Linkages between cellular biomarker responses and reproductive success in oysters--Crassostrea virginica

The purpose of these studies was to evaluate if there were relationships between lysosomal destabilization or glutathione concentrations and gamete viability of oysters, Crassostrea virginica. Oysters were collected from field sites during the peak spawning period (May-June) during 2001 and 2002. Lysosomal destabilization rates and glutathione concentrations of hepatopancreas tissues (e.g. digestive gland) were determined. Eggs and sperm from the same adults were also used to conduct embryo development assays with reference seawater collected from a clean site, site water, and also a range of Cd concentrations (the Cd exposures were used to determine if there were differences in susceptibility to pollutants). Baseline embryo development success (e.g. percent normal development when the assays were conducted with reference seawater or site water) was related to lysosomal destabilization, but not glutathione status. However, the susceptibility of embryos to metal toxicity was related to glutathione status, i.e., sensitivity to Cd exposures increased with decreasing glutathione levels. These studies support the hypotheses that there are linkages between embryo development success and susceptibility to pollutant stress and cellular biomarker responses. These kinds of effects on reproductive success could lead to subtle but significant long-term effects on recruitment and viability of oyster populations.     

Seasonal variation in lysosomal destabilization in oysters,Crassostrea virginica

Lysosomal destabilization assays have been used as valuable biomarkers of pollutant exposures in a variety of bivalve and fish species. The responses of oysters, Crassostrea virginica, deployed at and native to various reference and degraded sites were evaluated for lysosomal destabilization during both summer and winter seasons. In both native and deployed oysters, lysosomal destabilization rates tended to be higher during the winter at both reference and polluted sites. There are at least two hypothetical explanations. Greater lysosomal destabilization rates may be related to physiological changes associated with mobilization of nutrient reserves during the winter and gametogenesis. However, lysosomal destabilization in deployed oysters was correlated with tissue metal concentrations. These data also support a second hypothesis that seasonal differences in physico-chemical factors (such as reduced levels of acid volatile sulfides) may increase the bioavailability of metals during the winter so that adverse effects are more pronounced.     

The effects of glutathione depletion on reproductive success in oysters, Crassostrea virginica.

Glutathione (GSH) is a ubiquitous tripeptide that functions as a very important modulator of cellular homeostasis, including detoxification of metals and oxyradicals. Therefore, depletion of GSH may predispose organisms to pollutant stress. Reproductively active oysters (Crassostrea virginica) were exposed to buthionine sulfoximine in the laboratory to deplete gonadal GSH. The effects of metal exposures (Cd and Cu) on fertilization and developmental assays were evaluated using gametes from control and GSH-depleted adults. Fertilization success was not affected by GSH status, i.e. the fertilization rates of gametes derived from GSH-depleted adults were the same or slightly higher. However, GSH depletion did increase the susceptibility of developing embryos to metal toxicity, i.e. adverse effects on embryonic development were observed at lower metal concentrations with gametes derived from GSH-depleted adults. These effects may be related to diminished removal of free radicals or increased availability of metals. Whereas sperm penetration of embryonic membranes and fertilization success may be facilitated by free radicals, the persistence of free radicals during subsequent developmental periods may adversely affect differentiation and normal development. GSH probably also plays an important role in scavenging toxic metals and reducing metal interactions with essential developmental processes. These results suggest that parental depletion of GSH may increase the susceptibility of embryos to metal toxicity.     

