Combined effects of photoperiod and salinity on growth,survival, and osmoregulatory ability of larval southern flounder Paralichthys lethostigma

The southern flounder, Paralichthys lethostigma, is an important commercial and recreational marine flatfish that inhabits estuaries and shelf waters in the south Atlantic, from North Carolina through the Gulf coasts, with the exception of south Florida. Because juvenile and adult fish are highly euryhaline, it is a prime candidate for aquaculture. Methods for captive spawning of southern flounder are well developed; however, information on optimal culture requirements of the early larval stages is required for reliable mass production of juveniles.To determine the optimal photoperiod and salinity conditions for culture from hatching to day 15 post-hatching (d15ph), embryos were stocked into black 15-l tanks (75 l⁻¹) under four photoperiods (24L:0D, 18L:6D, 12L:12D, and 6L:18D) and two salinities (25 and 34 ppt) in a 4×2 factorial design. Temperature was 18 °C, light intensity was 150 lx, and aeration was 50 ml min⁻¹. Significant (P<0.05) effects of photoperiod and salinity on growth (notochord length, wet and dry weights) were obtained. Growth increased with increasing photoperiod and salinity and was significantly greater at 24L and 18L than at 12L or 6L, and at 34 than at 25 ppt. On d11ph and d15ph, significant interactive effects between photoperiod and salinity on growth (wet and dry weights) were also evident. Growth of larvae reared at 25 ppt increased with increasing photoperiod to a maximum at 24L, while growth of larvae at 34 ppt reached a plateau at 18L. While there were no significant photoperiod effects on these parameters, larval survival, body water percentage, and larval osmolality on d15ph were significantly higher at 34 than at 25 ppt (41% vs. 16% survival; 322 vs. 288 mosM kg⁻¹; and 84% vs. 76% water, respectively), suggesting stress and nonadaptation to 25 ppt, a salinity more nearly isoosmotic than full-strength seawater. Since larvae from both salinity treatments were neutrally or positively buoyant at 34 ppt, but negatively buoyant at 25 ppt, larvae reared at 25 ppt probably allocated energy to maintain vertical positioning, compromising growth and survival.The results demonstrate that growth and survival of early-stage southern flounder larvae are maximized under long photoperiods of 18–24L and in full-strength seawater. Longer photoperiods probably extend the time larvae have for feeding, while full-strength seawater salinity optimizes buoyancy and vertical positioning, conserving energy. The results show that early larval stage southern flounder larvae are not entirely euryhaline, which involves not only the ability to osmoregulate, but to conserve energy under reduced buoyancy. This is consistent with suboptimal vs. maximal growth of larvae reared at 25 and 34 ppt, respectively, under 18L (i.e., photoperiod×salinity interaction). This is also consistent with other reports that tolerance to lower salinities in these euryhaline flatfish increases post-metamorphosis when transition from a pelagic to benthic existence alleviates the need to counteract reduced buoyancy.     

Effects of salinity, aeration and light intensity on oil globule absorption, feeding incidence, growth and survival of early-stage grouper Epinephelus coioides larvae

ABSTRACT: A series of experiments were conducted to examine the effects of salinity, aeration and light intensity on oil globule absorption, feeding incidence, and growth and survival of early-stage Epinephelus coioides larvae. Newly hatched larvae were transferred to 40-L aquaria at a density of 1500 individuals/aquarium. Larvae were exposed to different levels of aeration (0 mL/min per L, 0.62 mL/min per L, 1.25 mL/min per L, 2.50 mL/min per L, or 3.75 mL/min per L); salinity (8 ppt, 16 ppt, 24 ppt, 32 ppt, or 40 ppt); and light intensity (0 lx, 120 lx, 230 lx, 500 lx, or 700 lx) for 4–6 days. Twenty larvae were sampled daily at 11:00 hours to measure for total length (TL), oil globule volume, and feeding incidence. Survival rates were determined by counting the total number of larvae remaining in each aquarium at the end of the experiment. Significantly higher survival rates ( P   <  0.05) were observed at aeration levels of 0.62 mL/min per L and 1.25 mL/min per L, at salinity levels of 16 ppt and 24 ppt, and at light intensities of 500 lx and 700 lx. The influence of aeration level, salinity and light intensity on oil globule absorption, feeding incidence, and growth and survival of early-stage grouper larvae are discussed.     

