Untreated Mussels Research Articles

Stress responses of bivalve mollusc Unio tumidus from two areas to ibuprofen, microplastic and their mixture.

Even though bivalve molluscs are recognized as bioindicators of freshwater quality, their responses to multiple stressors are unpredictable. This study aims to elucidate the inter-population peculiarities of the effect in the sub-chronic environmentally relevant exposure to novel contaminants. The specimens of Unio tumidus from reference (Pr) and contaminated (Ct) areas were treated with ibuprofen (IBU, 0.8 µg L-1), microplastic (MP, 1.0 mg L-1, size 0.1-0.5 mm), or their combination (Mix) for 14 days. Untreated mussels (PrC- and CtC-groups) served as controls. The PrC-group had higher levels of antioxidants Mn-SOD, Cu,Zn-SOD, catalase, and cholinesterase (AChE) as well as lesser levels of oxidative lesions (TBARS and protein carbonyls) in digestive glands, indicating lower environmental impact than in the CtC-group. However, lysosomal stability was similar in both control groups. Among antioxidants, Mn-SOD activity was affected most prominently, increasing in all exposed Ct-groups. TBARS level was increased only in PrMP-group compared to responsive control. IBU and Mix enhanced protein carbonyl concentration in the Pr-groups, and decreased it in the Ct-groups. AChE was induced in the CtIBU- and PrMix-groups, and lysosomal integrity increased in the CtIBU and CtMix-groups. Discriminant analyses indicated lesser differences between Pr-groups, demonstrating lower cumulative stress compared to Ct-groups. Generally, the most remarkable response was revealed in the CtIBU-group, and distortion of individual effects was established in combined exposures. The qualification of stress-neutral and stress-positive populations was proposed for Pr- and Ct-populations correspondingly. Inter-site peculiarities must be taken into consideration when the environmental impact of MP and pharmaceuticals is evaluated.

Electrical properties and aquatic ecotoxicity effects of ZnS nanocrystals

A new synthesis method of ZnS colloidal nanocrystals is presented in this work, and this novel approach allows one to produce Zinc sulfide nanocrystals owing to a modified polyol process that makes use of 1.3-propanediol as a solvent. The structure and morphology of the nanoparticles were characterized by high-resolution transmission electron microscopy, transmission electron microscopy, X-ray diffraction and spectroscopy photoluminescence. The results showed that quasi-spherical ZnS nanocrystals with sizes in the range 3–5 nm were formed. Their morphology, size and colloidal stability are driven by the polyol solvent. Under UV excitation, a strong visible PL emission, due to the radiative recombinations of the electron–hole pairs photo-generated in the ZnS nanoparticles, was observed. In order to study the environmental risk of ZnS, mussels Mytilus galloprovincialis were exposed to ZnS C1 $$=$$ 0.1 mg/L, ZnS C2 $$=$$ 1 mg/L and ZnS C3 $$=$$ 10 mg/L. Superoxide dismutase, catalase activities and protein thiols density were determined in the gill of treated and untreated mussels. No significant effects were detected in the gills of exposed mussels after 14 days of exposure compared to control. Optical property of the new nanomaterial, together with the easy synthesis process, makes the ZnS nanocrystals a promising candidate for applications optoelectronics and photocatalyst. The frequency-dependent impedance and AC conductivity have been investigated in this context. Impedance and modulus plane plots from 10 to $$10^{7}$$ Hz show the presence of bulk and grain boundary phases in ZnS at each measurement temperature from 125 to $$225\,^{\circ }\hbox {C}$$ . The activation energy was calculated from the temperature variation of DC conductivity. Studies of frequency and temperature dependence of electrical data of the compound suggest that conduction process in the material is thermally activated. Environmental risk assessment results constitute a scientific support for considering the use of this nonmaterial on various applications after further complementary studies in order to ensure the safety of these NPs.

