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PubMed2005

Evolution of a unique mitotype-specific protein-coding extension of the cytochrome c oxidase II gene in freshwater mussels (Bivalvia: Unionoida).

A unique mode of mitochondrial DNA inheritance, designated doubly-uniparental inheritance (DUI), occurs in three bivalve subclasses (Pteriomorpha: Mytiloida, Palaeoheterodonta: Unionoida, Heterodonta: Veneroida), indicating that DUI may be a widespread phenomenon among bivalves. In mytiloids, breakdown of this pattern of inheritance (gender-switching) is observed in natural populations and in a phylogenetic context. In contrast, gender-switching has not occurred during the evolutionary history of unionoids. Here we present sequences for the male (M) and female (F) mitotypes from an additional 8 species of Unionoida. Consistent with previous observations, the M and F mitotypes of all species form reciprocally monophyletic clades supporting the hypothesis of taxon-specific rates of gender-switching. Coinciding with the absence of gender-switching is an approximately 185 codon extension of the cytochrome c oxidase II (MTCO2) locus in the male genome. The extension is present in all 12 unionoid species examined, including a representative of the family Margaritiferidae, indicating that this protein-coding polymorphism originated > or = 200 MYBP: . Although the extension is well conserved in length among 11 of the 12 species, one taxon has a significantly shortened extension. Lastly, examination of the rates and patterns of substitution indicate that the extension is evolving under relaxed purging selection, a pattern inconsistent with the conserved nature of MTCO2 or any cytochrome c oxidase locus.

Amino Acid Sequence↗
PubMed2004

Evaluation of lethality and genotoxicity in the freshwater mussel Utterbackia imbecillis (Bivalvia: Unionidae) exposed singly and in combination to chemicals used in lawn care.

Many chemicals, including fertilizers, herbicides, and insecticides, are routinely applied to turf in the care and maintenance of lawns. These chemicals have the potential to leach into nearby surface waters and adversely affect aquatic biota. In this study, we evaluated the lethal and genotoxic effects of chemicals used in lawn care on an early life stage of freshwater mussels (Utterbackia imbecillis). The chemicals tested were copper and commercial formulations of atrazine, glyphosate, carbaryl, and diazinon. Mussel glochidia were exposed to chemicals singly or in combination (equitoxic and environmentally realistic mixtures) for 24 h and toxic interactions were evaluated with Marking's additive index. Genotoxicity was quantified with the alkaline single-cell gel electrophoresis assay (Comet assay). In acute tests, copper was the most toxic of all chemicals evaluated (LC50 = 37.4 microg/L) and carbaryl was the most toxic of all pesticides evaluated (LC50 = 7.9 mg/L). In comparison to other aquatic organisms commonly used in toxicity tests (e.g., amphipods, cladocerans, and chironomids), mussel glochidia were as or more sensitive to the chemicals evaluated with the exception of diazinon, where mussels were observed to be less sensitive. The combined toxicity of equitoxic and environmentally realistic mixtures to mussels was additive. Genotoxic responses were observed in mussels exposed to copper, atrazine and diazinon at levels below their respective no-observed-effect concentrations. Together, these data indicate that freshwater mussels are among the most sensitive aquatic organisms tested for some chemicals commonly used in lawn care and that DNA damage may be useful as a screening tool to evaluate potential sublethal effects of lawn care products on non-target aquatic organisms.

Animals↗
PubMed2006

A new Paravortex (Platyhelminthes, Dalyellioida) endoparasite of Mesodesma mactroides (Bivalvia, Mesodesmatidae) from Uruguay.

Many species of turbellarians (Platyhelminthes) are known to live associated with other organisms, especially invertebrates, as commensals or parasites. The family Graffillidae (Rhabdocoela) includes two genera that parasitize mollusks, Graffílla and Paravortex. Within the latter genus, six species were described as associated with mollusks. In other instances, unnamed Paravortex species were mentioned as parasites of other bivalves and of the body surface of fishes. In the present work, a new Paravortex species that was found in the intestine of Mesodesma mactroides from the Atlantic coast of Uruguay is described. In addition, a bibliographical revision of the known Paravortex species with their respective hosts, location, and distribution is made. Paravortex nicolli, described by Szidat for the Argentinean coast, is mentioned for the first time after the original description, and the authorship and date of description of Paravortex tapetis Noury-Sraïri 1989 are elucidated.

Animals↗
PubMed1996

Chaetogaster limnaei (annelida: oligochaeta) as a parasite of the zebra mussel Dreissena polymorpha, and the quagga mussel Dreissena bugensis (mollusca: bivalvia).

