Soft Tissues Of Mussels Research Articles

New evidences for old biomarkers: Effects of several xenobiotics on EROD and AChE activities in Zebra mussel ( Dreissena polymorpha)

The biomarker approach is widely used both in vertebrates and invertebrates for environmental biomonitoring, because it can supply an integrated response for multi-xenobiotics contamination. However, the use of biomarkers requires the identification of every possible variation that can influence the biochemical response, because ecosystems are generally subject to a mixture of pollutants, which can create additive, opposite or competitive effects. In recent years, there has been considerable interest in the use of biomarkers within marine bivalves, while very few data are available for freshwater molluscs. The aim of this research was to investigate changes on EROD and AChE activities in the freshwater bivalve Zebra mussel ( Dreissena polymorpha) exposed to different pollutants (Arochlor 1260, CB 153 and 126, pp’DDT, chlorpyrifos, carbaryl) at laboratory conditions, in order to standardize the analytical procedures and to highlight eventual interferences on enzyme activities. Chemical concentrations in the mussel soft tissues were analyzed by GC/MS-MS. Main results showed a significant induction of EROD activity when mussels were exposed to 100 ng/l of PCB mixture of Arochlor 1260 and dioxin-like CB 126, but this congener showed also a clear competitive inhibition after 48 h of exposure. Surprisingly, pp’DDT determined a significant decrease of basal EROD activity after only 24 h of exposure, even if it was not possible to discriminate between the effect of the parent compound and that of its metabolites (DDD, DDE). We also found an interaction between the organophosphate insecticide chlorpyrifos, which does not directly decrease the AChE activity, and terbutilazine. This herbicide increased the biotransformation of the organophosphate compound to its oxidized metabolite (oxon), a much stronger AChE inhibitor. The possible use of the oxime Pyridine-2-Aldoxime Methochloride (2-PAM) to bring back the catalytic activity to basal levels was also demonstrated.

Temporal and spatial trends in heavy metal concentrations in the marine mussel Mytilus edulis from the Western Scheldt estuary (The Netherlands)

Since the first North Sea Conference (1984, Bremen), all countries bordering the North Sea made commitments to reduce discharge of hazardous substances into the North Sea. From Belgium and The Netherlands, large reductions (upto 90) in heavy metal emissions from land-based sources have been reported between 1985 and 2000. Recently, some studies in the Western Scheldt estuary (WS) have shown that total metal concentration in the water, sediments and suspended particles have decreased compared to levels in the 70s. However, data on aquatic organisms is still very limited and it is therefore difficult to confirm whether the reductions in pollution input and generally improving water quality in the WS have a corresponding impact on the levels of heavy metals in aquatic organisms. The current study measured metal concentrations in the soft tissues of mussels, Mytilus edulis (known to be good indicators of environmental metal pollution) during the period 1996–2002. Spatial (salinity and pollution gradients), temporal and seasonal variations were also studied. Results showed a down-stream decreasing trend for the metals studied (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn) during all sampling campaigns. There was also a significant seasonal effect on tissue metal concentrations, with a peak observed around spring in both WS and the nearby less polluted Eastern Scheldt (ES). On temporal trends, a clear drop of metals in mussels was observed in the early 80s coinciding with the start of the efforts to reduce chemical pollution input into the North Sea. Since those early reductions, metal concentrations in mussels generally remained unchanged upto mid 90s. However, in recent times metal concentration in mussels have increased significantly, for example Cd in 2002 was almost 10 times the values in 1983 and similar to levels seen during the peak in the 70s. Other metals also increased in the 90s also reaching levels seen in the 70s. As there is no indication of recent increase in metal input into the estuary, we suggest that increased metal concentrations observed in mussels in recent years especially in the upper estuary are most likely a result of changes in physical and chemical speciation and metal bioavailability. Such changes may be caused by changes in some water quality parameters in the estuary (i.e. increased dissolved oxygen, concentration of organic matter), resulting in conditions that favour releases of sediment-bound metals into the water column. The relationship between metal content and season showed very similar annual profiles in the polluted WS and less polluted ES. Thus, seasonal variations in metal concentrations appear to be largely controlled by biological processes, while total body burdens are dependent on environmental levels and bioavailability.

