Comparison of toxicity effects of fuel oil treated by different dispersants on marine medaka (Oryzias melastigma) embryo

This study aims to evaluate the subacute toxic effects of oil under different treatments on marine organism by simulating natural contaminative processes. In this study, 120# (RMD15) fuel oil was selected as the pollutant and marine medaka (Oryzias melastigma) embryos as the experimental organism. The developmental toxicity of different volume concentrations (0.05%, 0.2%, 1% and 5%) of water-accommodated fractions, biologically-enhanced water-accommodated fractions, and chemically-enhanced water-accommodated fractions on the embryos in different exposure time (8, 15 and 22 d) were compared and the content of relevant polycyclic aromatic hydrocarbons (PAHs) was studied (in dispersion and in vivo). The subacute toxic effects were assessed in terms of antioxidant activities of enzymes (superoxide dismutase, catalase and glutathione S-transferase) and the blue sac disease (BSD) indexes.The results showed that the BSD indexes of the treatment groups were significantly higher than the respective control groups and showed positive correlations with both concentration and exposure time. The experiments with three antioxidant enzymes indicated that enzymatic activities of the embryos changed dramatically under the oxidation stress of petroleum hydrocarbons, especially after adding the dispersants. With the increase of petroleum hydrocarbon concentration and exposure time, the three enzymes showed different degrees of induction and inhibition effects.     

外源激素对养殖牙鲆血浆睾酮和雌二醇含量的影响研究

雄性牙鲆单独注射促黄体激素释放激素类似物(LHRH-A)(每千克质量的注射量,下同)25μg/kg或人绒毛膜促性腺激素(hCG)1 000 IU/kg,血浆睾酮(T)含量在注射后6 h大幅度降低,12h时达到最低值,以后逐渐回升;LHRH-A与hCG联合注射[LHRH-A(12.5μg/kg)+hCG(500 IU/kg)],注射后6 h血浆T含量剧烈下降,24 h时达到最低点,以后缓慢回升;对照组雄鱼血浆T含量随着采样进行而呈现降低趋势.雌鱼单独LHRH-A后,血浆雌二醇(E₂)水平逐渐下降,24 h时达到最低值,30 h后略有回升;单独注射hCG后,血浆E₂含量缓慢下降,30 h达到最低值;LHRH-A与hCG联合注射,注射后6 h血浆E2含量逐渐下降,24 h时达到最低点,30 h时缓慢回升.对照组雌鱼血浆E₂水平也随着采样的进行而逐渐降低.结果表明:单独或联合注射LHRH-A和hCG降低了雌、雄牙鲆血浆E₂和T含量,与对照组结果相同.     

The Japanese medaka (Oryzias latipes) model: applicability for investigating the immunosuppressive effects of the aquatic pollutant benzo[a]pyrene (BaP)

Despite the fact that BaP is a carcinogen, mammalian immunosuppressant, and ubiquitous aquatic pollutant, knowledge regarding the effects of BaP on the immune system of fish is still lacking. To begin to fill this gap, studies were conducted in medaka to examine the effects and mechanisms by which BaP exposure might alter host immunocompetence. Fish, exposed by IP injection of BaP (2-600 microg/g BW), were examined after 48 h for effects upon immune function and CYP1A expression/activity. Benzo[a]pyrene, at a concentration below that which increased levels of CYPIA expression/activity (2 microg BaP/g BW) suppressed lymphocyte proliferation. Concentrations of BaP at 20 and 200 microg/g BW. suppressed antibody-forming cell (AFC) numbers, superoxide production, and host resistance against bacteria. In contrast, exposure to the low affinity aryl hydrocarbon receptor (AhR) agonist, benzo[e]pyrene (BeP), neither induced CYP1A expression nor altered immune function. Given the lack of immunosuppressive effects produced by BeP, and the fact that exposure to the AhR antagonist (and CYP1A inhibitor) alpha-naphthoflavone (ANF) ameliorated the suppressive effects of BaP upon AFC numbers, the AhR pathway (including CYP1A-mediated production of reactive BaP metabolites) appears important in mediating BaP-induced immunotoxicity in fish, as in mammals. In the past, the medaka has proven a successful model for assessing carcinogenic agents. These studies have demonstrated its utility for also determining the immunosuppressive effects of an important aquatic contaminant.     

