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PubMed1981

Monoamine precursor uptake in ganglia of a mollusc Anodonta cygnea L. (Bivalvia) An autoradiographic and ultrastructural study.

In vitro-uptake and localization of 3H-5 hydroxytryptophan and 3H-dihydroxyphenylalanine have been investigated by means of light microscopic autoradiography in the central nervous system of the freshwater mussel, Anodonta cygnea L. Accumulation of the labelled monoamine precursors could be observed both over neuronal perikarya and neuropil in the ganglia. 3H-5-hydroxytryptophan and 3H-dihydroxyphenylalanine were taken up by neurons characterized by a small size (10-20 micrometer), a polygonal form and an intense staining with toluidine blue. Labelled axon branches situated either in the neuropil or between nerve cells in the cortex exhibited only a moderate activity, frequently of a diffuse character. After pretreatment fro 24 h with colchicine, silver grains were also seen over a neuronal perikarya and some axon profiles. This shows that local uptake of the monoamine precursors and, presumably, the synthesis of the monoamines takes place both at perikaryal and axonal/terminal level. The nerve cells accumulating the monoamine precursors contain a nucleus with rich chromatin content and a deeply invaginate nuclear envelope. Further ultrastructural characteristics of these neurons are their well-developed rough endoplasmic reticulum system, a great number of mitochondria and glycogen granules, and the presence of dense-cored vesicles of different types. All those features are responsible for the electron-dense appearance of the cytoplasm.

5-Hydroxytryptophan↗
PubMed2001

Accumulative characteristics of pesticide residues in organs of bivalves (Anodonta woodiana and Corbicula leana) under natural conditions.

Accumulative characteristics of pesticide residues in the gill, midgut gland, gonad, and the remaining tissues of the bivalve mollusks Anodonta woodiana and Corbicula leana were examined during the rice planting seasons of 1992 and 1994. Although seven pesticides, except thiobencarb, were accumulated all at ppb levels in the midgut gland (liver) and gonad of both bivalves during their application period, thiobencarb was accumulated in C. leana at extremely high levels of 15.70 microg g(-1) in 1992 and 12.45 microg g(-1) in 1994 in the midgut gland and 15.80 microg g(-1) in 1992 and 16.40 microg g(-1) in 1994 in the gonad, respectively. These levels were about 100 times higher than those in A. woodiana. Thiobencarb and molinate in A. woodiana and chlornitrofen (CNP) and molinate in C. leana remained in the gonad and midgut gland longer than in the gill and remaining tissues, while thiobencarb in organs of C. leana remained at ppm levels until the end of the experiments. The present study suggests that these interspecies differences can be attributed to differences between the two species in their ability to eliminate pesticides.

Animals↗
PubMed1989

Distinct glutathione-dependent enzyme activities and a verapamil-sensitive binding of xenobiotics in a fresh-water mussel Anodonta cygnea.

A fresh-water mussel Anodonta cygnea, an aquatic invertebrate resistant to pollution, possesses an inherent high potential to bind 2-acetylaminofluorene onto membrane vesicles. This binding is saturable and trypsin- and verapamil-sensitive. Simultaneously, this mussel reveals a relatively high inherent activity of glutathione-dependent enzyme activities with a distinct spectrum of substrate affinities. Both these activities are similar to the elements of the molecular mechanism involved in the acquired multi-drug resistance phenomenon described in tumor cell-lines. The recognition that in organisms exposed to polluted waters a multi-xenobiotic resistance mechanism may be involved is essential for understanding both the biological impact of pollution and the development of methods for rational risk assessment in regulatory policy.

2-Acetylaminofluorene↗
PubMed1991

How is O2 consumption maintained independent of ambient oxygen in mussel Anodonta cygnea?

