Seasonal resource selection by blue suckers Cycleptus elongatus

Telemetry data were used to quantify seasonal resource selection and rank‐preferred habitat types for blue suckers Cycleptus elongatus in the middle Missouri River and its tributaries, Iowa, Nebraska, South Dakota, U.S.A. Habitat diversity required by C. elongatus to complete its annual life‐history cycle was evident by interseasonal variation in habitat selection. Tributaries and unchannelized portions of the middle Missouri River system were selected during the inferred spawning season (spring), habitats along shorelines in the channelized middle Missouri River were selected for postspawning (summer) and areas of reduced current were generally selected for the period preceding spawning (autumn). This study provides insight into mature C. elongatus behaviour throughout its annual life cycle. Furthermore, these results document the interseasonal variability in habitat selection of C. elongatus and highlight the need for development of habitat selection models for other imperilled fish species in aquatic systems affected by anthropogenic developments.     

Genetic variation in westslope cutthroat trout <Emphasis Type="Italic">Oncorhynchus clarkii lewisi</Emphasis>: implications for conservation

Twenty-five populations of westslope cutthroat trout from throughout their native range were genotyped at 20 microsatellite loci to describe the genetic structure of westslope cutthroat trout. The most genetic diversity (heterozygosity, allelic richness, and private alleles) existed in populations from the Snake River drainage, while populations from the Missouri River drainage had the least. Neighbor-joining trees grouped populations according to major river drainages. A great amount of genetic differentiation was present among and within all drainages. Based on Nei’s D _S , populations in the Snake River were the most differentiated, while populations in the Missouri River were the least. This pattern of differentiation is consistent with a history of sequential founding events through which westslope cutthroat trout may have experienced a genetic bottleneck as they colonized each river basin from the Snake to the Clark Fork to the Missouri river. These data should serve as a starting point for a discussion on management units and possible distinct population segments. Given the current threats to the persistence of westslope cutthroat trout, and the substantial genetic differentiation between populations, these topics warrant attention.     

Range‐wide assessment of pallid sturgeon Scaphirhynchus albus (Forbes & Richardson, 1905) relative condition

Pallid sturgeon Scaphirhynchus albus relative condition has been observed to be declining along the Nebraska reach (rkm 1212.6–801.3) of the Missouri River over the past several years; therefore, pallid sturgeon capture data was synthesized from the entire Missouri and Middle Mississippi rivers to document and compare how pallid sturgeon condition varies spatially and temporally throughout much of their current range. The study area was subdivided into four river reaches based on a priori statistical differences for pallid sturgeon catches from 2003 to 2015. Pallid sturgeon in the Middle Mississippi River (Alton Dam [rkm 321.9]) to the confluence of the Ohio River (rkm 0.0) were in the best condition while pallid sturgeon in the Middle Missouri River (Fort Randall Dam [rkm 1416.2]) to the Grand River confluence (rkm 402.3) were in the poorest condition. Furthermore, pallid sturgeon condition in the Upper Missouri River (Fort Peck Dam [rkm 2850.9] to the headwaters of Lake Sakakawea [rkm 2523.5] and lower Yellowstone River) and the Lower Missouri River (Grand River confluence to the Mississippi River confluence [rkm 0.0]) were significantly less than in the Middle Mississippi River but significantly higher than the Middle Missouri River. Temporally, pallid sturgeon condition was highly variable. Relative condition in the Middle Mississippi River was consistently above average (Kn = 1.1). Comparatively, Kn throughout the Missouri River rarely exceeded “normal” (Kn = 1.0), with Kn in the middle and lower reaches of the Missouri River having declined to the lowest observed. As pallid sturgeon recovery efforts continue, understanding the range‐wide differences and effects on condition could be critical, as poor condition may cause maturation delays, reproductive senescence or even mortality, which affects the likelihood of natural reproduction and recruitment.     

