基于综合生物标志物响应指数法评估近岸海洋环境压力——以广西西部沿海为例

2011-2012年,在广西西部沿海设置5个站点,以野生文蛤作为指示生物进行3次采样监测.从个体、细胞和分子水平选择了钻沙所需时间、吞噬能力、溶酶体膜稳定性、血浆三价铁还原力、乙酰胆碱酯酶活性及彗星率6项指标,利用综合生物标志物响应指数模型(IBR),整合上述生物标志物指标,并转化为形象直观的星状图,对调查站点的环境状况进行风险评估.结果表明: 各站点的综合生物标志物响应指数值(IBR/n)介于2.30~8.68,茅尾海的环境压力最大,北仑河口最小.虽然不同的生物标志物对污染压力的响应存在差异,但利用IBR可以有效区分文蛤所在区域的环境压力状况.利用生物标志物的监测结果与化学监测结果基本吻合.     

Assessing environmental conditions of the Río Champotón (México) using diverse indices and biomarkers in the fish Astyanax aeneus (Günther, 1860)

The upper reaches of the Río Champotón in southwestern Mexico are largely unknown to science. This river is sustaining anthropogenic and natural disturbances by deforestation, substitution of natural vegetation, and agricultural non-point source pollution. Furthermore, hurricanes provoke flooding of these areas, exposing the aquatic biota to pollutants. Several approaches were used to assess the effects of environmental conditions along the freshwater portion of the Río Champotón in the native fish Astyanax aeneus and to determine the feasibility of extrapolating responses between levels of organization, and to link spatial and seasonal fish responses with water quality features. A water quality index (WQI) was employed as an indicator of environmental conditions, a set of sub-organismal biomarkers in A. aeneus (lipid peroxidation, glutathione S-transferase, ethoxyresorufin-O-deethylase and lactate dehydrogenase) was monitored to determine the integrated biomarker response (IBR), and somatic indices (individual and population-level responses; gonadosomatic, hepatosomatic, and condition factor) were characterized. Canonical correspondence analysis revealed spatial and seasonal variations in all the approaches. The dry and the wet season (April and July) exhibited low WQI scores, but decreased hardness and conductivity values as well as increased WQI values occurred during the post-hurricane season when crop fields adjacent to the riverbed were inundated. Although the WQI suggested low levels of contamination, the biomarkers did not remain static; wide fluctuations reflected fish exposure to seasonal stressors that provoked impairments in biological responses. All biomarker values were high, mainly in the post-hurricane season (November). The IBR exhibited two peaks that may be related to stress periods (April and November), with decreasing post-stress values suggesting the existence of homeostatic processes in the fish. The somatic indices reflected some seasonal and spatial differences at the population level – the gonadosomatic and hepatosomatic indices peaked in the middle of July and November, respectively – but the condition factor remained constant among sites and study periods. Future studies are needed to explore the link between the fish biomarker responses and possible causal stressors (natural or anthropogenic).     

Biochemical and toxicopathic biomarkers assessed in smallmouth bass recovered from a polychlorinated biphenyl-contaminated river

Smallmouth bass (Micropterus dolomieu) were collected to quantify the nature and prevalence of biomarker responses, including biochemical indices, toxicopathic lesions and general health indices, among fish collected from polychlorinated biphenyl (PCB)-contaminated and nearby uncontaminated reaches of the Kalamazoo River, Michigan, USA. Blood and tissue samples (gill, liver, spleen, head kidney, trunk kidney, thyroid and gonads) were collected and preserved at necropsy for biochemical and histological analyses. The body condition factor and liver somatic index were significantly lower in fish collected from the downstream, contaminated site. Plasma vitellogenin was not detected in male fish collected from either site. Liver ethoxyresorufin-O-deethylase activity and liver and spleen superoxide dismutase activity were significantly depressed in fish collected from the downstream site. Significant toxicopathic lesions such as glycogen depletion, enhanced macrophage aggregates, hepatic foci of cellular alteration (i.e. preneoplastic lesions) and neoplasia were also detected in the liver of fish collected from the downstream site. This study indicates that many of the biochemical and histopathological biomarker responses were associated with liver and body tissue PCB concentrations. Taken together, the biomarkers of exposure and effect strongly suggest that fish within the downstream site are adversely affected by PCBs and other chemical stressors.     

