Freshwater Bivalve Anodonta Cygnea Research Articles

Effect of sodium chloride on oxidative stress biomarkers of the freshwater bivalve Anodonta cygnea

For the first time a study was conducted of the effects of the change in the concentration of sodium chloride in water on the biomarkers of oxidative stress (enzymes: catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST), and the levels of reduced glutathione (GSH), and a marker of peroxidation lipids (LPO) – malonic dialdehyde (MDA)) in the gills of the freshwater mollusc Anodonta cygnea. Adult specimens of A. cygnea of both sexes with a shell length of 70–100 mm were selected for the study, a total of 36 individuals. The molluscs were collected in one of the bays on the southwestern coast of the Volga reaches of the Rybinsk Reservoir (reservoir of the cascade of the upper Volga). The molluscs were kept in the laboratory in aquariums with river water and sand collected from the place of the molluscs' collection. During the experiment, a solution of NaCl 3 g/l in river water was introduced into the container with molluscs. Samples of molluscs’ tissues were taken before application of the saline solution, 40 and 120 min after changing the salt concentration to 3 g/l, and 20 and 60 min after changing the NaCl solution to river water. The change in the concentration of NaCl in water (0–3 g/l) did not affect the content of water-soluble protein in the gills of molluscs during short-term exposure. The change in NaCl concentration in water caused the intensification of LPO processes in the gills of A. cygnea. The change in the activity of the enzymes of the antioxidant system and the increase in the concentration of GSH in A. cygnea gills, indicates their involvement in protecting freshwater molluscs from NaCl-induced oxidative stress. The change in the functioning of the glutathione system may be a compensatory mechanism for changing the NaCl concentration in water. Such biomarkers of oxidative stress as malonic dialdehyde and enzymes of the antioxidant system are sensitive indicators of changes in NaCl concentration in water in freshwater bivalve molluscs, for example A. cygnea.

Biomineralization Studies on Cellulose Membrane Exposed to Biological Fluids of Anodonta cygnea

The present work proposes to analyse the results obtained under in vitro conditions where cellulose artificial membranes were incubated with biological fluids from the freshwater bivalve Anodonta cygnea. The membranes were mounted between two half 'Ussing chambers' with different composition solutions in order to simulate epithelial surfaces separating organic fluid compartments. The membrane surfaces were submitted to two synthetic calcium and phosphate solutions on opposite sides, at pH 6.0, 7.0 or 9.0 during a period of 6 hours. Additional assays were accomplished mixing these solutions with haemolymph or extrapallial fluid from A. cygnea, only on the calcium side. A selective ion movement, mainly dependent on the membrane pore size and/or cationic affinity, occurred with higher permeability for calcium ions to the opposite phosphate chamber supported by calcium diffusion forces across the cellulose membrane. In general, this promoted a more intense mineral precipitation on the phosphate membrane surface. A strong deposition of calcium phosphate mineral was observed at pH 9.0 as a primary layer with a homogeneous microstructure, being totally absent at pH 6.0. The membrane showed an additional crystal phase at pH 7.0 exhibiting a very particular hexagonal or cuttlebone shape, mainly on the phosphate surface. When organic fluids of A. cygnea were included, these crystal forms presented a high tendency to aggregate under rosaceous shapes, also predominantly in the phosphate side. The cellulose membrane was permeable to small organic molecules that diffused from the calcium towards the phosphate side. In the calcium side, very few similar crystals were observed. The presence of organic matrix from A. cygnea fluids induced a preliminary apatite-brushite crystal polymorphism. So, the present results suggest that cellulose membranes can be used as surrogates of biological epithelia with preferential ionic diffusion from the calcium to the phosphate side where the main mineral precipitation events occurred. Additionally, the organic fluids from freshwater bivalves should be also thoroughly researched in the applied biomedical field, as mineral nucleators and crystal modulators on biosynthetic systems.

Microstructural Characterization of Inner Shell Layers in the Freshwater BivalveAnodonta cygnea

The shell microstructure of freshwater bivalve Anodonta cygnea was observed from the ventral margin toward the intrapallial regions by scanning electronic microscopy during the spring/summer period. The three main structures—the periostracum, prismatic, and nacreous layers—were visualized in a sequential path from the ventral margin toward the intrapallial regions. Although with always the same calcium carbonate polymorph (aragonite), A. cygnea presented composites of aragonite crystals embedded within an organic matrix displaying varied morphologies and structural arrangements. The prisms of the prismatic shell layer are composed of single-crystal fibers radiating from spheruliths, whereas in the nacreous layer the structure is similar to that of a brick wall, with tablets in each layer offset with respect to those in the layers above and below them. From the shell ventral margin, the last nacreous lamina formed exhibit crystals with rounded forms, followed by rhombic and hexagonal shapes toward the interior with irregular microstructure. There are gradual intermediate forms between these distinct shapes. This study offers a description of the inner calcareous mineralized layers of A. cygnea and will be useful for future comparative studies in several research areas such as phylogenetic evolution, ecotoxicology, and the influence of environmental and physicochemical factors on the shell formation process in freshwater bivalves.

