Pharmacokinetics of Eugenol in Japanese Flounder,Paralichthys olivaceus

Eugenol, which has been used as an anesthetic for fish, was administered to Japanese flounder, Paralichthys olivaceus (127 ± 50.8 g), by bath treatment at concentrations of 0.025, 0.125, and 0.25 mL/L seawater for 10 min and intramuscular injection at 40 μL/fish to investigate the pharmacokinetic characteristics of eugenol in the plasma of Japanese flounder. In the bath treatment test, plasma eugenol concentration increased with increasing eugenol concentration up to 0.125 mL/L and reached steady state within 5 min. After a 10‐min bath treatment in 0.25 mL/L eugenol, plasma eugenol concentration was about 58.4 µg/mL. After transfer into running seawater, plasma eugenol concentration decreased biphasically with half‐lives of 0.0296 h (α‐phase) and 0.289 h (β‐phase). The AUC_0000→0800 was about 16.5 µg h/mL. In administration by intramuscular injection, plasma eugenol concentration increased rapidly after administration and decreased biphasically with half‐lives of 0.0329 h (α‐phase) and 8.08 h (β‐phase). The AUC_0000→0800 was about 52.5 µg h/mL. In both methods of administration, Japanese flounder with average weight of 127 g were effectively anesthetized when plasma eugenol concentrations were between 2.19 and 4.88 µg/mL.     

Effects of Fish Size and Water Temperature on the Acute Toxicity of Copper for Japanese Flounder,Paralichthys olivaceus,and Red Sea Bream,Pagrus major

The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.     

Survival and Growth Improvement of Palm Ruff,Seriolella violacea,Larvae Fed Artemia Nauplii Enriched with an Experimental Emulsion

The palm ruff, Seriolella violacea (Cojinoba), is a potential new species for Chilean aquaculture. To approach Cojinoba larviculture, an experimental Artemia enrichment emulsion, containing docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) = 2.5, supplemented with vitamin E, astaxanthin, and β‐glucan, was evaluated in both Artemia and Cojinoba larvae, 30–50 d.a.h. This study tested an experimental enrichment emulsion versus a commercial emulsion, with an integral approach of multicompound emulsions. After 23 h enrichment, experimental emulsion (EE)‐enriched nauplii reached DHA and EPA concentrations of 23.8 and 18.7 mg/g dry weight (dwt), respectively, while in Cojinoba larvae they were 18.4 and 19.7 mg/g dwt. Control emulsion (CE)‐enriched nauplii exhibited lower DHA and EPA (6.1 and 7.7 mg/g dwt), while only DHA decreased in the control larvae (12.6 mg/g dwt). Vitamin E was higher in EE‐enriched nauplii (29.2 mg/100 g dwt) than in the control (8.4 mg/100 g dwt). Larvae fed EE‐enriched Artemia exhibited 8% increase in survival and 19% in growth compared with the control. Astaxanthin was detected only in larvae fed EE‐enriched nauplii. The tumor necrosis factor‐α concentration was not significantly different between larvae fed EE‐ and CE‐enriched nauplii. EE looks promising as an Artemia enrichment and experimental diet to assess palm ruff larval requirements, and has a positive impact on fish larvae performance.     

