Carol A. Stepien

Invasion genetics of Ponto-Caspian gobies in the Great Lakes: a 'cryptic' species, absence of founder effects, and comparative risk analysis.

Genetic variability and structure of nonindigenous vs native populations are compared for the Eurasian round goby Neogobius melanostomus and the tubenose goby Proterorhinus marmoratus, which both invaded Lake St. Clair of the North American Great Lakes about 1990. The round goby spread rapidly to all of the Great Lakes and the tubenose goby largely has been restricted to Lake St. Clair, with some recent range extension into western Lake Erie. Risk analyses may indicate whether genetic variability of colonizers is predictive of their relative invasive and establishment successes. The present investigation examined DNA sequence variation across the left domain of the mitochondrial DNA cytochrome b gene in round and tubenose gobies from Eurasian and Great Lakes locations. We also sequenced six additional Neogobius species (including the monkey N.␣fluviatilis, racer N. gymnotrachelus, and bigheadN. kessleri gobies that have been ‘on the move’ in Europe) and the knout goby Mesogobius batrachocephalus from the Black Sea in order to develop diagnostic genetic characters to identify them in case of future and/or undetected invasions and to delineate their phylogenetic relationships. Results show that a diverse number of haplotypes characterize round and tubenose goby populations from both North America and Eurasian sites, fitting a risk analysis prediction of high genetic variability in their successful introductions. Phylogenetic results indicate that the current genus Neogobius is paraphyletic and that the subgenusApollonia thus should be elevated to the level of genus, containingApollonia (N.) melanostomus (the round goby) andA. (N.) fluviatilis (the monkey goby). In addition, there appear to be two separate species of Proterorhinus marmoratus, a marine P. marmoratus Pallas 1814 in the Black Sea (matching the original type locality), and a ‘cryptic’ freshwater species in the Danube and Dnieper Rivers and probably other Eurasian freshwater habitats, as well as invasive in the Great Lakes. We suggest resurrecting the name P. semilunaris Heckel 1837 for the freshwater species (a taxon that was originally described from rivers draining into the Aegean Sea and the Danube River, but was later placed in synonymy with P. marmoratus).

Invasion genetics of the Eurasian round goby in North America: tracing sources and spread patterns

The Eurasian round goby Neogobius melanostomus (Apollonia melanostoma) invaded the North American Great Lakes in 1990 through ballast water, spread rapidly, and now is widely distributed and moving through adjacent tributaries. We analyse its genetic diversity and divergence patterns among 25 North American (N = 744) and 22 Eurasian (N = 414) locations using mitochondrial DNA cytochrome b gene sequences and seven nuclear microsatellite loci in order to: (i) identify the invasions founding source(s), (ii) test for founder effects, (iii) evaluate whether the invasive range is genetically heterogeneous, and (iv) determine whether fringe and central areas differ in genetic diversity. Tests include FST analogues, neighbour‐joining trees, haplotype networks, Bayesian assignment, Monmonier barrier analysis, and three‐dimensional factorial correspondence analysis. We recovered 13 cytochrome b haplotypes and 232 microsatellite alleles in North America and compared these to variation we previously described across Eurasia. Results show: (i) the southern Dnieper River population was the primary Eurasian donor source for the round gobys invasion of North America, likely supplemented by some alleles from the Dniester and Southern Bug rivers, (ii) the overall invasion has high genetic diversity and experienced no founder effect, (iii) there is significant genetic structuring across North America, and (iv) some expansion areas show reduced numbers of alleles, whereas others appear to reflect secondary colonization. Sampling sites in Lake Hurons Saginaw Bay and Lake Ontario significantly differ from all others, having unique alleles that apparently originated from separate introductions. Substantial genetic variation, multiple founding sources, large number of propagules, and population structure thus likely aided the gobys ecological success.