An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, for monitoring the impact of an offshore platform in the Adriatic Sea

An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, was developed to evaluate the potential impact of an offshore gas platform in the central Adriatic Sea. Reference organisms were collected on a seasonal basis from an unpolluted site and transplanted for four weeks in both the sampling area and to the investigated platform. Chemical analyses of trace metals in mussel tissues were integrated with a multi-biomarker approach for the early detection of biological responses at several cellular targets. Induction of metallothioneins, peroxisomal proliferation and activity of acetylcholinesterase were measured as markers for specific classes of chemicals. Special attention was given to oxyradical metabolism and appearance of oxidative-mediated toxicity to reveal a more general onset of cellular disturbance. In addition to individual antioxidants (superoxide dismutase, catalase, glutathione S-transferases, glutathione reductase, Se-dependent and Se-independent glutathione peroxidases, and levels of total glutathione), the total oxyradical scavenging capacity (TOSC) allowed a quantification of the overall capability to neutralize specific forms of intracellular reactive oxygen species (ROS; i.e. peroxyl and hydroxyl radicals). Cellular damages were evaluated as lysosomal destabilization (membrane stability, accumulation of lipofuscin and neutral lipids), lipid peroxidation products (malondialdehyde) and DNA integrity (strand breaks and micronuclei); the air survival test was finally applied to evaluate the overall physiological condition of mussels. Concentration of trace metals (As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Zn) revealed only limited variations in transplanted mussels during various experimental periods and such changes appeared partly related to natural fluctuations. Among biological responses, variations of antioxidants and lysosomal stability were confirmed as sensitive early warning signals for biological disturbance of both natural and anthropogenic origin. The presented protocol with caged mussels allowed marked biological effects caused by the investigated platform to be excluded, and represented a useful approach that is easy to extend for monitoring the impact of offshore activities in the Adriatic sea.     

Sea urchin sperm bioassay for sewage and chlorinated seawater and its relation to fish bioassays

Bioassays were conducted with sea urchin and sand dollar sperm to determine the toxicity of chlorinated and unchlorinated sewage effluent and chlorinated and brominated seawater. The sperm cells were exposed to seawater dilutions of each toxicant for 5–15 min. The fertilisation of eggs served as the indicators of sperm viability. The effective concentrations which reduced fertilisation success by 50% (EC50) averaged 2·2 and 4·8% chlorinated and unchlorinated sewage in seawater, respectively. The sperm cells were extremely sensitive to chlorinated seawater at concentrations from 0·002 to 0·020 mg/litre total residual oxidant (TRO). Brominated seawater proved toxic to sperm in one test at 0·015 mg/litre TRO. Results of the sperm bioassays are compared with previous acute and chronic bioassays with fish.     

Behavior of Cu, Ni and Cd during nutrient depletion in a spring bloom in Funka Bay

The concentrations of Cu, Ni and Cd were determined in Funka Bay during a spring phytoplankton bloom, consisting of diatoms. Just after the bloom, both dissolved Cd and nutrients were removed in the euphotic zone. However, the removal ratio of Cd to phosphate was very different from that in seawater. The removal of Cd took place at a Cd/phosphate ratio of 0.07×10⁻³, which was lower than in seawater before the bloom (0.25×10⁻³), leading to an increase in this ratio in seawater exceeding 0.7×10⁻³ at the end of the bloom. Elevated concentrations of Cd and phosphate were observed in the deeper layer after the bloom due to the decomposition of detrital materials produced in the bloom. The ratio of Cd/phosphate in the regeneration step was 0.24×10⁻³ which was different from the removal ratio of 0.07×10⁻³. These observations suggest that the high Cd/phosphate ratio in the regeneration would reflect a relatively high regeneration rate of Cd than that of phosphate. No significant decrease in Cu and Ni concentrations was observed during the development of the bloom, suggesting that biological removal of these metals was not so significant during the spring bloom. The concentrations of Cd, Cu and silicate in surface waters increased after the bloom with decreasing salinity due to the influence of a spring thaw.     