Dreissenid mussels, Dreissena polymorpha and D. bugensis, were found to be infected by the naidid oligochaete Chaetogaster limnaei at four sites in the St. Lawrence River. This is the first report of this species infecting dreissenids anywhere in the world. Most worms inhabited the mantle cavity, where they caused erosion of the mantle and gill epithelia as determined by histopathological examination. Others penetrated various tissues; one had invaded the ovary and was feeding on oocytes and ovarian tissues. Of 606 mussels examined, 166 (27.4%) harbored at least 1 C. limnaei. The prevalence varied between 1% and 80%, depending on the collection site and date. The worms were slightly but significantly more prevalent in D. bugensis than in D. polymorpha. The intensity ranged from 1 to 18 worms per infected host. Variations in prevalence and intensity were not related to the size or sex of the host, but the data did suggest some seasonality.

Animals↗
PubMed2006

Bacterial endosymbioses in Solemya (Mollusca: Bivalvia)--model systems for studies of symbiont-host adaptation.

Endosymbioses between chemosynthetic bacteria and marine invertebrates are remarkable biological adaptations to life in sulfide-rich environments. In these mutualistic associations, sulfur-oxidizing chemoautotrophic bacteria living directly within host cells both aid in the detoxification of toxic sulfide and fix carbon to support the metabolic needs of the host. Though best described for deep-sea vents and cold seeps, these symbioses are ubiquitous in shallow-water reducing environments. Indeed, considerable insight into sulfur-oxidizing endosymbioses in general comes from detailed studies of shallow-water protobranch clams in the genus Solemya. This review highlights the impressive body of work characterizing bacterial symbiosis in Solemya species, all of which are presumed to harbor endosymbionts. In particular, studies of the coastal Atlantic species Solemya velum and its larger Pacific congener Solemya reidi are the foundation for our understanding of the metabolism and physiology of marine bivalve symbioses, which are now known to occur in five families. Solemya velum, in particular, is an excellent model organism for symbiosis research. This clam can be collected easily from coastal eelgrass beds and maintained in laboratory aquaria for extended periods. In addition, the genome of the S. velum symbiont is currently being sequenced. The integration of genomic data with additional experimental analyses will help reveal the molecular basis of the symbiont-host interaction in Solemya, thereby complementing the wide array of research programs aimed at better understanding the diverse relationships between bacterial and eukaryotic cells.

Adaptation, Physiological↗
PubMed2005

Surface spreading of synaptonemal complexes in the clam Dosinia exoleta (Mollusca, Bivalvia).

There are only a few reports on the chromosomal location of DNA sequences in bivalve species, none of them using meiotic chromosomes. Mitotic chromosomes of the clam Dosinia exoleta were analysed by means of Giemsa, silver and fluorochrome staining and fluorescent in situ hybridization (FISH) with 18S + 28S rDNA and telomeric probes. A technique for surface spreading of synaptonemal complexes (SCs) of Dosinia exoleta was developed for the first time in a bivalve species. Silver and DAPI/PI staining and SC-FISH were also applied to the study of the meiotic chromosomes of this clam. The diploid chromosome number in this species is 38 and the karyotype is composed of 11 pairs of metacentric and eight pairs of submetacentric chromosomes. 18S + 28S rDNA clusters map to the subtelomeric region of the short arm of one metacentric chromosome pair whereas telomeric signals appear at both ends of every chromosome.

Animals↗
PubMed1990

Number and types of hemocytes in Sunetta scripta and Villorita cyprinoides var. cochinensis (Bivalvia), and leukocytosis subsequent to bacterial challenge.

The number and types of hemocytes in four size groups of the clam species Sunetta scripta and Villorita cyprinoides var. cochinensis, and leukocytosis in the 38- to 40-mm-size-group clams of both species subsequent to challenge with Vibrio alginolyticus at a concentration of 1 x 10(8) cells/0.02 ml of sterile 2% saline was investigated. In S. scripta, the mean total hemocyte count in the 42- to 44-mm size group was significantly lower than that of the three other size groups but there was no significant variation in total cell counts in the four size groups of V. cyprinoides var. cochinensis. Only two types of hemocytes, granulocytes and agranulocytes, occur and the percentage of agranulocytes was roughly half of that of granulocytes. The data on the effects of sham injection and Vibrio injection suggest that there is significant leukocytosis early in both clam species as a result of sham injection; the bacterial challenge produces significant leukocytosis in V. cyprinoides var. cochinensis both early (6 and 12 hr) and later (48,96 and 120 hr), but only at 48 hr in S. scripta; and in both clam species there is significant increase in total cell counts in Vibrio-injected ones than in untampered controls at various time intervals.