Documentation of lake sturgeon (Acipenser fulvescens Rafinesque, 1817) recovery and spawning success from a restored population in the Mississippi River,Missouri, USA

Lake sturgeon Acipenser fulvescens are considered rare and were nearly extirpated in the Mississippi River in Missouri by 1931 as a result of overfishing and habitat fragmentation. Propagation efforts have been implemented by the Missouri Department of Conservation since 1984 as means to restore the lake sturgeon population. Although recent population increases have been observed, a formalized evaluation to determine if lake sturgeon are self‐sustaining in the Missouri portion of the Mississippi River has not been completed. Therefore, the objectives of this study were to: (i) determine the proportion of reproductive individuals, (ii) evaluate seasonal movement patterns of adults, and (iii) validate purported spawning locations within the Mississippi River in Missouri. Lake sturgeon catch data indicated that approximately 11 percent of the population are reproductively mature. Additionally, telemetry data confirms that the greatest movement by adult lake sturgeon occurs during spring, which suggests spawning behavior. Finally, it was possible to document lake sturgeon embryos and emergent fry larvae below Melvin Price Locks and Dam 26 in the Upper Mississippi River near St. Louis, Missouri. Water velocity, depth, and substrate size were measured at this location and embryos were collected and hatched in the laboratory. River gage data suggest that spawning behavior may have been elicited by a large influx of water during a drawdown period of water above the dam. This study represents the first documented spawning of A. fulvescens in the Mississippi River and highlights the success of recovery efforts in Missouri.     

Morphometric variation among river sturgeons (Scaphirhynchus spp.) of the Middle and Lower Mississippi River

Pallid sturgeon (Scaphirhynchus albus) captured in the Middle and Lower Mississippi River (i.e. below St. Louis, MO, USA) are morphologically very similar to shovelnose sturgeon (Scaphirhynchus platorynchus). Available empirical data are limited to a few studies based on low sample sizes from disjointed populations. Geneticists are currently searching for markers that will differentiate the two species, but the need for unequivocal species‐specific field characters remains. Continuation of commercial fishing for shovelnose sturgeon in some states necessitates an immediate means for accurate field identifications. Previous studies of lower basin river sturgeon classified individuals with simple morphometric character indices and interpreted intermediacy as interspecific hybridization. In this study, morphometric variation among Scaphirhynchus specimens from the Middle and Lower Mississippi River is examined for evidence of hybridization. Data are compared for large (>250‐mm standard length) hatchery‐reared and wild pallid specimens and wild shovelnose specimens. Specimens are compared using two morphometric character indices, two morphometric/meristic character indices and principal components analysis. Results indicate substantial morphological variation among pallid sturgeon below the mouth of the Missouri River. The amount of variation appears to decrease downstream in the Mississippi River. Sheared principal components analysis of morphometric data shows complete separation of shovelnose and pallid sturgeon specimens, whereas character indices indicate overlap. Both character indices and sheared principal components analysis demonstrate that pallid sturgeon in the Lower Mississippi River are morphologically more similar to shovelnose sturgeon than are pallids from the Upper Missouri River. This similarity, explained in previous studies as hybridization, may be the result of latitudinal morphometric variation and length‐at‐age differences between populations of the upper and lower extremes of the range.     

SNP discovery in wild and domesticated populations of blue catfish,Ictalurus furcatus,using genotyping‐by‐sequencing and subsequent SNP validation

