Isolation and characterization of microsatellites in Chinese sturgeon,Acipenser sinensis

From (GATA)_n and (AAAG)_n enriched genomic libraries for the Chinese sturgeon (Acipenser sinensis), 50 primer pairs were developed using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Forty‐six primer pairs exhibited highly polymorphic with two to 11 alleles per locus, while the rest four displayed monomorphic. These markers yielded 246 alleles in a survey of eight specimens of wild A. sinensis. Average observed heterozygosity ranged from 0.13 to 1.00. These loci should provide sufficient levels of genetic diversity to allow parentage analysis for artificial stocking management and delineation of fine‐scale population structure.     

Development of new microsatellite primers for green and white sturgeon

Sixty-eight primer sets for microsatellite loci were developed from microsatellite motif enriched genomic libraries of pooled DNA from the polyploid green and white sturgeon (Acipenser medirostris and A. transmontanus). Four individuals from each species were screened for polymorphism at these loci. Forty-eight loci amplified in both species, and some exhibited species-specific amplification for white or green sturgeon (8 and 12 loci, respectively). The number of alleles per locus ranged from one to 12. At least 68% of the green and 65% of the white sturgeon loci we developed are polysomic.     

The use of microsatellite loci for identification of sturgeon species (Acipenseridae) and hybrid forms

The genetic polymorphism of ten sturgeon species that inhabit the territory of the Russian Federation (Russian sturgeon, Siberian sturgeon, Amur sturgeon, Sakhalin sturgeon, Persian sturgeon, ship sturgeon, sterlet, stellate sturgeon, beluga, and kaluga) was examined at five microsatellite loci (Afug41, Afug51, An20, AoxD161, AoxD165) (in total, 3821 individuals). The examined loci were successfully amplified with the same primer set in all species tested and demonstrated a high level of variation. Alleles specific to different species have been identified, which allows them to be used to identify species of sturgeon and their food products. In addition, the possibility of identifying hybrid forms was demonstrated. The assignment test performed in the STRUCTURE software program showed a high probability of correctly assigning each individual to its species based on genotyping with five microsatellite loci examined (96–98%, on average). However, for Russian and Persian sturgeon, the rate of proper species assignment was considerably lower (75 and 84%, respectively).

     

Comparative stable isotope analyses of green sturgeon (Acipenser medirostris) and white sturgeon (A. transmontanus) in the San Francisco estuary

Green sturgeon (Acipenser medirostris) and white sturgeon (A. transmontanus) are closely related, sympatric species that inhabit the San Francisco estuary. Green sturgeon have a more marine life history but both species spawn in the Sacramento River and reside for some duration in San Francisco Bay. These sturgeons are of conservation concern, yet little is known about their dietary competition when they overlap in space and time. To examine evidence of dietary differentiation, we collected whole blood and blood plasma from 26 green sturgeon and 35 white sturgeon in San Francisco Bay. Using carbon and nitrogen stable isotope analyses, we compared their relative trophic levels and foraging locations along the freshwater to marine gradient. Sampling blood plasma and whole blood allowed comparison of dietary integration over shorter and longer time scales, respectively. Plasma and whole blood δ¹³C values confirmed green sturgeon had more marine dietary sources than white sturgeon. Plasma δ¹⁵N values revealed white sturgeon fed at lower trophic levels than green sturgeon recently, however, whole blood δ¹⁵N values demonstrated the two species fed at the same trophic level over longer time scales. Larger individuals of both species had higher δ¹³C values than smaller individuals, reflecting more marine food sources in adulthood. Length did not affect δ¹⁵N values of either species. Isotope analyses supported the more marine life history of green than white sturgeon and potentially highlight a temporary trophic differentiation of diet between species during and preceding the overlapping life stage in San Francisco Bay.     

Isolation of microsatellite loci from the endemic and endangered Adriatic sturgeon (<Emphasis Type="Italic">Acipenser naccarii</Emphasis>)

With the aim of elaborating a breeding plan on a captive stock of the highly endangered Adriatic sturgeon (Acipenser naccarii), a total of 10 polymorphic microsatellite loci were isolated from an enriched library. The results of cross amplification of additional 8 loci previously isolated from A. oxyrinchus, A. fulvescens and Scaphyrinchus platorhynchus are also reported. Given the tetraploid condition of the species the genetic variability was estimated basing on the number of alleles per individuals and the average band sharing.     

