Immunotoxicity of the xenoestrogen 4-nonylphenol to the cockle Cerastoderma glaucum

The in vivo effects of 4-nonylphenol (NP) on functional responses of haemocytes from the cockle Cerastoderma glaucum were investigated after 7 days exposure to sublethal NP concentrations (0, 0+acetone, 0.0125, 0.025, 0.05 and 0.1 mg/l NP). Haemocytes from both controls and exposed cockles were collected, and the effects of NP on total haemocyte count (THC) and volume of circulating cells, intracellular superoxide anion (O(2)(-)) levels, acid phosphatase and lysozyme-like activities in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. Exposure of cockles to 0.1mg/l NP significantly increased THC (p<0.05) with respect to controls. Analysis of haemocyte size frequency distribution showed that the haemocyte fraction of about 7-8 microm in diameter and 250 femtolitres in volume increased markedly in cockles exposed to the highest NP concentration tested. Apoptosis resulting in cell volume reduction in NP-exposed animals cannot be excluded. No statistically significant variation in intracellular O(2)(-) levels was observed. Conversely, significant increases (p<0.05) in acid phosphatase activity were observed in CFH from 0.05 and 0.1mg/l NP-exposed animals; no significant differences in enzyme activity were recorded in HL. Lysozyme-like activity also increased significantly in CFH from cockles exposed to 0.05 mg/l NP (p<0.05) and 0.1 mg/l NP (p<0.001). Instead, lysozyme-like activity decreased significantly (p<0.05) in the HL of animals exposed to 0.05 mg/lNP. Our results suggest that NP induces variations in the functional responses of haemocytes of C. glaucum, mainly by reducing cell membrane stability and promoting cell degranulation.     

Lethal and estrogenic effects of 4-nonylphenol in the cockle Cerastoderma glaucum

The lethal and sublethal effects of the xenoestrogen 4-nonylphenol (NP) were evaluated in the cockle Cerastoderma glaucum. In a 96-h lethality test, bivalves were exposed to 0, 0+ acetone, 0.19, 0.38, 0.75, 1.5 and 3.0 mg NP/l. The 96-h LC(50) value was 0.3mg NP/l. No mortality was observed at 0.1 mg NP/l. The potential estrogenicity of NP was studied in both sexually undifferentiated (resting phase) and differentiated (pre-spawning phase) cockles, exposed for 7 and 14 days to 0, 0+ acetone, 0.0125, 0.025, 0.05, and 0.1 mg NP/l. Vitellogenin (Vg)-like protein levels were determined in both haemolymph and digestive gland by the alkali-labile phosphate (ALP) assay. In the resting phase, exposure for 7 days to 0.1 mg NP/l resulted in significant increases in ALP in both haemolymph and digestive gland, compared with controls. A significant increase was also observed in digestive gland of animals exposed to 0.0125 mg NP/l-exposed animals. After 14 days of exposure, haemolymph ALP levels were significantly increased in exposed animals at all NP concentrations tested, whereas no difference was recorded in digestive gland. In the pre-spawning phase, exposure for 7 days to NP significantly increased ALP levels in haemolymph from males exposed at all NP concentrations tested, whereas no significant variations were found in haemolymph from females. NP (0.05 and 0.1 mg/l) was also shown to increase ALP concentrations significantly in digestive gland of males, but not in those of females. Likewise, after 14 days' exposure, ALP levels significantly increased in haemolymph from males only at 0.1 mg NP/l. Conversely, NP caused significant increases in ALP levels in digestive gland from both males (at all NP concentrations tested) and females (at 0.025 and 0.1 mg NP/l). These results demonstrate that NP induces Vg synthesis in C. glaucum. Interestingly, males were more responsive to NP than females.     