Ultrasound-assisted solvent extraction of total polycyclic aromatic hydrocarbons from mussels followed by spectrofluorimetric determination

An ultrasound-assisted solvent extraction procedure has been optimised to speed up total polycyclic aromatic hydrocarbons (T-PAHs) extraction from mussel soft tissue. The T-PAHs releases have been evaluated by spectrofluorimetry (excitation and fluorescence emission wavelengths of 300 and 382 nm, respectively, and using chrysene as calibrant). Variables such as sonication time, ultrasound frequency, n-hexane volume, dichloromethane volume, number of repeated extractions with n-hexane and number of repeated extraction with dichloromethane were simultaneously studied by applying a Plackett–Burman design (PBD) approach. Results showed that ultrasound frequency and n-hexane and dichloromethane volumes were statistically significant variables (confidence interval of 95%). These last two variables were finally optimised by using central composite designs (CCD), yielding optimum n-hexane and dichloromethane volumes of 2.5 and 6.5 ml, respectively. The lowest T-PAHs releasing at high ultrasound frequency (35 kHz) led to choice the lowest ultrasound frequency (17 kHz) to perform the extraction. Variables such as sonication time and number of repeated extraction with n-hexane or dichloromethane were statistically non-significant and they were fixed at 10 min and the extraction with n-hexane and dichloromethane were performed once. The limit of detection was 0.021 μg g −1 (referred to dried mass), the repeatability of the overall method was 4.7% ( n = 9) and the analytical recoveries were between 98 and 105%. The proposed method was finally applied to 16 mussel samples ( Mytilus galloprovincialis) from Ría de Arousa estuary (Galicia, northwest Spain).

Modification of trace metal accumulation in the green mussel Perna viridis by exposure to Ag, Cu, and Zn

To examine the Cd, Hg, Ag, and Zn accumulation in the green mussel Perna viridis affected by previous exposure to Cu, Ag, or Zn, the dietary metal assimilation efficiency (AE) and the uptake rate from the dissolved phase were quantified. The mussel's filtration rate, metallothionein (MT) concentration, and metal tissue burden as well as the metal subcellular partitioning were also determined to illustrate the potential mechanisms underlying the influences caused by one metal pre-exposure on the bioaccumulation of the other metals. The green mussels were pre-exposed to Cu, Ag, or Zn for different periods (1–5 weeks) and the bioaccumulation of Cd, Hg, Ag, and Zn were concurrently determined. Pre-exposure to the three metals did not result in any significant increase in MT concentration in the green mussels. Ag concentration in the insoluble fraction increased with increasing Ag exposure period and Ag ambient concentration. Our data indicated that Cd assimilation were not influenced by the mussel's pre-exposure to the three metals (Cu, Ag, and Zn), but its dissolved uptake was depressed by Ag and Zn exposure. Although Hg assimilation from food was not affected by the metal pre-exposure, its influx rate from solution was generally inhibited by the exposure to Cu, Ag, and Zn. Ag bioaccumulation was affected the most obviously, in which its AE increased with increasing Ag tissue concentration, and its dissolved uptake decreased with increasing tissue concentrations of Ag and Cu. As an essential metal, Zn bioaccumulation remained relatively stable following the metal pre-exposure, suggesting the regulatory ability of Zn uptake in the mussels. Zn AE was not affected by metal pre-exposure, but its dissolved uptake was depressed by Ag and Zn pre-exposure. All these results indicated that the influences of one metal pre-exposure on the bioaccumulation of other metals were metal-specific due to the differential binding and toxicity of metals to the mussels. Such factors should be considered in using metal concentrations in mussel's soft tissues to evaluate the metal pollution in coastal waters.