Sex steroids and metabolic responses in mussels Mytilus galloprovincialis exposed to drospirenone

Drospirenone (DRO) is a synthetic progestin derived from 17α-spironolactone with a pharmacological mechanism of action similar to progesterone. Despite its wide use as pharmaceutical and consequent continuous release into the aquatic environment, DRO effects have been poorly investigated on aquatic biota. In order to unravel the toxicity mechanisms of DRO, mussels Mytilus galloprovincialis were exposed for 7 days to different concentrations of DRO, namely 20ng/L (Low; L), 200ng/L (Medium; M), 2000ng/L (High; H) and 10μg/L (Super High; SH) nominal doses. Following exposure, no significant effect was observed on gonad maturation of treated and untreated mussels. The levels of progesterone (P4) and testosterone (T) were measured in mantle/gonad tissues and no significant alteration detected after exposure. However, the application of a protonic nuclear magnetic resonance (1H NMR)-based metabolomics approach enabled a comprehensive assessment of DRO effects in mussels. Specifically, 1H NMR metabolic fingerprints of digestive glands of DRO treated mussel groups were clearly separated from each other and from controls through a principal component analysis (PCA). Moreover, a number of metabolites involved in different metabolic pathways were found to significantly change in DRO-exposed mussels compared to control, suggesting the occurrence of alterations in energy metabolism, amino acids metabolism, and glycerophospholipid metabolism. Overall, despite no changes in gonad maturation and steroids levels were recorded in mussels after DRO exposure, the metabolomics approach demonstrated its effectiveness and high sensitivity in elucidating DRO-induced metabolic disturbances in marine mussels, and thus its usefulness in the environmental risk assessment of pharmaceuticals.

Diversity of the molecular responses to separate wastewater effluents in freshwater mussels

The environmental safety of pharmaceutical and personal care products (PPCPs) requires a crucial examination. The aim of this study was to evaluate the responses of biomarkers of stress and toxicity in freshwater mussels to the effect of commonly found PPCPs in wastewater. We treated male mussels Unio tumidus, from an undisturbed site with ibuprofen (IBU, 250ngL−1), triclosan (TCS, 500ngL−1), or estrone (E1, 100ngL−1) for 14days. Untreated mussels from this site (C) and mussels inhabiting a polluted area (P) were also examined after a similar time of being kept in the laboratory. The consequences of chronic exposure of the mussels in the P-group were reflected in elevated concentrations of oxyradicals (1.4 times), oxidized glutathione (4.3 times), lipofuscin (2.2 times), and DNA-strand breaks in the digestive gland (DG) in comparison to the C-group, higher levels of caspase-3 activity in the DG, and vitellogenin-like proteins in gonads among all studied groups. Exposed mussels demonstrated some common responses with mussels in the P-group: elevated levels of lactate/pyruvate ratio, lipofuscin (IBU and E1), DNA fragmentation (TCS and E1), and caspase-3 activity (TCS and E1). Exposed to PPCPs mussels also showed elevation of ethoxyresorufin-O-deethylase and/or glutathione-S-transferase activity in the DG and a decrease in lysosomal stability in hemocytes (TCS and E1). The TCS group was distinguished by having the highest level of DNA-fragmentation and the lowest concentrations of total glutathione, oxyradicals, lipofuscin, pyruvate, and lactate, reflecting total metabolic depression. These results show that selected PPCPs at low concentrations alter a variety of physiological processes in this animal model system.

LATENT EFFECTS OF IRANIAN CRUDE OIL AND A CHEMICAL OIL DISPERSANT ON RED SEA MOLLUSCS

ABSTRACT Predation rate of the gastropod drill, Drupa granulata, on the mussel, Mytilus variabilis, was measured over a period of 28 days after adults from both species had been immersed for 168 h in seawater solutions containing high sublethal concentrations (10 ml/liter) of Iranian crude oil. Predation rate was three times higher in controls than in the group where both predator and prey had been exposed initially; intermediate values were determined among groups where only one species had been treated initially. Fecundity of drills, as evidenced by number of egg cases deposited, was directly related to mussel consumption. In a similar study with a chemical oil dispersant, exposure to high (0.003 ml/liter) sublethal levels for 168 h did not affect markedly the rate at which mussels were destroyed and consumed during post-treatment. However, the fecundity of untreated drills feeding on untreated mussels (controls) was 3 to 10 times greater than among groups in which one or both species had been exposed i...

