First evidence of altered vitellogenin-like protein levels in clam Tapes philippinarum and in cockle Cerastoderma glaucum from the Lagoon of Venice

Possible xenoestrogenic effects were investigated, for the first time, in two bivalve species from the Lagoon of Venice (Italy): the clam Tapes philippinarum and the cockle Cerastoderma glaucum. Bivalves were collected far from their reproductive phase at the very early stage of gametogenesis (January), and in the pre-spawning period (June) in six sites. Vitellogenin (Vg)-like proteins (a biomarker of exposure to estrogenic compounds) were measured by the alkali-labile phosphate method (ALP), in both haemolymph and digestive gland from males and females (when it was possible to distinguish sex by microscopic observation of gonadal tissue). Haemolymph calcium concentrations (a parameter considered closely related to the presence of Vg-like proteins) and the bivalve condition index (CI) were also measured. In both seasons, bivalves collected at Campalto (near a sewage treatment plant) and Marghera (a highly contaminated area) had higher Vg-like protein levels, particularly in haemolymph, than animals from the other sampling sites. Interestingly, CI had high values in these polluted sites. In June only, Vg-like proteins and Ca(2+) levels in haemolymph exhibited similar trends in both bivalve species at most sampling sites. The responsiveness of bivalves to environmental xenoestrogens was higher in June, allowing better discrimination among sites. The present study demonstrates that animals from highly polluted areas have increased Vg-like protein levels. As endocrine disruption due to exposure to estrogenic compounds may cause fertility reduction, alterations in the sex ratio, and a decrease in reproductive rate, a condition of potential risk for bivalve populations in estuarine areas is highlighted.     

First evidence of altered vitellogenin-like protein levels in clam Tapes philippinarum and in cockle Cerastoderma glaucum from the Lagoon of Venice

Possible xenoestrogenic effects were investigated, for the first time, in two bivalve species from the Lagoon of Venice (Italy): the clam Tapes philippinarum and the cockle Cerastoderma glaucum. Bivalves were collected far from their reproductive phase at the very early stage of gametogenesis (January), and in the pre-spawning period (June) in six sites. Vitellogenin (Vg)-like proteins (a biomarker of exposure to estrogenic compounds) were measured by the alkali-labile phosphate method (ALP), in both haemolymph and digestive gland from males and females (when it was possible to distinguish sex by microscopic observation of gonadal tissue). Haemolymph calcium concentrations (a parameter considered closely related to the presence of Vg-like proteins) and the bivalve condition index (CI) were also measured. In both seasons, bivalves collected at Campalto (near a sewage treatment plant) and Marghera (a highly contaminated area) had higher Vg-like protein levels, particularly in haemolymph, than animals from the other sampling sites. Interestingly, CI had high values in these polluted sites. In June only, Vg-like proteins and Ca²⁺ levels in haemolymph exhibited similar trends in both bivalve species at most sampling sites. The responsiveness of bivalves to environmental xenoestrogens was higher in June, allowing better discrimination among sites. The present study demonstrates that animals from highly polluted areas have increased Vg-like protein levels. As endocrine disruption due to exposure to estrogenic compounds may cause fertility reduction, alterations in the sex ratio, and a decrease in reproductive rate, a condition of potential risk for bivalve populations in estuarine areas is highlighted.     

First evidence of altered vitellogenin-like protein levels in clam Tapes philippinarum and in cockle Cerastoderma glaucum from the Lagoon of Venice

Possible xenoestrogenic effects were investigated, for the first time, in two bivalve species from the Lagoon of Venice (Italy): the clam Tapes philippinarum and the cockle Cerastoderma glaucum. Bivalves were collected far from their reproductive phase at the very early stage of gametogenesis (January), and in the pre-spawning period (June) in six sites. Vitellogenin (Vg)-like proteins (a biomarker of exposure to estrogenic compounds) were measured by the alkali-labile phosphate method (ALP), in both haemolymph and digestive gland from males and females (when it was possible to distinguish sex by microscopic observation of gonadal tissue). Haemolymph calcium concentrations (a parameter considered closely related to the presence of Vg-like proteins) and the bivalve condition index (CI) were also measured. In both seasons, bivalves collected at Campalto (near a sewage treatment plant) and Marghera (a highly contaminated area) had higher Vg-like protein levels, particularly in haemolymph, than animals from the other sampling sites. Interestingly, CI had high values in these polluted sites. In June only, Vg-like proteins and Ca²⁺ levels in haemolymph exhibited similar trends in both bivalve species at most sampling sites. The responsiveness of bivalves to environmental xenoestrogens was higher in June, allowing better discrimination among sites. The present study demonstrates that animals from highly polluted areas have increased Vg-like protein levels. As endocrine disruption due to exposure to estrogenic compounds may cause fertility reduction, alterations in the sex ratio, and a decrease in reproductive rate, a condition of potential risk for bivalve populations in estuarine areas is highlighted.     

