Transport and accumulation of organochlorines in the ovaries of Atlantic croaker (Micropogonias undulatus)

The role of vitellogenin in the transport of organochlorines was investigated in Atlantic croaker (Micropogonias undulatus) by exposing them through the diet to o,p′-DDT at a concentration of 10.8 μg/100 g fish/day for 14 days or Aroclor 1254 (PCB) at a concentration of 0.5 mg/100 g fish/day for 30 days during gonadal recrudescence. Tissue samples were taken from the fish at various times after initial exposure, and o,p′-DDT and PCBs were extracted in acetonitrile and analyzed by gas chromatography. Analysis of the ovarian tissue collected 2 and 3 weeks from the start of exposure revealed that the o,p′-DDT concentration increases as the gonadosomatic index (GSI) increases (r² = 0.63), with accumulation ranging from less than 1% to as much as 8% of the total dosage. Interestingly, o,p′-DDT did not accumulate in the testes during the same exposure period. Accumulation of PCBs was found to be 40 times higher in the ovaries than in the testes. Gel filtration of plasma from exposed females showed that o,p′-DDT elutes in the low density lipoprotein and vitellogenin fractions. Control plasma incubated with o,p′-DDT at 4 °C for 16 h followed by chromatography on Sepharose 6B gave similar results with an o,p′-DDT concentration of 0.6μg/mg protein in the vitellogenin fraction. Furthermore, both o,p′-DDT and PCBs were found to bind to purified croaker vitellogenin. These results suggest that lipoproteins, including vitellogenin, are involved in the transport and accumulation of organochlorines in the ovaries of exposed fish.     

Reproductive endocrine function in female atlantic croaker exposed to pollutants

To examine whether xenobiotics impair teleost reproduction by altering reproductive endocrine function, steroid hormone secretion and ovarian growth were investigated in female Atlantic croaker (Micropogonias undulatus) after chronic oral administration with sublethal doses of three classes of reproductive toxins (lead, benzo[a]pyrene and Aroclor 1254). All the sublethal treatments significantly impaired ovarian growth as assessed by the gonadosomatic index. Reduced ovarian growth was accompanied by a significant decline in circulating 17β-estradiol levels in fish exposed to lead and benzo[a]pyrene. All three toxicants significantly decreased plasma testosterone levels. However, the steroidogenic capacity of ovarian tissue in vitro was not altered by xenobiotic exposure. The data suggest that the decreased ovarian growth in croaker after pollutant exposure may be a consequence of the decline in plasma 17β-estradiol levels. Further, this decrease in circulating levels of gonadal steroids does not appear to be caused by a direct effect of the chemicals on ovarian steroid synthesis.     

Disruption of neuroendocrine function in atlantic croaker exposed to Aroclor 1254

To investigate the effects of a polychlorinated biphenyl (PCB) mixture (Aroclor 1254) on reproductive neuroendocrine function, male Atlantic croaker (Micropogonias undulatus) were exposed to the PCB mixture in the diet (0.1 mg/100. g body wt/day) for 30 days during gonadal recrudescence. Concentrations of the monoamines (5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE)) and their major metabolites (5-hydroxyindolacetic acid, dihydroxyphenylacetic acid and 3-methoxytyramine) were measured in the preoptic-anterior hypothalamus (POAH) and medial and posterior hypothalamus (MPH) using HPLC with electrochemical detection. There was a significant decline in 5-HT and DA concentrations and an increase in their metabolite to parent amine ratios in both the POAH and MPH of Aroclor 1254-exposed fish. In addition, Aroclor 1254 exposure resulted in the loss of the in vitro gonadotropin (GtH) response to stimulation by a luteinizing hormone-releasing hormone analog (LHRHa). The reduced availability of 5-HT in the nerve terminals may be responsible for the loss of pituitary GtH response to stimulation by LHRHa. These results support the hypothesis that Aroclor 1254-induced alterations in pituitary gonadotropin release may be mediated, at least partially, by altered hypothalamic serotonergic activity.     

