Effect of acute cadmium exposure on oxidative stress and antioxidant system of the scallop Aequipecten tehuelchus.

This study aimed to assess the impact of acute exposure (96 h) to Cd in gills, digestive gland and muscle of the Tehuelche scallop Aequipecten tehuelchus from San José gulf in Patagonia, Argentina. Scallops were exposed to Cd concentrations of 0, 25, 50, 100, 150, 204, 275, 371, and 500 µg/L, and mortality rates were recorded after 96 h of exposure. Surviving organisms were analyzed for the biochemical response through reactive oxygen and nitrogen species (RONS), activities of catalase (CAT) and glutathione-S-transferase (GST), metallothioneins (MT), lipid peroxidation (LPO) and liposoluble antioxidants α-tocopherol (α-T) and ß-carotene (ß-C). The mean lethal concentration (LC50) was 155.8 µg Cd/L, a lower value than other scallops' species, showing that A. tehuelchus has a particular sensitivity to Cd. In the three tissues, at all exposure concentrations, there was no significant response in RONS levels, GST activity or LPO. Nevertheless, CAT activity and α-T levels decreased in the gills but increased in the digestive gland, with no significant response in the muscle. Two-way ANOVA revealed a significant interaction between Cd concentration and tissue on MT, which increased significantly in gills, decreased in digestive gland with 100 compared to 50 µg Cd/L; whereas in muscle a significant increase was observed with 25 µg Cd/L compared to control. The results show a significant effect of Cd in scallop's gills on CAT activity and α-T levels, highlighting this tissue as the primary target against relevant concentrations of metal in seawater. The effect on digestive gland and muscle was minimal. The overall results suggest that Cd toxicity is tissue-specific. This study will help reduce the existence knowledge gap regarding potential impacts of acute exposure to Cd in a bivalve species with high ecological and commercial importance, as well as identifying the most responsive biomarkers associated with Cd stress for monitoring assessment.

Biochemical response and tissue-specific accumulation of scallop Aequipecten tehuelchus from Patagonia, Argentina after exposure to inorganic arsenic.

This study investigates the effects of different inorganic arsenic (As III) concentrations (0, 125, 500 and 1000 µg As/L) following two exposure times (7 and 14 days) on gills, digestive gland and muscle of scallop Aequipecten tehuelchus from Patagonia, Argentina. A biochemical approach was used to investigate oxidative stress-related parameters after different As concentrations and exposure times. Although the accumulation of As was of the same order of magnitude in all tissues, the results showed distinct tissue-specific oxidative responses to this metalloid. Furthermore, the variation in exposure time had no significant effect on As accumulation in any of the three tissues. In gills, despite no reactive oxygen and nitrogen species (RONS) were detected, there was an increase in catalase (CAT) activity and metallothionein (MT) levels. Conversely, digestive gland showed RONS production without a rise in CAT and glutathione S-transferases (GST) activities, but with an increase in MT levels. In muscle, RONS production and CAT activity kept constant or decreased, while MT levels remained unchanged. In addition, exposure time demonstrated its critical role in gills by influencing the response of CAT, GST and MT, particularly at high As concentrations, while exposure time did not affect the biochemical stress parameters in the digestive gland and muscle. Interestingly, neither concentration of As produced lipid damage, showing the effectiveness of the antioxidant mechanisms to avoid it. These results emphasize that A. tehuelchus exhibited no time-dependent effects in response to As exposure, while showing tissue-specific responses characterized by significant concentration-dependent effects of As. This study provides a comprehensive insight by considering the combined effects of time and concentration of a contaminant and distinguishing its effects on specific tissues, a dimension often overlooked in the existing literature. Subsequent studies should prioritize the analysis of additional contaminants in species with increased sensitivity.

Evidences of metabolic alterations and cellular damage in different tissues of scallops Aequipecten tehuelchus exposed to cadmium.