Blue catfish, Ictalurus furcatus, are valued in the United States as a trophy fishery for their capacity to reach large sizes, sometimes exceeding 45 kg. Additionally, blue catfish × channel catfish (I. punctatus) hybrid food fish production has recently increased the demand for blue catfish broodstock. However, there has been little study of the genetic impacts and interaction of farmed, introduced and stocked populations of blue catfish. We utilized genotyping‐by‐sequencing (GBS) to capture and genotype SNP markers on 190 individuals from five wild and domesticated populations (Mississippi River, Missouri, D&B, Rio Grande and Texas). Stringent filtering of SNP‐calling parameters resulted in 4275 SNP loci represented across all five populations. Population genetics and structure analyses revealed potential shared ancestry and admixture between populations. We utilized the Sequenom MassARRAY to validate two multiplex panels of SNPs selected from the GBS data. Selection criteria included SNPs shared between populations, SNPs specific to populations, number of reads per individual and number of individuals genotyped by GBS. Putative SNPs were validated in the discovery population and in two additional populations not used in the GBS analysis. A total of 64 SNPs were genotyped successfully in 191 individuals from nine populations. Our results should guide the development of highly informative, flexible genotyping multiplexes for blue catfish from the larger GBS SNP set as well as provide an example of a rapid, low‐cost approach to generate and genotype informative marker loci in aquatic species with minimal previous genetic information.     

Otolith trace element and stable isotopic compositions differentiate fishes from the Middle Mississippi River, its tributaries, and floodplain lakes

Naturally occurring stable isotope and trace elemental markers in otoliths have emerged as powerful tools for determining natal origins and environmental history of fishes in a variety of marine and freshwater environments. However, few studies have examined the applicability of this technique in large river-floodplain ecosystems. This study evaluated otolith microchemistry and stable isotopic composition as tools for determining environmental history of fishes in the Middle Mississippi River, its tributaries, and floodplain lakes in Illinois and Missouri, USA. Fishes were collected from 14 sites and water samples obtained from 16 sites during summer and fall 2006 and spring 2007. Otolith and water samples were analyzed for stable oxygen isotopic composition (δ¹⁸O) and concentrations of a suite of trace elements; otoliths were also analyzed for carbon isotopic composition (δ¹³C). Tributaries, floodplain lakes, and the Mississippi and Lower Missouri Rivers possessed distinct isotopic and elemental signatures that were reflected in fish otoliths. Fish from tributaries on the Missouri and Illinois sides of the middle Mississippi River could also be distinguished from one another by their elemental and isotopic fingerprints. Linear discriminant function analysis of otolith chemical signatures indicated that fish could be classified back to their environment of capture (Mississippi River, floodplain lake, tributary on the Illinois or Missouri side of the Mississippi River, or lower Missouri River) with 71–100% accuracy. This study demonstrates the potential applicability of otolith microchemistry and stable isotope analyses to determine natal origins and describe environmental history of fishes in the Middle Mississippi River, its tributaries, and floodplain lakes. The ability to reconstruct environmental history of individual fish using naturally occurring isotopic markers in otoliths may also facilitate efforts to quantify nutrient and energy subsidies to the Mississippi River provided by fishes that emigrate from floodplain lakes or tributaries.     

Phylogeography of Rhinichthys cataractae (Teleostei: Cyprinidae): pre‐glacial colonization across the Continental Divide and Pleistocene diversification within the Rio Grande drainage

The longnose dace, Rhinichthys cataractae, is a primary freshwater fish inhabiting riffle habitats in small headwater rivers and streams across the North American continent, including drainages east and west of the Continental Divide. The mitochondrially encoded cytochrome b gene (1140 bp) and 2298–2346 bp of the nuclear‐encoded genes S7 and RAG1 were obtained from 87 individuals of R. cataractae (collected from 17 sites throughout its range) and from several close relatives. Phylogenetic analyses recovered a monophyletic R. cataractae species‐group that contained Rhinichthys evermanni, Rhinichthys sp. ‘Millicoma dace’, and a non‐exclusive R. cataractae. Within the R. cataractae species‐group, two well‐supported lineages were identified, including a western lineage (containing R. evermanni, R. sp. ‘Millicoma dace’ and individuals of R. cataractae from Pacific slope drainages) and an eastern lineage (containing individuals of R. cataractae from Arctic, Atlantic, and Gulf slope drainages). Within the eastern lineage of R. cataractae, two well‐supported groups were recovered: a south‐eastern group, containing individuals from the Atlantic slope, southern tributaries to the Mississippi River, and the Rio Grande drainage; and a north‐eastern group, containing individuals from the Arctic slope and northern tributaries to the Mississippi River. Estimates of the timing of divergence within the R. cataractae species‐group, combined with ancestral area‐reconstruction methods, indicate a separation between the eastern and western lineages during the Pliocene to early‐Pleistocene, with a direction of colonization from the west of the Continental Divide eastward. Within the southern portion of its range, R. cataractae likely entered the Rio Grande drainage during the Pleistocene via stream capture events between the Arkansas River (Mississippi River drainage) and headwaters of the Rio Grande. A close relationship between populations of R. cataractae in the Rio Grande drainage and the adjacent Canadian River (Mississippi River drainage) is consistent with hypothesized stream capture events between the Pecos (Rio Grande drainage) and Canadian rivers during the late‐Pleistocene. The population of R. cataractae in the lower Rio Grande may have become separated from other populations in the Rio Grande drainage (upper Rio Grande and Pecos River) and Canadian River during the late‐Pleistocene, well before initiation of recent and significant anthropogenic disturbance within the Rio Grande drainage. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 317–333.     