Distribution of whitemouth croaker (Micropogonias furnieri, Desmarest 1823) larvae in the Río de la Plata estuarine front

Whitemouth croaker (Micropogonias furnieri) larvae obtained and hydrographic data collected in the Rio de la Plata estuary (35°S–56°W) between 1987 and 2000 were used to explore the early life stages spatial and temporal distribution patterns and their relation to oceanographic features. The spatial distribution, restricted to a band in the inner part of the estuary, coincided with the bottom salinity front and the maximum turbidity zone (MTZ, turbidity front). Larvae were present during the warmest months (October through May) within a range of 14–24.5 °C temperature and 0.9–33 salinity. A vertically stratified sampling performed in the region where the largest abundance was found (December 2005 and March 2006) was used to test the hypothesis that larvae retention occurs in the bottom salinity front.The vertically stratified sampling showed larvae throughout the water column with high predominance in the river–estuary transition zone. A positive correlation between abundance and the bottom salinity horizontal gradient was found. The size analysis showed that the largest individuals (>10 mm SL), probably undergoing the settlement process, inhabited near the bottom and that the smallest (<10 mm SL) were present in the whole water column. Length distribution along the front showed no trend.Results support the estuarine retention hypothesis of previous studies on whitemouth croaker gravid females, eggs distribution and outcomes from a numerical simulation model. Retention in the salinity front/MTZ would allow larvae to benefit from food accumulation in the region, the high turbidity level provide shelter against predators and retention in the estuary secure closeness to the main nursery ground.     

大黄鱼EBI3基因的克隆与表达分析

通过克隆大黄鱼(Larimichthys crocea)的EBI3(Epstein-Barr virus-induced gene 3)基因并对其进行了序列分析.大黄鱼EBI3(Lyc EBI3)基因开放阅读框长747bp,编码248个氨基酸,存在1个信号肽序列和2个FN3结构域(37~130,145~230).多序列比对发现Lyc EBI3分子与其他已知EBI3氨基酸水平一致性为27.68%~57.53%.Real-time PCR结果表明,Lyc EBI3基因在脾脏和血液中表达量最高,且溶藻弧菌(Vibrio alginolyticus)刺激后,大黄鱼头肾和脾脏中Lyc EBI3基因的转录水平会显著升高,说明Lyc EBI3基因可能参与了抑制由细菌引起的炎症反应.     

Reduced survivorship of Himasthla (Trematoda, Digenea)-infected cockles (Cerastoderma edule) exposed to oxygen depletion

Bivalve populations from inshore waters often accommodate a diverse trematode fauna that may have a variety of effects on host specimens. In particular, larval trematodes that grow or reproduce within their host are known to be severe pathogens, whereas trematodes utilising bivalves only for encystment are thought to be relatively benign. Yet this may depend on the environmental conditions, and it can be expected that such trematodes in concert with other stress agents can be detrimental to host organisms. To examine the impact of such larval trematodes on hosts subjected to stress, we studied the digenetic trematode Himasthla elongata and one of its second intermediate hosts, the bivalve Cerastoderma edule. Experimentally infected cockles and non-infected cockles were exposed to oxygen depletion, whereupon we measured their burrowing ability and survivorship. After 30 h of hypoxia, the survival of infected cockles was significantly reduced compared to non-infected cockles, whereas no effect of parasites on cockles under normoxic conditions was found. In addition, parasites tended to reduce the burrowing ability of cockles exposed to hypoxia but the effect was not clear. The effect of parasites and possible ecological consequences are discussed and it is suggested that the combined effects of parasites and oxygen deficiency may explain some hitherto unexplained cases of mass mortalities in bivalve populations.     

