Spatial variation of metal bioaccumulation in the hydrothermal vent mussel Bathymodiolus azoricus

The variability of the bioaccumulation of metals (Ag, Cd, Cu, Fe, Mn and Zn) was extensively studied in the mussel Bathymodiolus azoricus from five hydrothermal vent sites inside three main vent fields of increasing depth along the Mid-Atlantic Ridge: Menez Gwen, Lucky Strike and Rainbow. Metal bioaccumulation varied greatly between vent fields and even between sites inside a vent field with B. azoricus showing a great capacity to accumulate metals. The bioaccumulation of these metals also varied significantly among tissues. The main target was the gills where metals were mainly associated with soluble compounds whereas in the digestive gland they were mainly associated with insoluble compounds. Storage of metals under insoluble forms in B. azoricus seems to be a major pathway for the detoxification of both essential and non-essential metals. Mussels from the studied fields can be discriminated following their metallic load but the segregation relies partially on the composition of the metal-enriched fluids.     

Seasonal variations in lipid composition of the hydrothermal vent mussel Bathymodiolus azoricus from the Menez Gwen vent field

Specimens of the hydrothermal mussel Bathymodiolus azoricus collected in Menez Gwen hydrothermal vent field (NE Atlantic) during 2002–2003 were examined for feeding patterns variations through three seasons. The fatty acid profile and lipid classes of the mussels were studied, together with the MODIS/AQUA-derived near-surface chlorophyll a to test the hypothesis that surface productivity might be related to the feeding patterns of this species. The lipid levels showed pronounced seasonal fluctuations with the highest values occurring in January and August. Seasonal variations in lipid classes and fatty acid composition of neutral and polar lipids in the mussels are presented. Differences in the fatty acid profile of lipid classes in different seasons suggest that the higher energy requirements in summer and winter were supplied by bacterial biomarkers ω7 MUFA (monounsaturated fatty acids), whereas ω6 PUFA (polyunsaturated fatty acids) and NMI (non-methylene-interrupted) fatty acids predominated during the spring. The MODIS/AQUA data show marked seasonal variability and an anomalous peak during January of 2003, although this cannot be directly linked to lipid composition variation.     

Temporal variation in the antioxidant defence system and lipid peroxidation in the gills and mantle of hydrothermal vent mussel Bathymodiolus azoricus

Hydrothermal vent mussels are exposed continually to toxic compounds, including high metal concentrations and other substances like dissolved sulphide, methane and natural radioactivity. Fluctuations in these parameters appear to be common because of the characteristic instability of the hydrothermal environment. Temporal variation in the antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidases (Total GPx), selenium dependent glutathione peroxidases (Se-GPx)), metallothioneins and lipid peroxidation (LPO) in the gills and mantle of the mussel Bathymodiolus azoricus from Menez–Gwen hydrothermal vent site was evaluated and related to the accumulated metal concentrations (Ag, Cu, Cd, Fe, Mn and Zn) in the tissues. Maximum antioxidant enzyme activities in the gills were detected in the beginning of summer, followed by a gradual decrease throughout the following months. One year after, the levels of antioxidant enzyme activities were similar to those reported one year before. LPO in this tissue exhibited a similar temporal variation trend. A different pattern of temporal variation in antioxidant enzyme activities was observed in the mantle, with a gradual increase from summer to the end of autumn (November). LPO in the mantle exhibited an almost reverse trend of temporal variation to that of antioxidant enzyme activities in this tissue. Antioxidant defences in the gills of B. azoricus were significantly enhanced with increasing concentrations of Ag, Cu and Mn, while negative relationships between antioxidant enzymes and Cd, Cu, Mn and Zn concentrations in the mantle were observed, suggesting different pathways of reactive oxygen species (ROS) production and that these tissues responded differently to the metal accumulation. However, temporal variation in biomarkers of defence and damage were in general similar to coastal bivalve species and can be associated with temporal variations of the physiological status due to reproduction. These variations might also be linked to the highly unstable nature of the hydrothermal environment.     