饲料中DHA/EPA值对星斑川鲽幼鱼生长、体组成及血清生理指标的影响

为研究饲料DHA/EPA值对星斑川鲽幼鱼生长、体组成和血液生理指标的影响,实验配制等氮、等能的5种不同DHA/EPA值(0.64、0.97、1.18、1.59和1.91)的饲料,每个比值设3个重复,饲养周期56 d。结果显示:(1)随着饲料DHA/EPA值的升高,星斑川鲽幼鱼增重率、饲料效率、蛋白质效率均呈先上升后下降的趋势(P0.05)。当饲料DHA/EPA值为0.97~1.59时实验鱼增重最快,饲料效率最高。蛋白质效率则在DHA/EPA值为0.97~1.18时达到最高。蛋白质沉积率(protein retention efficiency,PRE)与饲料DHA/EPA值呈显著二次回归关系(y=-1.589 5x2+2.858 3x+45.184;R2=0.910 8,x=饲料DHA/EPA值,y=PRE),当饲料DHA/EPA值大于0.90时呈下降趋势。肝体比呈先下降后小幅回升的趋势(P0.05),在饲料DHA/EPA值为1.18时达到最低,为2.85%,脾脏指数呈显著上升趋势(P0.05),于饲料DHA/EPA值为1.59组最高(0.12%);(2)肝脏粗脂肪含量随饲料DHA/EPA值的增加呈明显下降趋势(P0.05),且在饲料DHA/EPA值为1.18时降到最低,为8.60%,而后又显著上升,但仍显著低于饲料DHA/EPA值为0.64时的水平(13.44%)。二次回归分析(y=5.199 6x2-15.652x+20.866;R2=0.634 8,x=饲料DHA/EPA值,y=肝脏脂肪含量)显示,当饲料中DHA/EPA值为1.51时肝脏脂肪含量最低。脂肪酸分析结果显示,随着饲料DHA/EPA值的升高,肝脏及肌肉中EPA含量均呈线性下降趋势(P0.05),而DHA含量及DHA/EPA均呈直线上升趋势(P0.05)。肝脏和肌肉组织n-3 HUFA总量不受饲料处理的影响(P0.05);(3)血清总蛋白、球蛋白含量在饲料DHA/EPA值为1.59时显著高于其他各组(P0.05),白蛋白在饲料DHA/EPA值为0.64、0.97和1.59水平最高。溶菌酶(LSZ)活性在饲料DHA/EPA值为1.18时达到峰值(P0.05),为2.76μg/mL。谷丙转氨酶(ALT)活性在饲料DHA/EPA值1.91时无显著变化,而当饲料DHA/EPA1.18时,血清谷草转氨酶(AST)活性提高了65%左右。研究表明,在本实验条件下,以增重率为参考指标,采用二次回归(y=-31.066x2+77.26x+76.541;R2=0.957 4,x=饲料DHA/EPA值,y=增重率)分析可得,当饲料脂肪水平为8.3%,n-3 HUFA含量为0.74%时,星斑川鲽幼鱼[初始体质量(31.70±0.12)g]对DHA/EPA值的最适需要量应为1.24。     

Pharmacokinetics and bioavailabilities of 14C-keto-carotenoids, astaxanthin and canthaxanthin, in rainbow trout, Oncorhynchus mykiss

The pharmacokinetics and bioavailabilities of ¹⁴C‐astaxanthin and ¹⁴C‐canthaxanthin were studied in the blood of rainbow trout following intra‐arterial (i.a.) and oral (p.o.) administration. Sixteen months old 1 kg trout were cannulated in the dorsal aorta. [6,7,6′,7′‐¹⁴C]‐keto‐carotenoids were administered i.a. and p.o. at a dose of 573.5 kBq kg^?1 fish body weight for astaxanthin and 836.2 kBq kg^?1 fish body weight for canthaxanthin. After i.a. distribution, total body clearance (Cl_tot) was 17.30±20.29 mL kg^?1 of fish h^?1 for ¹⁴C‐canthaxanthin and 3.30±1.50 mL kg^?1 of fish h^?1 for ¹⁴C‐astaxanthin. The volume of distribution at steady‐state (V_ss) was 208.32±124.79 mL kg^?1 of fish and 71.84±64.15 mL kg^?1 of fish for ¹⁴C‐canthaxanthin and ¹⁴C‐astaxanthin respectively. Less than 0.4% of the administered radioactivity was recovered in urine. Radioactivity (expressed as percent of the dose) excreted in the bile of fish that received ¹⁴C‐canthaxanthin by i.a. route was 20‐fold higher than that observed for fish treated p.o. This ratio was lower for ¹⁴C‐astaxanthin (7.6‐fold). The mean keto‐carotenoid bioavailabilities calculated were 10–15% for both compounds. Findings suggest one daily astaxanthin application is preferable, while 12‐h time intervals between applications are preferable for canthaxanthin.     