Lethal and estrogenic effects of 4-nonylphenol in the cockle Cerastoderma glaucum

The lethal and sublethal effects of the xenoestrogen 4-nonylphenol (NP) were evaluated in the cockle Cerastoderma glaucum. In a 96-h lethality test, bivalves were exposed to 0, 0+ acetone, 0.19, 0.38, 0.75, 1.5 and 3.0 mg NP/l. The 96-h LC₅₀ value was 0.3 mg NP/l. No mortality was observed at 0.1 mg NP/l. The potential estrogenicity of NP was studied in both sexually undifferentiated (resting phase) and differentiated (pre-spawning phase) cockles, exposed for 7 and 14 days to 0, 0+ acetone, 0.0125, 0.025, 0.05, and 0.1 mg NP/l. Vitellogenin (Vg)-like protein levels were determined in both haemolymph and digestive gland by the alkali-labile phosphate (ALP) assay. In the resting phase, exposure for 7 days to 0.1 mg NP/l resulted in significant increases in ALP in both haemolymph and digestive gland, compared with controls. A significant increase was also observed in digestive gland of animals exposed to 0.0125 mg NP/l-exposed animals. After 14 days of exposure, haemolymph ALP levels were significantly increased in exposed animals at all NP concentrations tested, whereas no difference was recorded in digestive gland. In the pre-spawning phase, exposure for 7 days to NP significantly increased ALP levels in haemolymph from males exposed at all NP concentrations tested, whereas no significant variations were found in haemolymph from females. NP (0.05 and 0.1 mg/l) was also shown to increase ALP concentrations significantly in digestive gland of males, but not in those of females. Likewise, after 14 days’ exposure, ALP levels significantly increased in haemolymph from males only at 0.1 mg NP/l. Conversely, NP caused significant increases in ALP levels in digestive gland from both males (at all NP concentrations tested) and females (at 0.025 and 0.1 mg NP/l). These results demonstrate that NP induces Vg synthesis in C. glaucum. Interestingly, males were more responsive to NP than females.     

Lethal and estrogenic effects of 4-nonylphenol in the cockle Cerastoderma glaucum

The lethal and sublethal effects of the xenoestrogen 4-nonylphenol (NP) were evaluated in the cockle Cerastoderma glaucum. In a 96-h lethality test, bivalves were exposed to 0, 0+ acetone, 0.19, 0.38, 0.75, 1.5 and 3.0 mg NP/l. The 96-h LC₅₀ value was 0.3 mg NP/l. No mortality was observed at 0.1 mg NP/l. The potential estrogenicity of NP was studied in both sexually undifferentiated (resting phase) and differentiated (pre-spawning phase) cockles, exposed for 7 and 14 days to 0, 0+ acetone, 0.0125, 0.025, 0.05, and 0.1 mg NP/l. Vitellogenin (Vg)-like protein levels were determined in both haemolymph and digestive gland by the alkali-labile phosphate (ALP) assay. In the resting phase, exposure for 7 days to 0.1 mg NP/l resulted in significant increases in ALP in both haemolymph and digestive gland, compared with controls. A significant increase was also observed in digestive gland of animals exposed to 0.0125 mg NP/l-exposed animals. After 14 days of exposure, haemolymph ALP levels were significantly increased in exposed animals at all NP concentrations tested, whereas no difference was recorded in digestive gland. In the pre-spawning phase, exposure for 7 days to NP significantly increased ALP levels in haemolymph from males exposed at all NP concentrations tested, whereas no significant variations were found in haemolymph from females. NP (0.05 and 0.1 mg/l) was also shown to increase ALP concentrations significantly in digestive gland of males, but not in those of females. Likewise, after 14 days’ exposure, ALP levels significantly increased in haemolymph from males only at 0.1 mg NP/l. Conversely, NP caused significant increases in ALP levels in digestive gland from both males (at all NP concentrations tested) and females (at 0.025 and 0.1 mg NP/l). These results demonstrate that NP induces Vg synthesis in C. glaucum. Interestingly, males were more responsive to NP than females.     

