滤食性贝类养殖活动对海域生态系统的影响及生态容量评估

滤食性贝类以水体中的浮游植物和有机碎屑为主要食物，养殖海域的初级生产力水平、水动力学特性等生态环境因子的差异，不仅直接影响养殖贝类的产量，而且也与贝类养殖活动对生态环境的压力密切相关。由于养殖的种类、密度、方式及养殖海域的特性不同，关于贝类对生态环境的影响往往有不同的结果。本文以我国北方大连獐子岛扇贝底播海区和荣成桑沟湾贝类筏式养殖区为研究对象，采用现场调查、室内受控实验及生态数值模型模拟方法，分析研究了滤食性贝类对海域生态系统的影响，对这两个海域贝类养殖的生态容量进行了初步的评价。 主要结果： 1. 獐子岛海域底播贝类养殖活动对该海域生态系统的影响较小。非参数统计—符号检验的结果显示，养殖区与非养殖区之间的溶解性无机氮、磷酸盐浓度、氮磷摩尔比及浮游植物群落结构没有统计学上的差异（p>0.05）。但是从变化的趋势上来看，贝类养殖活动对水域环境的某些参数有一定程度的影响。例如，獐子岛底播贝类养殖海域的溶解性无机氮以氨氮为主，可能与贝类的代泄活动有关；不论是叶绿素浓度，还是网采浮游植物的生物量都是贝类高密度养殖区<贝类低密度养殖区<非养殖区（7月份除外），这种趋势可能与贝类的摄食压力有关。 桑沟湾各环境指标表现出明显的区域性。除春季外，非养殖区的DIN浓度高于各养殖区。在春季和冬季，贝类区的磷酸盐浓度显著降低；而硅酸盐浓度在夏季和秋季显著增大。综合分析DIN、PO4-P及SiO3-Si三个参数的四季变化，海带区、贝藻区及贝类区发生显著性变异的概率分别为25%，42%和50%，贝类区的变异较大。浮游植物、小型浮游动物的生物多样性指数都是以非养殖区为最高，贝类区的多样性指数最低。尤其是浮游动物的丰度，贝类区显著低于非养殖区。 2. 利用挪威的MOM (Modelling-Ongrowing fish farms-Monitoring)评价系统，评价了桑沟湾长期大规模的贝藻筏式养殖活动对底质环境的压力。在桑沟湾设10个取样站位，共获得66个底泥样品。比较了MOM－B评价系统的3组参数的季节变化特性。结果显示，底质条件属于1级，说明桑沟湾贝藻长期大规模的养殖活动对底质环境的压力较低。结合桑沟湾的环境及养殖特点，分析了压力较低的原因。 3. 经计算，2006年中国海水养殖的贝类和藻类使用浅海生态系统的碳可达396万吨，并通过收获从海中移出至少136.9万吨的碳。从1995年至2006年，养殖大型藻类和贝类累计移出的碳分别约为365万吨和893万吨，总计达1258万吨。证明了浅海的贝类和藻类养殖活动直接或间接地使用了大量的海洋碳，提高了浅海生态系统吸收大气CO2的能力。 4. 采用模拟现场生物沉积法测定了虾夷扇贝的滤水率、摄食率等生理指标及其与贝类个体大小、水温的关系。虾夷扇贝单位个体的滤水率与组织干重的关系符合幂函数方程CR=a×DWb，b值在0.45～0.65范围内；水温对虾夷扇贝滤水率的影响极其显著(p<0.01)，温度（T）与滤水率（CR）呈抛物线的关系：CR=-0.0009T2+0.0188T-0.0306，水温为10℃时，虾夷扇贝的滤水率、摄食较大。 5. 采用模拟现场流水法测定了3种滤食性贝类的食物选择性。紫贻贝、长牡蛎及栉孔扇贝分别对直径4m, 6m 和 8m颗粒的保留效率达到最大值；对小颗粒（直径2m）的保留效率分别为17%, 19% 和 8％。栉孔扇贝对食物数量和质量浓度的变化相对敏感，随着数量浓度的增加，栉孔扇贝倾向于摄食较大的颗粒；随着颗粒食物质量浓度的增加，倾向于摄食较小的颗粒。 6. 獐子岛海域四个航次的调查结果显示，叶绿素浓度在1.23～2.85mg.m-3范围内，均值为1.78±0.57 mg.m-3；初级生产力的变化范围为30.4～117.0 mg C. m-2.d-1，平均值为76.6±41.9 mg C. m-2.d-1。通过虾夷扇贝生物量断面调查，获得了虾夷扇贝的壳高频率分布情况，7月份的众数值出现在100 mm，10月份壳高的众数值为80 mm。利用以上测定的虾夷扇贝的滤水率等基本生物学特性，结合虾夷扇贝的年产量、海域面积和有关的水文状况等数据，计算了滤水效率、摄食压力、调节比率3个食物限制性指标参数，全年的均值分别为0.048, 0.31和 0.16，都小于1，说明目前该海域虾夷扇贝的养殖量未达到养殖容量。 7. 利用STELLA软件，建立的桑沟湾贝藻养殖的数值模型，模拟了叶绿素a浓度及氮磷营养盐的周年变化情况，及其对贝类养殖生物量变化的响应。以叶绿素a浓度为指标，初步探讨了桑沟湾贝类的生态容量。

