Gonad morphology and reproductive cycle of the mangrove oyster Crassostrea brasiliana (Lamarck, 1819) in the baía de Guaratuba,Paraná, Brazil

This study aimed to describe the gonadal histology and the reproductive cycle of Crassostrea brasiliana in the mangroves of Guaratuba Bay in southern Brazil. Adults were collected monthly from January 2010 to April 2011 from three sampling sites in intertidal oyster beds. The animals were evaluated using biometric and histological analyses of the gonads. The gonadal tissue samples were processed according to the standard histological procedures, and permanent slides were prepared using Harris' haematoxylin and eosin. The oysters were identified at the species level using a molecular protocol. Females (69%) predominated over males (26%), with 4% indeterminate and 1% hermaphroditic. Mature females were more prevalent in February, March and December 2010 and in March 2011. Mature males were more prevalent in February and April 2010 and in March 2011. The presence of hermaphroditic individuals was sporadic, and oysters in immature stages or sexual repose were observed in only a few collections between the months of May and October 2010. The reproduction of C. brasiliana in Guaratuba Bay occurs intermittently, but with greater intensity during the summer, with a larger number of females produced.     

Fine structure of the spermatozoa of Crassostrea gigas (Mollusca,Bivalvia)

We describe sperm ultrastructure and acrosome differentiation during spermiogenesis in Crassostrea gigas (Mollusca Bivalvia). The sperm cell is a uniflagellated cell of the primitive type. The head region contains a rounded or conical nucleus surmounted by small acrosome. This organelle consists of a membrane-bound acrosomal granule, the contents of which have a homogeneous density, except in the anterior region, which is positive for PTA. The acrosome also surrounds the perforatorium, which includes oriented fibrillar elements: this is the axial body. The middle piece contains four mitochondria encircling two perpendicular centrioles. The distal centriole is provided with a system of mechanical fixation to the plasma membrane, consisting of nine fibers in radial arrangement. The tail flagellum, about 50 m?m long, contains the usual microtubular axoneme. © 1993 Wiley-Liss, Inc.     

Biochemical and energy dynamics throughout the reproductive cycle of the striped venus Chamelea gallina (Mollusca,Bivalvia)

The striped venus Chamelea gallina is an important commercial bivalve species in Europe. However, large inter-annual fluctuations in stock abundance and periodic recruitment failure threaten the biological and economic sustainability of this fishery. This study aimed to improve the knowledge of the reproductive cycle and reproductive strategies of this species from the Algarve coast (southern Portugal) in order to contribute to the establishment of management measures and to assess its potential for aquaculture. The reproductive cycle of C. gallina followed a seasonal cycle, significantly influenced by sea surface temperature and food availability. Gametogenesis took place in winter, coinciding with the phytoplankton bloom. Spawning occurred during summer, followed by a short period of sexual inactivity in autumn. Condition index did not reflect the reproductive cycle of C. gallina and generally, followed the same trend of chlorophyll a. Glycogen was positively correlated with gonadal index and chlorophyll a. High total lipid values were recorded throughout gonad ripeness and spawning, but decreased at the end of the spawning and in the rest period. The extended spawning period of C. gallina will allow larvae to be obtained for much of the year by artificial spawning of wild broodstock.     

Reproductive cycle of the mangrove oyster Crassostrea rhizophorae (Bivalvia: Ostreidae) in Camamu Bay, Bahia, Brasil