Ionic regulation and shell mineralization in the bivalveAnodonta cygnea(swan mussel) following heavy-metal exposure

Freshwater mussels are one of the most imperiled faunistic groups in the world and environmental exposure to toxic heavy metals, which result in deregulation of calcium absorption and deposition in the laboratory, may be a contributing factor in their decline. To address potential effects of heavy-metal exposure on calcium transport and metabolism in freshwater bivalves, adult Anodonta cygnea (L., 1758) were exposed to a sublethal concentration (1.0 × 10−6 mol/L) of essential (Zn2+and Cu2+) or nonessential (Pb2+and Cr3+) metal for 30 days in the laboratory. Inorganic composition of extrapallial, haemolymph, heart, and pericardium fluids, and kidney tissue, as well as shell morphology by scanning electron microscopy, were compared in treated and untreated mussels. Calcium levels in fluids varied after exposure to any of the metals investigated, although the magnitude and threshold of effect were metal- and compartment-specific. Ca2+levels increased robustly in all fluids following exposure to Zn2+, Cu2+, or Cr3+, whereas levels decreased significantly in heart fluid alone following Pb2+exposure (p < 0.05). In constrast to exposure to the other metals, Cu2+revealed an interesting reverse-accumulation pattern, decreasing in the fluids but not in the kidney, where it clearly accumulates for excretion. In addition, whereas essential Cu2+and Zn2+are closely regulated, the nonessential metals Pb2+and Cr3+increase to very high levels. Drastic alterations in shell morphology, specifically the structure of border and inner pallial regions of the nacreous layer, were observed after Cu2+or Cr3+exposure. Collectively, data suggest that prolonged exposure to a sublethal concentration of these heavy metals can adversely affect compartmental calcium availability and shell composition in A. cygnea.

Heat shocking of Philippine green mussels, Perna viridis

SummaryThe use of different methods of heat shocking for green mussels was evaluated. These methods were heat treatment at 60 and 100 °C either in a hot water bath, in water‐saturated air or in dry heat. It was established that relaxation of the adductor muscles, which facilitates shucking, was quickest for the hot water bath treatment at both test temperatures. This treatment caused 100% of the mussels to be open at either 60 or 100 °C in 3–5 and 0.33–1 min exposure times respectively. The sensory acceptability of all the heat‐shocked mussel meats ranged from dislike slightly to like slightly. Generally, the increase in pH and water activity of the heat‐treated mussels, relative to the untreated mussels, was not statistically significant at the 5% level of significance.

Toxic vents and DNA damage: first evidence from a naturally contaminated deep-sea environment

Levels of DNA strand breakage were measured, using the comet assay, in cells from vent mussels, Bathymodiolus azoricus, from three contrasting vent fields on the mid Atlantic Ridge. Different levels of DNA damage were recorded in untreated mussels, shortly after collection, and it was animals from the shallowest, and less active, Menez Gwen vent field (840-m depth), which showed the greatest amount of damage. In contrast to animals from two deeper and putatively more toxic sites, Menez Gwen animals went on to repair this damage and were able to survive under laboratory conditions at 1 bar pressure for several months. Animals from the two deeper sites showed both higher levels of initial mortality and a much reduced capacity for survival at 1 bar. The differences in DNA damage levels at the time of collection were interpreted as an expression of differences in cell viability/enzyme activity rather than a reflection of any differences in their natural environmental conditions. Small B. azoricus showed a capacity to repair DNA damage, whereas this ability appeared to be lacking in large individuals. By reproducing at a relatively early age, the deep-sea vent fauna may be able to resist the toxic effects of its environment by exploiting this natural, stage specific capacity to repair damaged DNA.

Influence of abiotic factors on bacterial proliferation and anoxic survival of the sea mussel Mytilus edulis L.