Significance of metallothioneins and lysosomes in cadmium toxicity and homeostasis in the digestive gland cells of mussels exposed to the metal in presence or absence of phenanthrene

Metallothioneins and lysosomes are known to be involved in cellular detoxication and sequestration of certain metals^1–3 and both have been identified in this role in elimination of copper from marine mussels (Mytilus edulis/galloprovincialis).³ Cadmium (Cd), however, has been shown to persist in the cells of the digestive gland for long periods with only minimal elimination. An experiment was designed to test the effects of Cd on the fragility of lysosomal membranes in the digestive cells as a measure of cellular injury,^4,5 metallothionein content of the digestive gland and cadmium concentration in this organ. Phenanthrene was used also to destabilise lysosomal membranes⁶ in order to test if increased lysosomal fragility interfered with cadmium metabolism and detoxication. The results demonstrated that Cd induced metallothionein synthesis and that elimination of Cd was minimal after 28 days in clear seawater. Lysosomal fragility was initially increased but this effect was soon reversed, even with continued exposure to Cd. The lysosomal destabiliser, phenanthrene, did not appear to affect accumulation of Cd or levels of metallothionein.     

The distribution and preferential biological uptake of cadmium at 6°W in the Southern Ocean

The spatial and temporal distribution of cadmium (Cd) and phosphate in the Southern Ocean are related to biology and hydrography. During a period of 18 days between transects 5/6 and 11, a phytoplankton spring bloom developed in the Polar Frontal region. Upper water Cd concentrations were not depleted and ranged from 0.2 to 0.8 nM at about 10 m depth. These relatively high Cd concentrations are attributed to upwelling of Upper Circumpolar Deep Water (0.5–1.2 nM in the core) in combination with low biological productivity (0.2 to 0.3 mg m⁻³ chlorophyll-a, 0.3 g C m⁻² d⁻¹). Total particulate Cd concentrations at 40 m depth were between 0.02 and 0.14 nM with the maximum in concentration in the Polar Frontal region. Most of the particulate Cd at this depth (85–94%) was detected in the first phase of a sequential chemical leaching treatment which includes adsorbed Cd as well as Cd incorporated in algae. The Polar Frontal region was characterized by minima in Cd concentration and Cd/phosphate ratio of seawater at both transects; values were the lowest at transect 11 after development of the spring bloom which was dominated by diatoms. This decreasing Cd/phosphate ratio in seawater during spring bloom development was attributed to preferential Cd gross uptake which more than compensated the process of preferential Cd recycling. Within the Upper Circumpolar Deep Water, Cd showed a maximum in concentration similar to that of the major nutrients. Both the Cd concentration and the Cd/phosphate ratio of the deeper water increased in southern direction, from 0.4 to 0.7 nM and from 0.2 to 0.3 nM/μM, respectively. Antarctic Intermediate Water has a Cd concentration of 0.21 nM with a Cd/phosphate ratio of 0.10 nM/μM. In Antarctic Bottom Water, Cd concentrations ranged from 0.60 to 0.82 nM.     

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.     

Stability of lysosomal and cell membranes in haemocytes of the common mussel (Mytilus edulis): effect of low temperatures.

Expanding industrial activities in the Arctic require an urgent assessment of the toxicity of chemicals at low temperatures. Organisms acclimatized to low temperature exhibit specific adaptations. For example, the amount of unsaturated lipids is increased to maintain the fluidity of the cell membranes. It has been hypothesized that such temperature-induced alterations in membrane lipid composition may affect the stability of lysosomal and cell membranes in the common mussel, Mytilus edulis, an organism exposed to seasonal temperature extremes. As mussels may be exposed to petroleum compounds along industrialized coastlines, we tested the combined effects of exposure to low temperature and the petroleum compound, phenanthrene, on haemocyte membrane stability. Test animals, acclimated to either 0 or 10 degrees C, were exposed to phenanthrene (0 = control or 500 micrograms l-1) and haemocytes were examined using the neutral red retention assay (lysosomal stability) and a fluorescence assay (cell membrane stability). At 0 degree C, lysosomal and cell membranes from uncontaminated mussels were destabilized compared with 10 degrees C (P = 0.0005). No significant effects (P > 0.05) of phenanthrene were detected at either temperature. Possible mechanisms underlying membrane destabilization include a weaker physical resistance of the membrane due to a higher amount of unsaturated lipids, a potentially higher level of reactive oxygen radicals at low temperature and the higher susceptibility of unsaturated lipids to oxidative stress. More work is required to better understand the consequences of this membrane destabilization at low temperature on the susceptibility of the organism to pollutants.     

