Evolution of active host-attraction strategies in the freshwater mussel tribe Lampsilini (Bivalvia: Unionidae)

Most freshwater mussels (Bivalvia: Unionoida) require a host, usually a fish, to complete their life cycle. Most species of mussels show adaptations that increase the chances of glochidia larvae contacting a host. We investigated the evolutionary relationships of the freshwater mussel tribe Lampsilini including 49 of the approximately 100 extant species including 21 of the 24 recognized genera. Mitochondrial DNA sequence data (COI, 16S, and ND1) were used to create a molecular phylogeny for these species. Parsimony and Bayesian likelihood topologies revealed that the use of an active lure arose early in the evolution of the Lampsiline mussels. The mantle flap lure appears to have been the first to evolve with other lure types being derived from this condition. Apparently, lures were lost independently in several clades. Hypotheses are discussed as to how some of these lure strategies may have evolved in response to host fish prey preferences.     

Resolving species boundaries in the critically imperiled freshwater mussel species,Fusconaia mitchelli (Bivalvia: Unionidae)

Species are a fundamental unit of biology, and defining accurate species boundaries is integral to effective conservation and management of imperiled taxa. Freshwater mussels (Bivalvia: Unionidae) are among the most imperiled groups of organisms in North America, yet species boundaries remain uncertain for many taxa. The False Spike, Fusconaia mitchelli (Simpson in Dall, 1895), is a freshwater mussel considered to be endemic to central Texas (Brazos, Colorado, and Guadalupe drainages). Recent research revealed significant intraspecific genetic variation between geographically separated populations of F. mitchelli, which could be indicative of speciation; however, small sample sizes for several of the populations precluded formal taxonomic revision. Here, we increase taxon sampling and use multilocus DNA sequence data and traditional morphometrics to re‐evaluate species boundaries in F. mitchelli. We sequenced three loci: the protein‐coding mitochondrial DNA genes cytochrome c oxidase subunit 1 and NADH dehydrogenase 1, and the nuclear internal transcribed spacer 1. Phylogenetic analyses depicted deep genetic divergence between F. mitchelli in the Guadalupe and those in the Brazos and Colorado drainages, which was further supported by available biogeographic information. Morphometric analyses and coalescent‐based species delimitation models integrating both DNA sequence and morphological data provided strong support for the divergence observed between the two geographically isolated clades of F. mitchelli. Based on these results, we revise taxonomy accordingly by elevating the junior synonym Fusconaia iheringi (Wright, 1898) to represent the Brazos and Colorado populations and restrict the distribution of F. mitchelli to the Guadalupe River drainage. Our findings may impact pending management decisions to protect F. mitchelli under the U.S. Endangered Species Act.     

Climate effects on inter‐annual variation in growth of the freshwater mussel (Anodonta beringiana) in an Alaskan lake

1. Warming trends are evident in many parts of the globe but are especially marked at higher latitudes, with complex effects on the biota that include direct effects on growth potential and indirect effects through food webs. 2. Air temperatures have been increasing over the past 50 years in southwestern Alaska, affecting the growth and population dynamics of many organisms, including a variety of aquatic species such as the freshwater mussel Anodonta beringiana. 3. We collected freshwater mussels from Iliamna Lake, in the Bristol Bay region of Alaska, and measured their shells to examine climatic effects on growth patterns. 4. Linear mixed effects models and ordinary least square linear regressions revealed strong positive correlations between local air temperatures (especially in May, October and the summer months) and inter‐annual variation in mussel growth. Slower mussel growth was also significantly correlated with later date of ice break‐up, which was linked to air temperatures in late spring.     