Phylogeography and post-glacial colonization patterns of the rainbow darter, Etheostoma caeruleum (Teleostei: Percidae)

Aim To examine the effects of historical climate change and drainage isolation on the distribution of mitochondrial DNA cytochrome b genetic variation within the rainbow darter, Etheostoma caeruleum (Percidae: Etheostomatinae). Location Eastern North American streams including tributaries to the Mississippi River, Great Lakes, Potomac River and Hudson Bay drainages. Methods Parsimony analyses, Bayesian analyses and haplotype networks of mitochondrial DNA sequences. Results Four major clades were recovered from sampled populations of E. caeruleum. Three of four clades are distributed in the western portion of the species’ range (primarily west of the Mississippi River). Samples from this region do not form a monophyletic group, and sequences often vary greatly between samples from adjacent stream systems (up to 7.2% divergence). A basal clade includes samples from the White River system in the Ozark Highlands. The northern Ozarks–upper Midwest clade includes samples from Missouri River tributaries and the upper Midwest (Hudson Bay, upper Mississippi River, and western Lake Michigan drainage). The eastern clade is composed of individuals from the Ohio River, Great Lakes and Potomac River. The Mississippi River corridor clade includes samples from middle and lower Mississippi River tributaries. Main conclusions The four major clades of E. caeruleum are deep allopatric lineages with well‐defined boundaries and have additional phylogeographical structure within each clade. The Ozark Highlands have the greatest levels of diversity relative to distributional area, with marked cytochrome b subdivisions between adjacent stream systems. Samples from previously glaciated areas do not have a subset of the cytochrome b diversity found in unglaciated areas, but four separate source areas are identified based on phylogenetic analyses. Dispersal into previously glaciated areas followed several known glacial outlets and, based on sequence divergence between populations, may have occurred during different glacial or interglacial stages. The disjunct distribution and cytochrome b pattern of E. caeruleum in the Mississippi River corridor clade is consistent with late Pleistocene and Recent changes in the course and characteristics of the middle and lower Mississippi River. Phylogeographical boundaries between clades of E. caeruleum correspond to independent sources of biogeographical information and provide insight into historical stream drainage relationships, post‐glacial colonization and drainage isolation patterns.     

Assessing phylogeographic variation in the Rosyside Dace (Teleostei,Leuciscidae), a widespread morphologically variable taxon

Clinostomus (Leuciscidae) is a wide-ranging freshwater fish genus that occurs throughout eastern North America and southern portions of Canada with two species currently recognized: C. elongatus and C. funduloides. A previous taxonomic study of C. funduloides recognized two subspecies (estor and funduloides) and one diagnosed, but undescribed subspecies based on morphological characteristics and geographic distribution. In this study, we used three molecular markers (cytochrome b, S7 intron 1 and growth hormone intron 4) to test the three lineage hypothesis and evaluate genetic variation of C. funduloides across the range using Bayesian inference. Our results indicate that C. funduloides is not monophyletic, as individuals of C. elongatus nest within C. funduloides in both the mtDNA and nDNA phylogenetic analyses, although the position of C. elongatus varies between data sets. In addition, some of the recovered clades are deeply divergent from one another, further supporting the distinctiveness of many of the populations. Overall, these results suggest that subspecies designations are not warranted and a taxonomic revision is needed as Clinostomus is likely more diverse than is currently recognized.     