Incorporation of in situ exposure and biomarkers response in clams Ruditapes philippinarum for assessment of metal pollution in coastal areas from the Maluan Bay of China

The clams Ruditapes philippinarum were used to assess the impact of metal contaminants when transplanted to seven study sites along the Maluan Bay (China). Metal concentrations in digestive gland tissues of clams after 7-day in situ cage exposure were determined in conjunction with antioxidant enzyme activities. The results showed the importance of specific antioxidant biomarkers to assess complex pollutant mixtures and their good correlations to the pollutant compositions of deployment sites. Multivariate analysis indicated causal relationship between the chemicals at each study site and the biochemical "response" of the caged clams at these sites and demonstrated the potential presence of two different contaminant sources. This study suggested that the incorporation of tissue residue analysis with biomarkers response in caged clams together with factor analysis can be a useful biomonitoring tool for the identification of causal toxic pollutants and the assessment of complex metal pollutions in marine coastal environment.     

Effects of 4-nonylphenol exposure in mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii) with particular emphasis on vitellogenin induction

Since it is often difficult to estimate possible adverse effects due to contamination in selected ecosystems, multi-species biomonitoring may provide more information, taking into account different routes of exposure, ecological roles and metabolic capabilities of animals. In this context, we exposed for 7 days the mussel Mytilus galloprovincialis and the crab Carcinus aestuarii to 4-nonylphenol (NP), a well-known xenoestrogen. In mussels (0-0.2 mg NP l(-1)), we measured NP bioaccumulation in soft tissues and vitellogenin (Vg)-like protein levels in digestive glands from both males and females by the alkali-labile phosphate assay (ALP). As no reference data were available for crab exposure, the NP 96-h LC(50) value was previously determined. Then, in sublethally exposed (0-1.0 mg NP l(-1)) male crabs, NP bioaccumulation and Vg levels were measured in hemolymph, gonads and digestive gland. Bioaccumulation of NP increased from 43 to 371 microg g(-1) d.w. in mussels, and from 3.6 to 37 microg g(-1) d.w. in crabs, depending on the NP concentration in water. Dose-dependent Vg-like protein induction was observed in both species, appearing to be related to NP bioaccumulation, although a partial decrease was recorded at the highest concentration tested. A similar trend was observed in both digestive gland and gonad of exposed crabs; Vg increased to a lesser extent, although significantly, in hemolymph. Results demonstrated that NP induces Vg synthesis both in male and female mussels, as well as in male crabs. On the basis of the responsiveness of both species investigated, a multi-species approach is indicated in biomonitoring programmes.     

Immunotoxicity of the xenoestrogen 4-nonylphenol to the cockle Cerastoderma glaucum

The in vivo effects of 4-nonylphenol (NP) on functional responses of haemocytes from the cockle Cerastoderma glaucum were investigated after 7 days exposure to sublethal NP concentrations (0, 0+acetone, 0.0125, 0.025, 0.05 and 0.1 mg/l NP). Haemocytes from both controls and exposed cockles were collected, and the effects of NP on total haemocyte count (THC) and volume of circulating cells, intracellular superoxide anion () levels, acid phosphatase and lysozyme-like activities in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. Exposure of cockles to 0.1 mg/l NP significantly increased THC (p < 0.05) with respect to controls. Analysis of haemocyte size frequency distribution showed that the haemocyte fraction of about 7–8 μm in diameter and 250 femtolitres in volume increased markedly in cockles exposed to the highest NP concentration tested. Apoptosis resulting in cell volume reduction in NP-exposed animals cannot be excluded. No statistically significant variation in intracellular levels was observed. Conversely, significant increases (p < 0.05) in acid phosphatase activity were observed in CFH from 0.05 and 0.1 mg/l NP-exposed animals; no significant differences in enzyme activity were recorded in HL. Lysozyme-like activity also increased significantly in CFH from cockles exposed to 0.05 mg/l NP (p < 0.05) and 0.1 mg/l NP (p < 0.001). Instead, lysozyme-like activity decreased significantly (p < 0.05) in the HL of animals exposed to 0.05 mg/l NP. Our results suggest that NP induces variations in the functional responses of haemocytes of C. glaucum, mainly by reducing cell membrane stability and promoting cell degranulation.     