Sediment particles as a cause of nacre staining in the freshwater mussel,Amblema plicata (Say) (Bivalvia: Unionidae)

Brown stains sometimes appear in the inner shell layers (nacre) of freshwater mussels. An electron microprobe was used to analyze the stained nacre of the unionid Amblema plicata (Say, 1817) from selected localities on the Mississippi River in the vicinity of LaCrosse and Prairie du Chien, Wisconsin.Several elements such as Na, Mg, Al, Si, P, S, Cl, K, and Fe are more highly concentrated in stained than in unstained nacre. Concentrations of these elements relative to Ca were found to vary significantly among the localities from which the specimens were obtained. Ratios have significantly higher variances downstream of the confluence of the Yellow and Mississippi Rivers, downstream of a barge fleeting area, near the town of Marquette, Iowa, near the site of sewage effluent for Prairie du Chien, Wisconsin and downstream of a scrap metal yard near LaCrosse, Wisconsin in contrast to control localities.Suspended silt, the result of runoff and river activity (barge traffic, dredging, pleasure boating) may be the stimulus for stain formation. Clay minerals adsorb accessory elements and in turbid water are trapped within the pallial space of A. plicata. The mussel secretes an organic-rich, periostracum-like layer over the entrapped sediment, and later reverts to prismatic and finally nacreous shell deposition. Some of the elements found in the stain could directly disturb Ca metabolism by competing with Ca for binding sites in shell aragonite.     

Growth patterns of the pan‐European freshwater mussel,Anodonta anatina (Linnaeus, 1758) (Bivalvia: Unionidae), vary with sex and mortality in populations

1. Post‐maturation growth leading to indeterminate growth patterns is widespread in nature. However, its adaptive value is unclear. Life history theory suggests this allocation strategy may be favored by temporal pulses in the intensity of mortality and/or the capacity to produce new tissues.
2. Addressing the origin of indeterminate growth and the variability of growth patterns, we studied the growth of duck mussels, Anodonta anatina, a pan‐European unionid, in 18 Polish lakes. For each population, the sex, size, and age of collected mussels were measured to estimate Bertalanffy''s growth curve parameters. We integrated information on A. anatina mortality rates, lake trophy, biofouling by zebra mussels, Dreissena polymorpha, and the prevalence of parasitic trematode larvae to identify selective conditions in lakes.
3. We found two sources of mortality in A. anatina populations, pertaining to adverse effects of zebra mussel biofouling and trophy state on mussel survival. Additionally, populations with heavier biofouling presented a smaller abundance of parasites, indicative of a relationship between filtering intensity and contraction of water‐borne trematode larvae by filtering A. anatina.
4. Consistently for each sex, populations with a greater trophy‐related mortality were characterized in A. anatina by a smaller asymptotic size L_max, indicative of a life history response to mortality risk involving early maturation at a smaller body size. In all populations, females featured higher mortality and larger asymptotic size versus males.
5. Our findings support a theoretical view that adaptive responses to selection involve adjustments in the lifetime resource allocation patterns. These adjustments should be considered drivers of the origin of indeterminate growth strategy in species taking parental care by offspring brooding in body cavities.

     

Isolation and characterization of fifteen polymorphic microsatellite loci in the threatened freshwater mussel Solenaia oleivora (Bivalvia: Unionidae)

Freshwater mussels are among the most endangered fauna in the world, while little is known about their genetic diversity. We isolated 15 polymorphic microsatellite loci in a threatened freshwater mussel, Solenaia oleivora. The characteristics of these loci were tested with 46 individuals. The number of alleles per locus ranged from 2 to 15, and the PIC value ranged from 0.250 to 0.880. The observed and expected heterozygosity ranged from 0.167 to 0.929 and from 0.297 to 0.900, with an average of 0.562 and 0.647, respectively. Eight loci deviated significantly from Hardy–Weinberg equilibrium. These polymorphic microsatellites will be useful for the assessment of population genetic diversity and resource conservation of S. oleivora. Cross-species amplification demonstrated that six of 15 microsatellite loci amplified successfully in two other unionid species.     