The mangrove oyster Crassostrea rhizophorae is important fishery resource along the entire Brasilian coast with excellent potential for marine culture. The purpose of this paper was to examine the reproductive characteristics of the oyster of the Maraú river estuary in Camamu Bay, Bahia, Brasil. The samples were collected monthly, from September 2006 to August 2007, at two points (I and II) in Camamu Bay. At each site 20 oysters were collected for histological analysis, fixed in Davidson's solution, embedded in paraffin, dehydrated in an ethanol series, sectioned (7 microm thick) and stained with Harris hematoxylin and Eosin (HE). Additionally, 30 oysters were sampled, at each point, for a condition index analysis. The water temperature ranged from 23.5 degrees C to 30 degrees C and the salinity from 15 to 25 ups at Point I (Maraú) and from 25 to 35 at Point II (Tanque Island). The oyster's height ranged from 30 to 92 mm at Point I and from 27 to 102 mm at Point II, with an average of 49.0 mm +/- 9.1 (n = 230) and 49.9 mm +/- 9.9 (n = 237), respectively. Among the sampled oysters at Point I, 59.1% were females, 31.3% males, 1.3% hermaphrodites and 8.2% of the oysters of undetermined sex. At Point II, 66.2% were females, 30.4% males, 0.8% hermaphrodites and 2.5% (n = 237) of undetermined sex. The gonadic stage analysis indicated that the reproduction period of the C. rhizophorae in the Maraú Peninsula was continuous all year, without any regressive phase. The condition index (R) ranged from 8.0% to 17.7%. The peak periods of R coincided with the expressive oyster's percentage in the maturation and liberation gametic stages. The results of these findings will contribute information for the oyster spat collection and to the process installation of the oyster culture in Camamu Bay.     

Differences in the rDNA-bearing chromosome divide the Asian-Pacific and Atlantic species of Crassostrea (Bivalvia, Mollusca)

Karyotype and chromosomal location of the major ribosomal RNA genes (rDNA) were studied using fluorescence in situ hybridization (FISH) in five species of CRASSOSTREA: three Asian-Pacific species (C. gigas, C. plicatula, and C. ariakensis) and two Atlantic species (C. virginica and C. rhizophorae). FISH probes were made by PCR amplification of the intergenic transcribed spacer between the 18S and 5.8S rRNA genes, and labeled with digoxigenin-11-dUTP. All five species had a haploid number of 10 chromosomes. The Atlantic species had 1-2 submetacentric chromosomes, while the three Pacific species had none. FISH with metaphase chromosomes detected a single telomeric locus for rDNA in all five species without any variation. In all three Pacific species, rDNA was located on the long arm of Chromosome 10 (10q)--the smallest chromosome. In the two Atlantic species, rDNA was located on the short arm of Chromosome 2 (2p)--the second longest chromosome. A review of other studies reveals the same distribution of NOR sites (putative rDNA loci) in three other species: on 10q in C. sikamea and C. angulata from the Pacific Ocean and on 2p in C. gasar from the western Atlantic. All data support the conclusion that differences in size and shape of the rDNA-bearing chromosome represent a major divide between Asian-Pacific and Atlantic species of CRASSOSTREA: This finding suggests that chromosomal divergence can occur under seemingly conserved karyotypes and may play a role in reproductive isolation and speciation.     

Reconstructing the Anomalodesmata (Mollusca: Bivalvia): morphology and molecules

The extant anomalodesmatan bivalves have always proved rather enigmatic and difficult to interpret, both in terms of their relationships to other bivalve taxa and the interrelationships of the constituent families. These difficulties stem from their diverse and often highly specialized life habits which have resulted in a wide array of disparate morphologies, and also from the fact that many are extremely rare. Classifications based on morphological characters have been dogged by fears that convergent and parallel evolution has masked phylogenetic signals. Molecular surveys of members of 12 of the 15 constituent families, using the 18S rRNA gene, have revealed that anomalodesmatans are robustly monophyletic and lie within the basal heterodonts. The Anomalodesmata should no longer be regarded as a subclass, but as a part of the Heterodonta. Here we present an enhanced analysis of 32 anomalodesmatan species (representatives of 12 families). Our results, subjected to Maximum Parsimony, Maximum Likelihood and Bayesian analyses, challenge our understanding of the internal relationships within the Anomalodesmata. In particular they indicate the need for a re-distribution of the families traditionally placed in the Thracioidea and Pandoroidea into a 'thraciid' lineage (Thraciidae + Cleidothaeridae + Myochamidae) and a 'lyonsiid' lineage (polyphyletic Lyonsiidae + Clavagellidae + Laternulidae + Pandoridae). The endolithic Clavagella and endobenthic Brechites and Penicillus form a robust clade. The hypothesis that the carnivorous septibranchs are monophyletic can, thus far, be neither supported nor rejected. Mapping critical morphological characters onto our molecular results provides evidence of multiple loss of some characters (e.g. prismato-nacreous shell microstructure and shell spicules) and also multiple gain of others (e.g. chondrophores).  © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society , 2006, 148 , 395–420.     

