Characterization of chymotrypsin activity during early ontogeny of larval red drum (Sciaenops ocellatus)

The temporal evolution of chymotrypsin activity during early ontogeny of laboratory reared red drum larvae was accomplished using a combination of biochemical assays and electrophoretic methods (substrate SDS-PAGE). Optimal functional conditions for chymotrypsin were also determined. Chymotrypsin activity was first detected prior to the onset of exogenous feeding. Total chymotrypsin activity increased with age and standard length. Specific activity was greatest on day 10 post-hatch. Maximal chymotrypsin activity was observed at 50 °C, pH 7.8, and Ca²⁺ concentration of 25 mM. Using substrate gel electrophoresis and specific inhibitors the molecular weight of red drum chymotrypsin was estimated to be 26–27 kD. Our results indicate that the digestive system of red drum larvae is capable of alkaline proteolysis before first feeding and suggest that chymotrypsin may have potential as an indicator of nutritional condition.     

美国红鱼形态和生长参数的研究

用传统法和框架法（Truss) 测定了红鱼形态的可数可量怀状，分析了红鱼形态有关参数的相互关系，拟合了红鱼的框架结构图。总结回归了不同养殖环境中红鱼的体长，体重之间的关系。比较分析了不同养殖条件下红鱼的日生长，生长率等指标，不同群体的生长特征指数生长拐点和生长方程。本研究为选育红鱼良种和检测原，良种提供了科学依据。     

美国红鱼的营养成分分析与评价

测定了养殖红鱼肌肉及鱼体中的蛋白质、脂类、灰分等生化组成，并计算了其比能值，分析了肌肉蛋白质中17种常见氨基酸的含量。结果表明，红鱼肌肉中脂类含量低，属高蛋白低脂肪、低比能值鱼类，肌肉氨基酸含量同其他经济鱼类相比，属中等水平。必需氨基酸的含量低且缺含硫氨基酸，为非完全蛋白。其呈味氨基酸百分含量较其他鱼类高。     

饲料维生素E含量对美国红鱼生长和非特异性免疫的影响

采用单因子实验设计方法,进行了饲料中添加维生素E对美国红鱼生长（存活率、特定生长率和饲料效率）和非特异性免疫相关指标（血清中溶菌酶活性、血清总补体活性和超氧化物岐化酶（SOD）活性）影响的研究,设计了5个不同维生素E水平（0．1、25．3、51．0、102．8和203．2mg／kg）的等氮等能饲料,对美国红鱼进行为期56d的生长实验,每个水平3个重复,每个养殖单元放养初始体重为151g左右的美国红鱼20尾。饲养实验在海水网箱（1．5m×1．0m×1．0m）中进行。结果表明,随着饲料维生素E含量增加,美国红鱼饲料效率、特定生长率和成活率显著提高,美国红鱼最佳生长性能的饲料维生素E添加量为102．8mg／kg;美国红鱼血清溶菌酶和血清总补体活性随着维生素E的添加量的增加而显著升高,并且维生素E的添加量达到最高值102．8和25．3mg／kg,随后变化不明显;美国红鱼血清SOD活性随维生素E添加量的增加而显著升高,并且含量在102．8mg／kg达到最高值,随后显著下降。综上所述,美国红鱼维生素E的适宜添加量为102．8mg／kg。     

逃逸美国红鱼在浙江海域生长特性的初步研究

根据2004~2006年采集的浙江海域156尾美国红鱼的生物学测定资料,分析了该鱼种可量性状间关系,回归得出全长-体长、体重-纯重、体长-体重关系,并比较分析了其他研究者的结果。采用美国红鱼矢耳石磨片法鉴定年龄,并用FiSATⅡ软件拟合得出von-Bertalanffy生长方程,其中:BL∞=951·09mm,k=0·27,t0=0·57;TL∞=1144·61mm,k=0·26,t0=0·52;WB∞=14468·07g,结合生长拐点(tx=4·514)、生长特征指数(Φ′BL=5·388)及肥满度指标(Q=1·82)。比较分析不同生态环境下美国红鱼的生长情况,表明浙江海域美国红鱼生长特性最接近于美国自然海区,浙江海域适合其生长,一旦形成种群可能会对本地种造成威胁。     