Animals↗
PubMed1982

The molluscan neuropeptide FMRFamide: calcium-dependent release and blood levels in Macrocallista (Bivalvia).

Ganglia dissected from the marine bivalve, Macrocallista nimbosa, were pooled and incubated in a series of seawater media designed to promote or inhibit the release of the neuropeptide Phe-Met-Arg-Phe-NH2 (FMRFamide). The amount of peptide released, determined by radioimmunoassay (RIA), was enhanced two to seven times when the medium contained high potassium. FMRFamide released was reversibly inhibited when calcium was omitted from the high potassium medium. Immunoreactive FMRFamide was found in Macrocallista hemolymph in a concentration (10(-9)M) that excites isolated Macrocallista ventricles. Although the in vivo release sites have not yet been localized, these findings already show that the endogenous neuropeptide FMRFamide can be characterized as a neurohormone or as a neurotransmitter in Macrocallista.

Animals↗
PubMed1989

High affinity dopamine binding to mouse thymocytes and Mytilus edulis (Bivalvia) hemocytes.

The present report demonstrates that both mouse thymocytes and Mytilus edulis hemocytes contain a novel type of dopamine receptor. Scatchard analysis of these data revealed a single class of high-affinity binding sites with an affinity constant (Kd) of 6.6 nM, and a binding site density (Bmax) of 141 pmol/g protein in mice and a Kd of 7.6 nM and a Bmax of 66 pmol/g protein for the hemocytes. In older Mytilus the Kd was the same; however, there was a significant decrease in the Bmax (48 pmol/g protein; P greater than 0.05). Age changes were not evident for the mouse cells. The ability of a variety of other related drugs to displace specifically bound dopamine was investigated. The catecholamines (norepinephrine greater than epinephrine) and the dopamine agonist epinine were the most potent of the ligands tested and apomorphine and butaclamol were of moderate potency. This may serve to indicate that this dopamine binding site may be very different from the known D1 and D2 sites. This particular site may represent a third type of dopaminergic receptor. Furthermore, the hemocytes of Mytilus appear to be of two major types, namely, cells with a granular cytoplasm and those whose cytoplasm is agranular. Histofluorescence studies on the hemocytes reveal that a subpopulation of the granule-containing cells appears to contain serotonin.

Animals↗
PubMed1994

Use of monoclonal antibodies for identification of growth-controlling neuropeptides in the mussel Mytilus edulis (Mollusca:Bivalvia).

Monoclonal antibodies were developed against cerebral ganglia (CG) of the mussel Mytilus edulis by the immunization of mice with unpurified homogenates of these organs. The screening protocol of hybridoma was based upon immunohistological observations of cytocentrifugated ganglia cells. A panel of 29 monoclonal antibodies (MABs) specific of CG epitopes was harvested and subsequently used for the immunocytochemical study of CG cells. Several subpopulations of ganglia cells were specifically revealed by MABs. Identification of epitopes involved in growth control was approached via the application of a bioassay allowing the assessment of protein synthesis stimulation. MAB 42 and 46 affected amino acid incorporation induced by CG extract. These results lead to the conclusion that the epitopes recognised by these antibodies are involved in growth control. An immunoenzymatic assay was performed with CG extracts for quantitative analyses of epitopes.

Amino Acids↗
PubMed1990

A comparative study of alkaline phosphatases among human placenta, bovine milk, hepatopancreases of shrimp Penaeus monodon (Crustacea: Decapoda) and clam Meretrix lusoria (Bivalvia: Veneidae): to obtain an alkaline phosphatase with improved characteristics as a reporter.

1. Alkaline phosphatases were purified from human placenta, bovine milk, shrimp and clam with a final spec. act. of 67,000, 32,000, 22,000 and 15,000 U/mg of protein respectively. 2. The alkaline phosphatase from Meretrix lusoria is unique with its thermostability at 65 degrees C for 30 min; whereas the remaining enzymes studied, including the human placental alkaline phosphatase, are inactivated and have negligible activities. 3. The alkaline phosphatase from Penaeus monodon can be differentiated by its pH optimum at 9.0; the remaining enzymes studied have their optimal pH at 10.0. 4. The alkaline phosphatases from shrimp and clam are proposed to be applied as "reporters" in the study of mammalian cells.

Alkaline Phosphatase↗
PubMed1993

Bioaccumulation and distribution of mercury in the hard clam, Meretrix lusoria (Bivalvia: Veneidae).

