Optimal Stocking Density for Juvenile Winter Flounder Pseudopleuronectes americanus

Abstract.— A 12-wk experiment was conducted to test the effects of 50, 100, 200, and 300% stocking densities (ventral fish area to bottom tank area ratio) on growth, survival, and aggressive behavior in 2-moold juvenile winter flounder Pseudopleuronectes americanus . Over the course of the experiment, there were no differences ( P > 0.05) in growth between treatments at any given week. Survival decreased in all treatments over time, however, proportionately more fish died in the high (200 and 300%) density treatments. At the end of 12 wk, the 300% treatment had significantly lower survival ( P < 0.05). There were no differences in the degree of caudal fin damage between any of the treatments ( P > 0.05) suggesting that aggressive behavior. such as fin nipping, was not affected by stocking density. However, fish size was inversely related to caudal fin damage. When fish were sorted by length, the two smallest size categories (<2 cm and 2-3 cm) suffered the greatest fin nipping compared to each other ( P < 0.001) and all larger size categories ( P < 0.001). These results indicate that 50 to 300% stocking densities do not affect growth or aggressive behavior in juvenile winter flounder. Although survival was affected by density, the final yield (weight per treatment) at higher densities was still greater than at lower densities. Our hatchery recommendations are to stock at ≥200% but to monitor fish health carefully. Size grading should be employed to minimize aggression between fish size classes.     

Gonadal Development and Differentiation in Cultured Juvenile Winter Flounder, Pseudopleuronectes americanus

Winter flounder, Pseudopleuronectes americanus, is currently being evaluated as a stock enhancement candidate in New Hampshire, USA; however, little is known about the gonadal development or the sex ratio of cultured juveniles. To determine the size at gonadal differentiation, 327 cultured fish ranging from <20 to 110 mm total length (TL), in 10‐mm‐TL size classes, were examined histologically. Gonads had differentiated into testes and ovaries in fish ≥41 mm TL (98%), whereas the majority of fish (81%) smaller than 40 mm TL possessed undifferentiated gonads. A total of 313 cultured fish >40 mm TL were analyzed for sex ratio. In 2003, 67 females and 164 males were identified, yielding a sex ratio that was significantly skewed toward male (χ²= 40.7, df = 1, P < 0.001). This trend held true when cultured fish were sorted by age and length, with the exception of those fish 61–70 mm TL. This aberration probably was because of a small sample size in this length category. However, in both the 2004 and the 2005 cultured populations, flounder sex did not deviate from a 1:1 ratio (2004 χ²= 0.12, df = 1, P= 0.724 and 2005 χ²= 0.02, df = 1, P= 0.881). The 2003 data suggest that environmental or genetic factors may affect winter flounder sex determination; rearing manipulation studies in the hatchery are needed to confirm this hypothesis.     

The Effects of Live and Artificial Diets on Feeding Performance of Cultured Winter Flounder,Pseudopleuronectes americanus,in the Wild: Survival,Feeding, Growth,and Nucleic Acid Analyses

This research, which is part of a larger study designed to assess the feasibility of winter flounder, Pseudopleuronectes americanus, stock enhancement in New Hampshire, identifies hatchery feeds that optimize feeding‐related performance of fish once released in the wild. Fish reared on post‐nauplii of brine shrimp, Artemia sp., white worms, Enchytraeus albidus, common burrower amphipods, Leptocheirus plumulosus, and formulated pellets were evaluated post‐release from in situ cages using survival, growth rate, feeding onset and incidence, stomach fullness, diet composition, and nucleic acid‐based condition as indicators of hatchery diet suitability. Amphipod‐reared fish had the highest mean stomach content index of all feed types, including wild fish. Wild and worm‐reared fish exhibited the most similar survival, overall stomach fullness, and diet composition profiles over time. Amphipod‐reared fish ranked highest in overall performance; however, if wild fish performance is viewed as the ideal for a stocked fish, worm‐reared fish performed optimally. This study describes hatchery feeding strategies that may ease the transition of flatfish released into the wild for stock enhancement.     

The Effect of Detergents on Solubilization and Activity of the Lipid Peroxidative Enzyme System from the Sarcoplasmic Reticulum of Winter Flounder (Pseudopleuronectes americanus)

Sarcoplasmic reticular (SR) membranes from flounder muscle were solubilized using different ionic and nonionic detergents (Tween-20, Triton X-100, sodium dodecyl sulfate, or sodium desoxycholate). The recoveries of protein and NADH-dependent lipid peroxidative enzyme activity for different detergent treated SR preparations were compared. Triton X-100 was the least detrimental to the activity of the solubilized enzymes system. With Triton X-100, we were able to solubilize 85% of the SR protein and 97% of the phospholipid. Upon removal of the detergent, 98% of the peroxidative activity was recovered.     

