cDNA cloning of an aryl hydrocarbon receptor from Baikal seals (Phoca sibirica)

Species differences in sensitivity to related planar halogenated aromatic hydrocarbons (PHAH) add significant uncertainty in assessing the ecological risk to aquatic mammals. To investigate mechanisms of PHAH sensitivity in aquatic mammals, we cloned and sequenced the cDNA of Baikal seal aryl hydrocarbon receptor (AHR), an intracellular protein that initiates PHAH-mediated effects. The Baikal seal AHR cDNA has an open reading frame of 843 amino acid residues with a predicted molecular mass of 94.6 kDa. Comparison of AHR amino acid sequences indicated a high degree of sequence conservation (98%) between Baikal and harbor seals. The high conservation of AHRs between Baikal and harbor seals indicates that these seals express AHR proteins closely related structurally. In our previous report (Kim & Hahn, 2002), the dioxin-binding affinity of the harbor seal AHR was at least as high as that of the AHR from a dioxin-sensitive strain of mice, suggesting that this seal species may be sensitive to PHAH effects. This implies that Baikal seal may also be sensitive to dioxin effects.     

A comparison of the levels of hepatic aryl hydrocarbon hydroxylase in fish caught close to and distant from north sea oil fields

Large-scale use of oil-based muds for drilling operations offshore can lead to high concentrations of aromatic hydrocarbons in the sediments close to these platforms. Fish were trawled from stations close to and distant from such platforms and the levels of hepatic aryl hydrocarbon hydroxylase (AHH) were determined in cod, haddock and whiting. These three species were caught in sufficient numbers to make reasonable preliminary data sets. The data for cod and haddock showed significantly higher levels of AHH in the livers of fish caught close to oil platforms than in those caught in areas away from oil activity, while whiting showed no such differences. A much larger sample number together with some measurement of enhanced cytochrome P-450 levels would be required to substantiate this trend, but these data are the first indications that the oil in the sediments around platforms may be biochemically available to fish in the area.     

Estrogenic effect of dioxin-like aryl hydrocarbon receptor (AhR) agonist (PCB congener 126) in salmon hepatocytes

Previous studies have shown that aryl hydrocarbon receptor (AhR) agonists possess anti-estrogenic activities and several mechanisms have been proposed to explain the interactions between AhR and estrogen receptor (ER) signalling pathways. In the present study, we show that 3,3'4,4'5-pentachlorobiphenyl (PCB126 - a dioxin-like AhR agonist) produced estrogenic responses in the absence of ER agonist, in fish in vitro system. We exposed salmon primary hepatocytes to PCB126 (1, 10 and 50 pM) and the ER agonist nonylphenol (NP; 5 and 10 microM) singly and also in combination. Vitellogenin (Vtg) and zona radiata proteins (Zr-proteins) levels were analysed by semi-quantitative ELISA. We observed that the protein levels of Vtg and Zr-proteins were significantly induced in a concentration-specific manner in cells treated with PCB126 and NP, singly or in combination. In general, these results show a novel aspect of dioxin-like PCB effect not previously demonstrated in fish system.     

Identification of aryl hydrocarbon receptor 2 in aquatic birds; cDNA cloning of AHR1 and AHR2 and characteristics of their amino acid sequences

The toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and its related planar halogenated aromatic hydrocarbons (PHAHs) are mediated by the aryl hydrocarbon receptor (AHR). To investigate the potential sensitivity to PHAHs and the evolutional diversity of AHR in aquatic birds, AHR cDNAs were initially cloned and sequenced from the livers of a black-footed albatross (Diomedea nigripes) and a common cormorant (Phalacrocorax carbo). In this study, we report the identification of two distinct AHR paralog genes in these species. The two full-length AHR cDNAs from albatross were highly divergent (33% overall amino acid identity, and 60% identity in the N-terminal half). Phylogenetic analysis showed that one of them belongs to the AHR1 clade and the other one to the AHR2 clade, which has been identified only from fishes, but not yet from mammals and birds. Albatross AHR1 encoded a 861-residue protein with a predicted molecular mass of 96.7 kDa, and in the case of albatross AHR2, 925 amino acids and 100.7 kDa. From cormorant liver, the full-length AHR1 cDNA and the partial AHR2 cDNA were cloned. This result strongly suggests that bird species also possess two distinct AHR genes (AHR1 and AHR2). To our knowledge, this is the first report on the presence of an AHR2-like isoform in bird species as well as in fish.     

