Reduced survival and condition of the bivalves Protothaca staminea and Mytilus edulis buried by decomposing bark

The bivalves Protothaca staminea and Mytilus edulis were exposed to five depths of decomposing bark from an active log transfer facility for 13–96 days in a field experiment at Auke Bay, Alaska. Survival and condition of both species was inversely related to depth and duration of bark coverage. As little as 6 cm of bark reduced survival; survival declined most rapidly between 10 and 15 cm of bark. The depths of bark under which half the animals died after 96 days of exposure were 12·8 ± 1·6 cm for P. staminea and 10·9 ± 1·5 cm for M. edulis. Interstitial water in the bark deposits had low concentrations of dissolved O₂ (2·5–5·3 mg/liter) and elevated concentrations of hydrogen sulfide (1·0–8·1 mg/liter total sulfide) and ammonia (46–131 μg-at N/liter) compared to the surface water from the control treatment. Survival of P. staminea and M. edulis was inversely related to concentrations of hydrogen sulfide and ammonia and directly related to dissolved O₂ concentrations.     

Characterisation of choline esterases and their tissue and subcellular distribution in mussel (Mytilus edulis)

Acetylcholinesterase in mussel is potentially a useful biomarker of exposure to organophosphates (OP) in the marine environment. This study looked at cholinesterase activity in subcellular fractions of various tissues from the common mussel, Mytilus edulis. Measurement of enzyme rates demonstrated that although highest specific activity was found in foot 'mitochondrial' fraction, recovery of activity was very low. Gill 'microsomal' fraction had the second highest specific activity with a useful level of recovery and therefore was the most suitable tissue fraction for biomarker applications. Comparative studies of alternative alkylthiocholine substrates and competitive inhibitors suggest there is a single cholinesterase enzyme type present in this fraction. Inhibition of alkylcholine hydrolysis by BW284C51, specific to acetylcholinesterase in vertebrates, showed that cholinesterase activity in gill 'microsomal' fraction is inhibited by this compound but to a lesser extent than in vertebrate AChE. Inhibition of cholinesterase activity by azamethiphos in gill 'microsomal' fraction gave an IC50 of approximately 100 microM and showed both time and concentration dependence. However this indicates a lower potency compared to other animals and it is debatable whether mussel cholinesterase activity is useful as a biomarker of exposure in the field.     

Recent trends in organochlorine residues in mussels (Mytilus edulis) from the Mersey Estuary.

Blue mussels (Mytilus edulis) taken from several sites in the Mersey Estuary, an urban-industrial water body in NW England, have been analyzed for residues of the persistent organochlorines, DDT, PCB congeners nos. 28, 52, 101, 118, 138, 153 and 180 (ICES7), and alpha-MHCH (alpha-methyl hexachlorocyclohexane). The concentration range for sigmaPCB (ICES7) was 13.9-34.9 microg kg(-1) in 1994 compared with 9.6-31.9 microg kg(-1) in 1998, whilst the equivalent concentrations of sigmaDDT were 8.9-32.4 microg kg(-1) and 4.5-16.8 microg kg(-1), respectively. The concentration of alpha-MHCH, an organochlorine largely restricted to the Mersey Estuary, was 1.2-11 microg kg(-1) in 1994 and 0.3-1.3 microg kg(-1) in 1998. At both sampling dates the inner estuary sites of Rock Ferry and Egremont were more contaminated than the outer estuary sites of New Brighton. Dove Point and Caldy Blacks. The lower concentrations of organochlorines in mussels in 1998 compared with 1994 are thought to be early evidence of significant improvements in water quality derived from major capital investment in the treatment of industrial effluents and sewage wastewater, linked to a combination of stricter legislative controls and the environmental benefits from the adoption of new, clean technologies in manufacturing.     

