Cadmium accumulation in tissues of the mussel Mytilus galloprovincialis

Studies have been made on cadmium accumulation in tissues of mussels kept within 20-60 days in water artificially enriched by Cd up to 20-100 micrograms/l. Irrespectively of cadmium concentration in the medium, its accumulation in tissues decreases in the following order: mid-gut gland, gills, gonads, mantle, adductor. Maximum concentration of Cd was found in the digestive tubuli of the mid-gut gland by X-ray microanalysis. The increase in S and, to a lower extent, P concentrations in these tubuli was also observed. It is suggested that the latter is due to immobilization of Cd by metal-binding proteins as well as to lyzosomal vesicles involved into detoxication of Cd. The increase in the external cadmium up to 100 micrograms/l did not affect the level of K, Ca and Mg in tissues of the mussel.     

Cadmium accumulation in organs of the scallop Mizuhopecten yessoensis--II. Subcellular distribution of metals and metal-binding proteins

Cd accumulation in the hepatopancreas and gills of the scallop resulted in an increase of metal concentration in the cytoplasm, while in control animals the general amount of Cd in the hepatopancreas was concentrated in the microsomes and cytoplasm. Cd appears to be equally distributed among high molecular weight proteins and metallothioneins in control animals. It was shown that during the experiment the metal was bound mainly to high molecular weight proteins. After 30 days of exposure of scallops to flowing water Cd was redistributed to metallothionein (MT)-like proteins. Cd concentration in the lipids of the hepatopancreas of control and experimental scallops was equal.     

Prey selection and the functional response of sea stars (Asterias vulgaris Verrill) and rock crabs (Cancer irroratus Say) preying on juvenile sea scallops (Placopecten magellanicus (Gmelin)) and blue mussels (Mytilus edulis Linnaeus)

Predators in nature include an array of prey types in their diet, and often select certain types over others. We examined (i) prey selection by sea stars (Asterias vulgaris) and rock crabs (Cancer irroratus) when offered two prey types, juvenile sea scallops (Placopecten magellanicus) and blue mussels (Mytilus edulis), and (ii) the effect of prey density on predation, prey selection, and component behaviours. We quantified predation rates, behavioural components (proportion of time spent searching for prey, encounter probabilities) and various prey characteristics (shell strength, energy content per prey, handling time per prey) to identify mechanisms underlying predation patterns and to assess the contribution of active and passive prey selection to observed selection of prey. Sea stars strongly selected mussels over scallops, resulting from both active and passive selection. Active selection was associated with the probability of attack upon encounter; it was higher on mussels than on scallops. The probability of capture upon attack, associated with passive selection, was higher for mussels than for scallops, since mussels can not swim to escape predators. Sea stars consumed few scallops when mussels were present, and so did not have a functional response on scallops (the target prey). Rock crabs exhibited prey switching: they selected mussels when scallop density was very low, did not select a certain prey type when scallop density was intermediate, and selected scallops when scallop density was high relative to mussel density. The interplay between encounter rate (associated with passive selection) and probability of consumption upon capture (associated with both active and passive selection) explained observed selection by crabs. Scallops were encountered by crabs relatively more often and/or mussels less often than expected from random movements of animals at all scallop densities. However, the probability of consumption varied with scallop density: it was lower for scallops than mussels at low and intermediate scallop densities, but tended to be higher for scallops than mussels at high scallop densities. When mussels were absent, crabs did not have a functional response on scallops, but rather were at the plateau of the response. When mussels were present with scallops at relatively low density, crabs exhibited a type II functional response on scallops. Our results have implications for the provision of protective refuges for species of interest (i.e., scallops) released onto the sea bed, such as in population enhancement operations and bottom aquaculture.     

