The contrasted evolutionary fates of deep‐sea chemosynthetic mussels (Bivalvia,Bathymodiolinae)

Bathymodiolinae are giant mussels that were discovered at hydrothermal vents and harboring chemosynthetic symbionts. Due to their close phylogenetic relationship with seep species and tiny mussels from organic substrates, it was hypothesized that they gradually evolved from shallow to deeper environments, and specialized in decaying organic remains, then in seeps, and finally colonized deep‐sea vents. Here, we present a multigene phylogeny that reveals that most of the genera are polyphyletic and/or paraphyletic. The robustness of the phylogeny allows us to revise the genus‐level classification. Organic remains are robustly supported as the ancestral habitat for Bathymodiolinae. However, rather than a single step toward colonization of vents and seeps, recurrent habitat shifts from organic substrates to vents and seeps occurred during evolution, and never the reverse. This new phylogenetic framework challenges the gradualist scenarios “from shallow to deep.” Mussels from organic remains tolerate a large range of ecological conditions and display a spectacular species diversity contrary to vent mussels, although such habitats are yet underexplored compared to vents and seeps. Overall, our data suggest that for deep‐sea mussels, the high specialization to vent habitats provides ecological success in this harsh habitat but also brings the lineage to a kind of evolutionary dead end.     

Parasitism in species of Bathymodiolus (Bivalvia: Mytilidae) mussels from deep-sea seep and hydrothermal vents

Bivalve species, especially mussels, are biomass dominants in many deep-sea chemosynthetic ecosystems. As in shallow-water environments, parasites are likely to be important factors in the population dynamics of bivalve communities in chemosynthetic ecosystems, but there has been little study of parasitism in deep-sea seep or vent molluscs. In this study, parasite types, diversity, prevalence, infection density and non-infectious indicators of stress or disease as related to host age, reproductive condition, and endosymbiont density were assessed in mussels (Bathymodiolus heckerae) from 2 seep sites and mussels (B. puteoserpentis) from 2 vent sites. We identified 10 microbial or parasitic agents in histological sections. Parasite types included 3 viral-like gut inclusions, 2 rickettsia-like gill inclusions, a rickettsia-like mantle inclusion, a bacterial gill-rosette, a chlamydia-like gut inclusion, gill-dwelling ciliates, and an unidentified inclusion in gut tissues. Parasite species richness was greater in seep mussels than in vent mussels, with the seep mussels possessing 9 types of parasites compared to 2 in the vent mussels. One of the viral-like inclusions infecting the seep mussel B. heckerae was pathogenic, causing lysis of the digestive tubules. The prevalence and intensity of infection by this pathogen were greater in hosts with shell lengths less than 100 mm. Mussels from all 4 sites also exhibited intense infiltration of tissues and blood spaces by enlarged hemocytes. Hemocytic infiltration (hemocytosis) showed variable degrees of severity that were not associated with other host factors examined.     

A hybrid zone between hydrothermal vent mussels (Bivalvia: Mytilidae) from the Mid-Atlantic Ridge

This study provides the first example of a hybrid zone between animal taxa distributed along the mid-ocean ridge system. We examined the distribution and genetic structure of deep-sea hydrothermal vent mussels (Bivalvia: Mytilidae) along a 2888-km portion of the Mid-Atlantic Ridge between 37 degrees 50' N and 14 degrees 45' N latitude. Mitochondrial DNA (mtDNA), allozymes and multivariate-morphometric evidence discriminated between individuals of a northern species, Bathymodiolus azoricus, and a southern species, B. puteoserpentis, that were separated by an intermediate ridge segment almost devoid of mussels. A small sample of mussels from Broken Spur, a vent locality along this intermediate zone, revealed a mixed population with gene frequencies and morphology that were broadly intermediate to those of the northern and southern species. Multilocus clines in mtDNA and allozyme frequencies were centred over the intermediate zone. We consider intrinsic and extrinsic processes that might limit genetic exchange across this hybrid zone.     