The mechanisms of adaptation allowing resting freshwater mussels Anodonta cygnea to maintain their oxygen consumption (MO2) constant when the O2 partial pressure in the inspired water (PIO2) varied were studied at 13 degrees C. Steady-state values of oxygen consumption and/or shell valve activity were determined at prefixed PIO2 for periods ranging from 2 to 15 days. Values of PO2, O2 concentration and acid-base status of arterial blood in the heart were determined after two days. MO2 was maintained constant over PIO2 ranging from 35 to less than 1 kPa. At 0.3 kPa it decreased by 50%. Valves remained open (and MO2 constant) most of the time even during a 4.5 day period at PIO2 approximately 1.5 kPa. Between 35 and 1 kPa, blood PO2 at the heart level can remain low and within a narrow range independent of PIO2. Blood PCO2 increased at high PIO2 and decreased at low PIO2. Data are compared to previous results in crayfish and in wels (sheat-fish). On the basis of data similarity it is proposed that these three animals exhibit the same basic strategy for maintaining resting MO2 when PIO2 varies. This common feature relies mainly on the ability to maintain PO2 in the arterial blood at a value which is low and independent of PIO2. Hence, in terms of O2, homeostasis of the milieu intérieur is accomplished.

Acid-Base Equilibrium↗
PubMed1996

Accumulation of extractable organic halogens (EOX) by the freshwater mussel, Anodonta cygnea L., exposed to chlorine bleached pulp and paper mill effluents.

Freshwater mussels, Anodonta cygnea L., were collected in a lowly polluted area in north-eastern France, and exposed during 28 days at three sites up and downstream of a chlorine bleached pulp and paper mill effluent Water, sediment and mussel samples were collected at regular intervals and analyzed for halogenated organic residues using the sum parameters AOX (Adsorbable Organic Halogen) and EOX (Extractable Organic Halogen). The survey of the water showed a chronic contamination by organic halogens of the water and the sediment downstream of the pulp and paper mill sewer. Mussels exposed upstream of the mill showed no significant variation of their EOX tissue concentration. In contrast, progressive accumulation of EOX occurred with the exposure time in mussels exposed downstream of the mill. No steady state of the EOX concentration was observed after four weeks of exposure.

Animals↗
PubMed1990

Uptake and body distribution of chlorinated phenolics in the freshwater mussel, Anodonta anatina L.

Freshwater mussels (Anodonta anatina L.) were exposed to [14C]pentachlorophenol (PCP) and [14C]3,4,5-trichloroguaiacol (CG-3) under laboratory conditions. Uptake and body distribution in mussels as well as total water-soluble metabolites of chlorophenolics in hemolymph and digestive gland were measured. The time course of chlorophenolic accumulation in the mussel soft tissue was followed by analyzing the decrease in the radioactivity in exposure water. The bioconcentration factors (BCFs, activity in animal per activity in water) were measured at steady state for the soft tissue homogenate and separate organs. Both chlorinated phenolics reached a steady-state concentration during the first 24 hr. BCFs in soft tissue ranged from 145 to 342 for PCP and 34 to 125 for CG-3. Accumulations by the digestive gland (hepatopancreas) and kidneys were 2 and 1.3 times greater, respectively, than the average accumulation by the whole soft tissue. The water-soluble fraction of PCP (1-8%) and CG-3 (0.4-2.9%) in separate organs implied only a minor metabolism of chlorophenolics in this animal.

Animals↗
PubMed1991

Accumulation and depuration of chlorinated phenolics in the freshwater mussel (Anodonta anatina L.).

Uptake from ambient water and the depuration of five chlorinated phenolics, two chloroguaiacols (3,4,5-tri- and tetrachloroguaiacol), and three chlorophenols (2,4,6-tri-, 2,3,4,6-tetra-, and pentachlorophenol) were studied in the duck mussel (Anodonta anatina). Groups of animals were exposed at four acclimiation temperatures (3, 8, 13, 18 degrees C) to four chlorophenolic concentrations (total 6-56 micrograms/liter). The depuration was monitored for 72 hr. For the analysis of individual chlorophenolics by the GC/ECD technique, the soft tissue of mussels was homogenized, spiked with internal standard, acetylated, and extracted with n-hexane. The bioconcentration factors (BCF) (concn. in animal wet wt./concn. in water) were determined for mussel soft tissue. The highest BCF was found for pentachlorophenol (81-461) and the lowest for trichlorophenol (14-125). Neither water temperature nor exposure concentration affected the BCFs. The compounds studied were depurated rapidly and their depuration half-lives (T1/2) in soft tissue were generally less than 24 hr.

Animals↗
PubMed1990

The micronucleus assay in Anodonta cygnea for the detection of drinking water mutagenicity.