Relative importance of hydrocarbon pollutants, salinity and tidal height in colonization of oyster reefs

The relative effects of hydrocarbon pollutants, salinity and tidal height on the invertebrates and fish that inhabit oyster reefs were studied along the Louisiana Gulf of Mexico coastline. Dried oyster shell (cultch) was first exposed to crude oil in the laboratory. In a series of experiments, plastic trays filled with control and oil-exposed cultch were then deployed at two locations differing in salinity, in two seasons and at two tidal levels. In experiments on hydrocarbon, salinity, and seasonal effects, trays were colonized for one month. To examine the effects of colonization time, half of the trays were retrieved after two and the rest after five weeks. Salinity dramatically affected oyster reef assemblages, with species richness and total abundance halved at the estuarine site. Hydrocarbon effects were less prominent, whether cultch was dosed with light or heavy crude oil. The sub-tidal site had higher colonization rates, but colonization interval did not affect colonization, and seasonal differences occurred only at the higher-diversity, sub-tidal site. To determine effects of cleaners, Corexit 9580 was applied alone and with oil on cultch, and trays were colonized for one month. At high concentrations, the cleaner ameliorated hydrocarbon effects. In general, hydrocarbon effects were less prominent than salinity and aerial exposure in explaining colonization of oyster reef assemblages. Gas chromatography/mass spectrometry analysis of oyster shells after one month immersion revealed considerable losses of oil, especially with higher flow at the inter-tidal site. Sediment on shell also diluted oil. We argue that oyster reef assemblages should recover from small-scale spills, unless they occur during periods of reproduction and dispersal.     

三都湾网箱养殖鱼类体石油烃含量水平及其分布特征

本文报道了福建三都湾网箱养殖区的3种主要养殖鱼类体石油烃含量水平.结果表明:三都湾3种网箱养殖鱼类大黄鱼、鮸鱼和鲈鱼体中石油烃含量的范围是(0.69~7.48)×10-6(m/m,湿重,下同),总平均值3.63×10-6(m/m).3种网箱养殖鱼类体石油烃含量水平为大黄鱼>鲈鱼体>鮸鱼.养殖鱼类石油烃含量明显低于养殖贝类的含量,区域性和季节性的差异对养殖鱼类石油烃含量具有一定的影响.文中还将本研究结果与国内外其它海域鱼类石油烃含量水平作了比较.     

多环芳烃化合物对栉孔扇贝组织芳烃羟化酶活力的影响

研究了苯并(a)芘(B[a]P)、苯并(k)荧蒽(B[k]F)以及两者的混合物对栉孔扇贝(Chlamys farreri)鳃丝和消化盲囊芳烃羟化酶(AHH)活力的影响。结果表明,多环芳烃化合物(PAHs)对栉孔扇贝鳃丝和消化盲囊AHH活力具有显著的诱导作用(P<0.05),并且呈现了明显的时间、剂量效应特征。B[a]P和B[k]F混合物处理组鳃丝和消化盲囊的AHH活力在实验期间呈现先升高后稳定趋势,B[a]P处理组则表现峰值变化,然后呈下降趋势。消化盲囊各处理组AHH活力显著高于鳃丝。作者认为可以采用消化盲囊的AHH活力来反映周围环境PAHs的污染,并应以最大饱和浓度为标准。3种PAHs中B[a]P的毒性最强,PAHs对栉孔扇贝的毒性大小与PAHs的组成有关,不能以不同种类单一PAHs毒性的总和来代表PAHs混合物的毒性。     

应用鱼类生物标志物和IBRv2监测与评价泉州湾石油类污染生物效应状况

以褐菖鲉(Sebastiscus marmoratus)细胞色素P4501A四种生物标志物,即鳃丝EROD活性、肝EROD活性、肝CYP1A mRNA和肝CYP1A蛋白为指标,在泉州湾海域选择6个站位(包括1个对照站位),开展了为期两周的野外监测实验,研究了上述生物标志物的变化特征以及它们同表层海水中石油类含量和沉积物物中16种优先监控的PAHs含量之间的相关性,并利用新一代综合生物标志物响应指数法(IBRv2)对监测站位石油类和PAHs污染效应进行了评价.结果显示,不同站位的上述四种生物标志物变化趋势基本一致;生物标志物的变化与相应站位表层水中石油类含量及沉积物中PAHs总含量相吻合;IBRv2分析结果表明,各监测站位IBRv2指数介于1.5~4.2之间,其中2号站环境污染压力最大,4号站最小.在复杂的海洋污染条件下,采用CYP1A多种生物标志物监测技术能够更加全面反映石油类等污染及其生物效应状况.     