Dietary vitamin C requirement of red drum Sciaenops ocellatus

A feeding trial was conducted to determine the minimum dietary vitamin C requirement of juvenile red drum and characterize signs of vitamin C deficiency. Semipurified diets containing 400 g crude protein kg^–1 from lyophilized red drum muscle and crystalline amino acids were used in the feeding trial. The basal diet without supplemental vitamin C contained ≈ 6 mg vitamin C kg^–1 and was fed for a 1-week conditioning period. Red drum, initially averaging ≈ 3.6 g each, were fed diets supplemented with 0, 10, 20, 30, 45, 60, 75, or 150 mg vitamin C kg^–1 as ascorbate polyphosphate for a period of 10 weeks. Fish fed the basal diet began to exhibit overt signs of vitamin C deficiency, including reduced weight gain, lordosis, scoliosis and loss of equilibrium after 8 weeks. Total ascorbate was undetectable in liver of fish fed the basal diet while liver ascorbate levels in fish fed the other diets generally reflected dietary supplementation. Regression analysis of weight gain data using the broken-line model resulted in a minimum vitamin C requirement ( ± SE) of 15 ± 3 mg kg^–1 diet.     

Optimization of growout diets for red drum,Sciaenops ocellatus

Because of the high costs associated with feed inputs, as well as increased concern about waste production on fish farms, there is considerable interest in developing growout diets which are both cost effective and low polluting. In two 12‐week growth trials, the response of subadult red drum, Sciaenops ocellatus, fed either a diet of 440 or 360 g protein kg^?1 diet (44% or 36%) with varying E:P ratios were tested. In the first experiment, five diets containing 440 g protein kg^?1 diet and one diet containing 360 g protein kg^?1 diet (reference) were offered to red drum (mean initial weight of 186 g). The five test diets contained 83, 103, 123, 143, and 163 g lipid kg^?1 diet, resulting in E:P ratios ranging from 34.3 to 38.9 kJ g protein^–1. In experiment 2, five diets providing 360 g protein kg^?1 diet and one diet containing 440 g protein kg^?1 diet (reference) were offered to red drum (mean initial weight of 145 g). Dietary lipid levels included 83, 123, and 163 g lipid kg^?1 diet, and dietary carbohydrate was diluted with 10% and 20% non‐nutritive bulk filler in two of the diets to result in E:P ratios ranging from 34.5 to 46.7 kJ g protein^–1. In experiment 1, no significant differences in mean final weight, mean weight gain, feed efficiency, protein conversion efficiency or hepatosomatic index were observed between the five test diets providing 440 g protein kg^?1 diet. Intraperitoneal fat generally increased with increasing dietary lipid. The results of experiment 2 indicate that amongst the test diets with 360 g protein kg^?1 diet, mean final weight, mean weight gain, feed efficiency, protein conversion efficiency and hepatosomatic index were not significantly different. Intraperitoneal fat significantly increased with increasing dietary lipid. In both experiments, fish offered diets with 440 g protein kg^?1 diet produced significantly higher growth and FE values as compared to fish receiving diets containing 360 g protein kg^?1 diet. This study indicated that subadult red drum are tolerant of shifts in E:P ratios and utilize a wide range of dietary lipid and carbohydrate without compromising growth.     

Tests of Mendelian segregation and linkage-group relationships among 31 microsatellite loci in red drum, Sciaenops ocellatus

Two families (parents and full-sibling progeny) of red drum (Sciaenops ocellatus) were assayed for genotypes at 31 nuclear-encoded microsatellites. Tests of expected Mendelian segregation at 29 of the microsatellites were non-significant following Bonferroni correction; occurrence of null alleles was inferred at the remaining two microsatellites. Nine of the 24 possible linkage groups (range 2–5 microsatellites per linkage group) were identified. Significant differences in recombination rates were found both between sexes and between families. The microsatellite linkage groups generated in this study represent the beginning of a genetic map for red drum, an economically important marine fish in the southeastern region of the United States of America.     