Measurement of stress effects (scope for growth) and contaminant levels in mussels (Mytilus edulis) collected from the Irish Sea

The objective of this research was to quantify the impact of pollution along the coastlines of the Irish Sea. Pollution assessment was based on the combined measurement of scope for growth (SFG), and chemical contaminants in the tissues of mussels (Mytilus edulis) collected from 38 coastal sites around the Irish Sea during June-July in 1996 and 1997. On the UK mainland coast, the SFG showed a general trend with a significant decline in water quality in the Liverpool and Morecambe Bay region. High water quality was recorded along the west coast of Wales, as well as southwest England and northwest Scotland (clean reference sites outside the Irish Sea). Along the coast of Ireland there was a similar trend with reduced SFG within the Irish Sea region. SFG was generally low north of Duncannon and then improved north of Belfast. The poor water quality on both sides of the Irish Sea is consistent with the prevailing hydrodynamics and the spatial distribution of contaminants associated with urban/ industrial development. The decline in SFG of mussels on both sides of the Irish Sea was associated with a general increase in contaminant levels in the mussels. Certain contaminants, including PAHs, TBT, sigmaDDT, Dieldrin, gamma-HCH, PCBs, and a few of the metals (Cd, Se, Ag, Pb), showed elevated concentrations. Many of these contaminants were particularly elevated in the coastal margins of Liverpool Bay, Morecambe Bay and Dublin Bay. A quantitative toxicological interpretation (QTI) of the combined tissue residue chemistry and SFG measurements indicated that at the majority of coastal sites, c. 50 to > 80% of the observed decline in SFG was due to PAHs as a result of fossil fuel combustion and oil spills. TBT levels were highest at major ports and harbours, but these concentrations only made a minor contribution to the overall reduction in SFG. At no sites were individual metals accumulated to concentrations that could cause a significant effect on SFG. The study identified many sites where the observed reduction in SFG was far greater than predicted from the limited number of chemical contaminants analysed, thus indicating the presence of additional 'unknown toxicants'. Sewage (containing domestic, agricultural and industrial components) appears to be an important contributor to reduced SFG and linear alkylbenzenes (LABs) and As may provide suitable 'sewage markers'. There was a highly significant positive correlation between SFG and As (P < 0.001). This relationship may be due to reduced As uptake by algal food material and mussels at sites with elevated P04 concentrations (e.g. at sites with sewage inputs). Phosphate is a known competitive inhibitor of As accumulation, at least in algae. The results highlight that further research is required on 'sewage markers' in mussels. The SFG approach therefore provides a rapid, cost-effective and quantitative measure of pollution impact, as well as a means of identifying the causes through a QTI of tissue contaminants levels. It also serves to identify the presence of unidentified toxicants and areas that require further study.     

The effect of pollution on scope for growth of the mussel (Mytilus edulis L.) in Iceland

In order to evaluate effects of contaminants at high latitudes, the effects on scope for growth (SFG) were evaluated for both transplanted and resident blue mussel (Mytilus edulis L.) near large and small harbours in Iceland. Low SFG values were found for mussels at the mouth and in the inner part of the large Reykjavík harbour, which could be attributed to elevated levels of organotins and polyaromatic hydrocarbons. Mussels in smaller and/or less polluted harbours and sites off Reykjavík harbour only showed a slight reduction in SFG. No major effects were found at sites >3 km from the large harbour of Reykjavík. SFG provides valuable information on the effects of harbour pollutants in Iceland, in particular in areas of extensive shipping activity. The use of this biomarker on high latitude organisms is highly relevant due to extended oil exploration at higher latitudes.     

