Optimized Extraction,Preliminary Characterization and Evaluation of the in Vitro Anticancer Activity of Phlorotannin-Rich Fraction from the Brown Seaweed,Cystoseira sedoides

Brown seaweeds produce useful bioactive substances with high cosmetic and pharmacological values due to the presence of antioxidant derivatives, mainly phlorotannins (PHT), which are of particular interest. A central composite rotatable design (CCRD) with three variables (extraction time, dry material-to-solvent ratio and ethanol concentration) and two responses was performed to optimize the microwave-assisted extraction (MAE) of phlorotannins from Cystoseira sedoides using response surface methodology (RSM). Results showed that the independent variables significantly affected both phlorotannin content and the scavenging capacity. The optimum operating conditions were extraction time, 101.74 sec; dry material-to-solvent ratio, 1:10 g/mL; and ethanol extraction, 50%. Under these conditions, the predicted values of PHT content and radical scavenging activity-IC50 were close to the observed values and were 383.887 µg PGE/g Dm and 18.353 µg/mL, respectively. Characterization of the phlorotannin-rich fraction was conducted by high-performance liquid chromatography and Fourier transform infrared spectroscopy. The evaluation of the anticancer activity against MCF-7 breast cancer cell line showed a potent activity to trigger apoptotic death in more than a half of the MCF-7 cells, with an estimated IC50 value of 78 μg/mL. In addition, this fraction induced a notable growth regression effect on 3D spheroids model in a concentration-dependent manner, with a growth rate of about 1.17, at 200 µg/mL.

Abbreviations: CCRD: Central composite rotatable design; Dm: Dry material; DMBA: 2,4-dimethoxy benzaldehyde; DMSO: Dimethyl sulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTA: Ethylene diamine tetraacetic acid; FBS: Fetal bovine serum; FT-IR: Fourier transform infrared; HPLC: High-performance liquid chromatography; IC₅₀: Half-maximal inhibitory concentration; MAE: Microwave-assisted extraction; Min: minutes; PBS: Phosphate buffered saline; PGE: Phloroglucinol equivalent; PHT: Phlorotannins; PHT-SED: phlorotannins derived from Cystoseira sedoides; PI: Propidium iodide; RSA: Radical scavenging activity; RSM: Response surface methodology; Sec: seconds     

Optimal Immobilization of Acidic Proteases from Monterey Sardine (Sardinops sagax caeurelea) on Partially Deacetylated Chitin from Shrimp Head Waste

ABSTRACT

Response surface methodology was employed to optimize the immobilization yield of acidic proteases from Monterey sardine (Sardinops sagax caeurelea) using partially deacetylated chitin as immobilization support. A rotatable central composite design was applied to evaluate the effects of immobilization conditions such as enzyme loading (X₁), immobilization pH (X₂), and tripolyphosphate concentration (X₃) on the immobilization yield. The analysis of variance revealed that the established model was significant (p < 0.05), and the adjustment of the quadratic model with the experimental data was satisfactory. Under optimal conditions (X₁ = 0.05 mg/mL, X₂ = 3.16, and X₃ = 0.75%), an immobilization yield of 79.1% was achieved; a value that was in accordance with the predicted one.     