Genetics and taxonomy of Chilean smooth-shelled mussels, Mytilus spp. (Bivalvia: Mytilidae)

It has been previously established that native smooth-shelled mussels in southern South America possess close evolutionary affinities with Northern-Hemisphere Mytilus edulis L. 1758 (McDonald et al. (1991) [5]). This result has since been challenged by authors claiming that Chilean mussels should be considered a local subspecies of M. galloprovincialis Lmk. 1819. Moreover, morphological, physiological, ecotoxicological and molecular genetic studies on Chilean smooth-shelled mussels still frequently refer to ‘M. chilensis’ Hupé 1854, even though the previous discovery of alien M. galloprovincialis and considerable heterogeneity in shell morphology among samples collected along the Chilean shores raise concerns that different Mytilus spp. species might have been included under ‘M. chilensis’. Here we reviewed the molecular and morphological data available on smooth-shelled mussels from Chile in an attempt to clarify both their genetic composition and their taxonomic status. Using multivariate analysis on sample × allozyme-frequency matrices, we confirmed the widespread occurrence of the Southern-Hemisphere form of M. edulis along the shores from the North Patagonia region of Chile to the southern tip of the South American continent. The populations sampled in southern central Chile showed some evidence of slight introgression from Southern-Hemisphere M. galloprovincialis. Morphological characterization of a sample from Dichato in southern central Chile was consistent with its previous genetic identification as Mediterranean M. galloprovincialis. The occurrence of Southern-Hemisphere M. galloprovincialis in Punta Arenas at the southern tip of the South American continent was also reported. Southern-Hemisphere M. edulis, including native Chilean smooth-shelled Mytilus, should be assigned subspecific rank and named M. edulis platensis d’Orbigny 1846.     

Genetic diversity of Southern hemisphere blue mussels (Bivalvia: Mytilidae) and the identification of non‐indigenous taxa

The taxonomic and evolutionary affinities of Southern hemisphere smooth‐shelled blue mussels are unclear, with studies using different marker types having identified different relationships among various geographic regions. Using an existing and a new molecular assay, the present study builds on previous work to test the distribution of blue mussels native to and introduced to the Southern hemisphere. Populations of Mytilus were sampled from New Zealand, Australia, and Chile. The nuclear‐DNA marker Me 15/16 was used to identify the taxonomic status of 484 individuals. A new restriction fragment length polymorphism (RFLP) assay was used to identify the hemisphere of origin for a subset of Mytilus galloprovincialis. The Me15/16 marker identified 478 pure M. galloprovincialis from Southern hemisphere sites and six Mytilus edulis/M. galloprovincialis hybrids from the Auckland Islands (New Zealand) and Chile. A cytoplasmic RFLP identified Northern hemisphere M. galloprovincialis in almost every Southern hemisphere region. The presence of native M. galloprovincialis at high latitudes (up to 52°S) has implications for our understanding of environmentally induced selective constraints considered to determine species distributions. Widespread occurrence of invasive Northern hemisphere blue mussels in the Southern hemisphere is documented for the first time. Identification of inter‐specific hybrids (M. edulis × M. galloprovincialis) in Chile and in the Auckland Islands (subantarctic New Zealand) illustrates that environments ranging from international ports to remote protected locations are vulnerable to bioinvasion. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 898–909.     

Molecular taxonomy reveals broad trans-oceanic distributions and high species diversity of deep-sea clams (Bivalvia: Vesicomyidae: Pliocardiinae) in chemosynthetic environments

