonadal index (GI), oocyte development, follicular area coverage and sex ratio. Surface seawater temperature, salinity and chlorophyll a concentration were measured during samplings. Our results show a similar annual reproductive pattern at both locations. The GI reached maximum values during the summer. Oysters reached highest ripeness in January and February, while spawning occurred in February-March. Gametogenesis was linked to a consistently increasing follicular area (from 1.3 AE 0.7% to 85.8 AE 7.8%) and associated to surface seawater temperature. Spawning coincided with warm water temperature fluctuations and a seawater salinity decrease. No correlation was found with Chlorophyll a concentration. The sex ratio of sampled populations was 1:1, suggesting that oysters sizing more than 10 cm in shell length present a stable sex proportion in the population. The diameter of mature oocytes was significantly reduced (32.7%) during histologically preparations in comparison to fresh oocytes. Our study provides useful information of environmental factors that may control the observed gametogenesis and spawning activity of S. prismatica.