Effects of Light Intensity and Salinity on Growth, Survival, and Whole-Body Osmolality of Larval Southern Flounder Paralichthys lethostigma

The southern flounder Paralichthys lethosligma is a high‐valued flatfish found in estuarine and shelf waters of the south Atlantic and Gulf coasts of the United States. Wide temperature and salinity tolerances exhibited by juveniles and adults make it a versatile new candidate for commercial culture, and studies are underway in the southeastern U.S. to develop hatchery methods for this species. The objectives of this study were to establish illumination and salinity conditions that optimize growth and survival of larval southern flounder reared through the yolk‐sac and first feeding stages to 15‐d post‐hatching (15 dph). Early embryos were stocked into black 15‐L tanks under light intensities of 5, 50, 100, and 1,000 Ix and at salinities of 24 and 34 ppt in a 4 ± 2 factorial design. Significant (P 0.05) effects of both light intensity and salinity on growth and survival were obtained, with no interaction between these effects. On 11 dph and 15 dph, growth was generally maximized at the intermediate light intensities (50 and 100 Ix) and minimized at the extremes (5 and 1,000 Ix). By 15 dph, growth was higher at 34 ppt than at 24 ppt. Survival to 15 dph showed trends similar to those of growth. Survival was higher at 100 Ix (avg. = 46%, range = 41–54%) than at 5 Ix (avg. = 11%, range = 6–17%) and higher at 34 ppt (avg. = 43%, range = 3145%) than at 24 ppt (avg. = 17%, range = 8–38%). Whole‐body osmolality (mOsmol/kg) was significantly lower in larvae reared at 24 ppt (avg. = 304, range = 285–325) through 11 dph than in larvae reared at 34 ppt (avg. = 343, range = 296–405). Larvae reared under the extreme light intensity treatments (5 and 1,000 Ix) at 34 ppt appeared to exhibit osmoregulatory stress, particularly on 11 dph, when a marked increase in whole‐body osmolality was observed. The mid‐intensity treatments (50 and 100 Ix) at 34 ppt optimized growth and survival of larval southern flounder in this study; and elicited the most stable osmotic response. These conditions appear to be consistent with those that southern flounder larvae encounter in nature during this early developmental period.     

Effects of Stocking Density, Salinity, and Light Intensity on Growth and Survival of Southern Flounder Paralichthys lethostigma Larvae

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.     

Evaluation of First-Feeding Regimens for Larval Nassau Grouper Epinephelus straitus and Preliminary, Pilot-Scale Culture through Metamorphosis

Two 10-day hatchery experiments were conducted to evaluate s-type (Hawaiian strain) and ss-type (Thailand strain) rotifers Brachionus plicatilis and cryogenically preserved oyster Crassostrea gigas trochophores as first feeds for larval Nassau grouper Epinephelus striatus. Newly hatched grouper larvae were reared at densities of 11.2–20.8/L in 500-L tanks at 36–38 ppt salinity, 25–26 C, and under a 11-h light: 13-h dark photoperiod. Beginning on day 2 posthatching (d2ph), prey were maintained at a density of 20 individuals/mL, while phytoplankton (Nanochloropsis oculata) was maintained at 500 × 10³ cells/mL. In experiment 1, survival and growth were higher (P < 0.05) for fish fed small s-type rotifers (mean lorica length = 117 μm; fish survival = 7.96%) selected by sieving than for fish fed non-selected rotifers (mean lorica length = 161 μm; fish survival = 2.13%). These results demonstrated the advantage of small prey size and suggested that super-small (ss-type) rotifer strains would be beneficial. In experiment 2, three feeding regimens were compared: 1) ss-type rotifers (mean lorica length = 147 μm); 2) oyster trochophores (mean diameter = 50 μm) gradually replaced by ss-type rotifers from d5ph; and 3) a mixed-prey teatment of 50% oyster trochophores and 50% ss-type rotifers. Survival was higher (P < 0.05) for larvae fed mixed prey (15.6%) than for those fed rotifers (9.73%) or trochophores and rotifers in sequence (2.55%), which also showed the slowest growth. Oyster trochophores, although inadequate when used exclusively, enhanced survival when used in combination with rotifers, possibly by improving size selectivity and dietary quality. In a pilot-scale trial, larvae were cultured through metamorphosis in two 33.8-m³ outdoor tanks. Fertilized eggs were stocked at a density of 10 eggs/L and larvae were fed ss-type rotifers from d2ph-d20ph, newly hatched Artemia from d15ph-d18ph, 1-d-old Artemia nauplii from d18ph-d62ph. Survival on d62ph was 1.17%, with a total of 5,651 post-metamorphic juveniles produced.     

