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Nathan A. Johnson

Publications and source records attributed to Nathan A. Johnson.

At least 19 recordsLinked to original sources

U.S. Geological Survey science vision for native freshwater mussel research in the United States

Executive Summary North America is a global center for native freshwater mussel (order Unionida, hereinafter “mussels”) diversity, with more than 350 species. Mussels are among the most imperiled fauna on the planet. Reasons for both local and widespread declines in mussels are mostly unknown, although the threats may include habitat loss and fragmentation, diseases, environmental contaminants, altered flow regimes, migration barriers to larval hosts, non-native species, and climate change. Over the past three decades, research on mussels has been substantial. Nevertheless, current conservation and management efforts are limited by significant information gaps. For example, the effects of emerging stressors on mussels are largely unknown and identifying when habitats are rehabilitated and suitable for reestablishment of mussels remains challenging. Additionally, historical and current information on the distribution, taxonomy, and life histories are often unreliable or lacking altogether, and more reliable information is needed for many species. We identified focal research themes, goals, and objectives where research on mussels is needed based on information gaps identified through conversations with resource partners across local, regional, and national organizations. Research on biodiversity seeks to enhance the diversity of mussel species and populations to support healthy aquatic ecosystems. Research on emerging stressors seeks to improve the understanding of how mussel species, populations, and communities respond to emerging stressors, including environmental contaminants and climate change. Research on conservation seeks to enhance the recovery of species and populations and to identify data gaps limiting the conservation of mussels and their habitats. Mussels are in urgent need of proactive conservation because they are an integral part of our natural heritage, enhance biodiversity, and provide vital ecological services that support freshwater ecosystems. The U.S. Geological Survey (USGS) has been, and continues to be, a leader in mussel research. Although the USGS is well suited to address the broad-scale multidisciplinary research needed to conserve mussels, the USGS has had substantial loss of scientists with mussel expertise over the past 20 years. However, the breadth of the USGS expertise on mussels can be leveraged internally across other USGS mission and program areas and externally across research partners. Given the breadth and scope of the issues facing mussels across the United States, the research themes outlined in this science vision can only be accomplished through extensive collaborations between the USGS and the full spectrum of natural resource partners, including other Federal and State agencies, Tribal organizations, universities, industries, and nongovernmental organizations.

Circular

Using a multi-model ensemble approach to determine biodiversity hotspots with limited occurrence data in understudied areas: An example using freshwater mussels in México

Species distribution models (SDMs) are an increasingly important tool for conservation particularly for difficult-to-study locations and with understudied fauna. Our aims were to (1) use SDMs and ensemble SDMs to predict the distribution of freshwater mussels in the Pánuco River Basin in Central México; (2) determine habitat factors shaping freshwater mussel occurrence; and (3) use predicted occupancy across a range of taxa to identify freshwater mussel biodiversity hotspots to guide conservation and management. In the Pánuco River Basin, we modeled the distributions of 11 freshwater mussel species using an ensemble approach, wherein multiple SDM methodologies were combined to create a single ensemble map of predicted occupancy. A total of 621 species-specific observations at 87 sites were used to create species-specific ensembles. These predictive species ensembles were then combined to create local diversity hotspot maps. Precipitation during the warmest quarter, elevation, and mean temperature were consistently the most important discriminatory environmental variables among species, whereas land use had limited influence across all taxa. To the best of our knowledge, our study is the first freshwater mussel-focused research to use an ensemble approach to determine species distribution and predict biodiversity hotspots. Our study can be used to guide not only current conservation efforts but also prioritize areas for future conservation and study.