Effect of high dietary intakes of vitamin E and n-3 HUFA on immune responses and resistance to Edwardsiella tarda challenge in Japanese flounder (Paralichthys olivaceus, Temminck and Schlegel)

A feeding experiment was conducted to investigate the effects of high dietary intake of vitamin E (supplied as dl ‐α‐tocopheryl acetate) and n‐3 highly unsaturated fatty acid (n‐3 HUFA) on the non‐specific immune response and disease resistance in Japanese flounder Paralichthys olivaceus. Nine practical diets were formulated to contain one of three levels of vitamin E namely, 0, 80 or 200 mg kg^?1 (the total α‐tocopherol contents in the diets were 21, 97 and 213 mg kg^?1 based on analysis), and at each vitamin E level with one of three n‐3 HUFA levels i.e. 0.5%, 1.5% or 2.0%. Each diet was randomly assigned to triplicate groups of Japanese flounder (initial body weight: 40.5±1.0 g, mean±SD) in a re‐circulation rearing system. Fish were fed twice daily to apparent satiation at 07:00 and 18:00 hours for 12 weeks. During the experimental period, water temperature was maintained at 18±1°C, salinity 31–35 g L^?1, and pH 7.8–8.2. Dissolved oxygen was not less than 6 mg L^?1, and there were negligible levels of free ammonia and nitrite. The results showed that the increase in dietary n‐3 HUFA from 0.5% to 1.0% significantly decreased muscle α‐tocopherol contents in fish‐fed diets with 21 and 97 mg α‐tocopherol kg^?1 diet (P<0.05). In 1.0% HUFA groups, alternative complement pathway activity (ACH₅₀) of fish fed the diet containing the 213 mg α‐tocopherol kg^?1 diet was significantly higher than noted for fish fed the diet containing 97 mg α‐tocopherol kg^?1 diet (P<0.05). Fish fed the diet with 213 mg α‐tocopherol kg^?1 and 2.0% n‐3 HUFA had the highest lysozyme activity (131.7 U mL^?1) among all the dietary treatments. Fish fed the diets containing 97 and 213 mg α‐tocopherol kg^?1 with 1.0% n‐3 HUFA had significantly higher respiratory burst activity than those fed the diets containing 21 mg α‐tocopherol kg^?1 with 0.5 and 1.0% n‐3 HUFA (P<0.05). In the disease resistance experiment, high intake of dietary vitamin E with 213 mg α‐tocopherol kg^?1 significantly decreased cumulative mortality and delayed the days to first mortality after a 7‐day Edwardsiella tarda challenge (P<0.05). In addition, under the experimental conditions, dietary vitamin E and n‐3 HUFA had a synergistic effect on the non‐specific immune responses and disease resistance in Japanese flounder (P<0.05).     

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.     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.     

Molecular adaptations of glucose and lipid metabolism to different levels of dietary carbohydrates in juvenile Japanese flounder Paralichthys olivaceus

Japanese flounder juveniles (initial body weight: 7.12 ± 0.02 g) were fed three diets containing 0, 120 and 200 g/kg of carbohydrates (C0, C120 and C200, respectively) for 10 weeks. Results showed that higher dietary carbohydrate intake enabled further deposition of glycogen and lipids in liver and muscle. The mRNA levels of glucokinase (gck), phosphofructokinase (pfkl) and hexokinase (HK) activity involved in glycolysis were significantly up‐regulated in C120 (p < .05) but showed no further up‐regulation except for gck in C200. Besides, the gluconeogenic phosphoenolpyruvate carboxykinase 1 (pck1) mRNA levels were down‐regulated significantly in fish fed the C120 (p < .05). However, further increase in dietary carbohydrate levels increased the mRNA levels and activities of enzymes involved in gluconeogenesis and lipolysis (p < .05). Additionally, plasma glucose remained unchanged in C120 (p > .05) but significantly increased in C200 group (p < .05). In conclusion, Japanese flounder was able to use carbohydrates efficiently through regulation of glucose and lipid metabolism when dietary carbohydrate was not higher than 120 g/kg, while 200 g/kg of dietary carbohydrate caused the deregulation of glucose homoeostasis.     

