The Holocene occurrence of Acipenser spp. in the southern North Sea: the archaeological record

Archaeological sturgeon remains from the southern North Sea basin used to be automatically attributed to Acipenser sturio, since this was the only acipenserid species believed to occur there. These species identifications, however, were in need of revision after a growing number of indications were found for the historical presence of Acipenser oxyrinchus in western Europe. In this study, morphological and genetic data on sturgeon remains from archaeological sites along the southern North Sea are revised. A large number of Dutch, Belgian, British and some French archaeological sturgeon remains, dating from the Mesolithic up to Late Modern times, are morphologically examined and fish sizes are reconstructed. This study of >7000 acipenserid bones proves the sympatric occurrence of European sturgeon A. sturio and Atlantic sturgeon A. oxyrinchus in the southern North Sea at least since the Neolithic (fourth millennium BC onwards), with A. oxyrinchus remains always outnumbering those of A. sturio. Human influence is documented by the decrease in finds through time, but no clear evidence was found for a diachronic change in fish lengths that could possibly be related to fishing pressure.     

First archaeozoological identification of Atlantic sturgeon (Acipenser oxyrinchus Mitchill 1815) in France

To this day, the only sturgeon to be listed on the French vertebrate inventory is the European sturgeon (Acipenser sturio Linnaeus, 1758). The recent study of sturgeon remains on various French archaeological sites shows the presence of another species: the Atlantic sturgeon (A. oxyrinchus Mitchill 1815). This species already existed in the French Atlantic region at the end of the Neolithic Age 5000 years ago and was still to be found 3000 years later. Thus the A. oxyrinchus determined in several Baltic medieval sites are neither the only nor the first sturgeons to have inhabited European waters. Sturgeon restoration projects in European rivers necessitate a precise determination of the native species. In the case of relict or extinct species, the bone remains found on archaeological sites represent the most reliable source of information. This discovery will also be the starting point of palaeogenetical research (mitochondrial and cellular aDNA) and will give information about the genetic diversity of these threatened or recently extinct populations.     

Comparison of karyotypes of <Emphasis Type="Italic">Acipenser oxyrinchus</Emphasis> and <Emphasis Type="Italic">A. sturio</Emphasis> by chromosome banding and fluorescent in situ hybridization

A highly debated problem in Acipenseridae taxonomy is whether Acipenser oxyrinchus (North American Atlantic sturgeon) and A. sturio (European Atlantic sturgeon) are true species: a detailed comparison of their karyotypes could provide relevant information. Here we describe for the first time the karyotype of A. oxyrinchus (2n = 121 ± 3), and its features, among which the constitutive heterochromatin, revealed by C-banding technique, the distribution of telomeric regions, and the 5S rRNA genes, detected by FISH. The results reveal that A. oxyrinchus and A. sturio karyotypes and features are quite similar. Moreover, comparing the results obtained through hybridization by FISH with HindIII and PstI satellite DNA in these and in other sturgeon species, no hybridization signals are detected in A. sturio and A. oxyrinchus, while A. stellatus and A. gueldenstaedtii show hybridization. Thus A. sturio and A. oxyrinchus appear very similar from a cytogenetic point of view: these and molecular data repeatedly point out that A. sturio and A. oxyrinchus represent a sister clade in comparison to all other sturgeon species up to now studied.     

