Arcid Clam Research Articles

The alien bivalve Anadara demiri (Arcidae): a new invader of the Adriatic Sea, Italy

This paper reports the observations recorded on the arcid clam, Anadara demiri, from its first reporting in the Adriatic Sea three years ago, when its presence only was noted, to the present day when its invasive potential is confirmed. Repeated recruitment events were observed and confirmed by the co-existence of two or more cohorts within the population. Anadara demiri appears to be a year-round spawner with two spawning peaks (late winter and summer). Together with the congeneric invader, Anadara inaequivalvis, it has colonized the external belt of the fine well-sorted sand biocoenosis where they may numerically dominate the benthic community. Anadara demiri appears to have gained full status as a component of the Adriatic Sea coastal benthic community despite its very recent first entry. Numerous features make it a strong competitor among which is the ability, retained into adult life, of attaching, by means of byssus threads to all kinds of hard substrata, particularly to other live molluscs such as the gastropod Aporrhais pespelecani and the bivalves Anadara inaequivalvis and Chamelea gallina. The presence of species actively interfering with A. demiri in various ways (the polychaetes, Polydora sp. and Sabellaria spinulosa, that bore into and foul the shell, respectively, and the predatory asteroid Astropecten irregularis) is discussed.

Primary Structure of Noetia ponderosa Hemoglobins: Functional Correlates

ABSTRACTHomo- and heterodimeric hemoglobins have been isolated from the red cells of the arcid clam Noetia ponderosa (Np). These hemoglobins bind oxygen cooperatively. An extensively studied dimeric hemoglobin from another arcid clam, Scapaharca inaequivalvis, exhibits a molecular mechanism for cooperative ligand binding that is radically different from tetrameric vertebrate hemoglobins. In this study, the two chains found in both Noetia hemoglobins are sequenced and compared to the hemoglobins of the related clam S. inaequivalvis to determine whether Noetia hemoglobins have the structural basis for the same unusual mechanism for cooperative ligand binding and to inquire about the structural basis of absence of tetramers. Although the Noetia sequences are homologous to the Scapharca sequences, critical differences exist. The lack of tetramerization of Np subunits is most likely related to the absence of critical residues in the A and G helices that stabilize the interdimer contact seen in the Scapharca Hb tetramer. The lower affinity of the homodimer (Np-I), but particularly the heterodimer (Np-II) with respect to the homodimer and heterotetramer of Scapharca, can be due to (i) changes in the proximal heme environment and (ii) changes in the dimer interface. Interactions between Asn 100 and the heme of the other subunit are altered in Np-II due to the substitution of this residue by methionine, possibly causing the reduced O2 affinity of the heterodimer of Noetia. (iii) Sequence changes in the E and F helices present in Np-I and Np-II could also contribute to the effect through interfacial changes. In particular, the substitution of Val for Thr in position 72 is expected to have a substantial influence on the interface. We conclude that Np dimers have the structural basis for a direct heme–heme interaction mechanism for cooperativity, as in Scapharca, but there are enough sequence changes to suggest that the pathway of interaction might be somewhat different.

Cadmium accumulation and Cd-binding proteins in the bivalve Scapharca inaequivalvis

In the arcid clam, Scapharca inaequivalvis, cadmium was accumulated in the kidneys, gills, viscera, mantle, foot, muscle and red blood cells. Maximum levels were reached in the kidney (725.70 μ/g dry weight) after 4 weeks exposure to 0.5 μg Cd/ml sea-water. The accumulation was higher during the warm rather than in the cold season and in animals exposed to 0.5 as compared with 0.05 μg Cd/ml sea-water. Concentrations of zinc and copper were unaffected by cadmium treatment, whereas iron levels changed during cadmium exposure. When molluscs were returned to natural sea-water, cadmium levels fell slowly. The metal was found in the cytosolic fraction, bound to a low molecular weight ligand, shown by gel filtration to be a Cd-protein with an apparent molecular weight of 10,000 Da, possibly a metallothionein. Ion exchange chromatography of this protein isolated from the viscera revealed the presence of at least two Cd-binding isoforms; the -SH: Cd ratio of the main isoform was 2.8, the UV spectra presented a strong absorption band at 250 nm.