Effect of Salinity on Larval Rearing of Pacu,Piaractus mesopotamicus,a Freshwater Species

The aim of this study was to evaluate the growth and survival of pacu, Piaractus mesopotamicus, larvae reared in different salinities and to determine the Artemia nauplii life span in freshwater and in saline water. First feeding 5‐d‐old pacu larvae were reared in freshwater or at 2, 4, 6, 8, 10, 12, and 14 ppt salinities. The larvae were reared in 1.5‐L aquaria at a density of 10 larvae/L with three replicates per treatment. After 10 d of rearing, significant differences (P < 0.05) were observed for growth and survival. Larval growth was higher at 2 and 4 ppt, and survival at 2 ppt was 100%. In freshwater and at 4, 6 and 8 ppt, the survival was 91.1, 93.3, 73.3, and 39.9%, respectively. At higher salinities, there was 100% mortality after 2 h (12 and 14 ppt) and 8 h (10 ppt) of exposure. The slightly saline water of at least 2 ppt increased the Artemia nauplii life span compared to the life span in freshwater. Later, in a second trial, 5‐d‐old pacu larvae were reared in freshwater and at 2 and 4 ppt salinities during the first 5 or 10 d of active feeding, and then the fish were transferred to freshwater. At the end of 15 d, larval growth was lower in freshwater (42 mg) than in treatments 2 and 4 ppt (59–63 mg). The abrupt transfer of fish from freshwater to slightly saline water and the return to freshwater did not affect the survival rates (89–97%). The larvae were able to adapt to these saline environments and handle abrupt changes in salt concentration. We concluded that salinity concentration of 2 ppt can be used for pacu larval rearing, allowing the Artemia nauplii lifetime to last longer and cause faster fish growth.     

漠斑牙鲆仔鱼、稚鱼和幼鱼对低盐度的耐受力和淡水驯化技术的研究

研究了漠斑牙鲆仔鱼、稚鱼和幼鱼对低盐度的耐受力和漠斑牙鲆的淡水驯化技术。初孵仔鱼在盐度为5的水中最多存活5d,5d以后全部死亡。30日龄稚鱼在4h10min淡水组全部死亡,对淡水的耐受力较低,而盐度为5～25的各组72h的存活率都在95.00%以上。90日龄幼鱼经缓慢淡水驯化9d后存活率可达98%以上,对漠斑牙鲆幼鱼进行淡水驯化的最佳年龄为90日龄。淡水驯化影响幼鱼初期的摄食,以后则逐渐趋于正常;急性淡水驯化对幼鱼的伤害比缓慢淡水驯化大;漠斑牙鲆对低盐的耐受力随生长和发育逐渐增加,年龄是影响幼鱼对低盐度耐受力的主要因素。     

Nitrite Toxicity and Methemoglobin Changes in Southern Flounder,Paralichthys lethostigma,in Brackish Water

