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Environmental sex-determination (ESD) is the phenomenon by which environmental factors regulate sex-determination, typically occurring during a critical period of early development. Southern flounder (Paralichthys lethostigma) exhibit temperature-dependent sex-determination that appears to be restricted to the presumed XX female genotype with the extremes of temperature, both high and low, skewing sex ratios toward males. In order to evaluate other environmental factors that may influence sex-determination, we investigated the influence of background color and cortisol on sex-determination in southern flounder. Experiments involving three sets of tanks, each painted a different color, were conducted at different temperatures using southern flounder of mixed XX-XY genotype. The studies involved rearing juvenile southern flounder in either black, gray, or blue tanks and sex-determination was assessed by gonadal histology. In both studies, blue tanks showed significant male-biased sex ratios (95 and 75% male) compared with black and gray tanks. The stress corticosteroid cortisol may mediate sex-determining processes associated with environmental variables. Cortisol from the whole body was measured throughout the second experiment and fishes in blue tanks had higher levels of cortisol during the period of sex-determination. These data suggest that background color can be a cue for ESD, with blue acting as a stressor during the period of sex-determination, and ultimately producing male-skewed populations. In a separate study using XX populations of southern flounder, cortisol was applied at 0, 100, or 300 mg/kg of gelatin-coated feed. Fish were fed intermittently prior to, and just through, the period of sex-determination. Levels of gonadal P450 aromatase (cyp19a1) and forkhead transcription factor L2 (FoxL2) messenger RNA (mRNA) were measured by qRT-PCR as markers for differentiation into females. Müllerian-inhibiting substance mRNA was used as a marker of males' gonadal development. Control fish showed female-biased sex ratios approaching 100%, whereas treatment with 100 mg/kg cortisol produced 28.57% females and treatment with 300 mg/kg cortisol produced only 13.33% females. These results suggest that cortisol is a critical mediator of sex-determination in southern flounder by promoting masculinization. This linkage between the endocrine stress axis and conserved sex-determination pathways may provide a mechanism for adaptive modification of sex ratio in a spatially and temporally variable environment.

An 18-week study was conducted in 12, 0.1ha ponds to evaluate the impacts of cyclic feeding regimes on hybrid striped bass (HSB) food¢sh production and pond water quality. Approximately 840 HSB [mean weight (std.); 91.08 g (8.18)] were stocked into each pond (8400 ¢sh ha À 1 ; 3360 ¢sh acre À 1 ) and fed according to one of three feeding regimes. The three feeding regimes included a control (fed twice daily to apparent satiation), and cycles of 3 weeks feed deprivation followed by 3 or 6 weeks of feeding to apparent satiation (3/3 and 3/6 respectively). Compensatory growth (CG) was observed in both cyclic feeding treatments; however, the response was insuf-¢cient for the ¢sh to completely regain lost weight. Final mean weight of control ¢sh (477.9 g) exceeded (Po0.05) that of ¢sh receiving the two cyclic treatments: 3/6 (404.7 g) and 3/3 (353.8 g). Speci¢c growth rate (SGR) of ¢sh in the 3/3 treatment increased during all three refeeding periods, and was signi¢cantly greater than controls during weeks 9^12 and weeks 15^18, which represent the refeeding phase of the second and third feeding cycles. Spe-ci¢c growth rate for ¢sh in the 3/6 treatment was signi¢cantly higher than controls only during the ¢rst 3 weeks of the ¢rst feeding cycle. Hepatosomatic index and condition factor were highly responsive measures that closely followed the metabolic state of ¢sh on the feeding cycle. Of the water quality variables measured, total phosphorus was 32% lower in ponds receiving cyclic feeding versus control ponds. Soluble reactive phosphorus was 41% and 24% lower in ponds o¡ered the 3/3 and 3/6 cyclic feeding treatments, respectively, although, signi¢cant di¡erences (Po0.10) were only observed between control and 3/3 treatment ponds. Overall, CG was observed in HSB food¢sh grown in ponds, although 3 weeks of feed deprivation was excessive and did not allow for complete growth compensation. Weight loss during feed deprivation was in£uenced by pond water temperatures. Early season feed deprivation did not cause as much weight loss as during the second cycle later in the season. Further studies on shorter deprivation periods applied during moderate to low water temperatures are needed to identify feeding regimes that minimize weight loss and result in a complete CG response.