大黄鱼( Larimichthys crocea ) CRFB13 基因的克隆与表达分析

IFN-γ是哺乳动物的一个关键免疫调节因子,通过特异性地结合IFN-γ受体1(IFNGR1)和IFN-γ受体2(IFNGR2)组成的异源四聚受体复合物,来发挥其生物学功能.目前,在鱼类中已经克隆出IFNGR1基因的2个亚型:细胞因子受体家族B 13(CRFB13)和细胞因子受体家族B17(CRFB17).本研究从大黄鱼(Larimichthys crocea)中克隆得到了一个CRFB13基因(Lyc CRFB13),其开放阅读框全长1 158个核苷酸,编码由385个氨基酸组成的1个蛋白.Lyc CRFB13蛋白具有典型的II型细胞因子受体特征,包括1个信号肽、1个胞外的FN III-like结构域,1个单次跨膜结构域,以及胞内的JAK1和STAT1结合位点.氨基酸序列分析表明鱼类的CRFB13都具有JAK1和STAT1结合位点.系统进化分析表明鱼类的2种IFNGR1,CRFB13和CRFB17形成2个独立的分支,Lyc CRFB13和鱼类的CRFB13聚在一起,与雀鲷(Stegastes partitus)CRFB13的亲缘关系最近.组织分布显示Lyc CRFB13为组成型表达,在大黄鱼鳃中的表达量最高,而在小肠中的表达量最低.Poly(I:C)刺激后,大黄鱼鳃和头肾中CRFB13的mRNA转录水平都显著升高.此外,poly(I:C)和LPS还能够诱导大黄鱼头肾白细胞中Lyc CRFB13的表达,这些结果表明Lyc CRFB13可能在大黄鱼抗病毒、抗细菌免疫反应中发挥重要作用.     

波纹唇鱼微卫星分子标记的筛选及适用性分析

采用磁珠富集法及利用生物素标记的(CA)15寡核苷酸探针从波纹唇鱼(Cheilinus undulatus)基因组DNA Bsp143Ⅰ酶切位点的400~1000 bp片段中筛选CA/GT微卫星位点。洗脱的杂交片段克隆到PMD18-T载体上构建富集微卫星基因组文库后,通过PCR筛选出阳性克隆进行测序。在120条序列中有88条序列包含有重复次数不少于5次的微卫星位点,阳性序列比例达到73.33%。其中完美型(perfect)类型微卫星最多的重复次数为26次。在88条非冗长序列中,共有28条(31.82%)微卫星重复序列两端有侧翼序列能够进行引物设计。用波纹唇鱼3个个体的混合基因进行引物筛选,其中的24对具有清晰的扩增条带。将筛选的24对引物对波纹唇鱼1个群体的39个个体进行遗传多样性分析,其中4对引物扩增产物为单态,20对扩增产物呈多态性;20对扩增多态的引物在39个个体中扩增出等位基因数为2~12,平均有效等位基因数为3.5。该波纹唇鱼群体的PIC、Ho、He的平均值分别为0.0782、0.8513、0.5667。这20个微卫星位点适于波纹唇鱼群体遗传结构的研究分析。     

Toxicity of sea-surface microlayer: Effects of hexane extract on baltic herring (Clupea harengus) and atlantic cod (Gadus morhua) embryos

This study was designed to determine whether contaminated sea-surface microlayer was toxic to marine fish embryos in its ntaive form and as a hexane extract. Developing embryos of Atlantic cod (Gadus morhua) and Baltic herring (Clupea harengus) were exposed to hexane extracts of sea-surface microlayer collected from five locations in the North Sea and Baltic Sea. Extracts from two of these locations produced significant embryos mortality as well as severe deformities in live hatched larvae. A control sample of bulk water collected from 20 cm under the surface and extracted in the same way produced no significant mortality or deformities. Significaant changes in timing of hatching were also observed in those samples which produced embryo toxic effects. A comparison of these data with those obtained from code embryos exposed to unextracted microlayer showed a similar biological effect with both unextracted samples and hexane extracts.Chemical analyses revealed the greatest biological effect in samples with petroleum hydrocarbon concentrations between 180 and > 200 μg liter⁻¹. The bulk water control had 1 μg liter⁻¹ while the three samples that showed no biological activity had 3 to 8 μg liter⁻¹ petroleum hydrocarbons. Phthalic acid esters were detected in four samples and chlorinated hydrocarbons in one, but could not be positively correlated with any of the toxic responses. No other chemical contaminants were detected in the five samples.The data presented here show that some sites contain sea-surface microlayer which can be toxic to marine fish embryos: that Baltic herring and Atlantic cod embryos respond similarly to the toxic effects of contaminated microlayer: and that unextracted microlayer and hexane extract of microlayer produce essentially the same toxic effect(s) if only organic contaminants are considered.     