星康吉鳗形态性状与体重的通径分析及生长曲线拟合

为探明星康吉鳗（Conger myriaster）形态性状和体重的关系,测定了体重（Y）及全长（X₁）、肛长（X₂）、尾长（X₃）、头长（X₄）、吻长（X₅）、眼径（X₆）、眼后头长（X₇）、背鳍前长（X₈）、躯干长（X₉）、头宽（X₁₀）、眼间距（X₁₁）等11个形态性状,通过相关分析、通径分析、回归分析等方法研究了形态性状对体重的影响;通过曲线拟合获得4个形态性状与体重的最佳拟合模型。结果表明,星康吉鳗各形态性状之间以及形态性状与体重之间均呈极显著正相关关系（P< 0.01）。相关分析发现肛长（X₂）与体重的相关系数最大（0.970）;通径分析发现肛长（X₂）对体重的直接作用最大（0.450）,头宽（X₁₀）通过肛长（X₂）对体重的间接作用最大（0.431）;决定系数分析发现,肛长（X₂）对体重的直接决定系数最大（0.203）,肛长（X₂）和头宽（X₁₀）的共同决定系数最大（0.172）。形态性状（X）与体重（Y）的多元回归方程为Y=-113.859+6.481X₂+49.213X₆+11.514X₇+20.145X₁₀（R²=0.954）。星康吉鳗肛长（X₂）、眼后头长（X₇）、头宽（X₁₀）与体重的最佳拟合模型为指数函数,模型方程分别为Y=1.466e^0.287X、Y=1.435e^1.328X、Y=1.970e^1.974X,眼径（X₆）与体重的最佳拟合模型为线性,方程为Y=-77.460+222.093X。结论认为,在星康吉鳗选育时,应以肛长（X₂）和头宽（X₁₀）为主要选择性状,以眼径（X₆）和眼后头长（X₇）为辅助选择性状。     

四角蛤蜊形态性状对重量性状的影响效果分析

以随机选取的123只野生四角蛤蜊为材料,对其形态性状(壳长X₁、壳高X₂、壳宽X₃)和重量性状(活体质量Y、软体质量Z)进行了测量,然后运用相关分析、通径分析和回归分析等分析方法研究形态性状对重量性状的影响。结果表明,5个数量性状间的相关系数均达到极显著水平(P<0.01)。形态性状对活体质量和软体质量的直接影响依次为壳宽>壳长>壳高。壳宽是影响活体质量和软体质量的主要因素,其决定系数分别为0.302 5和0.224 7。多元回归分析建立了估计活体质量、软体质量的最优回归方程:Y=-19.537+0.244X₁+0.266X₂+0.732X₃、Z=-2.456+0.035X₁+0.035X₂+0.089X₃。     

Marking live feeds with inert metal oxides for fish larvae feeding and nutrition studies

Yttrium oxide (Y₂O₃), ytterbium oxide (Yb₂O₃), lanthanum oxide (La₂O₃) and dysprosium oxide (Dy₂O₃) were evaluated as potential live feed markers for feeding and nutrition studies with fish larvae, by determining the uptake and depletion of markers over time in two trials, and quantifying ingestion of Y₂O₃‐marked rotifers (Branchionus plicatilis) by Atlantic cod (Gadus morhua) in a third trial. In the first two trials, Artemia nauplii and rotifers quickly took up markers within 10 min to concentrations useful for nutrition studies (>2% dry weight). There was no significant difference (P>0.05) among temperatures in depletion of markers (10, 15, 20 °C) with Artemia or rotifers. Depletion from rotifers was not significantly different (P>0.05) between 5 and 20 min nor between 5 and 30 min for Artemia when marked at a concentration of 50 mg of marker per litre of seawater. In the second trial, rotifers and Artemia were marked with a higher concentration (250 mg L^?1) and allowed to deplete for a longer time (90 min). In the third trial, visual estimates of Artemia consumed by Atlantic cod larvae were similar to consumption estimates determined by analysis of Y₂O₃‐marked Artemia using inductively coupled plasma optical emission spectroscopy (r²=0.77).     

