Site heteroplasmy in the mitochondrial cytochrome b gene of the sterlet sturgeon Acipenser ruthenus

Sturgeons are fish species with a complex biology. They are also characterized by complex aspects including polyploidization and easiness of hybridization. As with most of the Ponto-Caspian sturgeons, the populations of Acipenser ruthenus from the Danube have declined drastically during the last decades. This is the first report on mitochondrial point heteroplasmy in the cytochrome b gene of this species. The 1141 bp sequence of the cytb gene in wild sterlet sturgeon individuals from the Lower Danube was determined, and site heteroplasmy evidenced in three of the 30 specimens collected. Two nucleotide sequences were identified in these heteroplasmic individuals. The majority of the heteroplasmic sites are synonymous and do not modify the sequence of amino acids in cytochrome B protein. To date, several cases of point heteroplasmy have been reported in animals, mostly due to paternal leakage of mtDNA. The presence of specific point heteroplasmic sites might be interesting for a possible correlation with genetically distinct groups in the Danube River.     

Acipenser sturio and Acipenser nudiventris in the Danube – extant or extinct?

The presence of Atlantic sturgeon ( Acipenser sturio) and ship sturgeon ( Acipenser nudiventris ) in the Danube has been known through only occasional, chance sightings; significant uncertainty remains as to whether these species are already extinct or still present in the Danube River basin. In this study, five different statistical methods were applied for assessment of their extinction probability. All methods provided a significant probability that the Atlantic sturgeon is extinct, with extinction having occurred somewhere between 1966 and 1970. Although the applied methods provided different results, all indicated that the ship sturgeon is probably still present, but that extinction may occur within a few decades. Sensitivity analyses of these statistical methods projected that new sightings of ship sturgeon in the coming years would not extend the length of extinction by more than a few years. The best management approach for these two species, given the lack of knowledge on their presence in the Danube River basin, could be to apply all feasible protection measures for the other four Danube sturgeon species. These measures could at the same time provide protection for these two species, presuming they are not already extinct or beyond recovery.     

Past and current status of sturgeons in the upper and middle Danube River

Of the six species of sturgeons native to the Danube basin, five occurred in the upper and middle Danube. Among anadromous sturgeons were the large winter races of beluga, Huso huso, Russian sturgeon, Acipenser gueldenstaedtii, and stellate sturgeon, A. stellatus, which ascended the middle, and sometimes also the upper Danube, to spawn. Due to overfishing, followed by severe habitat alteration including damming and pollution, these anadromous sturgeons are critically endangered or extirpated from the upper and middle Danube. Acipenser gueldenstaedtii and A. nudiventris are represented only as resident non-migratory races with very small populations. The most abundant and widely distributed species is the sterlet, A. ruthenus, although it is presently limited to the middle Danube. Its population increased in some sections of the middle Danube during the past 15 years, presumably because of improving water quality, but this species remains at risk because of continuing habitat degradation.     

A Comment on the Siberian, Acipenser baerii, and Russian, Acipenser gueldenstaedtii, Sturgeons

Contrary to the opinion of Kynard et al. (2002), the Siberian sturgeon, Acipenser baerii, does not belong to the Ponto-Caspian species. It inhabits Siberian rivers and Lake Baikal. Acipenser baerii is a typical potamodromous species and the comparison of the behavior of its embryos and larvae with those of the anadromous Russian sturgeon, Acipenser gueldenstaedtii, should be done with understanding that these species have different life histories. The statement by Kynard et al. (2002) that larvae of the Russian sturgeon do not migrate contradicts results of previous studies.     

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.     