Fine structure of the early stages of spermatogenesis in the Pacific oyster, Crassostrea gigas (Mollusca, Bivalvia)

The aim of this study is to describe the early stages of spermatogenesis of the Pacific oyster Crassostrea gigas using both light and electron microscopy. The gonad is formed by gonadal tubules invaginated in a connective tissue constituting a storage tissue. Myoepithelial cells surround each gonadal tubule and are associated with an acellular matrix delimiting the outer part of the tubule, the inner part is composed by intragonadal somatic cells associated with germinal lineage. Two types of spermatogonia are identified, where type I spermatogonia (Spg I) are large, scarce and pale cells leaned against the base of the tubule (nuclear diameter: 5.5+/-0.5 microm). Type II spermatogonia (Spg II) are clustered and dark cells which appear smaller than type I (nuclear diameter: 4.3+/-0.3 microm). The aspect of nuage-like material in cytoplasm is described from pale spermatogonia to primary spermatocytes (nuclear diameter: pachytene 3.6+/-0.3 microm, diplotene 3.4+/-0.3 microm), while no structure related to a chromatoid body was observed in oyster spermatocytes and spermatids.     

Annual reproductive cycle and condition index of Mactra corallina (Mollusca: Bivalvia) from the north coast of Tunisia

This work aims to examine the reproductive cycle of the bivalve Mactra corallina inhabiting the Gulf of Tunis (Tunisia). Histological analysis indicated that the reproductive cycle is composed of six stages and was synchronous between sexes. Gonadal development began in January. From April to July, an increase in temperature (from 16 to 28°C) and a rise in chlorophyll a (from 3.25 to 6.35 mg/L) marked the period of sexual maturity in both sexes, which was also confirmed by an elevation in condition index (CI). Partial spawning started in April for males and in May for females, peaking in late summer indicating a prolonged reproductive period. The reproductive cycle of M. corallina finished with an inactive stage during autumn. From the total number of animals used in this study, we identified 104 females (52%) and 96 males (48%) corresponding to a sex ratio 1:0.92 that did not differ significantly from a 1:1 ratio.     

Phylogeny and spatio-temporal distribution of European Pectinidae (Mollusca: Bivalvia)

Phylogenetic relationships within Pectinidae (Bivalvia, Pteriomorphia) have been investigated primarily for Pacific and Western Atlantic or commercially valuable taxa. Most molecular phylogenetic studies have revealed monophyly of pectinid bivalves but interrelationships of the different clades are still inconsistent. However, non-commercial European Pectinidae has mostly been neglected in earlier investigations and therefore the evolution and radiation of the European Pectinidae is poorly understood. Since the fossil record of this group is well investigated, the evolutionary age of phylogenetic diversification and radiation events within this group can be dated. Thus, the connection of geological and climatic changes to radiation events within this group can be assessed. We investigated the phylogenetic relationships within European Pectinidae using mitochondrial (12S and 16S) and nuclear (18S, 28S and H3) gene markers and performed relaxed molecular clock approaches to gain information on the evolutionary age and the connection between Cenozoic climatic changes and diversification within this group. The results show concordance of radiation events with the Middle Miocene cooling event and the following climatic period with slowly decreasing temperatures. However, geological changes such as the uplift of the Gomphotherium Landbridge or the closure and re-opening of the Strait of Gibraltar also had great impact on diversification and distribution patterns within European Pectinidae.     

Fine structural study of the spermatogenic cycle in Pitar rudis and Chamelea gallina (Mollusca,Bivalvia, Veneridae)