Metal concentrations in the shell of Bathymodiolus azoricus from contrasting hydrothermal vent fields on the Mid-Atlantic Ridge

Specimens of Bathymodiolus azoricus were sampled along the Mid-Atlantic Ridge at the Menez Gwen, Lucky Strike and Rainbow hydrothermal fields. Individual shells (n = 51), through the weight range 0.62 to 15.70 g, were analyzed for their magnesium, strontium, iron, manganese, copper, zinc and cadmium concentrations. Amongst the marine molluscs the shell of B. azoricus is confirmed as being particularly impoverished in strontium (mean 943 microg g(-1)). Trace metal concentrations in the shells decreased in the order Fe> Mn> Zn> Cu> Cd. Despite originating from trace metal rich environments mean concentrations were low (37.9, 13.2, 10.7, 1.1 and 0.7 microg g(-1), respectively). Irrespective of geographical origin magnesium, strontium and copper concentrations were primarily dictated by shell weight. In contrast cadmium concentrations were elevated in shells from the Rainbow field and ambient seawater chemistry imparted site specific chemical fingerprints to the shells with respect to the iron to manganese ratio.     

Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricus

Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se-GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 microM), Cu (0.4 microM) and Hg (0.1 microM) with different times of exposure. The experiments were performed in pressurized containers at 9+/-1 degrees C and 85 bars. Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.     

栉孔扇贝热休克蛋白70（HSP70）基因的BAC-FISH 定位

利用BAC-FISH技术,将包含HSP70基因的BAC克隆定位到栉孔扇贝的一对同源染色体的长臂上.HSP70基因的染色体定位将对深入研究该基因的结构及功能并将其应用于生产实践提供基础支持.同时,本研究是首次对栉孔扇贝低拷贝基因进行染色体定位的实践,其结果将为栉孔扇贝的染色体的深入研究以及染色体鉴别等工作提供必要的参考.     

Adaptation of the antioxidant defence system in hydrothermal-vent mussels (Bathymodiolus azoricus) transplanted between two Mid-Atlantic Ridge sites

The vent mussel Bathymodiolus azoricus is the dominant member of the Northern Mid‐Atlantic Ridge (MAR) hydrothermal megafauna, and lives in an environment characterized by temporal and spatial variations in the levels of heavy metals, methane and hydrogen sulphide, substances which are known to increase reactive oxygen species levels in the tissues of exposed organisms. To evaluate the effects of two contrasting hydrothermal environments on the antioxidant defence system of this vent mussel species, a 2‐week transplant experiment was carried out involving mussels collected from the relatively deep (2300 m), and chemical rich, Rainbow vent field. These were transplanted to the shallower (1700 m), and relatively less toxic, Lucky Strike vent field. To achieve this objective, levels of superoxide dismutase, catalase (CAT), total glutathione peroxidase (GPx), selenium‐dependent glutathione peroxidase and lipid peroxidation (LPO) were measured in the gills and mantle tissues of resident and transplant mussels before and after the transplant experiment. With the exception of CAT, the gills of the transplanted mussels had significantly higher antioxidant enzyme activity compared with the basal levels in the donor (Rainbow) and recipient (Lucky Strike) populations; whereas the antioxidant enzyme levels in the mantle tissues of the transplants reflected the baseline levels of activity in the native Lucky Strike mussels after 2 weeks. In contrast, LPO levels were significantly higher in both tissue types in the transplants than in either the source or the recipient populations, which suggested a response to hydrostatic pressure change (note, the transplant animals were brought to the surface for transportation between the two vent fields). The fact that the Rainbow mussels survived the transplant experience indicates that B. azoricus has a very robust constitution, which enables it to cope behaviourally, physiologically and genetically with the extreme conditions found in its naturally contaminated deep‐sea environment.     

荧光定量PCR方法分析皱纹盘鲍HSP70在温度胁迫下的表达

根据皱纹盘鲍(Haliotis discus hannaiIno)HSP70和Actin mRNA序列,设计合成引物,建立了皱纹盘鲍HSP70的荧光定量PCR技术平台,并用于检测皱纹盘鲍在热激处理后不同恢复时间HSP70的转录水平变化。结果表明在热诱导后,肌肉组织HSP70的表达呈时间依赖性,存在先升高后降低的趋势,在热诱导后2h,肌肉HSP70基因的表达量明显升高,是对照组(0h)的15倍,随后HSP70的表达量继续升高,在诱导后12h达到最高值,是对照组的21倍。然后表达量开始回落,至热诱导后96h,HSP70表达量降至初始值,表现出一种瞬时表达的趋势。     