不同时期牙鲆形态性状对体重影响的通径分析及曲线拟合研究

以8月龄与14月龄的养殖牙鲆(Paralichthys olivaceus)为研究对象,分别测定体重(Y)和22个形态性状,对各性状进行相关分析,并建立多元回归方程,对各形态性状对体重的影响效果进行通径分析。分别以进入回归方程的各形态性状作为自变量,体重为因变量进行曲线模型拟合,筛选最优拟合模型。结果显示:(1)在不同生长阶段,相同形态性状与体重的相关性存在差异。(2)8月龄阶段,X_(18)(腹鳍基部到背鳍终点的直线距离)、X_4(体宽)和X_(11)(背鳍起点到臀鳍起点的直线距离)对体重的通径系数达到极显著水平(P0.01),14月龄阶段,X_(18),X_(14)(背鳍终点到臀鳍终点的直线距离)和X_9(尾柄长)对体重的通径系数达到显著水平(P0.05),由此可知,不同生长阶段,影响体重的形态性状不尽相同。8月龄和14月龄牙鲆形态性状对体重的多元回归方程分别为:Y=-119.541+7.191X_(18)+10.135X_4+7.197X_(11);Y=-484.931+31.959X_(18)+81.928X_(14)-17.899X_9。(3)8月龄阶段,将回归模型中的3个自变量分别与体重进行模型拟合,最优拟合模型均为线性模型,分别为:Y=-117.866+15.724X_(18);Y=-94.579+24.763X_4;Y=-100.602+33.184X_(11);14月龄阶段,将回归模型中的3个形态性状分别与体重进行模型拟合,最优拟合模型均为幂函数模型,分别为:Y=0.036X_(18)~(3.063);Y=0.095X_(14)~(2.587);Y=62.249X_9~(1.584)。本次研究表明,在牙鲆的不同生长时期,影响体重的主要形态性状不同,两阶段适用的最优拟合模型也不同,这为牙鲆不同时期的选择育种工作提供了理论依据。     

An evaluation of phytase for Channel catfish (Ictalurus punctatus) fed all plant‐protein diet: Growth performance,nutrient utilization and P equivalency value

This study evaluated the effect and phosphorus equivalency value of dietary microbial phytase based on growth performance and nutrient utilization of Channel catfish (Ictalurus punctatus). An all plant‐protein basal diet containing 7.9 g/kg total Phosphorus (P) (Phytate P of 4.5 g/kg) was supplemented with graded levels 0, 300, 500, 1,000, 1,500 and 2,000 U phytase/kg of diet or with 3, 5, 8, 12 g/kg of Ca(H₂PO₄)₂, namely with 0.7, 1.2, 2.0, 3.09 g/kg P as Ca(H₂PO₄)₂. Triplicate groups of Channel catfish (Ictalurus punctatus) juveniles (initial weight, 1.70 ± 0.04 g) were fed the experiment diets for 90 days. The results indicated that dietary phytase supplementation significantly improved growth performance, apparent digestibility coefficient of P and P retention, and decreased feed conversion ratio (FCR), faecal P content and P excretion per gain. Line relation was found between the effects of P and phytase supplementation levels, and the linear response equations of the effects of P and phytase levels were generated and used to calculate the P equivalency values. The average function of P equivalency values (Y, g/kg) of microbial phytase (X, U/kg of diet) was developed: Y = 0.1695 + 0.0021X; R² = 0.996. The results showed that about 0.38 g of P/kg diet was released by per 100 U of phytase over the range of 300–2,000 U of phytase/kg diet for Channel catfish (Ictalurus punctatus) fed all plant‐protein diet.     

Pharmacokinetics and tissue residues of marbofloxacin in crucian carp (Carassius auratus) after oral administration