大亚湾珊瑚礁痕量金属的研究

作者采用阳极溶出伏安法测定大亚湾珊瑚礁痕量金属铜、铅、镉的含量 ,并讨论了相关的影响因素。测定结果发现 :A组、B组珊瑚礁礁体中铜、铅、镉的含量从里到外有递增的趋势 ,这与近年来大亚湾水体中铜、铅、镉含量的变化是一致的。而 A组珊瑚礁变化明显 ;B组珊瑚礁礁体中铜、铅、镉含量呈现较复杂的变化。分析认为 ,这是因珊瑚礁的形成受多种因素影响所致 :(1)海水组成对珊瑚礁化学成分的影响 ;(2 )海洋生物种类、数量及其矿物组成是制约元素分布的主要因素 ;(3)陆源碎屑组分对大亚湾珊瑚礁的化学组成有一定的影响。各因素在不同情况下所起的作用不同。     

The effects of silver nanoparticles on oyster embryos

Nanoparticles may be introduced into aquatic environments during production processes and also as a result of release following their use in various commercial formulations and biologic applications. Filter-feeding bivalve mollusks such as oysters are valuable model species for characterizing nanoparticle bioavailability and interactions with basic cellular processes. The adults release their gametes into the environment, so their embryos and larvae are also likely targets of nanoparticles. The purpose of these studies was to characterize the toxicity of metal nanoparticles on embryonic development of oysters, Crassostrea virginica and to compare the relative sensitivity of embryos to adults. Newly-fertilized oyster embryos were exposed to silver nanoparticles (AgNP) and then the percent normal development after 48 h was assessed. Studies were conducted with adult oysters in which they were also exposed to AgNP for 48 h, and the effects on lysosomal destabilization were determined. The expression of metallothionein (MT) gene expression was also assessed in both embryos and adults. Adverse effects on embryonic development were observed at concentrations similar to those that caused both statistically and biologically significant effects on lysosomal destabilization of adults. Significant increases in MT mRNA levels were observed in both embryos and adult oysters, and MT levels were highly induced in embryos. While we do not know whether the toxicity and gene expression responses observed in this study were due to the nanoparticles themselves or the Ag ions that dissociated from the nanoparticles, these kinds of basic studies are essential for addressing the potential impacts of nanoengineered particles on fundamental cellular processes as well as aquatic organisms.     

Cadmium bioconcentration in the echinoid Paracentrotus lividus: Influence of the cadmium concentration in seawater

Uptake and loss kinetics of cadmium were studied in Paracentrotus lividus exposed for 24 d to different stable Cd concentrations (0 to 1.14 μg Cd l⁻¹) and ¹⁰⁹Cd tracer added to seawater. The whole-body uptake kinetics were linear over the time course of the experiment. The whole-body ¹⁰⁹Cd concentration factor was independent of the stable Cd concentration in ambient seawater. Bioconcentration of Cd in whole individuals and their body compartments was directly proportional to the Cd concentration added to the seawater. The echinoid digestive tract wall showed the highest degree of Cd uptake. Whole-body loss kinetics were described by a 2-component exponential equation. The loss kinetics were similar for each of the treatments examined. Cadmium was found to display a long biological half-life in echinoid tissues; the major fraction (73 to 85%) of the Cd taken up by echinoids was eliminated following a loss component whose biological half-life was of the order of 1 year, regardless of the absolute Cd concentration accumulated by the echinoid. Paracentrotus lividus is suggested as a valuable biomonitor of Cd, particularly where knowledge of Cd concentration variations over long-term is of interest.     