Signatures of vicariance, postglacial dispersal and spawning philopatry: population genetics of the walleye Sander vitreus

Population genetic relationships reveal the signatures of current processes such as reproductive behaviour and migration, as well as historic events including vicariance and climate change. We analyse population structure of native walleye Sander vitreus across North America, encompassing 10 nuclear DNA microsatellite loci, 26 spawning sites and 921 samples from watersheds across the Great Lakes, Lake Winnipeg, upper Mississippi River, Ohio River and Mobile Bay of the Gulf Coast. Geographical patterning is assessed using phylogenetic trees, pairwise FST analogues, hierarchical partitioning, Mantel regression, Bayesian assignment and Monmonier geographical networks. Results reveal congruent divergences among population groups, corresponding to historic isolation in glacial refugia, dispersal patterns and basin divisions. Broad‐scale relationships show genetic isolation with geographical distance, but reproductive groups within basins do not – with some having pronounced differences. Greatest divergence distinguishes outlying Gulf Coastal and northwest populations, the latter tracing to dispersal from the Missourian refugium to former glacial Lake Agassiz, and basin isolation ∼7000 ya. Genetic barriers in the Great Lakes separate groups in Lakes Superior, Huron’s Georgian Bay, Erie and Ontario, reflecting contributions from Mississippian and Atlantic refugia, and changes in connectivity patterns. Walleye genetic patterns thus reflect vicariance among watersheds and glacial refugia, followed by re‐colonization pathways and changing drainage connections that established modern‐day northern populations, whose separations are maintained through spawning site fidelity. Conservation management practices should preserve genetic identity and unique characters among these divergent walleye populations.

Ancient divisions, recent expansions: phylogeography and population genetics of the round goby Apollonia melanostoma.

During the past two decades, the round goby Apollonia melanostoma (=Neogobius melanostomus) has expanded its range via shipping transport and canals, extending north and west from the Ponto‐Caspian region of Eurasia and to the North American Great Lakes. Exotic populations of the round goby have been very successful in the Baltic Sea and the Great Lakes regions, exerting significant ecological changes. Our study evaluates the population genetic and biogeographical structure of the round goby across its native and nonindigenous ranges, in light of geological history and its expansion pathways. We analyzed seven new nuclear microsatellite loci and mitochondrial DNA cytochrome b gene sequences from 432 individuals in 22 locations. Population structure was tested using FST‐analogs, phylogenetic trees, clustering diagrams, Bayesian assignment tests and nested clade analyses. Results show that native populations in the Black vs. the Caspian Sea basins diverge by 1.4% and c. 350 000 years, corresponding to closure of their prior connections and supporting the taxonomic separation of the Black Sea A. m. melanostoma from the Caspian Sea A. m. affinis. Their within‐basin populations diverge by ~0.4% and 100 000 years. Nonindigenous populations in the Baltic Sea and Danube and Dnieper Rivers trace to separate northern Black Sea origins, whereas the upper Volga River system houses mixed populations of A. m. melanostoma and A. m. affinis. Native populations average twice the genetic diversity of most exotic sites; however, sites in the Volga River system have high diversity due to mixing of the two taxa. Our results highlight how vicariance and anthropogenic disturbances have shaped a rapidly expanding species’ genetic heritage.

Broad- to fine-scale population genetic patterning in the smallmouth bass Micropterus dolomieu across the Laurentian Great Lakes and beyond: an interplay of behaviour and geography.

Analysis of population genetic relationships reveals the signatures of current processes such as spawning behaviour and migration, as well as those of historical events including vicariance and climate change. This study examines these signatures through testing broad‐ to fine‐scale genetic patterns among smallmouth bass Micropterus dolomieu spawning populations across their native Great Lakes range and outgroup areas, with fine‐scale concentration in Lake Erie. Our primary hypotheses include whether genetic patterns result from behavioural and/or geographical isolation, specifically: (i) Are spawning groups in interconnected waterways genetically separable? (ii) What is the degree of isolation across and among lakes, basins, and tributaries? (iii) Do genetic divergences correspond to geographical distances? and (iv) Are historical colonization patterns from glacial refugia retained? Variation at eight nuclear microsatellite DNA loci are analysed for 666 smallmouth bass from 28 locations, including 425 individuals in Lake Erie; as well as Lakes Superior, Huron, and Ontario, and outgroups from the Mississippi, Ohio, St. Lawrence, and Hudson River drainages. Results reveal marked genetic differences among lake and river populations, as well as surprisingly high divergences among closely spaced riverine sites. Results do not fit an isolation‐by‐geographical‐distance prediction for fine‐scale genetic patterns, but show weak correspondence across large geographical scales. Genetic relationships thus are consistent with hypotheses regarding divergent origins through vicariance in glacial refugia, followed by colonization pathways establishing modern‐day Great Lakes populations, and maintenance through behavioural site fidelity. Conservation management practices thus should preserve genetic identity and unique characters among smallmouth bass populations.

