Low sensitivity of ciliary activity in the gills of Anodonta cygnea to some ecotoxicals

1. The effects of some ecotoxical model substances on the activity of frontal gill cilia of freshwater mussel Anodonta cygnea were studied in 1 and 24 hr experiments with the particle transport method. The effects were compared with the results of standard Daphnia magna ec₅₀ tests with the same substances.2. Variations of pH between 4.8 and 8.1 affected the ciliary activity by less than 30%.3. Ciliary activity was less sensitive to heavy metal salts, sodium lauryl sulphate and to two pulp industry waste model substances than D. magna.4. Compared to D. magna, the ciliary activity was more sensitive to formalin and 2,4-dinitrophenol, and as sensitive to ethanol.5. Ciliary activity in bivalve gills is a well regulated and/or a relative insensitive function, but it may be used as a rapid screening test for some cell ecotoxicals.     

In vitro culture of glochidia from the freshwater mussel Anodonta cygnea

Abstract. Larvae of the freshwater swan mussel, Anodonta cygnea , were cultured in artificial media at the controlled temperature of 23°±2°C, with successful metamorphosis for the first time. The artificial medium contained a mixture of M199, common carp plasma, and antibiotics/antimycotics. Glochidia were reared to the juvenile stage in the medium after 10–11 d of culture. After 15 d of controlled feeding with phytoplankton, the juveniles showed an elongated shell with several growth lines. Larval survival was 34.3±9.3%, whereas the proportion undergoing metamorphosis was ≤60.8±4.2%. The ultrastructure of early developmental stages was observed by scanning electron microscopy, from the glochidial to the juvenile stage. Glochidia had a hooked shell, with two equal triangular valves formed by a calcareous layer with numerous pores and covered by a thin cuticle of chitin–keratin. The appearance of the complete foot within 11 d of in vitro culture was considered the final feature of metamorphosis to the juvenile stage. The main alteration during juvenile development was the formation, under the glochidial shell, of a new periostracum with growth lines. The prominent foot, gradually covered by long, dense cilia, showed rhythmical movements involved in the capture of particulate matter. Similarly, cilia and microvilli present in the mantle also performed the same role. Longer cilia, sparsely distributed in the mantle, may function as chemotactile sensors.     

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.     

Effect of sub-lethal concentrations of aluminium on the filtration activity of the freshwater mussel Anodonta cygnea L. at neutral pH

Significant amounts of aluminium (Al) are commonly present in rivers and lakes, largely in particulate form in neutral waters. Freshwater bivalves, as filter feeders are therefore exposed to both particulate and dissolved metal and are potentially vulnerable to Al. The effect of Al on filtering behaviour of the freshwater mussel Anodonta cygnea L. was investigated during short (1 hour) and long-term (15 days) exposure to environmentally relevant concentrations (250 and 500 microg l(-1)) at neutral pH. Water flow through the outflow siphon was monitored as an indicator of pumping capacity. Short-term (1 hour) exposure to 500 microg l(-1) added Al produced an irreversible decrease in the duration of filtering periods, presumably as an avoidance response to the toxicant. One-hour exposure 250 microg l(-1) Al had no detectable effect. When mussels were exposed to 250 or 500 microg l(-1) added Al for 15 days, siphon activity measured in days 11-15 of exposure was inhibited by 50% and 65%, respectively, compared to pre-exposure levels. Recovery occurred following transfer of mussels to uncontaminated water. Interaction between Al and freshwater bivalves at neutral pH may affect both the performance of the mussels and the chemical speciation of the metal in the natural environment.     

Effect of some insecticides on the periodic activity of the fresh-water mussel (Anodonta cygnea L.).

The effect of the insecticides Hungaria L-7, Dimecron-50, Thimet-10 G and Satox 20 WSC containing lindane, phosphamidon, phorate and trichlorfon, respectively, as active ingredient was investigated on the survival and pumping behaviour of the fresh-water mussel Anodonta cygnea L. The shortening of active periods was used as indicator of the sublethal effects. Hungaria L-7, Thimet-10 G and Satox 20 WSC caused significant inhibition of the filtering activity, while Dimecron-50 was less effective. Fifty per cent shortening of active periods required 6 X 10-3 g/l Hungaria L-7, 8 X 10-3 g/l Thimet-10 G and 2 X 10-2 ml/l Satox 20 WSC. Thimet and Satox were lethal at 10-1 g/l and 1 m/l concentrations, respectively. Although the mechanism of the effect can be different with various substances, the results have shown, that monitoring of the activity of mussels can be a suitable test for the biological indications of water pollution.     