Dietary vitamin E requirement of the red drum Sciaenops ocellatus

A 12-week feeding trial was conducted to establish the minimum dietary vitamin E requirement of juvenile red drum by broken-line regression analysis. The semi-purified basal diet was supplemented with 10, 20, 30, 40, 60 or 80 IU vitamin E kg⁻¹ as all-rac -α-tocopheryl acetate. Juvenile red drum were conditioned by feeding the basal diet for 8 weeks prior to the feeding trial to reduce whole-body vitamin E levels. Then, fish initially averaging 12.2 ± 0.4 g fish⁻¹ (mean ± SD) were fed the experimental diets at a rate approaching apparent satiation for 12 weeks. Weight gain and feed efficiency responses of fish fed diets were significantly ( P  < 0.01) altered by the level of vitamin E supplementation but not strictly in a dose-dependent manner. Vitamin E concentrations in liver and plasma also were significantly ( P  < 0.001) influenced by dietary vitamin E level. Plasma ascorbic acid in fish fed the basal diet tended ( P  = 0.066) to be lower than in fish fed diets containing the various levels of vitamin E. In addition, fish fed the basal diet showed edema in the heart, while fish fed all other diets were normal. Fish fed 60 or 80 IU all-rac -α-tocopheryl acetate kg⁻¹ diet had significantly higher respiratory burst of head kidney macrophages than fish fed all other diets, although dietary effects on hematocrit and neutrophil oxidative radical production were not significant. The minimum dietary vitamin E requirement of juvenile red drum was established based on broken-line regression of liver thiobarbituric acid reactive substances to be 31 mg all-rac -α-tocopheryl acetate kg⁻¹ diet.     

Heritability of juvenile growth traits in red drum (Sciaenops ocellatus L.)

Heritability of juvenile growth rate was estimated for red drum (Sciaenops ocellatus), an economically important sciaenid fish in the southern USA. Thirty‐eight families were generated via ‘natural’ spawning of multiple sets of five breeders (three dams × two sires) in individual brood tanks. Offspring were individually tagged with Passive Integrated Transponder (PIT) tags and mixed for grow‐out in replicate ‘common‐garden’ tanks. Juvenile growth was followed from 166.4±18.6 to 254.0±27.0 mm (total length). Offspring were assigned a posteriori to individual brood fish (dam and sire) based on genotypes at nuclear‐encoded microsatellites. Heritability (h²) of a thermal growth coefficient was estimated using an animal‐additive model and a restricted maximum‐likelihood algorithm. Estimates of h² were 0.33±0.08 and 0.31±0.08 for thermal growth coefficient based on length and weight respectively. These results indicate a significant genetic component in juvenile growth rate in red drum. Estimates of h² for condition coefficient (K) at various measurement dates averaged 0.38, suggesting a genetic component to shape in juvenile red drum.     