This study was performed to estimate the nitrite toxicity to southern flounder, Paralichthys lethostigma, in brackish water (7.5 ppt of salinity). For a LC₅₀ test, 20 fingerlings (5.7 ± 0.4 cm) in each aquarium (15 L) were exposed to the concentrations of 0, 1, 5, 10, 15, 30, 60, 120, and 240 mg NO₂^?‐N/L in duplication for 10 d. Median lethal concentration at 96 h (96‐h LC₅₀) was calculated as 81.6 mg NO₂^?‐N/L. For a verification test, young flounder (164.2 ± 9.1 g) were exposed to a simulated culture condition in recirculating systems (1000 L). Sodium nitrite was not added to control system, whereas it was added to Treatment system 1 (TS 1) and Treatment system 2 (TS 2) to maintain nitrite concentrations of 20 and 30 mg NO₂^?‐N/L, respectively. The plasma nitrite concentrations of the young flounder in TS 1 and TS 2 were 4.5 and 6.6 mg NO₂^?‐N/L, respectively, after 2 wk. At this time, the methemoglobin percentages in TS 1 and TS 2 reached 85.8 and 89.7%, and survival rates were 37.5 and 25.0%, respectively. The results of these tests indicate that southern flounder do not concentrate nitrite in blood from the environment, but they seem to be more sensitive to nitrite compared with other species that do not concentrate nitrite.     

Food Conditions and Water Salinity Affect Survival and Growth of Golden Mandarin Fish,Siniperca sherzeri,Larvae through Transcriptional Regulation of Growth and Lipometabolic Genes

Failing to initiate first feeding during the transition from endogenous nutrition to exogenous feeding will lead to starvation of fish larvae. However, little is known about the mechanism of first feeding selection of fish. Golden mandarin fish larvae (3 d after hatch, 2.05 ± 0.03 mg) were fed with four different foods for 7 d, including the following: M – Megalobrama amblycephala (prey fish larvae as natural food); S – surimi of M. amblycephala; A – Artemia (zooplankton); and MA –mixed M. amblycephala with Artemia (mixed food). Larvae fed with the mixed food achieved an appropriate balance between high survival and good growth through elevating the expression of growth genes (GH, IGF‐I, and IGF‐II) and fatty acid synthesis genes (FAD and ELO). Growth performance of fish fed with MA reared at different salinities (0, 5, and 10 ppt) was examined. The salinity of 5 ppt produced the best growth performance of the three salinity levels tested. Fish larvae adapted to high‐ or low‐salinity environments through increasing the expression of lipolysis genes (HSL, LPL, and HL). Therefore, both food type and salinity affect the growth, survival, and lipometabolism of golden mandarin fish larvae during initial feeding stage, and mixed food and 5 ppt salinity improved its survival and growth.     

Salinity Effects on Early Life Stages of Southern Flounder Paralichthys lethostigma

Abstract.— In South Carolina, studies have been conducted to develop rearing techniques for southern flounder Paralichthys lethostigma a candidate for aquaculture development and stock enhancement programs. To help define environmental tolerances, a variety of salinity studies were conducted with the early life stages of this species. Eggs were buoyant at 32 ppt and sank at 29 ppt with salinities of 30–31 ppt providing varying levels of suspension in the water column. Eggs incubated at 0 and 5 ppt all died, whereas 82.5% hatched at 10 ppt but larvae died shortly thereafter. At 63 h post-fertilization, there were no differences in hatch level for eggs incubated at salinities of 15 to 35 ppt (mean hatch level 98.5%). In a 72-h study, fish 3 wk post-metamorphosis (13.7 mm TL, 50-d-old) were acclimated to seven salinities ranging from 0–30 ppt. Fish held at 0 ppt salinity exhibited a statistically (P < 0.05) lower survival (20.0%) than those exposed to 5–30 ppt salinity concentrations. No differences were detected in survival (mean 99.1%) among fish held in the higher salinities. A second study examined the tolerance of older juveniles to lower salinities. Juvenile flounder (95.2 mm TL, 220-d-old) were acclimated to 0, 1,5 and 10 ppt salinities and reared for 2 wk. Results showed that fish could tolerate salinities of 0–10 ppt (100% survival). These data indicate that salinity tolerance of southern flounder increases with age. In addition to the short duration studies, a replicated 11-mo duration tank grow-out study was conducted at mean salinity 5.4 ppt and mean temperahue 22.6 C with an all male population. Flounder grew from a mean length of 100 mm to 213 mm TL and weight from 8.9 to 104.3 g. Growth of the cultured fish approximated that observed among male flounders in the wild.     