Comparative analysis of morphometric characters of juvenile sterlet Acipenser ruthenus L. from natural population and aquaculture

A comparative analysis was performed on sixteen morphometric characters, in three different juvenile sterlet Acipenser ruthenus L. populations. Specimens were collected from a wild population in the Serbian part of the Danube River (n = 46), from aquaculture stocks in the Czech Republic originating from Russia (n = 40), and aquaculture stocks in the Slovak Republic originating from Slovakian part of the Danube River (n = 28). Average values for total length were 29·9 ± 3·9 cm, 29·1 ± 3·7 cm and 27·3 ± 7·7 cm for Serbia, Czech Republic and Slovak Republic, respectively. Populations were compared using t‐test and sequential Bonferroni correction for multiple comparisons was applied in order to determine significant differences between them. Results of analysis showed that all three populations differed in prebarbel length, interocular distance and maximum head width. Although all these characters are head‐related, head length itself was very uniform among all populations. The natural population from the Serbian part of the Danube River differed from the populations reared in aquaculture in seven morphometric characters. The two populations reared in aquaculture consistently showed lower morphological variability than the wild population, even though they had different genetic backgrounds (Russia and Slovakia). Future genetic studies will show if this tendency is caused by a reduction in genetic variability.     

Morphometric recognition of two morphs in sterlet (Acipenser ruthenus) population induced by different reproductive behaviour

Four species of sturgeons appear in the Yugoslav part of the Danube River. Among them three are anadromous, beluga ( Huso huso ), Russian sturgeon ( Acipenser gueldenstaedtii ) and stellate sturgeon ( Acipenser stellatus ) and migrate to Danube from the Black Sea to spawn. Only sterlet ( Acipenser ruthenus ) is exclusively freshwater. There are obvious differences between vernal and winter races of anadromous sturgeons, due to their different migratory patterns. Russian ichthyologist Berg (1934) claimed that there are two races (morphs) in the starlet population as well. This stimulated long discussion in Russian and Yugoslav literature, and it was the starting point for our research on sterlet in the Yugoslav part of the Danube River.
Morphometric analysis was preformed on 159 individuals caught in Danube River among 1123 and 1173 km, on four localities during September and October 2002. Analysis included 24 morphological and 3 meristic characters. K‐means cluster analysis indicated existence of two different clusters (groups), one with 95 (59·75%) and other with 64 (40·25%) samples. Significant differences were found for preorbital, preoral and prebarbel length. Mean values (in % of head length) for first group were 50·13 ± 1·71, 57·61 ± 1·87, 39·74 ± 2·20 and for second group 53·50 ± 2·46, 62·07 ± 2·20 and 43·30 ± 2·08 respectively. Findings were confirmed by Principal Components & Classification Analysis. These results speak in favour of existence of two morphs in sterlet population, one with long and pointed rostrum (vernal) and other with short rostrum (winter). According to the literature that is probably influenced by different reproductive behaviour (time separation in spawning).     

Natural genetic polymorphism and phylogeography of Siberian sturgeon <Emphasis Type="Italic">Acipenser baerii</Emphasis> Brandt, 1869

Five populations of Siberian sturgeon Acipenser baerii collected throughout the species distribution range (Lake Baikal, Lena, Yenisei, Kolyma, and Irtysh rivers) were examined for genetic polymorphism using five tetraploid microsatellite loci and the mitochondrial DNA control region. It was demonstrated that Siberian sturgeon was represented by genetically well-differentiated groups that corresponded to hydrographic basins, Ob–Irtysh, Baikal–Yenisei, Lena, and Kolyma. Population assignment of the Siberian sturgeon broodstock should be considered in the restocking programs.     

Features of inheritance of several morphological characters in the hybrids between Siberian sturgeon <Emphasis Type="Italic">Acipenser baerii</Emphasis> and beluga <Emphasis Type="Italic">A. huso</Emphasis> (Acipenseridae) and in gynogenetic individuals of Siberian sturgeon

Comparative analysis of several morphological characters is conducted in experimentally obtained hybrids between Siberian sturgeon Acipenser baerii and beluga A. huso, in parental species, and in gynogenetic progeny of Siberian sturgeon. The gynogenetic individuals are similar to Siberian sturgeon by all characters used for differentiation between Siberian sturgeon and beluga that supports gynogenetic origin of these individuals. Based on the data obtained on the hybrid specimens, the features of inheritance of the studied characters in sturgeons and their variation in postlarval ontogeny are discussed.     