Use of enzymatic hydrolysis for the multi-element determination in mussel soft tissue by inductively coupled plasma-atomic emission spectrometry

A systematic evaluation of different variables affecting the enzymatic hydrolysis of mussel soft tissue by five enzymes, three proteases (pepsin, pancreatin and trypsin), lipase and amylase, has been carried out for the determination of trace elements (As, Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) by inductively coupled plasma-atomic emission spectrometry (ICP-AES). Enzymatic hydrolysis methods offers advantages such as a less species alteration, safer laboratory conditions and a less contaminant wastes. The enzymatic hydrolysis was performed in an incubation camera Boxcult with orbital and horizontal shaker. Variables affecting the enzymatic hydrolysis process were simultaneously studied by applying a Plackett–Burman design (PBD). For a confidence interval of 95%, the significant factors for all enzymes and for most of the elements were the pH, the incubation temperature and the ionic strength. These significant factors were optimized later by using a central composite design (CCD), which gave optimum conditions at pH of 1, incubation temperature of 37°C and ionic strength fixed by sodium chloride at 0.2M when using pepsin. For pancreatin, trypsin, lipase and amylase there were found two different optimum condition sets. The first one involves the use of a 0.5M phosphate buffer (ionic strength), at a pH of 6 and at an incubation temperature of 37°C, which allows the quantitative extraction of Al, Cr, Mn, Pb and Zn. The second conditions set employees a 0.1M phosphate buffer (ionic strength), a pH of 9 and an incubation temperature at 37°C, and it results adequate to extract As, Cd, Cu, Fe and Ni. Analytical performances, repeatability of the over-all procedure and accuracy, by analyzing DORM-1, DORM-2 and TORT-1 certified reference materials, were finally assessed for each enzyme. Good agreement with certified values has been assessed for most of the elements (As, Cd, Cr, Cu, Mn, Ni, Pb and Zn) when using trypsin, pepsin and/or pancreatin, except for Cd and Pb in DORM-1 and DORM-2 because of the certified contents in such certified reference materials are lower than the limit of detection (0.10 and 0.16μgg−1 for Cd and Pb, respectively, for the use of trypsin).

Use of enzymatic hydrolysis for the multi-element determination in mussel soft tissue by inductively coupled plasma-atomic emission spectrometry

A systematic evaluation of different variables affecting the enzymatic hydrolysis of mussel soft tissue by five enzymes, three proteases (pepsin, pancreatin and trypsin), lipase and amylase, has been carried out for the determination of trace elements (As, Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) by inductively coupled plasma-atomic emission spectrometry (ICP-AES). Enzymatic hydrolysis methods offers advantages such as a less species alteration, safer laboratory conditions and a less contaminant wastes. The enzymatic hydrolysis was performed in an incubation camera Boxcult with orbital and horizontal shaker. Variables affecting the enzymatic hydrolysis process were simultaneously studied by applying a Plackett–Burman design (PBD). For a confidence interval of 95%, the significant factors for all enzymes and for most of the elements were the pH, the incubation temperature and the ionic strength. These significant factors were optimized later by using a central composite design (CCD), which gave optimum conditions at pH of 1, incubation temperature of 37 °C and ionic strength fixed by sodium chloride at 0.2 M when using pepsin. For pancreatin, trypsin, lipase and amylase there were found two different optimum condition sets. The first one involves the use of a 0.5 M phosphate buffer (ionic strength), at a pH of 6 and at an incubation temperature of 37 °C, which allows the quantitative extraction of Al, Cr, Mn, Pb and Zn. The second conditions set employees a 0.1 M phosphate buffer (ionic strength), a pH of 9 and an incubation temperature at 37 °C, and it results adequate to extract As, Cd, Cu, Fe and Ni. Analytical performances, repeatability of the over-all procedure and accuracy, by analyzing DORM-1, DORM-2 and TORT-1 certified reference materials, were finally assessed for each enzyme. Good agreement with certified values has been assessed for most of the elements (As, Cd, Cr, Cu, Mn, Ni, Pb and Zn) when using trypsin, pepsin and/or pancreatin, except for Cd and Pb in DORM-1 and DORM-2 because of the certified contents in such certified reference materials are lower than the limit of detection (0.10 and 0.16 μg g−1 for Cd and Pb, respectively, for the use of trypsin).