Heme-heme interactions in a homodimeric cooperative hemoglobin. Evidence from transient Raman scattering.

The comparison of the resonance Raman spectrum of the deoxy dimeric hemoglobin (HbI) from the Arcid clam, Scapharca inaequivalvis, to its CO photoproduct at 10 ns reveals several significant differences in the low frequency vibrational modes including those involving motions of the peripheral substituents on the heme such as the propionates. This finding reflects the involvement of the propionates in a hydrogen bonding network which connects the two heme groups across the subunit interface and is sensitive to ligand binding. A frequency difference in the iron-histidine stretching mode of 6 cm-1 in this invertebrate hemoglobin is substantially smaller than that detected in tetrameric vertebrate hemoglobins under similar conditions. Time evolution studies of the vibrational modes from the photoproduct demonstrate that the transient form relaxes to the deoxy structure concertedly with a half-life of 1 microseconds, the time scale in which tertiary relaxations of tetrameric hemoglobins occur. These data establish that the tertiary and quaternary structural changes take place on the same time scale and confirm a mechanism of cooperativity involving direct interaction between the two heme groups through the peripheral substituents. This direct structural communication provides a very tight linkage between the heme groups prohibiting modulation of the oxygen affinity by factors in the physiological milieu.

The 2.4-Å crystal structure of Scapharca dimeric hemoglobin: Cooperativity based on directly communicating hemes at a novel subunit interface

The crystal structure of the cooperative dimeric hemoglobin from the arcid clam, Scapharca inaequivalvis, has been determined in the carbonmonoxy state. The phase problem was solved for reflections with Bragg spacings greater than 3 A using anomalous scattering from the porphyrin iron atoms measured at a single wavelength in combination with molecular averaging. The model built into this electron density map has been refined at 2.4 A resolution by means of stereochemically restrained least squares minimization to a conventional R-value of 0.156. The root mean square deviation from ideal bond lengths and angles are 0.013 A and 1.7 °, respectively. In addition to the 2336 hemoglobin atoms, 214 water molecules have been incorporated into the model. This structure reveals the details of an assemblage of two identical myoglobin-like subunits that is radically different from vertebrate hemoglobins. The subunit interface is formed by direct apposition of the E and F helices, whereas these surfaces are external in vertebrate hemoglobins. The interface has both hydrophobic and hydrophilic character. Two symmetrically related hydrophobic regions are formed between subunits. Six residues are involved in each of these regions that pack tightly enough to exclude water but have only a few atoms in close van der Waals contact. A number of ordered water molecules line the interface and form bridging hydrogen bonds between subunits. Four intersubunit ionic interactions are formed, two of which involve negatively charged propionate groups of the porphyrin. In contrast to cooperative vertebrate hemoglobins, a hydrogen bond network provides a direct route for communication between the two heme groups.

Anadara ovalis hemoglobins: Distinct dissociation and ligand binding characteristics

1. 1. The red cells of the arcid clam Anadara ovalis contain two electrophoretically distinct hemoglobins: Hb Major (Hb Ma) and Hb Minor (Hb Mi). The major component consists of two electrophoretically indistinguishable tetramers each composed of two heterodimers; the minor hemoblogin is a homodimer whose subunits are different from the tetramer. 2. 2. Functionally, Hb Ma has a higher P 50, exhibits a concentration dependent oxygen affinity, has significant ligand cooperativity ( n = 2.0), lacks a Bohr effect and is unaffected by ATP. 3. 3. HB Mi has a P 50 which is lower and independent of hemoglobin concentration, shows appreciable cooperativity ( n = 1.4) and exhibits no heterotropic effects. 4. 4. Both Hb Ma and Mi are resistant to dissociation in the presence of 1.0M NaI, NaCl and guanidine-HCl but dissociate to monomers when converted to the aquamet but not the cyanmet derivative. 5. 5. The dissociation is completely inhibited by mercaptoethanol. 6. 6. The large number of reactive - SH groups (10–13 per tetramer) suggests that the monomerization is mediated by intra-subunit disulfide bridge formation.

