真鲷与黑鲷杂交繁育技术研究

通过对真鲷黑鲷杂交繁育,培育出一种能结合父母本性状,具有生长速度快、抗逆性强的杂交子一代。2006年3月从海区选购真鲷（Pagrosomusmajor）、黑鲷（Spraus macrocephalus）亲鱼,通过强化培育,性腺促熟,于4月进行干法人工授精。通过对真鲷♀×黑鲷8及黑鲷♀×真鲷♂获得杂交受精卵,结果真鲷♀×黑鲷♂受精率为87．4％,孵化率为85％,出苗率14．28％;黑鲷♀×真鲷♂受精率为92．1％,孵化率为90％,出苗率7．2％,该批次共获得正交反交杂交鱼苗32000尾。从杂交实际结果看,不管正交还是反交都比自交的结果差,说明它们之间存在生殖隔离,杂交繁育难度相当大,要达到规模生产。需要继续研究、试验。     

日粮水平和饵料种类对黑鲷能量收支的影响

采用室内流水实验法研究了摄食水平和饵料种类对黑鲷能量收支各组分和模式的影响。结果表明，黑鲷的生长量，总呼吸代谢量，排泄量及生态转换效率均随日粮水平升高呈增长趋势，二者之间的定量关系可分别用对数曲线或直线加以定量描述；不同生物饵料能造成黑鲷摄食，生长，排泄和总呼吸代水平的显著差异，但却不能改变以比能值为单位的生态转换效率。黑鲷的呼能量收支分配率随摄食水平不同有较显著差异；代谢能分配率和排泄能分配率随摄食水平增大呈U型变化趋势，而生长能分配率却恰恰相反。除摄食高蛋白饵料时引起排泄能分配率增加外，摄食不同饵料对黑铜的能量分配模式影响不十分显著。     

土霉素在黑鲷体内的药物代谢动力学研究

首次报道了黑鲷口服土霉素的药物代谢动力学特征，用高效液相色谱法测定组织中的药物含量，药物在肌肉，血液，肝脏中的平均回归率分别为８５.61%,85.38%,82.005,该方法的检测限可达0.01μg/g，黑鲷１次口服剂量为７５mg/kg的土壤素后，其血液药物浓度－时间数据符合一室开放动力学模型，吸收速率常数（ka)为０.296/h，达峰时间（Ｔamx)为１０.635h,峰浓度（Ｃmax)为１.398μg/ml, 分布半衰期（Ｔ１／２a）为２.339h ,消除半衰期(T1/20β）为46.663h，药时曲线下面积（ＡＵＣ）为１１0.25mg/L.h，黑绸口服药物０.5h后在血液，肥肉，肝脏，肾脏４种组织中就可以检测到药物的存在，药物在１６h的采样点浓度达最高，分别为1.68μg/ml,1.68,2.52,6.77μg/g。     

大头鳕TNFSF6基因的结构分析及其在发育早期和病毒暴发时的表达水平

为揭示大头鳕TNFSF6基因的基本结构与功能及其在大头鳕发育和神经坏死病毒(Pacific cod nervous necrosis virus, PCNNV)暴发时的响应机制,本研究通过基因克隆获得TNFSF6 cDNA开放阅读框序列,并对序列进行生物信息学分析,运用相对荧光定量PCR(qRT-PCR)方法对TNFSF6在不同组织、孵化后不同日龄仔鱼和PCNNV感染前后仔稚鱼中的表达水平进行检测。结果显示,大头鳕TNFSF6 cDNA长1 388 bp,5′UTR占315bp,3′UTR占500 bp,ORF全长573 bp,编码190个氨基酸。qRT-PCR结果显示,TNFSF6在各组织均有表达,但在脾脏和鳃组织中的表达量较高;大头鳕孵化后15、20、37和40 d TNFSF6基因的转录水平分别是其在5 d转录水平的0.28、0.15、0.12和0.13倍。在24 d和46 d PCNNV暴发时,病鱼TNFSF6基因的转录水平高于对照组;在77 d PCNNV暴发时,病鱼TNFSF6转录水平则显著低于对照组。研究表明,TNFSF6基因在大头鳕发育早期和仔鱼暴发PCNNV时发挥了重要的作用。     

鳕碎肉酶解物清除羟自由基作用研究

通过测定鳕碎肉酶解物对Fenton体系产生的羟自由基的清除效果,从海洋蛋白酶YS-894、海洋蛋白酶YS-80、胃蛋白酶和木瓜蛋白酶中,筛选出海洋蛋白酶YS-894作为酶解鳕碎肉制备具有较高清除羟自由基活性酶解物的水解酶;用正交试验L9(34)对该酶的水解条件进行优化,并确定了水解的最佳肉水比。最终工艺条件为40℃、酶解45 m in、pH 9.0、酶解质量分数0.25%、底物∶水=1∶3的条件下进行水解,水解物对羟自由基的清除效果较好,清除率为68.88%。     