Isolation of a cadmium-releasing bacterium and characterization of its novel protease

Microorganisms were screened for their ability to release cadmium from scallop hepatopancreas, which is the main residue after removing of the edible parts of scallop. The isolated strain, 23-0-11, identified as Arthrobacter nicotinovorans, secreted a protease which released cadmium from scallop hepatopancreas into the liquid medium. The molecular mass of the enzyme was estimated to be 27 kDa. The sequence of the 15 N-terminal amino acids of the protease showed no close similarity with any other protein. Compared with a commercial enzyme, the purified protease had greater ability to release cadmium. The enzyme activity was greatest at 50 degrees C and pH 7.0, and was enhanced in the presence of Ca(2+), Mg(2+) and Mn(2+), while being strongly inhibited by Co(2+). The inhibition profile by the serine protease inhibitor, phenylmethylsulphonyl fluoride (PMSF), confirmed that the protease belonged to the serine protease family.     

Comparative evaluation of the toxicity induced by mussels (Mytilus galloprovincialis) from two sites of the Moroccan Atlantic coast in mice

Atlantic coast in mice. Preliminary studies showed that seawater contains heavy metals from domestic, agricultural and industrial wastes. Marine bivalves concentrate these pollutants by filtration and serve as vectors in human exposure. The objective of this study was to determine the concentration of heavy metals; cadmium (Cd); chromium (Cr), and lead (Pb) in mussels (Mytilus galloprovincialis) collected in two coastal sites; Jorf Lasfar (JL) (neighbouring a phosphate processing platform) and Oualidia (OL) (a vegetable growing area) located at 120 and 190 km south of Casablanca, respectively. Another objective was to test and compare the toxicity of these mussels on mice. The results indicated the presence of heavy metals (Cd, Cr, and Pb) in mussels at different concentrations, depending on the collection period. Higher concentrations were obtained at JL than at OL: for example, Cd concentrations were 80 +/- 15 to 199 +/- 28 versus 23 +/- 5 microg/g mussel dry weight, respectively. Cramming with mussel powder did not increase Cd, Cr, or Pb concentration in either liver or kidneys of treated mice. The relative kidney weights were reduced. Increased glucose urea was observed in animals' urine. Treatment with mussels from OL induced significant reduction (20%) in mice body weight, together with an increase in creatinuria. These results indicate that mussels collected from OL are more harmful than those obtained from JL are. All these mussels should not be recommended for human consumption.     

Superoxide dismutase activity in juvenile Litopenaeus vannamei and Nodipecten subnodosus exposed to the toxic dinoflagellate Prorocentrum lima

The toxic effect of the dinoflagellate Prorocentrum lima on juvenile American whiteleg shrimp Litopenaeus vannamei and giant lion-paw scallop Nodipecten subnodosus was evaluated. Organisms were exposed to three densities (500, 2000, or 5000 cells mL(-1)), superoxide dismutase activity and soluble protein in the hepatopancreas and muscle were determined at 1, 6, 24 and 48 h after challenge. Shrimp exposed at 5000 cells mL(-1) significantly increased SOD activity in the hepatopancreas at 1 h post-challenge, whereas enzymatic activity in muscle significantly increased at 24 h at all densities. Scallops exposed to 500 and 2000 cells mL(-1) showed significant SOD activity increase in hepatopancreas at 24 and 12 h, respectively. Mortality at 48 h was 100% in scallops exposed to 5000 cells mL(-1). Shrimp showed higher levels of SOD activity than scallops. Soluble protein content in the shrimp hepatopancreas was significantly higher at densities of 500 and 2000 cells mL(-1) at 6 and 1 h, respectively. Soluble protein content in the scallop hepatopancreas was higher than control values at 1 h after challenge. In this study, 500 cells mL(-1) was enough to trigger SOD activity in two benthic species exposed to the toxic dinoflagellate P. lima.     

Seasonal reproductive cycle in the mussel,Lamellidens corrianus

Seasonal reproductive cycle of the freshwater mussel, Lamellidens corrianus has been studied. These mussels are functional or simultaneous hermaphrodites. The spawning was at its peak during the months of September to December. The gonads were in growing stages with reduced gonadal activity during January to April, whereas the maturation of gonads was found to be intense during May to August.     