Development and standardization of disomic microsatellite markers for lake sturgeon genetic studies

Lake sturgeon (Acipenser fulvescens) are of conservation concern in North America. To facilitate the recovery of this fish species, an understanding of their population genetic structure is necessary to develop and implement spatially and temporally appropriate management actions. Until recently, few genetic data using nuclear loci have been collected, primarily due to the paucity of suitable genetic markers because most microsatellite loci in lake sturgeon appeared to be tetrasomic. The authors identified nine microsatellite loci (from 254 examined) that were putative polymorphic disomic loci and tested their conformance to a disomic mode of inheritance using three lake sturgeon families. The objectives of the study were to: (i) confirm the disomic status of the nine loci through inheritance testing, and (ii) standardize the genetic markers among participating laboratories. At all nine loci, disomic inheritance were confirmed, and all nine loci segregated independently in the 26 of 36 loci pairs possible to test. One of the nine loci showed non‐Mendelian segregation, possibly due to meiotic drive and/or selection. Three progeny had peak patterns inconsistent with disomy at one or more loci. The nine loci when combined with four microsatellite loci previously confirmed in other studies as disomic in lake sturgeon now yield a suite of 13 microsatellite markers. These 13 markers have been standardized among four other laboratories to facilitate building an inter‐laboratory genetic database for lake sturgeon.     

Behavioural salinity preferences of juvenile green sturgeon Acipenser medirostris acclimated to fresh water and full‐strength salt water

To quantify the salinity preference of juvenile green sturgeon Acipenser medirostris, two groups of A. medirostris [140 days post hatch (dph); total length (L_T) 38·0–52·5 cm] were acclimated to either near fresh water (mean ± s.e . salinity = 3·2 ± 0·6) or full‐strength salt water (34·1 ± 1·2) over 8 weeks. Following acclimation, the two groups were divided into experimental and control groups, where experimental A. medirostris from both freshwater and saltwater acclimations were individually introduced (200–220 dph) into a rectangular salinity‐preference flume (maximum salinity gradient: 5–33). Control A. medirostris were presented with only their acclimation water (fresh water or salt water) on both sides of the flume. It was demonstrated that A. medirostris acclimated to both salt water and fresh water spent a significantly greater amount of time on the side of the testing area with the highest salinity concentration (P < 0·05 and P < 0·001, respectively) while control A. medirostris spent an equal amount of time on each side of the flume. These findings indicate that juvenile A. medirostris are not only capable of detecting salt water within the first year of their lives but perhaps are actively seeking out saline environments as they move through a watershed. Establishing A. medirostris salinity preferences provides a better understanding of the early life history of this threatened species, shedding light on possible outmigration timing.     

Evaluating the effects of controlled flows on historical spawning site access,reproduction and recruitment of lake sturgeon Acipenser fulvescens

Lake sturgeon Acipenser fulvescens spawn at the base of Kakabeka Falls, a 39 m waterfall on the Kaministiquia River, a tributary to Lake Superior. Access to this historical spawning site can be restricted or delayed due to hydroelectric flow fluctuations that coincide with the A. fulvescens spawning season. The objectives of this study were to determine (a) the necessary flow conditions that facilitate spawning site access; (b) quantity and duration of flow required for successful spawning and dispersal of larvae; and (c) evaluate recruitment of juvenile A. fulvescens in relation to flow. A. fulvescens spawning migrations were tracked using a stationary telemetry receiver that logged the movements of 166 A. fulvescens fitted with radio-transmitters. Unrestricted access to the spawning site was facilitated when spawning flow was controlled at 23 m³ s⁻¹ in 2004 and 17 m³ s⁻¹ in 2006. Fluctuating (0.5–8.5 m³ s⁻¹) and delayed spawning flows resulted in restricted and delayed access to the spawning site. Flow duration for successful egg incubation, hatch and larval dispersal was determined by sampling larvae using drift nets and quantified using cumulative temperature units (CTU). Over 10 years, 10,083 larvae were captured between 31 May and 20 July with 97% of the drift occurring prior to 30 June. From the date of first spawning to the end of larval dispersal took an average of 38.6 days, and the mean CTU value was 398.6. In general, a minimum flow of approximately 14.5 m³ s⁻¹ from the date of initial spawning to the accumulation of c. 400 CTU ensured successful hatch and larval dispersal. During the timeframe of this study, recruitment was variable. This study described the complex and variable reproductive life history of A. fulvescens and defined spawning flow requirements ecologically, which can be used to develop operational provisions at hydropower facilities to ensure successful reproduction.     