The effect of several abiotic factors (salinity, temperature and pH) on bacterial proliferation and survival time of the sea mussel Mytilus edulis L. were studied under anoxic incubations. In addition, the presence in the incubation media of ammonium and the volatile fatty acids propionate and acetate, both excreted fermentation products of the bivalve, was tested. Anoxic incubations with seawater diluted with demineralised water showed at the lowest salinity (50% seawater, SW) a significant increase in the capacity of M. edulis to survive anoxia as compared to both 75% SW and control [100% SW, corresponding to 32 practical salinity units (psu)]. Formation of biotic sulphide and ammonium occurred in all incubations. However, bacterial proliferation was postponed by 2–3 days at lowest salinity and accordingly, concentrations of both compounds were lower. Anoxic survival profiles of mussels collected from different habitats in the Dutch Scheldt area, characterised by differences in salinity (range from 17 to 31 psu), corresponded with the above salinity effect. Walsoorden mussels (17 psu) showed the longest ( P<0.001) survival time under anoxia (LT 50=17.2 days) as compared with Paulina (27 psu) and Wemeldinge (31 psu) mussels (LT 50=12.8 and 9.8 days, respectively). Condition index (ratio of soft body weight to shell volume) was not correlated with anoxic survival time in untreated mussels, although this was clearly the case when the antibiotic chloramphenicol was added to the anoxic seawater. Acidification of the anoxic incubation medium had a positive effect on survival time. LT 50 values significantly ( P<0.001) increased from 10.2 days at pH 8.1 to 11.6 and 11.5 days at pH 7.3 and 6.5, respectively. Biotic sulphide and ammonium accumulation as well as bacterial numbers were significantly lower at pH 7.3 and 6.5 as compared with pH 8.1. Anoxic incubations at 10 °C (LT 50=12.0 days) strongly increased survival time as compared to 18 °C (LT 50=5.9 days). The benefit of antibiotic addition was also stronger at lower temperature (10 °C). Addition of both propionate and acetate (0.5 mM) displayed no effect on mortality of mussels under anoxia, but ammonium (0.5 mM) caused a negative effect ( P<0.001). Biotic sulphide and ammonium concentrations measured in both volatile fatty acid incubations were lower than the control situation, as well as total bacterial numbers. This study shows that environmental factors play a significant role in determining the course of bacterial infection and death of bivalves exposed to anoxia.

Cytochemical Responses in the Digestive Tissue of Mytilus edulis Complex Exposed to Microencapsulated PAHs or PCBs

Cytochemical responses in the digestive tissue of Mytilus edulis complex fed a microencapsulated mixture of PAHs (composed of phenanthrene, fluoranthene and benzo[a]pyrene) or PCBs (Aroclor 1254) were evaluated. Lysosomal membrane labilization period and lipofuscin content of digestive tissue were significantly decreased and increased, respectively, after 30 days of exposure to PAHs but not PCBs. NADPH-ferrihemoprotein reductase activity in the digestive tissue was significantly increased after 30 days exposure to both PAHs and PCBs, whereas catalase activity was significantly increased only after exposure to PCBs. Neutral lipid content and gamma-glutamyl transpeptidase activity of digestive tissue of mussels were significantly increased and decreased, respectively, after 30 days exposure to both PAHs and PCBs when compared to untreated mussels, but were not significantly different when compared to mussels treated with com oil representing the vehicle control. Exposure to PAHs induced peroxisome proliferation in the digestive tissue of M. edulis complex. The observed cytochemical responses in the digestive tissue of M. edulis complex exposed to PAHs and PCBs in the laboratory are comparable to the cytochemical responses observed in the digestive tissue of mussels collected in the field from sites contaminated with predominantly PAHs and PCBs.

Innate Immunity: ISOLATION OF SEVERAL CYSTEINE-RICH ANTIMICROBIAL PEPTIDES FROM THE BLOOD OF A MOLLUSC, MYTILUS EDULIS

We have isolated from the blood of immune-challenged and untreated mussels (Mytilus edulis) antibacterial and antifungal peptides. We have characterized two isoforms of a novel 34-residue, cysteine-rich, peptide with potent bactericidal activity and partially characterized a novel 6.2-kDa antifungal peptide containing 12 cysteines. We report the presence of two members of the insect defensin family of antibacterial peptides and provide a phylogenetic analysis that indicates that mollusc and arthropod defensins have a common ancestry. Our data argue that circulating antimicrobial peptides represent an ancient host defense mechanism that predated the separation between molluscs and arthropods at the root of the Cambrian, about 545 million years ago.