自然环境下虾夷扇贝(Patinopecten yessoensis)体内麻痹性贝毒的净化研究

为探究在自然环境中虾夷扇贝Patinopecten yessoensis体内麻痹性贝毒(PSTs)的净化过程,从黄海北部大窑湾海域采集了受PSTs污染的虾夷扇贝,将其转移至黄海的棋盘磨水域,该水域未曾有PSTs污染的报道。本研究采用高效液相色谱-荧光检测法(HPLC-FLD)检测30天净化过程中虾夷扇贝体内PSTs的组分和水平。研究结果发现,在第9天,虾夷扇贝软体中的PSTs的含量下降到一个相对较低的水平。而且,初期扇贝体内PSTs的净化率高于后期。扇贝各组织器官中PSTs含量分析发现,消化腺中的PSTs含量最高,这对于人类健康和扇贝养殖具有重要的意义。在本研究中还发现,虾夷扇贝的死亡率与PSTs的水平有一定的关系。     

三个牙鲆育种群体亲本效应及遗传参数估计

针对牙鲆(Paralichthys olivaceus)养殖品种生长速度变慢、抗病力降低这一问题。近年来构建了牙鲆抗病群体(RS)、日本群体(JS)和黄海群体(YS)3个育种群体,开展牙鲆选择育种,建立了63个家系,收集了所有家系5 328尾鱼苗的体长和体重数据。借助线形混合模型(Linear mixedmodel)、加性-显性模型、约束极大似然法(REML)及最佳线性无偏预测(BLUP)分别估计亲本效应值、体重和体长遗传力和遗传相关、父母本效应和杂交效应。预测结果显示,其中父本130,141,146,170,218,229,303和308随机效应达到了极显著效应水平(P<0.01),父本112,122,141,190,212达到显著水平(P<0.05),所有母本体重随机效应达到了极显著水平(P<0.01);体长和体重两性状的加性、显性、表型和基因型相关分量都达到极显著水平(P<0.01),体长和体重的加性方差比率均大于显性方差比率,可见两性状的遗传效应主要取决于基因的加性效应。体长的总遗传效应率为31.20%,体重的总遗传效应率为33.58%,体长和体重的随机方差比率分别达到68.80%和66.41%,说明养殖环境等因素对牙鲆体长和体重的影响较大,同时说明在牙鲆选择育种中消除环境影响极为重要。体长和体重的狭义遗传率分别为0.199和0.256(P<0.01),广义遗传率分别为0.312和0.336(P<0.01),牙鲆在以上两性状上属于中等遗传力,通过选择育种可取得较大的遗传进展。对3个牙鲆育种群体杂交随机效应估计结果显示,抗病群体和黄海群体在体长和体重两性状上加性随机效应显著(P<0.05);抗病群体分别与日本群体、黄海群体杂交,其后代表现出显性随机效应达到0.10的水平(P<0.10),黄海群体自交、日本群体和黄海群体杂交随机效应不显著(P>0.05),因此在杂交组配中选择抗病群体与其他两个群体交配可取得较好的效果。     

Feeding ecology of Japanese Spanish mackerel(Scomberomorus niphonius) along the eastern coastal waters of China