Genetic effects of habitat restoration in the Laurentian Great Lakes: an assessment of lake sturgeon origin and genetic diversity

Lake sturgeon (Acipenser fulvescens) have experienced significant habitat loss, resulting in reduced population sizes. Three artificial reefs were built in the Huron‐Erie corridor in the Great Lakes to replace lost spawning habitat. Genetic data were collected to determine the source and numbers of adult lake sturgeon spawning on the reefs and to determine if the founder effect resulted in reduced genetic diversity. DNA was extracted from larval tail clips and 12 microsatellite loci were amplified. Larval genotypes were then compared to 22 previously studied spawning lake sturgeon populations in the Great Lakes to determine the source of the parental population. The effective number of breeders (N_b) was calculated for each reef cohort. The larval genotypes were then compared to the source population to determine if there were any losses in genetic diversity that are indicative of the founder effect. The St. Clair and Detroit River adult populations were found to be the source parental population for the larvae collected on all three artificial reefs. There were large numbers of contributing adults relative to the number of sampled larvae. There was no significant difference between levels of genetic diversity in the source population and larval samples from the artificial reefs; however, there is some evidence for a genetic bottleneck in the reef populations likely due to the founder effect. Habitat restoration in the Huron‐Erie corridor is likely resulting in increased habitat for the large lake sturgeon population in the system and in maintenance of the population's genetic diversity.     

Summer diet of juvenile lake sturgeon reintroduced into the Genesee and St. Regis Rivers,New York USA

The restoration of threatened species involves understanding multiple aspects of the life history and ecology of the target species. One important consideration in the restoration of threatened species is feeding ecology. We examined the summer diet of reintroduced juvenile lake sturgeon (Acipenser fulvescens) in the Genesee (n = 119, ages 1 and 2) and the St. Regis rivers (n = 40, ages 1, 2, 5, 6, and 7) of New York State, USA, in the summer of 2005. The most common taxa consumed were Diptera (88% of prey biomass) in the Genesee River and Ephemeroptera (35% of prey biomass) and Diptera (29%) in the St. Regis River. Diptera: (Chironomidae) was the major prey taxon in both benthic communities. Selectivity analysis showed a positive selection of Diptera by Genesee River lake sturgeon and an avoidance of Diptera by St. Regis River lake sturgeon. The St. Regis River lake sturgeon showed avoidance of Coleoptera, but positive selection of Ephemeroptera, Trichoptera, and Oligochaeta. Results indicate that the reintroduced juvenile lake sturgeon are growing and successfully finding prey in these two rivers and appear to actively select specific prey types.     

Species and hybrid identification of sturgeon caviar: a new molecular approach to detect illegal trade

Overexploitation of wild populations due to the high economic value of caviar has driven sturgeons to near extinction. The high prices commanded by caviar on world markets have made it a magnet for illegal and fraudulent caviar trade, often involving low‐value farmed caviar being sold as top‐quality caviar. We present a new molecular approach for the identification of pure sturgeon species and hybrids that are among the most commercialized species in Europe and North America. Our test is based on the discovery of species‐specific single nucleotide polymorphisms (SNPs) in the ribosomal protein S7, supplemented with the Vimentin gene and the mitochondrial D‐loop. Test validations performed in 702 specimens of target and nontarget sturgeon species demonstrated a 100% identification success for Acipenser naccarii, A. fulvescens, A. stellatus, A. sinensis and A. transmontanus. In addition to species identification, our approach allows the identification of Bester and AL hybrids, two of the most economically important hybrids in the world, with 80% and 100% success, respectively. Moreover, the approach has the potential to identify many other existing sturgeon hybrids. The development of a standardized sturgeon identification tool will directly benefit trade law enforcement, providing the tools to monitor and regulate the legal trade of caviar and protect sturgeon stocks from illicit producers and traders, hence contributing to safeguarding this group of heavily threatened species.     