Metamorphosis in summer flounder: manipulation of rearing salinity to synchronize settling behavior, growth and development

In the aquaculture of summer flounder (Paralichthys dentatus), the inherent variation in growth and settling behavior during metamorphosis may lead to cannibalism and necessitate increased labor due to size grading. Our goal was to use an environmental salinity change as a cue to synchronize settling behavior and produce a larger, more uniformly sized cohort of juvenile summer flounder. Early metamorphic flounder were exposed to either a 5-day fluctuating (30–20–30–20–30 ppt; “Flux”) or a single (30–20 ppt; “20 ppt”) drop in rearing salinity. A control (continuous 30 ppt) was used for comparison. Average values for a peak settlement interval (PSI; defined as the interval beginning on the day by which the first 20% had settled until the day 80% had settled) were not affected by salinity manipulation, though the 20-ppt treatment did significantly increase percent settled per day by 54 dah. Average fish size was increased by the 20-ppt treatment (19.3±0.5 mm), but not the flux treatment (17.2±0.4 mm), compared to the control (17.6±0.5 mm). Developmental stage was significantly increased in the 20-ppt treatment (3.2±0.1) in comparison to the Flux (2.9±0.1), but not the control (3.1±0.1). However, the 20-ppt treatment reduced variance in development. To confirm the positive effects of the 20-ppt treatment, a second experiment was performed. A single salinity drop (“20 ppt”), a previously successful ([Gavlik, S., Albino, M., Specker, J.L., 2002. Metamorphosis in summer flounder: Manipulation of thyoid status to synchronize settling behavior, growth, and development. Aquaculture 203 (3-4), 359-373]thyroid hormone manipulation treatment (“TH”) and a combination of the two (“TH+20 ppt”) were compared to a control (continuous 30 ppt, no thyroid manipulation). The mean PSI was significantly reduced by both TH (7±1 days) and TH+20 ppt (8±0 days) treatments, compared to the control (11±1 days). The PSI for the 20-ppt treatment (9±0 days) was not significantly different than the control. The percent flounder settled per day was significantly increased by 20 ppt salinity and significantly modified (decreased, then increased) by TH manipulation. Flounder exposed to 20-ppt salinity were both larger (20 ppt: 18.6±0.3 mm; TH+20 ppt: 18.3±0.3 mm) and more developmentally advanced (20 ppt: 3.1±0.04; TH+20 ppt: 3.2±0.03) than flounder in 30 ppt (TH: 17.8±0.3 mm/3.1±0.3; Control: 17.9±0.3 mm/3.0±0.05). Finally, 20-ppt treatment reduced variance in development, while TH treatment reduced variance in both growth and development. Percent survival was unaffected by treatment in both experiments. In summary, a decrease in rearing salinity, from 30 to 20 ppt, increased growth, settling behavior and development of metamorphosing summer flounder. A decrease in rearing salinity, in combination with a TH manipulation, should result in larger, more uniformly sized flounder cohorts. We expect this synchronization will reduce the cannibalism and labor costs associated with size grading of cultured, metamorphosing summer flounder.     