A comparative ultrastructural study of spermatogenesis was performed in the bivalve molluscs Pitar rudis and Chamelea gallina (Veneridae) from Turkey. Sertoli cells appeared to be rich in glycogen, lipid droplets and germ-cell phagolysosomes. Premeiotic cells exhibited nuage and a flagellum, with the Golgi complex and the rough endoplasmic reticulum originating proacrosomal vesicles during the pachytene stage. In round spermatids, the acrosomal vesicle migrated linked to the plasma membrane. In P. rudis, the acrosomal vesicle base formed a thin expansion that attached to the nuclear apex and was associated with development of the perforatorium. The cap-shaped acrosomal vesicle then differentiated into external and internal regions, and also into a small apical light region, although some cells exhibited an apical extension of the external component. On the contrary, two lateroapical light pouches developed in C. gallina. During spermiogenesis, chromatin became fibrillar and then condensed while the nucleus turned conical shaped in P. rudis or slightly curved in C. gallina. In P. rudis, the midpiece contained glycogen and four mitochondria, although five mitochondria were sometimes observed, whereas in C. gallina the midpiece contained four mitochondria. Comparison with other members of Veneroida shows a common ectaquasperm type, but novel findings in acrosome biogenesis.     

Human-facilitated dispersal of the Gulf toadfish Opsanus beta (Goode & Bean, 1880) in the Guaratuba Bay,south-eastern Brazil

Opsanus beta is endemic to the Gulf of Mexico and has recently been introduced to the Brazilian coast; probably the introduction is via ballast water and/or oil rigs. In this study, the presence of the species is recorded for the first time in Guaratuba Bay, on the southern coast of Brazil. In this region there are no port terminals, which suggests that O. beta used a different mode of human-facilitated transport to colonize Guaratuba Bay.     

Numerical quantification of Perkinsus marinus in the American oyster Crassostrea virginica (Gmelin, 1791) (Mollusca: Bivalvia) by modern stereology

Species of Perkinsus are responsible for high mortalities of bivalve molluscs world-wide. Techniques to accurately estimate parasites in tissues are required to improve understanding of perkinsosis. This study quantifies the number and tissue distribution of Perkinsus marinus in Crassostrea virginica by modern stereology and immunohistochemistry. Mean total number of trophozoites were (mean +/- SE) 11.80 +/- 3.91 million and 11.55 +/- 3.88 million for the optical disector and optical fractionator methods, respectively. The mean empirical error between both stereological approaches was 3.8 +/- 1.0%. Trophozoites were detected intracellularly in the following tissues: intestine (30.1%), Leydig tissue (21.3%), hemocytes (14.9%), digestive gland (11.4%), gills (6.1%), connective tissues (5.7%), gonads (4.1%), palps (2.2%), muscle (1.9%), mantle connective (0.8%), pericardium (0.7%), mantle epithelium (0.1%), and heart (0.1%). The remaining 0.6% were found extracellularly. Percentages of trophozoite stages were (mean +/- SE): large, log-phase trophonts, i.e., signet rings, 97.0 +/- 1.2%; meronts, 2.0 +/- 0.9%; clusters of small, log-phase trophonts, i.e., merozoites, 1.0 +/- 0.5%. Levels of infection in hemocytes and Leydig tissue were representative of total parasite intensity. These techniques are a powerful tool to follow parasite distribution and invasion, and to further explore mechanisms of Perkinsus spp. pathogenesis in bivalves.     

Role of Unionidae and Sphaeriidae (Mollusca,Bivalvia) in the Eutrophic Lake Zbęchy and its Outflow

The abundance, biomass, respiration, and filtration rate of the Unionidae and Sphaeriidae and their significance to the phosphorus cycle was estimated in an eutrophic lake in Poland, situated in an agricultural area, and in its outflow canal. A decrease in Unionidae biomass was compensated for the Sphaeriidae in terms of both biomass and filtration rate. Depending on the developmental state of the Unionidae and Sphaeriidae populations, the volume of pumped water, the amount of removed and assimilated seston and the phosphorus excretion changed significantly.     

Diel variation of the RNA/DNA ratios in Crassostrea angulata (Lamarck) and Ruditapes decussatus (Linnaeus 1758) (Mollusca: Bivalvia)

The aim of this study was to investigate the effect of time of day on RNA/DNA ratios among fed and starved Crassostrea angulata and Ruditapes decussatus juveniles. Sampling to investigate the day and night condition of juveniles was carried out for 48 h. A highly sensitive method for nucleic acid quantification was applied to bivalves. The results suggest that there is some variation in nucleic acid quantities with the time of the day. For the two species analysed, the RNA/DNA ratio was particularly high during the night and was higher in the fed animals. The results seem to indicate that there is some endogenous rhythm in the production of RNA. If there are diel changes in RNA/DNA ratios, it follows that average RNA/DNA ratios can be unrepresentative if there is any day or night bias in sampling.     