Ionic and osmotic regulation capabilities of juvenile Gulf of Mexico sturgeon, Acipenser oxyrinchusde sotoi

The salinity tolerance, and hydromineral regulation capabilities of three size groups (small 110–170 g; medium 230–290 g, large 460–700 g; n=48 for each group) of 13-month-old juvenile Gulf of Mexico sturgeon were investigated. Fish (n=6 for each salinity) were transferred directly from freshwater (FW) to a series of experimental salinity treatments (0, 5, 10, 15, 20, 25, 30, and 35 parts per thousand (ppt)). Fish were also acclimated in brackish water (20 ppt) for 2 weeks and transferred to a salinity of 34 ppt. In this condition juvenile Gulf of Mexico sturgeon adapted to saltwater (SW) and maintained their hydromineral balance. FW adapted sturgeon (n=6) had an average blood hemotocrit of 28.2±0.8%, plasma osmolality of 260.7±1.6 mOsm kg⁻¹ H₂O, and plasma ion concentrations of 135.7±1.2 mM l⁻¹ Na⁺, 106.9±1.9 mEq l⁻¹ Cl⁻, and 2.9±0.1 mM l⁻¹ K⁺. In SW adapted sturgeon (n=8) blood parameters averaged 26.9±0.7% for hematocrit, 294.2±2.3 mOsm kg⁻¹ H₂O for osmolality, 152.0±1.7 mM l⁻¹ Na⁺, 149.2±1.4 mEq l⁻¹ for Cl⁻, and 3.1±0.1 mM l⁻¹ K⁺. The method of transfer (abrupt or slow acclimation) directly affected fish survival and the time they took to achieve ionic and osmotic regulation. This SW adaptation appears to be related to body size, the larger the fish the easier the adaptation process. A threshold size of about 170 g was apparent for the fish to adapt to saltwater after 2 weeks of acclimation. Chloride cells were present in both FW and SW adapted sturgeon with SW and brackish water fish having chloride cells significantly (P<0.05) more numerous (561±53 and 598±45 cells mm⁻²) and larger in size (41.0±3.85 and 34.2±4.49 μm²) than FW adapted sturgeon (10±1.0 cells mm⁻² and 22±2.53 μm²). Few chloride cells were observed in the opercular membrane, however, none were found in the pseudobranch and spiracle.     

Mitochondrial DNA variation, effective female population size and population history of the endangered Chinese sturgeon, Acipenser sinensis

The anadromous Chinese sturgeon (Acipenser sinensis), mainly endemic to the Yangtze River in China, is an endangered fish species. The natural population has declined since the Gezhouba Dam blocked its migratory route to the spawning grounds in 1981. In the near future, the completion of the Three Gorges Dam, the world's largest hydroelectric project, may further impact this species by altering the water flow of the Yangtze River. Little is currently known about the population genetic structure of the Chinese sturgeon. In this study, DNA sequence data were determined from the control region (D-loop) of the mitochondrial genome of adult sturgeons (n = 106) that were collected between 1995–2000. The molecular data were used to investigate genetic variation, effective female population size and population history of the Chinese sturgeon in the Yangtze River. Our results indicate that the reduction in abundance did not change genetic variation of the Chinese sturgeon, and that the population underwent an expansion in the past. AMOVA analysis indicated that 98.7% of the genetic variability occurred within each year's spawning populations, the year of collection had little influence on the diversity of annual temporary samples. The relative large effective female population size (N _ef) indicates that good potential exists for the recovery of this species in the future. Strikingly, the ratio of N _ef to the census female population size (N _f) is unusually high (0.77–0.93). This may be the result of a current bottleneck in the population of the Chinese sturgeon that is likely caused by human intervention. This revised version was published online in July 2006 with corrections to the Cover Date.     