Stock structure of shovelnose sturgeon analyzed with microsatellite DNA and morphological characters

Shovelnose sturgeon (Scaphirhynchus platorhynchus) caviar fisheries exist in several states throughout the Mississippi River drainage. Management of these fisheries may benefit from information about genetic stock structure. Sixteen microsatellite loci and morphological analysis were used to examine geographic stock structure of shovelnose sturgeon among seven geographic locations: five within continuous shovelnose sturgeon habitat, and two isolated by artificial barriers. Tissue samples were collected from 1999 to 2006 from the upper Missouri, Platte, lower Missouri, middle Mississippi, Ohio, Wabash, and Atchafalaya rivers. Geographic samples of shovelnose sturgeon samples could be separated into three groups with discriminate function analysis of four morphological characters. The microsatellite loci were highly variable (allelic richness range 5.65–13, observed heterozygosity range 0.64–0.89). Bayesian clustering did not identify multiple groups in the genetic data. However, significant genetic differentiation (θ_ST = 0.017, P < 0.0001) was observed among a priori defined geographic samples and all pairwise estimates of θ_ST were significant. Assignment testing among a priori defined groups indicated that the sturgeon from the upper Missouri, Platte, and Atchafalaya rivers had the highest assignment scores and thus were most distinct, while the lower Missouri and the middle Mississippi were less distinct and a larger fraction of the sturgeon from these rivers was genetically assigned to other rivers. The Ohio and Wabash rivers were genetically most similar. A Mantel test revealed a positive relationship between genetic and geographic distance (r = 0.464, P = 0.055) that was not statistically significant. The level of genetic differentiation observed at both molecular and morphological characters suggests that multiple shovelnose sturgeon populations may exist within the studied area, yet demographic factors and possible gene flow may have minimized the amount of genetic differentiation among locations.     

Phenological adaptations of a colonizing insect: The southwestern corn borer,Diatraea grandiosella

Summary The developmental rate, critical photoperiod, and diapause intensity were determined for three populations of the southwestern corn borer, Diatraea grandiosella, from Missouri, Mississippi and Kansas. Mississippi larvae grew at the highest rate and Missouri larvae grew at the lowest rate. The zero developmental temperatures (°C) for the Missouri population were estimated from regression lines as follows: 10.5° (eggs), 10.8° (diapausing larvae), 13.3° (non-diapausing larvae) and 11.4° (pupae). The required heat units were: 85° (eggs), 588° (diapausing larvae), 333° (non-diapausing larvae) and 149° days (pupae). However, the observed low temperature limit for larval growth under constant temperature regimes was approximately 17°C.The critical day lengths for diapause induction observed at 25°C were: 15 h 11 min (Missouri); 15 h 20 min (Mississippi); and 15 h 22 min (Kansas). The photoperiodic response of the Mississippi larvae was more or less retained at 30°C, whereas the response of the Missouri larvae was completely suppressed at this temperature. Diapause was most easily terminated in the Kansas larvae. The most intense diapause was observed in the Mississippi larvae.Model seasonal life cycles of the three geographic populations were constructed using photothermograms. Although the models showed good agreement with the field situation for the Missouri and the Kansas populations, some unknown factor(s) remains to account for an extremely long critical photoperiod in the Mississippi population.Contribution from the Missouri Agricultural Experiment Station, as journal series no. 9001     

Sediment microbial enzyme activity as an indicator of nutrient limitation in the great rivers of the Upper Mississippi River basin