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.     

Spawning by lake sturgeon (Acipenser fulvescens) in the Detroit River

Overfishing and habitat destruction in the early 1900s devastated lake sturgeon (Acipenser fulvescens) populations in the Great Lakes. Although a comprehensive restoration strategy for this species was recently drafted by the Michigan Department of Natural Resources, a lack of current data on Great Lakes sturgeon stocks has hindered rehabilitation efforts. Historically, the Detroit River supported one of the largest lake sturgeon populations in the Great Lakes; however, little is known about the current population or its habitat use. The main objective of this study was to determine if lake sturgeon spawns in the Detroit River. As part of a larger study, baited setlines were used to capture lake sturgeon in the Detroit River in the spring and summer of 2000 and 2001. In each year of the study, ultrasonic transmitters were surgically implanted in 10 adult fish to track their movements, evaluate habitat use and identify possible spawning sites. Using telemetry and egg mats to verify spawning activity, one spawning site was located and verified in the Detroit River. Spawning was verified by recovering sturgeon eggs deposited on egg collection mats anchored at the site. Telemetry data suggested that several other possible spawning sites also may exist, however, spawning activity was not verified at these sites.     

Genetic assessment of lake sturgeon (<Emphasis Type="Italic">Acipenser fulvescens</Emphasis>) population structure in the Ottawa River

Lake sturgeon (Acipenser fulvescens) are of conservation concern throughout their range. Many populations are dependent on fluvial habitats which have been increasingly impacted and fragmented by dams and human development. Although lake sturgeon were once abundant in the Ottawa River and its tributaries, historical commercial harvests and other anthropogenic factors caused severe declines and low contemporary numbers in lake sturgeon populations. Contemporary habitat fragmentation by dams may be increasing isolation among habitat patches and local rates of decline, raising concerns for persistence of local populations. We used microsatellite DNA markers to assess population structure and diversity of lake sturgeon in the Ottawa River, and analyzed samples from 10 sites that represent more than 500 km of riverine habitat. To test for evidence of anthropogenic fragmentation, patterns of genetic diversity and connectivity within and among river segments were tested for concordance with geographic location, separation by distance and obstacles to migration, considering both natural and artificial barriers as well as barrier age. Despite extensive habitat fragmentation throughout the Ottawa River, statistical analyses failed to refute panmixia of lake sturgeon in this system. Although the long generation time of lake sturgeon appears to have effectively guarded against the negative genetic impacts of habitat fragmentation and loss so far, evidence from demographic studies indicates that restoring connectivity among habitats is needed for the long-term conservation and management of this species throughout this river system.     

The effects of population and thermal acclimation on the growth,condition and cold responsive mRNA expression of age-0 lake sturgeon (Acipenser fulvescens)

In Manitoba, Canada, wild lake sturgeon (Acipenser fulvescens) populations exist along a latitudinal gradient and are reared in hatcheries to bolster threatened populations. We reared two populations of lake sturgeon, one from each of the northern and southern ends of Manitoba and examined the effects of typical hatchery temperatures (16°C) as well as 60-day acclimation to elevated rearing temperatures (20°C) on mortality, growth and condition throughout early development. Additionally, we examined the cold shock response, which may be induced during stocking, through the hepatic mRNA expression of genes involved in the response to cold stress and homeoviscous adaptation (HSP70, HSP90a, HSP90b, CIRP and SCD). Sturgeon were sampled after 1 day and 1 week following stocking into temperatures of 8, 6 and 4°C in a controlled laboratory environment. The southern population showed lower condition and higher mortality during early life than the northern population while increased rearing temperature impacted the growth and condition of developing northern sturgeon. During the cold shock, HSP70 and HSP90a mRNA expression increased in all sturgeon treatments as stocking temperature decreased, with higher expression observed in the southern population. Expression of HSP90b, CIRP and SCD increased as stocking temperature decreased in northern sturgeon with early acclimation to 20°C. Correlation analyses indicated the strongest molecular relationships were in the expression of HSP90b, CIRP and SCD, across all treatments, with a correlation between HSP90b and body condition in northern sturgeon with early acclimation to 20°C. Together, these observations highlight the importance of population and rearing environment throughout early development and on later cellular responses induced by cold stocking temperatures.     