不同摄食水平对牙鲆幼鱼生长及能量收支的影响

在水温(20.0±0.5)℃,摄食水平分别为体质量的1.0%、1.5%及饱食条件下,进行牙鲆幼鱼[体质量为(35.08±0.05)g]的生长和能量收支的研究。试验结果表明,随着摄食水平的增加,鱼体干物质、蛋白质和能量表观消化率随摄食水平的增加而增加;湿质量特定生长率随摄食水平的增加呈线性上升,干物质、蛋白质和能量特定生长率呈对数增加;饲料湿质量转化效率不受摄食水平的影响,饲料干物质、蛋白质和能量转化效率随摄食水平的增加先升后降。粪能占摄食能的比例随摄食水平的增加而显著降低,生长能占摄食能的比例随摄食水平的增加先升后降,代谢能占摄食能的比例变化与生长能占摄食能的比例变化相反。各个摄食水平下能量收支式为:1.0%:100.00C=28.99G+2.34F+6.03U+62.64R;1.5%:100.00C=37.19G+1.94F+5.27U+55.60R;饱食:100.00C=30.73G+1.36F+5.16U+62.75R。     

Effects of Dietary Protein and Energy Levels on Growth and Body Composition of Juvenile Flounder Paralichthys olivaceus

A feeding trial of three protein levels (30, 40 and 50%) and two energy levels (300 and 400 kcal/100-g diet) factorial design with three replications was carried out to investigate the proper dietary protein and energy levels for the growth of juvenile flounder Paralichthys olivaceus . Weight gain of fish tended to improve with increasing dietary protein level. Weight gain of fish fed either the 40% or 50% protein diet with 300 kcal/100-g diet was significantly higher ( P < 0.05) than with 400 kcal/100-g diet. The best weight gain was obtained from fish fed the 50% protein diet with 300 kcal/100-g diet. Feed efficiency tended to improve with increasing dietary protein level. However, dietary energy level had no significant effect on feed efficiency of fish fed the 30% or 50% protein diet, but that of fish fed the 40% protein diet with 300 kcal/100-g diet was significantly higher than with 400 kcal/100-g diet. Protein retention tended to increase as dietary protein level increased and energy level decreased. Lipid content of fish fed the diet containing 400 kcal/100-g diet was significantly higher than that of fish fed the diet containing 300 kcal/100-g diet at all protein levels. Fatty acid compositions such as linoleic acid, EPA (20:5n-3) and DHA (22:6n-3) offish were directly affected by dietary lipid (squid liver oil and/or soybean oil) used for energy source. Based on the above results, it can be concluded that the proper dietary protein and energy levels for the growth of juvenile flounder are 50% and 300 kcal/100-g diet, respectively.     

Weaning of Wild Young-of-the-Year Winter Flounder Pleuronectes americanus (Walbaum) on a Dry Diet: Effects on Growth, Survival, and Feed Efficiency Ratios

Winter flounder Pleuronectes americanus is being developed for aquaculture in Atlantic Canada. Weaning, the switch from a live diet to formulated feed, is a critical stage in the culture of any fish species. In this study, we used wild young-of-the-year stock to determine whether juvenile winter flounder can be weaned from live, cultured Artemia onto dry feed. The dry feed used was originally formulated for turbot Scophthalmus maximus and cod Gadus morhua by BP Nutrition, Stavanger, Norway. The performance of live feed and dry pellets was compared in terms of survival, growth and feed efficiency over three months. We were able to wean wild juvenile winter flounder onto dry feed after one week. Increased specific growth rates and feed efficiency ratios in weaned juveniles indicated that prepared diet was a better food source than Anemia . Survival was not influenced by food type.     

盐度对漠斑牙鲆幼鱼存活、生长及摄食的影响

盐度对漠斑牙鲆幼鱼的存活的影响较小,将漠斑牙鲆幼鱼从盐度为32的海水直接放入盐度为24、16、8、0的水中观察48 h存活情况,仅盐度0组成活率为75%,其他盐度组均为100%.在盐度为32、24、16、8、0各试验组中,经1个月饲养,生长速度和饵料系数均无显著差异.盐度越低越有利于漠斑牙鲆白化个体康复正常体色.     