We compared extracellular enzyme activity (EEA) of microbial assemblages in river sediments at 447 sites along the Upper Mississippi, Missouri, and Ohio Rivers with sediment and water chemistry, atmospheric deposition of nitrogen and sulfate, and catchment land uses. The sites represented five unique river reaches—impounded and unimpounded reaches of the Upper Mississippi River, the upper and lower reaches of the Missouri River, and the entire Ohio River. Land use and river chemistry varied significantly between rivers and reaches. There was more agriculture in the two Upper Mississippi River reaches, and this was reflected in higher nutrient concentrations at sites in these reaches. EEA was highest in the two Upper Mississippi River reaches, followed by the lower Missouri River reach. EEA was generally lowest in the upper Missouri River reach. Canonical correlation analysis revealed a strong correlation between EEA and the suite of water and sediment chemistry variables, and the percent of the catchment in anthropogenically dominated land uses, including agriculture and urban development. Nutrient ratios of the waters and sediments suggested carbon (C), nitrogen (N), or phosphorus (P) limitation at a large number of sites in each reach. C-limitation was most pronounced in the unimpounded Mississippi River and lower Missouri River reaches; N-limitation was prevalent in the two Missouri River reaches; and P-limitation dominated the Ohio River. Linking microbial enzyme activities to regional-scale anthropogenic stressors in these large river ecosystems suggests that microbial enzyme regulation of carbon and nutrient dynamics may be sensitive indicators of anthropogenic nutrient and carbon loading.     

Flooded flatheads: evidence of increased growth in Mississippi River <Emphasis Type="Italic">Pylodictis olivaris</Emphasis> (Pisces: Ictaluridae) following the Great Midwest Flood of 1993

Both the Mississippi and Missouri rivers experienced major flooding in the spring and summer of 1993, inundating much of their floodplains for long periods, and allowing fish access to vast but previously inaccessible benthic foraging areas. In response to the latter, we hypothesized that flathead catfish (Pylodictis olivaris) would exhibit altered post-flooding growth as compared to like-aged fish whose lives were flood-free. To test this hypothesis, we obtained the left pectoral fin spines from hoop-netted Mississippi River flathead catfish collected in the vicinity of Crystal City, Missouri, USA, and conducted an individual-based back-calculated growth comparison of individuals captured pre-flood (1991) and post-flood (1996). For like-aged cohorts, our back-calculated length-at-age data clearly show that the post-flooding growth in 1996-collected fish was superior to that of the 1991-collected fish that had experienced less exceptional flow regimes. This outcome suggests that increased forage or/and foraging opportunities can be a consequence of flooding, and that flathead catfish are able to capitalize on this. The natural flooding regime of big rivers may thus be an important contributor to the growth and success of certain fish species; consequently, modifying the river’s natural hydrograph and channel morphology may be detrimental. Handling editor: J. A. Cambray An erratum to this article is available at .     

Conforming to the status flow: The influence of altered habitat on fish body-shape characteristics

1. Anthropogenic stream impoundments often result in altered flow regimes and reservoir formations. Functional fish characteristics may change in response to altered flow conditions, and understanding the underlying mechanisms at work (e.g. natural selection and phenotypic plasticity) is of great importance in both evolutionary and conservation contexts. Cyprinella lutrensis is a widely distributed minnow native to stream systems in Missouri, U.S.A. Following impoundment of the Osage River, C. lutrensis currently occurs in the resultant Truman Reservoir and surrounding tributaries. To examine the impacts of this habitat alteration on functional fish characteristics, we hypothesised that C. lutrensis populations in altered systems with no-flow would show an overall less streamlined body shape as compared to those in flowing systems, assuming body streamlining is beneficial for locomotion in flowing environments.
2. We conducted field and laboratory studies to test effects of flow and no-flow on C. lutrensis body shape. Geometric morphometric techniques were used to quantify body shape in field and laboratory individuals, and non-structural lipid extractions were conducted to assess whether fat storage contributes to body shape under flow and no-flow treatments.
3. Morphological comparisons between C. lutrensis from Truman Reservoir and surrounding streams yielded significant differences in body shape, with greater body streamlining in stream populations. A laboratory experiment using artificial stream units revealed similar significant differences in body streamlining between flow and no-flow treatments within a single generation. In addition, individuals under no-flow treatment had significantly higher levels of non-structural lipids compared to flow treatment individuals.
4. Our results indicate that changes in flow regime can alter the evolutionary trajectories (i.e. related to morphology) of fish populations. We suggest that body shape alterations are likely to occur within and across generations in a cumulative manner, therefore, it is plausible that both natural selection and plastic response mechanisms play roles in defining minnow body shape following flow alteration.