Complete mitochondrial genome sequence and phylogenetic position of the Amu Darya sturgeon,Pseudoscaphirhynchus kaufmanni (Acipenseriformes: Acipenseridae)

In this study, we successfully assembled the complete mitochondrial genome of the Amu Darya sturgeon Pseudoscaphirhynchus kaufmanni. Based on this mitochondrial genome and previously published mitochondrial genomes of members of the Acipenseridae family, we assessed the phylogenetic position of P. kaufmanni using maximum likelihood and Bayesian inference for phylogeny reconstruction. The resultant phylogenetic trees were well-resolved, with congruence between different phylogenetic methods. This robust phylogenetic analysis elucidated the relationship among the four acipenserid genera and strongly supported the division of the family into three main clades. Evaluation of molecular phylogeny using maximum likelihood and Bayesian analysis led to the following conclusions: (a) the most basal position within the Acipenseridae remains in the clade containing Acipenser oxyrinchus and Acipenser sturio; (b) the genus Scaphirhynchus belongs to the Atlantic clade and is a sister group of the remaining species of the clade; and (c) the close relationship between P. kaufmanni and Acipenser stellatus is well supported.     

Historical presence of the sturgeon Acipenser sturio in the Rhône basin determined by the analysis of ancient DNA cytochrome b sequences

Sturgeon disappeared from the Rhône River in the mid 70’s without certitude about which species it was and about the existence of a sympatry between European sturgeon, Acipenser sturio, and Adriatic sturgeon, A. naccarii, in this watershed. In order to reach a reliable specific determination of this extinct sturgeon population, archaeozoological remains of the Jardin d’Hiver in Arles city, on the Rhône river banks, were genetically analysed, following strict criteria of authentication for the ancient DNA work. The rich collection of Arles sturgeon bone remains stems from human activities between the 6th and the 2nd Century BC. Sequences of 86 bp of the cytochrome b gene were obtained on four bones, from different anatomical parts of the fish and from different archaeological layers. All gave A. sturio diagnostic sequences. This preliminary analysis is an essential first step in the project of sturgeon reintroduction in the Rhône River. Thus, further analyses on a larger sample are necessary to comfort this result and to solve the question of sympatry with A. naccarii.     

Current Occurrence of the Atlantic Sturgeon Acipenser oxyrinchus in Northern Spain: A New Prospect for Sturgeon Conservation in Western Europe

Acipenser oxyrinchus is considered extirpated in Europe, but numerous breeding populations still exist on the Atlantic coast of North America. An adult female A. oxyrinchus, 2500 mm total length and 120 kg wet weight, was accidentally fished on 24 November 2010 near the coast of Gijón, Asturias, Spain. The fish was identified by its morphological pattern as well as by mitochondrial and nuclear DNA analyses. Because the sturgeon was found far away from any known breeding area, it was considered a stray or vagrant specimen. It certainly has a natural origin, but its eventual birthplace could not be determined. Because its current occurrence was unknown in southwestern Europe until now, the species is not cataloged or protected in this area. Therefore, the residual European stocks of A. oxyrinchus ought to be listed as Critically Endangered (CR) according to the IUCN categories. Likewise, it is imperative for southwestern European countries with an historic or recent occurrence of A. oxyrinchus to protect the species through domestic and international legislation. The present sympatric occurrence of A. sturio and A. oxyrinchus raises new challenges about key questions, such as the species selection for restoration program in European countries. Accurate monitoring is mandatory to obtain appropriate information for an assessment of the current occurrence of A. oxyrinchus in southwestern Europe.     

Assessing the potential of the European Atlantic sturgeon Acipenser sturio to control bivalve invasions in Europe

This pilot study explored the potential of juvenile European Atlantic sturgeon Acipenser sturio to feed on two invasive bivalve species, the Asian clam Corbicula fluminea and the Eurasian zebra mussel Dreissena polymorpha. Preliminary results indicate that native A. sturio were feeding on D. polymorpha at a very limited rate and their potential to prevent the establishment of invasive bivalve species, in new and previously invaded areas, is considered limited.     