Biochemical and toxicopathic biomarkers assessed in smallmouth bass recovered from a polychlorinated biphenyl-contaminated river

Smallmouth bass (Micropterus dolomieu) were collected to quantify the nature and prevalence of biomarker responses, including biochemical indices, toxicopathic lesions and general health indices, among fish collected from polychlorinated biphenyl (PCB)-contaminated and nearby uncontaminated reaches of the Kalamazoo River, Michigan, USA. Blood and tissue samples (gill, liver, spleen, head kidney, trunk kidney, thyroid and gonads) were collected and preserved at necropsy for biochemical and histological analyses. The body condition factor and liver somatic index were significantly lower in fish collected from the downstream, contaminated site. Plasma vitellogenin was not detected in male fish collected from either site. Liver ethoxyresorufin-O-deethylase activity and liver and spleen superoxide dismutase activity were significantly depressed in fish collected from the downstream site. Significant toxicopathic lesions such as glycogen depletion, enhanced macrophage aggregates, hepatic foci of cellular alteration (i.e. preneoplastic lesions) and neoplasia were also detected in the liver of fish collected from the downstream site. This study indicates that many of the biochemical and histopathological biomarker responses were associated with liver and body tissue PCB concentrations. Taken together, the biomarkers of exposure and effect strongly suggest that fish within the downstream site are adversely affected by PCBs and other chemical stressors.     

Condition factor and hepatosomatic index as estimates of energy status in male three-spined stickleback

Seasonal variation in direct and indirect measures of energy status was examined using estimates of glycogen, lipid and protein levels in a single cohort of male three-spined sticklebacks from an annual population collected each month over one complete year. Condition factor, somatic condition factor and hepatosomatic index (HSI) were calculated as indirect indices of energy status and the accuracy of these indirect measures as predictors of energy status was investigated. Results indicate that both condition factors were significant predictors of energy reserves (lipid, protein, glycogen and total energy), but that the proportion of variance accounted for was small. Both condition factors perform better as predictors of energy content per unit body weight. The HSI was a significant, but a weak predictor of total glycogen levels over the whole year. On a seasonal basis the relationship between HSI and energy reserves was highly variable. These indices are therefore poor predictors of energy reserves in male three-spined sticklebacks.     

花色苷提取物对大鼠发育关键期铅致肝脏和肾脏损伤的保护作用研究

为研究天然色素花色苷（anthocyanins,ACNs）对铅中毒引起的脏器损伤的保护作用,构建了铅暴露大鼠模型,灌服不同剂量的ACNs溶液和强力排铅药二巯基丁二酸（dithioglysuccinic acid, DMSA）,连续3周,于末次给药24 h后考察脏器中的铅含量、组织病理学、血液及脏器生化指标;并采用综合生物标志物响应（integrated biomarker responses, IBR）评估ACNs对铅致发育期大鼠肝脏和肾脏氧化损伤的修复能力。实验结果表明,铅大量存在于大鼠的肝脏和肾脏组织中,造成脏器病理学结构及氧化损伤;而ACNs可促进铅排出机体,改善组织病理学结构,使血清中转氨酶、肌酐等生化指标水平明显好转,并升高肝和肾抗氧化酶的活性,减少还原性物质的含量,改善铅诱导组织的氧化损伤;IBR分析结果显示,ACNs可使铅损伤的脏器得到明显的修复。结果表明,ACNs营养干预可有效拮抗铅致机体的氧化损伤,从而有效修复铅损伤的肝肾组织。     

Development of a biomarker-based index for assessing the ecotoxic potential of aquatic sites

The use of biochemical or physiological measurements as indicators of ecotoxicity is under constant development and has the advantage of delineating effects before the appearance of disease. However, these biomarkers are often part of a battery of tests, and it is difficult to integrate them together to gain an overall view of an organism's health. The aim of this study was to develop an index that could integrate the data derived from a battery of biomarkers for application to both spatial and temporal studies. Mya arenaria clams were collected at different sites along the Saguenay Fjord (Quebec, Canada). Six biomarkers were measured: metallothioneins, DNA strand breakage, lipid peroxidation, vitellin-like proteins, phagocytosis, and non-specific esterase activity in haemocytes. A biomarker index was obtained by summing the biomarker values expressed in term of classes. Classes were determined by a distribution-free approach derived from the theory of rough sets. The results of the spatial study show that the index values discriminated well between contaminated and uncontaminated sites. The highly polluted sites had the highest index values (18 compared with a reference value of 14). In the temporal study, the index was also able to highlight possible contamination-induced alterations, even though the interpretation of temporal variation is complicated by natural variations occurring throughout the year. A control chart approach is proposed for determining contaminated sites in both spatial and temporal surveys.     