Effects of Water Velocity on Growth Performance of Juvenile Japanese Flounder Paralichthys olivaceus

Critical swimming speed (Ucrit) of juvenile Japanese flounder Paralichthys olivaceus with 12.5-cm body length was determined to be 1.8-body lengthkec. A feeding trial was conducted to investigate the effects of water velocity on growth and other nutritional parameters of juvenile flounder (initial mean body weight: 5.7 g/fish), which is a sedentary species. The fish were raised in three water velocities, < 0.3 body length/sec (control), 0.9 body length/sec (slow) and 2.1 body length/sec (moderate), for 8 wk. Over the course of the feeding trial, the weight gain of the control, slow and moderate groups of fish were. 634%. 671% and 564%. respectively (significant between slow and moderate groups). A second order polynomial suggested that the optimum water velocity occurred at about 1.0 body length/sec. The feed efficiency of the moderate group (1.48) was significantly lower than those of the control (1.56) and slow (1.56) groups, while there were no significant differences in feed intake (1.72–1.75% of wet biomass) among the treatments. Water velocity did not affect the proximate composition of the whole body; however, rearing the fish at the moderate water velocity did slightly but significantly reduce protein retention. Lipid content of fin muscles was significantly different among the treatments: control (14.9%), slow (17.6%) and moderate groups (11.6%). Unlike salmonids, water current does not seem effective for improving growth and feed efficiency in juvenile Japanese flounder.     

The Effect of Different Dietary Energy Levels on Growth and Survival of Eyestalk Ablated and Intact Juvenile Lobsters Homarus americanus

When fed formulated diets containing 45% protein derived from the rock crab Cancer irroratus , wheat gluten, gelatin, lipids adjusted from 3–14% and carbohydrates adjusted from 5–24%, the average survival of bilaterally eyestalk ablated juvenile lobsters and intact controls were 91% and 99%, respectively over a 90-day experiment. Photopenad (13 light: 11 dark vs 3 light: 21 dark) did not have a significant effect on growth or survival of ablated or intact lobsters. The optimal digestible energy levels and protein/energy ratio for the greatest growth in both ablated and intact lobsters were 3.7 to 3.8 (kcal/g dry) and 0.12 [g protein/(kcal/g)], respectively. The best feed conversion ratios (0.97 and 1.40 diet given/weight gain) for ablated and intact lobsters were obtained with diets containing 3.83 kcal/g and 3.63 kcal/g diet, respectively. The present study also demonstrated that crab protein served as a good protein source for intact and ablated lobsters.     

Secondary Stress Responses in Juvenile Brazilian Flounder,Paralichthys orbignyanus,throughout and after Exposure to Sublethal Levels of Ammonia and Nitrite

This study investigated the secondary stress responses of Paralichthys orbignyanus exposed to ammonia and nitrite and after recovery. Fish were exposed to 0.12, 0.28, and 0.57 mg NH₃‐N/L, or 5.72, 10.43, and 15.27 mg NO₂‐N/L for 10 d followed by the same time length for recovery. Ammonia‐ and nitrite‐free water was used as a control treatment. Blood samples were collected after 1, 5, and 10 d of exposure and after recovery. Fish exposed to ammonia presented lower and higher glucose levels after 10 d of exposure and recovery, respectively. Ammonia induced initial and transient ionic disturbances and metabolic alkalosis. Nitrite exposure caused hyperglycemia, increased plasma K⁺ levels, and respiratory alkalosis, whereas metabolic acidosis was observed after recovery. Increased proportion of monocytes and/or granulocytes and reduced number of lymphocytes were demonstrated in fish exposed to 0.28 mg NH₃‐N/L (Day 1) and 10.43 mg NO₂‐N/L (Day 5) and after recovery in the 0.28 and 0.57 mg NH₃‐N/L treatments. Exposure to ammonia decreased the proportion of granulocytes on Day 5. In conclusion, exposure to concentrations at 0.12 mg NH₃‐N/L and 5.72 mg NO₂‐N/L provoked physiological disorders in Brazilian flounder. Nonetheless, fish exposed to 5.72 mg NO₂‐N/L following a 10‐d recovery period showed complete resumption of homeostasis.     