     

Use of genetic tags to identify captive-bred pallid sturgeon (<Emphasis Type="Italic">Scaphirhynchus albus</Emphasis>) in the wild: improving abundance estimates for an endangered species

The ability to distinguish captive-bred and natural-origin individuals in the wild is critical for evaluating the impact of captive breeding programs on natural populations. Continued persistence of endangered pallid sturgeon (Scaphirhynchus albus) in the Missouri River is largely dependent on captive breeding efforts that spawn natural-origin adults in fish hatcheries and release their progeny into the wild. Prior to release, hatchery-origin individuals are physically marked so they can be distinguished from natural-origin individuals when recaptured. During the years 2004–2006, 24 unmarked juvenile pallid sturgeon tissue samples were collected in the Missouri River downstream of Gavins Point Dam, South Dakota, USA that were presumed natural-origin. However, these individuals were similar in size to hatchery-origin fish released in this area raising concerns that these individuals were actually hatchery-origin fish with lost or malfunctioning tags. We used microsatellite based parentage analysis to determine if the unmarked fish were members of hatchery families that had been released in this area. This retrospective genetic tagging approach revealed that 23 of 24 unmarked fish were indeed hatchery-origin. The origin of the remaining individual remains unknown because genetic samples were not available from all of the families released below the dam and the unassigned fish may have originated from one of these un-sampled families. These results provide important insight into the conservation status of endangered pallid sturgeon as well as provide data important for guiding management decisions. Our results also demonstrate the efficacy of using genetic tags as an alternative or complimentary approach to physically marking individuals.     

Microsatellite variation among River Sturgeons of the genus Scaphirhynchus (Actinopterygii: Acipenseridae): a preliminary assessment of hybridization*

Microsatellite variation from 13 disomic loci is reported for a total of 208 individuals of the genus Scaphirhynchus. This includes 105 individuals of the pallid sturgeon (Scaphirhynchus albus) from the lower Mississippi River, 11 pallid sturgeon from the Upper Mississippi and Missouri rivers, 65 shovelnose sturgeon (S. platorynchus) from the lower Mississippi River, six Alabama sturgeon (S. suttkusi), and 21 individuals of sturgeon identified as intermediate between S. albus and S. platorynchus. Results indicate that all five of the above population/species units are significantly differentiated from one another based on pairwise F_ST estimates. Locus Spl‐7 was diagnostic for the Alabama sturgeon and serves to further differentiate this allopatric species from other Scaphirhynchus. Classification of genotypes with and without a priori designations failed to clearly delineate the species and intermediates in the latter case but was successful for the species but not in the intermediates in the former case. The presence of six unique alleles in five of the 21 morphologically ‘intermediate’ sturgeon examined requires additional evaluation but suggests that these individuals are possibly not the result of hybridization. We hope that raising these important issues will bring all stakeholders to the table to establish a concerted effort needed for both morphological and molecular analyses to adequately address the question of hybridization and the origin of the morphological variation in these fishes.     