1. Total mercury concentration in whole soft parts of the hard clam, Meretrix lusoria, increased significantly with increasing exposure concentrations. It reached 4.247-7.084 and 9.956-13.643 micrograms T-Hg/g dry weight in 5 and 50 micrograms/l Hg mercury solution, respectively, against a background level of 1.824-0.577 micrograms T-Hg/g dry weight. 2. Bioconcentration factor was 4-6 times higher in 5 micrograms/l Hg than that of 50 micrograms/l Hg. An estimated biological half-life of 20.2 and 18.4 days in 5 and 50 micrograms/l Hg, respectively, were also obtained after 14 days uptake. 3. Accumulation of mercury in tissues of the clams was greater in the gills and viscera than in the mantle, adductor, foot, or hemolymph. 4. The amount of mercury present in the gills is related to be a linear relationship with the mercury content of viscera.

Animals↗
PubMed2006

Heart rate and burrowing behavior in the mussel Anodontites trapesialis (Bivalvia: Mycetopodidae) from lotic and lentic sites.

Burrowing rate index, diurnal variations in heart rate and heart rate alterations associated with behavioral patterns were studied in Anodontites trapesialis from the Pardo River (PR) and the Galo Bravo Dam (GBD). In both populations the pattern immobile/buried was associated with the lowest (basal) values of heart rate, which increase significantly during burrowing remaining steady in its phases I and II and at the first 10 min of phase III. A decrease in the frequency of digging cycles was significantly correlated with the gradual decrease in heart rate during the phase III of burrowing. Nevertheless, mussels from PR burrow faster than those of GBD and possess higher values of basal heart rate which present a diurnal variation with the higher values occurring during the light phase. Yet, the basal heart rate in mussels from GBD remained stable over the course of 24 h. Also, the increase in heart rate associated with burrowing was higher in mussels from PR and compatible with the highest burrowing index previously detected. Since mussels possess a plastic phenotype it is probable that the cited functional differences are phenotypic adaptations, but we cannot exclude the possibility of these adaptations being genotypic in nature.

Animals↗
PubMed2006

Immunological and biochemical responses in Mya arenaria (Mollusca Bivalvia) exposed in vivo to estradiol-17beta.

Soft-shell clams Mya arenaria were injected with 10, 20 or 40 nmol of estradiol 17beta (E2). We observed a significant inhibiting effect of E2 on phagocytic activity of hemocytes from clams exposed to 10 and 20 nmol. A dose-response increase of the glycogen phosphorylase in the gonad tended to show a remobilisation of glycogen reserves involved in vitellogenesis although the exposure time must have been too short to observe a decrease in glycogen reserves or an increase in RNA concentration. Both results corroborate those of other studies about estrogen involvement in controlling immune capacity and energy metabolism related to vitellogenesis in bivalves. We can assume that immune parameters should now be taken into consideration in assessing endocrine disruption in bivalves. Nevertheless further studies are needed to understand the controlling pathways of E2 with a special regard on its interactions with other effectors involved in bivalve immunity and reproduction as well.

Animals↗
PubMed2004

Do organisms living around hydrothermal vent sites contain specific metallothioneins? The case of the genus Bathymodiolus (Bivalvia, Mytilidae).

The unusual characteristics of the hydrothermal vent environment (high pressure and metal concentrations, low pH, etc.) leads us to wonder how species living in this particular biotope have adjusted to these severe living conditions. To investigate the consequences of high metal concentrations, filter-feeding organisms are commonly used in ecotoxicological studies. Metallothioneins (MTs) are proteins conserved throughout the animal kingdom and involved in intracellular metal regulation. Therefore, we tried here to find out whether the metallothioneins of hydrothermal bivalves are different from those of coastal bivalves. The characterization of DNA sequences coding MTs from some of the most common hydrothermal bivalves, belonging to the genus Bathymodiolus (Mytilidae) was performed by reverse transcription polymerase chain reactions (RT-PCRs). The complementary DNAs (cDNAs) of MT-10 and MT-20 isoforms were obtained for the Atlantic and Pacific hydrothermal mussels (Bathymodiolus azoricus and Bathymodiolus thermophilus). The MT-10 transcripts were 222 nucleotides long and the MT-20 transcripts, 207 nucleotides. The polymorphism of the MT cDNAs in these two hydrothermal species is discussed. The comparison between metallothionein cDNA sequences of the Mytilus and the Bathymodiolus genera shows strong homologies among metallothioneins of coastal and hydrothermal mussels.

Adaptation, Physiological↗
PubMed2006

Analysis of EST and lectin expressions in hemocytes of Manila clams (Ruditapes philippinarum) (Bivalvia: Mollusca) infected with Perkinsus olseni.

