Assessment of ecosystem health disturbance in mangrove-lined Caribbean coastal systems using the oyster Crassostrea rhizophorae as sentinel species.

This investigation was aimed at contributing to develop a suitable multi-biomarker approach for pollution monitoring in mangrove-lined Caribbean coastal systems using as sentinel species, the mangrove cupped oyster, Crassostrea rhizophorae. A pilot field study was carried out in 8 localities (3 in Nicaragua; 5 in Colombia), characterized by different environmental conditions and subjected to different levels and types of pollution. Samples were collected in the rainy and dry seasons of 2012-2013. The biological effects at different levels of biological complexity (Stress-on-Stress response, reproduction, condition index, tissue-level biomarkers and histopathology) were determined as indicators of health disturbance, integrated as IBR/n index, and compared with tissue burdens of contaminants in order to achieve an integrative biomonitoring approach. Though modulated by natural variables and confounding factors, different indicators of oyster health, alone and in combination, were related to the presence of different profiles and levels of contaminants present at low-to-moderate levels. Different mixtures of persistent (As, Cd, PAHs) and emerging chemical pollutants (musk fragrances), in combination with different levels of organic and particulate matter resulting from seasonal oceanographic variability and sewage discharges, and environmental factors (salinity, temperature) elicited a different degree of disturbance in ecosystem health condition, as reflected in sentinel C. rhizophorae. As a result, IBR/n was correlated with pollution indices, even though the levels of biological indicators of health disturbance and pollutants were low-to-moderate, and seasonality and the incidence of confounding factors were remarkable. Our study supports the use of simple methodological approaches to diagnose anomalies in the health status of oysters from different localities and to identify potential causing agents and reflect disturbances in ecosystem health. Consequently, the easy methodological approach used herein is useful for the assessment of health disturbance in a variety of mangrove-lined Caribbean coastal systems using mangrove cupped oysters as sentinel species.

Recent findings on phenoloxidases in bivalves.

The production of melanin is a complex process involving biochemical cascades, such as the pro-phenoloxidase (proPO) system, and enzymes, such as phenoloxidases (POs). Different studies have shown a strong correlation between the decrease in PO activities and the occurrence of diseases in bivalve invertebrates, leading to mortalities in the host. Results of these studies suggest that POs could play a fundamental role in defense mechanisms in bivalves. This article reviews the fundamental knowledge on the proPO system in bivalves and the methods used to assess PO activities. Finally, this is the first report on the major findings of laboratory and field studies that indicate that a type of PO in bivalves, the laccase enzyme, is inducible and involved in the 1) immune 2) antioxidant and 3) detoxification roles in bivalves, and might be an ecological potential biomarker of environmental stress.

Persistent organic pollutants in a marine bivalve on the Marennes-Oléron Bay and the Gironde Estuary (French Atlantic Coast) - part 2: potential biological effects.

Contaminant effects on defence responses of ecologically and economically important organisms, such as the Pacific oyster Crassostrea gigas, are likely to influence their ability to resist infectious diseases, particularly at the young stages. The aim of this study was to explore the potential relationships between organic contaminants accumulated in the soft tissues of juvenile oysters, defence responses and physiological condition. Oysters were transplanted during summer and winter periods in different sites in the Marennes-Oléron Bay, the first area of oyster production in France, and in the Gironde Estuary, the biggest estuary in Occidental Europe. Amongst the battery of biochemical and physiological biomarkers applied in the present work [superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), malondyaldehyde (MDA), catecholase, laccase and lysozyme in gills, digestive glands, mantle and haemolymph, glycogen, proteins and lipids in the digestive gland and the condition index at the whole-organism level], MDA and lysozyme in the digestive gland and SOD, GPx and laccase in plasma contributed in order to significantly discriminate the sites in which oysters bioaccumulated different levels of heavy polycyclic aromatic hydrocarbons (HPAHs), polychlorobiphenyls (PCBs), polybromodiphenylethers (PBDEs), dichlorodiphenyltrichloroethanes (DDTs) and lindane. These results strengthen the hypothesis that it is possible to differentiate sites depending on their contamination levels and biological effects by carrying out studies with transplanted juvenile oysters. In addition, correlations were found between antioxidant and immune-defence responses, and PAH and DDT body burdens in the first area of oyster production in France (the Marennes-Oléron Bay) and where considerable oyster mortalities have been reported. This result suggests that the presence of organic chemical contaminants in the Marennes-Oléron Bay may influence defence responses in juveniles of C. gigas, and, therefore, could influence their ability to resist infectious diseases.

