Ultrastructural comparison of the spermatozoa of the Pacific oyster Crassostreagigas inhabiting polluted and relatively clean areas in Taiwan

The ultrastructure of spermatozoa of the Pacific oysters Crassostrea gigas from an industrially polluted area on the Hsinchu City coast and a relatively clean aquaculture area on Penghu Island, Taiwan, was studied. Oyster gonads were sectioned and examined with light and transmission electron microscopes. The number of spermatozoa in the acinus lumen was significantly lower in oysters from the polluted area than that from the relatively clean area (160 ± 33 cells per 0.01 mm² against 280 ± 42 cells per 0.01 mm², respectively, P < 0.01). Oysters from the polluted area on the Hsinchu City coast had 26.6% spermatozoa with damaged acrosome, coarsely granular chromatin and electron-lucent areas in the nucleus; cell membrane, mitochondria and flagellum were frequently absent in these spermatozoa. By contrast, oysters from the clean area on Penghu Island had 0.4% spermatozoa with the same impairments. Sea water temperature and salinity were similar in the two areas, whereas concentrations of nitrogenous nutrients and heavy metals (e.g., cadmium, copper, and zinc) were higher in the industrially polluted area. It is suggested that high percentage of impairments in the spermatozoa of oysters from the industrial area is attributable to environmental pollution.     

Use of encapsulated live microalgae to investigate pre-ingestive selection in the oyster Crassostrea gigas

The involvement of algal chemical cues in the pre-ingestive selection of food particles in Crassostrea gigas was studied using a new approach. Live cells of two microalgal species, Nitzschia closterium and Tetraselmis suesica, were separately entrapped in small alginate microcapsules using an emulsification/internal gelation method. Microcapsule size was adjusted to be within the range of particles ingested by oysters. Using this technique, about 80% of microcapsules had a diameter ranging from 21 to 100 μm. The monitoring of entrapped algae showed that phytoplankton cells remained alive and maintained an active growth for at least 24 days. In particle selection bioassays, adult C. gigas were fed a mixture of microcapsules containing the above algae species as well as control empty alginate microcapsules. The comparison of the proportions of each microcapsule type in the diet and in pseudofeces revealed that those containing T. suesica were significantly ingested while those containing N. closterium were preferentially rejected. Since microcapsule material (alginate matrix) prevented physical contacts between algae cells and oyster feeding organs, this study clearly demonstrate that extracellular metabolites produced by microalgae play a crucial role in the pre-ingestive selection of particles in suspension-feeding bivalves.     

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     

Influence of diet on pre-ingestive particle processing in bivalves: I: Transport velocities on the ctenidium

Suspension-feeding bivalve molluscs can assume a large ecological role by linking benthic and pelagic ecosystems. Therefore, a knowledge of the factors that influence feeding rates and processes at the level of the individual is important in understanding bivalve-dominated environments. We examined the roles of diet quality and concentration on particle processing by the ctenidia of four species of bivalves: the mussels Mytilus edulis L. and Mytilus trossulus Gould, and oysters Crassostrea virginica (Gmelin) and Crassostrea gigas (Thunberg). Bivalves were delivered diets of three different qualities at three different concentrations (1-2×10³, 10⁴, 10⁵ particles ml⁻¹). The high-quality diet consisted of the cryptophyte Rhodomonas lens Pascher et Ruttner; the low-quality diet consisted of particles prepared from ground Spartina sp. detritus; the medium-quality diet consisted of a 50:50 mixture (by particle number) of both particle types. Particle transport velocities on the ventral groove and dorsal tracts were then measured by means of video endoscopy. Ventral-groove transport velocities of M. edulis were the most responsive, demonstrating a significant increase with increasing diet quality, and a significant decrease with increasing diet concentration. Transport velocities in the ventral groove and dorsal tracts of C. virginica were not significantly affected by changes in diet quality, but significantly increased with increasing diet concentration. Transport velocities of M. trossulus and C. gigas demonstrated little change with diet quality or concentration, indicating that responses are species specific. Our data suggest that differential control of particle transport on the bivalve ctenidium is one of the underlying mechanisms that contribute to compensatory feeding responses exhibited by the entire organism.     