A micronucleus test in gill cells of the freshwater mussel Anodonta cygnea has been proposed for the detection of drinking water genotoxicity. Animals were exposed for 28 days to a drinking water sample and collected every week. Highly significant increases in spontaneous MN frequency were observed at each sampling, especially after 13 days of exposure. As positive control 2 doses of mytomicin C (MMC) were used (10(-8) and 10(-7) M). A second experiment was performed at a municipal waterworks in order to assess the role of water treatment processes in the production of mutagenic compounds. The most prevalent genotoxic effects were detected after chlorination (mean: 10.47% +/- 3.05, p less than 0.001).

Animals↗
PubMed1979

ATPase stimulated by Na+ or K+ in gills of the freshwater mussel Anodonta.

1. Microsomal preparations from the gills of the freshwater mussel anodonta cygnea cellensis show Mg2+ -dependent Na+ - or K+ -stimulated ATPase activity, which is not inhibited by ouabain. 2. Na+ - or Ka+ -ATPase activity is decreased by Ca2+, acetylcholine, choline, and tetramethylammonium, but slightly increased by ethyl alcohol. 3. It is tentatively suggested that Na+ - or K+ -ATPase is involved in the mechanism of active monovalent cation uptake through the gills of freshwater mussels.

Adenosine Triphosphatases↗
PubMed1982

Subcellular localization and characterization of HCO3(-)-ATPase from the mantle of the freshwater clam, Anodonta cataracta.

1. HCO3(-)-stimulated ATPase activity was demonstrated in mantle tissue of the freshwater clam, Anodonta cataracta. 2. Calcium (1 mM) slightly inhibited and SCN- completely inhibited HCO3(-)-stimulation of the enzyme. 3. ATPase activity had a Km of 6.8 mM for HCO3(-)-activation and was inhibited at HCO3(-)-concentrations greater than 20 mM. 4. Subcellular fractionation studies revealed the presence of both a mitochondrial and a non-mitochondrial HCO3(-)-ATPase.

Adenosine Triphosphatases↗
PubMed1984

Succinate dehydrogenase in various tissues of Anodonta couperiana, Elliptio buckleyi and Mercenaria campechiensis (Mollusca: Bivalvia).

The specific activities, kinetic constants and pH optima for succinate dehydrogenase were determined in mantle, gill and adductor muscle from three bivalves (Anodonta couperiana, Elliptio buckleyi and Mercenaria campechiensis). In general the Km values for succinate and Ki values for fumarate differed in all bivalve tissues whereas the Ki for malonate was consistently low. The ratio of Ki (fumarate) to Km (succinate) was less than one for all tissues studied. This is similar to many facultative anaerobes with fumarate reductase activity.

Animals↗
PubMed1987

Interactions of organic pollutants with gills of the bivalve molluscs Anodonta californiensis and Mytilus californianus: uptake and effect on membrane fluxes. II.

1. The uptakes of 2,4,5-T, glyphosate, parathion, paranitrophenol, naphthalene, glycine, and inulin by gills of the bivalve molluscs Anodonta californiensis (freshwater) and Mytilus californianus (marine) show non-polar compounds are taken up to a greater extent than polar compounds except where active transport occurs. 2. The uptake of glycine by M. californianus is reduced by pollutants containing complexing functional groups but not by non-polar compounds. 3. The uptake of parathion alters the polyphosphate-inorganic phosphate balance in M. californianus. 4. The uptakes of pollutants parallel their toxicities toward rats.

Animals↗
PubMed2004

Accumulation and depuration of cyanobacterial paralytic shellfish toxins by the freshwater mussel Anodonta cygnea.