Changes in Plasma Cortisol, Glucose, and Selected Blood Properties in the Summer Flounder Paralichthys dentatus Associated with Sequential Movement to Three Experimental Conditions

To determine the changes in blood chemistry associated with sequential transfer of summer flounder Paralichthys dentatus (320–480 g), 300 hatchery-reared fish were moved to three different environmental conditions during a 20-d period. Fish were transferred in progression from a recirculating seawater system (22 ppt, 22.5 C) to a flow-thru seawater system (31 ppt, 20.0 C), to three small coastal net pens (33 ppt, 15.5 C), and finally to a large open ocean net pen (33 ppt, 16.0 C). For this study, eight random fish were captured at each progressive step (environmental condition), anesthetized (MS222), and bled from the caudal vein (2 mL). Transferred flounder were bled every 12 h for 48 h to collect plasma cortisol and glucose samples. Fish were bled 24 h after transport and every 3 d thereafter for osmolarity, hematocrit, hemoglobin concentration, mean corpuscular hemoglobin content, glucose, cortisol, and the electrolytes Cl^- Na⁺, K⁺ and Ca⁺. The most significant perturbations to blood chemistry (P < 0.05) occurred within 24 h of initial transfer from the recirculating to flow-thru seawater systems, suggesting an osmoregulatory rather than handling or transfer related stress. Osmolarity, electrolyte, and hematological parameters fluctuated and then recovered to stable levels by day 8 in the flow-thru seawater system. However, unlike the initial transfer, successive movement to the coastal and then the open ocean net pens produced transient increases in both plasma cortisol and glucose levels, suggesting a high level of stress associated with extended flounder handling and transfer.     

漠斑牙鲆仔鱼摄食节律的研究

在实验室条件下研究了漠斑牙鲆仔鱼摄食节律。试验结果表明：漠斑牙鲆仔鱼摄食具有昼夜节律性,其摄食活动主要在白天进行,在10：00光照度为400lx时摄食最为活跃,抵达摄食高峰,仔鱼夜间不摄食,全天表现明显的摄食节律。     

Fine tuning of feeding practices for hatchery produced Persian sturgeon,Acipenser persicus and Beluga sturgeon,Huso huso

In this work, we investigated the effects of various feeding treatments on the survival and growth of Huso huso and Acipenser persicus larvae during a 20‐day culture period. Three replicate groups (250 fish/replicate) of first‐feeding larvae were fed according to four main feeding regimes: (1) live food (live nauplii of brine shrimp Artemia urmiana); (2) indirect transition (5–7 days live food followed by gradual transition to formulated diet); (3) direct transition (using different combinations of live and formulated diet from the start feeding onwards); and (4) formulated feed (FD) from the start of feeding. In H. huso larvae, combining live food and manufactured diets (co‐feeding) from the first feeding stage onwards (direct transition) resulted in significantly higher weight gain than the other regimes. Survival was significantly higher in H. huso larvae fed solely live food or the direct transition regimes compared with indirect transition and FD. In A. persicus larvae, growth and survival were higher in the indirect transition feeding regime than in the other regimes. On the basis of the results of this study, we recommend co‐feeding of H. huso immediately from the commencement of exogenous feeding, but co‐feeding of A. persicus should start 7 days after prior feeding with live food.     

A Preliminary Study on the Effects of Salinity on Growth and Survival of Mulloway Argyrosomus japonicus Larvae and Juveniles

Abstract.— Tko experiments were conducted to determine the effects of salinity on growth and survival of mulloway Argyrosomus japonicus larvae and juveniles. First, 6-d-old larvae were stocked into different salinities (5, 12.5, 20, 27.5 and 35 ppt) for 14 d. Larvae grew at all salinities, but based on results for growth and survival, the optimum range of salinity for 6-d-old to 20-d-old larvae is 5–12.5 ppt. During this experiment larvae held in all experimental salinities were infested by a dinoflagellate ectoparasite, Amyloodinium sp. Degree of infestation was affected by salinity. There were very low infestation rates at 5 ppt (0.2 parasites/larva). Infestation increased with salinity to 20 ppt (33.1 parasites/larva), then declined with salinity to 35 ppt (1.5 parasites/larva). For the second experiment, juveniles (6.1 ± 0.1 g/fish) were stocked into different salinities (0.6, 5, 10, 20 and 35 ppt) for 28 d. Juveniles were removed from freshwater 3 d after transfer as they did not feed, several fish died and many fish had lost equilibrium. However, when transferred directly to 5 ppt. these stressed fish recovered and behaved normally. Trends in final mean weight and food conversion ratio of juvenile mulloway suggest that fish performed best at 5 ppt. Although salinity (5 to 35 ppt) had no significant ( P > 0.05) effect on growth, survival, or food conversion ratio of juveniles, statistical power of the experiment was low (0.22). Based on these results we recommend that mulloway larvae older than 6 d be cultured at 5 to 12.5 ppt. Optimum growth of juveniles may also be achieved at low salinities.     