Pharmacokinetics and residue elimination of marbofloxacin (MBF) were studied in crucian carp (Carassius auratus, 250±30 g) kept at two water temperatures of 15 and 25 °C. Marbofloxacin concentrations in plasma and tissues were analysed by means of high‐performance liquid chromatography using an ultraviolet detector. The limits of detection were 0.02 μg mL^?1, 0.02 μg g^?1, 0.025 μg g^?1, 0.02 μg g^?1 and 0.025 μg g^?1 in plasma and muscle, skin, liver and kidney respectively. Fish were administered orally at a single dosage of 10 mg kg^?1 body weight in the PK group. The data were fitted to two‐compartment open models at both temperatures. At 15 °C, the absorption half‐life () and distribution half‐life (t_1/2α) of the drug were 0.36 and 4.48 h respectively. The corresponding values at 25 °C were 0.23 and 0.87 h respectively. The elimination half‐life (t_1/2β) was 50.75 h at 15 °C and 25.05 h at 25 °C. The maximum MBF concentration (C_max) differed little between 15 (6.43 μg mL^?1) and 25 °C (8.36 μg mL^?1). The time to peak concentration was 1.74 h at 15 °C and 0.78 h at 25 °C. The apparent volume of distribution (V_d/F) of MBF was estimated to be 1.36 and 0.87 L kg^?1 at 15 and 25 °C respectively. The area under the concentration–time curve (AUC) was 301.80 μg mL^?1 h at 15 °C and 182.80 μg mL^?1 h at 25 °C. The total clearance of MBF was computed as 0.03 and 0.05 L h^?1 kg^?1 at 15 and 25 °C respectively. After repeated oral administration at a dosage of 10 mg kg^?1 body weight per day for 3 days, the results showed that the elimination half‐lives () of MBF from all tissues at 15 °C were longer than that at 25 °C. Therefore, water temperature is an important factor to be considered when deciding a reasonable withdrawal time.     

臭氧对条石鲷幼鱼鳃组织结构和抗氧化系统的影响

臭氧(O₃)在工厂化养殖水处理中有较高的应用价值,为了解臭氧对条石鲷幼鱼的影响,采用自制的臭氧毒性试验装置(授权利号:ZL 200920122441.3),对条石鲷幼鱼进行了不同质量浓度(0、0.08、0.13、0.17、0.22、0.28和0.32 mg/L)和不同时间(0.5、1、6、12、18和24 h)的急性胁迫试验,并分析了臭氧对条石鲷幼鱼的行为响应、半致死浓度、幼鱼鳃的组织形态学、酶活性等的影响。结果表明,随着臭氧胁迫浓度的升高,条石鲷幼鱼在水层中的分布位置逐渐上移,呼吸频率逐渐加快,受惊反应逐渐迟缓;条石鲷幼鱼的开始死亡时间与臭氧浓度负相关,12 h的半致死方程为Y=16.815 8+20.340 5X(r=0.998 7),半致死浓度为0.262 5 mg/L。24 h的半致死方程为Y=17.614 1+17.898 3X(r=0.950 0),半致死浓度为0.197 4 mg/L;条石鲷幼鱼鳃SOD和GPX活性与臭氧胁迫时间和胁迫浓度负相关,并都在胁迫24 h、质量浓度0.32 mg/L时降至谷值;MDA含量与臭氧胁迫时间和胁迫浓度正相关,并在胁迫24 h、质量浓度0.32 mg/L时达到峰值;在臭氧胁迫下,条石鲷幼鱼的鳃小片细胞排列不规则并萎缩,鳃小片上皮细胞脱落或粘滞于鳃小片间,鳃小片末端膨大,鳃丝血管固缩无腔或变空腔,管壁粗糙、干缩,血细胞固缩、坏死,零散分布或紧贴血管壁。     

Influence of fish size and water temperature on the metabolic demand for oxygen by barramundi, Lates calcarifer (Bloch), in freshwater

Oxygen demand by all animals is driven primarily by their needs for sustaining metabolism. Typically, larger animals require more oxygen and cellular fuel to carry out respiration than smaller animals. This relationship in most cases is not linear and is usually described by a coefficient and exponent (e.g. ax^b): the exponent b showing the relationship between live‐weight and energy/oxygen demand and is often termed the metabolic body weight (MBW) exponent, while the coefficient (a) tends to be temperature specific and describes the relationship between MBW and maintenance metabolic energy and oxygen demand at that specific temperature. Across all temperatures (range 26.0–32.0°C), the relationship between barramundi (Lates calcarifer) live‐weight (x; g) and relative oxygen consumption as standard metabolic rate (y; mg O₂ kg^?1 h^?1) at 29.4±1.5°C (mean±SD) was described by the exponential curve: y=710.19 x^?0.3268, R²=0.6875 (n=222 assessments). Examination of the same data but on a gross oxygen consumption (mg O₂ h^?1) basis showed a relationship between live‐weight (x; g) and gross oxygen consumption (y; mg O₂ h^?1) that was described by the exponential curve: y=0.710 x^0.6732, R²=0.9033. Evaluation of the combined relationship between fish live‐weight (y; g) and water temperature (x; °C) on gross oxygen consumption rate (z; mg O₂ h^?1) was described by the equation: z=(?20.7818+1.4017x?0.0227x²) ×y^0.673.     