大黄鱼精液的高效超低温保存

采用2 mL的冷存管和程序降温仪高效超低温保存大黄鱼(Pseudosciaena croce)精液。分析比较了5种不同浓度抗冻剂(二甲基亚砜、乙二醇、丙二醇和甘油和甲醇)、3种降温速率(10,20和30℃/min)和2种解冻温度(28℃和43℃)对大黄鱼精液超低温保存效果的影响。实验结果表明,采用Hanks’作为稀释液,10%～20%甘油或10%DMSO作为抗冻剂,20℃/min的降温速率对精液进行超低温保存43℃水浴解冻,冻精激活后获得理想的运动率(76.6%±11.4%～80.0%±12.6%),而且对10%甘油保存的冻精进行人工授精实验获得了与新鲜精液相似的受精率和孵化率。     

牙鲆对海水中铅积累排放规律研究

研究了海水中铅在牙鲆内脏、肌肉、鳃组织中吸收、积累和排放规律及海水中总有机碳浓度对铅吸收的影响.结果表明,铅浓度为0.5mg/dm3时,各组织中铅蓄积量随暴露时间增加而增大,第9天均达到吸收平衡,此时铅蓄积量(mg/kg)内脏中的649.8大于鳃中的237.4大于肌肉中的59.71.染毒13d后将牙鲆移入清洁海水中排放的结果表明,随排放时间增加各组织中铅蓄积量明显下降,第10天时各组织中铅排出率为内脏团的84.85%大于鳃的61.26%大于肌肉的57.03%.海水中总有机碳浓度对铅蓄积量有明显影响,随着总有机碳浓度升高,各组织中铅蓄积量明显下降,表明海水总有机碳能降低铅的生物有效性.     

渤海小黄鱼生殖力及其变化

根据2004年5月在渤海进行的拖网调查资料,分析了小黄鱼(Pseudosciaenapolyatics)在经过近半世纪的捕捞后的生殖群体生殖力及其与体长、纯体质量的关系,并对小黄鱼生殖力的变化作了比较。结果表明,小黄鱼的个体绝对生殖力为14729粒±1360粒;个体绝对生殖力与叉长、纯体质量的关系变化较大;与1964年历史资料相比,相同体长组小黄鱼的个体绝对生殖力F和相对生殖力FL都显著增大(P<0.01)。小黄鱼的生殖力增加可能是小黄鱼在长期捕捞等外部环境影响下的遗传进化以及对其生存环境的适应性响应。     

中国近海大黄鱼和日本带鱼群体数量变动及其资源保护措施探讨

作者根据长期调查研究,分析了大黄鱼(Larimichthys crocea)和日本带鱼(Trichiurus japonicus)群体数量变动的因果关系,并提出了加强我国海洋渔业资源科学监管的建议:(1)正确划分大黄鱼和带鱼地理种群及其产卵群体,为资源繁殖保护提供参考依据;(2)修订若干《渔业法》条款,重新审定《官井洋大黄鱼繁殖保护区管理规定》;(3)充分合理开发利用闽东渔场近海岛屿周边海域,并且采用抗风浪升降式圆柱形深水养殖网箱养殖大黄鱼;建立内湾性官井洋大黄鱼海洋牧场渔业基地,并新建中国著名舟山渔场大黄鱼繁殖保护区;(4)继续加强大黄鱼原种场建设,大力开展大黄鱼良种选育研究,每年提供大批量野生大黄鱼人工培育的原种子一代幼鱼进行增殖放流;(5)巩固和扩大伏季休渔效果,全年禁止违法而严重损害带鱼幼鱼资源的各种张网作业;(6)以法治渔,加快制定符合于我国实情的海洋渔业科学监管的有效措施.     