Polycyclic aromatic hydrocarbons (PAHs) in transplanted Manila clams (Tapes philippinarum) from the Lagoon of Venice as assessed by PAHs/shell weight index: A preliminary study

Variation of polycyclic aromatic hydrocarbons (PAHs) levels was assessed in Tapes philippinarum from the Lagoon of Venice. Clams were transplanted from a polluted area next to Porto Marghera to two rearing areas of the Southern Lagoon. Analyses of PAHs were made in sediments and clams by GC/MS at first sampling and after 30, 60 and 180 days. Principal component analysis was performed to elucidate bioaccumulation and depuration pattern and input sources. Biota-Sediment-Accumulation-Factor (BSAF) was applied to evaluate the PAHs input sources from sediment. Condition index was calculated to compare the seasonal variation of clam tissue to PAHs levels. To propose results not affected by seasonal changes in flesh weight of clams, the approach based on the calculation of PAHs/SW index was applied. From the results, it was concluded that PAHs/SW index is more recommendable to asses temporal variation of PAHs levels in Manila clams.     

Polycyclic aromatic hydrocarbons (PAHs) in transplanted Manila clams (Tapes philippinarum) from the Lagoon of Venice as assessed by PAHs/shell weight index: a preliminary study

Variation of polycyclic aromatic hydrocarbons (PAHs) levels was assessed in Tapes philippinarum from the Lagoon of Venice. Clams were transplanted from a polluted area next to Porto Marghera to two rearing areas of the Southern Lagoon. Analyses of PAHs were made in sediments and clams by GC/MS at first sampling and after 30, 60 and 180 days. Principal component analysis was performed to elucidate bioaccumulation and depuration pattern and input sources. Biota-Sediment-Accumulation-Factor (BSAF) was applied to evaluate the PAHs input sources from sediment. Condition index was calculated to compare the seasonal variation of clam tissue to PAHs levels. To propose results not affected by seasonal changes in flesh weight of clams, the approach based on the calculation of PAHs/SW index was applied. From the results, it was concluded that PAHs/SW index is more recommendable to asses temporal variation of PAHs levels in Manila clams.     

Polycyclic aromatic hydrocarbons (PAHs) in transplanted Manila clams (Tapes philippinarum) from the Lagoon of Venice as assessed by PAHs/shell weight index: A preliminary study

Variation of polycyclic aromatic hydrocarbons (PAHs) levels was assessed in Tapes philippinarum from the Lagoon of Venice. Clams were transplanted from a polluted area next to Porto Marghera to two rearing areas of the Southern Lagoon. Analyses of PAHs were made in sediments and clams by GC/MS at first sampling and after 30, 60 and 180 days. Principal component analysis was performed to elucidate bioaccumulation and depuration pattern and input sources. Biota-Sediment-Accumulation-Factor (BSAF) was applied to evaluate the PAHs input sources from sediment. Condition index was calculated to compare the seasonal variation of clam tissue to PAHs levels. To propose results not affected by seasonal changes in flesh weight of clams, the approach based on the calculation of PAHs/SW index was applied. From the results, it was concluded that PAHs/SW index is more recommendable to asses temporal variation of PAHs levels in Manila clams.     