Dietary effects on thyroid hormones in the red drum,Sciaenops ocellatus

Juvenile red drum (Sciaenops ocellatus) were cultured at 25°C on a variety of diets and blood sampled over eight weeks to examine the relationship between growth and plasma thyroid hormone levels. Maximum growth rates were achieved on formulated experimental diets and a simulated natural shrimp diet. Associated with these maximal rates was a significant increase in triidothyronine (T₃), but no consistent change in thyroxine (T₄). Reduced rations of diets resulted in low growth rates associated with significantly lowered levels of T₃ but not T₄. To determine whether weight gain could be increased by application of exogeneous hormone, diets were supplemented with T₃ or T₄ at 2, 10, and 50 mg hormone/kg diet. Significantly elevated T₃ was induced by supplementation with 10 and 50 mg T₃/kg diet, although there were no indications of an anabolic effect of T₃ incorporation, and 50 mg T₃/kg diet was in fact associated with decreased weight gain. Incorporation of T₄ into diets had no effect on growth or T₃, and had effects on T₄ which were small and inconsistent, indicating that T₄ may not be effectively absorbed from the gut. No difference was found in response to hormone feeding between low (6 ppt) or high (35 ppt) water salinity. T₃ levels thus appear to closely parallel growth in fish on unsupplemented diets, whereas T₄ which were small and manipulation. Supplementation with T₃ is not an effective means of stimulating growth in red drum fed optimum diets. Whereas thyroid hormones may function to regulate intermediary metabolism in red drum, elevated endogenous thyroid hormone levels appear adequate to supply tissue needs during juvenile growth in culture.     

美国红鱼养殖种群的同工酶酶谱及其生化遗传初步分析

利用水平淀粉凝胶电泳技术，对50尾美国红鱼肌肉及肝脏组织进行了14种同工酶的初步分析。结果表明，美国红鱼多态位点比例P＝0.292，实际平均杂合度Hp=0.057，预期杂合度He=0.108，每个位点的有效等位基因数Ae=1.108，Hardy-Weinberg遗传偏离指数D＝－0.472。     

浙江自然海区逃逸美国红鱼食性研究

通过对浙江自然海区逃逸美国红鱼胃含物的分析,从食料生物组成、食性类型、摄食强度及营养级4个方面对浙江自然海区逃逸美国红鱼的食性进行了探讨。结果表明,美国红鱼属游泳动物食性。主要摄食鱼类、长尾类、短尾类和头足类,兼食口足类和多毛类。其食料生物组成和食性类型与原栖息地有较大差异。逃逸美国红鱼的摄食率为80.28%,最高饱满度系数达60.93‰,平均10.26‰。根据计算,其饵料的生境宽度为4.09,营养级为3.43级。浙江自然海区美国红鱼是一种凶猛的中级肉食性鱼类。     

Evaluation of fisheries by-catch and by-product meals in diets for red drum Sciaenops ocellatus L.

This study evaluated various by‐catch and by‐product meals of marine origin with red drum (Sciaenops ocellatus L.). Four different kinds of by‐catch or by‐product meals [shrimp by‐catch meal from shrimp trawling, Pacific white shrimp (Litopenaeus vannamei (Boone)) processing waste meal, red salmon (Oncorhynchus nerka (Walbaum)) head meal, and Pacific whiting (Merluccius productus (Ayres)) meal] were substituted for Special Select? menhaden fish meal at 33% or 67% of crude protein in diets formulated to contain 40% crude protein, 12% lipid, and 14.6 kJ digestible energy g^?1. Each of these diets and three additional diets consisting of shrimp processing waste meal formulated on a digestible‐protein basis and two Pacific whiting diets containing reduced levels of ash were also evaluated in two 6‐week feeding trials with juvenile red drum (initial weight of 4–5 and 1–2 g fish^?1 in trials 1 and 2). Red drum fed by‐catch meal at either level of substitution performed as well as fish fed the control diet; whereas, fish fed shrimp processing waste meal diets had significantly (P≤0.05) reduced weight gain and feed efficiency ratio values compared with the controls, even when fed on a digestible‐protein basis. The diets containing Pacific whiting at either levels of substitution and regardless of ash level supported similar performance of red drum as those fed the control diet. Fish fed the red salmon head meal diet fared poorly, probably owing to an excessive amount of lipid in the diet that became rancid. Overall, by‐catch meal associated with shrimp trawling and Pacific whiting appear to be suitable protein feedstuffs for red drum.     