Feeding activities provide necessary nutrition and energy to support the reproduction and development of fish populations. The feeding ecology and dietary plasticity of fish are important factors determining their recruitment and population dynamics. As a top predator, Japanese Spanish mackerel(Scomberomorus niphonius) supports one of the most valuable fisheries in China. In this study, the feeding ecology and diet composition of Japanese Spanish mackerel spawning groups were analysed based on samples collected from six spawning grounds along the eastern coastal waters of China during spring(March to May) in 2016 and 2017. Both stomach contents and stable isotope analysis were conducted. Stomach content analysis showed that spawning groups of Japanese Spanish mackerel mainly fed on fish, consuming more than 40 different prey species. Diets were significantly different among sampling locations. The most important prey species were Stolephorus in Fuzhou, Japanese anchovy Engraulis japonicus in Xiangshan, Euphausia pacifica in Lüsi, sand lance Ammodytes personatus in Qingdao and Weihai, and Leptochela gracilis in Laizhou Bay. Stable isotope analysis showed that the trophic level of Japanese Spanish mackerel was relatively high and generally increased with latitude from south to north. In the 1980 s, the diet of Japanese Spanish mackerel was dominated solely by Japanese anchovies in the eastern coastal waters of China. The results in the present study showed that the importance of Japanese anchovies declined considerably, and this fish was not the most dominant diet in most of the investigated waters. Both the spatial variations in diet composition and changes in the dominant diet over the long term indicated the high adaptability of Japanese Spanish mackerel to the environment. Combining the results of stomach analysis and stable isotope analysis from different tissues provided more comprehensive and accurate dietary information on Japanese Spanish mackerel. The study provides essential information about the feeding ecology of Japanese Spanish mackerel and will benefit the management of its populations in the future.     

Surflan and oryzalin impair reproduction in the teleost medaka (Oryzias latipes)

We conducted studies to determine if the xenoestrogens Surflan and its active ingredient oryzalin, affect indices of reproductive fitness in medaka (Oryzias latipes). Oryzalin (0.5, 0.25mg/l) or Surflan (2.0 microl/l) and oryzalin (0.5mg/l) significantly increased the mean number of non-fertilized eggs produced by treated females paired with untreated males, or by untreated females paired with treated males. Oryzalin (1.0, 0.5, 0.25mg/l) and Surflan (3.8, 2.0, 1.0 microl/l) significantly affected the time to hatch of eggs from treated females paired with untreated males, and from untreated females paired with treated males. Surflan (3.8, 2.0, 1.0 microl/l) induced intersex lesions in 80-100% of males. Oryzalin-exposed males exhibited a significant increase in the incidence of necrotic spermatids and necrotic spermatogonia, while oryzalin-exposed females had significantly fewer immature oocytes and an increase in the occurrence of hyperplastic ovaries. Our results indicate that Surflan and oryzalin affect both reproduction and gonadal histology in male and female medaka.     

Carbon, nitrogen and phosphorus stoichiometry in Japanese anchovy (Engraulis japonicus) from the Huanghai Sea, China

Generally, nutrient cycle is closely related to the element distribution in biomass and the population dynamics in ecosystems. Carbon, nitrogen and phosphorus in Japanese anchovy (Engraulis japonicus) of different body lengths from the Huanghai Sea (Yellow Sea) were determined to better understand their variability and reasons during its life history. The mean content was 45.12%, 10.12% and 2.02% for C, N and P, respectively. Significant differences in C, N and P elemental composition were found among different sizes, which could be explained by varying proportions of storage compounds in whole body fish, and varying degrees of ossification. Considering abundant resources in Japanese anchovy, it was an important P-pool in the cycle of P. Moreover, the excreted N/P ratio was significantly different in fishes of different sizes, especially at high gross growth efficiency. In the past two decades, overexploitation tended to cause smaller body length in the community structure of anchovy, which presumably changed the nutrient cycle in food webs of the Huanghai Sea. Exptrapolation of the results indicates that Japanese anchovy may be important for conveying nutrient in the Huanghai Sea.     

Effect of estradiol-17β on pituitary-thyroidal axis of Japanese eel (Anguilla japonica)

In teleost sPeCies, it is well accepted that estradiol--l75 (E2 )is generated from ovarian fOl-licles stimulated by pituitary gonadOtropins (GTH) (Nagahama, l987). FUrthermore, E2 canstimulate hepatic vitellngenesis and elevate plasrna yOlk protein precursor vite1lOgenin, totalCa and W levels in several teleosts (KorSgaard and Petersen, l976, l978; Whiteheadet al., l978; Elliott et al., l979; Pertersen et al., l983; Wallace, 1985; tapigliati etal., 1999). On the other hand, many stud…     