Sediment chemical contamination of a shallow water area close to the industrial zone of Porto Marghera (Venice Lagoon, Italy)

The industrial zone of Porto Marghera is one the most important "contaminated sites of national interest" (SIN) in Italy, being identified as an area of high environmental risk. The site includes a wide shallow water area of the Venice Lagoon extending toward the city of Venice, which was investigated in order to acquire information on the pollutant level and distribution. Grain-size, heavy metal, nutrient, and organic micropollutant concentrations were determined in the surface sediment layers (0-5 and 5-10 cm depths) of 51 sites. A generally low contamination was found, except for Hg concentration, which increases the toxicological risk in most of the sites of the area, according to the results of a comparison with Sediment Quality Guidelines. A heavy pollution fingerprint (Cd, Cu, Hg, Pb, and Zn up to 15.2, 257, 11.9, 248, and 3010 mg/kg d.w., respectively) was instead found near the Tresse Island, which is ascribed to the spill of pollutants from the contaminated sediment disposed therein. Grain-size and heavy metal profiles down to a depth of 40 cm in eight selected sites, finally show a probable decrease of the pollution affecting the area in recent years.     

Sediment chemical contamination of a shallow water area close to the industrial zone of Porto Marghera (Venice Lagoon,Italy)

The industrial zone of Porto Marghera is one the most important “contaminated sites of national interest” (SIN) in Italy, being identified as an area of high environmental risk. The site includes a wide shallow water area of the Venice Lagoon extending toward the city of Venice, which was investigated in order to acquire information on the pollutant level and distribution. Grain-size, heavy metal, nutrient, and organic micropollutant concentrations were determined in the surface sediment layers (0–5 and 5–10 cm depths) of 51 sites. A generally low contamination was found, except for Hg concentration, which increases the toxicological risk in most of the sites of the area, according to the results of a comparison with Sediment Quality Guidelines. A heavy pollution fingerprint (Cd, Cu, Hg, Pb, and Zn up to 15.2, 257, 11.9, 248, and 3010 mg/kg d.w., respectively) was instead found near the Tresse Island, which is ascribed to the spill of pollutants from the contaminated sediment disposed therein. Grain-size and heavy metal profiles down to a depth of 40 cm in eight selected sites, finally show a probable decrease of the pollution affecting the area in recent years.     

Antioxidant responses to variations in dissolved oxygen of Scapharca inaequivalvis and Tapes philippinarum, two bivalve species from the lagoon of Venice

The aim of the present work was to determine the activities of selected antioxidant enzymes (SOD, Se-GPX, CAT) in two species of bivalves, Scapharca inaequivalvis and Tapes philippinarum, from two sites of the lagoon of Venice that are characterized by different pO(2) (Marghera and Chioggia). The specimens were collected at four times during a 1-year period. In the two species studied, enzyme activities were found to be present in both digestive glands and gills, but with some species-specific differences that may also represent a different adaptation to seasonal variations. The presence of high SOD activities in the gills of both species may be related to their physiological role in respiration. Scapharca inaequivalvis is less sensitive than T. philippinarum to environmental changes, perhaps due to the presence of hemoglobins in this species. Moreover, in the digestive gland of T. philippinarum we found a significant negative correlation between the activities of SOD and GPX that may indicate the presence of oxidative stress. Some correlations between temperature/dissolved oxygen and antioxidant enzyme activity were present in specimens sampled in Marghera. Only GPX adequately responded to changes in dissolved oxygen and temperature, while the decrease in the activity of SOD and CAT in winter may be directly responsible for an enhanced susceptibility of mussels to oxidative stress during this period. We can conclude that the observed differences between Chioggia and Marghera are due to different concentrations of dissolved oxygen. Marghera is an appropriate location to study seasonal variations in water temperature. In fact, in this site, the differences between hot and cold months are quite evident.     

Use of the clam Asaphis deflorata as a potential indicator of organochlorine bioaccumulation in Hong Kong coastal sediments