Variation of Antioxidant System in Pinus armandii under Elevated O3 in an Entire Growth Season

Impact of elevated O₃ (80 nmol mol^?1) was studied on antioxidant system in the previous‐year needles (P‐needles) of Pinus armandii under the background of global climate change. CAT (catalase) did not play an important role in defensing O₃ pollution, as no significant change was observed in its activity in P‐needles exposed to elevated O₃ during the entire growth season. Ascorbate (AsA) was more sensitive to elevated O₃, as the O₃‐induced decrease in its content was observed at Day 45. It suggests that AsA consumption was a crucial way for protecting against O₃ pollution. After 75 days of O₃ exposure, increase in superoxide dismutase and ascorbate peroxidase activities occurred in P‐needles (p < 0.05), whereas glutathione reductase activity was significantly increased after 105 days of O₃ treatment (p < 0.05). Dehydroascorbate reductase and monoascorbate reductase activities also increased after 75 days of elevated O₃ exposure. However, the elevated antioxidant capability did not suppress the generation of reactive oxygen species, and as a result, increase in malondialdehyde content was noted (p < 0.05). Noticeably, no change was observed in electrolyte leakage during the entire growth season under elevated O₃ fumigation. It indicates that O₃‐induced oxidative stress was not severe in the needles of P. armandii. In other words, P. armandii has certain tolerance to O₃ pollution.     

Effects of 4-nonylphenol exposure in mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii) with particular emphasis on vitellogenin induction

Since it is often difficult to estimate possible adverse effects due to contamination in selected ecosystems, multi-species biomonitoring may provide more information, taking into account different routes of exposure, ecological roles and metabolic capabilities of animals. In this context, we exposed for 7 days the mussel Mytilus galloprovincialis and the crab Carcinus aestuarii to 4-nonylphenol (NP), a well-known xenoestrogen. In mussels (0–0.2 mg NP l⁻¹), we measured NP bioaccumulation in soft tissues and vitellogenin (Vg)-like protein levels in digestive glands from both males and females by the alkali-labile phosphate assay (ALP). As no reference data were available for crab exposure, the NP 96-h LC₅₀ value was previously determined. Then, in sublethally exposed (0–1.0 mg NP l⁻¹) male crabs, NP bioaccumulation and Vg levels were measured in hemolymph, gonads and digestive gland. Bioaccumulation of NP increased from 43 to 371 μg g⁻¹ d.w. in mussels, and from 3.6 to 37 μg g⁻¹ d.w. in crabs, depending on the NP concentration in water. Dose-dependent Vg-like protein induction was observed in both species, appearing to be related to NP bioaccumulation, although a partial decrease was recorded at the highest concentration tested. A similar trend was observed in both digestive gland and gonad of exposed crabs; Vg increased to a lesser extent, although significantly, in hemolymph. Results demonstrated that NP induces Vg synthesis both in male and female mussels, as well as in male crabs. On the basis of the responsiveness of both species investigated, a multi-species approach is indicated in biomonitoring programmes.     

In vitro effects of nonylphenol on functional responses of haemocytes of the colonial ascidian Botryllus schlosseri

The effects of nonylphenol (NP) on functional responses of haemocytes of the colonial ascidian Botryllus schlosseri were evaluated. Haemocytes were exposed to 0.1, 1 and 10 μM NP and the effects on haemocyte viability, adhesion, morphology, lysosomal membrane stability, phagocytic activity and early apoptosis were evaluated. Haemocyte viability and adhesion were not affected by NP. Phagocytic activity and the amoebocytic index decreased significantly at all the concentrations tested, while exposure of haemocytes to 1 and 10 μM NP caused a significant increase in the diffusion of Neutral Red into the cytosol. The percentage of cells positive to Annexin-V (indicative of early apoptosis) increased significantly at 1 and 10 μM NP. Results obtained suggested a relationship between NP and alterations in functional responses of haemocytes in B. schlosseri. Biomarkers measured resulted sensitive, rapid and reproducible, even if their responsiveness will be evaluated after in vivo exposure of animals to NP.     