Effect of dietary protein level on growth and food utilization in juvenile Murray cod Maccullochella peelii peelii (Mitchell)

Murray cod Maccullochella peelii peelii (Mitchell) is a freshwater Percichthyid fish considered to have high culture potential. Growth and feed utilization were examined in a 56‐day experiment, in which triplicate groups of juvenile Murray cod (initial weight 21.5 ± 0.03 g) were fed isocalorific diets (gross energy content of about 21 kJ g^?1) containing 40%, 45%, 50%, 55% or 60% protein (designated P40, P45, etc.). Final mean weight, percentage increase in weight and specific growth rate (SGR; % day?1) were highest in fish fed the P50 diet. Food conversion ratio (FCR; 1.05 ± 0.04) and protein efficiency ratio (PER; 1.98 ± 0.11) were also best in fish on the P50 diet, but the differences in these parameters from the corresponding values on diets P55 and P60 were not always significant. FCR (Y) was related to dietary protein content (X), the relationship being a second‐order polynomial, in which Y = 0.004X²?0.431X + 12.305 (r= 0.95; P < 0.01). The proportions of carcass moisture, protein, lipid and ash did not differ among the different dietary treatments. The protein conversion efficiency (y) was negatively correlated to percentage dietary protein (X) content, the relationship being: y = 62.76–0.62X (r= 0.99; P < 0.01).     

Effects of temperature on food consumption, growth and oxygen consumption of freshwater prawn Macrobrachium rosenbergii (de Man 1879) postlarvae

Effects of temperature on food consumption, growth and oxygen consumption were estimated for the freshwater prawn Macrobrachium rosenbergii postlarvae at 23 °C, 28 °C and 33 °C in the laboratory. The results showed that the animal's initial body weight had a close linear relationship with food consumption and growth. Food consumption increased directly with temperature. Consumption rates (C; mg day^?1 ind^?1 ) of the 28 °C and 33 °C groups were much higher than that of the 23 °C group (P < 0.001), and the 33 °C group's consumption rate was higher than that of the 28 °C group (P < 0.05). The relationship of food consumption with temperature and initial body weight (W; mg) could be described as: C = 0.0679W + 0.185t? 3.17. Growth increased significantly with increased temperature. The relationship among specific growth rate, temperature and initial body weight was as follows: SGR = ?0.110W + 0.213t + 0.176. However, temperature showed no effect on growth efficiency. Oxygen consumption increased significantly with temperature (P < 0.01). The weight‐specific oxygen consumption rates (mg O₂ g^?1 h^?1) at 23 °C, 28 °C and 33 °C were 0.83, 1.16 and 1.49 mg O₂ g^?1 h^?1 for 61.92 mg M. rosenbergii.     

ACUTE TOXICITY OF NITRITE TO SEA BASS (Dicentrarchus labrax) AND EUROPEAN EEL (Anguilla anguilla)

Static acute toxicity tests of 4 and 7 days duration were performed on sea bass (Dicentrarchus labrax) and European eel (Anguilla anguilla). The 96-hour LC₅₀ for sea bass (x?; = 5 cm) was 274, 220, and 154 mg/liter N-NO₂ at water temperatures of 17, 23 and 27°C, respectively, in sea water. The 96-hour LC₅₀ for eel elvers was 84, 213, 264, 503, 812 and 974 at salinities of 0, 6, 12, 24, 30 and 36 g/liter.     