Essential fatty acid requirement of juvenile red drum (Sciaenops ocellatus)

Feeding experiments and laboratory analyses were conducted to establish the essential fatty acid (EFA) requirement of red drum (Sciaenops ocellatus). Juvenile red drum were maintained in aquaria containing brackish water (5 ± 2‰ total dissolved solids) for two 6-week experiments. Semipurified diets contained a total of 70% lipid consisting of different combinations of tristearin [predominantly 18:0] and the following fatty acid ethyl esters: oleate, linoleate, linolenate, and a mixture of highly unsaturated fatty acids (HUFA) containing approximately 60% eicosapentaenoate plus docosahexaenoate. EFA-deficient diets (containing only tristearin or oleate) rapidly reduced fish growth and feed efficiency, and increased mortality. Fin erosion and a “shock syndrome” also occurred in association with EFA deficiency. Of the diets containing fatty acid ethyl esters, those with 0.5–1% (n-3) HUFA (0.3–0.6% eicosapentaenoate plus docosahexaenoate) promoted the best growth, survival, and feed efficiency; however, the control diet containing 7% menhaden fish oil provided the best performance. Excess (n-3) HUFA suppressed fish weight gain; suppression became evident at 1.5% (n-3) HUFA, and was pronounced at 2.5%. Fatty acid compositions of whole-body, muscle and liver tissues from red drum fed the various diets generally reflected dietary fatty acids, but modifications of these patterns also were evident. Levels of saturated fatty acids appeared to be regulated independent of diet. In fish fed EFA-deficient diets (containing only tristearin or oleate), monoenes increased and (n-3) HUFA were preferentially conserved in polar lipid fractions. Eicosatrienoic acid [20:3(n-9)] was not elevated in EFA-deficient red drum, apparently due to their limited ability to transform fatty acids. Red drum exhibited some limited ability to elongate and desaturate linoleic acid [18:2(n-6)] and linolenic acid [18:3(n-3)]; however, metabolism of 18:3(n-3) did not generally result in increased tissue levels of (n-3) HUFA. Based on these responses, the red drum required approximately 0.5% (n-3) HUFA in the diet (approximately 7% of dietary lipid) for proper growth and health.     

Digestive enzyme activity during larval development of the Senegal sole (Solea senegalensis)

The activities of the main digestive enzymes (proteases, amylase, lipase) as well as those of acid and alkaline phosphatases were assessed during the larval development of the Senegal sole, Solea senegalensis. Important variations in specific activities of all the enzymes were observed during the period of study and were mostly related to the beginning or the end of metamorphosis. Both acid and alkaline protease activities were identified at early stages of development. Acid proteases accounted for only 10% of total protease activity in a 9 days-old larva, but exceeded 75% in a 33 days-old individual. Maximum lipase activity was related to the development of exocrine pancreas (days 6 to 10) and to metamorphosis. It is suggested this can be related to the metabolism of lipid reserves taking place during this morphological change. Activity of alkaline phosphatase decreased from day 5 to day 20 but therafter revealed a sharp increase, possibly linked to the development of enterocytes. The pattern of development of the main digestive enzymes found in S. senegalensis is similar to that described in other flatfish species.     

Effects of dietary thyroid hormones on the red drum (Sciaenops ocellatus)

Four separate 8-week feeding trials were conducted to assess the effects of supplementing semipurified diets with either triiodothyronine (T₃) or thyroxine (T₄) at 0, 2, 10, and 50 mg/kg on growth and body composition of juvenile red drum (Sciaenops ocellatus) held in artificial brackish water (6‰) and artificial seawater (32‰). At both levels of salinity, increasing doses of T₃ resulted in fish with reduced weight gain, feed efficiency, condition factor (weight × 100/length³), and muscle ratio (muscle weight × 100/body weight), as well as a lighter body color. Significant (p < 0.05) effects of T₃ on the proximate composition of whole body, liver, and muscle were variable, generally reflecting decreased lipid and protein storage in liver and muscle, respectively. The two highest doses of T₃ given to seawater adapted fish increased survival. Dietary T₄ supplementation had no distinctive effects on appearance, growth or proximate body composition. These results indicate that whereas T₃ may function to regulate protein and lipid metabolism in red drum, dietary supplementation with T₃ leads to a hyperthyroidism-induced catabolic state. The elevated endogenous thyroid hormone levels found in fish fed optimal diets may thus adequately supply tissue needs during juvenile growth.     