Effects of dietary cyanobacteria of two different sources on growth and recovery of hybrid tilapia (Oreochromis niloticus x O. aureus)

A 115 days feeding trial was conducted to evaluate the effect of dietary cyanobacteria on growth, microcystins (MCs) accumulation in hybrid tilapia (Oreochromis niloticus x O. aureus) and the recovery when the fish were free of cyanobacteria. Three experimental diets were formulated: the control (cyanobacteria free diet); one test diet with cyanobacteria from Lake Taihu (AMt 80.0 mu g MCs g(-1) diet) and one with cyanobacteria from Lake Dianchi (AMd, 410.0 rho g MCs g(-1) diet). Each diet was fed to fish for 60 days and then all fish were free of cyanobacteria for another 55 days. A significant increase in feeding rate (FR) was observed in fish fed AMd diet after a first 30-day exposure (1(st) EP), and in fish fed both AMt diet and AMd diet after a second 30-day exposure (2(nd) EP). Specific growth rates (SGR) of fish fed AMt diet and AMd diet were both obviously affected after the first 30-day exposure, but SGR was only significantly affected in fish fed AMt diet after the second 30-day exposure. After a 55-day recovery, there were no significant differences among diets in the indices mentioned above. Much higher concentrations of MCs were accumulated in tissues of all fish exposed to cyanobacteria. After the 55-day recovery, MC concentrations in fish tissues were significantly lower than those on day 60. (C) 2009 Elsevier Ltd. All rights reserved.

Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish

Short hairpin RNA (shRNA) directed by RNA polymerase III (Pol III) or Pol II promoter was shown to be capable of silencing gene expression, which should permit analyses of gene functions or as a potential therapeutic tool. However, the inhibitory effect of shRNA remains problematic in fish. We demonstrated that silencing efficiency by shRNA produced from the hybrid construct composed of the CMV enhancer or entire CMV promoter placed immediately upstream of a U6 promoter. When tested the exogenous gene, silencing of an enhanced green fluorescent protein (EGFP) target gene was 89.18 +/- 5.06% for CMVE-U6 promoter group and 88.26 +/- 6.46% for CMV-U6 promoter group. To test the hybrid promoters driving shRNA efficiency against an endogenous gene, we used shRNA against no tail (NTL) gene. When vectorized in the zebrafish, the hybrid constructs strongly repressed NTL gene expression. The NTL phenotype occupied 52.09 +/- 3.06% and 51.56 +/- 3.68% for CMVE-U6 promoter and CMV-U6 promoter groups, respectively. The NTL gene expression reduced 82.17 +/- 2.96% for CMVE-U6 promoter group and 83.06 +/- 2.38% for CMV-U6 promoter group. We concluded that the CMV enhancer or entire CMV promoter locating upstream of the U6-promoter could significantly improve inhibitory effect induced by the shRNA for both exogenous and endogenous genes compared with the CMV promoter or U6 promoter alone. In contrast, the two hybrid promoter constructs had similar effects on driving shRNA.