Population assessment and movement patterns of lake sturgeon (Acipenser fulvescens) in the Manistee River, Michigan, USA

Lake sturgeon (Acipenser fulvescens) population characteristics and migration patterns were evaluated in the Manistee River system, Michigan. Sturgeon (n = 102) were captured using gillnets, tagged and released in the system between 2001 and 2005. Biological data (weight, length, age, sex and spawning status) were collected to calculate length–weight relationships and relative condition factor (K_n), determine age structure, and estimate the annual spawning population size. Sturgeon were also implanted with radio and sonic transmitters, and migration patterns of 22 sturgeon were determined. The weight–length equation for sturgeon in the Manistee River system was log₁₀ W = 3.17log₁₀TL?5.52 and did not differ by sex. K_n equaled 0.72 and did not differ by sex or spawning status. Ages ranged from age 5 to age 54. Sexually mature male and female sturgeon were captured over the course of the study, and the size of the spawning population ranged from 21 to 66 spawners. Sturgeon migration movements up and down the Manistee River varied with date, water temperature, and discharge. The sturgeon appeared to use two different spawning sites in the Manistee River. The Manistee River system supports a remnant lake sturgeon population; the presence of a wide range of age classes and availability of a limited number of suitable spawning sites suggest continued recruitment to the population.     

Isolation and characterization of microsatellites in the Adriatic sturgeon (Acipenser naccarii)

We isolated and characterized seven polymorphic microsatellite loci from a partial genomic library of the Adriatic sturgeon (Acipenser naccarii) enriched for GATA repeats. Variability was tested in 20 specimens of this endangered species. Our data support the tetraploid condition for A. naccarii and will be useful for the investigation of the remnant genetic variability of this species as well as for genetic tagging studies.     

Experimental assessment of predation risk for juvenile green sturgeon,Acipenser medirostris,by two predatory fishes

Predation is a common cause of early life stage mortality in fishes, with reduced risk as individuals grow and become too large to be consumed by gape-limited predatory fishes. Large-bodied species, such as sturgeon, may reach this size-refuge within the first year. However, there is limited understanding of what this size threshold is despite the value of this information for conservation management. We conducted laboratory-based predation experiments on juvenile green sturgeon, Acipenser medirostris, to estimate vulnerability to predation during outmigration from their natal reaches in California to the Pacific Ocean. Two highly abundant and non-native predatory fish species (largemouth bass, Micropterus salmoides, and striped bass, Morone saxatilis) were captured in the wild to be tested with developing juvenile green sturgeon from the UC Davis Green Sturgeon Broodstock Program. Experimental tanks, each containing five predators, received thirty prey for 24-hr exposures. Between sturgeon prey trials, predators were exposed to alternative prey species to confirm predators were exhibiting normal feeding behaviors. In addition to green sturgeon mortality data, trials were video recorded and predatory behaviors were quantified. Overall, these predator species displayed much lower rates of predation on juvenile green sturgeon than alternate prey. Predation decreased with green sturgeon size, and predation risk diminished to zero once sturgeon reached a length threshold of roughly 20–22 cm total length, or between 38% and 58% of predator total length. Behavioral analyses showed low motivation to feed on green sturgeon, with both predators attempting predation less frequently as sturgeon grew. Results of this study imply that optimizing growth rates for larval and juvenile sturgeon would shorten the time in which they are vulnerable to predation. Future experiments should assess predation risk of juvenile green sturgeon by additional predator species common to the Sacramento-San Joaquin watershed.     

Phylogenetic relationships of Amur sturgeon Acipenser schrenckii Brandt, 1869 based on 18S rDNA sequencing

The phylogenetic relationships of Amur sturgeon A. schrenckii Brandt, 1869 with related species have been analyzed based on sequencing of the 18S rDNA small subunit. The complete sequence (1746 bp) of 18S rDNA has been estimated in seven individual A. schrenckii clones. The results show that the rDNA mutation profile of A. schrenckii 18S is very similar to that of A. fulvescens (Genbank data). Structural-functional and phylogenetic analyses allowed us to identify a presumably expressed variant, as well as taxon-specific mutation (adenine insertion after position 658 bp), for A. schrenckii 18S rDNA. Phylogenetic reconstructions performed with various approaches (NJ, MP, ML and Bayesian) support the monophyly of the genus Acipenser and point (1), based on which, in accordance with the classification based on ecological and morphological data (Artyukhin, 2006), the Amur sturgeon is closer to the sterlet than the Baltic sturgeon and (2) to substantial differentiation between North American (A. fulvescens) and Eurasian (A. schrenckii, A. ruthenus, and A. sturio) species of Acipenseridae.     