Testing hypotheses on the dispersal and evolutionary history of freshwater mussels (Mollusca: Bivalvia: Unionidae)

The relationship between dispersal and differentiation of the European freshwater mussel Unio pictorum (Linnaeus, 1758) was studied with molecular genetic methods. Forty‐two populations from France, Italy and central Europe were analysed. Genetic relationships were assessed from the geographical distribution of allele frequencies at 17 enzyme loci. Neighbouring groups of populations show small to moderate mean genetic distances (0.020 < D_mean < 0.263). With a few exceptions the genetic affinities of the populations are the closest within the same drainage basin. In central Europe and Northern Italy genetic differences between drainage systems are relatively large. Populations from north‐eastern Italy are genetically similar to Danubian populations. Mussels from the islands of Corsica and Sardinia are more closely related to populations from the Italian peninsula than to French populations from the Rhône drainage system. Genetic relationships within U. pictorum from central Europe reflect palaeogeographical relationships between river systems during the Pliocene and Pleistocene. Literature data on two North American unionid species and one European fish species show the same relationship between genetic diversity and the history of drainage systems, although the correlations are less strong. In France and Italy this correspondence is much less evident. Population dynamic processes and human activities leading to populational bottlenecks might have obscured it.     

Genetic variability among freshwater mussel Anodonta woodiana (Lea, 1834) (Bivalvia: Unionidae) populations recently introduced in Poland

The alien Chinese mussel Anodonta woodiana was first reported in Poland in the system of heated lakes near Konin in 1993. Genetic studies with use of three molecular techniques (isoenzyme electrophoresis, PCR-RFLP and sequence analysis of a COI gene fragment) were carried out on the Polish first populations of A. woodiana. The studies have revealed low genetic variation between the populations (Nei's genetic distance for 12 loci ranged 0.000 to 0.007) as well as their considerable polymorphism. Each population averaged 2.28 alleles per locus, 2.72 alleles per polymorphic locus, and 75% polymorphic loci. Restriction analysis of the COI gene fragment have not revealed variability between the analysed specimens, including males and females. Restriction enzymes, ScrFI, Csp6I, and EcoRI used in the COI gene fragment PCR-RFLP generate distinct restriction patterns, which can be molecular markers for A. woodiana. The sequence obtained for COI fragment was the same in the examined female and male specimens and represents F mitotype (DNA was isolated from somatic tissues). The divergence between A. woodiana F and M mitotypes is high (34%), however it remains within the range of the general character of the DUI (doubly uniparental inheritance) phenomenon in freshwater bivalves (Unionidae).     

An integrative approach underscores the taxonomic status of Lamellidens exolescens,a freshwater mussel from the Oriental tropics (Bivalvia: Unionidae)

The Oriental Region harbours the second richest fauna of freshwater bivalves in the world, including many endangered endemic taxa. However, the Oriental fauna of the Unionidae have been very poorly studied using an integrative taxonomic approach, which may provide reasonable revisions of complicated (cryptic) taxa based on morphological, molecular, biogeographic and ecological evidence. Here, we present the first example of an integrative taxonomic revision concerning the status of Unio exolescens Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]), a nominal mussel taxon that was accepted as a valid species within the genus Trapezoideus Simpson (1900). Currently, Trapezoideus exolescens is considered the type of the genus as far as the originally designated type species, U. foliaceus Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]), was considered to be a synonym of T. exolescens. Using nucleotide sequences obtained from mitochondrial (COI and 16S rRNA) and nuclear (28S rDNA) genes, we found that the topotypes of Unio exolescens Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) cluster together with representatives of another mussel genus, Lamellidens Simpson (1900). Based on these results and on morphological data, we transfer Unio exolescens Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) from Trapezoideus to Lamellidens and propose Lamellidens exolescens (Gould, 1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) comb. nov. In addition, we revisited the status of Unio foliaceus Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) as a valid species and the type of the genus Trapezoideus based on the morphological study of the type specimen, although a question concerning the true position of this taxon is still open because its molecular sequences are not available. Our findings highlight that an integrative taxonomic approach is an important tool, particularly when dealing with such species-rich Unionidae fauna as those of the Oriental Realm.     