Pánuco

Genetic and morphological characterization of the freshwater mussel clubshell species complex (Pleurobema clava and Pleurobema oviforme) to inform conservation planning

The shell morphologies of the freshwater mussel species Pleurobema clava (federally endangered) and Pleurobema oviforme (species of concern) are similar, causing considerable taxonomic confusion between the two species over the last 100 years. While P. clava was historically widespread throughout the Ohio River basin and tributaries to the lower Laurentian Great Lakes, P. oviforme was confined to the Tennessee and the upper Cumberland River basins. We used two mitochondrial DNA (mtDNA) genes, 13 novel nuclear DNA microsatellite markers, and shell morphometrics to help resolve this taxonomic confusion. Evidence for a single species was apparent in phylogenetic analyses of each mtDNA gene, revealing monophyletic relationships with minimal differentiation and shared haplotypes. Analyses of microsatellites showed significant genetic structuring, with four main genetic clusters detected, respectively, in the upper Ohio River basin, the lower Ohio River and Great Lakes, and upper Tennessee River basin, and a fourth genetic cluster, which included geographically intermediate populations in the Ohio and Tennessee river basins. While principal components analysis (PCA) of morphometric variables (i.e., length, height, width, and weight) showed significant differences in shell shape, only 3% of the variance in shell shape was explained by nominal species. Using Linear Discriminant and Random Forest (RF) analyses, correct classification rates for the two species' shell forms were 65.5% and 83.2%, respectively. Random Forest classification rates for some populations were higher; for example, for North Fork Holston (HOLS), it was >90%. While nuclear DNA and shell morphology indicate that the HOLS population is strongly differentiated, perhaps indicative of cryptic biodiversity, we consider the presence of a single widespread species the most likely biological scenario for many of the investigated populations based on our mtDNA dataset. However, additional sampling of P. oviforme populations at nuclear loci is needed to corroborate this finding.

Indiana, Kentucky, Ohio, Pennsylvania, Tennessee,

Rediscovery and genetic confirmation of the Threeridge Mussel, Amblema plicata (Say, 1817) (Bivalvia, Unionidae), in the Choctawhatchee River, Florida, USA

Recent freshwater mussel research has resulted in rediscovery of several species presumed extinct. We report the rediscovery of Amblema plicata (Say, 1817) in 2019 from the Choctawhatchee River, Florida, USA. Amblema plicata has not been reported in the Choctawhatchee river basin since 1958, more than 61 years ago. This species was collected during the long-term monitoring of freshwater mussels in Florida streams. We provide genetic confirmation of our voucher identification using a DNA barcoding approach and discuss potential risks to A. plicata populations in the Choctawhatchee river basin.

Florida

Genetic analysis of red lionfish Pterois volitans from Florida, USA, leads to alternative North Atlantic introduction scenarios

The red lionfish Pterois volitans is a successful invasive predator across the western North Atlantic, Caribbean, and Gulf of Mexico. The southeast coast of Florida (USA) has been identified as the original introduction location, but genetic analyses including Florida lionfish have yet to investigate introduction scenarios. Here, we assessed the potential lionfish invasion pathways using 1795 sequences from previously published mitochondrial D-loop sequences (n = 1558) and new samples (n = 237) from 6 locations: The Bahamas, Florida Keys, northwest Florida, North Carolina, Panamá, and southeast Florida. None of the assessed Florida lionfish (n = 394) contained the H05-H09 D-loop haplotypes found in The Bahamas, North Carolina, and Bermuda (the Northern Region), indicating that Florida was not the source for these haplotypes. Assessing the mitochondrial population structure, the Florida east coast lionfish grouped with the Caribbean/Gulf of Mexico, as opposed to the Northern Region. To further explore connectivity and invasion pathways, 14 nuclear microsatellite loci were multiplexed on lionfish collected from 15 locations (n = 394). As found in other nuclear lionfish studies, the analyses identified a lack of population structure likely due to founding effects and/or inbreeding in aquaculture brood stocks. Together, the significant haplotype differences and H01-H04 haplotypes refute Florida as the sole source of red lionfish introduction. The results of this study support alternative invasion scenarios, in which Florida was colonized as a secondary introduction site or by individuals from the Northern Region. Understanding invasive species’ population boundaries and dispersal patterns informs local control efforts and management planning for future invasive species introductions.