Relative and combined effects of habitat and fishing on reef fish communities across a limited fishing gradient at Ningaloo

Habitat degradation and fishing are major drivers of temporal and spatial changes in fish communities. The independent effects of these drivers are well documented, but the relative importance and interaction between fishing and habitat shifts is poorly understood, particularly in complex systems such as coral reefs. To assess the combined and relative effects of fishing and habitat we examined the composition of fish communities on patch reefs across a gradient of high to low structural complexity in fished and unfished areas of the Ningaloo Marine Park, Western Australia. Biomass and species richness of fish were positively correlated with structural complexity of reefs and negatively related to macroalgal cover. Total abundance of fish was also positively related to structural complexity, however this relationship was stronger on fished reefs than those where fishing is prohibited. The interaction between habitat condition and fishing pressure is primarily due to the high abundance of small bodied planktivorous fish on fished reefs. However, the influence of management zones on the abundance and biomass of predators and target species is small, implying spatial differences in fishing pressure are low and unlikely to be driving this interaction. Our results emphasise the importance of habitat in structuring reef fish communities on coral reefs especially when gradients in fishing pressure are low. The influence of fishing effort on this relationship may however become more important as fishing pressure increases.     

The value of Pacific sardine as forage fish

This paper develops a simple bioeconomic model to investigate the economic and ecological issues associated with the commercial harvest of Pacific sardines relative to their value as forage for commercially, recreationally, and ecologically important predators in the California Current ecosystem. The model was used to evaluate how changes in the net per unit value of sardines, the net per unit value of sardine predators, and the transfer efficiency of predators affect the total net value of sardines, i.e. the net value of catches and the net value from sardine predation. Given recent market conditions for sardines and their commercial predators, and transfer efficiencies derived from predation data of the 1960s, it was found that the value of commercially caught predators and the efficiency by which they convert sardines to exploitable biomass were most important in determining the viability of the sardine fishery. The values assumed for predators that are not commercially caught were of no consequence under these conditions. Taking the value of sardines as forage into account does not necessarily mean an either-or situation for the fishery. As long as there is some measure of net value from the fishery and net value from predation, both benefit society at large.     

Formation and identification of PAHs metabolites in marine organisms

In this study, the development of the technique APCI⁺ LC/MS/MS allowed the detection of phenanthrene, pyrene and metabolites of alkyl homologs in fish bile (in situ) and in urine of crabs. Laboratory experiments were carried out exposing crabs from an unpolluted mangrove (Barra de Guaratiba) to phenanthrene, and to the alkylated homologs 1-methyl phenanthrene and 2,6,9-trimethyl phenanthrene. Urine samples were collected at 0, 24, 48, 72, and 96 h. Fishes were captured from strategic sites from Guanabara Bay. Hydroxylated metabolites of phenanthrene, epoxides, orthoquinone and glucoside conjugates were identified in both samples. The method APCI⁺ LC/MS/MS showed to be effective in a preliminary assessment of phenanthrene metabolite formation, although the low concentrations of 1-methyl phenanthene and 2,6,9-trimethyl phenanthrene did not allow a systematic evaluation of data. The method, however, proved to be excellent tool for studies of PAHs metabolites due to the high selectivity, sensitivity and separation attained.     

胶州湾海域PAHs 污染与可培养半知菌的多样性

用5种不同的培养基对胶州湾近岸污染环境可培养半知菌进行选择性分离,共得到235株半知菌,其中YM培养基、MPDA培养基对半知菌的分离效果最好,分别占分离总量的26.4%和24.7%。试验选择的3个站位：石老人潮间带、海泊河入海口、李村河入海口PAHs含量存在显著性差异(P<0.05),其中石老人最低,平均为113.8 ng/g,海泊河 PAHs 为324.9 ng/g,介于中等水平,李村河含量最高为882.1 ng/g。可培养半知菌群体随PAHs含量的增加表现出先升高后降低的趋势。对各站位的半知菌进行统计发现,污染不同的站位,可培养半知菌优势种群不同,说明PAHs的污染可对半知菌的菌落结构组成产生影响。     

Oyster patch reefs as indicators of fossil hydrocarbon seeps induced by synsedimentary faults