Response of Japanese anchovy (Engraulis japonicus) to the Pacific Decadal Oscillation in the Yellow Sea over the past 400 a

Quantitative identification of long-term changes in the abundance of Japanese anchovy (Engraulis japonicus) in the Yellow Sea is particularly important for understanding evolutionary processes of the Yellow Sea ecosystem. Unfortunately, the driving mechanisms of climate variability on the anchovy are still unclear due to the lack of long-term observational data. In this study, we used the fish scale deposition rate in the central Yellow Sea to reconstruct the time series of the anchovy stock over the past 400 a. On this basis, we further explored the impacts of the Pacific Decadal Oscillation (PDO) on the anchovy. Our results show that the anchovy stock is positively correlated with the PDO on a decadal time scale. In addition, anchovy abundance was relatively high during 1620–1860 AD (the Little Ice Age, LIA), though in a state of constant fluctuation; anchovy abundance maintained at a relatively low level after ~1860 AD. In particular, followed by overfishing since the 1980s, the anchovy stock has declined sharply. Based on these findings, we infer that fluctuations of the anchovy stock may be regulated by basin-scale “atmosphere–ocean” interactions. Nevertheless, the role of overfishing should not be ignored.     

Cloning of cytochrome P450 1A (CYP1A) genes from the hermaphrodite fish Rivulus marmoratus and the Japanese medaka Oryzias latipes

To use two small fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) and the Japanese medaka Oryzias latipes (Belloniformes) as testing models in molecular ecotoxicology, we have cloned the cytochrome P450 1A (CYP1A) gene after screening of both genomic DNA libraries, and sequenced 11,863 and 7,243 bp including all the exons and introns with promoter regions, respectively. The Rivulus and the medaka CYP1A gene consisted of seven exons (including non-coding exons) with high homology to mammals. In the promoter region, Rivulus CYP1A gene has seven xenobiotic response elements (XREs) and two metal response elements (MREs), while the Japanese medaka CYP1A gene has six XREs and four MREs. Interestingly, medaka CYP1A gene has a number of MREs at the promoter, which may affect its response on metal exposure. We describe here the gene structure of both fish CYP1A genes.     

雌牙鲆CYP19A基因启动子“CpG”岛区的单核苷酸多态性及与繁殖性能的关联分析

芳香化酶基因(CYP19)编码的芳香化酶是雄激素转化为雌激素的关键酶,在鱼类性腺发育及繁殖内分泌过程中起着重要的作用。本研究采用聚合酶链式反应-单链构象多态(PCR-SSCP)和放射性免疫方法(RAI)对61尾雌牙鲆进行了CYP19A基因启动子CpG岛区多态性检测,并将其与繁殖性能进行了关联分析。CpG岛区存在3种类型的基因型(pattern1、pattern2、pattern3),经测序发现CpG岛区有以下突变:T-1927插入或缺失、C-1899插入或缺失、T-1772C、C-1766T、TGTCAAC-1752~-1744插入或缺失(11个SNPs连锁突变)。方差分析结果表明:此位点对雌二醇(E2)水平的效应达到显著(P<0.05)水平,经多重比较分析,具有pattern1基因型的个体的雌激素(E2)水平显著低于pattern2和pattern3基因型个体的雌激素水平(P<0.05),同时pattern1基因型的个体的mRNA表达量显著低于pattern2和pattern3基因型个体的mRNA表达量(P<0.05)。以上结果表明,CYP19A基因调控区甲基化位点突变可能导致芳香化酶的表达量,进而影响牙鲆繁殖内分泌过程;同时也阐明了这些与牙鲆繁殖性状相关的SNPs可以用于生产实践中进行标记辅助选择具有优良繁殖性能的牙鲆。     

西北太平洋海域太平洋褶柔鱼秋生群产卵场指数动态评估

为探究太平洋褶柔鱼(Todarodes pacificus)秋生群资源丰度波动的原因,本研究利用适宜产卵的海表温度(SST)和水深数据,构建了太平洋褶柔鱼秋生群1979—2018年40年潜在产卵场模型,在此基础上开发了适宜海表温度均值(MVSS)、适宜性海表温度加权面积(SSWA)和等温线经向位置(MP)3种产卵场指数...     