Effect of environmental parameters on lysosomal marker enzymes in the tropical blood clam Anadara granosa

The lysosomal marker enzymes, arylsulfatase and acid phosphatase, in a tropical burrowing arcid clam Anadara granosa L. have been found to exhibit seasonal variations. The activity of both enzymes decreased with increase in ambient temperature and fell with increase in salinity. Lysosomal latency for these enzymes, however, was not significantly affected by environmental parameters, including salinity, temperature, nutritional status, breeding season, etc. The physico-chemical characteristics of the habitat were found to induce reversible changes in the lysosomal latency and enzyme activity. Exposure to elevated temperature (ΔT=7° to 8°C), towards the upper limit of its physiological tolerance, was found to labilize the lysosomes. The biochemical methods optimized for the demonstration of lysosomal stability under exposure to environmental stressors could also be extended to quantify the impact of various pollutants, including heavy metals, radionuclides, etc. at the subcellular level.

The quaternary structure of an unusual high-molecular-weight intracellular haemoglobin from the bivalve mollusc Barbatia reeveana.

The arcid clam Barbatia reeveana contains an intracellular haemoglobin with an unusual structure. First, compared with other intracellular haemoglobins, it is extremely large, with a mol.wt. of 430000 and an s(20,w) of 13.6S. A minor component (mol.wt.=220000; s(20,w)=9.7S) is also present as a probable dissociation product of the major component. Secondly, this haemoglobin has an unusual subunit structure. It contains 1mol of haem per 16000g of protein, in common with most other haemoglobins. However, the smallest polypeptide that could be obtained after treatment with sodium dodecyl sulphate or 6m-guanidine with reducing agent has a mol.wt. of 32000-37000. Digestion of the haemoglobin with the proteinase subtilisin produces both 57000- and 30000-mol.wt. aggregates that contain 1mol of haem per 16000g of protein and that can be dissociated into 16500-mol.wt. polypeptides by treatment with sodium dodecyl sulphate. The intact polymer shows slight co-operativity (h=1.7), lacks a Bohr effect between pH7 and 8, and has a low oxygen affinity [P(50)=4.8kPa (36mmHg) at 20 degrees C] relative to other haemoglobins. The 30000-mol.wt. aggregate obtained by digestion of the polymer binds oxygen reversibly with an affinity greater than that of the polymer, but with some co-operativity (h=1.7). These results are consistent with the hypothesis that the subunits of this unusually large intracellular haemoglobin are 32000-mol.wt. polypeptides that in turn are composed of two covalently linked haem-containing oxygen-binding domains. This is the first report of an intracellular haemoglobin with such a structure.

Flux of certain radionuclides in the blood-clam Anadara granosa Linneaus under environmental conditions