厚壳贻贝性腺不同发育时期肥满度与生化成分分析

为研究厚壳贻贝性腺发育不同时期的营养物质积累和消耗规律,本文分析测定了3个年龄组的厚壳贻贝在性腺发育5个时期的肥满度、含水量及生化成分。结果表明:(1)相同发育时期的肥满度和含水量组间差异不显著(P＞0.05),但在不同发育时期存在显著差异(P＜0.05);成熟期的肥满度最高,为17.76%~19.21%,约为休止期最低时的3.15倍;含水量则正好相反,以休止期最高,为93.29%~95.32%,约为成熟期最低时的1.15倍。(2)总糖、粗脂肪、粗蛋白和灰分在不同发育时期均有显著差异(P＜0.05),粗蛋白和粗脂肪含量在成熟期达到峰值,粗蛋白为72.01%,粗脂肪为13.52%,两者分别在休止期和增殖期最低;总糖含量在排放期最高为13.56%,灰分含量以休止期最高为7.59%,两者均以成熟期含量最低。(3)蛋白质的积累与消减正好与肥满度的变化相吻合,证实厚壳贻贝在性腺发育过程中吸收并贮存大量营养物质,使肥满度逐渐升高,随配子的排放肥满度急剧下降。     

Multidrug resistance in the embryos and larvae of the mussel Mytilus edulis.

Cells exhibiting the multidrug resistance (MDR) phenotype demonstrate a decreased intracellular drug accumulation due to an active outward transport and decreased intracellular flux. This study demonstrates the inhibition of MDR in mussel (Mytilus edulis) embryos and larvae based on a simple bioassay. The development of embryos was assessed and abnormalities identified at key stages of development, including gastrulation, trochophore and prodissoconch stages. The incidence of developmental abnormalities was significantly increased in the presence of vinblastine, MMS, chloroquine, mitomycin-C, cadmium chloride and colchicine, compared to clean seawater. Consistently, there was a further increase in the number and severity of deformities observed when each toxin was added in the presence of verapamil. Larval growth was also significantly impaired in the presence of verapamil. Increased accumulation of fluorescent MDR dyes, such as rhodamine B, has been measured and shown to be verapamil sensitive. This bioassay encompasses a period of intense cellular activity during which the impairment of a number of critical processes results in abnormal growth and development.     

Cadmium speciation and transport in the blood of the bivalve Mytilus edulis.

Cadmium (Cd) speciation in the blood plasma of Mytilus edulis was investigated using the metal speciation model MINTEQA2. In the presence of inorganic ions alone, Cd-chloro complexes dominated the speciation (97% of total Cd), with 3% as Cd2+. Inclusion of a novel Cd-binding histidine-rich glycoprotein (HRG) purified from mussel blood plasma decreased the contribution of chloro-complexes to 11.9%, with 86.8% of the Cd bound to the HRG and 1.3% present as Cd2+. Cd transfer from the blood plasma to the kidneys in vivo was studied by injecting 109Cd (both with and without additional chelation) into mussels. Oxine and EDTA complexed a significant amount of blood-borne Cd (23.7% Cd Oxine; 57.1% CdEDTA). In the presence of each chelator, plasma retained significantly more Cd, although there was no significant difference in Cd uptake by tissues (kidney, gill-mantle, and remaining viscera).     

Immuno-localisations (GSSP) of subcellular accumulation sites of phenanthrene, aroclor 1254 and lead (Pb) in relation to cytopathologies in the gills and digestive gland of the mussel Mytilus edulis

Cell and tissue pathology of both, gill and digestive tissue, has been the subject of many studies for the elucidation of contaminant-induced biological effects. In the present study, cellular pathological alterations were linked to subcellular sites of chemical accumulation in gills and digestive gland tissues. For this purpose, mussels (Mytilus edulis) were exposed to the organic contaminants aroclor 1254 (PCB) (20 microg/L), phenanthrene (PAH) (150 microg/L) or the metal lead (Pb) (2.5mg/L). The localization of chemicals at the subcellular level was analysed by an antibody-based detection system (GSSP) by the use of commercially available antibodies specifically directed against the chemicals. Pathological changes were analysed in parallel in identical samples by transmission electron microscopy. After exposure to the different contaminants, cell organelles such as mitochondria, the endo-lysosomal system as well as endoplasmic reticulum showed clear evidence of chemically-induced alterations. Large numbers of crystalloid inclusions were found in mitochondria and in autophagic lysosomes as well as multi-lamellated whorls after PAH and aroclor exposure. Immunocytochemical detection of the chemicals showed their accumulation inside of various cell organelles such as lysosomes, mitochondria, and nuclei. Additionally, chemicals were localized in association to membranes, cilia and microvilli of gill and digestive gland cells. Furthermore, the chitinous rod and mucus secretions of gill epithelial cells were positively labelled for contaminants indicating their role in protection. Localization of contaminants by immuno-detection in combination with pathological diagnosis gives insights into the cellular targets of chemical attack.     