In order to test its ability as a bioaccumulator, the deposit feeding clam Asaphis deflorata, collected from a clean coastal area in Hong Kong (Ding Chau, Ma On Shan), was exposed to field-collected sediments representing a gradient of contamination. Sediments were collected from four sites, namely Shek O in the southeast of Hong Kong Island, Ap Lei Chau in Aberdeen district, Tai Kok Tsui in the Mong Kok waterfront, and To Kwa Wan near Kowloon City pier. Shek O was the cleanest and To Kwa Wan was the most polluted site. Replicate 30L fiberglass tanks containing equal numbers of organisms were maintained for 28 days, with equal quantities of sediment and filtered seawater. Prior to this the animals were depurated for 10 days in clean sediment. During the 28-day holding period, sampling of both sediment and clams was performed on days 0, 10 and 28, along with condition index measurements. Concentrations of organochlorine pesticides and PCBs in sediments showed that Shek O was the cleanest of the sites, followed by Ap Lei Chau, Tai Kok Tsui and To Kwa Wan. There were marked differences among the sites, especially in PAH concentrations, with To Kwa Wan showing extremely high sediment concentrations. Following exposure, the biota concentrations of various organochlorine pesticides and PCBs suggested that there was sediment concentration-dependent accumulation by A. deflorata on day 10 of the experiment. However, there was no significant increase on day 28 for any of the compounds tested. Contaminant concentrations in clams feeding on Shek O sediments were minimal and almost the same on all sampling days, indicating the presence of only baseline contaminant concentrations. The condition indices (CI) of the clams on various sampling days showed that the animals were relatively more stable in sediment from Shek O. There were some deaths in all the test tanks (including those containing Shek O sediments). In addition, clams appeared to bury faster in the cleaner Shek O sediment and remained buried, unlike those in the sediment from the other locations, where some remained predominantly on the surface. Nevertheless, despite not completely burying, the clams in other test sediments still exhibited sediment concentration-dependent tissue accumulation. The results of the investigation indicate the possibility of developing a standard test protocol for bioaccumulation by sediment dwelling invertebrates, for practical use in Hong Kong, using A. deflorata.     

Determination of polychlorobiphenyl congeners (PCBs) in the surface water of the Venice lagoon

The purpose of this study is to evaluate the degree of PCB contamination of Venice lagoon water. The PCB determination was made on the filtrate ("dissolved PCBs") and on the particulate collected on the filters ("particulate PCBs") by continuous liquid-liquid extraction and sonication, respectively, and by HRGC-LRMS analysis. An estimation of the procedural and "working" blanks was also made. Water samples were collected at six sites, selected to represent all the pollution situations present in the lagoon, and at a site in the Adriatic Sea. The range of the total sum ("dissolved plus particulate") of 49 congener concentrations falls between 355 and 1868 pgl(-1); the "dissolved" fraction concentrations (250-792 pgl(-1)) are greater at six sites than that of the "particulate" fraction (105-1273 pgl(-1)). Chemometric analyses suggest that the PCB congener pattern in the Venice Lagoon is different from that in the Adriatic Sea.     

Accumulation of polychlorinated dibenzo-p-dioxins and dibenzofurans in sediments of the Venice Lagoon and the industrial area of Porto Marghera

Two sediment cores collected from the central Venice Lagoon and five from the canals of the nearby industrial area were studied in order to assess the temporal evolution of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) delivery and present trends. The most polluted site is C12, in the Brentella Canal, with a total equivalent (TEQ) concentration of 2858 and 64,130 ng kg-1 at the surface and at 18-20 cm depth, respectively. High values can be also found in the Nord Industrial Canal and in the Salso Canal: 3000 and 2500 ng kg-1, respectively. Lagoon samples are much less contaminated, with TEQ downcore maxima of 22 and 53 ng kg-1, being influenced by pollutants stored in sediments of the canals only occasionally. Sediment chronology, based on 210Pb and 137Cs activity-depth profiles, shows that the highest fluxes occurred in the 1970s and early 1980s, and that pollution is now decreasing. Homologue profiles can distinguish PCDD/Fs from different sources: combustion for octachlorinated dibenzo-p-dioxin (OCDD) and stripping of VCM for octachlorinated dibenzofuran (OCDF) alone, whereas several activities carried out in the past within the industrial area can be responsible of the overall contamination by furans.     