A Length–Weight Relationship for Pond‐raised Hybrid Catfish Fingerlings

A length–weight relationship was developed for pond‐raised hybrid catfish fingerlings, Ictalurus punctatus × Ictalurus furcatus. A total of 4663 fish were sampled ranging from 20 to 240 mm total length. Predicted weight (g) from length (mm) is based on the equation: Y = 0.000017311X^2.868474309 (r² = 0.996) where Y is the estimated individual fish weight and X is measured total length. The predicted values generated by this model differ from, but follow the same general trends seen with channel catfish, I. punctatus, fingerlings and can be used as a baseline for the development of a length–weight table for hybrid catfish fingerling production.     

Pharmacokinetic disposition of enrofloxacin in brown trout (Salmo trutta fario) after oral and intravenous administrations

In this study, the pharmacokinetic profile of enrofloxacin (EF) and its major metabolite, ciprofloxacin (CF), were investigated in brown trout (Salmo trutta fario) (n = 150) after intravenous (i.v.) and oral (p.o.) administrations of a single dose of 10 mg kg^− 1 body weight (b.w.) at 10 °C. The plasma concentrations of the drugs were determined by high-performance liquid chromatography (HPLC-UV) from 0.08 to 120 h. Pharmacokinetic parameters were described by the two-compartment open model for intravenous and oral administrations, respectively. After intravenous administration, the elimination half-life (t_1/2β), apparent volume of distribution at steady-state (V_ss) and total body clearance (Cl_tot) of enrofloxacin were 19.14 ± 1.51 h, 3.40 ± 0.18 L kg^− 1 and 0.14 ± 0.01 L kg h^− 1, respectively. After oral administration, the maximum plasma concentration (C_max), time of maximum concentration (t_max) and bioavailability (F%) were 2.30 ± 0.08 µg mL^− 1, 8 h and 78 ± 4%, respectively. Ciprofloxacin was not detected in the present study. The elimination half-life for enrofloxacin following oral administration was longer than values calculated for other animals. After oral administration, the mean plasma concentration was well above the minimum inhibitory concentrations (MICs)—that is, > 0.5 µg mL^− 1 at 36 h—for most gram-negative fish pathogens. It is possible and practical to obtain therapeutic blood concentrations of enrofloxacin in brown trout (S. trutta fario) using oral administration of 10 mg kg^− 1 body weight; therefore, it may be effective in the therapy for brown trout diseases.     

Reproductive biology of Oreochromis mossambicus populations of man-made lakes in Sri Lanka: a comparative study

Abstract. The sex ratio, mean size at maturity (L_m), gonosomatic index (GSI), frequency of distribution of egg diameter of the mature ovaries and fecundity of Oreochromis mossambicus (Peters) populations of 12 man-made lakes were studied using commercial landings from the gill-net fishery. The overall sex ratio was 1·0:0·88 of females to males and that of the individual populations varied from 1·0:0·54 to 1·0:1·34. The L_m ranged from 16·0 to 21·0 cm (TL) and was found to be correlated to the size of the water-body. The GSI of the females in stage V, the stage prior to spawning, did not differ from each other, except in one reservoir. The variations in the mean egg size (μ) of different populations were negatively correlated to the fecundity of a Ash of standard weight (FW₂₀₀) of the different populations and the relationship was, y =3487·4– 1·14FW₂₀₀(df = 10; r= -0·64; P < 0·05) The overall fecundity ranged from 318 to 3169 in fish ranging in body length (TL) and body weight from 16·5 to 32·0 cm and 80 to 651 g respectively. Fecundity was linearly related to body weight and curvilinearly to body length. The exponent b of the fecundity (F) length (1) regressions (F = aL^b) of the different populations varied between 1·20 and 3·83. The overall relationship of fecundity to body weight (W) and body length (1) for O. mossambicus reservoir populations in Sri Lanka were: F = 3·23W + 357·8 (df = 203; r= 0·73; P < 0·001) F = l·52L^2·11(df = 203; r= 0·68; P < 0·001) The variations in the theoretical fecundity calculated for a standard size individual of 200 g in weight (FW₂₀₀) of the reservoir populations were correlated to the mean fishing pressure (X) exerted on each population according to the equations. (a) FW₂₀₀= 10–24X+ 896·8 (df 9; r= 0·58; P < 0·05) (b) FW₂₀₀= 1200 – 422·76e^?0·102X (df 9; P < 0·05)     