Microsatellite development for the endangered Yangtze sturgeon (Acipenser dabryanus Duméril, 1869) using 454 sequencing

The Yangtze sturgeon (Acipenser dabryanus) is an endemic species in China. Using 454 sequencing, eight polymorphic tri‐, tetra‐, penta‐, and hexanucleotide microsatellite loci were isolated in this study. The raw sequence data from a one‐eighth run of 454 sequencings were 38.0 Mbp containing 94 222 reads/sequences. Of 80 microsatellite loci, only eight loci were polymorphic in a population of 30 individuals. The number of alleles per locus ranged from 4 to 14 (mean 7.62), and the observed heterozygosities varied between 0.46 and 0.88 (mean 0.74). Cross amplification was tested in congeneric species Acipenser sturio and Acipenser sinensis. These new microsatellite markers will be useful for further studies on genetic variation, parentage analysis, and conservation management for this critically endangered species.     

Assessment of Extinction Risk and Reasons for Decline in Sturgeon

Sturgeon populations in the Danube River have been affected by a combination of hydropower development, over-harvesting, habitat degradation from agricultural and industrial practices and from urbanization. The effects of these changes have been monitored on six sturgeon species inhabiting the Danube River. Two of them are resident species, while the other four migrate to the river for spawning. Atlantic sturgeon (Acipenser sturio) has completely disappeared from this region. Ship sturgeon (Acipenser nudiventris) is very rare in professional fishing catches. Beluga (Huso huso), Russian sturgeon (Acipenser gueldenstaedtii), stellate sturgeon (Acipenser stellatus) and sterlet (Acipenser ruthenus) are endangered with different levels of extinction risk. Here, we model the time dependence of the beluga and Russian sturgeon catch in the Serbian part of the Danube River. Predicted extinction of Russian sturgeon was estimated to fall around the middle of the century, and for beluga approximately at middle of the millennium. Suggestions for sturgeon conservation measures on a national level and coordination of all relevant institutions in Serbia are also presented.     

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.     

Genetic Identification of Western Mediterranean Sturgeons and its Implication for Conservation

To date, the only native sturgeon species in Western Europe was believed to be Acipenser sturio. However, this species is currently restricted to the Gironde River (Southern France), and it poses serious difficulties for rearing in captivity and for using in recovery programme. Furthermore, it has been questioned whether A. sturio is in fact the only species within the rivers of Western Europe, as A. naccarii, a species previously considered endemic of the Adriatic region, has been reported from the Iberian Peninsula in recent years. Here, we have used forensic techniques to obtain DNA from several museum specimens of sturgeons caught in the Spanish Guadalquivir River and in other European rivers. We analysed DNA sequences from these specimens for five genetic markers (three nuclear and two mitochondrial regions), which were subsequently compared with sequences obtained from A. sturioand A. naccarii. Our study demonstrates that A. naccarii coexists withA. sturio, from the Adriatic Sea to the Iberian Peninsula, a finding that could be taken into account in future sturgeon recovery programmes in Western Europe.     

Comparative morphology of the pectoral fin spine of the Persian sturgeon Acipenser persicus,the Russian sturgeon Acipenser gueldenstaedtii,and the Starry sturgeon Acipenser stellatus in Iranian waters of the Caspian Sea

The morphological characteristics of the pectoral fin spine were compared in three species of sturgeon, the Persian sturgeon (Acipenser persicus), the Russian sturgeon (Acipenser gueldenstaedtii), and the Starry sturgeon (Acipenser stellatus), all sampled from the Caspian Sea. On the basis of morphological characters of the pectoral fin spine, 62.2% of the individuals were correctly classified into separate groups. The cluster analysis also divided the three species into two major subgroups. Acipenser persicus and A. gueldenstaedtii were grouped together, suggesting a similar evolutionary basis. Significant morphological heterogeneity in pectoral fin spine characteristics was observed among the three sturgeon species. Principal component analysis identified the largest differences were in the pectoral fin spine size and the angle between distal pectoral fin spine and the horizontal line (A°). The first and second principal components (PC1 and PC2) of all observations accounted for 64.19% and 14.33% of the total variation, respectively. The combination of all analyses showed the relevance of applying pectoral fin spine shape for interspecific distinction of the three species of sturgeons.     