饲喂频率对黑鲷生长及体生化成分的影响

选取同一种饵料设置4个饲喂频率水平(分别为1次/d、2次/d、4次/d、6次/d),采用静水连续充气养殖系统,在盐度为26.0~27.0、温度为26.8~24.5℃的条件下对黑鲷成鱼和鱼种进行为期40d的生长实验。结果表明:(1)摄食率不受饲喂频率的显著影响,饲喂频率影响黑鲷的生长是由饲料转化率的改变引起的。(2)黑鲷成鱼和鱼种的水分含量随饲喂频率的增高而增加,黑鲷成鱼脂肪含量随饲喂频率的增高而下降,黑鲷鱼种脂肪含量随饲喂频率的增高而增加。(3)在本试验条件下,适宜饲喂频率随着黑鲷鱼体重的增加而提高,黑鲷鱼种的适宜饲喂频率为2次/d,黑鲷成鱼的适宜饲喂频率为4次/d。     

Effects of ontogeny, temperature, and light on vertical movements of larval Pacific cod (Gadus macrocephalus)

The role of behavior, especially vertical migration, is recognized as a critical component of realistic models of larval fish dispersion. Unfortunately, our understanding of these behaviors lags well behind our ability to construct three-dimensional flow-field models. Previous field studies of vertical behavior of larval Pacific cod ( Gadus macrocephalus ) were limited to small, preflexion stages (≤11 mm SL) in a narrow range of thermal conditions. To develop a more complete picture of larval behavior, we examined the effects of ontogeny, temperature, and light on vertical responses of larval Pacific cod in experimental columns. While eggs were strictly demersal, yolk-sac larvae displayed a strong surface orientation as early as 1 day post hatch (∼ 5 mm SL). Consistent with field observations, small preflexion larvae (<10 mm SL) showed no response to varying light levels. However, there was a direct effect of temperature on larval behavior: Pacific cod larvae exhibited a stronger surface orientation at 4°C than at 8°C. The behavior of larger, postflexion larvae (>15 mm SL) in experimental columns was consistent with a diel vertical migration and independent of water temperature: fish were more widely distributed in the column, and median positions were consistently deeper at higher light levels. These laboratory observations are combined with observations from discrete-depth (MOCNESS) sampling in the Gulf of Alaska to characterize the vertical distribution of larval Pacific cod and contrast ontogenetic patterns with walleye pollock ( Theragra chalcogramma ). The vertical movements of larval Pacific cod described here will be applied in the development of dispersal projections from Gulf of Alaska spawning grounds.     

An evaluation of the duration of efficacy of emamectin benzoate in the control of Caligus curtus Müller infestations in Atlantic cod, Gadus morhua L

The duration of efficacy of emamectin benzoate in the control of Caligus curtus infestations in Atlantic cod, Gadus morhua L., was studied following an administration of 50 μg kg^?1 for seven consecutive days. No lice were found on medicated fish when challenged 1 week (challenge 1) or 5 weeks (challenge 2) following termination of medication, whereas the mean abundance of lice among the unmedicated fish was 17.9 and 19.3 lice per fish in challenge 1 and 2, respectively. Muscle concentrations of 19.5 ± 8.2 ng g^?1 and 3.4 ± 0.9 ng g^?1, respectively, and skin concentrations of 23.1 ± 10.8 and 4.2 ± 1.0 ng g^?1, respectively, were found 27 and 55 days following the termination of medication. Tissue concentrations and the duration of efficacy indicate a dosing regime for emamectin in cod, similar to the regime used for Atlantic salmon, Salmo salar L.     

A method to determine protein digestibility of microdiets for larval and early juvenile fish

A method to evaluate protein quality using in vivo methods was developed for larval fish. FluoSpheres^® fluorescent microspheres (10 μm) were incorporated into two test diets, our standard zein microdiet (ZMD) and a microdiet with identical ingredients except for the replacement of high quality fish meal with the same product cooked for 24 h at 80 °C (ZMD-CF). Several trials were performed to design a reliable method to test digestibility using FluoSpheres^® as a marker. The developed in vivo technique was tested on 35 days posthatch (dph) larval Atlantic cod ( Gadus morhua L.) and two tropical fish species in the early juvenile stage. The method took into account loss of total protein to the faecal pellet and water column. Apparent digestibility of protein in larval cod fed ZMD was significantly higher than that of larvae fed ZMD-CF ( P  < 0.05). A growth study to validate differences between the two diets showed significant differences in growth and survival of larvae fed ZMD versus ZMD-CF ( P  < 0.05). Further validation of our results was indicated through the use of a pH-stat method using enzymes extracted from 35 dph larval cod guts. This novel technique will be advantageous for researchers to evaluate feed ingredients for larval marine fish and is adaptable to many different areas of larval fish nutrition.