Stability of lysosomal and cell membranes in haemocytes of the common mussel (Mytilus edulis): effect of low temperatures.

Expanding industrial activities in the Arctic require an urgent assessment of the toxicity of chemicals at low temperatures. Organisms acclimatized to low temperature exhibit specific adaptations. For example, the amount of unsaturated lipids is increased to maintain the fluidity of the cell membranes. It has been hypothesized that such temperature-induced alterations in membrane lipid composition may affect the stability of lysosomal and cell membranes in the common mussel, Mytilus edulis, an organism exposed to seasonal temperature extremes. As mussels may be exposed to petroleum compounds along industrialized coastlines, we tested the combined effects of exposure to low temperature and the petroleum compound, phenanthrene, on haemocyte membrane stability. Test animals, acclimated to either 0 or 10 degrees C, were exposed to phenanthrene (0 = control or 500 micrograms l-1) and haemocytes were examined using the neutral red retention assay (lysosomal stability) and a fluorescence assay (cell membrane stability). At 0 degree C, lysosomal and cell membranes from uncontaminated mussels were destabilized compared with 10 degrees C (P = 0.0005). No significant effects (P > 0.05) of phenanthrene were detected at either temperature. Possible mechanisms underlying membrane destabilization include a weaker physical resistance of the membrane due to a higher amount of unsaturated lipids, a potentially higher level of reactive oxygen radicals at low temperature and the higher susceptibility of unsaturated lipids to oxidative stress. More work is required to better understand the consequences of this membrane destabilization at low temperature on the susceptibility of the organism to pollutants.     

Distribution of PACs in surficial sediments and bioavailability to mussels, Mytilus edulis of Halifax Harbour

Surficial sediments and blue mussels were collected around the perimeter of Halifax Harbour. Samples were analysed for the levels and fingerprint of polycyclic aromatic compounds (PACs). The objective was to explore the relationship between contaminant profiles in sediments and in mussels and to explore the possibility of deducing their sources, pattern of deposition and circulation in the harbour. Combustion derived compounds predominated in both sets of samples, with more variability observed in the level of petroleum derived hydrocarbons in mussel samples. Biota-sediment accumulation factors (BSAF, dry/dry wt.) varied throughout the harbour, ranging from means of 0.006 to 1.26, where generally fluoranthene displayed the highest BSAF. Sediments and mussels give complementary views of the geographical distribution of contaminants, they reflect the state of benthic and pelagic habitats. Results observed for Halifax Harbour are discussed from the perspective of published data.     

Effects of temperature on scope for growth and accumulation of Cd, Co, Cu and Pb by the marine bivalve Mytilus edulis

In many aquatic organisms including Mytilus edulis, the role of temperature on bioaccumulation of metals is still not clearly understood. In this study, uptake and accumulation of Cu, Co, Cd and Pb in mussels were investigated at different temperatures (6-26 degrees C). Results from exposure of isolated gills showed a positive relationship between temperature and metal uptake. But in whole organism experiments, only the accumulations of non-essential metals (Cd, Pb) showed a similar trend while the two essential metals Co and Cu were independent and inversely related to temperature, respectively. With exception of Cu, elimination process appeared to be independent of temperature. The study also showed that neither changes in scope for growth (SFG) of mussels nor chemical speciation could fully account for the observed temperature-effects. Overall, these results suggest that fundamentally (i.e. at epithelial membranes), temperature-effects on uptake are largely due to changes in solution chemistry and physical kinetics, which favours higher uptake at high temperature. But at whole organism level, complex physiological responses appears to mask the relationship, particularly for biologically essential metals like copper.