Bioaccumulation and Partitioning of Cadmium within the Freshwater Mussel Dreissena polymorpha Pallas

Kinetics of uptake, partitioning and elimination of cadmium were investigated in experimental studies with the freshwater mussel, Dreissena polymorpha. The radioactive cadmium isotopes ¹⁰⁹Cd and ¹¹⁵Cd were used as tracers. Shells, soft parts and body fluid of the mussel exhibited considerable differences in accumulation and elimination. Accumulation factors up to more than 70,000 were calculated for the periostracum, whereas accumulation factors for the whole mussels ranging up to 3,000 were calculated. The shells bound a great deal of cadmium, but only loosely, and the metal could be readily eliminated after transfer to uncontaminated water. In contrast, no significant amounts of the cadmium incorporated in the soft parts were eliminated. The results indicate that the major portion of cadmium in the soft parts is strongly bound and cannot be eliminated by exchange processes.     

Factors affecting trematode infection rates in freshwater mussels

Mussels are intermediate hosts of digenean trematodes, but determinants of these infections remain unknown. To address this problem, we collected duck mussels Anodonta anatina in eighteen lakes from northeastern Poland and examined how mussel age, sex, and the encrustation with zebra mussels Dreissena polymorpha and environmental conditions in lakes influenced infection rates. We also assessed parasitic preferences to host gonads and hepatopancreas and the impact of parasites on female fertility. Mussels were infected with Rhipidocotyle campanula and Phyllodistomum sp. Infection rates were higher in older and female mussels but were unrelated to the biomass of encrusting D. polymorpha and the trophy, thermal conditions, and Ca²⁺ availability. Parasites occupied gonads more often than hepatopancreas. Infected females were less likely to carry glochidia and incubated fewer glochidia. We suggest that the risk of infection by digenean trematodes increases with the amount of water processed by filter-feeding hosts and/or that parasites actively seek hosts which can provide them with abundant resources. This mechanism explains why parasites more often occupied older and female mussels and targeted their gonads. Future research on trematode-mussel interactions should integrate knowledge on different elements of the complex trematode life cycles, including effects of higher-order hosts such as fish.     

Tissue distribution of neutral deoxyribonuclease (DNase) activity in the mussel Mytilus galloprovincialis

The presence of neutral DNase activity in bivalves is reported for the first time. The enzyme activity in four tissues of the mussel Mytilus galloprovincialis was analyzed by three different methods (i) specific denaturating SDS-PAGE zymogram, (ii) sensitive single radial enzyme diffusion (SRED) assay and (iii) rapid and sensitive fluorimetric determination of DNase activity with PicoGreen. The fluorimetric assay was rapid and sensitive enough for determination of hydrolytic activity of dsDNA in mussel hepatopancreas, adductor, gills and mantle. Maximal activity in all mussel tissue extracts was obtained in the presence of Ca(2+) and Mg(2+) at pH 7.0 with dsDNA as substrate. The neutral DNase activity in mussel tissue decreases in order hepatopancreas, mantle>gills>adductor. The enzyme activity displays interindividual variability in particular tissue as well as variability among tissues within one specimen. In the hepatopancreas one to three distinct proteins expressing neutral, Ca(2+), Mg(2+)-dependent, DNase activity were detected by denaturating SDS-PAGE zymogram. This heterogeneity of neutral nucleases involved in DNA hydrolysis in hepatopancreas could reflect interindividual variability in mussel food utilization and nutrient requirement.     

An experimental study of the uptake and loss of Ra-226 by the tissue of the tropical freshwater musselVelesunio angasi (Sowerby) under varying Ca and Mg water concentrations

Freshwater musselsVelesunio angasi (Sowerby) from Magela Creek, Alligator Rivers Region, Northern Territory, Australia, experimentally exposed to mean elevated Ra-226 water concentrations ranging between 0.95 and 1.85 Bq 1^–1 for 28 days, accumulated Ra-226 in their tissue to mean concentrations ranging from 2.8 to 4.8 Bq per gram of dry tissue. The Ra-226 (log₁₀) was accumulated in a linear pattern over exposure periods of 28 and 56 days. Mussel size and sex had little or no effect on the rates of uptake of Ra-226 per gram of tissue.Increased Ca and Mg water concentrations, both in combination and singly, reduced the rate of uptake of Ra-226 by mussel tissue. The experimental data are consistent with Ra-226 accumulation being inversely proportional to both Ca and Mg water concentrations; for Ca the constant of proportionality i.e. is unity; for Mg it is about 0.1.The results indicate competitive inhibition of the uptake of Ra-226 by Ca, i.e. that the mussel treated Ra-226 as a metabolic analogue of Ca; however, there are other possible interpretations of these results that need not invoke competitive inhibition. For Mg the results suggest involvement of some other mechanism(s) apart from or in addition to competitive inhibition of Ra-226 by Mg.Exposure of mussels that had accumulated Ra-226 under field and laboratory conditions to radium-free water for up to 286 days resulted in no significant loss (P > 0.05) of Ra-226 from the tissue. This indicates a very long biological half-life for Ra-226 in the tissue ofV. angasi.     