The Late Jurassic deposits of the Boulonnais area (N-France) represent the proximal lateral-equivalent of the Kimmeridge Clay Formation; they accumulated on a clastic-dominated ramp subject to synsedimentary faulting as a result of the Atlantic Ocean rifting. In the Gris-Nez Cape area, i.e., close to the northern border fault zone of the Jurassic basin, the Late Jurassic sequence contains small-dimensioned oyster patch reefs (<1 m) that are specifically observed at the base of an abrupt deepening trend in the depositional sequence induced by well-defined pulses of normal fault activity. Petrographic analysis of these patch reefs shows that they are exclusively composed of Nanogyra nana embedded in a microsparitic calcite matrix. ™¹³C measurements, carried out within both the matrix and the shells, display significantly lower values in the matrix compared to the oyster shells which suggests that the carbonate matrix precipitation was involving a carbon source different from marine dissolved inorganic carbon, most probably related to sulfate reduction, which is evidenced by light ™³⁴S in pyrites. Similarities but also differences with lucinid-rich bioconstructions, namely, the Late Jurassic pseudo-bioherms of Beauvoisin (SE-France) suggest that the patch reefs developed at hydrocarbon seeps are related to synsedimentary faults. The extensional block-faulting segmentation of the northern margin of the Boulonnais Basin in Late Jurassic times is thus believed to have induced a sort of small-dimension hydrocarbon seepage field, recorded by the patch reef distribution.     

Assessing and monitoring local and long-range-transported hydrocarbons as potential stressors to fish stocks

The Arctic is poorly described and understood compared to the North Sea and other boreal areas. We need to learn how knowledge obtained at boreal conditions may be applied at Arctic conditions. Increasing activity of the hydrocarbon industry may cause exposure stress from discharges. Within a joint industry research programme (Biosea) we studied how responses in fish from oil exposure may differ at North Sea boreal and Barents Sea Arctic conditions. Hydrocarbon uptake, metabolites, and enzymatic and genotoxic biomarker type of effect responses were measured in cod (Gadus morhua L.). Hydrocarbon metabolites remain longer in fish bile than original hydrocarbons, which are eliminated fast from tissues. The metabolites may be measured to background concentrations. They describe exposure and they constitute a link to other effects. Body burden, biliary polyaromatic hydrocarbon (PAH) metabolite concentration, and quantities of Cytochrome P-450 1A (CYP1A) and DNA adducts increased with oil in water concentration. The extent of biomarker expression was lower for some biomarkers and elimination was slower at the lowest temperature. The results show that several factors have to be accounted for if warm-water biomarker data are to be applied in cold water.     

Carbonyl reductase mRNA abundance and enzymatic activity as potential biomarkers of oxidative stress in marine fish

Carbonyl reductase (CBR) is an enzyme involved in protection from oxidative stress. In rainbow trout (Oncorhynchus mykiss), the hepatic mRNA abundance of the two isoforms (A and B) is increased after exposure to treated sewage effluents, as well as after exposure with β-naphthoflavone (β-NF) and the pro-oxidant paraquat. In this study, we show that the same chemicals similarly increase the single known hepatic CBR mRNA level and CBR catalytic activity in the coastal living eelpout (Zoarces viviparus). Hepatic CBR mRNA abundance and catalytic activity were also compared between eelpout collected at contaminated and reference sites on the Swedish west coast, but no differences were observed. In conclusion, CBR is a potential biomarker candidate for monitoring the exposure and effects of AhR agonists and/or pro-oxidants in the marine environment, but more research is needed to investigate temporal regulation as well as dose dependency for different chemicals. The mRNA and enzymatic assays presented in this study provide two additional tools for researchers interested in expanding their biomarker battery.     

重组鲈鱼生长激素酵母饲料添加剂及对网箱喂养海水鱼的促长效应

该文应用基因工程方法开展了具有加速鱼生长的鱼生长激素基因工程研究，构建了高效表达的66鱼生长激素甲醇半赤酵母工程菌．研究中试规模的培养基配方，解决中试工艺设计和生产试验及质量检验方面的问题；通过发酵培养该工程茵获得重组鱼生长激素产品，研究掺入饲料的合适比例，并进行生产规模的海水经济鱼类喂养试验．先后对花尾胡椒鲷(Plectorhinchus cinctus)，卵形鲳鯠(Trachinouts ovatus)，大黄鱼(Pseudosciaena croces)等3种海水经济鱼(4批共约6万尾)进行生长状况对比，增重比率均值分别为20．0％、17．3％、18．4％和56．8％．饲料添加酵母工程菌具有明显地促进海水鱼生长的作用。