维生素C对大鲵消化系统结构和功能的影响

本试验在大鲵基础饲料中添加不同梯度的维生素C,探究其对机体消化系统各器官结构和功能的影响。[方法]以鱼粉、鸡肉粉等为蛋白源,鱼油为脂肪源配制大鲵基础饲料,基础饲料中分别添加0 mg/kg(D1)、150mg/kg(D2)、300mg/kg(D3)、450 mg/kg(D4)、600mg/kg(D5)和750mg/kg(D6)的维生素C(维生素C磷酸酯,35%含量),配制成6种等氮等脂的试验饲料,饲喂初始体质量为(34.14±0.15)g 的试验幼鲵。[结果]结果表明：随维生素C添加量的增加,幼鲵胃蛋白酶、H⁺-K⁺-ATP酶活性均在D3组达到最高,通过对胃切片肌层厚度、绒毛高度等统计,添加300mg/kg维生素C比添加0、600mg/kg更有利于大鲵胃部发育(P＜0.05)；肠道糜蛋白酶、脂肪酶及Na⁺-K⁺-ATP酶活力均呈先升后趋于平稳趋势,在D4组达到最高(P＜0.05),对肠道淀粉酶无显著性影响(P＞0.05),通过对肠道切片肌层厚度、绒毛高度统计,添加300mg/kg维生素C比添加0、600mg/kg更有利于大鲵肠道发育；胰腺糜蛋白酶及脂肪酶活力均呈先升后趋于平稳趋势,分别在D3、D4组达到最大(P＜0.05),对胰腺淀粉酶无显著性影响(P＞0.05)；肝脏MDA、AST和ALT活性均呈先降后升趋势,并均在D3组达到最低,而CAT、T-SOD、ACP及AKP则呈先升后降的趋势,CAT、T-SOD、ACP在D4组达最高峰,AKP在D3组达最高,通过对肝脏切片发现,添加300mg/kg维生素C相较于添加0、600mg/kg炎性细胞浸润现象明显减少,肝巨噬细胞数量明显增加。[结论] 综上所述,适量的维生素C可有效的改善大鲵消化道的结构,提高消化酶活性,增强肝脏抗氧化能力,保护肝脏健康。     

Effect of dietary fish protein hydrolysate on growth,feed utilization and IGF‐I levels of Japanese flounder (Paralichthys olivaceus)

This study was conducted to investigate the effect of fish protein hydrolysate on growth performance, insulin‐like growth factor I (IGF‐I) levels and the expression levels of liver IGF‐I mRNA in juvenile Japanese flounder (Paralichthys olivaceus). Fish hydrolysate was produced by enzymatic (alcalase and flavourzyme) treatment and size‐fractionated by ultrafiltration. The permeate after ultrafiltration (UF) and the non‐ultrafiltered fish hydrolysate were tested as feed ingredients using high plant protein diets. Fish meal was used in the control diet (FM). The feeding trial lasted for 60 days, and fish fed with 37 g kg^?1 UF showed the best growth, feed efficiency, digestibility and protein utilization. Plasma IGF‐I level was examined with radioimmunoassay, and the expression levels of liver IGF‐I mRNA were evaluated using real‐time PCR normalized against the 18S rRNA gene. Plasma IGF‐I levels were significantly increased by inclusion of fish protein hydrolysate. Liver IGF‐I mRNA expression was significantly higher in fish fed with 37 g kg^?1 UF diet than fish fed with control diet. The results indicated that small molecular weight compounds from fish protein hydrolysate showed a positive effect on growth and feed utilization in juvenile Japanese flounder. Dietary fish protein hydrolysate could improve plasma IGF‐I levels and liver IGF‐I mRNA expression in Japanese flounder.     