In vivo exposure of Dreissena polymorpha mussels to the quinones menadione and lawsone: menadione is more toxic to mussels than lawsone.

The principal aim of this study was to assess whether the two quinones, menadione (2-methyl-1,4-naphthoquinone) and lawsone (2-hydroxy-1,4-naphthoquinone), elicit differential toxicity in mussels as has been reported for higher organisms. Therefore, the effects of short-term (48 h) and long-term (20 days) exposure of the two quinones at concentrations of 0.56 and 1 mg l(-1) to zebra mussels, Dreissena polymorpha, under laboratory conditions were studied. After the short-term exposure, the specific activities of the two-electron quinone oxidoreductase (DT-diaphorase) and the one-electron catalysing quinone reductases NADPH-cytochrome c reductase and NADH-cytochrome c reductase were determined in the gills and the rest of the soft tissues (soft mussel tissues minus the gills) of both treated and control mussels. At the higher concentrations of menadione and lawsone used, a significant reduction of the activity of NADPH-cytochrome c reductase in the gills and in the rest of the soft mussel tissues (by 33-34% and 31-43%, respectively) was observed. The activities of DT-diaphorase and NADH-cytochrome c reductase were not significantly affected. Interestingly, DT-diaphorase was observed in the gills, an organ requiring protection against antioxidants. Furthermore, a single-cell electrophoretic assay (comet assay) performed with gill cells to assess DNA damage by the quinones did not show any significant difference between the treated and the control organisms. This indicates that the formation of reactive species by the quinone metabolism in vivo in the mussels was possibly suppressed through the concerted action of DT-diaphorase and antioxidant enzymes. The results of in vitro experiments with gill extracts confirmed the protective role of DT-diaphorase. The rate of the two-electron quinone reduction was found to be five times that of the one-electron quinone reduction. The results of the long-term exposure unambiguously demonstrated that in mussels menadione, unlike in higher organisms, is more toxic than lawsone. The lack of detectability of xanthine oxidase in the mussel tissues could explain the comparatively lower toxicity of lawsone in the invertebtrate, lending support to a previous suggestion that xanthine oxidase might be responsible for the mechanism of toxicity of lawsone in higher organisms in vivo.

Distribution and coassociations of trace elements in soft tissue and byssus of Mytilus galloprovincialis relative to the surrounding seawater and suspended matter of the southern part of the Korean Peninsula

The concentrations of Cd, Co, Cu, Cr, Fe, Hg, Mn, Ni, Pb, Sn, Ti and Zn were analyzed by AAS, ICP MS and AFS in soft tissues and byssal threads of Mytilus galloprovincialis from Masan Bay and Ulsan Bay, Korea. Spatial variations in metal concentrations were found. The levels of Cd, Pb, Hg, Cu, Zn, Co and Mn were very high in the mussels from Ulsan Bay (Sts. U1, U2) and comparable with elevated concentrations of these elements in Mytilus sp. reported to date for other geographical areas. Seasonal differences in some metal concentrations were also observed. These variations may be caused by factors such as: a large difference in seawater temperature, food supply for the mussel population and/or freshwater runoff of particulate metal to the coastal water and weight changes brought about by gonadal development and the release of sexual products. Pb, Cu, Zn, Co, Ni, Fe and Mn were more enriched in byssal threads than in the soft tissues, hence the byssus seems to be more sensitive in reflecting the availabilities of trace metals in the ambient waters. Concentrations of trace metals varied with respect to the size of mussels and season, depending on many factors like sexual development, and seawater temperature, etc. The levels of some trace metals in seawater, especially in suspended matter were correlated significantly with those in soft tissues and byssal threads. There were spatial variations in metal concentrations in the soft tissue and byssus attributed to different sources of trace elements located near the sampling sites. There were significant relationships between concentrations of some metals (Cd, Cu, Pb, and Zn) in mussel soft tissues and byssal threads and suspended matter. This suggests that M. galloprovincialis can be used as a sensitive biomonitor for the availabilities of trace elements in the coastal waters off Korea.