The flux of fission product nuclides, caesium-137. cerium-144. and ruthenium-106, in the Bombay harbour ecosystem has been measured over a period of six years (1970–1976). during which the input of controlled low level radioactive waste was gradually reduced by more than 20 times. Of the various pelagic and benthic species studied, the arcid clam Anadara granosa Linn, showed a specific affinity for all three radionuclides; sequence of the degree of bio-accumulation was Ce ⩾ Ru > Cs. Furthermore, the concentration of these nuclides in the tissues varied with the levels in the sedimentary material, which in turn was related to the inputs, maintaining some kind of equilibrium. On reaching maximum accumulation the levels of all three nuclides in the clam population decreased exponentially with time although the radioactivity was still available especially in the sediment. This decrease of radioactivity is defined as an ‘ecological loss rate’. The ecological half-time for cerium-144 and ruthenium-106 was about 3.6 times longer, and that of caesium-137 significantly shorter, than their respective physical half-lives. The ecological half-times of all three radionuclides were predominantly influenced by the environment rather than by the biological half-lives. This indicates that the biological half-lives of these nuclides were much shorter than their respective ecological half-lives. The biological half-time of caesium-137 was biphasic; the short-lived component had a half-time of about three days and the long-lived component had one of about 15 days. The bio-accumulation of an activation product nuclide cobalt-60 in A. granosa increased with the time of exposure and reached a level significantly higher than that of caesium-137, but was independent of the ambient concentrations and temperature. Thus the species was found to maintain the tissue concentration of fission product nuclides, cerium-144, ruthenium-106, and caesium-137 through adjusting physiological flux rates, in equilibrium with the changing levels of radionuclides in the environmental matrices, but not of the activation product nuclide cobalt-60.

The function of hemoglobin in the arcid clam Noetia ponderosa—I oxygenation in vitro and in vivo

Abstract 1. The mol. wt of the hemoglobin of the arcid clam Noetia ponderosa estimated on a calibrated gel column is 34,500, suggesting a dimeric molecule. 2. The oxygen affinity (P 50 ) of hemoglobin in intact cells in the pH range 6·6–7·4 is 5·30 mm Hg at 10°C and 6·25 mm Hg at 23°C. 3. Two phases of oxygen equilibrium curves can be detected, each with a significantly different Hill coefficient ( n ). The slope of the curve at lower oxygenation levels is slightly less than that at higher oxygenation levels. 4. Total oxygen content and PO 2 of the blood of animals exposed to various oxygen conditions indicate that substantial changes in oxygenation occur at environmental PO 2 's above 70 mm Hg, which are typical of the habitat. The measurements indicate that the blood from clams exposed to very low oxygen levels still contains detectable amounts of oxygen.

Effect of ionizing radiation on haemoglobin of marine lamellibranchs

The effect of ionizing radiation on the iron-linked protein (haemoglobin) of the marine lamellibranchs Anadara granosa (Linn.) and Cardita antiquata (Lam.) from Bombay waters, India is discussed. Purified haemoglobin solutions were exposed to a 60Cobalt source delivering a dose of about 4,600 rad/min, at the sample irradiation point. Radiation damage or degradation was measured spectrophometrically by studying changes in the absorption spectra following irradiation in the presence and absence of oxygen. Exposure to ionizing radiation in general caused a decrease in absorption of both haemoglobins, irrespective of location, viz extra-versus intracellular, at Soret (412 mμ), β (540 to 42 mμ) and α (574 to 76 mμ) peaks, and an increase in absorption at 510 and 630 mμ. Upon exposure to a higher dose, O2Hb of C. antiquata showed an increase in absorption at the protein peak (280 mμ); exposure to a lower dose, however, resulted in decreased absorption. Furthermore, the changes following irradiation were dependent upon the initial state of the pigment. Oxyhaemoglobin, when exposed to radiation, oxidized to hemiglobin, and hemiglobin reduced to oxyhaemoglobin. The extracellular haemoglobin of high molecular weight (3x106) of the false cockle C. antiquata was found to be extremely radio-resistant, whereas intracellular haemoglobin of low molecular weight (74,000) of the arcid clam A. granosa was highly radiosensitive, since it could not be exposed to doses exceeding 18,000 r.

Haemoglobin Variation in Anadara trapezia

THE most common respiratory pigment among the molluscs is haemocyanin, which is widely distributed among the gastropods, cephalopods and amphineurans. Most bivalves lack a respiratory pigment, but, if present, the pigment is haemoglobin, which can be found in muscle tissue, in haemocoelic erythrocytes or dissolved in the haemolymph1. In Anadara trapezia (Deshayes) (Mollusca, Bivalvia), an arcid clam, haemoglobin is present in haemocoelic erythrocytes.