Evolution and biogeography of an emerging quasispecies: diversity patterns of the fish Viral Hemorrhagic Septicemia virus (VHSv).

Viral Hemorrhagic Septicemia virus (VHSv) is an RNA rhabdovirus that causes one of the most important finfish diseases, affecting over 70 marine and freshwater species. It was discovered in European cultured fish in 1938 and since has been described across the Northern Hemisphere. Four strains and several substrains have been hypothesized, whose phylogenetic relationships and evolutionary radiation are evaluated here in light of a quasispecies model, including an in-depth analysis of the novel and especially virulent new substrain (IVb) that first appeared in the North American Laurentian Great Lakes in 2003. We analyze the evolutionary patterns, genetic diversity, and biogeography of VHSv using all available RNA sequences from the glycoprotein (G), nucleoprotein (N), and non-virion (Nv) genes, with Maximum Likelihood and bayesian approaches. Results indicate that the G gene evolves at an estimated rate of μ=2.58×10(-4) nucleotide substitutions per site per year, the N gene at μ=4.26×10(-4), and Nv fastest at μ=1.25×10(-3). Phylogenetic trees from the three genes largely are congruent, distinguishing strains I-IV as reciprocally monophyletic with high bootstrap and posterior probability support. VHSv appears to have originated from a marine ancestor in the North Atlantic Ocean, diverging into two primary clades: strain IV in North America (the Northwestern Atlantic Ocean), and strains I-III in the Northeastern Atlantic region (Europe). Strain II may comprise the basal group of the latter clade and diverged in Baltic Sea estuarine waters; strains I and III appear to be sister groups (according to the G and Nv genes), with the former mostly in European freshwaters and the latter in North Sea marine/estuarine waters. Strain IV is differentiated into three monophyletic substrains, with IVa infecting Northeastern Pacific salmonids and many marine fishes (with 44 unique G gene haplotypes), IVb endemic to the freshwater Great Lakes (11 haplotypes), and a newly-designated IVc in marine/estuarine North Atlantic waters (five haplotypes). Two separate substrains independently appeared in the Northwestern Pacific region (Asia) in 1996, with Ib originating from the west and IVa from the east. Our results depict an evolutionary history of relatively rapid population diversifications in star-like patterns, following a quasispecies model. This study provides a baseline for future tracking of VHSv spread and interpreting its evolutionary diversification pathways.

Metazoan Parasites of Introduced Round and Tubenose Gobies in the Great Lakes: Support for the “Enemy Release Hypothesis”

ABSTRACT Recent invasion theory has hypothesized that newly established exotic species may initially be free of their native parasites, augmenting their population success. Others have hypothesized that invaders may introduce exotic parasites to native species and/or may become hosts to native parasites in their new habitats. Our study analyzed the parasites of two exotic Eurasian gobies that were detected in the Great Lakes in 1990: the round goby Apollonia melanostoma and the tubenose goby Proterorhinus semilunaris. We compared our results from the central region of their introduced ranges in Lakes Huron, St. Clair, and Erie with other studies in the Great Lakes over the past decade, as well as Eurasian native and nonindigenous habitats. Results showed that goby-specific metazoan parasites were absent in the Great Lakes, and all but one species were represented only as larvae, suggesting that adult parasites presently are poorly-adapted to the new gobies as hosts. Seven parasitic species are known to infest the tubenose goby in the Great Lakes, including our new finding of the acanthocephalan Southwellina hispida, and all are rare. We provide the first findings of four parasite species in the round goby and clarified two others, totaling 22 in the Great Lakes—with most being rare. In contrast, 72 round goby parasites occur in the Black Sea region. Trematodes are the most common parasitic group of the round goby in the Great Lakes, as in their native Black Sea range and Baltic Sea introduction. Holarctic trematode Diplostomum spathaceum larvae, which are one of two widely distributed species shared with Eurasia, were found in round goby eyes from all Great Lakes localities except Lake Huron proper. Our study and others reveal no overall increases in parasitism of the invasive gobies over the past decade after their establishment in the Great Lakes. In conclusion, the parasite “load” on the invasive gobies appears relatively low in comparison with their native habitats, lending support to the “enemy release hypothesis.”