Evaluation of dietary creatine and guanidinoacetic acid supplementation in juvenile red drum Sciaenops ocellatus

Two separate comparative feeding trials were conducted to evaluate if supplemental dietary guanidinoacetic acid (GDA), either singly or in combination with creatine, could enhance growth performance of red drum. The basal diet for both trials was formulated with practical ingredients but was not supplemented with creatine or GDA. For the experimental diets, creatine (0 or 20 g/kg) and GDA (0, 5 or 10 g/kg) were added to the basal diet in a 2 × 3 factorial arrangement for trial 1. Another 2 × 3 experimental design was adopted in trial 2 to further evaluate creatine (0 or 20 g/kg) and GDA (0, 10 or 20 g/kg) supplementation. Each diet was fed to juvenile red drum in either quadruplicate (trial 1) or triplicate (trial 2) aquaria twice daily for a total of 8 weeks. After each feeding trial, fish were sampled for body condition indices and whole‐body composition analysis, as well as determination of plasma, liver and muscle creatine concentrations. In trial 1, dietary creatine supplementation alone significantly (p < .05) improved weight gain and feed efficiency of red drum. An interaction between creatine and GDA was seen in whole‐body protein and lipid in the two trials. In trial 1, the highest levels of whole‐body protein and lipid were observed in fish fed 10 g GDA/kg, and in trial 2, supplementation of the diet with 10 g GDA/kg increased muscle protein and muscle lipid although there was a trending decline in fish fed the 20 g/kg GDA‐supplemented diet. In both trials of the present study, dietary GDA significantly (p < .05) enhanced liver creatine content. Dietary creatine also significantly (p < .05) enhanced plasma and muscle creatine content of red drum. Based on the results of this study, creatine was effective in enhancing weight gain and feed efficiency of red drum as seen in previous studies; however, GDA was not effective in influencing growth performance but limited synthesis of creatine from dietary GDA was apparent.     

Gill morphometry of the red drum,Sciaenops ocellatus

The structure and morphometry of the gills of the marine teleost, red drum, have been studied. The present analysis of gas exchange area of fish gills is one of the most intensive and the results are compared to less intense averaging methods. Based on the gill area estimates, red drum falls into the category of a fish of intermediate activity. Its gill clearly has an exchange area less than that of the tunas, but is slightly greater than that of trout or bass. The three components that contribute to total exchange area (filament length, lamellar density, and area of individual lamellae) are not all greater in species with a greater total exchange area. The best correlate is total filament length.     

Sex differences in circulating steroid hormone levels in the red drum, Sciaenops ocellatus L.

Steroid profiles of cultured and captive red drum (Sciaenops ocellatus L.) were investigated to evaluate the potential use of circulating sex steroid levels as a tool for gender identification in this species. Cultured 18‐month‐old fish were maintained on a 120‐day shortened photothermal cycle to induce precocious maturation. Additionally, wild‐caught fish were maintained in captivity under simulated natural photothermal conditions from late spring to early fall. Circulating 11‐ketotestosterone (11‐KT) levels were significantly higher in males compared with females during the early stages of gonadal growth in both cultured and captive fish. Plasma testosterone (T) levels showed a similar trend; however, the differences were significant only when males were already producing sperm. 17β‐estradiol (E₂) concentrations were low in males and females before gonadal recrudescence but increased significantly with the progression of vitellogenesis in females. These results show that a test using a minimum concentration of circulating 11‐KT could be developed to differentiate between sexes in the early stages of gonadal maturation in red drum. Moreover, plasma E₂ concentrations could be used to identify vitellogenic females. The two steroids considered together could help avoid possible error in gender identification due to unusually high levels of certain steroids encountered in some individuals.