Florida

Novel molecular resources to facilitate future genetics research on freshwater mussels (Bivalvia: Unionidae)

Molecular data have been an integral tool in the resolution of the evolutionary relationships and systematics of freshwater mussels, despite the limited number of nuclear markers available for Sanger sequencing. To facilitate future studies, we evaluated the phylogenetic informativeness of loci from the recently published anchored hybrid enrichment (AHE) probe set Unioverse and developed novel Sanger primer sets to amplify two protein-coding nuclear loci with high net phylogenetic informativeness scores: fem-1 homolog C (FEM1) and UbiA prenyltransferase domain-containing protein 1 (UbiA). We report the methods used for marker development, along with the primer sequences and optimized PCR and thermal cycling conditions. To demonstrate the utility of these markers, we provide haplotype networks, DNA alignments, and summary statistics regarding the sequence variation for the two protein-coding nuclear loci (FEM1 and UbiA). Additionally, we compare the DNA sequence variation of FEM1 and UbiA to three loci commonly used in freshwater mussel genetic studies: the mitochondrial genes cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase subunit 1 (ND1), and the nuclear internal transcribed spacer 1 (ITS1). All five loci distinguish among the three focal species (Potamilus fragilis, Potamilus inflatus, and Potamilus purpuratus), and the sequence variation was highest for ND1, followed by CO1, ITS1, UbiA, and FEM1, respectively. The newly developed Sanger PCR primers and methodologies for extracting additional loci from AHE probe sets have great potential to facilitate molecular investigations targeting supraspecific relationships in freshwater mussels, but may be of limited utility at shallow taxonomic scales.

Data

The utility of zooarchaeological data to guide listing efforts for an imperiled mussel species (Bivalvia: Unionidae: Pleurobema riddellii)

The status of species in freshwater systems shift over time due to natural and anthropogenic causes. Determining the magnitude and cause of these shifts requires a long-term perspective. This process is complicated when there are also questions about the taxonomic validity of a species. Addressing these issues is important because both can undermine conservation and management efforts if incorrect. Pleurobema riddellii , Louisiana Pigtoe, is under review for protection under the U.S. Endangered Species Act, but its status in the Trinity River basin, where the taxon was described, remains in doubt due to questions about its taxonomy and occurrence within this basin. To address these questions, we compared shell morphometrics of P. riddellii dating to the late Holocene with modern P. riddellii , late Holocene Fusconaia sp., and modern Fusconaia sp. using multivariate analyses to test associations between the putative morphotypes. Based on these analyses, we demonstrate that P. riddellii was likely present in the Trinity during the late Holocene, which indicates questions about its taxonomic validity or presence in this basin are unfounded. Our study further highlights the role zooarchaeological studies can play in status assessments and their utility in better understanding biogeographic patterns for rare species.

Texas

A comparative phylogeographic approach to facilitate recovery of an imperiled freshwater mussel (Bivalvia: Unionida: Potamilus inflatus)

North American freshwaters are among the world’s most threatened ecosystems, and freshwater mussels are among the most imperiled inhabiting these systems. A critical aspect of conservation biology is delineating patterns of genetic diversity, which can be difficult when a taxon has been extirpated from a significant portion of its historical range. In such cases, evaluating conservation and recovery options may benefit by using surrogate species as proxies when assessing overall patterns of genetic diversity. Here, we integrate the premise of surrogate species into a comparative phylogeographic framework to hypothesize genetic relationships between extant and extirpated populations of Potamilus inflatus by characterizing genetic structure in co-distributed congeners with similar life histories and dispersal capabilities. Our mitochondrial and nuclear sequence data exhibited variable patterns of genetic divergence between Potamilus spp. native to the Mobile and Pascagoula + Pearl + Pontchartrain (PPP) provinces. However, hierarchical Approximate Bayesian Computation indicated that the diversification between Mobile and PPP clades was synchronous and represents a genetic signature of a common history of vicariance. Recent fluctuations in sea-level appear to have caused Potamilus spp. in the PPP to form a single genetic cluster, providing justification for using individuals from the Amite River as a source of brood stock to re-establish extirpated populations of P. inflatus. Future studies utilizing eDNA and genome-wide molecular data are essential to better understand the distribution of P. inflatus and establish robust recovery plans. Given the imperilment status of freshwater mussels globally, our study represents a useful methodology for predicting relationships among extant and extirpated populations of imperiled species.