Are coastal resources of NW Portugal fingerprinting hydrocarbons released from the Prestige accident?

Seventy-one samples of mussels, sardine, hake, blue whiting and pouting from the NW Portuguese coast, collected between 18 November 2002 and 6 January 2003, were analyzed for total hydrocarbons, individual n-alkanes and major isoprenoids, pristane and phytane. Hydrocarbons (expressed on chrysene equivalents) registered in whole soft tissues of mussels were below 0.5 µg g–1, with a few exceptions (maximum 1.6 µg g–1). Relatively low levels and sporadic enhanced values were also found in fish fillet samples. In spite of this, analyses of individual n-alkanes and major isoprenoids showed that 10–61% of the samples (according to the species) exhibited these compounds in the proportions of the Prestige fuel signature. Similar diagnostic ratios in biological samples and Prestige fuel and n-alkane composition corroborate the fingerprint hypothesis in coastal resources from NW Portugal.

Metal concentrations in mussel Crenomytilus grayanus and oyster Crassostrea gigas in relation to contamination of ambient sediments

The relationship between Zn, Cu, Cd, Pb, and Ni concentrations in soft tissues of mussels and oysters and those in ambient sediments was examined. The study area was the coastal habitats of the northwestern Sea of Japan, which has a broad range of contamination due to urban sewage. Significant increases of all metals, except Ni, were observed in mussel Crenomytilus grayanus at concentrations of easily leachable metals in ambient sediments higher than 2, 100, and 800 μg/g for Cd, Cu, and Zn, respectively. Pb was accumulated by the mussels with no distinct threshold in Pb contamination of sediments. After 2 months, C. grayanus transplanted from a pristine to a contaminated locality increased in Pb up to 12%, and Cu up to 68% of concentrations of these metals in the contaminated local mussels, but Zn and Cd concentrations increased only in the excretory tissue, not in the somatic tissue, or in the soft tissue as a whole. Such regulation of metal accumulation puts limitations on the use of C. grayanus for monitoring of slightly and moderately contaminated localities. On the other hand, the oyster Crassostrea gigas showed definite accumulation of all metals, except Ni, at moderate contamination of ambient sediments. As contamination of sediments increased, increases of Pb, Zn and Cu concentrations in C. gigas slowed, possibly due to physiological control at very high metal concentrations in oyster soft tissue. Thus, the mussel C. grayanus should be used mainly for the monitoring of heavy contaminated localities, and the oyster C. gigas is more suitable as an indicator of low and moderate contamination.

Biological availability of traffic‐related platinum‐group elements (palladium, platinum, and rhodium) and other metals to the zebra mussel (Dreissena polymorpha) in water containing road dust

The uptake and bioaccumulation of 15 road dust metals by the zebra mussel (Dreissena polymorpha) were investigated in laboratory exposure studies with emphasis on the traffic-related platinum-group elements (PGEs) palladium (Pd), platinum (Pt), and rhodium (Rh). The biological availability of the metals may depend on water characteristics, so the mussels were maintained in two types of water: nonchlorinated tap water and humic water of a bog lake, both of which contained dust of a moderately frequented road. After an exposure period of 26 weeks, soft tissues of the mussels were freeze-dried and analyzed for the metals. The metal concentrations in the mussel soft tissue ranged from several hundred micrograms per gram (e.g., for iron [Fe]) to less than 10 ng/g (for PGEs). Metal uptake from the road dust by the mussels was found for the PGEs and silver (Ag), bismuth (Bi), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), Fe, lead (Pb), and antimony (Sb). After maintenance of mussels in road dust-contaminated tap water, bioaccumulation factors (BAF = (C(exposed mussels) - C(control mussels))/C(total metal, water), where c is concentration) decreased in the following order: Cu > Cd > Ag > Pd > Sb > Pb > Fe > Pt > Rh. The biological availability of most metals was enhanced by humic water as compared to tap water. Our results show a hitherto unrecognized high availability of Pd for the mussels. Thus, this metal should be monitored more intensively in the environment to assess its distribution in the biosphere.