Use of the clam Asaphis deflorata as a potential indicator of organochlorine bioaccumulation in Hong Kong coastal sediments

In order to test its ability as a bioaccumulator, the deposit feeding clam Asaphis deflorata, collected from a clean coastal area in Hong Kong (Ding Chau, Ma On Shan), was exposed to field-collected sediments representing a gradient of contamination. Sediments were collected from four sites, namely Shek O in the southeast of Hong Kong Island, Ap Lei Chau in Aberdeen district, Tai Kok Tsui in the Mong Kok waterfront, and To Kwa Wan near Kowloon City pier. Shek O was the cleanest and To Kwa Wan was the most polluted site. Replicate 30 L fiberglass tanks containing equal numbers of organisms were maintained for 28 days, with equal quantities of sediment and filtered seawater. Prior to this the animals were depurated for 10 days in clean sediment. During the 28-day holding period, sampling of both sediment and clams was performed on days 0, 10 and 28, along with condition index measurements. Concentrations of organochlorine pesticides and PCBs in sediments showed that Shek O was the cleanest of the sites, followed by Ap Lei Chau, Tai Kok Tsui and To Kwa Wan. There were marked differences among the sites, especially in PAH concentrations, with To Kwa Wan showing extremely high sediment concentrations.Following exposure, the biota concentrations of various organochlorine pesticides and PCBs suggested that there was sediment concentration-dependent accumulation by A. deflorata on day 10 of the experiment. However, there was no significant increase on day 28 for any of the compounds tested. Contaminant concentrations in clams feeding on Shek O sediments were minimal and almost the same on all sampling days, indicating the presence of only baseline contaminant concentrations. The condition indices (CI) of the clams on various sampling days showed that the animals were relatively more stable in sediment from Shek O. There were some deaths in all the test tanks (including those containing Shek O sediments). In addition, clams appeared to bury faster in the cleaner Shek O sediment and remained buried, unlike those in the sediment from the other locations, where some remained predominantly on the surface. Nevertheless, despite not completely burying, the clams in other test sediments still exhibited sediment concentration-dependent tissue accumulation. The results of the investigation indicate the possibility of developing a standard test protocol for bioaccumulation by sediment dwelling invertebrates, for practical use in Hong Kong, using A. deflorata.     

Cytochrome P450, acetylcholinesterase and gonadal histology for evaluating contaminant exposure levels in fishes from a highly eutrophic brackish ecosystem: the Orbetello Lagoon,Italy

Biochemical markers and ovarian histology were investigated in prespawning females of grass goby (Zosterisessor ophiocephalus) and grey mullet (Mugil cephalus) collected, respectively, in late spring and summer 2000 in four sites of a highly eutrophic brackish ecosystem of central Italy, the Orbetello Lagoon. Exposure to chlorinated and aromatic hydrocarbons was evaluated in fish livers by the somatic liver index (SLI) and by measuring 7-ethoxyresorufin-O-deethylase (EROD) and benzo(a)pyrene monooxygenase (BaPMO) activities. Acetylcholinesterase (AChE) activity was measured in brain and gills to evaluate exposure to organophosphates (OPs) and carbamates (CBs). The gonad somatic index (GSI) was used to confirm ovarian maturation and ovarian histology was investigated as a potential biomarker for environmental effects. Samples from the Western Basin, near a sewage treatment plant (STP) off the town of Orbetello, showed higher SLI values and higher EROD and BaPMO activities than those collected from the Ansedonia Canal (AC) in the Eastern Basin (p<0.05) and respect to those from reference sites: the Albegna River (AR) Delta for grass goby and the Nassa Canal (NC), connected with the sea, for grey mullet both located in the Western Basin as well. Low brain AChE activity was observed in both species from the reference sites (AR and NC) in association with the presence of anomalies in developing oocytes: unexpectedly small in grass goby and irregular disintegrated cytoplasm in grey mullet. The results indicate that the Western Basin is more polluted than the Eastern Basin particularly in the Orbetello where the sewage treatment plant may be a source of aromatic and chlorinated compounds while the Albegna River and the Nassa Canal may be sources of OPs and CBs.     

Bioaccumulation and biomarker responses of trace metals and micro-organic pollutants in mussels and fish from the Lagoon of Venice, Italy