Effects of 4-nonylphenol exposure in mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii) with particular emphasis on vitellogenin induction

Since it is often difficult to estimate possible adverse effects due to contamination in selected ecosystems, multi-species biomonitoring may provide more information, taking into account different routes of exposure, ecological roles and metabolic capabilities of animals. In this context, we exposed for 7 days the mussel Mytilus galloprovincialis and the crab Carcinus aestuarii to 4-nonylphenol (NP), a well-known xenoestrogen. In mussels (0–0.2 mg NP l⁻¹), we measured NP bioaccumulation in soft tissues and vitellogenin (Vg)-like protein levels in digestive glands from both males and females by the alkali-labile phosphate assay (ALP). As no reference data were available for crab exposure, the NP 96-h LC₅₀ value was previously determined. Then, in sublethally exposed (0–1.0 mg NP l⁻¹) male crabs, NP bioaccumulation and Vg levels were measured in hemolymph, gonads and digestive gland. Bioaccumulation of NP increased from 43 to 371 μg g⁻¹ d.w. in mussels, and from 3.6 to 37 μg g⁻¹ d.w. in crabs, depending on the NP concentration in water. Dose-dependent Vg-like protein induction was observed in both species, appearing to be related to NP bioaccumulation, although a partial decrease was recorded at the highest concentration tested. A similar trend was observed in both digestive gland and gonad of exposed crabs; Vg increased to a lesser extent, although significantly, in hemolymph. Results demonstrated that NP induces Vg synthesis both in male and female mussels, as well as in male crabs. On the basis of the responsiveness of both species investigated, a multi-species approach is indicated in biomonitoring programmes.     

Variation of antioxidant enzyme activities of sprat (Sprattus sprattus) larvae and organic contaminant levels in mixed zooplankton from the southern North Sea.

Sprat (Sprattus sprattus) larvae and mixed zooplankton were collected from sites in the southern North Sea across three transects running north, north-west and west from the Elbe and Weser estuaries (Germany). Sprat larval sub-cellular fractions (13,500 g supernatants) were assayed for the antixoxidant enzymes catalase (EC 1.11.1.6) and superoxide dismutase (SOD; EC 1.15.1.1), and levels of polychlorobiphenyls (PCBs), p,p'-DDE and polycyclic aromatic hydrocarbons (PAHs) were determined in the mixed zooplankton (fish larval diet). Higher levels (p < 0.05) of SOD and catalase activities were observed at sites closest to estuaries corresponding to sites with the highest levels (p < 0.05) of total PCBs and p,p'-DDE. Antioxidant enzymes activities decreased in samples collected from sites further from the estuaries across a northern and north-western transect; however this was not observed across a western transect. Larval antioxidant enzyme activities are discussed in relation to potential processes affecting them including plankton contaminant level distributions.     

Concentrations of unsubstituted polycyclic aromatic hydrocarbons in softshell clams from Coos Bay,Oregon, USA

Concentrations of benzo[a]pyrene (BAP) were measured in subpopulations of softshell clams, Mya arenaria, from four intertidal sites in Coos Bay from June 1976 to June 1978. Subsequently, concentrations of 15 unsubstituted polynuclear aromatic hydrocarbons (PNAH) were determined in two subpopulations from September 1978 to August 1979. There were significant differences between BAP concentrations in clams from the four sites. For the two-year period, they were highest in clams inhabiting areas adjacent to the industrialized bayfront and lowest in clams inhabiting more remote areas. There were no significant seasonal variations in BAP concentrations during this period. During the 1978–1979 study, the average total PNAH concentration in clams from the bayfront area was 555.1 μg kg^?1 compared to 76.3 μg kg^?1 for clams from a more remote environment. In general, PNAH concentrations were lowest in the fall-winter and highest during the spring-summer.     