Effects of Cd2+ on the calcium metabolism and shell mineralization of bivalve Anodonta cygnea

The survival of freshwater mussels is affected by cadmium ions because they accumulate in tissues and fluids and displace calcium ions in transport and regulation mechanisms. A preliminary study of the freshwater bivalve Anodonta cygnea under enriched Ca2+ (2 mM) water for different periods (3, 4, and 18 days), showed significant increase in calcium levels (p < 0.05) in the ventricular fluid with concentration proportional to time of exposure. Groups of A. cygnea were then exposed to different treatments for 4 days: Group 1 to Ca2+ (2 mM) enriched water as a control; Group 2 to a sub-lethal concentration (10−6 M) of Cd2+; Group 3 to an injection of vitamin D (1340 IU mL−1); Group 4 to an injection of SEA0400 (950 µg mL−1), a sodium/calcium exchanger (NCX) inhibitor. Fluid samples from different compartments of the animals were analyzed for Ca2+ and Cd2+ composition as well as for other ions relevant to the osmoregulation (Na+, Mg2+, K+, and Cl−) and their osmolarity. The treatment groups had significantly higher calcium concentrations in their ventricular fluid (p < 0.05) under Cd2+ and vitamin D, also in pericardium fluid under Cd2+, vitamin D, and SEA0400. We infer a specific transport mechanism from the intestinal to the ventricular fluid that may be induced by vitamin D. A general internal acidosis by Cd2+, releasing calcium from the calcareous concretions and a specific inhibition of calcium resorption, by the SEA0400, from the pericardium towards the ventricle is discussed. Cd2+ incubation significantly augmented (p < 0.05) the cadmium contents of all experimental fluids and tissues. Scanning electron microscope (SEM) imaging of the border regions of the shell showed clear ultrastructural changes in respone to Cd2+ incubation, suggesting a direct influence of this ion on the shell biomineralization mechanisms.

Studies on a PMCA-like protein in the outer mantle epithelium of Anodonta cygnea: insights on calcium transcellular dynamics

Early studies on the outer mantle epithelium (OME) cells of the freshwater bivalve Anodonta cygnea (Linnaeus, 1758) revealed high ionic calcium concentrations by electrophysiological methods and subsequently a high tendency to reach an intracellular toxic condition. This toxicity could be neutralized by specific mechanisms in the cytosol of OME cells of A. cygnea. The present immunocytochemistry studies of OME cells by light and transmission electron microscopy (TEM) clearly showed a positive reaction of an antibody directed against the human plasma membrane Ca(2+)-ATPase 1 (PMCA-1) in the cytoplasm of OME cells. Also, western blot analysis of different fractions of OME cells with anti human PMCA-1 and C28R2 antibodies confirmed the presence of a PMCA-like protein with an unusual topographical localization and a molecular weight of only 70-80 kDa. These results lead us to speculate that this PMCA-like protein is distributed either in the plasma membrane or in the entire cytosol, where it eventually regulates intracellular calcium levels. Interestingly, the antibody reactions showed seasonal variations, being highest in OME samples prepared during summer when A. cygnea live under natural acidosis and absent in samples taken in winter conditions, which is in accordance with the seasonal variation of shell calcification rates. During winter, PMCA-1 antibody reaction was also detected in OME cells of animals kept only under experimentally induced acidosis conditions. Therefore, we assume that a functional role for this PMCA-like protein in the intracellular calcium regulation of OME cells during the mineralization of the shells of A. cygnea can be speculated.