Stress protein expression in Ampelisca abdita (Amphipoda) exposed to sediments from San Francisco Bay

Stress proteins (heat shock proteins, hsps) form part of the cellular protein repair system, and are induced by a wide variety of Stressors. To determine their suitability as tools for assessing sublethal sediment toxicity, we measured levels of members of the stress protein families hsp60 and hsp70 in benthic estuarine amphipods (Ampelisca abdita) exposed to sediments from 23 different sampling sites in San Francisco Bay for 10 d. Concentrations of sediment-associated xenobiotics were determined. Per cent survival was recorded and surviving animals were analysed for stress proteins using western blotting techniques. An inverse correlation (r² = 0.44) was seen between amphipod survival and hsp64 levels, and hsp64 levels were positively correlated with concentrations of total polycyclic aromatic hydrocarbons (PAHs) (r² = 0.5). Principal component analysis revealed that amphipod mortality was linked to a combination of several PAHs (phenanthrene, fluoranthene, pyrene, benzo(a)pyrene) and di-n-butylphthalate at southern San Francisco Bay sites. At northern San Francisco Bay sites, negative correlations were found between hsp64 levels and organotin compounds (MBT, DBT, TBT), and between hsp71 levels and the PAHs, benzo (b,k)fluoranthene and benzo(G,H,I)perylene, suggesting an inhibitory effect of these compounds on stress protein expression.     

Biomarker responses in Macoma nasuta (Bivalvia) exposed to sediments from northern San Francisco Bay

Our study investigates biomarker responses and survival of Macoma nasuta exposed to sediments collected from six locations in northern San Francisco Bay. Biomarkers analyzed were stress proteins (hsp70) in gill, mantle and digestive gland, lysosomal membrane damage and histopathologic lesions. Sediments and clam tissues were analyzed for a comprehensive suite of heavy metals and trace organic pollutants. Sediment grain size and organic carbon content were determined. Clams accumulated metals, polyaromatic hydrocarbons (PAHs) and organochlorine pesticides (aldrin and p,p^′-DDT and its metabolites p,p^′-DDD and p,p^′-DDE). Pearson and Spearman correlation analysis revealed that mortality, hsp70 in gill and histopathologic lesion scores in gonads, and lysosomal membrane damage were significantly correlated with tissue concentrations of DDT and/or its metabolites. Tissue concentrations of metals, in particular nickel, chromium, and copper, were associated with macrophage aggregates in digestive gland and germ cell necrosis. Cadmium was linked to mortality and lysosomal membrane damage.     

Comparative studies on some genetic characteristics among four large yellow croaker (Pseudosciaena crocea) populations

The genetic structure was compared among four large yellow croaker (Pseudosciaena crocea) populations, an important aquaculture species in China. RAPD and ISSR results show that the genetic distances between domestic stocks and their wild counterparts are significantly larger than those between the three wild stocks. The sequence of mtDNA cyt b shows that the three wild populations share the same sequence, with differences in the four bases from the cultured stock: two transitions [57(C-T), 291(G-A)] and two transversions [66(C-G),223(A-C)]. Cluster analysis reveals that the domestic stock has diverged from its parental wild stock [Min—Yuedong (Fujian Province—eastern Guangdong Province, China) stock]. Results demonstrate that 20 a of domestication of the Pseudosciaena crocea stock has resulted in significant genetic changes.     

大黄鱼选育群体与普通养殖群体部分免疫指标的比较

以本课题组选育的“闽优1 号”及未经选育的普通大黄鱼(Larimichthys crocea)养殖群体为研究对象, 比较了两者的白细胞吞噬活性、血清溶菌酶活性、杀菌活力、白蛋白及免疫球蛋白含量、血清及血细胞超氧化物歧化酶(SOD)活性等免疫指标的差异。结果表明: 选育的“闽优1 号”大黄鱼血清溶菌酶和血细胞SOD 活力显著高于对照组(P<0.05), 而血清中的免疫球蛋白含量低于普通养殖群体。在病原菌攻毒后8 d 内, 选育群体累积死亡率显著低于非选育群体, 提示非特异性免疫在大黄鱼抗病免疫的早期阶段可能起着重要作用。     

大黄鱼脊椎骨及其早期发育研究

为完善大黄鱼(Larimichthys crocea)的骨骼生物学特性和以脊椎骨为数量性状的新品种选育提供理论依据,作者统计了大黄鱼成鱼脊椎骨骨节数量性状,并采用软骨—硬骨双染色方法对其成体及5～40日龄仔稚幼鱼脊椎骨骨节的发育过程进行观察,并对骨节数量和生长进行统计分析.结果表明:统计成体48尾中,骨节数主要是26,...