Scallops Aequipecten tehuelchus (Patagonia, Argentina) were exposed to 0, 2, 5 and 12 µg Cd/L for 7 and 14 days, causing in digestive gland a significant production of reactive oxygen and nitrogen species (RONS), induction of catalase (CAT) and glutathione S-transferase (GST) activities and metallothioneins (MT) synthesis. In gills, there was inhibition of GST and induction of CAT, MT and α-tocopherol (α-Toc). In muscle, a significant increment of MT was also registered and inhibition of CAT. Lipid peroxidation, measured as TBARS, was not promoted in any tissue. More significant effects were observed in digestive gland than in gills and muscle, evidencing the critical role of digestive gland in Cd accumulation and metabolisation. This research would evidence dose-dependent effects of Cd on MT, GST, CAT and α-Toc in the three organs assayed, as well as a time-dependent effect of Cd on the response of CAT, GST and TBARS in digestive gland.

The Chubut River estuary as a source of microplastics and other anthropogenic particles into the Southwestern Atlantic Ocean.

This is the first report of anthropogenic particles (APs), including microplastics and synthetic, semi-synthetic and anthropogenically-altered natural fibers, in water and sediment of the Chubut River estuary. This river is the main source of freshwater in Chubut Province (Patagonia, Argentina), where wastes and pollutants are poured and finally end in the Atlantic Ocean. The average concentration in surface and bottom water samples was 5.5 items/L, while in sediment was 175.4 items/kg dw. Raman's analysis identified particles dominated by polyethylene terephthalate (PET) (35.5 %), dye signature only (18.5) and anthropogenic cellulose (10 %). Fibers were the prevalent shape (83 %), and the chemical identification evidenced a textile origin. The highest APs concentration was found in sediments from the site with the finest grain size and the greatest amount of organic matter. Present results will provide a baseline for future studies and raise public and governmental awareness.

Chitosan performance during Paralytic Shellfish Toxins (PST) depuration of Mytilus chilensis exposed to Alexandrium catenella.

Blooms of the dinoflagellate Alexandrium catenella, which produces Paralytic Shellfish Toxins (PST), generate serious socio-economic consequences for mariculture in Chile, especially for the production of Mytilus chilensis and other bivalves. Palliative strategies, such as the depuration of mussels in enriched water with chitosan offer encouraging prospects against the advance of contaminated areas and toxin persistence. Adult mussels were fed with A. catenella for 20 days and then were allowed to depurate using chitosan as facilitator, for the same period. Intoxicated mussels showed a reduction in feeding activity and rapid PST accumulation in 20 days (C = 451.5t + 1,673.6, R2 = 0.55 p = 0.008). Not enough evidence was found to indicate a positive effect of chitosan in mussel depuration after 20 days (C = -311.1t + 8,462.4, R2 = 0.8 p = 0.001). At the end of the study, toxicity was higher than 800 µg STX eq kg-1. C2 and GTX4 analogues were the most abundant in the dinoflagellate strain, while C2 and C1 were the most accumulated in mussels. The presence of C1 was notorious during depuration, as the persistence of GTX2,3. GTX5 was only detected in A. catenella, while STX was only present in mussels. Mussel sensitivity to the presence of the toxic dinoflagellate was observed in the present study. The biotransformation, selective elimination and epimerization processes were deduced from intoxication and depuration experiments.

Multiple-Biomarker Approach in a Commercial Marine Scallop from San Jose gulf (Patagonia, Argentina) for Health Status Assessment.

The health status of the commercial Tehuelche scallop Aequipecten tehuelchus from San Román and El Riacho in San José gulf (Patagonia, Argentina) was evaluated through biomarkers widely used in ecotoxicological applications. Natural levels of arsenic (As) and cadmium (Cd) were measured to determine their potential relationships with fluctuations of several oxidative stress biomarkers in the scallop. Oxidative biomarkers, such as catalase (CAT), glutathione S-transferase (GST), superoxide dismutase (SOD), metallothioneins (MT), reactive oxygen species (ROS), α-tocopherol (α-T), and lipid peroxidation (LPO) through thiobarbituric acid reactive substances (TBARS) and lipid radical (LR∙), were measured in gills, digestive gland, and muscle of Tehuelche scallop in winter (August 2015) and summer (January 2016). Levels of As and Cd and of most of the biomarkers (SOD, ROS, TBARS, and LR∙) showed strong seasonal variability in the three tissues. In general, the highest values were recorded in digestive gland. The Integrated Biomarker Response index indicated that the most stressed condition of A. tehuelchus was in summer in San Román. Additionally, the Integrated Biomarker Response index showed a strong relationship among tissues and As and Cd accumulation. This kind of approach could be used as an integrated tool to identify the health status of scallop A. tehuelchus from San José gulf.