Large vesicomyid clams are common inhabitants of sulphidic deep-sea habitats such as hydrothermal vents, hydrocarbon seeps and whale-falls. Yet, the species- and genus-level taxonomy of these diverse clams has been unstable due to insufficiencies in sampling and absence of detailed taxonomic studies that would consistently compare molecular and morphological characters. To clarify uncertainties about species-level assignments, we examined DNA sequences from mitochondrial cytochrome-c-oxidase subunit I (COI) in conjunction with morphological characters. New and published COI sequences were used to create a molecular database for 44 unique evolutionary lineages corresponding to species. Overall, the congruence between molecular and morphological characters was good. Several discrepancies due to synonymous species designations were recognized, and acceptable species names were rectified with published COI sequences in cases where morphological specimens were available. We identified seven species with trans-Pacific distributions, and two species with Indo-Pacific distributions. Presently, 27 species have only been documented from one region, which might reflect limited ranges, or insufficient geographical sampling. Vesicomyids exhibit the greatest species diversity along the northwest Pacific ridge systems and in the eastern Pacific, along the western America margin, where depth zonation typically results in segregation of closely related species. The broad distributions of several vesicomyid species suggest that their required chemosynthetic habitats might be more common than previously recognized and occur along most continental margins.     

Detection of invasive and cryptic species in marine mussels (Bivalvia,Mytilidae): A chromosomal perspective

Marine mussels illustrate a stunning variability in shape and color. Such variability, added to the scarcity of reliable morphological characters for their identification, can mislead recognition prompting the assignation of specimens of a single species to different ones or incorporate specimens belonging to different taxa into a single one. DNA barcoding is widely used for species identification; however, as this method relies on the previous morphological identification of the specimens, some of the DNA sequences stored in DNA databases are incorrectly assigned to a given species. In view of this uncertainty, further criteria beyond morphological characters and DNA sequences in databases are required to more reliably and accurately identify marine mussels. In this work we mapped ribosomal RNA and histone gene clusters to chromosomes of four species of marine mussels and compared them with those from another eight marine mussel taxa. Specimens of these twelve taxa were also DNA barcoded. Our results clearly demonstrated that the chromosomal analysis of marine mussels could shed light on their identification and, therefore, solve contradictions posed by morphological and molecular data.     

Toxins-antitoxins: diversity, evolution and function

Genes for toxin-antitoxin (TA) complexes are widespread in prokaryote genomes, and species frequently possess tens of plasmid and chromosomal TA loci. The complexes are categorized into three types based on genetic organization and mode of action. The toxins universally are proteins directed against specific intracellular targets, whereas the antitoxins are either proteins or small RNAs that neutralize the toxin or inhibit toxin synthesis. Within the three types of complex, there has been extensive evolutionary shuffling of toxin and antitoxin genes leading to considerable diversity in TA combinations. The intracellular targets of the protein toxins similarly are varied. Numerous toxins, many of which are sequence-specific endoribonucleases, dampen protein synthesis levels in response to a range of stress and nutritional stimuli. Key resources are conserved as a result ensuring the survival of individual cells and therefore the bacterial population. The toxin effects generally are transient and reversible permitting a set of dynamic, tunable responses that reflect environmental conditions. Moreover, by harboring multiple toxins that intercede in protein synthesis in response to different physiological cues, bacteria potentially sense an assortment of metabolic perturbations that are channeled through different TA complexes. Other toxins interfere with the action of topoisomersases, cell wall assembly, or cytoskeletal structures. TAs also play important roles in bacterial persistence, biofilm formation and multidrug tolerance, and have considerable potential both as new components of the genetic toolbox and as targets for novel antibacterial drugs.     

Toxins-antitoxins: diversity,evolution and function

Genes for toxin-antitoxin (TA) complexes are widespread in prokaryote genomes, and species frequently possess tens of plasmid and chromosomal TA loci. The complexes are categorized into three types based on genetic organization and mode of action. The toxins universally are proteins directed against specific intracellular targets, whereas the antitoxins are either proteins or small RNAs that neutralize the toxin or inhibit toxin synthesis. Within the three types of complex, there has been extensive evolutionary shuffling of toxin and antitoxin genes leading to considerable diversity in TA combinations. The intracellular targets of the protein toxins similarly are varied. Numerous toxins, many of which are sequence-specific endoribonucleases, dampen protein synthesis levels in response to a range of stress and nutritional stimuli. Key resources are conserved as a result ensuring the survival of individual cells and therefore the bacterial population. The toxin effects generally are transient and reversible permitting a set of dynamic, tunable responses that reflect environmental conditions. Moreover, by harboring multiple toxins that intercede in protein synthesis in response to different physiological cues, bacteria potentially sense an assortment of metabolic perturbations that are channeled through different TA complexes. Other toxins interfere with the action of topoisomersases, cell wall assembly, or cytoskeletal structures. TAs also play important roles in bacterial persistence, biofilm formation and multidrug tolerance, and have considerable potential both as new components of the genetic toolbox and as targets for novel antibacterial drugs.     

