千里岩海域国家级水产种质资源保护区现状调查与评价

2015―2018年,对千里岩海域国家级水产种质资源保护区的海水、沉积物、海洋生物、保护生物资源概况进行调查和评价,并研究了主要保护对象刺参(Apostichopus japonicus)的遗传多样性。结果显示,保护区海水所测参数均符合评价标准,但氮磷比远高于Redfield比值,磷相对缺乏。潜在性富营养化评价显示,调查海域属Ⅰ级贫营养。沉积物均符合国家Ⅰ类质量标准。海洋生物多样性多处于较高水平(H¢≥2),但2015年8月小型浮游动物多样性偏低(H¢=0.93),2016年5月和2018年5月大型浮游动物多样性偏低(H¢=0.82,H¢=0.22)。综合评价认为,该保护区的海洋生态环境良好,满足保护区功能需求。保护区主要保护对象刺参的单倍型多样度指数为0.976,核苷酸多样度指数为0.033,遗传多样性较高,但资源量较保护地初建时下降;另一个保护对象皱纹盘鲍(Haliotis discus hannai)资源极少。需尽快采取强有力的措施开展保护区管理及资源恢复工作。     

海州湾大竹蛏种质资源保护区生态环境质量分析与评价

本研究根据2018年5、8月对海州湾大竹蛏(Solen grandis)国家级水产种质资源保护区表层海水营养盐、重金属、石油类及相关环境因子的调查数据,运用综合质量指数法对该保护区海水生态环境质量进行综合评价,并采用富营养化指数和有机污染指数从不同角度分析该保护区的海水环境质量。结果显示,除部分站位无机氮(DIN)含量略超标外,海州湾保护区其他监测因子均符合国家Ⅰ类海水水质标准。2018年海州湾保护区海水综合质量指数范围为0.31~0.39,属于尚清洁等级。N/P值均大于Redfield系数,且磷酸盐(PO4-P)含量较低,表明该海域营养盐结构为磷限制。富营养化指数范围为0.05~0.27,海水为贫营养状况。有机污染指数范围为-0.24~0.81,该水域有机污染程度较轻。与2008~2013年历史调查数据相比,近年来,海州湾保护区海水生态环境质量保持较好。     

天津大神堂牡蛎礁国家级海洋特别保护区水质分析

为了对天津大神堂牡蛎礁国家级海洋特别保护区进行水质环境分析,2012年4月、9月和2013年5月、9月分别对该特别保护区海域进行了4个航次的水质环境调查,并通过单项水质评价法、海水营养指数评价法、有机污染综合指数法等对该特别保护区海域的水质进行质量分析。调查数据表明,该保护区海水受无机氮污染严重,4个航次无机氮污染指数均大于1;海水营养指数分布范围为1．60～93．76,平均值为14．58,处于重度富营养状态;有机污染指数分布范围为0．96～6．58,平均值为2．76,处于3级轻度污染状态。结果表明,无机氮为该海域主要污染因子,富营养化及有机污染情况严重并呈增加趋势。目前保护区已经建立,但对于保护区的治理有待进一步开展,还需对该保护区进行长期的监测与科学研究。     

莱州湾蛏类生态国家级海洋特别保护区环境质量分析与评价

2017年3、5、8、10月分别对莱州湾蛏类生态国家级海洋特别保护区海水质量、沉积物质量、生物生态开展调查监测,并通过多种评价方法进行了评价。结果表明,保护区海水质量整体符合二类标准,主要污染因子为无机氮;沉积物质量总体良好,符合一类标准;保护区浮游植物生物多样性指数较低,个别站位优势种明显,存在发生赤潮可能。     