Byssogenesis in the juvenile pink heelsplitter mussel,Potamilus alatus (Bivalvia: Unionidae)

The North American pink heelsplitter (Potamilus alatus) differs from most freshwater mussels in China by the ability to secrete an ephemeral byssus during its juvenile stage. In the present study, light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were used to investigate this ephemeral byssal structure, and amino acid composition was analyzed and compared with that of other species. The results revealed that the byssus consists of a long byssal thread and a few adhesive plaques which are randomly set up along the thread and assembled by petioles. There is a thin but distinctive cuticle with a continuous homogeneous matrix surrounding the byssal thread. Structural variation occurred when the byssal thread was differentially stretched. Four‐stranded helical primary fasciculi, which form a stable rope‐like structure, become evident after removal of the cuticle. The primary fasciculi consist of bundles of hundreds of parallel secondary fasciculi, each measuring about 5 μm in diameter. All evidence indicates that the byssus of the pink heelsplitter has a significantly different macrostructure and microstructure than the permanent byssus of the marine mussel Mytilus. Byssogenesis ceases when juveniles exceed 30 mm in length, although it varies greatly even among juveniles of similar size. Byssus formation is influenced by substrate type. The unique characteristics of the byssus have important advantages for survival, transition, and aggregation during the early life history. This study not only provides first insight into the structure of the ephemeral byssus and its relationship to freshwater mussel development and growth, but also suggests possibilities for the synthesis of novel biopolymer materials particularly useful in freshwater ecosystems. J. Morphol. 276:1273–1282, 2015. © 2015 Wiley Periodicals, Inc.     

Revisiting the North American freshwater mussel genus Quadrula sensu lato (Bivalvia Unionidae): Phylogeny,taxonomy and species delineation

Freshwater mussels (Bivalvia, Unionidae) have suffered strong declines over the last century. High morphological plasticity of Unionidae causes disturbances in their systematics and taxonomy, hampering conservation efforts. Species that have historically been placed under the North American genus Quadrula have suffered from numerous taxonomic and species delineation problems since its inception. Four genera are presently recognized within Quadrula sensu lato, that is, Cyclonaias, Quadrula, Theliderma and Tritogonia, but their phylogenetic basis remains incompletely tested. In the present study, we reconstructed several two‐marker (mtDNA cytochrome c oxidase subunit I—COI and NADH dehydrogenase subunit 1—ND1) phylogenies with newly collected specimens and all previously available sequences covering most species within this group. We then delineated the species within the group using an integrative approach with the application of molecular statistical methods, morphometric (Fourier Shape) analyses and geographic distribution data. Four clades corresponding to these genera were consistently recovered in all phylogenies. To validate the generic status of these clades, molecular analyses were complemented with morphological, anatomical and ecological data compiled from the literature. Several revisions are here proposed to the current systematics and taxonomy of these genera, including the synonymization of Cyclonaias asperata under Cyclonaias kieneriana; the inclusion of Quadrula apiculata and Quadrula rumphiana under Quadrula quadrula; the placement of Quadrula nobilis under Tritogonia; and finally the separation of the Mobile River basin populations of Theliderma metanevra as a new species, that is, Theliderma johnsoni n. sp. The conservation implications of the proposed changes are then discussed.     

Estimating the biomass of freshwater mussels (Bivalvia: Unionidae) from shell dimensions

Packs of autumn-shed maple leaves were placed at coal ash effluent-exposed and reference sites in streams on December 5, 1977 and removed after 27 and 96 days. Leaf surface area (cm²/leaf) and disc weight (ash-free dry wt/15 mm disc) were greater at the effluent-exposed site than at the reference site after 96 days (p < .001). ATP content of leaves from the reference stream quadrupled between 27 and 96 days while ATP content of effluent-exposed leaves remained low. Macroinvertebrates colonized the leaf packs in the reference site but were not found on or in effluent-exposed packs. We concluded that leaf processing beyond the leaching of soluble organics did not occur in the effluent-exposed packs owing to reduced colonization and decomposition by fungi. Since stream invertebrates prefer decomposed leaf material and animals grow faster on leaves colonized by microbes, the ash effuent appears to indirectly affect macroinvertebrates by interfering with leaf decomposition and thus reducing the quality of their food.     