Expansion of tubenose gobies Proterorhinus semilunaris into western Lake Erie and potential effects on native species

The Eurasian freshwater tubenose goby Proterorhinus semilunaris (formerly Proterorhinus marmoratus) invaded the Laurentian Great Lakes in the 1990s, presumably via ballast water from transoceanic cargo ships. Tubenose gobies spread throughout Lake St. Clair, its tributaries, and the Detroit River system, and also are present in the Duluth-Superior harbor of Lake Superior. Using seines and bottom trawls, we collected 113 tubenose gobies between July 2007 and August 2009 at several locations in western Lake Erie. The number and range of sizes of specimens collected suggest that that tubenose gobies have become established and self-sustaining in the western basin of Lake Erie. Tubenose gobies reached maximum densities in sheltered areas with abundant macrophyte growth, which also is their common habitat in native northern Black Sea populations. The diet of tubenose gobies was almost exclusively invertebrates, suggesting dietary overlap with other benthic fishes, such as darters (Etheostoma spp. and Percina sp.), madtoms (Noturus spp.), and sculpins (Cottus spp.). A single mitochondrial DNA haplotype was identified, which is the most common haplotype found in the original colonization area in the Lake St. Clair region, suggesting a founder effect. Tubenose gobies, like round gobies Neogobius melanostomus, have early life stages that drift owing to vertical migration, which probably allowed them to spread from areas of colonization. The Lake St. Clair-Lake Erie corridor appears to have served as an avenue for them to spread to the western basin of Lake Erie, and abundance of shallow macrophyte-rich habitats may be a key factor facilitating their further expansion within Lake Erie and the remainder of the Laurentian Great Lakes.

Population genetic diversity and phylogeographic divergence patterns of the yellow perch (Perca flavescens)

ABSTRACT Great Lakes populations of yellow perch have fluctuated throughout past decades to the present due to unstable recruitment patterns and exploitation. Our study analyzes genetic diversity and structure across the native range in order to interpret phylogeographic history and contemporary patterns. We compare complete mitochondrial DNA control region sequence) from 568 spawning individcing all 5 Great Lakes and outlying watersheds from the upper Mississippi River, Lake Winnipeg, Lake Champlain, and Atlantic and Gulf coastal relict populations. Theadces additionally are compared with fine-scale patterns from 334 individuals at 16 spawning across Lake Eries 4 fishery management units. We identify 21 mtDNA haplotypes, including a widespread type that totals 87% of individuals across the Great Lakes. Overall genetic diversity is relatively low in comparison with other Great Lakes fishes, congruent with prior allozyme and microsatellite studies. The largest genetic demarcation separates 2 primary population groups: one in the Great Lakes, Lake Winnipeg, and upper Mississippi River watersheds and the other along the Atlantic and Gulf coasts, together with Lake Champlain; which diverged ∼ 365,000 years ago. In addition, the watersheds house genetically separable groups, whose patterns reflect broad-scale isolation by geographic distance. A few spawning groups show some fine-scale differentiation within Lake Erie, which do not reflect fishery management units and need further study with higherresolution markers.

Patterns of Gene Flow and Genetic Divergence in the Northeastern Pacific Clinidae (Teleostei: Blennioidei), Based on Allozyme and Morphological Data

Genetic relationships and distribution patterns among populations, subspecies, and species of northeastern Pacific myxodin clinids were analyzed from allozyme data. The most recent revision recognized six species and 12 subspecies in two genera, Heterostichus and Gibbonsia. Allozymes from 40 gene loci from all 12 nominal taxa were analyzed to compare heterozygosity levels, HardyWeinberg equilibrium conformance, genetic distances, and phylogenetic relationships. Sample sites ranged from Carmel, California, to central Baja California, Mexico, and included areas of sympatry, disjunct distribution, and relative isolation. Offshore island sites included the California Channel, San Benito, and Guadalupe islands; the last with several endemic nominal taxa. In addition, morphological characters putatively defining closely related taxa were reexamined because preliminary data suggested that some taxonomic separations had been made on the basis of sexually dimorphic characters and ecophenotypic variation.