Abstract Changes in water temperature and its seasonal timing influences the physiological processes of many aquatic ectotherms. Wastewater treatment plants (WWTP) contribute to warmer and more consistent water temperatures in streams draining the North American Great Plains, particularly during winter months. Reduced variation in seasonal water temperature may adversely affect fishes that rely on temperature fluctuation or specific overwintering temperatures for reproductive cues. We used johnny darter Etheostoma nigrum (Family Percidae), a thermally sensitive species native to the South Platte River Basin, as a focal species to assess the effects of winter water temperature regulations in Colorado. Our objective was to evaluate the effects of winter stream temperature and winter duration on reproductive success of johnny darter in the laboratory to inform development of water quality criteria for the South Platte River Basin. Winter duration (60, 90 or 120 days) and temperature (4 or 12°C) were crossed in a factorial design of six treatments that simulated warmed effluent‐impacted streams as well as streams with a more natural thermal regime. We defined reproductive success in johnny darter by examining four reproductive endpoints: (1) spawning timing, (2) egg and larvae production, (3) egg success, and (4) egg development. Across the four endpoints, seven main response variables were measured: spawning initiation date, eggs per gram of female per week, live larvae per gram of female per week, fertilisation success, hatching success, egg‐to‐larva survival and days to hatch. Winter temperature and duration influenced spawning timing and egg development. Earlier spawning initiation dates were observed in fish exposed to 12°C winters and was associated with a higher number of days to hatch. Eggs per gram of female per week, live larvae per gram of female per week, fertilisation success, hatching success, and egg‐to‐larva survival were similar among winter treatments. Although the State of Colorado chronic winter water temperature standard of 12°C appears to be adequate for egg and larvae production in johnny darter, our results showed that fish exposed to the standard began spawning significantly earlier than fish exposed to 4°C winters. Evaluating reproductive output in the context of seasonal timing is valid because spawning timing has the potential to affect overall production and egg development, and consequently recruitment and population sustainability. Therefore, spawning timing should be considered when making management decisions and revising water temperature standards. The warming of streams and rivers from WWTP effluent is a global issue, especially in areas that are undergoing rapid urbanisation. Fishes are particularly sensitive to water temperature and many fishes worldwide are likely to be experiencing changes in important physiological processes, affecting both reproduction and survival. Our study indicates that reproductive timing could be influenced by stream warming and stresses the need for further studies on the influence of anthropogenic effects on stream warming.
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