Relationships between tissue concentrations of chlorinated hydrocarbons (polychlorinated biphenyls and chlorinated pesticides) and antioxidative responses of marine mussels, Perna viridis

Marine mussels, Perna viridis, were transplanted from a reference site to various polluted sites around Hong Kong. After 30 d of exposure, antioxidative responses in the gills and hepatopancreas and tissue concentrations of chlorinated hydrocarbons [polychlorinated biphenyls (PCBs) and chlorinated pesticides (CPs)] were determined for individual mussels. Glutathione S transferase (GST) and glutathione (GSH) were positively correlated with tissue PCB concentrations. Only one of the enzymatic antioxidants, glutathione peroxidase (GPx), showed significant response to tissue PCB. No significant correlation was found between tissue concentrations of chlorinated hydrocarbons and other enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and NADPH DT-diaphorase (DT-d). Oxidative stress, measured as thiobarbituric acid reactive substances, was correlated with chlorinated pesticide concentrations in tissues. This study demonstrated a correlation between GST/GSH and chlorinated hydrocarbons. The apparent lack of correlation between trace organic pollutants and some of the enzymatic antioxidants may be due to the inhibitory effects caused by these chemicals. The above results suggest that more investigations are needed before these enzymes can be used as biomarkers.     

Acetylcholinesterase as a biomarker of exposure to neurotoxic compounds in the clam Tapes philippinarum from the Lagoon of Venice

Measurement of acetylcholinesterase (AChE) is widely used as a biomarker of exposure to neurotoxic compounds in aquatic environments. In the present study, AChE activity was monitored in the clam Tapes philippinarum in the Lagoon of Venice, with the aim of evaluating the possible exposure of animals to neurotoxic compounds. Two sampling strategies were followed. In the first case, temporal and spatial variations of AChE activity were measured in clams collected seasonally (from October 2003 to June 2004) in 3 sites of the Lagoon: Campalto (site 1), near a sewage treatment plant, Marghera (site 2), a highly polluted area, and Poveglia (site 3), a reference site. The condition index (CI) of clams was also calculated. In the second case, AChE activity was measured in clams collected at the end of seasonal samplings in 12 sites located throughout the Lagoon: areas licensed for clam farming (sites 5 to 11), nearby canals characterised by waste waters from agricultural areas (sites 4, 12, 13, 14), and a reference site outside the Lagoon (site 15). Results revealed a significant reduction in AChE activity in clams from Marghera in October, January and April, indicating their probable exposure to neurotoxic compounds originating in the industrial zone of Porto Marghera. A seasonal trend in AChE activity was also observed, enzyme activity being higher in January and lower in June. Clams from Campalto generally had higher CI values than those from Marghera and Poveglia. However, no particular correlation between AChE activity and CI was found. Lastly, marked differences in AChE activity were recorded when comparing enzyme activity of clams from various sites in the Lagoon of Venice with those of clams collected at site 15. Significant reductions in AChE activity were observed in animals collected in both nearby canals and licensed areas, indicating the homogeneous spatial distribution of potentially neurotoxic compounds throughout the Lagoon. It is probable that sediment mobilisation caused by clam harvesting and the passage of shipping plays an important role in resuspension of persistent contaminants having neurotoxic activity.     

Toxic contaminants and their biological effects in coastal waters of Xiamen,China. II. Biomarkers and embryo malformation rates as indicators of pollution stress in fish