Persistent organic pollutants in a marine bivalve on the Marennes-Oléron Bay and the Gironde Estuary (French Atlantic Coast) - part 1: bioaccumulation.

The aim of this study was to determine 1) the relevance of using the Pacific oyster Crassostrea gigas as a sentinel organism, at a juvenile stage, for polycyclic aromatic hydrocarbon (PAH) and persistent organic pollutant (polychlorobiphenyl, PCB, polybromodimethylether, PBDE, and organochlorine pesticide, OCP) contamination, 2) the potential levels of chemical organic contamination in the Marennes-Oléron Bay, and their potential sources and 3) the potential influence of physiological or environmental factors on contaminant body burdens in oysters. To this end, juvenile oysters purchased from an oyster hatchery were transplanted to a reference site, in Bouin, and to different transplantation sites in the Marennes-Oléron Bay, the first oyster production area in France, and in the Gironde Estuary, the biggest estuary in Occidental Europe. Transplantations were done during summer and winter. Whole oyster soft tissues from each site were analysed for PAHs, PCBs, PBDEs and OCPs. Results obtained with a transplantation period of 3months suggest that C. gigas, at the juvenile stage, is a relevant sentinel organism for short-term assessment of contamination for these contaminants. In addition, no significant effects of physiological factors on contaminant body burdens were observed. Principal component analysis revealed two distinct groups of contaminants (PAHs and OPCs, and PCBs and PBDEs) and three groups of sites: 1) the reference site, 2) Les Palles (LP) and Boyard (BOY) in winter and 3) all the other sites. The group of LP and BOY was clearly defined by the levels of PAHs and OCPs, suggesting higher levels of contamination of these chemical compounds on these sites, potentially due to local contamination sources. In addition, no relevant effects of physiological or environmental factors on contaminant body burdens were observed. Results suggest also a predominance of contaminants related to agricultural activities along the Marennes-Oléron Bay, and therefore, further studies on the presence of pesticides in this region should be considered.

Detection of early effects of a single herbicide (diuron) and a mix of herbicides and pharmaceuticals (diuron, isoproturon, ibuprofen) on immunological parameters of Pacific oyster (Crassostrea gigas) spat.

In the context of massive summer mortality events of the Pacific oyster Crassostrea gigas, the aim of this study was to investigate the early effects on genes, enzymes and haemocyte parameters implicated in immune defence mechanisms in C. gigas oysters exposed to a potentially hostile environment, i.e. to an herbicide alone or within a mixture. Following 2 h of exposure to the herbicide diuron at 1 µg L(-1), the repression of different genes implicated in immune defence mechanisms in the haemocytes and the inhibition of enzyme activities, such as laccase-type phenoloxidase (PO) in the plasma, were observed. The inhibition of superoxide dismutase (SOD) activity in the plasma was also observed after 6 and 24 h of exposure. In the mixture with the herbicides diuron and isoproturon, and the pharmaceutical ibuprofen, catecholase-type PO activity in the plasma and the percentage of phagocytosis in the haemocytes were reduced after 6 h of exposure. Our results showed that early effects on molecular, biochemical and cellular parameters can be detected in the presence of diuron alone or within a mixture, giving an insight of its potential effect in situations that can be found in natural environments, i.e. relatively high concentrations for short periods of time.

Bioaccumulation and metabolisation of (14)C-pyrene by the Pacific oyster Crassostrea gigas exposed via seawater.