Do Zebra Mussels Grow Faster on Live Unionids than on Inanimate Substrate? A Study with Field Enclosures

The zebra mussel Dreissena polymorpha has invaded numerous freshwaters in Europe and North America and can foul many types of solid substrates, including unionid bivalves. In field experiments we compared growth rates of dreissenids on live specimens of the freshwater bivalve Anodonta cygnea to growth rates of dreissenids on stones. Dreissena density in the study lake was about 1000 m–2 in most places, Anodonta density approximately 1 m–2 and about 50% of the Anodonta were infested with 10–30 Dreissena . In summer/autumn small dreissenids generally grew faster on live Anodonta than on stones. Similar trends were observed for spring, but differences of growth increments between dreissenids on live Anodonta and stones were usually not significant. Dreissenids settled down or moved towards the ingestion/egestion siphons of Anodonta and ingestion siphons of dreissenids were directed towards siphons of Anodonta . These results suggest that dreissenids can use the food provided by the filter current of the large Anodonta . (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Biomonitoring of the Vltava River radioactivity

In 1996-2000, before the nuclear power plant starting in South Bohemia (Czech Rep.), the monitoring of the Vltava River, where the liquid radioactive effluents will be discharged, was accomplished. The biomonitoring of the hydrosphere was made by means of water plants and freshwater bivalve. Under given conditions, the water mosses Hygrohypnum ochraceum and Fontinalis antipyretica seem to be the most suitable plants. As to riverbank plants, Polygonum hydropiper species and the reed Glyceria maxima and Phalaris arundinacea were chosen. The freshwater bivalve Anodonta cygnea (L). Was also sampled. The 137 Cs mass activity values of the hydrobionts range from 100 to 101 Bq.kg-1 dry weight and its time course for the plants is descending: the ecological half-life estimations range from 1 to 5 years. As to natural radionuclides: 40 K prove the highest values of mass activity (of the order of 102 Bq.kg-1 d. w.), while the radionuclides of actinium series prove the lowest ones (often under the detection limit). The radionuclides of uranium and thorium series are between these two extremities. The radioactivity of bivalve is low and practically constant during the five years period. The biomonitoring of the hydrosphere will be carried on after NPP running in 2001.

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 μg l −1 (9.25 and 18.5 μM 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 μg l −1 added Al produced no detectable change, while 500 μg 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 μg l −1 added Al than in those exposed to 500 μg 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 μg 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.

Effects of episodic low pH exposure on the valve movements of the freshwater bivalve Anodonta cygnea L.

Adductor activity of the freshwater mussel Andonta cygnea was examined continuously for a period of 32 days under circumneutral and acidic conditions. The adductor activity and valve movements were monitored using a Mussel Observation Program (MOP) based on electromagnetic induction. Acute acid exposure disturbed the rhythmic valve activity by changing the normal pattern of successive active/inactive (open/closed) periods. Fluctuations in water pH were reflected in changes in mussel adductor activity. Obviously these changes in adductor activity induced by low pH have adaptive significance for mussels in water subject to episodic acidification.

Glycogen degradation and end products of anaerobic metabolism in the fresh water bivalveAnodonta cygnea

1. Glycogen levels and the production of metabolic end products by the fresh water bivalveAnodonta cygena were determined under aerobic and anaerobic conditions. 2. The glycogen content accounts for 5–14% of the dry weight. After 30 hours of aerobic or anaerobic incubation no significant glycogen breakdown could be observed. 3. L-lactate and alanine seem to be only initial end products, whereas succinate increases continuously during the first 15 hours. 4. Anodonta cygnea also accumulates volatile fatty acids during anaerobiosis. After 30 hours of anoxia 30.1 and 46.0 μmol g−1 dry weight of acetate and propionate respectively are formed. In fact the volatile fatty acids account together for about 75% of the total accumulated end products.

Partial purification and properties of octopine dehydrogenase and the formation of octopine inAnodonta cygnea L.

1. Octopine dehydrogenase was purified 120-fold from adductor muscle of the fresh water bivalveAnodonta cygnea L. by gel filtration on Sephadex G-100 and chromatography on DEAE-Sephadex A 50 and hydroxylapatite. 2. In comparison to the purified enzyme from marine species the final preparation of the octopine dehydrogenase fromAnodonta showed similarK m (1.0 mM for arginine, 0.4 mM for pyruvate, 1.2 mM for octopine) and pH values (6.3 and 10.1 respectively) and a similar molecular weight (40 000 Dalton). 3. During electrophoresis on standard polyacrylamide gels octopine dehydrogenase showed a multiple band pattern which is due to isoenzymes. 4. Alanine, lactate and succinate had no effect on the enzyme activity. Octopine, however, inhibited it strongly (Fig. 4) 5. Thede-novo synthesis of octopine in isolated adductor muscles from14C-pyruvate was not significantly different under aerobic or anaerobic conditions.