Genetic and morphometric characterization of mussels (Bivalvia: Mytilidae) from mid-atlantic hydrothermal vents.

Mussels were collected from deep-sea hydrothermal vents along the Mid-Atlantic Ridge. Specimens from the Snake Pit site were previously identified genetically and anatomically as Bathymodiolus puteoserpentis, but the relationships of mussels from other sites (Logatchev and Lucky Strike) were unclear. Molecular genetic and morphological techniques were used to assess differences among these mussel populations. The results indicate that the range for B. puteoserpentis extends from Snake Pit to Logatchev, and that an unnamed second species, B. n. sp., occurs at Lucky Strike. Analysis of mitochondrial NADH dehydrogenase subunit 4 (ND4) revealed 13% sequence divergence between the two species. Nei's genetic distance (D) based on 14 allozyme loci was 0.112. A multivariate morphometric analysis yielded a canonical discriminant function that correctly identified individuals from these sites to species 95% of the time.     

Influence of salinity on the physiological conditions in mussels, Perna perna and Perna viridis (Bivalvia: Mytilidae)

Perna genus was introduced to Venezuela, but nowadays, Perna perna and Perna viridis coexist and are commercially exploited from their natural beds. The aim of this work was to determine the effect of salinity on the physiological conditions of these species by studying RNA/DNA and Protein/DNA ratios. The organisms were collected from natural beds at La Esmeralda, Sucre State, Venezuela, and acclimatized for 15 days under laboratory conditions at 25 degrees C, 36 per thousand salinity, pH between 7 and 8 and more than 90% of oxygen saturation. Later, they were divided in two groups: for one group, the salinity concentration was increased (36 to 45 per thousand), and for the other, the salinity was decreased (36 to 15 per thousand). The rate of change was 1 per thousand every day. Ten organisms per group of both species were taken at each of 15, 20, 25, 30, 36, 40 y 45 per thousand salinity concentrations. Protein (colorimetric method) and nucleic acids (RNA and DNA by fluorometric method) concentrations were measured in the digestive gland, gills and adductor muscle tissues. Results indicate that P. perna can physiologically compensate the increase in salinity, but not when the salinity decreased, when proteins are the most needed macromolecules. The Protein/DNA index is directly related to salinity changes in both cases. P. viridis shows physiological compensation to salinity increases and decreases. The RNA/DNA index value in both cases supports this. Digestive gland and muscle tissues are the regulating tissues in both species. These results show that P. viridis has a higher degree of adaptability to salinity changes and, therefore, a greater potential for aquaculture than P. perna.     

On the evolutionary ecology of symbioses between chemosynthetic bacteria and bivalves

Mutualistic associations between bacteria and eukaryotes occur ubiquitously in nature, forming the basis for key ecological and evolutionary innovations. Some of the most prominent examples of these symbioses are chemosynthetic bacteria and marine invertebrates living in the absence of sunlight at deep-sea hydrothermal vents and in sediments rich in reduced sulfur compounds. Here, chemosynthetic bacteria living in close association with their hosts convert CO₂ or CH₄ into organic compounds and provide the host with necessary nutrients. The dominant macrofauna of hydrothermal vent and cold seep ecosystems all depend on the metabolic activity of chemosynthetic bacteria, which accounts for almost all primary production in these complex ecosystems. Many of these enigmatic mutualistic associations are found within the molluscan class Bivalvia. Currently, chemosynthetic symbioses have been reported from five distinct bivalve families (Lucinidae, Mytilidae, Solemyidae, Thyasiridae, and Vesicomyidae). This brief review aims to provide an overview of the diverse physiological and genetic adaptations of symbiotic chemosynthetic bacteria and their bivalve hosts.     