Changes of Blue Mussels Mytilus edulis L. Lipid Composition Under Cadmium and Copper Toxic Effect

The lipid and fatty acid composition of the blue mussels Mytilus edulis L. gills and digestive glands was evaluated after 24 and 72 h of cadmium (Cd) and copper (Cu) exposure. Mussels were exposed to different cadmium (10, 100, and 500 μg/L) and copper (5, 50, and 250 μg/L) concentrations. Similar stress response of predominant membrane phospholipids level as well as polyenoic and non-methylene interrupted (NMI) fatty acids content was observed in mussel gills under both cadmium and copper effects. Increased NMI fatty acids level after 24 h, the metal ions treatment suggests that these acids contribute to the protective response to the membrane oxidative stress caused by accumulation of the metals. The content of cholesterol, some minor membrane phospholipids, and storage lipids (triacylglycerols, TAG) in the mussels’ organs alter significantly under the cadmium and copper effect. A two-step response at the digestive glands TAG level depends on the duration of the cadmium and copper treatments (24 and 72 h) on the mussels. The results demonstrate that Cd and Cu impact has adverse effects on gills and digestive glands lipid and fatty acids composition. The type of observed effects varies with the nature and concentration of the metal ions and depends on the role of the metals in the mussels’ life activity.     

Heavy Metal Accumulation and Distribution in Organs of the Mussel Crenomytilus grayanus from Upwelling Areas of the Sea of Okhotsk and the Sea of Japan

The subcellular and cytosolic distribution of Zn, Fe, Ni, Cu, Mn, Cd, and Pb in the digestive gland and kidney of the mussel Crenomytilus grayanus from upwelling areas of the Sea of Okhotsk and the Sea of Japan was studied. Cd, Zn, Pb, and Ni have accumulated in the kidneys of mussels from these areas. When the concentrations of both essential and toxic metals in the mussel organs had increased significantly, their redistribution into the cytosol took place. Gel chromatography of cytosolic proteins in the kidneys of mussels sampled in the area with a strong stationary upwelling revealed metallothioneins. This is uncommon for bivalve mollusks from unpolluted areas. High molecular weight proteins act as preliminary ligands for Cd.     

The effects of cadmium on ALA-D activity, growth and haemoglobin content in the water flea, Daphnia magna

The ALA-D activity, haemoglobin content and growth was studied in the water flea, Daphnia magna, exposed to 0, 0.1, 0.2, 0.4, 0.8 and 1.6 micrograms Cd/l. The ALA-D activity in water fleas exposed to 0.2-1.6 micrograms Cd/l fluctuated around the control value. The activity in animals exposed to 0.1 micrograms Cd/l decreased during the entire experiment. After 16 days exposure to cadmium the haemoglobin content in water fleas ranged between 80 and 31% of control value. In animals exposed to 0.8 and 1.6 micrograms Cd/l the haemoglobin content decreased progressively during the experiment. Growth was not affected by cadmium at these concentrations.     