Integrative taxonomy reveals phenotypic plasticity in the freshwater mussel Contradens contradens (Bivalvia: Unionidae) in Thailand,with a description of a new species

The freshwater mussel Contradens contradens (Lea, 1838) occurs in most types of freshwater habitats throughout Thailand. The species shows extensive variation in shell morphology, which has led to the recognition of six different subspecies. In this study, the validity of these six subspecies plus one unknown species was assessed using an integrative taxonomic approach. Geometric morphometric analyses revealed significant differences in shell shapes among these six nominal morphological subspecies, although a considerable degree of overlap was detected in some groups. In contrast, the phylogenetic tree obtained from the concatenated data of mitochondrial COI and nuclear H3 gene sequences and molecular species delimitation analyses revealed only three supported clades. These clades are proposed herein as three distinct species, and strongly corresponded to the biogeographically disjunct drainage systems in Thailand. They consisted of the (i) C. contradens clade found in the Chao Phraya Basin and other rivers that drain into the Gulf of Thailand, (ii) C. crossei clade that is restricted to the Middle Mekong Basin, and (iii) a clade containing only the newly discovered species from Huai Luang River in the north-east of Thailand, which is described herein as Contradens rolfbrandti Jeratthitikul & Panha, sp. nov. Speciation among these congeners was probably caused by the restriction of gene flow due to the past geomorphology of the river systems. The intraspecific variation in the shell shape detected here does not reflect the evolution of the mussel, but rather is evidence of phenotypic plasticity.http://zoobank.org/urn:lsid:zoobank.org:pub:649B6093-E1DD-4FD8-8185-A4696C43AD36; http://zoobank.org/urn:lsid:zoobank.org:act:777DDE86-8397-4FF3-AA78-0BE0F34611F4     

Evolution of bilaterally asymmetrical larvae in freshwater mussels (Bivalvia: Unionoida: Unionidae)

Bilaterally asymmetrical glochidia (i.e. bivalved parasitic larvae bearing a large marginal appendage on a single valve) have been reported from five Asian freshwater mussel genera belonging to two separate subfamilies, the Gonideinae (i.e. Pseudodon, Solenaia, and Physunio) and Rectidentinae (i.e. Contradens and Trapezoideus). This classification requires that the bilaterally asymmetrical glochidium‐bearing mussels are not monophyletic, and suggests that this atypical larval morphology evolved twice in the same geographic region. Although homoplastic glochidium characters are known (e.g. marginal appendages and size), we hypothesized that bilaterally asymmetrical glochidia represent a novel morphological synapomorphy. We tested the monophyly of the mussels bearing bilaterally asymmetrical glochidia using a molecular matrix consisting of representatives from all six freshwater mussel families and three molecular markers (28S, 16S, and COI). Bayesian inference, maximum likelihood, and ancestral state reconstruction were employed to estimate the phylogeny and larval trait transformations. The reconstructed phylogeny rejects the monophyly of the asymmetrical glochidium‐bearing mussels and resolves two putative origins of asymmetrical glochidia; however, ancestral state reconstruction supports asymmetrical glochidia as a synapomorphy of only one supraspecific taxon of the Rectidentinae. In the Gonideinae, asymmetrical glochidia were autapomorphic of Pseudodon cambodjensis (Petit, 1865). That is, no other taxa resolved among the Gonideinae had bilaterally asymmetrical glochidia, including other Pseudodon species. We describe how the alleged intrageneric glochidial variation in Pseudodon, and in the other genera of the Gonideinae reported to have asymmetrical glochidia (i.e. Solenaia and Physunio), challenge the resolved convergence of asymmetrical glochidia. Our results are discussed in the context of freshwater mussel larval evolution, patterns in life‐history traits, and the classification of freshwater mussels generally. © 2015 The Linnean Society of London     

