Macro and microscopic maturation stage key of green crab (Carcinus maenas, Linnaeus 1758): Reproductive cycle and differences among estuarine systems

Gonadal histology is a helpful tool to validate species' macroscopic maturity stages in fisheries biology. Regardless of the many studies on Carcinus maenas, there are still few concerning gonads tissue histology and description of maturity stages. In Portugal, information regarding this species' biology to help support the regulation of crab fisheries, including the minimum landing size, has not yet been defined. In this work, the macro and microscopic characteristics of the reproductive cycle of C. maenas, oogenesis, and spermatogenesis development stages along the Portuguese coast are described, and a new macroscopic scale based on the histological analyses is suggested. During 2019 and 2020, adult C. maenas (both males and females) were collected from the Ria de Aveiro estuary, Rio Sado estuary, and Ria Formosa lagoon, respectively, North, Center, and South Portugal. No-significant differences (P > 0.05) were observed in carapace width and individual weight between all systems. Significant differences (P < 0.05) were observed in gonad weight, gonadosomatic index, and Fulton's condition index between the Rio Sado estuary and Ria Formosa lagoon. Furthermore, significant differences (P < 0.05) in the oocyte diameter between the northern and southernmost locations suggest a geographic variation related to the local environmental conditions of each system. Principal component analysis (PCA) revealed maturity stage and carapace width an association, and a similar morphometry between Ria de Aveiro and Ria Formosa. Temperature was correlated to both maturity stage and carapace width in Ria de Aveiro and Ria Formosa. From the histological analysis and based on microscopic criteria, it was observed that females previously classified within the late macroscopic development stage 2 should be considered mature, so a classification change in the current ovary's developmental stage is proposed. These findings can allow fisheries researchers to reclassify the estimations of maturity ogives and help support the regulation of this species' fishery. The proposed macroscopic scale was validated by histological analyses and can be used elsewhere.