Florida, Alabama, Mississippi, Louisiana

Comparative phylogenomics reveal complex evolution of life history strategies in a clade of bivalves with parasitic larvae (Bivalvia: Unionoida: Ambleminae)

Freshwater mussels are a species‐rich group with biodiversity patterns strongly shaped by a life history strategy that includes an obligate parasitic larval stage. In this study, we set out to reconstruct the life history evolution and systematics in a clade of freshwater mussels adapted to parasitizing a molluscivorous host fish. Anchored hybrid enrichment and ancestral character reconstruction revealed a complex pattern of life history evolution with host switching and multiple instances of convergence, including reduction in size of larvae, increased fecundity, and growth during encapsulation. Our phylogenomic analyses also recovered non‐monophyly of taxa exhibiting multiple traits used as the basis for previous taxonomic hypotheses. Taxa with axe‐head shaped glochidia were resolved as paraphyletic, but our results strongly suggest the complex morphology is an adaptation to reduce larval size, with reduction in size further accentuated in taxa previously assigned to Leptodea . To more accurately reflect the evolutionary history of this group, we make multiple systematic changes, including the description of a new genus, Atlanticoncha gen. nov. , and the synonymy of the genus Leptodea under Potamilus . Our findings contribute to the growing body of literature showing that cladistic hypotheses based solely on morphological characters, including larval morphology, can be flawed in freshwater mussels.

Cladistics

Putative mitochondrial sex determination in the Bivalvia: Insights from a hybrid transcriptome assembly in freshwater mussels

Bivalves exhibit an astonishing diversity of sexual systems, with genetic and environmental determinants of sex, and possibly the only example of mitochondrial genes influencing sex determination pathways in animals. In contrast to all other animal species in which strict maternal inheritance (SMI) of mitochondria is the rule, bivalves possess a system known as doubly uniparental inheritance (DUI) of mitochondria in which maternal and paternal mitochondria (and their corresponding female-transmitted or F mtDNA and male-transmitted or M mtDNA genomes) are transmitted within a species. Species with DUI also possess sex-associated mtDNA-encoded proteins (in addition to the typical set of 13), which have been hypothesized to play a role in sex determination. In this study, we analyzed the sex-biased transcriptome in gonads of two closely-related freshwater mussel species with different reproductive and mitochondrial transmission modes: the gonochoric, DUI species, Utterbackia peninsularis , and the hermaphroditic, SMI species, Utterbackia imbecillis . Through comparative analysis with other DUI and non-DUI bivalve transcriptomes already available, we identify common male and female-specific genes, as well as SMI and DUI-related genes, that are probably involved in sex determination and mitochondrial inheritance in this animal group. Our results contribute to the understanding of what could be the first animal sex determination system involving the mitochondrial genome

Florida

Research priorities for freshwater mussel conservation assessment

Freshwater mussels are declining globally, and effective conservation requires prioritizing research and actions to identify and mitigate threats impacting mussel species. Conservation priorities vary widely, ranging from preventing imminent extinction to maintaining abundant populations. Here, we develop a portfolio of priority research topics for freshwater mussel conservation assessment. To address these topics, we group research priorities into two categories: intrinsic or extrinsic factors. Intrinsic factors are indicators of organismal or population status, while extrinsic factors encompass environmental variables and threats. An understanding of intrinsic factors is useful in monitoring, and of extrinsic factors are important to understand ongoing and potential impacts on conservation status. This dual approach can guide conservation status assessments prior to the establishment of priority species and implementation of conservation management actions.