Implications from a field study regarding the relationship between polycyclic aromatic hydrocarbons and glutathione S-transferase activity in mussels

An aluminium smelter on the west coast of Scotland discharges an aqueous effluent containing polycyclic aromatic hydrocarbons (PAHs) at the head of Loch Leven. The loch also supports two mussel (Mytilus edulis) farms. Data are presented on burdens of PAHs in the soft tissues of mussels and the effect of these contaminants on glutathione S-transferase (GST) activity in mussel hepatopancreas. GST activity is shown to be correlated with total PAH burden and also with the concentrations of certain individual PAHs. These field data show that high molecular weight PAHs are closely correlated to GST activity, whereas low molecular weight PAHs are not. This suggests that 5- and 6-ring PAHs have a more pronounced role than 2- to 4-ring compounds in inducing GST activity in mussels from Loch Leven. It is proposed that it may be more appropriate to link GST activity with 5- and 6-ring compounds only, rather than with the total PAH burden.

Metallothionein concentration in the mussel Mytilus galloprovincialis as a biomarker of response to metal contamination: validation in the field

Mussels were translocated from a shell-fish breeding area (Sète, on the French Mediterranean coast) to sites exposed to trace element inputs in April 2000. They were recovered 3 months later. Whole soft tissues from all of the sites (n = 97) were analysed for arsenic, cadmium, chromium, copper, mercury, nickel, lead and zinc. Metallothioneins (MTs) were also measured in the digestive gland and in the remaining tissues (allowing calculation of whole soft tissue concentrations) at 22 of the 97 sites. MT concentrations in the digestive gland and the whole soft tissues were strongly correlated. The condition index varied with food availability at different sites. This did not influenced MT concentrations in the whole soft tissues, whereas the condition index was negatively correlated to trace element concentrations. A model is proposed to minimize this influence of condition. Metal concentrations adjusted using this model showed significant correlations with MT levels for those metals (cadmium, copper, nickel and zinc) that are known to bind to this protein, with the exception of mercury. Even in moderately contaminated sites, measurement of the MT level in the soft tissues of mussels was generally able to discriminate between different levels of contamination, allowing the use of a simplified procedure compared with dissection of the digestive gland. It is recommended to avoid translocation and sampling during the reproductive period, which is well documented for commercial species such as Mytilus sp.

Heavy metals and arsenic in sediments, mussels and marine water from Murano (Venice, Italy)

This paper describes a study of the trace element distribution in sediments, marine water and mussel Mytilus galloprovincialis of the Venetian Lagoon around the Island of Murano, an island with a long tradition of glassmaking. Trace elements analysed include Fe, Mn, Zn, Cu, Cr, Pb, Ni, Ag and As. Sediments are contaminated with Zn, Cu, Ag, As and Pb, with levels in the <2 m fraction that are likely to cause adverse biological effects to marine organisms. The pelite (<63 μm) is the main carrier of heavy metals at most sites. However, the fine-grained and coarse sand on the southern coast of Murano accounts for a significant proportion of Fe, Mn, Zn, Cr, Ag, Ni and Pb. Most trace element concentrations found in soft tissue of mussels appear to be within recommended Italian and international guidelines for shellfish for human consumption, the only exception being relatively high As levels. The bioaccumulation of Ag and Cr is more pronounced in the shell of these organisms. In the marine water of the lagoon, trace elements are more enriched than in other areas of the Mediterranean, with particular reference to the dissolved labile species of Zn, Mn, As, Cu, Ni and Cr.