Genotoxicity biomarkers in aquatic bioindicators

Pollution of the aquatic environment is an ever-growing problem, as waters are the ultimate sink for the large number of xenobiotics from multiple sources. DNA damaging agents have a significant ecological relevance since they are implicated in many pathological processes and exert effects beyond that of individual being active through following generations. A large number of methods have been applied to evaluate genotoxic damage in different aquatic species. Comet assay, as method for de- tecting DNA alterations, and micronucleus test, as an index of chromosomal damage are the most widely applied and validated methods in field studies. These methods were applied in different vertebrate and invertebrate aquatic species, but only mollusk and fish species have been employed in routine biomonitoring programs. Mussels, due to their widely geographical distribution and the suitability for caging represent the bioindicator of choice in field studies. Mytilus species is the most used marine mussel. The use of fish is limited to specific geographic areas. The present review mainly focuses on the application of comet assay and micronucleus test in mussels. A number of biomonitoring studies in mussels, using comet assay or micronucleus test, revealed exposure to different classes of genotoxic compounds with a good discrimination power. The different evidence from the two as- says, reflects different biological mechanisms for the two genetic endpoints, DNA damage and chromosomal damage, suggesting their combined application in the field. Different endogenous and exogenous factors have been shown to modulate the genotoxic responses in mussels, acting as confounding factors in environmental monitoring. The use of standardized protocol for caging, sampling and genotoxity evaluation is critical in biomonitoring studies. The use of a multimarker approach coupling genotoxicity biomarkers with physiological and biochemical factors allows to have a complete picture of the environmental pollution [Current Zoology 60 （2）： 273-284, 2014].     

Isotopic signatures (δO and δC) of bivalve shells from cold seeps and hydrothermal vents

The oxygen and carbon isotopic compositions of 108 modern shells of various bivalve species collected from cold seeps and hydrothermal vents were investigated in order to evaluate whether these parameters can provide information on environmental geochemical variability as well as on bivalve species and on the type of symbiotic bacteria present in their gills. The results show that the carbonate of bivalve shells from hydrothermal vents is characterized by abnormal positive δ¹³C values due to kinetic isotope effects, whereas the carbonate of bivalve shells from cold seeps exhibits positive as well as negative δ¹³C values suggesting that oxidized methane emitted by the seeping fluids may be incorporated in the shell. Comparison of the δ¹⁸O and δ¹³C values of bivalve shells hosting different chemosymbiotic bacteria suggests that each type of symbiosis is associated with a specific environment and bivalve species, indicating that there is a strong physiological/metabolic control on the incorporation of stable isotopes during the biomineralization process.     

文蛤微卫星DNA的筛选及其特性分析

采用磁珠富集分离法从文蛤(Meretrix meretrix)的基因组中筛选得到49条微卫星DNA序列,其中两碱基重复有3种类型36个序列,四碱基重复有14种类型26个序列.重复次数在5～30之间的序列占75.6%,30次以上重复的序列占24.4%,最高重复次数为100.根据重复单元的排列特点,完美型、非完美型及混合型序列所占比例分别为61.2%、14.5%和24.5%.本研究中构建的文蛤微卫星文库将在文蛤种质资源评价及分子遗传学研究中发挥作用.     