The increasing frequency by which the production of paralytic shellfish toxins (PST) by freshwater bloom-forming cyanobacteria is being noticed world-wide raises the possibility of PST bioaccumulation by freshwater mussels. This study evaluates PST accumulation and depuration by the freshwater mussel Anodonta cygnea exposed over a 14-day period to high densities (mean = 1.4 x 10(9) cells1(-1), S.D. = 0.29 x 10(9) cellsl(-1)) of the toxic cyanobacterium Aphanizomenon issatschenkoi (corresponding to a mean toxin concentration of 25.5 nmol PSTl(-1), S.D. = 9.9 nmol PSTl(-1)). Mussels were subsequently detoxified either by starvation or by feeding on the non-toxic green-algae Ankistodesmus falcatus. Filter feeding activity and toxin uptake by the mussels were followed by cell counting and toxin analysis in water samples taken before and after each daily water renewal. The accumulation and depuration of PST as well as the anatomical distribution of toxins were monitored throughout the experiment by HPLC analysis of mussel extracts. Mussels fed the toxic cyanobacterium removed on average 65.3% of cells and 40.36% of total PST daily provided. Daily rates of cell clearance (% of initial) were negatively correlated with the amounts of PST daily provided (but not with the amount of cells). This suggests a negative effect of toxins on the feeding behaviour of mussels. Small amounts of toxins could be detected in the mussels after the second day of exposure, reaching a maximum of 26 microg PST100 g(-1) by day 7. The viscera contained the greatest proportion of toxins (78%) at the start of the toxification. However, increasing amounts of PST were found in the remaining tissues (gills, mantle and foot) over time. Toxins detected in the mussel extracts were the same provided in the dietary A. issatschenkoi. Nevertheless, mussels showed a higher proportion of saxitoxin and decarbomoylsaxitoxin and a lower proportion of gonyautoxin-5 than the fed cyanobacterium. Similar depuration efficiencies were observed among starved individuals (6.9% day(-1)) and those fed with A. falcatus (8.2% day(-1)) indicating that both treatments had comparable effects on toxin metabolism. Mussels showed a typical S shaped depuration kinetics curve consisting of a first short period of slow toxin decay followed by a rapid loss and a subsequent slower release of toxins. Trace to undetectable levels of PST were found in mussels after the 14-day depurating period. Although freshwater mussels are not widely consumed by humans, their capacity to accumulate PST points to the risk of PST propagation through the food chain of freshwater ecosystems via filter-feeding mussels.

Analysis of Variance↗
PubMed2004

H+-ATPase of crude homogenate of the outer mantle epithelium of Anodonta cygnea.

The outer mantle epithelium of the freshwater bivalve, Anodonta cygnea, is responsible for the mineralisation of the shell. Under short circuit conditions, it generates a current that is due to the operation of an electrogenic proton pump located in the apical barrier of that epithelium. Bafilomycin A1 and Concanamycin A inhibited the short circuit current. The IC50 and maximum inhibition dose were 0.17 and 0.5 microM for Bafilomycin A1, and 0.7 and 5 microM for Concanamycin A. The present work was undertaken to further characterise V-type ATPase of the outer mantle cells. The V-ATPase enzymatic activity of crude homogenate, measured as the amount of inorganic phosphorous released, due to ATP hydrolysis, in the presence of Na2SO3 (200 mM) was found to be 4.6+/-1.1 micromol Pi/mg protein/h, at 27 degrees C, pH 7.0-7.4 and ATP 4.5-6.0 microM. Bafilomycin A1 and Concanamycin A inhibit the V-ATPase activity with an IC50 of 14 and 8 nmol mg(-1), respectively. Dicyclohexylcarbodiimide (DCCD; 100 mM) and NaNO3 (100 microM) inhibited the V-type ATPase in what it seems a non-specific manner and NEM (100 mM) was unable to do it. Bafilomycin A1 (10 microM) and Concanamycin A (10 microM), inhibited 50-60% of the total activity.

Adenosine Triphosphate↗
PubMed2005

Isolation, purification and characterization of glycosaminoglycans in the fluids of the mollusc Anodonta cygnea.

The isolation, purification and characterization of Anodonta cygnea haemolymph, and extrapallial fluid glycosaminoglycans (Gags), which have high calcium affinity, were carried out in order to better understand the process of nacreous shell biomineralization. Our results show the existence of two different Gags with similar contents in the two fluids, throughout the year, but with significant seasonal variations for both. In the winter months, we identified by electrophoresis, only one kind of Gag chain (hyaluronic acid) while two different Gags (hyaluronic acid and heparan sulphate-like) from both fluids of A. cygnea were found in summer months. Quantification showed the total Gag fractions of both fluids in average, at their highest concentration (79.8 mg/L) in the highest calcification summer period. In contrast, the period of the year with the lowest concentration of total Gags (54.6 mg/L) occurred during the winter when calcification is reduced. This significant decrease between seasons is correlated mainly with the sulphated fraction, being 37.1 mg/L in the summer while only 9.2 mg/L in the winter haemolymph. The present data suggest that a heparan sulphate-like Gag has a relevant role in the biomineralization mechanisms acting as the calcium carbonate nucleator in the shell.