Settlement and Survival of the Oyster Crassostrea virginica on Created Oyster Reef Habitats in Chesapeake Bay

Efforts to restore the Eastern oyster (Crassostrea virginica) reef habitats in Chesapeake Bay typically begin with the placement of hard substrata to form three‐dimensional mounds on the seabed to serve as a base for oyster recruitment and growth. A shortage of oyster shell for creating large‐scale reefs has led to widespread use of other materials such as Surf clamshell (Spisula solidissima), as a substitute for oyster shell. Oyster recruitment, survival, and growth were monitored on intertidal reefs constructed from oyster and Surf clamshell near Fisherman’s Island, Virginia, U.S.A. and on a subtidal Surf clamshell reef in York River, Virginia, U.S.A. At the intertidal reefs, oyster larvae settlement occurred at similar levels on both substrate types throughout the monitoring period but higher levels of post‐settlement mortality occurred on clamshell reefs. The oyster shell reef supported greater oyster growth and survival and offered the highest degree of structural complexity. On the subtidal clamshell reef, the quality of the substrate varied with reef elevation. Large shell fragments and intact valves were scattered around the reef base, whereas small, tightly packed shell fragments paved the crest and flank of the reef mound. Oysters were more abundant and larger at the base of this reef and less abundant and smaller on the reef crest. The availability of interstitial space and appropriate settlement surfaces is hypothesized to account for the observed differences in oyster abundance across the reef systems. Patterns observed emphasize the importance of appropriate substrate selection for restoration activities to enhance natural recovery where an underlying habitat structure is destroyed.     

Global diversity of freshwater mussels (Mollusca, Bivalvia) in freshwater

The term freshwater bivalve is very inclusive and not very informative. There are representatives of at least 19 families that have at least one representative living in freshwater. This suggests at least 14 different invasions of freshwater. At least nine families have small to large radiations in the freshwater environment: Corbiculidae, Sphaeriidae, Dreissenidae, and the unioniforme families: Hyriidae, Margaritiferidae, Unionidae, Etheriidae, Iridinidae, and Mycetopodidae. The unioniforme families contain at least 180 genera and about 800 species. This order is characterized by the unique parasitic larval stage on the gills, fins or the body of a particular host fish. This order of freshwater bivalves is suffering a very high rate of extinction, with about 37 species considered presumed extinct in North America alone. The level of endangerment and extinction facing these animals is primarily the result of habitat destruction or modification. Guest editors: E. V. Balian, C. Lévêque, H. Segers & K. Martens Freshwater Animal Diversity Assessment     

Genetic by environmental variation but no local adaptation in oysters (Crassostrea virginica)

Functional trait variation within and across populations can strongly influence population, community, and ecosystem processes, but the relative contributions of genetic vs. environmental factors to this variation are often not clear, potentially complicating conservation and restoration efforts. For example, local adaptation, a particular type of genetic by environmental (G*E) interaction in which the fitness of a population in its own habitat is greater than in other habitats, is often invoked in management practices, even in the absence of supporting evidence. Despite increasing attention to the potential for G*E interactions, few studies have tested multiple populations and environments simultaneously, limiting our understanding of the spatial consistency in patterns of adaptive genetic variation. In addition, few studies explicitly differentiate adaptation in response to predation from other biological and environmental factors. We conducted a reciprocal transplant experiment of first‐generation eastern oyster (Crassostrea virginica) juveniles from six populations across three field sites spanning 1000 km in the southeastern Atlantic Bight in both the presence and absence of predation to test for G*E variation in this economically valuable and ecologically important species. We documented significant G*E variation in survival and growth, yet there was no evidence for local adaptation. Condition varied across oyster cohorts: Offspring of northern populations had better condition than offspring from the center of our region. Oyster populations in the southeastern Atlantic Bight differ in juvenile survival, growth, and condition, yet offspring from local broodstock do not have higher survival or growth than those from farther away. In the absence of population‐specific performance information, oyster restoration and aquaculture may benefit from incorporating multiple populations into their practices.