Microsatellite DNA variation in Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) and cross-species amplification in the Acipenseridae

Overharvest and habitat alteration have led toa collapse of most commercial Atlantic sturgeon(Acipenser oxyrinchus oxyrinchus)fisheries while pushing the species to rarityor extirpation in most of its historical range. A biologically sound conservation program forthis species requires knowledge of its geneticdiversity and of the evolutionary relationshipsamong geographic populations. To address theseresearch needs, six microsatellite loci wereisolated from A. o. oxyrinchus. Pedigreeanalysis suggested that all are inherited in acodominant Mendelian pattern. The six lociwere tested in ten additional sturgeon speciesfrom three genera and three apparent ploidylevels (4n, 8n, 16n). Approximately 70% ofsuccessful locus-species amplifications werepolymorphic. Polysomy was observed most oftenin 8n and 16n species. Genetic diversity andpopulation structure of A. o. oxyrinchuswere assayed using three polymorphic Aoxmarkers and four markers developed from lakesturgeon (A. fulvescens). A. o.oxyrinchus were sampled from the AltamahaRiver, Georgia, USA north to the St. LawrenceRiver, Quebec, Canada. Gulf sturgeon, A.o. desotoi, were sampled from the SuwanneeRiver, Florida, USA, to assess differentiationbetween the subspecies. Seventy-seven alleleswere observed to segregate into uniquemultilocus genotypes for each of the 392individuals assayed. Mean diversity wasgreatest in the Chesapeake Bay (9.7 alleles perlocus) and Delaware River (7.4 alleles perlocus) collections, and lowest in the St.Lawrence River (4.6 alleles per locus). Meanheterozygosity across seven loci ranged from44.3% (St. Lawrence River) to 62.6% (Altamaha River). Significant allelic heterogeneity wasobserved in 82% of pairwise comparisons aswell as a global test (p < 0.0001) for A.o. oxyrinchus collections. Genetic distancesuggests the presence of at least sixsubpopulations in A. o. oxyrinchus: St.Lawrence River, St. John River, Hudson River,Delaware River, Albemarle Sound, and AltamahaRiver. Genetic and geographic distances werepositively correlated (r = 0.57, p < 0.03) amongA. o. oxyrinchus, suggesting isolation bydistance and philopatry. Hierarchical genediversity analysis indicated significantgenetic population structure at every level. Maximum likelihood assignment tests correctlyassigned individual fish to collection with ahigh rate of success (mean = 87.5%); this andother lines of evidence indicated that theChesapeake Bay collection represents a mixedpopulation of sub-adult sturgeon from northernand southern Atlantic coast populations. Population structure was correlated with thatsuggested by earlier mitochondrial (mt) DNAanalyses. Significant diversity was observedbetween two Canadian populations from whichonly a single mtDNA haplotype has beenreported.     

Sturgeons in Greece: a review

In the past, sturgeons were practically unknown in Greece, both to the public and to scientists, the latter not having had the opportunity to study wild populations of the four native species. Populations of stellate sturgeon (Αcipenser stellatus Pallas, 1771), Adriatic sturgeon (Acipenser naccarii Bonaparte, 1836), and beluga sturgeon (Huso huso L., 1758) gradually collapsed by the end of the 1970s. Only the River Evros (Thrace, N.W. Greece) sustained a small fishery and caviar canning operation with European sturgeon (A. sturio L., 1758) until 1975. Collapse of stocks was mainly attributed to overfishing, pollution and damming. Sturgeons became widely known after initial farming efforts by the Municipal Hatchery at Lake Ioannina in 1992. Broodstock or fertilized eggs of species with high aquaculture potential, such as sterlet (Acipenser ruthenus L., 1758), Russian sturgeon (Acipenser gueldenstaedtii Brandt, 1833), bester hybrid (A. ruthenus ×H. huso), paddlefish (Polyodon spathula Walbaum, 1792), Siberian sturgeon (Acipenser baerii Brandt, 1869) and white sturgeon (Acipenser transmontanus Richardson, 1836) were imported into Greece between 1992 and 2004. Hatchery technology, larval rearing and production systems are reviewed in this paper and, despite problems, past and present efforts appear to meet a particular interest of the aquaculture sector as well as of those interested in the restoration of wild stocks.     