Biological Conservation

Integrative taxonomy resolves taxonomic uncertainty for freshwater mussels being considered for protection under the U.S. Endangered Species Act

Objectively delimiting species boundaries remains an important challenge in systematics and becomes urgent when unresolved taxonomy complicates conservation and recovery efforts. We examined species boundaries in the imperiled freshwater mussel genus Cyclonaias (Bivalvia: Unionidae) using morphometrics, molecular phylogenetics, and multispecies coalescent models to help guide pending conservation assessments and legislative decisions. Congruence across multiple lines of evidence indicated that current taxonomy overestimates diversity in the C. pustulosa species complex. The only genetically and morphologically diagnosable species in the C. pustulosa species complex were C. pustulosa and C. succissa and we consider C. aurea , C. houstonensis, C. mortoni , and C. refulgens to be synonyms of C. pustulosa . In contrast, all three species in the C. nodulata complex ( C. necki , C. nodulata , and C. petrina ) were genetically, geographically, and morphologically diagnosable. Our findings have important conservation and management implications, as three nominal species ( C. aurea , C. houstonensis , and C. petrina ) are being considered for protection under the Endangered Species Act.

Scientific Reports

Cytonuclear discordance in the Florida Everglades invasive Burmese python (Python bivittatus) population reveals possible hybridization with the Indian python (P. molurus)

The invasive Burmese python ( Python bivittatus ) has been reproducing in the Florida Everglades since the 1980s. These giant constrictor snakes have caused a precipitous decline in small mammal populations in southern Florida following escapes or releases from the commercial pet trade. To better understand the invasion pathway and genetic composition of the population, two mitochondrial (mtDNA) loci across 1,398 base pairs were sequenced on 426 snakes and 22 microsatellites were assessed on 389 snakes. Concatenated mtDNA sequences produced six haplotypes with an average nucleotide and haplotype diversity of π = 0.002 and h = 0.097, respectively. Samples collected in Florida from morphologically identified P. bivittatus snakes were similar to published cytochrome oxidase 1 and cytochrome b sequences from both P. bivittatus and Python molurus and were highly divergent (genetic distances of 5.4% and 4.3%, respectively). The average number of microsatellite alleles and expected heterozygosity were N A = 5.50 and H E = 0.60, respectively. Nuclear Bayesian assignment tests supported two genetically distinct groups and an admixed group, not geographically differentiated. The effective population size ( N E = 315.1) was lower than expected for a population this large, but reflected the low genetic diversity overall. The patterns of genetic diversity between mtDNA and microsatellites were disparate, indicating nuclear introgression of separate mtDNA lineages corresponding to cytonuclear discordance. The introgression likely occurred prior to the invasion, but genetic information on the native range and commercial trade is needed for verification. Our finding that the Florida python population is comprised of distinct lineages suggests greater standing variation for adaptation and the potential for broader areas of suitable habitat in the invaded range.

Florida

Molecular and morphometric analyses reveal cryptic diversity within freshwater mussels (Bivalvia: Unionidae) of the western Gulf coastal drainages of the USA

Past geological processes and climate change affected current species distributions and the genetic structure of riverine fauna. Western Gulf of Mexico coastal rivers harbour four mussel species within the genus Fusconaia (Bivalvia: Unionida). The distributions of these species are unclear owing to their indistinguishable shell morphologies. Using molecular phylogenetic and Fourier morphometric analyses, we examined phylogenetic relationships and morphological variation among the species and made inferences about the role of past geological and climatic factors in shaping the current genetic structure and distributions of these species in the region. Our results showed the presence of three Fusconaia species within the region: Fusconaia askewi , Fusconaia chunii and Fusconaia flava . We confirmed that Fusconaia lananensis is a junior synonym of F. askewi and that F. chunii is genetically distinct from F. askewi . The Trinity River has syntopic F. flava whose morphologies are indistinguishable from those of F. chunii . Divergence-time estimates matched major geological and climatic events in the region, where climate-driven river formations during the mid-Miocene to Pleistocene caused major diversification of Fusconaia species. Knowledge gained from the present study provides a better understanding of vicariant events that shaped current species distributions and helps to identify conservation priorities that apply to the Fusconaia species.