Observations on the Postanaerobic Metabolism of Some Fresh-Water Snails

Previous articleNext article No AccessObservations on the Postanaerobic Metabolism of Some Fresh-Water SnailsTheodor Von Brand, Patricia McMahon, and M. O. NolanTheodor Von Brand Search for more articles by this author , Patricia McMahon Search for more articles by this author , and M. O. Nolan Search for more articles by this author PDFPDF PLUS Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by Volume 28, Number 1Jan., 1955 Article DOIhttps://doi.org/10.1086/physzool.28.1.30152177 Views: 8Total views on this site Citations: 14Citations are reported from Crossref Journal History This article was published in Physiological Zoology (1928-1998), which is continued by Physiological and Biochemical Zoology (1999-present). Copyright 1955 The University of ChicagoPDF download Crossref reports the following articles citing this article:J.D. Thomas, C. Kowalczyk, B. Somasundaram The biochemical ecology of Biomphalaria glabrata, a snail host of Scistosoma mansoni: Short chain carboxylic and amino acids as phagostimulants, Comparative Biochemistry and Physiology Part A: Physiology 93, no.44 (Jan 1989): 899–911.https://doi.org/10.1016/0300-9629(89)90516-1Theodorus C.M. Wijsman, Hans C. van der Lugt, Hans P. Hoogland Anaerobic metabolism in the freshwater snail Lymnaea stagnalis: Haemolymph as a reservoir of d-lactate and succinate, Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 81, no.44 (Jan 1985): 889–895.https://doi.org/10.1016/0305-0491(85)90084-7 Uptake and assimilation of short chain carboxylic acids by Biomphalaria glabrata (Say), the freshwater pulmonate snail host of Schistosoma mansoni (Sambon), Proceedings of the Royal Society of London. Series B. Biological Sciences 222, no.12291229 (Jan 1997): 447–476.https://doi.org/10.1098/rspb.1984.0077R.L. Patience, J.D. Thomas, P.R. Sterry Production and release of carboxylic acids during oxic and anoxic metabolism by the pulmonate snail Biomphalaria glabrata (Say), Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 76, no.22 (Jan 1983): 253–262.https://doi.org/10.1016/0305-0491(83)90067-6Ming-Shan Ho, Paul L. Zubkoff Volatile fatty acids of the ribbed mussel, Geukensia demissa, Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 74, no.33 (Jan 1983): 539–542.https://doi.org/10.1016/0305-0491(83)90225-0J.D. Thomas, J. Ofosu-Barko, R.L. Patience Behavioural responses to carboxylic and amino acids by Biomphalaria glabrata (say), the snail host of Schistosoma mansoni (Sambon), and other freshwater molluscs, Comparative Biochemistry and Physiology Part C: Comparative Pharmacology 75, no.11 (Jan 1983): 57–76.https://doi.org/10.1016/0742-8413(83)90012-9Arend A. Dogterom The effect of the growth hormone of the freshwater snail Lymnaea stagnalis on biochemical composition and nitrogenous wastes, Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 65, no.11 (Jan 1980): 163–167.https://doi.org/10.1016/0305-0491(80)90126-1Gerd Gäde, Hans Wilps, Jacques H. F. M. Kluytmans, Albertus de Zwaan Glycogen degradation and end products of anaerobic metabolism in the fresh water bivalveAnodonta cygnea, Journal of comparative physiology 104, no.11 (Dec 1975): 79–85.https://doi.org/10.1007/BF02482839P.E. Haeser, F.B. De Jorge Anoxic anoxia in strophocheilus (pulmonata mollusca), Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 38, no.44 (Apr 1971): 753–757.https://doi.org/10.1016/0305-0491(71)90276-8Robert H. Catlett Rapid acclimation and oxygen consumption of the rough limpet,, Life Sciences 10, no.66 (Mar 1971): 355–359.https://doi.org/10.1016/0024-3205(71)90077-4 References, (Jan 1967): 345–390.https://doi.org/10.1016/S0065-2881(08)60334-XW. RUSSELL HUNTER Physiological Aspects of Ecology in Nonmarine Molluscs, (Jan 1964): 83–126.https://doi.org/10.1016/B978-1-4832-3241-6.50009-3Eugene C. Weinbach, M.O. Nolan The effect of pentachlorophenol on the metabolism, of the snail Australorbis glabratus, Experimental Parasitology 5, no.33 (May 1956): 276–284.https://doi.org/10.1016/0014-4894(56)90040-6Walter L. Newton, Theodor von Brand Comparative physiological studies on two geographical strains of Australorbis glabratus, Experimental Parasitology 4, no.33 (May 1955): 244–255.https://doi.org/10.1016/0014-4894(55)90028-X