Baseline information is presented on embryo malformation rate and biomarkers in fish as indicators of sub-lethal stress caused by pollution in coastal waters of Xiamen, PR China. Fish and eggs were sampled from several areas in Xiamen coastal waters (Xiamen Harbour, Maluan and Tongan Bays and East Channel), where varying levels of pollutant input have been documented. Comparative sampling was done at a "cleaner" reference site at Dongshan Island. Embryonic malformation rates, which indicate general water quality, varied with location and species of fish, and exceeded background levels for unpolluted waters (assumed approximately 5%) by up to eightfold at some sites. Generally, sites around Xiamen Harbour show signs of poor water quality having highest mean levels of embryo deformity (20-30%) and these decreased towards open waters (Tongan Bay, Eastern Channel) where abnormalities approached background levels. An indication that toxic contaminants may be having a localised effect in the region, particularly in the harbour was reinforced by the biomarker assays. However, activities of the biomarkers ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase in fish livers indicate no clear pattern, and there is no evidence that fish from the four sampling areas have been more or less exposed to PAHs and other compounds that induce these biomarkers. Antioxidant biomarkers (glutathione peroxidase, catalase, superoxide dismutase, and reduced glutathione) suggest that exposure to xenobiotics appears to be lowest in Dongshan and Maluan and highest in the harbour and Tongan. Inhibition of acetylcholinesterase in fish muscle indicated possible effects by organophosphate and carbamate pesticides in Xiamen waters and these effects may be greatest in the area of the harbour.     

Sublethal effect of naphthalene on lipid peroxidation and antioxidant status in the edible marine crab Scylla serrata

The effect of naphthalene on lipid peroxidation and antioxidants status was studied in hepatopancreas, haemolymph and ovary of the Scylla serrata with reference to the active vitellogenic stage. There was an overall increase in lipid peroxidation activity in the tested samples. In contrast, the enzymatic (catalase, glutathione peroxidase, superoxide dismutase) and non-enzymatic antioxidants (vitamins C, E and glutathione) showed decreased activities for hepatopancreas, haemolymph and ovary.     

Relationships between tissue concentrations of chlorinated hydrocarbons (polychlorinated biphenyls and chlorinated pesticides) and antioxidative responses of marine mussels,Perna viridis

Marine mussels, Perna viridis, were transplanted from a reference site to various polluted sites around Hong Kong. After 30 d of exposure, antioxidative responses in the gills and hepatopancreas and tissue concentrations of chlorinated hydrocarbons [polychlorinated biphenyls (PCBs) and chlorinated pesticides (CPs)] were determined for individual mussels. Glutathione S transferase (GST) and glutathione (GSH) were positively correlated with tissue PCB concentrations. Only one of the enzymatic antioxidants, glutathione peroxidase (GPx), showed significant response to tissue PCB. No significant correlation was found between tissue concentrations of chlorinated hydrocarbons and other enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and NADPH DT-diaphorase (DT-d). Oxidative stress, measured as thiobarbituric acid reactive substances, was correlated with chlorinated pesticide concentrations in tissues. This study demonstrated a correlation between GST/GSH and chlorinated hydrocarbons. The apparent lack of correlation between trace organic pollutants and some of the enzymatic antioxidants may be due to the inhibitory effects caused by these chemicals. The above results suggest that more investigations are needed before these enzymes can be used as biomarkers.     

Application of oxidative stress indices in natural populations of Perna viridis as biomarker of environmental pollution

Oxidative stress indices were measured in gills and digestive glands of Perna viridis collected from three coastal locations in Goa i.e., Bambolim, Marmugao Harbour and Malim. In addition to lipid peroxidation, the activities of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase and two non-enzymatic antioxidants (ascorbic acid and reduced glutathione) were investigated in order to understand their variation with respect to pollution status of the sampling locations. We observed a significant increase in lipid peroxidation and antioxidant enzymes of both the tissues at Mormugao Harbour and Malim, suggesting that the animals at these two locations are at higher level of oxidative stress as compared to those at Bambolim. Conversely, low levels of non-enzymatic antioxidants such as ascorbic acid and reduced glutathione, observed at Mormugao Harbour and Malim indicate that the animals may use these compounds to counteract stress in the tissues. This study shows that changes in lipid peroxidation, superoxide dismutase, catalase, glutathione reductase, glutathione S-transferase and reduced glutathione in tissues of P. viridis can be used as molecular biomarkers in environmental monitoring programs.