Bottom fauna associated with <Emphasis Type="Italic">Bathymodiolus azoricus</Emphasis> (Mytilidae) mussel beds in the hydrothermal fields of the Mid-Atlantic Ridge

The structure of the assemblages associated with the mussel aggregations of Bathymodiolus azoricus was investigated. The mussel beds were found on the hydrothermal vent fields on the Mid-Atlantic Ridge (the Menez Gwen, Lucky Strike, and Rainbow areas) at the depths of 850–2400 m. The community structure of the mussel bed assemblages varied between the studied areas. Large number of species was unique to Menez Gwen mussel beds; the most observed taxa were not specialized hydrothermal species. All the other, nonunique species were found for the Lucky Strike region. The lowest mussel assemblage structure evenness was observed in the shallowest area, the Menez Gwen area (850 m depth). We assume that two types of mussel assemblages—nematode-dominated and copepod-dominated ones—exist in the Lucky Strike field. The assemblages of B. azoricus differ significantly from the assemblages of B. thermophilus inhabiting the Pacific hydrothermal vents.     

Growth increments and stable isotope variation in shells of the deep-sea hydrothermal vent bivalve mollusk Bathymodiolus brevior from the North Fiji Basin,Pacific Ocean

Bathymodiolus brevior [von Cosel, R., Métivier, B., Hashimoto, J., 1994. Three new species of Bathymodiolus (Bivalvia: Mytilidae) from hydrothermal vents in the Lau Basin and the North Fiji Basin, western Pacific, and the Snake Pit Area, mid-Atlantic ridge. Veliger 37, 374–392] a bivalve mollusk living at deep-sea hydrothermal vents, exhibits daily microgrowth structures in its shell. This interpretation is substantiated by various lines of evidence: (1) similar shell portions of contemporaneous specimens from the same locality contain almost the same number of microincrements; (2) the number of microincrements coincides with the expected number of days in which shell portions of Bathymodiolus spp. form; (3) the width of such microincrements compares well with daily growth rates estimated for the close relative B. thermophilus [Kenk, V.C., Wilson, B.R., 1985. A new mussel (Bivalvia, Mytilidae) from hydrothermal vents in the Galapagos rift-zone. Malacologia 26, 253–271]; (4) different specimens from the same site show similar microgrowth curves. In addition, we found support for tide-controlled shell growth. Daily shell growth rates fluctuate on a fortnightly basis. Some shell portions also revealed the typical tide-controlled microgrowth pattern commonly observed in intertidal bivalves. Based on the analyses of lunar daily growth increments, a growth curve for B. brevior was computed: X_t=14 cm−(14–0.04 cm) e^−0.26t. This curve enables estimation of ontogenetic age from shell length. According to this equation, B. brevior reaches its maximum shell length of 14 cm at about age 18. Shell isotope analyses suggest that some major shell growth interruptions or retardations are related to extremely active hydrothermal venting activity. However, shell growth also stopped during periods of low venting implying physiological controls on shell formation. Results of the present study demonstrate that shells of B. brevior provide calendars and environmental data loggers that can complement or partly substitute for long-term observations of venting systems.     

Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricus

Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se–GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 μM), Cu (0.4 μM) and Hg (0.1 μM) with different times of exposure. The experiments were performed in pressurized containers at 9 ± 1 °C and 85 bars.Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.     

Variations in deep-sea hydrothermal vent communities on the Mid-Atlantic Ridge near the Azores plateau