Influence of Molting and Starvation on Digestive Enzyme Activities and Energy Storage in Gammarus fossarum

Among the many biological responses studied in ecotoxicology, energy-based biomarkers such as digestive enzyme activities and energy reserves appear to be useful predictive tools for detecting physiological disturbances in organisms. However, the use of these biological responses as biomarkers could be limited by the effects of confounding factors (biotic and abiotic) and physiological processes, such as the reproductive cycle. Thus, the optimal use of these biomarkers will be facilitated by understanding the effects of these factors on the energy metabolism of the sentinel species being studied. We considered abiotic factors (temperature and conductivity) in a previous study, whereas the present study investigated the effects of gender, the female reproductive stage, and food availability on the digestive enzyme activities and energy storage of Gammarus fossarum. The results indicated that, during the female reproductive cycle, the activities of digestive enzymes (amylase, cellulase, and trypsin) decreased significantly, whereas the levels of reserves (proteins, lipids, and sugar) increased until the last premolt stage. Restricted food diets only led to decreased amylase activities in both sexes. Food starvation also induced a decrease in the energy outcomes in females, whereas there were no effects in males. In general, the biochemical (digestive enzyme activities) and physiological (energy reserves) responses were more stable in males than in females. These results support the use of males fed ad libitum to limit the effects of confounding factors when using these energy biomarkers in Gammarus fossarum during biomonitoring programs.     

Are biochemical biomarker responses related to physiological performance of juvenile sea bass (Dicentrarchus labrax) and turbot (Scophthalmus maximus) caged in a polluted harbour?

Biomarker responses to toxic exposure have been used for decades to indicate stress in aquatic organisms, or the magnitude of environmental pollution. However, little has been done to compare the simultaneous responses of both biochemical and physiological biomarkers. The purpose of this study was twofold. Firstly to analyse the responses of several biochemical biomarkers measured on juvenile sea bass and turbot caged in a northern France harbour at a reference and contaminated stations. Several biotransformation parameters (Ethoxyresorufin-O-deethylase - EROD - and Glutathione S-transferase -GST) and an antioxidant enzyme (Catalase -CAT) were analysed. Secondly, to compare their responses to several growth and condition indices, measured on the same fish. In the contaminated station, EROD and GST activities were found to be significantly higher, and a decrease of CAT activity was observed for both species. For individual sea bass, biochemical biomarkers showed numerous significant correlations with growth and condition indices, such as the Fulton's K condition index, the RNA:DNA ratio and the lipid storage index. On the contrary, there were only a few significant correlations for turbot, suggesting a species-specific response. Our study indicates that the analysis of the simultaneous responses of both biochemical and physiological biomarkers can be useful for monitoring complex exposure and to assess habitat quality.     

Seasonal variability in biomarkers in the bivalves Mytilus edulis and Macoma balthica from the northern Baltic Sea

Metallothionein level (MT), and acetylcholinesterase (AChE), catalase (CAT) and glutathione-S-transferase (GST) enzyme activities in the bivalves Mytilus edulis and Macoma balthica were investigated for seasonal variations from an inshore and an offshore site in the northern Baltic Sea. All the biomarkers showed variability, following mostly a similar pattern at both sites. Relationships between biomarkers and environmental factors and protein concentration and weight of target tissues were examined. In M. edulis, GST activity was related to Secchi depth, while in M. balthica a correlation with near-bottom oxygen saturation was observed. AChE activity correlated with the weight of the foot tissue of M. balthica. In both species, an integrated biomarker index indicated a stressed condition during the spring/early summer period. Strong seasonal variability in temperature and a concentrated period of food availability in spring-both governing the reproductive cycle of the bivalves-probably explains most of the observed natural variability in biomarkers in this sea area.     