Effects of Dietary Protein and Lipid Levels on Growth and Body Composition of Juvenile Japanese Flounder,Paralichthys olivaceus,Fed Diets Containing Intact Proteins or Crystalline Amino Acids

Abstract

The influence of eight semi-purified diets having different protein and lipid levels on growth performance and body composition of Japanese flounder, Paralichthys olivaceus, was evaluated. The sources of protein were either intact protein (IP) or a mixture of IP and crystalline amino acids (CAA). In trial 1 (IP group), four test diets were prepared from IP sources (casein-gelatin) with combination of two protein levels (35% and 50%) and two lipid levels (10% and 20%). In trial 2 (PA-group), another four test diets were formulated using IP and CAA with combination of the same two protein and two lipid levels as used for trial 1. The juveniles (about 2.00 g initial weight) were fed the respective test diets for 42 days in duplicate tanks. In both trials, weight gain was the highest for juveniles fed the diet containing 50% protein with 10% lipid and the lowest in those fed the 35% protein with 10% lipid diet. Weight gain of juveniles was significantly increased when the lipid levels were increased in low protein (35%) diets for both trials. In high protein (50%) diet, however, increase lipid levels from 10 to 20% with 50% protein resulted in decrease in weight gain, irrespective of protein sources. Specific growth rate, feed intake, feed conversion efficiency were significantly affected by different dietary protein and lipid level. Content of neutral lipids in the whole-body particularly, triglycerides was statistically higher for fish fed the high-lipid diets than those fed the low-lipid diets irrespective of protein sources. But, polar lipid did not show any differences either for different protein or lipid levels. Level of individual amino acids in the whole-body showed little difference among the dietary treatments. Results suggest that Japanese flounder grew better on low lipid level with 50% protein-based diets and supplementation of CAA did not show any adverse effect on the growth performance with different levels of protein and lipid in the diets.     

Effects of Dietary Chlorella ellipsoidea Supplementation on Growth, Blood Characteristics, and Whole-Body Composition in Juvenile Japanese Flounder Paralichthys olivaceus

The study was conducted to investigate the effects of dietary Chlorella powder (C) supplementation on growth performance, blood characteristics, and whole-body composition in juvenile Japanese flounder Paralichthys olivaceus . Four experimental diets were supplemented with C at 0, 1, 2 and 4% (C₀, C₁, C₂, and C₄) on a dry-weight basis. Three replicate groups of fish averaging 1.13 ± 0.02 g (Mean ± SD) were randomly distributed in each aquarium and fed one of four experimental diets for 12 wk. After 12 wk of the feeding trial, fish fed C₂ diet had higher weight gain (WG), feed efficiency (FE), and protein efficiency ratio (PER) than did fish fed C₀ and C₁ diets ( P 0.05); however, there was no significant difference among fish fed C₁ and C₄ diets, and among fish fed C₂ and C₄ diets ( P > 0.05). Fish fed C₂ and C₄ diets had a lower serum cholesterol level than did fish fed C₀ and C₁ diets ( P 0.05). Fish fed C₂ and C₄ diets had a lower body fat than did fish fed C₀ ( P 0.05). These results indicate that dietary supplementation of 2% Chlorella powder in the commercial diets could improve growth, feed utilization, serum cholesterol level, and whole-body fat contents in juvenile Japanese flounder.     

Effects of Putative Growth or Health‐Enhancing Dietary Additives on Juvenile Olive Flounder,Paralichthys olivaceus,Performance

The effect on growth and body composition of various dietary additives with putative growth or health‐enhancing properties were determined in juvenile olive flounder (25 g initial weight). Nine experimental diets were prepared to contain one of the following additives: control (Con) with no additive, Opuntia ficus‐indica ver. saboten (OF), propolis (PP), lactic acid bacteria (LA), γ‐poly‐glutamic acid (PG), onion extract (OE), organic sulfur (OS), Biostone® (BS), and fig extract (FE). Fishmeal, dehulled soybean meal, and corn gluten were used as the protein source of the experimental diets. Wheat flour and soybean oil were used as the carbohydrate and lipid sources, respectively. Dietary additives were included in each experimental diet at 1% at the expense of wheat flour except for the FE (aqueous), which was substituted at 1% of the amount of water added to the diet. Fish were hand‐fed to satiation twice a day for 6 d/wk for 6 wk. Weight gain of fish fed the OE diet was higher than that of fish fed with the PP diet. Chemical composition of fish was not different among the experimental diets. OE was the most effective dietary additive to improve performance of olive flounder among additives used in this study.     