条石鲷鳔器官早期发育的组织学观察

应用石蜡切片技术和显微测量法,对条石鲷(Oplegnathus fasciatus)鳔的早期发生、分化和形成过程进行了连续观察。结果显示:条石鲷为闭鳔鱼类,在水温为23.5℃±0.5℃的条件下,鳔的形成过程可分为鳔原基出现、鳔管形成、鳔腔充气、鳔管退化及气腺形成4个时期。1日龄(孵化后第2天)仔鱼,在食道与肠的交界处分化出鳔原基;4日龄,鳔管原基出现;5日龄,气腺原基出现;6日龄,迷网出现;8日龄,鳔管形成;10日龄,鳔开始充气;17日龄,鳔充气完毕,鳔管开始退化;20日龄左右,鳔管从幽门处完全脱离;25日龄,气腺迷网发育完全;30日龄,鳔管完全消失,鳔的发育基本完善。在条石鲷的苗种培育过程中,鳔器官的发育影响到条石鲷仔鱼的成活率。     

Changes in serum thyroid hormone levels and thyroid gland activity of artificially maturing female Japanese eel(Anguilla japonica)

In order to evaluate the involvement of the thyroid gland in reproduction, thyroid functioning was described in female cultivated Japanese eel (Anguilla japonica) in progressive stages of sexual maturation induced by chum salmon pituitary homogenate (SPH) treatment. Serum thyroid hormones, thyroxine (T4) and triiodothyronine (T3) were also measured in each stage. Thyroid gland activity (epithelial cell height) was high before SPH injection (previtellogenic stage), further increasing at the early vitellogenic stage, thereafter decreasing to late vitellogenic and migratory nucleus stages. The profiles of both T3 and T4 changed during vitellogenesis, being high during previtellogenic and early vitellogenic stages, and subsequently declining, thus mimicking thyroid gland activity. These results suggest that the thyroid has relation with eel ovarian development during artificial maturation.     

鲈鱼生长催乳素的分离纯化与N-末端序列分析

生长催乳素(SL)是生长激素/催乳素家族的一种新的脑垂体蛋白质激素,本研究首次从鲈鱼脑垂体中分离得到.鲈鱼脑垂体在碱性条件下,经葡聚糖凝胶(SephadexG-100)过滤和反相高效液相色谱(rpHPLC)分离出纯化的鲈鱼生长催乳素,(sbSL),并与大麻哈鱼生长催乳素(sSL)抗体发生特异性的Western免疫印迹交叉反应证实.SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析,发现鲈鱼生长催乳素分别是由分子量为23.2×103和28.4×103u两种形式组成(后者可能是一种糖基化形式).根据Edman降解原理,测得鲈鱼生长催乳素的N-末端16个氨基酸的序列,它与已知鱼类生长催乳素的N-末端序列比较,具有较高的同源性;由此进一步证实,分离纯化得到的鲈鱼生长催乳素是正确的.     

Gene structure of the novel cytochrome P4501D1 genes in stickleback (Gasterosteus aculeatus) and medaka (Oryzias latipes)

The cytochrome P450 1 (CYP1) family has expanded with the addition of the CYP1B and CYP1C subfamilies. We recently identified a new CYP1 subfamily in zebrafish, CYP1D, with a single gene, CYP1D1. Here we examined sequences found in other fish genomes, i.e., stickleback (Gasterosteus aculeatus) and medaka (Oryzias latipes), for similarities among fish CYP1D1 genes. The full-length deduced amino acid sequences for CYP1D1 in these two species averaged about 43% identity to the CYP1As, but nearly 50% when sequence alignment ambiguities were masked. CYP1D1 has seven exons, similar in size and position to the exons in CYP1D1 and CYP1A in zebrafish. However, the intronic distances were substantially smaller in the medaka and stickleback. There also were differing numbers of putative xenobiotic response elements in the CYP1D1 of the various species. Whether the stickleback or medaka genes are inducible by aryl hydrocarbon receptor (AHR) agonists is yet to be determined.