Potential mobility assessment of metals in salt marsh sediments from San Antonio Bay.

The BCR method was applied on sediments from the salt marsh of San Antonio Bay (SAB). It presents several channels among which the Encerrado is the most important and is impacted by abandoned mining wastes. The pseudototal concentrations of metals measured within this channel were relatively higher than in outer sites, and according to the Igeo index, its contamination level was low. The metal distribution in the different phases of sediment particles showed that the residual component, considered the safest from the environmental point of view, accounted for most of the Fe, Cd, Cu, and Zn contents. Conversely, Pb was mainly in the non-residual component as part of the reducible fraction, thus constituting the main environmental hazard among the studied elements. The predominance of residual and reducible fractions indicated a historic contamination of metal such as Pb, Cu, and Zn from the mining wastes. The low exchangeable and oxidizable fractions would indicate no actual input of metals.

Metals from mine waste as potential cause of oxidative stress in burrowing crab Neohelice granulata from San Antonio bay.

The Natural Protected Area San Antonio bay is of particular importance for its congregation of migratory shorebirds and it has been declared one of the Western Hemisphere Shorebird Reserve Network International site (WHSRN). Present study represents the first assessment of variation on oxidative stress biomarkers in male crab Neohelice granulata from San Antonio bay (Río Negro, Argentina) under field conditions, associated mainly to metal contamination coming from passive mining wastes. Three sites were sampled once every three months from November 2012 to August 2013 within this sea inlet (Pile, Fishery and Port) and a control site at the southeast of the bay (Punta Perdices). Accumulation of Ni, Zn, Cr and Al varied only with seasons although without a constant trend, meanwhile Cd, Cu and Pb also varied among sites being highest in Pile and Port. Biochemical results indicated that variations in catalase activity was only site specific being maximum in Pile; meanwhile lipid radical, α-tocopherol and metallothioneins were only seasonal specific being higher in autumn and winter. Seasonal variation was also found for total thioles, being the content higher in summer and autumn than in winter. Correlation analysis revealed that malondialdehyde and α-tocopherol have a positive association with Al and negative with Ni, meanwhile GST has a positive association with Fe. Crabs from the closest area to the waste pile did not exhibit a differentiated oxidative pressure despite the higher accumulation of metals. It is possible that crabs from contaminated areas have developed a tolerance to metals, indicating a strong ecotoxicological selective pressure. More studies are needed to assess whether there is a transfer of metals through the food chain.

Biomarkers of environmental stress in gills of ribbed mussel Aulacomya atra atra (Nuevo Gulf, Northern Patagonia).

In this study, we assessed in gills of native ribbed mussels Aulacomya atra atra from three sites within Nuevo Gulf (Northern Patagonia) several biomarkers such as reactive oxygen species (ROS), lipid radicals (LR), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST) and metallothionein (MT). Furthermore, concentrations of main trace metals (Fe, Al, Zn, Cu, Cd and Pb) were quantified in mussel tissue. Results showed significant induction of SOD, GST, MT and MDA, as well as, higher concentration of Fe, Al and Cd in winter than in summer. The high MDA content measured in mussels from Folías Wreck seemed to be caused by the very high levels of Fe that would come from the corrosion of the vessel. Mussels from the control site Punta Cuevas presented the lowest levels of Cd and the highest of Al in winter. Despite positive correlations were found between Al and GST and MT, no spatial differentiation was detected in those biomarkers. On the other hand, MT was only related to Al been most likely influenced by environmental variables than by the trace metals. It has to be highlighted that the relationship detected among water temperature, nutrients and antioxidant responses in gills is probably related to the fact that this tissue is in direct contact with water and it is sensitive to its fluctuations. Taking into account that mussel gill is a tissue actively proliferating and the first target of contaminants present in water, so that changes in its antioxidant system can provide an earlier warning signal than in other tissues.