Induced maturation and spawning in an incidentally caught adult pair of critically endangered European sturgeon, Acipenser sturio L.

Described is how a pair of wild spawners of the critically endangered common Atlantic sturgeon, Acipenser sturio L., were induced to provide progeny despite their unsuitable reproductive states. When captured, the female revealed signs of over‐maturation. The male, caught in brackish water, yielded no sperm; after 5 days in fresh water and with pituitary injection, motile spermatozoa were produced. A batch of 30% fertilized eggs was produced with a two‐injection sequence. As opposed to the Siberian sturgeon, A. baeri, the A. sturio L. hatchlings showed no positive phototaxis.     

A Noninvasive Technique for Determining Sex of Live Adult North American Sturgeons

We have developed a noninvasive technique to determine the sex of adult North American sturgeon by examination of the external morphology of live individuals. We analyzed four sturgeon species taken by commercial and scientific harvest: white Acipenser transmontanus, green A. medirostris, Atlantic A. oxyrinchus and shortnose A. brevirostrum. Males have a urogenital opening in the shape of a letter Y, while the opening of females is in the shape of the letter O. We accurately sexed 26 of 36 sturgeon using this criterion. Accuracy was highest in live fish (82%), and significantly lower in dead fish (29%). Dead sturgeon usually have the rectum prolapsed so that the urogenital opening is protruded, and thus making the sexes indistinguishable.     

Fecundity of Atlantic sturgeon (Acipenser oxyrinchus Mitchill, 1815) captured by the commercial fishery in the Saint John River,New Brunswick,Canada

Knowledge of fecundity in fishes is important for understanding life history, modeling population dynamics, developing fisheries management and determining conservation status. Sturgeon species are known for their high fecundity but to date only two populations of Atlantic sturgeon (Acipenser oxyrinchus Mitchill, 1815) have been examined for this aspect of their biology. The species is protected in most regions of eastern North America but the Saint John River, New Brunswick supports one of the two remaining commercial fisheries in Canada. Sixty females selected by this fishery were sampled for total length, weight, age, gonad weight and egg number. Absolute fecundity ranged from 153,630 to 1,306,626 eggs, with a mean fecundity of 582,832 ± 261,806 eggs. Absolute fecundity exhibited a significant positive relationship to increasing body weight but not to total length or age. Relative fecundity ranged from 129 to 216 eggs/g gonad weight and although it declined with both increasing body size and age the relationships were not significant. The Gonadosomatic Index (GSI) ranged from 1.3% to 27.0% with a mean of 7.8% ± 4.94 SD. Relative fecundity found in this study was higher and mean GSI lower than reported for other Atlantic sturgeon populations and other sturgeon species. Differences were probably due to location of the fishery downstream from spawning sites and selection of pre-spawning females by the commercial fishery.     

Laboratory study of the susceptibility of hatchery reared Atlantic sturgeon (Acipenser oxyrinchus Mitchill, 1815) larvae to predation