Effects of GABA and epinephrine on the settlement and metamorphosis of the larvae of four species of bivalve molluscs

Competent larvae of different marine bivalve species were treated with GABA and epinephrine at different concentrations and times of exposure to test the ability of the drugs to induce settlement and metamorphosis. GABA induced both settlement and metamorphosis in the mussel Mytilus galloprovincialis, the clams Venerupis pullastra and Ruditapes philippinarum and the oyster Ostrea edulis. Maximum induction of settlement (>39%) was achieved after exposure of V. pullastra larvae to 10⁻⁴ M GABA; this concentration of GABA also induced the highest percentages of metamorphosis in the four species studied. Epinephrine was identified as an active inducer of settlement and metamorphosis in bivalve molluscs. Exposure to 10⁻⁵ M epinephrine induced significant levels of settlement in Mytilus, Venerupis and Ostrea. In contrast, epinephrine failed to induce settlement behaviour in Ruditapes. Maximum induction of metamorphosis was produced by 10⁻⁵ M epinephrine in mussels, clams and oysters; Ruditapes showed the highest percentage of metamorphosis (>78%). This is the first report in which the involvement of GABA in the settlement and metamorphosis of bivalve molluscan larvae is demonstrated. It was also recognised that epinephrine plays a role not only in inducing metamorphosis but also in initiating settlement.     

Linking taphonomy to community-level abundance: Insights into compositional fidelity of the Upper Triassic shell concentrations (Eastern Alps)

Although actualistic live/dead comparisons lead to robust estimates of fidelity of modern death assemblages, quantitative evaluation of fidelity of fossil assemblage remains uncertain. In this paper, effects of storm reworking on compositional fidelity of the Upper Triassic shell concentrations (Eastern Alps, Austria) are evaluated. An exploratory approach is based on comparison of reworked and non-reworked assemblages in ordination analyses. Non-reworked assemblages of one or more communities provide a baseline for evaluation of fidelity of reworked assemblages. In siliciclastic-rich intervals of the Kössen Formation, shell concentrations are represented by (1) packstones with small, shallow infaunal bivalves, (2) floatstones and pavements with large semi-infaunal bivalves, and (3) bioclastic marlstones. In carbonate-rich intervals, bioclastic floatstones with bivalves and brachiopods occur. Analyzing all shell concentrations, eight sample groups sharing similar species composition are discriminated. Limited effect of storm reworking on composition of shell concentrations is indicated by (1) a general persistence of six sample groups when only non-reworked assemblages are analyzed, (2) similarity in composition between reworked and non-reworked assemblages within sample groups, and (3) compositional segregation between non-reworked assemblages of distinctive sample groups, mostly without any reworked assemblages of intermediate composition.Depth-related variations in dead-shell production, shell destruction and body size governed preservation and distribution of the shell concentrations along onshore-offshore gradient in the Kössen Basin. First, at times when environmental conditions were unfavorable for shell producers, coupled with high background shell destruction rates, limestone beds formed during storm events were shell-poor. Second, less common shell concentrations in upper than in lower parts of siliciclastic intervals can be related to higher environmental stress in shallower habitats. Third, the difference between shell concentrations dominated by small and large bivalves is driven by between-habitat differences in body size and is not due to a differential sorting of small and large shells. Combining community analysis based on species abundances with taphonomic analysis can thus be helpful in tracking fidelity of fossil assemblages.     

Immunological and cytotoxicological responses of the Asian clam,Corbicula fluminea (M.), experimentally exposed to cadmium

Abstract

Bivalve molluscs, as filter-feeding organisms, are known to accumulate metals that can produce deleterious effects on organisms. The phagocytic activity of haemocytes and lysosomal alterations in the digestive gland cells were measured in the freshwater Asian clam exposed to cadmium, in order to assess the possible use of immunocompetence and lysosomal responses as biomarkers of freshwater quality. Clams were exposed in the laboratory to nominal concentrations of 3, 10, 21.4, 46.5 and 100 µg l^?1 of cadmium and sampled after 7, 15 and 30 days of exposure. The results show a decrease of phagocytic activity after only 7 days of exposure to 10 µg l^?1 of cadmium. This response was also observed as the exposure time was increased. Lysosomes in the digestive cells increased in size and number after 7 days of exposure as cadmium concentration increased. After 30 days of exposure, a decrease in size and number indicated a change in the response to the metal from concentrations of 46.5 µg l^?1 of cadmium. A dose and time response both in phagocytic activity of haemocytes and lysosomal structure demonstrated a possible use of these biomarkers in freshwater biomonitoring.