The first objective of this study was to determine the bioaccumulation kinetics of pyrene in the soft tissues of Crassostrea gigas (mantle, muscle, gills, digestive gland, and the remaining soft tissues). As bivalves can biotransform hydrocarbons in more polar compounds (metabolites) that are more easily excreted, the second objective was to investigate the oyster capacity to metabolize pyrene into its metabolite, the 1-hydroxypyrene. To these ends, oysters were exposed 24 h to waterborne (14)C-pyrene then placed in depuration conditions for 15 d. Oysters efficiently bioaccumulated pyrene in their soft tissues and equilibrium was reached within the exposure time. The metabolite1-hydroxypyrene was also detected in oyster tissues but represented only 4-14% of the parent pyrene. At the end of the exposure period, the gills and the mantle showed the highest pyrene proportion of total soft tissue content, i.e. 47% and 26%, respectively. After 15 d of depuration, the mantle contained 32% and 30% of the remaining pyrene and 1-hydroxypyrene, respectively. As C. gigas did not display a high capacity for metabolizing pyrene, it can be considered as a good bioindicator species to survey and monitor pyrene contamination in the coastal marine environment.

Enhanced immunological and detoxification responses in Pacific oysters, Crassostrea gigas, exposed to chemically dispersed oil.

The aim of this study was to evaluate the effects of chemically dispersed oil on an economically and ecologically important species inhabiting coasts and estuaries, the Pacific oyster Crassostrea gigas. Studies were carried out with juveniles, known to generally be more sensitive to environmental stress than adults. A set of enzyme activities involved in immune defence mechanisms and detoxification processes, i.e. superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), catecholase-type phenoloxidase (PO), laccase-type PO and lysozyme were analysed in different oyster tissues, i.e. the gills, digestive gland and mantle, and in the plasma and the haemoycte lysate supernatant (HLS) of the haemolymph. Results indicated that total PAH body burdens were 2.7 times higher in the presence than in the absence of the chemical dispersant. After 2 days of exposure to chemically dispersed oil, alkylated naphthalenes accounted for 55% of the total PAH body burden, whereas alkylated fluorenes and alkylated dibenzothiophenes accounted for 80% when the chemical dispersant was absent. Importantly, a higher number of enzyme activities were modified when oil was chemically dispersed, especially in the plasma and gills. Moreover, independently of the presence or absence of chemical dispersant, oil exposure generally inhibited enzyme activities in the gills and plasma, while they were generally activated in the mantle and haemocytes. These results suggest that the gills and plasma constitute sensitive compartments in C. gigas, and that the mantle and haemocytes may play an important role in protection against xenobiotics. Among the six enzyme activities that were analysed in these body compartments, five were modulated in the chemical dispersion (CD) treatment while only half of the enzyme activities were modulated in the mechanical dispersion treatment. Furthermore, CD treatment effects were often observed following exposure, but also during depuration periods. These results suggest that immune and/or detoxification responses are likely to be affected when dispersants are used to treat oil spills in shallow waters.

Phenoloxidase activation in the embryo of the common cuttlefish Sepia officinalis and responses to the Ag and Cu exposure.

The prophenoloxidase (proPO) system catalyzing the melanin production is considered as implicated in the innate immune system in invertebrates. The phenoloxidase (PO)-like activity was detected in the cuttlefish embryo sampled at the end of the organogenesis and few hours before hatching. Various modulators of the PO activity were used to assess the triggering of the proPO activating system. The results demonstrated the evidence of a true PO activity in the cuttlefish embryo. However, SDS and LPS granted contrasting effects on the PO-like activity between the developmental stages suggesting a progressive maturation of the proPO system from the embryonic to the juvenile stages. In eggs exposed to dissolved trace metals all along the embryonic development, Ag (1.2 microg L(-1)) inhibited the PO-like activity in the cuttlefish embryo except at hatching time, suggesting the synthesis of a new "juvenile" form of the PO enzyme. In similar conditions as for Ag, Cu (230 microg L(-1)) stimulated and then inhibited the PO-like activity according to a progressive metal accumulation within the egg and suggesting the occurrence of a threshold, above which the toxicity of the essential metal reduced the PO activity.