Xenobiotic metabolism in bivalves

Xenobiotic metabolism is the aim of many studies concerning the development of pollution biomarkers. Enzymatic pathways of this metabolism in bivalves are similar to those previously described in vertebrates, each of them showing their own biochemical characteristics and sometimes specific mechanisms. Laboratory or in-situ induction experiments frequently lead to delicate interpretations. The analysis of these results displays a tendency for the induction of P-450 activities by hydrocarbons in marine bivalves, and a proportional induction of xenobiotic membranar elimination mechanisms to the level of in-situ pollution in Mytilus galloprovincialis. This synthesis confirms the possible use of enzymatic activities from xenobiotic metabolism as pollution biomarkers in bivalves. The lack of fundamental data and the low number of in-situ studies have prevented, until now, the establishment of a systematic correlation between an enzymatic behaviour and a level of pollution.     

基于生物标志物指数法的海洋环境评价方法综述

生物标志物对化学污染物具有“早期预警”功能,在海洋环境评价中应用广泛.以生物标志物为基础的综合指数能够整合多种标志物对环境状况的响应,因而成为评价环境质量的有用工具.这些综合指数方法包括多生物标志物污染指数(MPI)、综合生物标志物响应指数(IBR)、生物效应评价指数(BAI)、生物标志物响应指数(BRI)、健康状态指数(HIS)等.本文从生物标志物指标体系确定、综合指数计算方法、污染程度分级、应用效果等方面对这些评价方法进行综述,并对基于生物标志物指数的海洋环境评价方法存在的问题和改进建议进行了探讨.     

Toxicological effects of methylmercury on walleye (Sander vitreus) and perch (Perca flavescens) from lakes of the boreal forest

Biochemical and physiological responses of walleye (Sander vitreus) and perch (Perca flavescens) were studied in four Canadian boreal forest lakes representing a mercury (Hg) exposure gradient. The aim of this study was to assess the effects of Hg and methylmercury (MeHg) on the general physiological condition of fish as well as to gauge the relationship between MeHg and the glutathione (GSH) system in metal-contaminated and reference sites using a series of biomarkers. Walleye from Lake Malartic had the highest liver MeHg concentrations, exhibited lower hepatosomatic indices (HSI) and lower glutathione S-transferase (GST) activity. HSI was negatively related to liver total Hg concentrations in walleye (R2=0.33, n=108, P<0.0001). Glutathione reductase (GR) and GST activity for walleye from Lake Malartic were related to HSI (R2=0.38, n=25, P=0.0010; R2=0.46, n=27, P<0.0001, respectively). In Lake Desjardins-East, where perch had the highest liver MeHg concentrations, glutathione peroxidase selenium dependent activity (GSH-Px SD) and GST activity were negatively related to liver MeHg concentrations (R2=0.39, n=21, P=0.0026; R2=0.22, n=21, P=0.0298, respectively). This study suggests that Hg may induce adverse effects on the physiology and cellular metabolism of walleye and perch at environmentally relevant concentrations.     

Development of a biomarker-based index for assessing the ecotoxic potential of aquatic sites.

The use of biochemical or physiological measurements as indicators of ecotoxicity is under constant development and has the advantage of delineating effects before the appearance of disease. However, these biomarkers are often part of a battery of tests, and it is difficult to integrate them together to gain an overall view of an organism's health. The aim of this study was to develop an index that could integrate the data derived from a battery of biomarkers for application to both spatial and temporal studies. Mya arenaria clams were collected at different sites along the Saguenay Fjord (Quebec, Canada). Six biomarkers were measured: metallothioneins, DNA strand breakage, lipid peroxidation, vitellin-like proteins, phagocytosis, and non-specific esterase activity in haemocytes. A biomarker index was obtained by summing the biomarker values expressed in term of classes. Classes were determined by a distribution-free approach derived from the theory of rough sets. The results of the spatial study show that the index values discriminated well between contaminated and uncontaminated sites. The highly polluted sites had the highest index values (18 compared with a reference value of 14). In the temporal study, the index was also able to highlight possible contamination-induced alterations, even though the interpretation of temporal variation is complicated by natural variations occurring throughout the year. A control chart approach is proposed for determining contaminated sites in both spatial and temporal surveys.     