基于多个线粒体序列的中韩俄沿海不同地理群体刺参的遗传多样性及种群结构分析

为评价不同海域不同体色特征刺参群体的遗传结构,本研究采用PCR技术扩增了中国、韩国和俄罗斯沿海8个刺参(Apostichopus japonicus)群体的16S rDNA、COⅠ和D-loop序列。根据所获得的16S rDNA、COⅠ和D-loop序列分析这8个群体的遗传多样性和遗传进化关系。结果显示,16S rDNA、COⅠ和D-loop序列长度分别为543 bp、656 bp和509~527 bp。16S rDNA序列中共检测到16个多态位点,16种单倍型,单倍型多样性指数为0.629,核苷酸多样性指数为0.0016,平均核苷酸差异数0.880。COⅠ序列共检测到62个多态位点,38种单倍型,单倍型多样性指数为0.958,核苷酸多样性指数为0.0073,平均核苷酸差异数为4.796。D-loop序列共检测到200个多态位点,61种单倍型,单倍型多样性指数为0.922,核苷酸多样性指数为0.0157,平均核苷酸差异数为6.834。3个线粒体片段对于不同群体的遗传多样性检测结果显示,D-loop和COⅠ序列的多态位点数、单倍型数和核苷酸多样性均显著高于16S rDNA序列,更适用于同一物种不同群体遗传结构的解析。韩国浦项地区3个群体遗传多样性最高,这可能与其所处地理位置洋流影响有关。利用COⅠ基因对采自浦项的3种体色刺参进行遗传分化分析,遗传分化系数Fst<0.05,不存在遗传分化。对所采集的群体构建的系统进化树结果显示,青岛海参群体与烟台海参群体聚为一支,再与韩国群山黑参群体聚为一支,然后与韩国木浦黑参群体聚在一起,向外依次为俄罗斯群体及韩国浦项的3个群体,不同种群的遗传结构受洋流影响最大,其次跟其地理分布有关。     

双线紫蛤COI基因序列多态性与水环境分析

以双线紫蛤(Soletellina diphos)为研究对象,测定南日岛海区自然群体30枚个体的线粒体COI基因部分序列,研究其个体间的序列单核苷酸多态性(SNP),掌握双线紫蛤的遗传多样性;统计南日岛海域2008年4月—2018年12月期间海水样品的盐度、温度、pH、溶解氧、叶绿素a、透明度、化学需氧量、无机氮、无机磷、石油类10项理化因子检测结果,分析数据随季节月份变化状况,了解双线紫蛤栖息地水环境。结果显示,南日岛双线紫蛤群体30枚个体的序列中碱基T、C、A、G平均含量分别为41.95%、16.07%、18.53%、23.45%,(A+T)含量明显高于(G+C)含量,共发现13个核苷酸多态位点(转换/颠换偏倚率13.04),具有9个单倍型,有1个为引起亮氨酸/苯丙氨酸变化的非同义突变位点,单倍型多样性指数(Hd)为(0.598 0±0.103 0),核苷酸多样性(Pi)为(0.002 9±0.000 6),平均核苷酸变异数(k)为1.912 6,表现出略高于标准值的单倍型多样性和较低的核苷酸多样性,个体间的遗传距离为0～0.009 2、平均值为(0.002 9±0.001 0),Tajima's D和Fu's Fs中性检验的结果均为不显著负值,说明这个群体的双线紫蛤遗传相对稳定。南日岛产区水环境月均盐度变化范围为26.84～34.33、温度变化范围为13.27～29.33℃、pH变化范围为7.89～8.54、溶解氧变化范围为5.64～9.31 mg/L、叶绿素a变化范围为0.33～15.57μg/L、透明度变化范围为0.40～1.79 m、化学需氧量变化范围为0.22～1.59 mg/L、无机氮变化范围为0.04～0.45 mg/L、无机磷变化范围为0～0.16 mg/L、石油类变化范围为0～0.20 mg/L,盐度、温度在各年春、夏、秋三季变化皆先升后降,夏季最高;pH、溶解氧则在春、夏两季波动呈下降;而无机氮在春、夏、秋季趋势为先降后升,夏季最低;除了化学需氧量和石油类以外,其余参数的周年变动均表现出一定的季节性差异规律,同时表明双线紫蛤在以上的理化条件下都可以进行生长繁殖。     

双岛湾海域人工鱼礁区水质状况及其季节变化特征

根据2011年夏季(8月)、秋季(11月)和2012年冬季(3月)、春季(5月)水环境调查资料,采用单因子污染指数法、营养指数法、营养状态指数法和有机污染综合指数法对威海双岛湾海域人工鱼礁区域的水质状况进行了评价.结果表明:(1)鱼礁区溶解氧、pH、无机氮、活性磷酸盐和COD污染指数均＜1.0,海水未受到上述因子的污染;(2)鱼礁区海水处于贫营养水平,水质指标季节变化不明显;(3)鱼礁区与鱼礁影响区、对照区比较,海水营养指数、营养水平和有机污染指数无显著差异,显示鱼礁短期内对水质改善作用不明显.     