Evidence of spawning by green sturgeon, Acipenser medirostris, in the upper Sacramento River, California

This study reports the only direct evidence of spawning of green sturgeon, Acipenser medirostris, in the upper Sacramento River, CA. Two green sturgeon eggs were collected with substrate mats immediately below Red Bluff Diversion Dam. One green sturgeon larva was collected with a larval net at Bend Bridge. We concluded that green sturgeon spawn in the upper Sacramento River, both above and below RBDD. Temperature ranges in the study area (10–15°C) are similar to conditions used in successful artificial rearing of green sturgeon and do not appear to be a limiting factor to successful spawning of green sturgeon; however, suitable habitat upstream of RBDD is inaccessible when dam gates are lowered.     

Use of Washington Estuaries by Subadult and Adult Green Sturgeon

Green sturgeon, Acipenser medirostris, are the most marine-oriented of North American sturgeons. However, their estuarine/marine distribution and the seasonality of estuarine use are largely unknown. We used acoustic telemetry to document the timing of green sturgeon use of Washington estuaries. In the summers of 2003 and 2004, uniquely coded acoustic transmitters were surgically implanted in green sturgeon captured using commercial gillnets. All sturgeon tagged were greater than 1.2 m total length. They were caught, tagged, and released in both Willapa Bay (n = 49) and Columbia River (n = 11) estuaries. We deployed an array of four fixed- site acoustic receivers in Willapa Bay to detect the estuarine entry and exit of these and any of over 100 additional green sturgeon tagged in other systems during 2003 and 2004. Green sturgeon occurred in Willapa Bay in summer when estuarine water temperatures exceeded coastal water temperatures by at least 2°C. They exhibited rapid and extensive intra- and inter- estuary movements and green sturgeon from all known spawning populations were detected in Willapa Bay. We hypothesize that green sturgeon optimize their growth potential in summer by foraging in the relatively warm, saline waters of Willapa Bay and we caution that altering the quality of estuarine habitats could negatively affect this species throughout its range.     

Interannual variation in effective number of breeders and estimation of effective population size in long‐lived iteroparous lake sturgeon (Acipenser fulvescens)

Quantifying interannual variation in effective adult breeding number (N_b) and relationships between N_b, effective population size (N_e), adult census size (N) and population demographic characteristics are important to predict genetic changes in populations of conservation concern. Such relationships are rarely available for long‐lived iteroparous species like lake sturgeon (Acipenser fulvescens). We estimated annual N_b and generational N_e using genotypes from 12 microsatellite loci for lake sturgeon adults (n = 796) captured during ten spawning seasons and offspring (n = 3925) collected during larval dispersal in a closed population over 8 years. Inbreeding and variance N_b estimated using mean and variance in individual reproductive success derived from genetically identified parentage and using linkage disequilibrium (LD) were similar within and among years (interannual range of N_b across estimators: 41–205). Variance in reproductive success and unequal sex ratios reduced N_b relative to N on average 36.8% and 16.3%, respectively. Interannual variation in N_b/N ratios (0.27–0.86) resulted from stable N and low standardized variance in reproductive success due to high proportions of adults breeding and the species' polygamous mating system, despite a 40‐fold difference in annual larval production across years (437–16 417). Results indicated environmental conditions and features of the species' reproductive ecology interact to affect demographic parameters and N_b/N. Estimates of N_e based on three single‐sample estimators, including LD, approximate Bayesian computation and sibship assignment, were similar to annual estimates of N_b. Findings have important implications concerning applications of genetic monitoring in conservation planning for lake sturgeon and other species with similar life histories and mating systems.