Chemosymbiotic bivalves from the mud volcanoes of the Gulf of Cadiz, NE Atlantic, with descriptions of new species of Solemyidae, Lucinidae and Vesicomyidae

The chemosymbiotic bivalves collected from the mud volcanoes of the Gulf of Cadiz are reviewed. Of the thirteen species closely associated with chemosynthetic settings two Solemyidae, Solemya (Petrasma) elarraichensissp. n. and Acharax gadiraesp. n., one Lucinidae, Lucinoma asapheussp. n., and one Vesicomyidae, Isorropodon megadesmussp. n. are described and compared to close relatives of their respective families. The biodiversity and distribution of the chemosymbiotic bivalves in the Gulf of Cadiz are discussed and compared to the available information from other cold seeps in the Eastern Atlantic and Mediterranean. Although there is considerable similarity at the genus level between seep/mud volcano fields in the Eastern Atlantic and Mediterranean, there is little overlap at the species level. This indicates a high degree of endemism within chemosymbiotic bivalve assemblages.     

Lipid‐based indicators of nutritional condition in juvenile sole Solea solea

Effects of the feeding level on juvenile Solea solea (Linnaeus, 1758) lipid‐related indicators of nutritional condition were assessed in a 25‐day trial experiment. Morphometric indices (K and HSI), liver lipid content and fatty acids (FA) clusters were good indicators of the feeding level, while individual FA showed highly dynamic variation patterns. Additionally, liver indices were more sensitive to feeding stress than were muscle measurements. HSI determination (and K) is fairly straightforward, with lower costs and simple procedures. Nevertheless, liver lipid content is also an important indicator of the feeding level and constitutes a direct measurement of food utilization and energy storage capacity. The individual variability and complex dynamics of FA with the feeding level are disadvantageous for use in FA analysis even for early‐warning signals of feed limitation. Overall, S. solea nutritional condition can be suitably assessed by morphometric indices and lipid content, especially considering liver measurements (HSI and liver lipid content), provided that the fish development stage is accounted for (i.e. gonad development and reproductive cycle).     

A proteome resource of ovarian cancer ascites: integrated proteomic and bioinformatic analyses to identify putative biomarkers

Epithelial ovarian cancer is the most lethal gynecological malignancy, and disease-specific biomarkers are urgently needed to improve diagnosis, prognosis, and to predict and monitor treatment efficiency. We present an in-depth proteomic analysis of selected biochemical fractions of human ovarian cancer ascites, resulting in the stringent and confident identification of over 2500 proteins. Rigorous filter schemes were applied to objectively minimize the number of false-positive identifications, and we only report proteins with substantial peptide evidence. Integrated computational analysis of the ascites proteome combined with several recently published proteomic data sets of human plasma, urine, 59 ovarian cancer related microarray data sets, and protein-protein interactions from the Interologous Interaction Database I (2)D ( http://ophid.utoronto.ca/i2d) resulted in a short-list of 80 putative biomarkers. The presented proteomics analysis provides a significant resource for ovarian cancer research, and a framework for biomarker discovery.     

Field studies on the relation between the accumulation of heavy metals and metabolic and HSR in the bearded horse mussel Modiolus barbatus

The present study aims to examine whether the seasonal changes in the levels of heavy metals Cd and Pb in the gills of sublittoral mussel Modiolus barbatus from populations distributed in Thermaikos Gulf are correlated to seasonal molecular (Heat Shock Response) and metabolic stress responses of this species. Our results indicate a season effect in the accumulation of heavy metals in the gills of bivalves in the area of Thermaikos Gulf, possibly implicating biological (reproductive cycle), natural (rivers' flow) and anthropogenic factors. Among the members of heat shock proteins (Hsps) examined, inducible Hsp70 seemed to play a major cytoprotective role against toxicity of metals. However, bivalves' tolerance against heavy metals may decrease during warming, arising further questions for their survival in context of global warming.     

Multi response biomarker approach in the crab Carcinus aestuarii experimentally exposed to benzo a pyrene, polychlorobiphenyls and methyl mercury

The aim of this study was to test a multi response biomarker approach for evaluating toxicological risk due to some of the main contaminants in the Mediterranean benzo a pyrene, polychlorobiphenyls and methyl mercury , using the Mediterranean crab Carcinus aestuarii as bioindicator organism. Forty crabs were injected with different doses of these contaminants. Several molecular, biochemical and genotoxic biomarkers were tested in different tissues and biological materials. The main conclusions were: 1 hepatopancreas, gills, haemolymph and excreta seem to be useful for biomarker studies in this species; 2 several biochemical, molecular and genotoxic biomarkers were found suitable for testing in these tissues; 3 several biomarkers were found suitable for evaluating chemical stress due to different Mediterranean contaminants.