The hemocytes of invertebrates play key roles in both cellular and humoral immune reactions by phagocytosis or delivering immune factors such as lectin and anti-microbial peptides. Bacterial infection causes changes in components such as lectins, anti-bacterial peptides, and lysosomal enzymes of plasma or hemolymph in molluscs. Previously, we found that infection with the protozoan parasite, Perkinsus, increases lectin synthesis in hemocytes. In order to investigate the patterns of genes expressed in Manila clams (Ruditapes philippinarum) infected with the protozoan parasite Perkinsus olseni, we constructed a cDNA library and sequenced 1850 clones (expressed sequence tags). A total of 79 ESTs, were related to 29 functional immune genes such as C-type lectin, lysozyme, and cystatin B, in Manila clams. Lectins were the largest group of immune-function ESTs found in our Manila clams library. Among 7 lectin clones, two full length cDNAs of lectins were cloned. MCL-3, which is a simple C-type lectin composed of 151 amino acids, has a relatively short signal sequence of 17aa and single carbohydrate-recognition domain (CRD) of approximately 130 residues. It is highly homologous to eel C-type lectin. The sequence of mc-sialic acid-binding lectin consists of 168 amino acid residues with molecular weight of 19.2 and shows high homology to sialic acid-binding lectin from the snail, Cepaea hortensis. The expression of 7 different lectins in hemocytes was analyzed by RT-PCR using gene-specific primers. Hemocytes from Perkinsus-infected clam expressed different sets of lectins than with Vibrio infection. These results demonstrate that several lectins are involved in Manila clam innate immunity and different challenges induce expression of different lectins.

Amino Acid Sequence↗
PubMed2005

Use of sodium dodecyl sulfate and zinc sulfate as reference substances for toxicity tests with the mussel Perna perna (Linnaeus, 1758) (Mollusca: Bivalvia).

Effects of anthropogenic pollution have been observed at different trophic levels in the oceans, and toxicity tests constitute one way of monitoring these alterations. The present assay proposes the use of two reference substances, sodium dodecyl sulfate (SDS) and zinc sulfate, for Perna perna larvae. This common mussel on the Brazilian coast is used as a bioindicator and is of economic interest. The chronic static embryo-larval test of short duration (48 h) was employed to determine the NOEC, LOEC, and IC50 for SDS and zinc sulfate, as well as the coefficient of variation. Salinity, pH and un-ionized ammonia (NH3) and dissolved oxygen (DO) concentrations were measured to monitor water quality. The results demonstrated that the main alterations in veliger larvae are the development of only one shell, protruded mantle, malformed shell, formation of only part of a valve, clipped edges, uneven sizes and presence of a concave or convex hinge. NOEC values were lower than 0.25 mg L(-1) for zinc sulfate and 0.68 mg L(-1) for SDS. The coefficient of variation was 17.63% and 2.50% for zinc sulfate and SDS, respectively.

Animals↗
PubMed2006

Ultrastructural study of neoplastic cells in Macoma balthica (Bivalvia) from the Gulf of Gdansk (Poland).

In the Baltic clam Macoma balthica from the Gulf of Gdansk, neoplasia is considered as a serious epizootic linked to bad environmental conditions and high levels of pollutants. Previous research was focused on the diagnosis, prevalence, seasonality and histopathological characteristic of the cancer. This study is focused on electron microscopy analyses describing cell ultrastructure abnormalities related to neoplasia. Examinations using the electron microscopy highlighted changes confined to anatomic ultrastructures, shapes and functions of neoplastic cells. The lobulated appearance of the nucleus, changes in cellular matrix and the occurrence of large granular cells with hyperchromatic nuclei, atypical Golgi structures and deterioration of rough endoplasmic reticulum manifested the disease. The presence of atypical mitochondria, free ribosomes and hypertrophic nuclei suggests the adaptation of neoplastic cells to increased mitotic activity, while the observed modification of cellular membranes may reflect functional changes connected to increased pinocytotic activity or intercellular transport. The cancer cells were found to appear in two types, abnormal round-shaped cells and spindle-shaped cells, both with increased frequencies of cell division. Round-shaped cells typical for disseminated neoplasia were observed in all affected bivalves, in a few cases co-occurring with abnormal spherical cells. Spindle-shaped cells containing some intracytoplasmic filaments, and with a tendency of the nuclei to be orientated as in a palisade were interpreted as myofibroblasts-like cells and were observed in five out of eleven clams diagnosed as neoplastic. This finding represents the first demonstration of phenotypic differences in the cell types co-occurring in animals diagnosed as neoplastic and by that suggests coexistence of two types of bivalve cancer, disseminated neoplasia and probable fibrosarcoma.

Animals↗