Free-radicals: Qualitative evaluation at bedside

Studies were performed on the common mussel, M. edulis L., to determine whether copper (Cu) exposure can affect the extent to which digestive cell proteins are oxidised and whether such oxidative damage is mediated by free radicals. Three age groups of mussels were exposed for 6 -days to environmentally realistic concentrations of Cu and then digestive gland homogenates were examined for evidence of protein carbonyl formation. Significant increases in carbonyls relative to untreated control mussels were seen for the youngest (2–4 year-old) and oldest (≥ 10 year-old) mussels only after exposure for 6 days, followed by recovery from exposure for a further 6 days. Untreated mussels also showed an age-related difference in protein oxidation, with a significantly lower concentration in the youngest animals (2–4 year olds). Copper did not affect the levels of modified tryptophan or tyrosine residues or the extent of total lipid peroxidation in digestive gland homogenate. Significant depletion of total vitamin E (a-tocopherol) was seen only in young and medium-aged mussels following exposure for 6 days. The levels of protein carbonyl groups were increased in digestive cell cytosol and lighter lysosomes but not in heavier lysosomes or digestive gland microsomes following 5 days exposure to Cu. Dihydrohodamine-123 was converted to fluorescent rhodamine-123 following sequestration into digestive cell lysosomes. The results suggest a link between the lysosomal sequestration of copper, a concomitant increase in the production of oxyradicals and the potential for intracellular oxidative damage, as well as an increased capacity for oxidative damage in older animals.

The role of oxyradicals in intracellular proteolysis and toxicity in mussels

Studies were performed on the common mussel, M. edulis L., to determine whether copper (Cu) exposure can affect the extent to which digestive cell proteins are oxidised and whether such oxidative damage is mediated by free radicals. Three age groups of mussels were exposed for 6 -days to environmentally realistic concentrations of Cu and then digestive gland homogenates were examined for evidence of protein carbonyl formation. Significant increases in carbonyls relative to untreated control mussels were seen for the youngest (2–4 year-old) and oldest (≥ 10 year-old) mussels only after exposure for 6 days, followed by recovery from exposure for a further 6 days. Untreated mussels also showed an age-related difference in protein oxidation, with a significantly lower concentration in the youngest animals (2–4 year olds). Copper did not affect the levels of modified tryptophan or tyrosine residues or the extent of total lipid peroxidation in digestive gland homogenate. Significant depletion of total vitamin E (a-tocopherol) was seen only in young and medium-aged mussels following exposure for 6 days. The levels of protein carbonyl groups were increased in digestive cell cytosol and lighter lysosomes but not in heavier lysosomes or digestive gland microsomes following 5 days exposure to Cu. Dihydrohodamine-123 was converted to fluorescent rhodamine-123 following sequestration into digestive cell lysosomes. The results suggest a link between the lysosomal sequestration of copper, a concomitant increase in the production of oxyradicals and the potential for intracellular oxidative damage, as well as an increased capacity for oxidative damage in older animals.

Arylsulphatase activity associated with phenanthrene induced digestive cell deletion in the marine mussel Mytilus edulis.

The acid hydrolase arylsulphatase has been localized at the ultrastructural level in digestive cells of the marine mussel Mytilus edulis for control and phenanthrene-treated (200 micrograms/l) animals. In untreated mussels the activity was generally restricted to the lysosomal-vacuolar system and the Golgi apparatus. It was associated with all types of vesicle, although not all individual vesicles were reactive. In heterolysosomes which were filled with precipitate the reaction product was most densely associated with the limiting membranes. Lipid inclusions commonly occurred in the digestive cells; these sometimes showed limited reaction for enzyme activity. The striking difference between normal and phenanthrene-treated samples was the presence in all treated animals of reaction product in the inter-cellular spaces and varying degrees of cytoplasmic activity in a number of digestive cells. This is interpreted as a sign of impending cell deletion. Sections for morphological examination showed evidence of increased digestive cell deletion in phenanthrene-treated mussels. The process results in release of membrane-bound bodies into the tubule lumen.

Distribution of cadmium, zinc and copper in the musselMytilus edulis. Existence of cadmium-binding proteins similar to metallothioneins

The study of the distribution of Cd, Zn and Cu in homogenates of mussels has shown that Zn and Cu are principally associated with high mol. wt. proteins. The same distribution is observed for Cd in untreated mussels, but in Cd chronically intoxicated animals, the metal is principally bound to low mol. wt. proteins synthesized in response to the uptake of the cation and similar to metallothioneins of vertebrates.