The aims of this study were to evaluate levels and effects of trace metals (Cd, Cr, Cu, Fe, Mn, Pb, Zn), PAHs and PCBs in mussels and fish native to the Lagoon of Venice. To this end, bioaccumulation and various biomarkers of generic stress and exposure (malondialdehyde, MDA and metallothioneins, MT; ethoxyresorufin O-deethylase activity, EROD; fluorescent bile metabolites and somatic indices in fish; lysosomal membrane stability, LMS; lipofuscins, LF; neutral lipids, NL and survival-in-air, SOS in mussels) were measured in organisms seasonally collected from two differently influenced areas. In mussels, metal bioaccumulation levels at the two sites were low and fluctuated similarly, exhibiting decreasing levels in summer, like MDA and MT. Micro-organic pollutants and the probably related biological response of LMS did not show either significant site differences or seasonal trends. LF and NL revealed inconclusive patterns. The results of the survival-in-air test were quite erratic, showing that they were associated with the high variability of both natural and physiological parameters temperature, food, reproduction, and body reserve cycle. In fish, both inorganic and organic micropollutant levels were low, and differences were detected between sites only for PCBs. The pattern exhibited by micro-organic contaminants was clearly related to the reproductive cycle at both sites; a relationship with PCBs, EROD and MDA was established at the site where their levels were generally higher. Bile metabolites (3-OH benzo(a)pyrene, 1-OH pyrene) were associated with PAH contents.     

Detecting genotoxicity using the Comet assay following chronic exposure of Manila clam Tapes semidecussatus to polluted estuarine sediments

Sediments frequently cause damage to biota due to the accumulation of toxic compounds and the bioavailability of sediment-bound contaminants. Damage can be assessed using biomarkers, such as the degree of genotoxic impact following in vivo exposure to pollutants. Genotoxic damage, expressed as single-strand DNA breaks, was measured in cells isolated from haemolymph, gill and digestive gland from the clam Tapes semidecussatus, using the single cell gel electrophoresis (Comet assay). Clams were exposed for three weeks to sediment samples collected from a polluted site and a ‘clean’ reference site.

The level of DNA damage was assessed using an image analysis package and expressed as Tail Moment. Throughout the study, significant differences in DNA damage were recorded for each tissue type between clams exposed to the two sediment samples. We conclude that the Comet assay is a useful tool for the detection of DNA damage in clams chronically exposed to polluted sediments.     

POPs in edible clams from different Italian and European markets and possible human health risk

Human intake of persistent organic pollutants (POPs) occurs primarily through diet and fish and other seafood represent the principal means of contamination. Despite this, few legal limits have been established to protect human health from the various chemicals that are alleged to be carcinogenic or endocrine disruptor compounds.Several pools of edible clams (Tapes philippinarum and Venus gallina) bought in different Italian and European markets were examined in order to perform a risk assessment of some of these contaminants and also to identify possible health issues. POP concentrations in the soft tissues of mollusks varied considerably among source farming sites, particularly in the Venice Lagoon. Some POPs (DDTs, HCB and HCHs) showed very low concentrations and no risk for human health, in line with the limits established by European law. In contrast, PAHs could represent a potential hazard for human health and PCB levels in the most contaminated sites reached or even exceed the Minimal Risk Level as set by the ATSDR for human consumption, even if all the samples did not exceed the PCB limits in edible seafood as set by the FDA.     

Use of the clam Asaphis deflorata as a potential indicator of organochlorine bioaccumulation in Hong Kong coastal sediments

In order to test its ability as a bioaccumulator, the deposit feeding clam Asaphis deflorata, collected from a clean coastal area in Hong Kong (Ding Chau, Ma On Shan), was exposed to field-collected sediments representing a gradient of contamination. Sediments were collected from four sites, namely Shek O in the southeast of Hong Kong Island, Ap Lei Chau in Aberdeen district, Tai Kok Tsui in the Mong Kok waterfront, and To Kwa Wan near Kowloon City pier. Shek O was the cleanest and To Kwa Wan was the most polluted site. Replicate 30 L fiberglass tanks containing equal numbers of organisms were maintained for 28 days, with equal quantities of sediment and filtered seawater. Prior to this the animals were depurated for 10 days in clean sediment. During the 28-day holding period, sampling of both sediment and clams was performed on days 0, 10 and 28, along with condition index measurements. Concentrations of organochlorine pesticides and PCBs in sediments showed that Shek O was the cleanest of the sites, followed by Ap Lei Chau, Tai Kok Tsui and To Kwa Wan. There were marked differences among the sites, especially in PAH concentrations, with To Kwa Wan showing extremely high sediment concentrations.Following exposure, the biota concentrations of various organochlorine pesticides and PCBs suggested that there was sediment concentration-dependent accumulation by A. deflorata on day 10 of the experiment. However, there was no significant increase on day 28 for any of the compounds tested. Contaminant concentrations in clams feeding on Shek O sediments were minimal and almost the same on all sampling days, indicating the presence of only baseline contaminant concentrations. The condition indices (CI) of the clams on various sampling days showed that the animals were relatively more stable in sediment from Shek O. There were some deaths in all the test tanks (including those containing Shek O sediments). In addition, clams appeared to bury faster in the cleaner Shek O sediment and remained buried, unlike those in the sediment from the other locations, where some remained predominantly on the surface. Nevertheless, despite not completely burying, the clams in other test sediments still exhibited sediment concentration-dependent tissue accumulation. The results of the investigation indicate the possibility of developing a standard test protocol for bioaccumulation by sediment dwelling invertebrates, for practical use in Hong Kong, using A. deflorata.     