The role of abiotic factors and pesticide levels on enzymatic activity in the freshwater mussel Anodonta cygnea at three different exposure sites

Natural variation in abiotic factors, such as temperature and pH, probably influence the activity of enzymes used as potential biomarkers in bivalve mollusks to assess environmental contamination in the field. Changes in levels of an enzymatic biomarker may thus merely reflect natural variation in the annual physiological cycle of a species rather than exposure to contaminants. To investigate this issue, we documented the relationship between pesticide levels in water and three different enzymatic biomarkers over 1 year in enclosed populations of the freshwater unionid mussel Anodonta cygnea at three different sites of exposure. We considered the natural variation in temperature, pH and dissolved oxygen over the year and across the different sites as a potential correlate of enzymatic activity to disentangle the relative contribution of abiotic factors and pesticide levels. Pesticide levels varied among the three sites and over the course of the year. Catalase (CAT) and acetylcholinesterase activity (AChE) varied as a function of abiotic factors but showed no relation to pesticide levels. Glutathione S-transferase (GST) activity was also related to abiotic factors but also decreased with increases in total pesticide levels. The lack of activity induction or inhibition by pesticides and the natural variation in abiotic factors among sites and across time limits the use of CAT and AChE to assess environmental contamination in this species.     

Factors affecting the heart activity and blood pressure of the swan mussel Anodonta cygnea.

1. The rate of heart beat increased with temperature and was three times as high in the active as in the inactive animal. 2. The rate of shell valve movement rose and the rate of heart beat fell when the foot was extended. 3. The rates of heart beat and shell valve movement decreased when the water was saturated with carbon dioxide. This heart response remained when the visceral ganglion was destroyed. 4. Ventricular contraction occurred simultaneously over the shole chamber. The passage of blood into the posterior aorta could be restricted by the protuberances on its wall. 5. Pericardial cavity pressure rose by about 5 cm H2O at shell valve adduction and 0-25--0-6 cm H2O at ventricular diastole. 6. Pulse pressure changes of 0-25--0-6 cm H2O occurred in the auricle and 1--3 cm H2O in the ventricle and anterior aorta.     

Light and Electron microscopic autoradiography of 3H-dopamine uptake in the ganglia of the freshwater mussel, Anodonta cygnea L

In the central nervous system of the freshwater mussel, Anodonta cygnea, uptake of 3H-dopamine has been investigated by light and electron microscopic autoradiography following in vitro incubation and topical application, respectively. The autoradiographic reaction observed over the ganglia can be taken as a morphological evidence of the existence of a specific uptake mechanism for dopamine. It has been found that labeled dopamine was taken up by the axon profiles and terminals, while perikarya remained free of activity, indicating a basic difference between the perkaryal and axonal membrane in respect of the 3H-dopamine uptake. According to the electron microscopic autoradiography, labeled terminals and axon profiles contained both normal and accentric dense-core vesicles and they proved to be similar to those seen after 3H-serotonin in vitro incubation. Therefore, it seems possible that certain Anodonta central neurons contain both dopamine and serotonin.     

Changes in gape frequency, siphon activity and thermal response in the freshwater bivalves Anodonta cygnea and Margaritifera falcata

Physiologically-driven rhythms in bivalve molluscs are predictedto vary as a function of metabolic rate and temperature, incontrast to genetically predisposed biological clocks. Theserhythms can be evaluated using long-term video monitoring techniquesunder controlled conditions in laboratory aquaria. The bivalvesAnodonta cygnea and Margaritifera falcata were used to evaluatethe effect of temperature on rhythms in gape and the formationof siphons at the mantle edge. Frequency and duration of shellclosure vary with temperature in both species, but with differentresponses. Mean duration of intervals of valve closure decreasesas temperature rises in both species, and is consistent withphysiological limitation by increased biological oxygen demand.For A. cygnea, cumulative gape duration peaks at 25°C, withless time spent closed than at any other temperature, but increasingtemperatures correspond to an increase in gape frequency witha strong increase observed at 31°C. In contrast, frequencyof adduction and valve closure peak at 25°C in M. falcata,and continuous gaping is observed above 29.5°C. This physiologicalstress is consistent with evidence from sclerochronologically-calibratedstable isotope studies of shells, where growth breaks in manymarine taxa coincide with maximum temperatures above 31°Cas derived for ¹⁸O_carbonate. The results of this study suggestthat these growth breaks may be due to physiological limitationsin oxygen uptake and metabolic activity, rather than being adirect consequence of elevated temperature alone. (Received 17 March 2008; accepted 3 October 2008)     

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.     