镉胁迫对小报春幼苗生长及生理特性的影响

该研究采用盆栽控制试验,测定不同浓度(0、5、50、100、150、200 mg/kg)土壤Cd胁迫下小报春幼苗生长及生理生化指标,以探究小报春(Primula forbesii Franch.)对重金属镉(Cd)污染的抗性和敏感性,为新型香花地被植物应用于Cd污染土壤提供理论依据。结果表明:(1)低浓度(5 mg/kg)Cd胁迫能促进小报春株高和根长的伸长,高浓度(≥150 mg/kg)Cd胁迫下株高、根长和生物量明显降低。(2)随Cd胁迫浓度增加,小报春幼苗叶片光合作用、叶绿素含量、SOD和CAT活性先上升后下降,而其超氧阴离子产生速率、过氧化氢、丙二醛含量、叶相对电导率和POD活性持续升高。(3)在Cd胁迫条件下,小报春体内K、Zn含量降低,叶和根中Ca、Mg含量显著增加,体内Cd含量明显升高,且根系Cd含量远远高于叶和叶柄。研究发现,小报春幼苗对Cd胁迫具有一定的耐性,低浓度(5 mg/kg)Cd胁迫对小报春生长影响较小,但高浓度(≥150 mg/kg)Cd胁迫对小报春产生明显的毒害作用,影响其正常生长;小报春可能通过增强体内抗氧化酶系统活性、增加叶和根中Ca、Mg含量和根系截留镉来减轻镉胁迫的伤害,提高自身耐受镉胁迫能力。     

Impact of preparation method on gonad domoic acid levels in the scallop, Pecten maximus (L.)

The king scallop, Pecten maximus (L.), fishery is a valuable economic resource in the UK, and is reliant on supplying premium “roe-on” processed scallops to the continental market. A considerable degree of variability is observed in domoic acid (DA) levels among individual P. maximus and their body components, which complicates the management of the fishery during amnesic shellfish poisoning (ASP) events. This study examined the impact of professional processing and three differing laboratory preparation techniques on final gonadal DA levels. DA analysis was conducted using a LC–MS/MS procedure. The results demonstrate that different methods of preparation can significantly alter gonadal toxicities in scallops from the same site, and the extent to which DA within the digestive loop, which passes through the gonad, contributes to total gonadal DA. Mean gonadal toxicity attributed to the digestive loop contents was estimated at 4.7–24.7 μg DA g⁻¹. Despite large individual variations in toxin levels; in scallops with elevated gonadal toxicities resulting from higher digestive loop content toxicity, the effect of flushing out the contents of the digestive loop significantly reduced the DA content of the tissue and lowered the frequency of individuals harbouring levels above the 20 μg DA g⁻¹ statutory safety limit. Removal of the digestive loop contents can potentially result in an 87% decrease in gonadal DA burden. Furthermore, the method applied by professional processors effectively removed the contents of the digestive loop and reduced gonadal DA to levels comparable with the laboratory techniques. Deliberate contamination with scallop mucus did not increase gonadal DA levels. The extent of toxin variation resulting from differing gonad preparations demonstrates the need to standardize scallop tissue preparation techniques during ASP events. Consequently, detailed protocols aimed at minimizing the contamination of edible components should be developed and adhered to by both processing facilities and monitoring bodies.     

Comparative Analysis of Subcellular Distribution of Heavy Metals in Organs of the Bivalve Mollusks <Emphasis Type="Italic">Crenomytilus grayanus</Emphasis> and <Emphasis Type="Italic">Modiolus modiolus</Emphasis> in a Continuously Polluted Environment

The distribution of Zn, Fe, Ni, Cu, Mn, Cd, and Pb in subcellular fractions, and of Cd, Zn, and Cu in cytoplasm proteins of the kidney and digestive gland of the mussels Crenomytilus grayanus and Modiolus modiolus, sampled from contaminated and conditionally clean areas, was studied. It was found that, in a contaminated environment, the organs of mussels were more highly enriched with metals. It was shown that essential trace elements were accumulated mostly in the cytosol of organs of both molluscan species from contaminated areas, whereas in the background areas the trace elements were associated mostly with membrane structures in Gray's mussel, C. grayanus, and with the cytosol in M. modiolus, the northern horse mussel. The lead was bound mostly to membrane structures in organs of both mussel species at all stations. The method of gel chromatography enabled us to isolate metallothionein-like proteins from the kidney of the northern horse mussel sampled in contaminated areas, whereas their concentration in the kidney of Gray's mussels was lower than the limiting error of the method. It is supposed that in the kidney of Gray's mussel the synthesis of metallothionein-like proteins was quenched by the integrated effect of the accumulated metals.