The Effect of Temperature on First Feeding, Growth, and Survival of Larval Witch Flounder Glyptocephalus cynoglossus

Witch flounder Glyptocephalus cynoglossus has recently been identified as a candidate species for aquaculture in the northeastern United States and the Canadian Atlantic Provinces. This study investigated the optimal temperatures for witch flounder larval first feeding and for long term larval culture from hatching through metamorphosis. Maximum first feeding occurred between 15.0 and 16.2 C. Larvae did not survive beyond first feeding when reared at mean temperatures of 5.1, 10.4, or 19.5 C and were unable to initiate feeding at mean rearing temperatures below 6.0 C. At a rearing temperature of 15.0 C in 16-L tanks, mean larval survival to 60 days post hatch (dph) was 14.1%. Mean overall length-specific growth rate for larvae reared to 60 dph at 15.0 C was 3.5%/d and mean absolute growth was 0.62 mm/d. Subsequent larval growth at 15.6 C began to taper off towards 70 dph at the onset of weaning which overlapped with larval metamorphosis. Growth plateaued at 85 dph, followed by a rebound between 90 and 95 dph. Survival was 100% when weaning onto a dry, pelleted diet was initiated at 70 dph with a 10-d live diet co-feeding period. These results are favorable and encourage the further pursuit of commercial witch flounder culture.     

Larval growth, survival and development of Metapenaeus monoceros (Fabricius) cultured in different salinities

Larvae of Metapenaeus monoceros (Fabricius) at protozoea 1 (PZ1) stage were stocked in 2‐L glass flasks to investigate the effects of various salinities (25, 30, 35, 40, 45, 50 and 55 ppt) on growth and survival until the post‐larval (PL) stages. The PZ larvae were not able to tolerate a sudden salinity drop of over 10 ppt. Yet, an abrupt salinity increase of over 10 or even 15 ppt did not cause mortality. The PZ larvae were successfully acclimated to different test salinities at a rate of 4 ppt h^?1. The larvae displayed better tolerance to high rather than low salinities. The lowest and highest critical salinities appeared to be 22 and 55 ppt respectively. Taking into account survival, growth and development results, the optimal salinity for the larval culture of M. monoceros inhabiting the Eastern Mediterranean was 40 ppt. At this salinity, the PZ1 larvae were successfully cultured until PL1 stage within 11 days with 68% survival on a feeding regime of Tetraselmis chuii Kylin (Butcher) (20 cells μ L^?1), Chaetoceros calcitrans Paulsen (50 cells μ L^?1), Isochrysis galbana Parke (30 cells μL^?1) and five newly hatched Artemia nauplii mL^?1 from M1 onwards at 28 °C.     

Effects of dietary docosahexaenoic acid (22:6n‐3) and arachidonic acid (20:4n‐6) on the growth,survival, stress resistance and fatty acid composition in black sea bass Centropristis striata (Linnaeus 1758) larvae