摄食豆肽对星斑川鲽(Platichthys stellatus)幼鱼肝脏蛋白表达的影响

为揭示饲料中添加豆肽对星斑川鲽(Platichthys stellatus)幼鱼肝脏代谢产生的影响,利用大豆蛋白肽替代24%的鱼粉并以全鱼粉饲料为对照形成两种等氮等能的实验饲料,粗蛋白为(43.98±0.14)%、能量为(19.89±0.08) kJ/g。将240尾幼鱼分为两组,每组3个平行(40尾/桶),分别投喂两种饲料50 d后,提取肝脏蛋白进行双向电泳并用PDQuest分析差异蛋白。结果显示,在pH 4?7、分子量14.4?97.4 kDa范围内,共检测到448?470个肝脏蛋白点,有14个蛋白的表达呈两倍以上的差异,其中3个为结构蛋白(中间纤维蛋白、II型细胞骨架8-样异构体X2、肌动蛋白),其他11种肝脏蛋白参与不同代谢活动。以摄食全鱼粉饲料的肝脏蛋白为对照,幼鱼摄食豆肽饲料后明显上调了9种代谢蛋白和3种结构蛋白,下调了两种代谢蛋白。本研究结果表明,星斑川鲽幼鱼摄食豆肽饲料后其肝脏代谢发生变化,即提高高密度脂蛋白的生成,增强糖酵解、降低糖原合成从而使代谢向产能方向进行,降低胰蛋白酶的合成和提高应激蛋白的生成。本研究利用比较蛋白组学技术从肝脏蛋白表达的角度来揭示摄食豆肽对星斑川鲽幼鱼肝脏组织代谢的影响,为新型植物蛋白源的应用提供理论基础。     

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.     

Effects of Larval Stocking Density on Laboratory-Scale and Commercial-Scale Production of Summer Flounder Paraliehthys dentatus

Abstract.— Three experiments investigating larval stocking densities of summer flounder from hatch to metamorphosis, Paralichthvs dentalus, were conducted at laboratory‐scale (75‐L aquaria) and at commercial scale (1,000‐L tanks). Experiments 1 and 2 at commercial scale tested the densities of 10 and 60 larvae/L, and 10, 20, and 30/L, respectively. The laboratory scale experiment tested the densities of 10, 20, 30, and 40 larvae/L. Experiments were carried out in two separate filtered, flow‐through seawater systems at URI Narragansett Bay Campus (laboratory‐scale), and at GreatBay Aquafarms, Inc. (commercial‐scale). At both locations, the larvae were raised in a “greenwater” culture environment, and fed rotifers and brine shrimp nauplii according to feeding regimes established for each location. Water temperature was maintained at 21C (± 2) and 19C (± 1) for the duration of laboratory and commercial experiments, respectively. Experiments 1 and 2 at the commercial location were terminated at 42 and 37 d post hatch (dph), respectively, and the laboratory experiment lasted 34 DPH. Larvae initially stocked at 10/L grew to an average length of 14.3 and 14.4 mm, and were significantly larger (P < 0.05) than those stocked at 30/L (13.1 mm) and 60/L (11.7 mm) in commercial scale experiments I and 2, respectively. At laboratory scale, no significant differences in length were detected, although mean total length tended to decrease with increasing stocking density (average length of 14.2, 13.3, 12.7, and 12.7 mm for treatments of 10, 20, 30, and 40/L, respectively). Final survival percentage was not affected by stocking density in either commercial experiment, and was 61 and 40% for treatments of 10 and 60/L in Experiment 1, respectively, and 62, 59, and 56% for Experiment 2, respectively. Similarly, there was no significant difference in final survival percentage among treatments in the laboratory experiment, which averaged 59, 55, 56, and 37% for treatments of 10, 20, 30, and 40L. respectively. Since larval length was not different between the intermediate densities (20 and 30 Iarvae/L), and because high‐density rearing can produce a much greater numerical yield per tank, we recommend a density of 30 larvaen as an optimal stocking density for the hatchery production of summer flounder.     

微生态制剂对牙鲆幼鱼脂肪酶的影响

试验结果表明,微生态制剂可提高牙鲆幼鱼消化道的脂肪酶活性,微生态制剂的使用方式和使用的间隔时间影响其使用效果。拌饲投喂与在水中同时使用微生态制剂,在7 d内使脂肪酶活性大大提高,但随后采用这种使用方式的试验组脂肪酶活性又迅速降低。而只在水中使用微生态制剂的各试验组的脂肪酶活性却随着试验的进行,逐步提高。有利于脂肪酶活性提高的最佳方案是只在水中使用微生态制剂,使用间隔时间为7 d,芽孢杆菌∶乳酸菌为10∶1,菌液密度为105/m l。