Linking fisheries management and conservation in bioengineering species: the case of South American mussels (Mytilidae)

We examined a complete list of South American mussels (Mytilidae) to identify species with current or potential needs for management and conservation actions. Based on ecological/ecosystem (aggregations, beds or banks affecting ecosystem functioning) and socio-economic (artisanal fisheries or aquaculture systems) attributes species with high relevance were identified. At least 14 species exhibited large ecosystem level effects at local scales. Further, most of them also sustain important fisheries: nearly one/third of these fisheries showed characteristics that may contribute to their lack of sustainability and overexploitation, while half are either in the initial exploitation phase or in the stabilization of extraction and institutionalization phase. Invading species are modifying the structure of mussel habitats. Allocation of spatially explicit management tools, notably Territorial User Rights in Fisheries and Marine Reserves, together with co-management initiatives, are suggested as relevant tools to fulfill management and conservation objectives for these key bioengineering species.     

Dispersal barriers and isolation among deep-sea mussel populations (Mytilidae: Bathymodiolus) from eastern Pacific hydrothermal vents

Deep-sea hydrothermal vent species are widely dispersed among habitat islands found along the global mid-ocean ridge system. We examine factors that affect population structure, gene flow and isolation in vent-endemic mussels of the genus Bathymodiolus from the eastern Pacific Ocean. Mussels were sampled from localities including the Galapagos Rift (GAR, 0 degrees 48' N; 86 degrees 10' W) and the East Pacific Rise (EPR, 13 degrees N to 32 degrees S latitude) across a maximum distance of 4900 km. The sampled range crossed a series of topographical features that interrupt linear aspects of the ridge system, and it encompassed regions of strong cross-axis currents that could impede along-axis dispersal of mussel larvae. Examinations of mitochondrial DNA sequences and allozyme variation revealed significant barriers to gene flow along the ridge axis. All populations from the GAR and EPR from 13 degrees N to 11 degrees S were homogeneous genetically and appeared to experience unimpeded high levels of interpopulational gene flow. In contrast, mussels from north and south of the Easter Microplate were highly divergent (4.4%), possibly comprising sister-species that diverged after formation of the microplate approximately 4.5 Ma. Strong cross-axis currents associated with inflated bathymetry of the microplate region may reinforce isolation across this region.     

Tissue-specific expression of male-transmitted mitochondrial DNA and its implications for rates of molecular evolution in Mytilus mussels (Bivalvia: Mytilidae).

Mytilus and other bivalves exhibit an unusual system of mitochondrial DNA (mtDNA) transmission termed doubly uniparental inheritance (DUI). Specifically, males transmit the mtDNA they have received from their fathers to their sons. Females transmit their mother's mtDNA to both sons and daughters. Males are normally heteroplasmic and females are normally homoplasmic, but not exclusively. This system is associated with an unusual pattern of molecular evolution. Male-transmitted mtDNA (M type) evolves faster than female-transmitted (F type) mtDNA. Relatively relaxed selection on the M type has been proposed as an explanation for this phenomenon. To further evaluate the selective forces acting upon the M-type genome, we used RT-PCR to determine where it is expressed. M-type mtDNA expression was detected in all gonad samples and in 50% of somatic tissues of males, and in a single female tissue. F-type mtDNA expression was detected in all female tissues, all male somatic tissues, and all but one male gonad sample. We argue that the expression of M-type mtDNA in male somatic and male gonad tissues has implications for the strength of selection acting upon it.     