Hg2+对克隆牙鲆和普通牙鲆的急性毒性

以相同规格的牙鲆Paralichthys olivaceus普通群体为对照,采用静水生物测试法开展了Hg2+对牙鲆克隆群体的急性毒性实验,实验所用幼鱼为130日龄,体长为(12.38±1.13)cm,体质量为(17.97±5.79)g。实验期间,水温为(21±1.2)℃,pH为8.1-8.3,盐度为31.0,硬度6200 mg/L,连续充气保证溶氧充足。研究表明,实验初始阶段,高浓度组克隆牙鲆在容器内躁动不安,上升到水面沿池壁缓慢转圈游动,一段时间后安静伏在池底;而普通牙鲆则仅个别鱼在水面游动,多数静止在池底不动。随中毒程度加深,用玻璃棒触碰普通牙鲆时,其反应剧烈,在水中侧翻、打转、急速游动或上下窜动,而克隆牙鲆则仅沿池底缓慢游动。最终克隆牙鲆和普通牙鲆均身体僵直,失去呼吸能力死亡。Hg2+对克隆牙鲆的24、48、96 h的半致死质量浓度分别为1.50、0.86、0.84 mg/L,而对普通牙鲆的半致死质量浓度则分别为1.93、1.04、0.90 mg/L。结果表明,克隆牙鲆对Hg2+的耐受性弱于普通牙鲆,对Hg2+更加敏感。从死亡过程看,各处理组的克隆牙鲆从出现死亡个体开始,死亡时间集中在8 h内,具有同步性;普通牙鲆则在实验期间陆续出现死亡,持续时间较长,同步性较差。     

Pharmacokinetics of Dietary 13C-Labeled Docosahexaenoic Acid and Docosapentaenoic Acid in Red Sea Bream Chrysophrys major

The objectives of this study were to investigate: 1) the pharmacokinetics of dietary docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA) using ¹³C-labeled fatty acids; 2) the interorgan transport of DHA in the red sea bream by monitoring the DHA level of several organs; and 3) the relationship between the plasma DHA level and optimum dietary DHA level in the plasma of the red sea bream Chrysophrys major . For this purpose, a mixture of 38.5% of [¹³C]DHA, 8.5% of [¹³C]DPA, and 4.2% of [¹³C]palmitic acid were given to the red sea bream at dose level of 8.0, 16.0, and 47.9 mg/kg by a single oral administration. For [¹³C]DHA, the maximum plasma concentration (t_max) occurred at 2.00–3.00 h after the oral administration. The peak plasma concentration (C_max) and the area under the plasma concentration-time curve to 24 h (AUC_0-24 for [¹³C]DHA level linearly increased with respect to dosage. [¹³C]DHA appeared in each organ (plasma, erythrocyte and the fat body of the orbit, liver, intestine, skin, brain, heart and muscle) at 0.5 h and was observed until 24 h. From the values determined for the pharmacokinetic parameters, the range of the effective plasma DHA level for normal growth of the red sea bream was suggested to be between 21.0 and 40.3 μg/mL. For [¹³C]DPA, the AUC_0-24 and C_max values also linearly increased with the dosage, but t_max did not depend on it.     

High‐throughput Cryopreservation of Sperm from Sex‐reversed Southern Flounder,Paralichthys lethostigma

The Southern flounder, Paralichthys lethostigma, is a valuable aquaculture fish with established markets in the USA. All‐female production in this species is an important technology for aquaculture because the females usually have body sizes twice those of males at the same age, and sex‐reversed males (genotypic XX neomales) are used for all‐female production by crossing with genetically normal females. However, sperm volume from the neomales is usually small (<0.5 mL) and limits their application for all‐female fish production. Cryopreservation of sperm from these sex‐reversed neomales will provide access on demand with increased efficiency to extend the application of neomales. The goal of this study was to develop a protocol for cryopreservation of sperm from the Southern flounder by using an automated high‐throughput processing system. The objectives were to: (1) determine the effect of osmolality on activation of sperm motility; (2) evaluate the effect of extender solutions on sperm motility capacity; (3) evaluate the acute toxicity of cryoprotectants (dimethyl sulfoxide [DMSO], propylene glycol, and polyethylene glycol) on sperm motility, and (4) estimate the effect of cooling rate on sperm cryopreservation and post‐thaw fertilization. Sperm motility was activated when osmolality was 400 mOsmol/kg or higher. Of the three extender buffers tested, HEPES4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES) at 300 mOsmol/kg resulted in better protection for sperm motility than did Hanks' balanced salt solution and Mounib solution at 300 mOsmol/kg during 7 d of refrigerated storage. After 30 min equilibration with the cryoprotectant of 15% DMSO, sperm motility was 24 ± 21% (fresh sperm motility without any cryoprotectants was 42%). After cooling at a rate of 20 C/min, post‐thaw sperm motility was 8 ± 5% and fertilization was 63 ± 40% evaluated at the 32–64 cell stage (5 × 10⁵ sperm per egg). Overall, a protocol was developed for sperm cryopreservation in the Southern flounder with high‐throughput processing, which provides a tool to preserve the valuable genetic resources from neomale flounders, and enables germplasm repository development for the Southern flounder.     