Four separate studies were done on Southern flounder Paralichthys lethostigma larvae during first feeding and metamorphosis to determine the effects of stocking density, salinity, and light intensity on growth and survival. One study used stocking densities of 10, 20, 40, and 80 fish/L during first feeding; the second study compared the growth and survival of larvae stocked at 20 and 33 ppt; and a third experiment evaluated stocking densities of 1/L and 3/L under two different light intensities (1,600 lux vs 340 lux) during metamorphosis. The fourth experiment tested the effects of different salinities (0, 10, 20 and 30 ppt) on larval growth and survival during metamorphosis. Growth and survival (overall 6.9%) were not significantly different (P > 0.05) for stocking rates up to 80/L. Larvae placed into 20 ppt salinity had survival through first feeding similar to that of larvae raised at 33 ppt. During metamorphosis, light intensity had no effect (P > 0.05) on growth or survival, but fish stocked at 3/L had significantly lower (P < 0.05) survival than fish at 1/L. Complete mortality of larvae occurred at 0 ppt. Growth and survival past metamorphosis were not significantly different (P > 0.05) at 10, 20 and 30 ppt, but unmetamorphosed fish did not survive to day 60 at 10 ppt. Based on these results, practical larviculture of Southern flounder may require a two‐step process with high stocking rates (80 fish/L) through first feeding and lower densities (1/L) through metamorphosis. Fingerling production in fertilized nursery ponds might he possible at salinity as low as 20 ppt.

Southern flounder ( Paralichthys lethostigma ) exhibit environmental sex determination (ESD), where environmental factors can influence phenotypic sex during early juvenile development but only in the presumed XX female genotype. Warm and cold temperatures masculinize fish with mid-range conditions producing at most 50% females. Due to sexually dimorphic growth, southern flounder fisheries are dependent upon larger females. Wild populations could be at risk of masculinization from ESD due to globally increasing water temperatures. We evaluated the effects of habitat and temperature on wild populations of juvenile southern flounder in North Carolina, USA. While northern habitats averaged temperatures near 23 °C and produced the greatest proportion of females, more southerly habitats exhibited warmer temperatures (>27 °C) and consistently produced male-biased sex ratios (up to 94% male). Rearing flounder in the laboratory under temperature regimes mimicking those of natural habitats recapitulated sex ratio differences observed across the wild populations, providing strong evidence that temperature is a key factor influencing sex ratios in nursery habitats. These studies provide evidence of habitat conditions interacting with ESD to affect a key demographic parameter in an economically important fishery. The temperature ranges that yield male-biased sex ratios are within the scope of predicted increases in ocean temperature under climate change.

Compensatory growth (CG) or “catch‐up growth” is a period of super‐accelerated growth following a period of suboptimal conditions (i.e., lack of prey availability or overwintering). Little is known about the CG response in pond‐raised fish and whether hybrid striped bass (HSB), Morone chrysops × Morone saxatilis, might exhibit the rapid growth states or improvements in other production characteristics that may accompany the response. To evaluate the potential for CG in HSB culture, a 16‐wk growth trial in twelve 0.1‐ha earthen ponds was conducted. Approximately 2850 fish (mean weight ± SD = 3.2 g ± 1.1) were stocked into ponds and subjected to one of four cyclic feeding regimens. Treatment regimens included a control (0 wk, fed twice daily to apparent satiation) and cycles of 1, 2, or 4 wk of feed deprivation, followed by 1, 2, or 4 wk of feeding to apparent satiation. Fish in the 4‐wk feeding regimen were offered feed twice during the feed‐deprivation period (once every other week). Growth, specific growth rate (SGR), hepatosomatic index (HSI), intraperitoneal fat ratio, and condition factor (CF) were measured every other week, while overall growth, feed efficiency (FE) (FE = [weight gain/feed fed] × 100), and survival were calculated at the trial termination. The effect of these feeding regimens on water quality was examined by monitoring pH, turbidity, total ammonia nitrogen, nitrite–nitrogen, nitrate–nitrogen, soluble reactive phosphorus, and chlorophyll a weekly; total nitrogen and phosphorous biweekly; and dissolved oxygen and temperature twice daily. Cyclic feeding elicited CG; fish subjected to the 2‐wk regimen had a significantly higher SGR than 0‐wk controls during all but the final refeeding period. FE was higher for all fish in the cyclic regimens, although only FE for fish in the 2‐wk regimen was statistically greater (40%) than the controls. HSI was the most responsive measure and significantly decreased in the 2‐ and 4‐wk treatments during feed‐deprivation period and overcompensated during the refeeding period. CF also varied with feeding cycle and proved to be an effective nonlethal measure of predicting a CG response. No statistical differences in water quality parameters were observed. These data suggest that CG can be effectively induced in pond‐raised HSB and that the increase in FE warrants further research for practical application. Future pond studies with fingerling HSB fish should be conducted with emphasis on feed‐deprivation periods of 2 wk and refeeding periods of at least twice that of the feed‐deprivation period.

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