Optimization of Antioxidant Activity and Phenolic Compound Extraction Conditions from Red Seaweed (Laurencia obtuse)

ABSTRACT

Response surface methodology was used to investigate the effect of ultrasound-assisted extraction conditions including solvent:seaweed ratio (10:1–30:1), extraction temperature (30–50°C), and extraction time (30–60 min) on the total phenolic compounds and antioxidant activity of red seaweed (Laurencia obtuse) extracts. The optimum extraction parameters for maximum phenolic content were as follows: solvent:seaweed ratio, 30:1; extraction temperature, 50°C; and extraction time, 42.8 min. The experiment results showed that the solvent:seaweed ratio was the most significant parameter for the extraction. The optimum extraction conditions for maximum antioxidant activity were as follows: solvent:seaweed ratio, 24.3:1; extraction temperature, 45.3°C; and extraction time, 58 min. Under the above-mentioned conditions, the experimental total phenolic content and antioxidant activity value were 26.23 ± 0.75 mg GAE/g seaweed and 120.89 ± 0.81 TEAC, respectively, of seaweed tested, which are well compatible with the predicted contents.     

Standardization of photometric measurement of sperm concentration from diploid and tetraploid Pacific oysters, Crassostrea gigas (Thunberg)

To provide necessary standardization of procedures for cryopreservation of sperm, a spectrophotomeric method was developed to determine the sperm concentration of diploid and tetraploid Pacific oysters, Crassostrea gigas. Wavelengths of 380, 550, 581 and 780 nm were compared, and 550 and 581 nm were found to be the most sensitive and reliable. A linear relationship between sperm concentration and photometric absorbance was observed for sperm concentrations between 2 × 10⁷ and 2 × 10⁹ cells mL^?1. The regression equation for the standard curve at 550 nm for sperm of diploid oysters was Y=?8.528+1.165 log X. The equation for sperm of tetraploid oysters was Y=?8.844+1.236 log X. The equation at 581 nm for sperm of diploid oysters was Y=?8.07+1.104 log X. The equation at 581 nm for sperm of tetraploid oysters was Y=?8.331+1.167 log X. Comparisons derived from the standard curves at 581 nm between observed values and the predicted values indicated good agreement for sperm from diploid (coefficient of determination, r²=0.983) and tetraploid (r²=0.980) oysters.     

Interactions between temperature and size on the growth,size heterogeneity,mortality and cannibalism in cultured larvae and juveniles of the Asian catfish,Pangasianodon hypophthalmus (Sauvage)

Temperature effects vary throughout the ontogeny, but are proportionally more variable during the early life stages than in older fish. The larvae of a few species have been studied but contrasting trajectories were observed, thereby making it difficult to predict how temperature impacts on the growth, survival and size heterogeneity in a particular species. This study examined these interactions in young [0.9 mg–1 g wet mass (WM)] Pangasianodon hypophthalmus, one of the most extensively cultured tropical fish. Fish were raised at five temperatures from 23 to 33 °C in a recirculated water system, fed in excess with a high‐energy feed (Artemia then a formulated feed, >50% protein), and examined at 1‐day (first feeding days) or 4‐day intervals (older fish). The temperature that produced the fastest growth (T°_opt) was 31 °C at the start of exogenous feeding (0.9 mg); it increased to 32.7 °C at 8 mg then decreased by 0.7 °C for each 10‐fold increase of WM. Size heterogeneity was lower and survival was higher (70–85% from 0.9 mg to 1 g) when the temperature was close to T°_opt. Comparisons with other species suggest that the slope of the relationship between T°_opt and fish size has a strong latitudinal component, and may also reflect the fish sensitivity to oxygen.