Simultaneous determination of microcystin-LR and its glutathione conjugate in fish tissues by liquid chromatography-tandem mass spectrometry

A sensitive and selective liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantitative determination of microcystin-LR (MC-LR) and its glutathione conjugate (MC-LR-GSH) in fish tissues. The analytes were extracted from fish liver and kidney using 0.01 M EDTA-Na-2-5% acetic acid, followed by a solid-phase extraction (SPE) on Oasis HLB and silica cartridges. High-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry, operating in selected reaction monitoring (SRM) mode, was used to quantify MC-LR and its glutathione conjugate in fish liver and kidney. Recoveries of analytes were assessed at three concentrations (0.2, 1.0, and 5 mu g g(-1) dry weight [DW]) and ranged from 91 to 103% for MC-LR, and from 65.0 to 75.7% for MC-LR-GSH. The assay was linear within the range from 0.02 to 5.0 mu g g(-1) DW, with a limit of quantification (LOQ) of 0.02 mu g g(-1) DW. The limit of detection (LOD) of the method was 0.007 mu g g(-1) DW in both fish liver and kidney. The overall precision was determined on three different days. The values for within- and between-day precision in liver and kidney were within 15%. This method was applied to the identification and quantification of MC-LR and its glutathione conjugate in liver and kidney of fish with acute exposure of MC-LR. (c) 2007 Elsevier B.V. All rights reserved.

Partial compensatory growth in hybrid tilapia Oreochromis mossambicus x O-niloticus following food deprivation

The capacity of hybrid tilapia Oreochromis mossambicus x O. niloticus [23.2 +/- 0.2 g (mean +/- SE)] to show compensatory growth was assessed in an 8-week experiment. Fish were deprived of feed for 1, 2 and 4 weeks, and then fed to satiation for 4 weeks; fish fed to satiation during the experiment served as control. Water temperature gradually declined from 28.1 to 25.5 degrees C throughout the experiment. Specific growth rate (SGR) decreased with progressive food deprivation. At the end of deprivation, body weight was lower in the deprived fish than in the control. Fish deprived for 4 weeks exhibited lower contents of lipids and energy in whole body, and higher moisture content and ratio of protein to energy (P/E) than those of the control; they also consumed feed faster than the control when normal feeding was resumed. All deprived fish showed higher food intake (FI) than that of the control during re-alimentation; however, enhanced SGR was only observed in the fish deprived for 4 weeks. There were no significant differences in digestibility of protein and energy, food efficiency (FE) or energy retention efficiency between the control and deprived fish. At the end of re-alimentation, deprived fish failed to catch up in body weight with the control, while content of moisture, lipids and energy, and P/E in whole body of the deprived fish did not significantly differ from that of the control. The results of the experiment revealed that the hybrid tilapia reared in freshwater showed partial capacity for compensatory growth following food deprivation of 4 weeks, and that growth compensation was due mainly to increased FI, rather than to improved FE.

Compensatory growth in hybrid tilapia, Oreochromis mossambicus x O-niloticus, reared in seawater

Hybrid tilapia weighing 4.34 +/- 0.03 g (mean +/- SE) were reared in seawater at 23.8 to 27.0 degrees C for 8 weeks. The control group was fed to satiation twice a day throughout the experiment. The other three groups were deprived of feed for 1, 2, and 4 weeks, respectively, and then fed to satiation during the refeeding period. At the end of the experiment, fish deprived for 1 week had similar body weights to the controls, whereas fish deprived for 2 and 4 weeks had significantly lower body weights than the controls. During the refeeding period, size-adjusted feed intakes and specific growth rates were significantly higher in deprived fish than in the controls, indicating some compensatory responses in these fish. Feed intake and growth rate upon refeeding were higher the longer the duration of deprivation. No significant differences were found in digestibility, feed efficiency or protein and energy retention efficiency between the deprived and control fish during refeeding, suggesting that hyperphagia was the mechanism responsible for increased growth rates during compensatory growth. During refeeding, relative gains in protein, lipid and ash, as proportions of total body weight gain, did not differ significantly among treatment groups. (C) 2000 Elsevier Science B.V. All rights reserved.