Zinc concentration effect at the organismal,cellular and subcellular levels in the eastern oyster

The purpose of this research was to demonstrate a concentration effect of zinc exposure at organismal, cellular and sub-cellular levels in the eastern oyster and to find associated protein expression signatures (PES) for each concentration of zinc. Oysters were exposed to six concentrations of zinc for 48 h in a controlled environment. At the organismal level, fecal material was observed as a measure of physiological health during metal exposures. At the cellular level, lysosomal destabilization was measured using hemolymph. This cellular response was significant only at the highest concentration, when the fecal index was lowest. Protein responses were monitored in the oyster following exposure to zinc. Gill tissue was excised and homogenized, and then analyzed using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and digital image analysis. Protein expression signatures were found to be specific to each concentration. The protein responses were linked to the other biological parameters measured, each of which followed a concentration gradient of zinc.     

经济海藻羊栖菜（Sargassum fusiforme）幼体对镉胁迫的响应特性

镉(Cd)是水生生态环境中常见的重金属污染物,通常会抑制植物的生长,减少生物量,对植物产生毒害作用。为了研究羊栖菜幼苗对海水中Cd胁迫的抗氧化响应特性,我们在天然海水和Cd加富(100 μmol·L^-1)海水两种不同的Cd浓度下培养羊栖菜幼体。实验结果表明:Cd加富胁迫显著抑制了羊栖菜幼苗生长,使藻体颜色变暗。此外,高Cd处理显著降低了羊栖菜中的色素含量,生长速率,过氧化物酶(POD)活性,脱氢抗坏血酸(DHA)含量和谷胱甘肽还原酶(GR)活性。相反,高Cd处理显著促进了藻体内的Cd元素积累、对Cd₂⁺吸收速率,并增加了其暗呼吸/净光合速率比值(R_d/P_n),抗坏血酸(Vc)含量,可溶性蛋白质(SP)含量,且还原型谷胱甘肽(GSH)以及羊栖菜的超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性均显著增加,但对丙二醛(MDA)的影响不显著。虽然羊栖菜幼苗增加了其抗氧化活性并促进AsA-GSH循环,以产生H₂O₂并维持健康的新陈代谢,但高Cd胁迫显著限制了幼苗的光合作用,降低了其光合色素,抑制了其生长,并使其藻体颜色发生变化。海水中高浓度的Cd对羊栖菜幼苗具有毒性作用,并且极大地增加了这种海藻的食用风险。     

Distribution Characteristics of Heavy Metals and Grain Size of Sediments from Hailing Bay, China

Introduction Hailing Bay located in western Guangdong province is an important marine aquaculture base and a famous scenic spot. Moyang River which flows across Yangjiang is the major runoff and polluted source in the sea area of Hailing Bay. Hailing Bank ( approximately 5 000 m ) which was built to connect Hailing Island and the continent in 1964 has blocked the natural circulation of suspended sand and pollutants in seawater. Water flux through the Yangjiang harbor extensively decreases …     

海水酸化和磺胺甲恶唑复合暴露对大马蹄螺抗氧化酶活性和脂质过氧化的影响

为探讨海水酸化和磺胺甲恶唑环境残留对珊瑚礁区生物的影响,本文以南海珊瑚礁区关键礁栖生物大马蹄螺（Trochus niloticus） 为研究对象,探究了两种环境胁迫因素对大马蹄螺的氧化胁迫作用。分别采用两个 pH 水平 （pH 8.1、 pH 7.6） 和三个磺胺甲恶唑浓度水平 （0 ng/L、100 ng/L 和 1 000 ng/L） 单独和复合暴露 28 天,测定其对大马蹄螺鳃组织的超氧化物歧化酶 （SOD）、过氧化氢酶 （CAT） 和谷胱甘肽巯基转移酶 （GST） 活性以及谷胱甘肽 （GSH） 含量和脂质过氧化水平 （LPO） 的影响。结果发现：海水酸化和磺胺甲恶唑单独暴露能够导致大马蹄螺鳃组织出现不同程度的氧化应激现象,表现为抗氧化酶 （CAT、SOD 和 GST） 活性和谷胱甘肽含量的升高。在复合暴露条件下,大马蹄螺鳃组织的抗氧化酶（CAT、SOD 和 GST） 活性和谷胱甘肽含量均受到不同程度的抑制,同时伴随着脂质过氧化水平的显著升高。这表明短期海水酸化和磺胺甲恶唑复合暴露可导致大马蹄螺鳃组织出现氧化损伤现象,进而可能对个体存活和种群繁衍造成负面影响。     