Assessment of antioxidant responses and trace metal accumulation by digestive gland of ribbed mussel Aulacomya atra atra from Northern Patagonia.

Seasonal and spatial variability of trace metal concentrations and of a battery of antioxidant parameters were evaluated in digestive gland of the ribbed mussel Aulacomya atra atra. Fe, Al and Cu accumulated in tissue exhibited maximum values in winter, coinciding partially with the highest labile concentrations of Fe and Cu in sediment. Metals, as other pollutants, are known to influence the oxidative status of organisms and antioxidant enzymes have been often proposed as biomarkers of contaminant effects. Seasonal variations of trace metals did not appear to influence those of biochemical parameters, which generally showed an opposite trend with higher enzymatic activities in summer when trace metal concentrations were lower. Organisms from Punta Cuevas (control site) showed higher induction of reactive oxygen species production than those from both considered impacted sites, suggesting the possibility of some biochemical adaptation in organisms or a higher modulation of environmental and physiological factors on antioxidant responses than levels of trace metals. This study, which is the first in the area in this matter, showed that seasonal variations of potential biomarkers should be incorporated into interpretation of long-term biomonitoring studies in this marine coastal ecosystem.

Trace metal content in sediments and autochthonous intertidal organisms from two adjacent bays near Ushuaia, Beagle Channel (Argentina).

The aim of this work was to monitor levels of Cd, Cu, Pb, Zn and Fe in sediments, mussels (Mytilus edulis chilensis) and limpets (Nacella magellanica) from the Industrial zone (IZ); fuel dock (FD) and Ushuaia Peninsula (UP) on the Beagle Channel. In sediments, seasonal variations showed high values of Cu and Pb in spring and Zn in autumn. Comparing among sites, Cd concentration was superior in UP (2.07 µg/g); while Pb was maximum in FD (41.00 µg/g). In mussels, a higher bioaccumulation in winter was found. Mussels from UP showed the highest bioaccumulation of Cu (5.95 µg/g) and those from FD presented the highest of Zn (170.15 µg/g). A seasonal trend was not found for limpets, while differences among sites were observed for Cd being the highest at IZ (3.02 µg/g). Although pollution level found was low, anthropic activities at the studied sites could result in deterioration, further monitoring is recommended.

Heavy metal concentrations and biomarkers of oxidative stress in native mussels (Mytilus edulis chilensis) from Beagle Channel coast (Tierra del Fuego, Argentina).

The aim of this study was to evaluate the usefulness of oxidative stress biomarkers of pollution in native mussels Mytilus edulis chilensis from the Beagle Channel. Spatial and seasonal variations of catalase, glutathione-S-transferase and lipid peroxidation in gills and digestive gland were analyzed in relation to environmental parameters, heavy metals in sediment and in tissue. Four sites with anthropogenic impact and a control site were selected and monitored during the four seasons of 2007. We found significant differences among sites in concentrations of dissolved nutrients and heavy metals in sediments, with the highest values recorded at sites with anthropogenic pressure. Different patterns were observed between concentrations of metals in tissues and in sediments suggesting differences in bioavailability. There were also significant differences in biomarker responses among sites, despite the strong seasonal variability. Our results showed relatively moderate levels of pollution in the study area as a result of urban influences.

Seasonal and pollution-induced variations in biomarkers of transplanted mussels within the Beagle Channel.