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     

Phylogenetic characterization of three morphs of mussels (Bivalvia, Mytilidae) inhabiting isolated marine environments in Palau Islands

Marine lakes in the Palau Islands are known to harbor unique marine fauna that have remained isolated since the formation of the lakes after the Last Glacial Maximum. We analyzed mussels from marine lakes located on different islands and conducted morphological, phylogenetic and population genetic characterization to clarify their evolutionary history. The mussels were morphologically classified into three differentiated morphs: NS, ON, and MC. Their common characteristics were consistent with the Brachidontes-Hormomya complex of the Mytilidae family. Phylogenetic analysis based on the nuclear 18S ribosomal RNA gene supported the taxonomic position of the mussels among the Mytilidae. In the mitochondrial cytochrome c oxidase subunit I (COI) gene lineage, NS-and MC-morphs were highly diverged from each other; their estimated time of divergence dates back to the mid-Pleistocene. ON-morph was more closely related to MC-morph, although the shell morphologies of ON- and MC-morphs were easily distinguishable. Population genetic analysis revealed the coexistence of highly diverged haplotypes within a population of ON-morph, indicating introgression of mtDNA among the morphs. Our data suggest that morphological differentiation of marine lake mussels can occur in a relatively short period under different environmental conditions. Thus, the marine lakes provide a unique site for the study of diversification in mussels.     

印度洋深海沉积物石油烃降解菌分离、鉴定与多样性分析

烃类物质在海洋环境中广泛分布,深海可能含有特殊的烃降解微生物。本研究通过对西南印度洋中脊与印度洋中部深海沉积物中石油降解菌的富集培养和分离鉴定,从6个站点的样品中共分离得到800株菌,通过BOX-PCR去重复菌株后,对其中183株菌进行了16S rRNA基因序列分析,发现这些菌分属于23个属;其中,γ-变形菌纲的食烷菌属(Alcanivorax)和放线菌纲的微杆菌属(Microbacterium)占优势。此外,还发现了食烷菌属2个潜在的新种、假海栖菌属(Pseudooceanicola)1个潜在新种。高通量测序结果证明,富集菌群中γ-变形菌纲是优势菌,主要包括食烷菌属、盐单胞菌属(Halomonas)、海杆菌属(Marinobacter)等。结合可培养菌与高通量测序结果,食烷菌属、盐单胞菌属、海杆菌属、交替假单胞菌(Pseudoalteromonas)、海源菌属(Idiomarina)与微杆菌属(Microbacterium)是沉积物样品中常见的石油烃降解菌,迪茨氏菌属(Dietzia)、红球菌属(Rhodococcus),假单胞菌属(Pseudomonas)、赤杆菌属(Erythrobacter)与副球菌属(Paracoccus)等可能也参与了烃的降解。     

早期形态发生:一种解决贻贝科(软体动物门:双壳纲)分类、系统发育和进化问题的方法

作者对贻贝科贝类的幼虫和幼贝期发育阶段形态结构的出现和变化顺序进行了研究,其约60个不同分类单元的个体发生可归纳为4种形态发生类型或模式。主要对3个形态发生区域的阶段形态结构的起源、发育变化和同源性做了研究。其一,即中央区域,开始形成于前双壳Ⅰ期(PD-Ⅰ),在某个分类单元它可以在前双壳Ⅱ期(PD-Ⅱ)和幼贝期(N)形成,而在其它分类单元则在前双壳Ⅱ期、幼贝期和双壳期(D)形成;第二区域,即背部后区,在幼贝期出现;第三区域,即背部前区,出现于双壳期。双壳期背部后区在某个分类单元起源于幼贝期的形态构造,在其它分类单元则可能起源于双壳期的形态构造。与在贻贝分类学上应用的成体特征相比,早期发育阶段中央和背部后区的形态结构显示出很明显的发育顺序或特征变化规律。根据以前人们熟知而尚未应用到分类和系统发育研究中的早期发育阶段形态特征,作者重新修订了Soot-Ryen的现生贻贝科种上阶元分类系统,重新提出了科内系统发育关系。修订的分类系统表明,Scarlato and Starobogatov(1984)提出的贻贝科各亚科由偏顶蛤亚科开始,沿4条系统发育路线演化发展,对应其早期发育阶段的4类形态发生类型或模式。