Incongruence between mtDNA and nuclear data in the freshwater mussel genus Cyprogenia (Bivalvia: Unionidae) and its impact on species delineation

Accurately identifying species is a crucial step for developing conservation strategies for freshwater mussels, one of the most imperiled faunas in North America. This study uses genetic data to re‐examine species delineation in the genus Cyprogenia. Historically, Cyprogenia found west of the Mississippi River have been ascribed to Cyprogenia aberti (Conrad 1850 ), and those east of the Mississippi River were classified as Cyprogenia stegaria (Rafinesque 1820). Previous studies using mitochondrial DNA sequences indicated that C. aberti and C. stegaria were not reciprocally monophyletic groups, suggesting the need for systematic revision. We generated a novel dataset consisting of 10 microsatellite loci and combined it with sequence data from the mitochondrial ND1 gene for 223 Cyprogenia specimens. Bayesian analysis of the ND1 nucleotide sequences identified two divergent clades that differ by 15.9%. Members of these two clades occur sympatrically across most sampling locations. In contrast, microsatellite genotypes support recognition of three allopatric clusters defined by major hydrologic basins. The divergent mitochondrial lineages are highly correlated with the color of the conglutinate lures used by mussels to attract and infest host fishes, and tests for selection at the ND1 locus were positive. We infer that the incongruence between mtDNA and microsatellite data in Cyprogenia may be the result of a combination of incomplete lineage sorting and balancing selection on lure color. Our results provide further evidence that mitochondrial markers are not always neutral with respect to selection, and highlight the potential problems of relying on a single‐locus‐marker for delineating species.     

Life history and production of Paranephrops zealandicus in a forest stream, with comments about the sustainable harvest of a freshwater crayfish

1. Life history and production were assessed for the crayfish Paranephrops zealandicus in three reaches of a headwater stream with a catchment of regenerating coniferous-broadleaf forest in the south-east of the South Island of New Zealand.
2. Crayfish density ranged from 3 to 4 m⁻² in riffles and 4–12 m⁻² in pools, depending on reach. Crayfish biomass (4–33 g AFDW m⁻²) and annual production (2–11 g AFDW m⁻²) were high compared with values reported elsewhere, while P:B ratio was low (0.33–0.43). This substantial production was dependent primarily upon high biomass rather than high growth rate.
3. The crayfish of this population rank amongst the longest lived and slowest growing ever recorded. Individuals estimated to be 16+ year of age were not uncommon. Females became reproductively active at 6+ year. Fewer than 4% of females carried eggs, and young remained attached to females for at least 15 months.
4. We propose that characteristics of this population are the consequence of a cool thermal regime (mean daily stream temperature = 7.0 °C, range 1.8–11.9 °C), and that low biomass turnover and poor reproductive rate precludes any sustainable commercial harvest of crayfish from streams in New Zealand with similar thermal regimes.     

Length‐specific growth rates in freshwater mussels (Bivalvia: Unionidae): extreme longevity or generalized growth cessation?

1. North American lacustrine freshwater mussels (Bivalvia: Unionidae) are one of the world's most imperilled groups of organisms. Knowledge of their age structure and longevity is needed for the understanding and management of mussels. Current methods for age estimation in freshwater mussels are insufficient and may have resulted in an erroneous view of the ages of lacustrine freshwater mussels.
2. We collected growth data through mark-recapture in Minnesota and Rhode Island, U.S.A., examining four lentic populations of three of the most common species of freshwater mussels, Elliptio complanata , Lampsilis siliquoidea , and Pyganodon grandis . Using an inversion of the von Bertalanffy growth equation, we estimated age at length from length-specific growth relationships.
3. In some populations, lacustrine mussels may be much older than previously predicted. Ages predicted from actual growth rates suggest that individuals in some populations frequently reach ages in excess of a century, placing unionid mussels among the Earth's longest-lived animals. Alternatively, if growth has only recently slowed in these populations, generalized growth cessation may be occurring over a broad distributional range of some common North American lacustrine mussels.