The occurrence of biomarker variations linked to environmental factors makes it difficult to distinguish the effect of pollution. In an attempt to evaluate spatial and seasonal effects of environmental parameters on biomarker responses, mussels Mytilus edulis chilensis coming from an aquaculture farm were transplanted to several points within Ushuaia Bay (Beagle Channel) for 6 weeks in summer and winter. Activities of superoxide dismutase, catalase, glutathione-S-transferase and levels of lipid peroxidation were measured in gills and digestive gland. Cu, Zn, Fe, Cd and Pb concentrations were also assessed. Results indicated a significant effect of seasons on biological responses as well as in metal bioaccumulation showing the influence of natural factors such as dissolved oxygen, temperature and food availability. The interdependence of those environmental factors is important for the homeostasis of thermoconformers, especially regarding their oxidative metabolism and should also be taken into consideration to distinguish natural from pollution-induced variations.

Heavy metals monitoring in the southernmost mussel farm of the world (Beagle Channel, Argentina).

Water quality surrounding the mussel farm of Mytilus edulis chilensis at Brown Bay (Beagle Channel) was evaluated. The levels of five heavy metals in sediment and in gill and digestive gland of mussels were examined to consider potential risks to human health. Cd showed the highest enrichment factor in relation to its level in Earth crust (3.85-21.58), which could be related to an upwelling phenomenon. A seasonal trend was found regarding metal bioaccumulation, being higher in winter than in summer. The bioaccumulation pattern in gill was Zn>Fe>Cu>Cd, meanwhile in digestive gland was Fe>Zn>Cu>Cd. Despite Pb was measured in sediment (15.59-23.91 microg/g dw), it was not available for being incorporated by mussels. In all cases it was below the detection limit (2.37 microg/g dw). With regard to human consumption of mussels from Brown Bay, none of the elements analyzed should cause concern for consumers. Values measured in tissue mussels were below the limit of 10 microg/g dw for Cd and Pb established by SENASA for molluscs. Considering that studied mussels are for human consumption and the relatively high levels of metals in sediment may vary their availability if physical parameters changes, periodical monitoring must be carried out to avoid human risks and to produce food in a responsible manner that complies with the food safety standards.

Biomarkers and heavy metal bioaccumulation in mussels transplanted to coastal waters of the Beagle Channel.

Mussels coming from a mussel farm at Brown Bay (Beagle Channel) were transplanted to four sites inside Ushuaia Bay for 2 and 4 weeks. The objective of this study was to assess the quality of coastal waters of Ushuaia Bay by measuring catalase activity, lipid peroxidation, total lipid content, bioaccumulation of heavy metals and condition index in transplanted mussel Mytilus edulis chilensis. Biomarkers except condition index showed significant differences among exposure times as well as among tissues. Digestive gland presented the highest catalase activity, malondialdehyde level and total lipid content. Digestive gland also was the main target tissue of accumulation of iron and copper, while gill accumulated the highest levels of zinc. A principal component analyzes with the whole set of data allowed to separate stations based on physicochemical conditions and biochemical responses of each studied area.

Heavy metal toxicity in Exosphaeroma gigas (Crustacea, Isopoda) from the coastal zone of Beagle Channel.

Acute toxicity of copper, cadmium, and zinc on isopod Exosphaeroma gigas was evaluated at 20 per thousand and 30 per thousand salinity. Six concentrations were assayed to estimate effective concentration of the toxicant that affects 50% of tested animal (EC50), while physiological responses and bioaccumulation were determined at 0.42 and 1.95 mg L(-1) of each metal. The following toxicity orders were obtained: Cd>or=Cu>Zn at 20 per thousand salinity and Cu>Zn>or=Cd at 30 per thousand salinity. Copper treatments showed a reduction in oxygen consumption at 30 per thousand salinity, while an opposite trend was observed at 20 per thousand salinity. Zinc caused dissimilar effects, while in cadmium significant reduction was only registered in 0.42(20 per thousand) mg L(-1). Ammonia excretion was generally higher in treatments than control at 20 per thousand salinity. At 30 per thousand salinity, excretion did not change in relation to control or diminished. In general terms, O:N atomic ratios indicated a preponderant protein metabolism. Bioaccumulation of assayed metals was higher at lowest salinity and increased with increasing toxic concentrations.