Abstract Background Game theory predicts that as the operational sex ratio (OSR; the ratio of sexually active males to fertilizable females) increases, reproductive males may adopt alternative reproductive behaviors. Empirical studies indicated that alternative behaviors may differ for different species. In the present study, we investigated the influence of different male-biased OSRs on male reproductive behaviors shown by a freshwater fish, the rose bitterling Rhodeus ocellatus. Results Six levels of the OSR (1:1, 2:1, 3:1, 5:1, 7:1, and 9:1) were established, and male leading, aggression, and preoviposition ejaculation behaviors were recorded. Our experiments showed that at low OSRs, the leading behavior of male rose bitterling increased with an increasing OSR, reaching a maximum at an OSR of 3:1, and then decreased as the OSR further increased. In contrast, aggression and preoviposition ejaculation behavior continually increased with an increasing OSR. Notably, males possibly lost their territorial position to other males and became sneakers when facing more rivals. Conclusions These observations suggest that at lower OSRs, male rose bitterling mainly rely on leading behavior, while at higher OSRs, they increasingly adopt aggression and preoviposition ejaculation. Therefore, for the rose bitterling, (1) decreased leading behavior and (2) increased aggression and preoviposition ejaculation are two alternative responses in the face of an increasing OSR, which is a more straightforward strategy for facing an increased risk of sperm competition than that adopted by its congener, the European bitterling.
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