The objectives of this study were to determine the effects of the dietary docosahexaenoic acid (DHA) to arachidonic acid (ARA) ratio on the survival, growth, hypersaline stress resistance and tissue composition of black sea bass larvae raised from first feeding to metamorphic stages. Larvae were fed enriched rotifers Brachionus rotundiformis and Artemia nauplii containing two levels of DHA (0% and 10% total fatty acids=TFA) in conjunction with three levels of ARA (0%, 3% and 6% TFA). On d24ph, larvae fed the 10:6 (DHA:ARA) treatment showed significantly (P<0.05) higher survival (62.3%) than larvae fed 0:0 (DHA:ARA) (27.4%). Notochord length and dry weight were also significantly (P<0.05) greater in the 10:6 (DHA:ARA) treatment (8.65 mm, 2.14 mg) than in the 0:0 (DHA:ARA) (7.7 mm, 1.65 mg) treatment. During hypersaline (65 g L⁻¹) challenge, no significant differences (P>0.05) were observed in the median survival time (ST50) between larvae fed 10% DHA (ST50=25.6 min) and larvae fed 0% DHA (ST50=18.2 min). The results suggested that black sea bass larvae fed prey containing 10% DHA with increasing ARA within the range of 0–6% showed improved growth and survival from first feeding through metamorphic stages.     

Sustained, Natural Spawning of Southern Flounder Paralichthys lethostigma Under an Extended Photothermal Regime

Hormone‐induced spawning of southern flounder Paralichthys lethostigma has produced substantial numbers of viable eggs, but wide variations in fertilization and hatch rates have been reported. Recently, sustained natural spawning of southern flounder broodstock, without hormone induction, has been achieved in our laboratory. Adults (average weight = 1.12 kg; N= 25), including 6 captured as juveniles in 1993 and 19 captured as adults during September 1998, were stocked in two 4.8‐m³ controlled‐environment tanks in October 1998 and held under natural photothermal conditions until January 1999, when an artificial winter photo‐period of 10 L:14 D was initiated and then maintained through April 1999. Sex ratio was approximately 13 females:8 males:7 unknown. Natural spawning was observed in early December 1998 and increased in frequency to a peak in March 1999, before declining in late April. Water temperature ranged from 13.9 to 24.5 C during the spawning period. Natural spawnings over 142 d produced a total of 18.3 × 10⁶ eggs, with a mean fertilization rate of 28.0% (range = 0–100%), yielding 4.94 × 10⁶ fertilized eggs. The mean percentage of eggs that remained buoyant in full‐strength seawater (34 ppt) was 41.3% (0–98%), while hatching rate of buoyant eggs was 37.3% (0–99%) and survival of yolksac larvae to the first‐feeding stage was 30.2% (0–100%). Gonadal biopsies in late April identified six females from both tanks as probable spawners. A preliminary comparison suggests that natural spawning produced much larger numbers of viable eggs per female, with higher egg quality (i.e., fertilization and hatching success) than hormone‐induced spawning. In contrast to natural spawning, hormone‐induced strip‐spawning enabled timing of spawnings to be more precisely controlled. These results suggest that a combination of both natural and hormone‐induced spawning of photothermally conditioned fish will help produce the large numbers of eggs required to support commercial production.     

Development of a Squid-hydrolysate-based Larval Diet and its Feeding Performance on Summer Flounder, Paralichthys dentatus, Larvae

Locally generated squid‐processing byproduct was processed into concentrated hydrolysate (22% solids, 17.3% protein, and 3.0% lipid, primarily phospholipids—11.6% eicosapentaenoic acid/24.5% docosahexaenoic acid on a lipid weight basis). Two microparticulate diets (65% protein, 19% lipid, 7.5% carbohydrate, and 19.12 MJ/kg energy, on a dry weight basis) were prepared using squid hydrolysate (SH) and squid‐herring hydrolysate as sole protein sources (73.3 and 78.65% of the whole diet, respectively). A 22‐d feeding trial with summer flounder, Paralichthys dentatus, larvae of 17 d after hatch showed that the survival rate (92%) of larvae fed SH was significantly (P < 0.05) higher than those of larvae fed live Artemia nauplii (81%) and a commercial diet, Proton (65%), while specific growth rates (SGR) were comparable (2.23% /d for SH and 2.86% /d for Artemia) with the lowest for Proton (1.39% /d). After switching from commercial and Artemia diets to a SH diet for 17 d following the 22‐d feeding, significant improvements were seen in survival rates of postweaning larvae fed previously commercial (65.28–76.57%) and Artemia diets (81.25–89.07%).