Trace analysis of platinum in biological samples: a comparison between sector field ICP-MS and adsorptive cathodic stripping voltammetry following different digestion procedures

A study was carried out to analyse Pt trace levels in fish liver and mussel soft tissue, using two different analytical techniques. Sector field inductively coupled plasma mass spectrometry (SF-ICP-MS) combined with microwave digestion was compared with adsorptive cathodic stripping voltammetry (ACSV) following high pressure ashing (HPA). A microwave digestion procedure for animal tissue was developed, using HNO 3 and H 2O 2. The procedural detection limit of Pt with SF-ICP-MS in combination with this digestion procedure was 2.4 ng/l in the solution. The subsequent detection limit considering 300 mg liver tissue (wet weight) was found to be 200 pg/g Pt. In comparison, the detection power of ACSV after HPA digestion was about 20 times more sensitive. The higher procedural detection limit of the ICP-MS-based method is due to the high dilution necessary to reduce acid concentration. Nevertheless, the results of both analytical methods differed between 16 and 30% for Pt concentrations below 1 ng/g, and were almost identical (2–10% deviation) above this value. The Pt levels in the animal tissues analysed ranged between 0.1 and 2.3 ng/g.

Time-dependent accumulation of cyanobacterial hepatotoxins in flounders (Platichthys flesus) and mussels (Mytilus edulis) from the northern Baltic Sea.

There is only limited information about the accumulation of algal toxins in aquatic organisms in the Baltic Sea. In this study we measured total cyanobacterial hepatotoxin levels in blue mussel (Mytilus edulis) and flounderi (Platichthys flesus) tissues. Flounder were caught with gillnets from the western Gulf of Finland during July and August 1999. Blue mussels were collected from an enclosure at 3 m depth and from an artificial reef (wreck, 25-35 m depth) in the western Gulf of Finland between June and September 1999. Flounder liver and muscle samples and soft tissues of mussels were analyzed for the cyanobacterial hepatotoxins (nodularin, NODLN and/or microcystins, MCs) using an enzyme-linked immunosorbent assay (ELISA). Results showed a time-dependent accumulation of hepatotoxins in flounder and mussels. In flounder, the maximum concentration 399 +/- 5 (sd) ng NODLN or MC/g dry weight (dw) was found in the liver of specimens caught on 21 August 1999. No hepatotoxins were detected in muscle samples. The maximum concentration of 2150 ng +/- 60 (sd) ng hepatotoxin/g dw was found in the mussel soft tissues collected on 20 August 1999. Temporal NODLN or MC trends indicated depuration of cyanobacterial hepatotoxin from mussels at surface level and an increase in NODLN or MC concentrations in those from the sea bed. These studies showed that despite the low cyanobacteria cell numbers the cyanobacterial hepatotoxins can accumulate in flounder and mussels. This may allow the further transfer of cyanobacterial hepatotoxins in the food web.

The Effects of Sponge (Porifera) Biofouling on Zebra Mussel (Dreissena polymorpha) Fitness: Reduction of Glycogen, Tissue Loss, and Mortality

ABSTRACT The effects of freshwater sponge (Porifera) overgrowth on the fitness of adult zebra mussels (Dreissena polymorpha) were examined in East Chicago and Michigan City, Indiana harbors in southern Lake Michigan. There was significant reduction in glycogen content of sponge-covered zebra mussel soft tissue wet weight (mean=2.6%) when compared to the non sponge-covered population (mean=3.9%). Similarly, the soft tissue wet weight of sponge-covered zebra mussels (15 to 25 mm size class) was lower when compared to non-covered mussels. A comparison of the live:dead ratios of zebra mussels in both populations showed overgrowth by sponges had a significant lethal effect. These data suggest that the epibiont growth of freshwater sponges on adult zebra mussel shells negatively impacts the growth and survivability of zebra mussels.

Toxic and nutrient element concentrations in soft tissues of zebra and quagga mussels from Lakes Erie and Ontario

Zebra and quagga mussels were collected from Lakes Erie and Ontario in 1997 and the soft mussel tissues were analyzed for Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Se, Sr, V and Zn. No consistent relationships were apparent when comparing element concentrations in soft mussel tissues and mussel type, size range or sampling location. Literature dealing with the absorption of metals by both mussel types is reviewed.