Low pathogenicity of flounder iridovirus (FLIV) and the absence of cross‐protection between FLIV and rock bream iridovirus

The genus Megalocytivirus is known to infect a wide range of cultured marine fish. In this study, we examined the pathogenicity of FLIV (Megalocytivirus from olive flounder, genotype III) and RBIV (Megalocytivirus from rock bream, genotype I) to their homologous and heterologous host species. Olive flounder (7.5 ± 1.3 cm) injected with FLIV [major capsid protein (MCP) gene copies, 6.8 × 10³–6.5 × 10⁶/fish] at 24 °C did not die until 90 days post‐infection (dpi). The average virus replication in the spleen peaked (1.27 × 10⁶/fish) at 20 dpi. Rock bream (6.5 ± 1.5 cm) injected with FLIV (8.8 × 10⁵ and 6.5 × 10⁶/fish of MCP copies) showed no mortality until 50 dpi. The rock bream that survived after FLIV infection were rechallenged with RBIV at 50 dpi had 100% mortality, showing that there is no cross‐protection between FLIV and RBIV. Temperature shifting (26 °C and 20 °C at 12 h intervals) did not cause FLIV‐specific mortality into olive flounder, but higher virus copies were observed in the fish exposed to higher stocking density. This study demonstrates that FLIV and RBIV have different antigenic and pathogenic characteristics and that FLIV has low pathogenicity to olive flounder.     

Induction of ovulation in ornamental common carp (Koi, Cyprinus carpio L.) using LHRHa ([d-Ser(tBu)6, Pro9-NEt]-LHRH) combined with haloperidol and carp pituitary extract

The effects of a single injection of mammalian superactive analogue [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa (20 μg/ kg^?1) combined with the dopamine antagonist, haloperidol (HAL, 0.5 mg kg^?1), for induction of ovulation in the koi carp broodstocks were determined under routine hatchery conditions. The results were compared with classic carp pituitary extract (CPE, double injection) application (water temperature 22 °C). Physiological saline (0.7% NaCl)‐injected fish were used as a control group and no ovulation occurred in this group. The spawning ratio was high in the LHRHa+HAL treatment group and in the CPE treatment group (6/7 and 5/7 respectively). The latency period was 14–16 h in the LHRHa+HAL treatment group and 12–14 h in the CPE treatment group (after the second injection). There was no difference between the two ovulating groups with respect to the spawning index (the weight of eggs as a percentage of female body weight) and fertilization rate of eggs (P>0.05). As a result, ovulation can be induced successfully in koi carp broodstocks with 20 μg kg^?1 [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa+0.5 mg kg^?1 HAL treatment in a single injection without decreasing the egg quality. Application of this combination can be useful for hatchery and broodstock management in koi carp culture.     

Nutritional comparison in muscle of wild,pond and factory cultured Japanese flounder (Paralichthys olivaceus) adults

Muscle proximate composition, amino acid profiles, fatty acid compositions and selected minerals of wild, pond and factory cultured Japanese flounder adults were compared in this study to elucidate their nutritive values and dietary nutrition requirements. Wild and pond cultured Japanese flounder flesh had higher crude proteins, but crude lipid contents of factory cultured fish were 3.8~4.0 fold greater. Major amino acids in Japanese flounder were aspartic acid, glutamic acid and lysine. Wild fish had higher levels of total amino acids, essential amino acids, nonessential amino acids and half‐essential amino acids and the contents in pond cultured samples were similar. Arachidonic acids (ARA), eicosapentaenoic acids (EPA), docosahexaenoic acids (DHA), polyunsaturated fatty acids (PUFA), DHA + EPA and n–3 PUFA in wild Japanese flounder were significantly higher than that in factory cultured fish, whereas fatty acid levels were close between pond cultured and wild fish. Japanese flounder were rich in zinc, iron and selenium, but poor in copper, chromium and nickel. Results indicated wild Japanese flounder had higher nutritional value and better meat quality, but the nutritional compositions of pond cultured fish were close to wild Japanese flounder, which suggested that pond culture of Japanese flounder offers broad application prospects.     