Near the Azores Triple Junction as the Azores Plateau is approached, the ridge axis becomes shallower; its depth decreases from ca. 2400 m in the Rainbow vent field (36°13′N) to ca. 850 m in the Menez Gwen vent field (37°35′N). In this area, extensive mussel beds of the mytilid Bathymodiolus azoricus dominate the hydrothermal vent fauna, along with populations of three shrimps (Rimicaris exoculata, Mirocaris fortunata and Chorocaris chacei). The main physical and chemical characteristics of the vent habitat were studied by discrete sampling, in situ analysis and sediment trap moorings. The vent fauna is distributed along a variable band where the vent fluids and seawater mix, with R. exoculata living in the most concentrated areas and Bathymodiolus azoricus in the most diluted zones. Various non-endemic species live at the border of the vent field. The variations observed in structure and composition of the communities along the depth gradient are most likely due to changes in vent fluid toxicity (metallic and sulphide content) and suspended mineral particles, which render the fluids harsher for species living there. The main faunal differences observed between Lucky Strike and Menez Gwen hydrothermal fields are due to an impoverishment in the hydrothermal endemic species and to the penetration of bathyal species. The comparison of the three studied vent fields suggests the existence of a succession of several biogeographic islands rather than a single province.     

Relationship between metal levels in the vent mussel Bathymodiolus azoricus and local microhabitat chemical characteristics of Eiffel Tower (Lucky Strike)

The turbulent mixing of hydrothermal hot fluid with cold seawater creates large chemical gradients at a small spatial scale that may induce variable physiological and biochemical adaptations within the vent fauna. The adaptation to such a variable environment by the vent mussel Bathymodiolus azoricus relies on a dual symbiosis hosted in the gills, and digestion of particulate organic matter. The surrounding environment not only provides the necessary energy sources and suspended organic particles for the vent mussel nutrition, but also potentially toxic compounds such as metals. Our main goal was to see if there is a relation between metal accumulation in mussel organs and the chemical characteristics of their close environment. Mussels were collected at six locations in a cold part of the Eiffel Tower fluid-seawater mixing zone, characterized by distinct chemical compositions. Metals (Cd, Cu, Fe and Zn) and metallothioneins were quantified in the gills and digestive gland. The physiological condition of the sampled mussels was also evaluated using tissues and gill indices. Our study indicates that the accumulation of metals in B. azoricus is related to their spatial distribution and linked to fine scale environmental conditions that influence the physiological status of the organism.     

Effects of PCBs sorbed to algal paste and sediments on the stress protein response (HSP70 family) in the eastern oyster, Crassostrea virginica.

This study examined the stress protein response (HSP70 family) of reproductively inactive oysters fed 0.7 g algal paste containing 0, 0.35 and 3.5 micrograms polychlorinated biphenyls (PCBs) daily. A second set of treatment groups investigated the combined effect of PCBs and sediments (0.3 g sediments daily per oyster) on HSP70 response. After 8 weeks of PCB exposure, oyster tissues (mantle and gill) were sampled and analyzed for HSP70. Preliminary results did not show a significant effect in HSP70 response in oysters fed PCB sorbed to algal paste, albeit PCBs accumulated up to 1342 ng/g dry weight in the mantle, and up to 180 ng/g dry weight in gill tissues. However, the addition of sediments caused a significant increase in HSP70 levels of gills and mantle, although the mantle was less sensitive to the sediments.     

台湾龟山岛东南部海域白色热泉特征分析

海底热液活动及其周边环境的调查研究是海洋科学领域的热点之一。海底热液喷口环境的物理化学参数,尤其是浅水型热液,会受到潮汐变化的影响。台湾龟山岛靠近冲绳海槽南端分布有多处浅水型热液喷口(30m)。2010年5月,浙江大学、杭州电子科技大学和台湾中山大学对该海域实施了联合考察,利用电化学传感器获得了低温热液喷口(24.834 14°N,121.961 91°E)连续87h的温度和pH现场观测数据。基于Morlet复小波变换方法,分析了浅水型热液喷口的温度和pH现场观测时间序列的变化周期,并通过与当地潮汐记录数据作比较,获得了其与潮汐信号之间的潜在关系,证实了潮汐变化是海底白色热泉喷口物理化学参数现场观测最主要的影响因素。温度和pH现场观测数据经Morlet复小波变换后的小波系数分别存在规律的12.9h和12.5h变化周期,并且温度的小波系数在准半日周期尺度与潮汐高度呈负相关变化,相关系数为-0.89;pH的小波系数情况相反,相关系数为0.76。     