Morphological and chemical characterization of mineral concretions in the freshwater bivalve Anodonta cygnea (Unionidae)

The freshwater mussel Anodonta cygnea is commonly used as a model organism for biomineralization studies, its peculiar morphofunctional properties also make it an excellent environmental biomonitor. The first detailed on the calcareous concretions from gill and mantle tissue, as well as fluids of the freshwater bivalve A. cygnea, supported by histological, scanning, spectrometry, and spectroscopy analyses. Through these analyses, the morphology, structure, and chemical characterization of these biomineral concretions were accomplished. The concretions represent a high percentage of the dry weight of these organisms. In gill tissue, it can reach up to 50% of dry weight prior to reproductive maturity. Analysis of elemental composition of the tissue concretions showed the presence of calcium and phosphate, as main components, associated with other residual elements like iron, manganese, magnesium, and zinc. Concretions are arranged in concentric alternated layers of organic and inorganic matrix. The shape and size of the concretions vary substantially, from very small, less than 1 μm diameter with very regular round structure, found mainly in the mantle tissue, to more than 50 μm length with irregular globular clusters, found predominantly in the gills. The microstructural organization is of a hydroxyapatite polymorphism in the mantle, in contrast to the gills, which exhibit irregular structure and carbonated hydroxyapatite polymorphism. These differences are supported by higher contents of dinitrogen pentoxide, magnesium, and iron in the mantle concretions, but higher contents of manganese and zinc in the gills. Furthermore, the results indicate that the mineral concretion formation in A. cygnea is a hemocytes reaction to particle or toxic invasions. A second relevant role, concerns the close involvement of these microspherules on the adult and larval shell calcification. J. Morphol. 276:65–76, 2015. © 2014 Wiley Periodicals, Inc.     

Thermal resistance of the ciliary activity in the gills of the fresh water mussel Anodonta anatina

1. 1.|The thermal resistance of the activity of frontal cilia in the median gills of the fresh water mussel Anodonta anatina was studied.

2. 2.|The resistance acclimation appeared in 2 days in the gills of intact animals, but not in the isolated gills kept at 4, 14 and 24°C, for between 1 to 3 days.

3. 3.|Warm-acclimation increased the ACh sensitivity of the gills of intact mussels.

4. 4.|Isolation of the gills enhanced the thermal resistance.

5. 5.|ACh, choline and tetramethylammonium enhanced the thermal resistance in the isolated gills. whereas atropine and physostigmine diminished it.

6. 6.|It is concluded that in A. anatina the control if the thermal resistance is probably neural.

Beta-adrenoreceptors in the muscular tissue of Anodonta cygnea

The beta-adrenoceptors in the muscular tissue of Anodonta cygnea have been studied for the first time with the use of antagonist [125I] iodocyanopindolol. The tissue membrane had only one class of binding sites with Kd 2.9 +/- 0.02 pM and the maximal number of binding sites (Bmax) 110 +/- 2.4 fmoles/mg of protein. The potency of beta-agonists and antagonists for displacing [125I] iodocyanopindolol for its beta-AR complexes was the following: isoproterenol greater than adrenaline greater than noradrenaline-propranolol greater than serotonin much greater than dopamine greater than phentolamine. The GTP negative regulation of beta-AR affinity has been found. The data obtained show that the beta-AR are functionally coupled with GTP-binding protein which were similar to GTP-proteins of vertebrates.     

An electron microscope study of the formation of the periostracum in the freshwater bivalve, Anodonta cygnea

The periostracum and cells lining the periostracal groove of Anodonta cygnea L. have been studied at the electron microscope level. The cells lining the inner face of the outer fold differ in fine structural details, five cell types being recognized. Along the length of the outer surface of the middle fold, to which the periostracum is closely applied, only two cell types are evident. At the base of the periostracal groove the two epithelia are separated by a bulbous region containing a group of basal cells which initiate the periostracum. The periostracum, which is homogenously electron-lucid, originates in the intercellular space between a basal cell and the first cell of the middle fold. It increases in thickness in the periostracal groove due to the secretory activity of the different outer fold cells. The cells of the middle fold do not appear to be involved in periostracum formation.