Organochlorines in Greenland marine fish, mussels and sediments

Shorthorn sculpin ( Myoxocephalus scorpius), polar cod ( Boreogadus saida), blue mussels ( Mytilus edulis) and sediments were sampled in Greenland 1994–1995 at three locations at the west coast and one at the east coast. Fish liver, mussel soft tissue and sediments were analysed for PCBs (10 congeners), DDTs ( pp′), HCHs (α, β, γ), HCB and trans-nonachlor. The overall geometric mean concentrations found for PCBs were 17 μg kg −1 wet wt. in shorthorn sculpin liver, 33 μg kg −1 wet wt. in polar cod liver, and 0.86 μg kg −1 wet wt. in blue mussels. For the three species, the geometric mean concentrations for DDTs were 11, 36, and 0.39 μg kg −1 wet wt., respectively; for HCHs: 8.7, 32 and 0.56 μg kg −1 wet wt., respectively; for HCB: 4.2, 11 and 0.06 μg kg −1 wet wt., respectively; and for trans-nonachlor: 6.3, 19 and 0.16 μg kg −1 wet wt., respectively. All organochlorines in the sediment samples were below the detection limit of 0.1 μg kg −1 dry weight. For sculpins and mussels, most organochlorine compounds were found to increase with increasing lipid content. The weight of mussels did not influence organochlorine concentrations, whereas organochlorine content in general increased with fish length of sculpins. The concentrations were found to be comparable to levels in other Arctic regions, but orders of magnitude lower than levels found in the southern part of the North Sea. Organochlorine concentrations in sculpins showed a decreasing trend following the ocean current flowing from north to south at the east coast and from south towards north at the west coast of Greenland. The proportion of higher chlorinated PCBs (Cl atoms ≥6) in sculpin liver followed the decreasing trend of PCB concentrations.

Toxicological aspects associated with the ecology of Donax trunculus (Bivalvia, Mollusca) in a polluted environment

This study provides data on the ecology and toxicology observed in the population of Donax trunculus, a sand dwelling mussel, in the shallow subtidal of Haifa Bay (Mediterranean Sea, Israel). The studied population of the mussel forms a dominant fraction in a community of sand-dwelling molluscs in a zone located 5–25 m from the shoreline, and at depths of 20–120 cm, numbering up to 2000 per m2. Samples of the mussel were collected from three sites, located within 9 km of shore in Haifa Bay. These included a clean site (Akko), a site polluted by a chemical PVC-polymer industry (Frutarom), and a site polluted with oil and waste from the petrochemical industry (Qiryat Yam). Metal analysis indicated site-dependent variations in levels of cadmium, lead, copper and mercury in the mussel soft tissues. Copper levels were similar in the bivalves collected from all the sites. Levels of mercury were significantly higher in specimens from the PVC-polluted site (Frutarom) while levels of cadmium were higher in Akko and Qiryat Yam compared to Frutarom. Lead residues were found only in Donax from Akko. The residual contents of mercury, copper and cadmium were relatively high in the young and noticeably low in Donax of medium body size. The main site of deposition of metals was in the soft tissues of the bivalve, but bioaccumulation of metals was also found in the shells. A marked increase in permeability of gills and mantle to the fluorescent anionic dye — fluorescein (FLU) was detected in Donax from Qiryat Yam and especially Frutarom, compared to the bivalves sampled from Akko. Multiple foci of enhanced permeability (multiple fluorescent spots) were detected in all the individuals sampled from Frutarom but none in the bivalve samples from Akko. Lysosomal accumulation of the metachromatic fluorescent cationic probe, acridine orange — (AO), was significantly decreased in the tissues of D. trunculus from polluted sites, especially polluted by the PVC factory. This decrease correlated with lysosomal enlargement and the formation of secondary lysosomes. D. trunculus appears to possess the most effective biochemical and physiological defense mechanisms enabling it to survive in habitats of polluted shallow waters, where other sand dwelling mollusc species were absent or found only in waters deeper then 2.5 m.