Combined Effects of Turbulence and Salinity on Growth, Survival, and Whole-body Osmolality of Larval Southern Flounder

The southern flounder (Paralichthys lethostigma) is a commercially important marine flatfish from the southeastern Atlantic and Gulf Coasts of the USA and an attractive candidate for aquaculture. Hatchery methods are relatively well developed for southern flounder; however, knowledge of the optimum environmental conditions for culturing the larval stages is needed to make these technologies more cost effective. The objectives of this study were to determine the effects of water turbulence (as controlled by varying rates of diffused aeration) on growth, survival, and whole‐body osmolality of larval southern flounder from hatching through day 16 posthatching (d16ph). Embryos were stocked into black 15‐L cylindrical tanks under four turbulence levels (20, 90, 170, and 250 mL/min of diffused aeration) and two salinities (24 and 35 ppt) in a 4 × 2 factorial design. Larvae were provided with enriched s‐type rotifers from d2ph at a density of 10 individuals/mL. Temperature was 19 C, light intensity was 390 lx, and photoperiod was 18 L:6 D. Significant (P < 0.05) effects of turbulence on growth (notochord length [NL], wet weight, and dry weight) were observed. On d16ph, NL (μm) increased with decreasing turbulence level and was significantly greater at 20 mL/min (64.2) and 90 mL/min (58.2) than at 170 mL/min (56.3) and 250 mL/min (57.2). Survival declined primarily during the prefeeding and first‐feeding stages from d0 to d8ph, then stabilized from d8 to d16ph. In contrast to growth trends, survival (%) on d16ph increased with increasing turbulence levels and was significantly greater at 170 mL/min (57.9) and 250 mL/min (54.0) than at 20 and 90 mL/min (21.4 and 26.2, respectively). Mean rotifer concentrations (individuals/mL) at 24 h postfeeding were significantly higher (P < 0.05) in the low‐turbulence treatments of 20 mL/min (4.48) and 90 mL/min (4.23) than in the high‐turbulence treatments of 170 and 250 mL/min (2.28 and 2.45, respectively). Under both salinities, larval whole‐body osmolality (mOsm/kg) increased with increasing turbulence levels and was significantly higher at 250 mL/min (427) than at 20 mL/min (381), indicating osmoregulatory stress at the higher turbulence levels. On d14ph, larvae in all treatments were positively buoyant in 35 ppt and negatively buoyant in 24 ppt. Results showed that growth of southern flounder larvae in 15‐L tanks was maximized under low turbulence levels of 20 and 90 mL/min, while survival was maximized at high turbulence levels of 170 and 250 mL/min. The data suggested that, in prefeeding‐ and early‐feeding‐stage larvae (which have weak swimming ability), higher turbulence levels improved buoyancy and prevented sinking. In feeding‐stage larvae (which are relatively strong swimmers), higher turbulence levels caused excessive swimming, osmoregulatory stress, and slower growth. Based on these results, we recommend that turbulence levels be maintained relatively high during prefeeding (yolk sac) and first‐feeding stages to maintain buoyancy and survival and then decreased for mid‐ to late‐feeding‐ and premetamorphic stage larvae to optimize prey encounters and feeding efficiency.     

Growth and Feed Utilization of Captive Wild Black Sea Bass Centropristis striata at Four Different Densities in a Recirculating Tank System