Animals↗
PubMed2002

Assessment of bioavailability and effects of chemicals due to remediation actions with caging mussels (Anodonta anatina) at a creosote-contaminated lake sediment site.

A study was conducted at Lake Jämsänvesi in Central Finland, to identify the potential ecotoxicological risks of the remediation operation of a creosote-/PAH-contaminated lake sediment, made by capping during the years 1998-1999. Mussels (Anodonta anatina) were deployed to the lake at the same time as the remediation operation was started in November 1998. The contaminated area (0.5 ha) was covered by a filter geotextile (polypropylene), gravel and sand (1-1.5m) which were spread out on the ice and let to sink onto the bottom of the lake when the ice melted in May 1999. The possible impacts of capping to the adjacent environment were assessed from mussels exposed and particulate material settled (SPM) to collectors placed on the lake bottom. Mussel tissue, SPM, the water inside the collector were analyzed for polycyclic aromatic compounds (PAH)-compounds. Biological endpoints included body condition, glycogen and protein contents of adductor muscle. Mussels and SPM exposed downstream to the contaminated site (Site 3) contained the highest total PAH concentrations. Biota-sediment accumulation factors of acenaphthene, phenanthrene, anthracene, fluoranthene, pyrene and benzo(a)anthracene of mussels varied from 0.79 to 1.45. The glycogen and protein concentrations were lowest in adductor muscle from mussels exposed to conditions at Site 3. Concentrations of some PAH-compounds were found distinctly increased adjacent to the remediated area, possibly due to the agitation of contaminated sediment due to the capping. It is also possible that resuspension of sediment around remediated area (containing some PAHs) spread the deposited PAH-compounds.

Animals↗
PubMed2001

Avoidance responses to aluminium in the freshwater bivalve Anodonta cygnea.

This study examined the effect of aluminium (Al) on the filtering behaviour (shell opening or gape) of the freshwater bivalve Anodonta cygnea L in neutral fresh water. Parallel measurements of Al concentration in the soft tissues were made to examine the relationship between changes in behaviour and accumulation of Al. The number of lysosomal granules in the gill, kidney and digestive gland were counted, as lysosomes are known to be involved in the excretion and detoxification of trace metals. The bivalves were exposed to two environmentally relevant concentrations of added Al i.e. 250 and 500 microg l(-1) (9.25 and 18.5 microM l(-1)) at neutral pH for 15 days and shell movement monitored continuously. Aluminium affected the mussels' filtering activity, producing an avoidance reaction whose magnitude was concentration-dependent; 250 microg l(-1) added Al produced no detectable change, while 500 microg l(-1) Al reduced mean duration of shell opening by 50%. This effect was irreversible over a 15 day recovery period. Tissue levels of Al after 15 days exposure were an order of magnitude higher in animals exposed to 250 microg l(-1) added Al than in those exposed to 500 microg l(-1). This was consistent with the inhibition of filtering activity due to valve closure at the higher concentration, which may have prevented uptake of Al. In addition, probable different chemical speciation of Al in the water column (soluble for 250 and colloidal for 500 microg l(-1)) may lead to marked differences in tissue uptake. The kidney and digestive gland were the main sites of accumulation of Al and concentrations remained significantly elevated 15 days after transfer of animals to clean water. It is suggested that mucus plays a role in the exclusion of Al as elevated concentrations were measured in the pseudofaeces of animals during and after exposure. Lysosomal granules may be involved in the intracellular handling and detoxification of Al as numbers increased significantly in all organs during exposure and continued to increase after the animals were transferred to clean water. The present study provides evidence for the bioavailability and toxicity of Al to mussels at neutral pH and at concentrations which are known to enter neutral freshwaters when mobilised by natural or anthropogenic acidity. The changes in behaviour and uptake of Al in the mussel observed in this investigation are, therefore, likely to be reflected in the natural environment and the degree to which Al affects the 'fitness' of the mussel populations and the transfer of Al through the food chain merit investigation.

Aluminum↗