Biological Journal of the Linnean Society

Cumulative spring discharge and survey effort influence occupancy and detection of a threatened freshwater mussel, the Suwannee Moccasinshell

Freshwater mussels (Unionidae) are among the most imperiled groups of organisms in the world, and the lack of information regarding species distributions, life-history characteristics, and ecological and biological requirements may limit the protection of remaining mussel populations. We examined the influence of hydrologic factors on the occurrence of the Suwannee Moccasinshell Medionidus walkeri, a federally threatened freshwater mussel species, endemic to the Suwannee River Basin in Georgia and Florida. We also evaluated the influence of survey effort on detection of Suwannee Moccasinshell during field surveys. We compiled all recent (2013–2016) mussel survey records in the Suwannee River Basin. We calculated cumulative discharge contributed by upstream springs for each of 220 survey locations. We combined the spring discharge predictor variable with Suwannee Moccasinshell detection and nondetection data from each survey location to develop a suite of occupancy models. Modeling results indicated that detection of Suwannee Moccasinshell during surveys was strongly and positively related to survey effort. Modeling results also indicated that sites with cumulative spring discharge inputs exceeding ∼28 cubic meters per second were most likely (i.e., predicted occupancy probabilities >0.5) to support Suwannee Moccasinshell populations. However, occupancy declined in the lowermost reaches of the Suwannee mainstem despite high spring discharge inputs, presumably due to greater tidal influences and differences in physicochemical habitat conditions. Historical localities where Suwannee Moccasinshell has presumably been extirpated are all devoid of springs in their upstream watersheds. We hypothesize that springs may buffer extremely tannic, and at times polluted, surface waters, in addition to maintaining adequate flows during periods of drought, thereby promoting the persistence of Suwannee Moccasinshell populations. Our study suggests that springs are a critical resource for Suwannee Moccasinshell and may be more important for conservation planning than was previously recognized.

Florida

Deciphering the link between doubly uniparental inheritance of mtDNA and sex determination in bivalves: Clues from comparative transcriptomics

Bivalves exhibit an astonishing diversity of sexual systems and sex-determining mechanisms. They can be gonochoric, hermaphroditic or androgenetic, with both genetic and environmental factors known to determine or influence sex. One unique sex-determining system involving the mitochondrial genome has also been hypothesized to exist in bivalves with doubly uniparental inheritance (DUI) of mtDNA. However, the link between DUI and sex determination remains obscure. In this study, we performed a comparative gonad transcriptomics analysis for two DUI-possessing freshwater mussel species to better understand the mechanisms underlying sex determination and DUI in these bivalves. We used a BLAST reciprocal analysis to identify orthologs between Venustaconcha ellipsiformis and Utterbackia peninsularis and compared our results with previously published sex-specific bivalve transcriptomes to identify conserved sex-determining genes. We also compared our data with other DUI species to identify candidate genes possibly involved in the regulation of DUI. A total of ∼12,000 orthologous relationships were found, with 2,583 genes differentially expressed in both species. Among these genes, key sex-determining factors previously reported in vertebrates and in bivalves (e.g., Sry, Dmrt1, Foxl2 ) were identified, suggesting that some steps of the sex-determination pathway may be deeply conserved in metazoans. Our results also support the hypothesis that a modified ubiquitination mechanism could be responsible for the retention of the paternal mtDNA in male bivalves, and revealed that DNA methylation could also be involved in the regulation of DUI. Globally, our results suggest that sets of genes associated with sex determination and DUI are similar in distantly-related DUI species.