A study to determine the effects of four stocking densities on growth and feed utilization of wild‐caught black sea bass Centropristis striata was conducted in a pilot‐scale recirculating tank system. The outdoor system consisted of 12 insulated fiberglass tanks (dia. = 1.85 m; vol. = 2.17 m³) supported by biological filters, UV sterilizers, and heat pumps. Subadults (N= 525; ×± SD = 249 ± 16.8 g) were stocked at densities of 4.6 fish/m³ (1.18 kg/m³), 16 fish/ m³ (3.91 kg/m³), 25.3 fish/m³ (6.83 kg/m³), and 36 fish/m³ (7.95 kg1m³), with three replicate tanks per treatment. Fish were grown under 35 ppt salinity, 21‐25 C, and under ambient photoperiod conditions. A commercial flounder diet containing 50% protein and 12% lipid was hand‐fed twice daily to satiation for 201 d. Mean (range) total ammonia‐nitrogen, 0.61 (0‐2.1) mg/L, nitrite‐nitrogen, 0.77 (0.04‐3.6) mg/L, and nitrate‐nitrogen 40.1 (0‐306) mg/L were significantly higher (P < 0.0001) in the 25.3 and 36 fish/m³ treatments than in the 4.6 and 16 fish/m³ treatments [0.19 (0.05‐0.5), 0.1 (0.24‐0.63), and 11.9 (1.3‐82.2) mg/L, respectively]. However, there were no significant differences (P > 0.05) in growth (RGR = 196.8‐243.1%; DWG = 2.55‐2.83 g/d; and SGR = 0.55‐0.61%/d), coefficient of variation of body weight (C_wtV., = 0.24‐0.25), condition factor (K = 2.2‐2.4), feed consumption (FC = 1.45‐1.65%/d), and feed conversion ratio (FCR = 1.45‐1.52) among stocking densities. Final biomass densities on day 201 reached 3.48, 12.0, 21.1, and 27.2 kg/m³ at stocking densities of 4.6, 16, 25.3, and 36 fish/m³, respectively. Survival (83.8‐99.1%) did not differ among treatments. Apparent net protein retention (ANPR) was significantly higher (P < 0.005) for fish stocked at the lower densities of 4.6 and 16 fish/m3 (22.5‐23.7%) than for those stocked at 25.3 and 36 fish/m³ (21‐20.1%). There were no significant differences (P > 0.05) in apparent net energy retention (ANER = 55.9‐59.1 %) among stocking densities. Final whole body protein (15.3‐16.3%) and lipid (23.1‐26.4%) levels did not differ significantly (P > 0.05) among treatments. The results demonstrated that growth, survival, and feed utilization were not impaired under stocking densities ranging from 4.6‐36 fish/m³ (3.48‐27.2 kg/m3), despite a slight reduction in water quality at the higher densities. In addition, growth variation and final whole body protein and lipid levels were not influenced by these densities. The results suggest that black sea bass are tolerant of crowding and moderate variations in water quality during intensive culture in recirculating tank systems and that higher stocking densities are possible.     

Dietary ratios of n‐3/n‐6 fatty acids do not affect growth of Nile tilapia at optimal temperatures (28°C) nor at temperatures that simulate the onset of winter (22°C)

Nile tilapia (Oreochromis niloticus) juveniles were fed diets containing 13 g/kg total polyunsaturated fatty acids (PUFAs) at different n‐3/n‐6 dietary ratios (0.2, 0.5, 0.8, 1.3 and 2.9) for 56 days, at 28°C. Subsequently, fish were submitted to a winter‐onset simulation (22°C) for 33 days. PUFA n‐3/n‐6 dietary ratios did not affect fish growth at either temperature. At 28°C, tilapia body fat composition increased with decreasing dietary PUFA n‐3/n‐6. Winter‐onset simulation significantly changed feed intake. The lowest dietary n‐3/n‐6 ratio resulted in the highest feed intake. At both temperatures, body concentrations of α‐linolenic acid, docosahexaenoic acid, eicosatrienoic acid and docosapentaenoic acid decreased as dietary n‐3/n‐6 decreased. Body concentrations of eicosapentaenoic acid (EPA, 20:5 n‐3) increased with decreasing concentrations of dietary EPA. The n‐6 fatty acids with the highest concentrations in tilapia bodies were linoleic acid and arachidonic acid (ARA, 20:4 n‐6). At 28°C, SREBP1 gene expression was upregulated in tilapia fed the lowest n‐3/n‐6 diet compared to tilapia fed the highest n‐3/n‐6 ratio diet. Our results demonstrate that a dietary PUFA of 13 g/kg, regardless of the n‐3/n‐6 ratio, can promote weight gains of 2.65 g/fish per day at 28°C and 2.35 g/fish per day at 22°C.