Total oxyradical scavenging capacity responses in Mytilus galloprovincialis transplanted into the Venice lagoon (Italy) to measure the biological impact of anthropogenic activities

Oxidative stress related investigations to monitor the impact of the pollutant discharges into the Venice lagoon (Italy) originating from anthropogenic activities (raw sewage water, agricultural and industrial effluents, oil tanker traffic), on marine organisms have classically been carried out by analyzing specific, single antioxidants (i.e. catalase, superoxide dismutase). In this paper, two studies are reported where the total oxyradical scavenging capacity assay (TOSC) was selected and measured toward peroxyl, and hydroxyl free radicals, and peroxynitrite in mussels (Mytilus galloprovincialis) transplanted into the Venice city and throughout the lagoon to measure the biological effects of anthropogenic activities. In the first experiment, mussels from a clean site (farm) were transplanted to the urban area of Venice for 0, 1, 2 and 4 weeks; cytosolic TOSC toward peroxyl and hydroxyl free radicals, and peroxynitrite revealed that the transplantation process caused a stress (handling stress, anoxia, oxidative burst) resulting in a reduction of TOSC in both control and urban sites, therefore, preventing clear interpretation of the data after one week. At week 2, a significant TOSC reduction (P < 0.05) toward peroxyl and hydroxyl radicals in the urban site revealed that mussels experienced oxidative pressure exerted by pollutants. Most TOSC values returned to initial levels at week 4; however, TOSC induction was noticed in the control group toward peroxyl and hydroxyl radicals while in the exposed group it was not indicating an inhibition of the oxidative metabolism. In the second experiment, mussels were deployed at seven different sites throughout the lagoon. After five weeks of exposure, significant TOSC reduction was measured (P < 0.05) toward peroxyl for Palude della Rosa, Chioggia and Valle Millecampi, toward hydroxyl radicals for Valle Millecampi and Campalto and toward peroxynitrite for Valle Millecampi. Although these data indicate a depletion of the low molecular weight scavengers, additional biomarkers are needed to draw a conclusion on the health of the mussels. TOSC was proved to be an interesting health index parameter to measure pollution impact in a transplantation study provided that the mussels are exposed for two weeks and a control is run in parallel.     

Esterases activities and lipid peroxidation levels in muscle tissue of the shanny Lipophrys pholis along several sites from the Portuguese Coast

This study is part of a project aiming to validate the use of the intertidal shanny Lipophorys pholis as a sentinel species in pollution monitoring in NW European marine ecosystems. To this end, a characterisation of acethylcholin (AChE), butyrylcholin (BChE) and propionylcholin (PrChE) esterases in L. pholis muscle was performed and the results indicated that AChE was predominant. Furthermore, the use of eserine sulphate and BW284c51 (0.64–800 μM), and iso-OMPA (0.08–16 mM), confirmed the measurement of true cholinesterases (ChEs) as well as the presence of pseudocholinesterases. The field application of these markers to L. pholis, sampled in seven locations along the Portuguese coast, revealed that fish were likely to be affected by neurotoxic compounds. This was indicated by the significant depletion of AChE (p < 0.05) in animals collected at urban and industrialised sites, compared with those from reference locations. The inclusion of a marker of effect, measured as lipid peroxidation levels in muscle tissue, also revealed the existence of site differences. Overall, the study further validates the utility of L. pholis in pollution monitoring studies.