Accumulation of Zn,Cu and Cd by crabs and barnacles

The crab Carcinus maenas (L.) and the barnacle Elminius modestus Darwin were exposed to a range of dissolved concentrations of Zn, Cu and Cd for 21 days in artificial seawater. Accumulation of Zn and Cu by crabs has been interpreted in terms of the presence of a regulation mechanism to maintain constant body concentrations (83·2 ± 19·4 μg Zn g^?1 dry wt.; 39·8 ± 9·8 μg Cu g^?1 dry wt.) under varying external dissolved metal levels, until a threshold dissolved metal concentration (c. 400 μg Zn l^?1; c. 170 μg Cu l^?1) beyond which net accumulation of metal begins. Cadium appears to be accumulated by C. maenas at all exposures with no evidence for regulation of body cadmium concentrations. Exposure of E. modestus to Zn, Cu or Cd caused net accumulation of the respective metal in the bodies of the barnacles, with no evidence for regulation of body metal concentrations.     

Characteristics of the ratio of dissolved cadmium to phosphate in subtropical coastal waters of Ishigaki Island,Okinawa, Japan

The ratio of dissolved cadmium (Cd) to phosphate (PO₄) in the subtropical coastal area of Ishigaki Island, Okinawa, Japan, was investigated. Twenty vertical seawater samplings were carried out once a month from May 2008 to January 2010. In order to examine how the Cd/PO₄ ratio in seawater varies with the oceanographic conditions (i.e., the water temperature–salinity characteristics), the water masses at the study sites were classified into two types: group 1 with a water temperature of >25°C and a salinity range of 34.0–34.4, and group 2 with a water temperature of <25°C and a salinity of >34.4. A different phytoplankton assemblage was observed in each water mass defined. Different Cd/PO₄ ratios were obtained for the two water mass types, due to the differences between the types in terms of the environmental conditions such as the water temperature–salinity (T–S) characteristics and phytoplankton assemblages, as well as possible variations in the concentrations of dissolved iron, zinc, manganese, and CO₂ in seawater in each water mass.     

Effect of exposure time on the bioaccumulation of Cd, Mg, Mn and Zn in Cystoseira abies-marina samples subject to shallow water hydrothermal activity in São Miguel (Azores)

Shallow water hydrothermal vents can be compared to polluted places due to high concentrations of heavy metals, and are thus good models for bioaccumulation studies. The present study intended to estimate the time of exposure required for the accumulation of certain elements to stabilize in specimens of Cystoseira abies-marina , to be used as a reference in future work. Cystoseira abies-marina intertidal specimens were transplanted from Mosteiros (a non-hydrothermal and pristine site) to Ferraria (with hydrothermal activity) and left there. Transplanted samples were collected after 1, 2, 4 and 8 weeks and the concentrations of Cd, Mg, Mn and Zn were measured through flame atomic absorption spectrophotometry. Although further studies with increased periods of exposure are needed because the concentration of these elements never stabilized in the collected samples, there is strong evidence that increased time of exposure led to increased concentrations of Cd, Mg, Mn, but not Zn. These results are consistent with the assumption that C. abies-marina is bioaccumulating some of the heavy metals and can thus be a good indicator for polluted waters.