基于细胞色素b基因序列分析的华南居氏银鱼遗传多样性研究

测定了广东虎门、广西合浦和广西钦州3个群体45 ind居氏银鱼(Salanx cuvieri)线粒体DNA细胞色素b基因1 017 bp序列片段,发现42个多态位点,28个单倍型。平均单倍型多样性(Hd)和核苷酸多样性(Pi)分别为0.967和0.006,呈现出高单倍型多样性低核苷酸多样性的特点。合浦和钦州群体间遗传分化指数Fst值为-0.029,表明广西的2个群体间没有遗传分化。虎门群体与合浦和钦州群体间的Fst值分别为0.201和0.218,出现中度遗传分化,推测琼州海峡可能阻碍了广东与广西群体间的遗传交流。邻接树上出现2个无明显地理聚群的较弱谱系分支,表明正处于谱系拣选状态。AMOVA分析表明,居氏银鱼群体内遗传差异(86.54%)大于群体间(13.46%)遗传差异,遗传变异主要集中在群体内部。虎门群体中性检测的Tajima’sD为显著负值(-1.847,P=0.018),不对称分布分析呈单峰模式,表明可能在晚更新世(3.2～8.0万年前)经历过种群的快速扩张。建议广东群体和广西群体各作为一个管理保护单位分别加以保护。     

利用mtDNA COI基因序列分析引进的澳洲虫纹鳕鲈群体遗传多样性

根据线粒体COI基因序列分析了虫纹鳕鲈(Maccullochella peelii)引进群体的遗传多样性。用PCR技术扩增了线粒体COI的序列,PCR产物经纯化、克隆和测序后得到了652bp的核苷酸片段。运用CLUSTAL X(1.83)软件比对了25个个体的序列,共检测到5个多态位点,其中4个为转换位点,1个为颠换位点;运用MEGA5.0软件构建了NJ系统树;用DNASP软件计算出的单倍型个数(H)为5,单倍型多样性(Hd)为0.300,核苷酸多样性(Pi)和平均核苷酸差异数(k)为分别为0.00073和0.473。结果表明,引进群体的虫纹鳕鲈mtDNA COI基因序列的变异程度较小,群体内遗传多样性较低。     

靖海湾松江鲈鱼种质资源保护区生态环境质量分析与评价

根据2014~2016年夏季对靖海湾松江鲈鱼国家级水产种质资源保护区表层海水中营养盐和重金属等相关环境因子的连续监测数据,运用综合质量指数法对该海域生态环境质量进行综合评价,并分别采用有机污染指数法、潜在性富营养化评价法和主成分分析法从不同方面分析该海域的水质情况。结果显示,靖海湾保护区海水环境综合质量指数范围为1.04~1.69,评价等级为轻污染。有机污染指数范围为4.66~9.23,处于严重有机污染状态。根据营养化评价模式分析得出,2014年靖海湾保护区处于富营养状态,2015年和2016年均处于氮限制潜在性富营养状态。主成分分析显示,溶解无机氮(DIN)、As、Zn具有较高正载荷,为影响该海域水质的主要环境因子,应予以重点监测和防治。     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

The optimal dietary protein level of juvenile silver sillago Sillago sihama at three dietary lipid levels

A 5 × 3 factorial growth trial was conducted to evaluate optimal dietary protein and lipid levels (dietary protein level, DP; dietary lipid level, DL) for juvenile Sillago sihama (S. sihama) (2.0 ± 0.02 g, initial weight). Fish were fed 15 diets containing 5 DPs (350, 400, 450, 500 and 550 g/kg) and 3 DLs (60, 90 and 120 g/kg) for 8 weeks. The interaction between proteins and lipids significantly influenced the feed conversion ratio, condition factor, body composition, antioxidant indices and lipase activity (p < .05). DP 450 g/kg showed the highest average final body weight. DPs 500 and 550 g/kg significantly decreased the protein efficiency ratio (p < .05). DL 120 g/kg showed the highest percentage weight gain. The low feed conversion ratio was found in diets P45L12, P55L9 and P55L12. Diet P45L12 showed high superoxide dismutase activities. DP 450 g/kg showed the lowest average malondialdehyde content. Lipase activity was increased by increasing DP (p < .05) with a fall at DP 550 g/kg. Under the present experimental conditions, the optimal DP for S. sihama was 450 g/kg under the DL 120 g/kg.     

Vitamin D3 and the renal handling of phosphate in American eels

Cholecalciferol (Vitamin D₃) increased plasma inorganic phosphate concentration in American eels,Anguilla rostrata, in a dose-dependent fashion. This response was more marked in phosphate loaded fish. In control as well as phosphate loaded eels the hyperphosphatemic response to D₃ was associated with a sharp reduction in renal phosphate clearance relative to¹⁴C-polyethelene glycol (PEG) clearance. Glomerular filtration and urine flow rates were not affected by D₃. As renal phosphate clearance, even in phosphate loaded eels, never significantly exceeded that of PEG, it is suggested that D₃ reduced the relative clearance rate of phosphate by increasing renal phosphate reabsorption rather than by reducing the tubular secretion of phosphate.