Genome Biology and Evolution

A semi-arid river in distress: Contributing factors and recovery solutions for three imperiled freshwater mussels (Family Unionidae) endemic to the Rio Grande basin in North America

Freshwater resources in arid and semi-arid regions are in extreme demand, which creates conflicts between needs of humans and aquatic ecosystems. The Rio Grande basin in the southwestern United States and northern Mexico exemplifies this issue, as much of its aquatic biodiversity is in peril as a result of human activities. Unionid mussels have been disproportionately impacted, though the specific factors responsible for their decline remain largely unknown. This is problematic because the Rio Grande basin harbors one federally endangered unionid mussel ( Popenaias popeii , Texas Hornshell) plus two other mussel species ( Potamilus metnecktayi , Salina Mucket; and Truncilla cognata , Mexican Fawnsfoot), which are also being considered for listing under the U.S. Endangered Species Act. To date, surveys for these species have not corrected for variability in detection so current range estimates may be inaccurate. Using single occupancy-modeling to estimate detection and occupancy at 115 sites along ~800 river kilometers of the Rio Grande in Texas, we found that detection probabilities were relatively high, indicating that our survey design was efficient. In contrast, the estimated occupancy was low, indicating that our focal species were likely rare within the Rio Grande drainage. In general, the predicted occupancy of our focal species was low throughout their respective ranges, indicating possible range declines. A comparison of currently occupied ranges to presumptive ranges underscores this point. The best-approximating models indicated that occupancy was influenced by habitat, water quantity and quality, and proximity to large-scale human activities, such as dams and major urban centers. We also discuss a series of conservation options that may not only improve the long-term prognosis of our focal species but also other aquatic taxa.

Rio Grande basin

Molecular and morphological data reveal non-monophyly and speciation in imperiled freshwater mussels (Anodontoides and Strophitus)

Accurate taxonomic placement is vital to conservation efforts considering many intrinsic biological characteristics of understudied species are inferred from closely related taxa. The rayed creekshell, Anodontoides radiatus (Conrad, 1834), exists in the Gulf of Mexico drainages from western Florida to Louisiana and has been petitioned for listing under the Endangered Species Act. We set out to resolve the evolutionary history of A. radiatus, primarily generic placement and species boundaries, using phylogenetic, morphometric, and geographic information. Our molecular matrix contained 3 loci: cytochrome c oxidase subunit I, NADH dehydrogenase subunit I, and the nuclear-encoded ribosomal internal transcribed spacer I. We employed maximum likelihood and Bayesian inference to estimate a phylogeny and test the monophyly of Anodontoides and Strophitus. We implemented two coalescent-based species delimitation models to test seven species models and evaluate species boundaries within A. radiatus. Concomitant to molecular data, we also employed linear morphometrics and geographic information to further evaluate species boundaries. Molecular and morphological evidence supports the inclusion of A. radiatus in the genus Strophitus, and we resurrect the binomial Strophitus radiatus to reflect their shared common ancestry. We also found strong support for polyphyly in Strophitus and advocate the resurrection of the genus Pseudodontoideus to represent ‘Strophitus’ connasaugaensis and ‘Strophitus’ subvexus. Strophitus radiatus exists in six well-supported clades that were distinguished as evolutionary independent lineages using Bayesian inference, maximum likelihood, and coalescent-based species delimitation models. Our integrative approach found evidence for as many as 4 evolutionary divergent clades within S. radiatus. Therefore, we formally describe two new species from the S. radiatus species complex (Strophitus williamsi and Strophitus pascagoulaensis) and recognize the potential for a third putative species (Strophitus sp. cf. pascagoulaensis). Our findings aid stakeholders in establishing conservation and management strategies for the members of Anodontoides, Strophitus, and Pseudodontoideus.

Alabama, Florida, Louisiana, Mississippi