The aim of the study was to determine the susceptibility to predation of Atlantic sturgeon larvae (Acipenser oxyrinchus) reared under traditional hatchery conditions. Experiments were conducted to determine whether predators would prey on Atlantic sturgeon if alternative prey was available and if the presence of substrate on the tank bottom impacted the number of Atlantic sturgeon consumed. European perch (Perca fluviatilis) was used as the predator, and the alternative prey were three‐spined stickleback (Gasterosteus aculeatus) or gudgeon (Gobio gobio). The predators and alternative prey were obtained from the wild. The mortality of sturgeon (n = 10) and alternative prey (n = 10) caused by four predators was recorded during 15 min trials. Trials with three‐spined stickleback and gudgeon as alternative prey were performed separately. Each experimental trial was repeated five times. The predators consumed significantly more Atlantic sturgeon than alternative prey in both the experimental setups when the bottom of the tank was covered with gravel and stone substrate and when there was no substrate. In trials with three‐spined stickleback the mortality of Atlantic sturgeon in both experimental setups was 94 ± 8.94%, while that of three‐spined stickleback in the setup with substrate was 20 ± 19.23%, and without substrate it was 22 ± 10.00%. European perch also consumed more Atlantic sturgeon than they did gudgeon, and the mean Atlantic sturgeon mortality in the experimental setup with substrate was 94 ± 5.48%, while for gudgeon it was 48 ± 8.37%. In the experimental setup without substrate the predators also consumed substantially more Atlantic sturgeon than gudgeon, with a mean Atlantic sturgeon mortality of 94 ± 8.94%, while for gudgeon it was 76 ± 5.48%. The study indicated that hatchery reared Atlantic sturgeon larvae are susceptible to predation by European perch. Predation could impact the survival of juvenile Atlantic sturgeon in the natural environment, and it could be one of the factors that is impeding the restoration of this species in the Baltic Sea basin.     

From microsatellites to single nucleotide polymorphisms for the genetic monitoring of a critically endangered sturgeon

The use of genetic information is crucial in conservation programs for the establishment of breeding plans and for the evaluation of restocking success. Short tandem repeats (STRs) have been the most widely used molecular markers in such programs, but next‐generation sequencing approaches have prompted the transition to genome‐wide markers such as single nucleotide polymorphisms (SNPs). Until now, most sturgeon species have been monitored using STRs. The low diversity found in the critically endangered European sturgeon (Acipenser sturio), however, makes its future genetic monitoring challenging, and the current resolution needs to be increased. Here, we describe the discovery of a highly informative set of 79 SNPs using double‐digest restriction‐associated DNA (ddRAD) sequencing and its validation by genotyping using the MassARRAY system. Comparing with STRs, the SNP panel proved to be highly efficient and reproducible, allowing for more accurate parentage and kinship assignments' on 192 juveniles of known pedigree and 40 wild‐born adults. We explore the effectiveness of both markers to estimated relatedness and inbreeding, using simulated and empirical datasets. Interestingly, we found significant correlations between STRs and SNPs at individual heterozygosity and inbreeding that give support to a reasonable representation of whole genome diversity for both markers. These results are useful for the conservation program of A. sturio in building a comprehensive studbook, which will optimize conservation strategies. This approach also proves suitable for other case studies in which highly discriminatory genetic markers are needed to assess parentage and kinship.     

Interspecific hybridization in natural sturgeon populations of the Eastern Black Sea: the consequence of drastic population decline?

The eastern part of the Black Sea and its tributaries are suitable habitats for several sturgeon species, among which Acipenser gueldenstaedtii, A. stellatus, A. nudiventris, A. persicus, A. sturio, and H. huso are well documented. However, different threats have led these species to a dramatic decline, all of them are currently listed as Critically Endangered, and some Locally Extinct, in that area. We tested 94 wild sturgeon samples from the Black Sea and Rioni River by analyzing the mitochondrial Control Region and nuclear markers for hybrid identification. The data analyses (1) assessed mitochondrial diversity among samples, (2) identified their species, as well as (3) indicated instances of hybridization. The data collected, besides confirming a sharp decrease of catches of Beluga and Stellate sturgeon in recent years, also revealed four juvenile hybrids between Russian and Stellate sturgeon, providing the first evidence of natural interspecific hybridization in the Rioni. The present communication raises concerns about the status of sturgeon species in this area and underlines the urgent need for conservation programs to restore self-sustaining populations.