克氏原螯虾热休克蛋白70 ku类似物基因的cDNA分析

根据已测序的克隆号PCI246基因序列设计2条引物,用快速扩增cDNA末端技术扩增克氏原螯虾(Procambarus clarkii)70 ku热休克蛋白同类物基因cDNA的5'端片段和3'端片段,测序得到1 184 bp基因片段,包含该基因的完整3'端,GenBank登录号AY357302,与GenBank序列号AB016836家蚕(Bombyx mori)的70 ku热休克蛋白同类物mRNA的部分片段83%同源。和果蝇(Drosophila melanogaster)的热休克蛋白同类物3的同源性最高(GenBank的登录号分别是NP_727 563,NP_511 132,NP_727 564,NP_727 565),为85%。     

深海热液区管状蠕虫Ridgeia piscesae中Ricin B-lectin型结构域基因的克隆与表达

作为一种重要的模式识别因子,凝集素在多个领域中均有着广泛的应用价值,因此,开发和利用新型凝集素将具有重要意义.本研究以生活在深海热液区极端环境条件下的管状蠕虫Ridgeia piscesae为材料,首次从管状蠕虫中克隆获得Ricin B-lectin基因rgal.序列分析表明,RGAL与已知序列的相似性较低,其含有2个Ricin B-lectin型结构域,并且该结构域具有该家族所特有的β-三叶草形三维结构.多方面的分析结果显示RGAL可能是一个新颖的凝集素蛋白.对RGAL进行了原核重组表达,并对其复性条件进行了摸索优化,成功获得了其可溶性表达产物,为后续RGAL的生物活性分析奠定了坚实的基础.     

Expression of heat shock protein 70 in a permanent cell line (EPC) exposed to sediment extracts from the North Sea and the Baltic Sea

Heat shock proteins 70 (hsp70) are known to be induced by a great variety of chemical stressors. The effects of different environmental contaminants, which were identified in sediments of the North Sea and the Baltic Sea as main contaminants or as contaminants of special toxicological interest, on the extent of expression of hsp70 in a permanent cell line Epithelioma papulosum cyprini (EPC) were investigated. Concentrations of the single contaminants leading to elevated hsp70 levels were compared to the respective concentrations of these contaminants encountered in the sediment extracts thus allowing an assessment of their contribution to the toxicity of these sediments. Analysis of hsp70 was performed using western blotting techniques with subsequent comparative quantification by densitometry. Eleven of thirteen contaminants have shown a dose-dependent increase in hsp70 levels. An expression of hsp70 was observed in most cases between 0.1 and 10 microg/mL test solution. Some of the investigated stressors led to extremely high hsp70 contents even at low concentrations. However, in most cases contaminant concentrations in sediments were too low to cause an effect. For this reason, effects of the tested sediments are attributed to other contaminants or rather to mixtures of stressors.     

Microbial community structure and diversity in deep-sea hydrothermal vent sediments along the Eastern Lau Spreading Centre

The aim of this study is to investigate microbial structures and diversities in five active hydrothermal fields’ sediments along the Eastern Lau Spreading Centre (ELSC) in the Lau Basin (southwest Pacific). Microbial communities were surveyed by denatured gradient gel electrophoresis (DGGE) and clone library analysis of 16S rRNA genes. The differences in microbial community structures among sediment samples from the five deep-sea hydrothermal sites were revealed by DGGE profiles. Cluster analysis of DGGE profiles separated the five hydrothermal samples into two groups. Four different 16S rRNA gene clone libraries, representing two selected hydrothermal samples (19-4TVG8 and 19-4TVG11), were constructed. Twenty-three and 32 phylotypes were identified from 166 and 160 bacterial clones respectively, including Proteobacteria, Bacteroidetes, Firmicutes, Nitrospirae and Planctomycetes. The phylum Proteobacteria is dominant in both bacterial libraries with a predominance of Gamma-Proteobacteria. A total of 31 and 25 phylotypes were obtained from 160 and 130 archaeal clones respectively, including Miscellaneous Crenarchaeotic Group, Marine Group Ⅰ and Ⅲ, Marine Benthic Group E, Terrestrial Hot Spring Crenarchaeota and Deep-sea Hydrothermal Vent Euryarchaeota. These results show a variety of clones related to those involved in sulfur cycling, suggesting that the cycling and utilization of sulfur compounds may extensively occur in the Lau Basin deep-sea hydrothermal ecosystem.