Stock structure of pallid sturgeon analyzed with microsatellite loci

Recovery efforts for the endangered pallid sturgeon (Scaphirhynchus albus) include supplementation of wild stocks with hatchery reared progeny. Identifying the extent of genetic stock structure, which has previously been detected in samples from the range extremes, will help to determine whether stock transfers might be harmful. DNA microsatellite genotypes were screened in pallid sturgeon from the upper Missouri River, lower Missouri River, middle Mississippi River and Atchafalaya River and analyzed using a combination of Bayesian model‐based and more traditional F‐statistic based methods to characterize genetic differentiation. Scaphirhynchus specimens were collected by researchers active in the recovery effort and genotypes were screened at 16 microsatellite loci. Because there is considerable genetic and morphological overlap between pallid sturgeon, shovelnose sturgeon, and their hybrids, a combination of morphological and genetic techniques were used to eliminate shovelnose and possible hybrids from the sample. Genetic differentiation was detected among samples (overall θ = 0.050, P = 0.001). Pairwise θ, genetic distances, and Bayesian assignment testing reveal that pallid sturgeon from the upper Missouri River are the most distinct group with pairwise comparisons of pallid sturgeon among all the remaining samples exhibiting lower θ values, higher genetic distances, and self assignment scores. Our results indicate that using local broodstock, when available, should be used for pallid sturgeon propagation. If local broodstock are not available, geographically proximate individuals would limit genetic differences between native and stocked individuals.     

Trout (<Emphasis Type="Italic">Salmo trutta</Emphasis>) mitochondrial DNA polymorphism in the centre of the marble trout distribution area

The marble trout, a lineage of the Salmo trutta complex, is endemic to the Southern Alpine region. Although it is endangered throughout its entire distribution range, population genetic data were lacking for the central area, including the upper Etsch/Adige River system (South Tyrol, Northern Italy). A total of 672 Salmo trutta specimens, comprising phenotypic marble trout and phenotypic brown trout, from 20 sampling sites throughout South Tyrol were analysed by sequencing the complete mitochondrial DNA control region. Thirteen distinct haplotypes were identified, which clustered within three major genetic lineages: the Marmoratus (MA), the Atlantic (AT) and the Danubian (DA) lineage. 41.7% of the investigated individuals carried haplotypes of the MA lineage, 47.9% of the AT lineage and 10.4% of the DA lineage. It is noticeable that AT haplotypes were present at all sampling sites and no “pure” marble trout population with exclusively MA haplotypes was found. This points to a considerable impact of stocking with allochthonous brown trout, given that there is no evidence for natural colonisation by individuals of the AT lineage. However, our data indicate, for at least four localities, a limited gene flow between the native marble trout and hatchery-reared strains. Future conservation and rehabilitation measures will thus have to concentrate on the identification of remnant pure marble trout individuals from such mixed populations. Handling editor: C. Sturmbauer     

Phylogeographic Analyses Suggest Multiple Lineages of Crystallaria asprella (Percidae: Etheostominae)

The crystal darter, Crystallaria asprella, exists in geographically isolated populations that may be glacial relicts from its former, wide distribution in the Eastern U.S. An initial phylogeographic survey of C. asprella based upon the mitochondrial cytochrome b (cyt b) gene indicated that there were at least four distinct populations within the species: Ohio River basin, Upper Mississippi River, Gulf coast, and lower Mississippi River. In particular, the most divergent population was the most recently discovered, from the Elk River, WV, in the Ohio River basin, and it was postulated that this population represents an undescribed, potentially threatened species. However, differentiation observed at a single gene region is generally not considered sufficient evidence to establish taxonomic status. In the present study, nucleotide variation at the mitochondrial control region and a nuclear S7 ribosomal gene intron were compared to provide independent verification of phylogeographic results between individuals collected from the same five disjunct populations previously surveyed. Variation between populations at the control region was substantial (except between Gulf drainages) and was concordant with patterns of sequence divergence from cyt b. Only the Elk River population was resolved as monophyletic based upon nuclear S7, but significant differences based upon Φ_ST statistics were observed between most populations. Morphometric data were consistent with molecular data regarding the distinctiveness of the Elk River population. It is proposed that populations of C. asprella consist of at least four distinct population segments, and that the Elk River group likely constitutes a distinct species.