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.

     

Comparative coelomic fluid composition of sterlet sturgeon Acipenser ruthenus Linnaeus, 1758, Siberian sturgeon Acipenser baerii Brandt, 1869, and Russian sturgeon Acipenser gueldenstaedtii Brandt & Ratzeburg, 1833

The objectives of this study were to compare the physicochemical properties of coelomic fluid (CF) in three sturgeon species, sterlet sturgeon Acipenser ruthenus (age 5–8 years), Siberian sturgeon Acipenser baerii (age 15–20 years), and Russian sturgeon Acipenser gueldenstaedtii (age 13–18 years). For the study, CF was collected by plastic pipette from the eggs of mature female sterlet sturgeon (N = 10), Siberian sturgeon (N = 7) and Russian sturgeon (N = 4); osmolality, pH, ionic composition (Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl^?), glucose, cholesterol and total protein of the CF were then analyzed. Mean concentration of K⁺ (4.4–6.11 mmol/L), Cl^? (89.8–98.0 mmol/L), Ca²⁺ (0.87–0.96 mmol/L), Mg²⁺ (0.57–0.67 mmol/L), cholesterol (0.13–0.16 mmol/L), total protein (2.41–3.57 g/L), pH (7.92–7.98), and osmolality (190–213 mOsm/kg) of CF were not significantly different among these three species. However, the mean concentration of Na⁺ was significantly lower in sterlet sturgeon (104.6 ± 7.7 mmol/L; p < .05) than in the Siberian (126.4 ± 6.2) and Russian (123.0 ± 5.9) sturgeon. For these three species, Na⁺, K⁺, Cl^?, and Mg²⁺ were the dominating ions; several significant correlations were observed among these ions and other physiochemical properties of CF. This is the first report on the ionic and biochemical composition of the CF of sterlet, Siberian and Russian sturgeon, which can be used as a reference point for further development of artificial media for the short‐term storage of unfertilized sturgeon eggs as well as for the standardization of the fertilization protocol for these species in controlled reproduction.     

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.     

Characterization,tissue distribution of resistin gene and the effect of fasting and refeeding on resistin mRNA expression in Siberian sturgeon (Acipenser baerii)

Resistin as an adipokine identified from rodents in 2001 is involved in many biological processes. However, little is known about this gene in fish. We cloned Siberian sturgeon (Acipenser baerii) resistin cDNA of 795 base pairs, encoding 107 amino acids, which showed 38–40% identity to human and rodents. Real-time quantitative PCR showed that the resistin gene was widely distributed in tissues of Siberian sturgeon, with the highest expression in liver. After fasting for 1, 3, 6 and 10 days, the expression of the resistin gene in the liver of Siberian sturgeon decreased significantly, and after refeeding on the 10 days of fasting the resistin mRNA expression increased rapidly, suggesting that resistin may play an important role in liver in response to starvation. Taken together, these results suggest that resistin may be involved in the regulation of energy homeostasis in liver.     

Assessment of the seasonal usage of the lower Pascagoula River estuary by Gulf sturgeon (Acipenser oxyrinchus desotoi)

The Pascagoula watershed likely offers the greatest possibility for the survival of the Gulf sturgeon, Acipenser oxyrinchus desotoi, within Mississippi. Thus, understanding and preserving the connectivity between distant habitats in this region plays a major role in protecting and managing such anadromous fish populations. The focus of this project was to determine the within‐river routes Gulf sturgeon take through the lower Pascagoula River downstream of the point where it splits (river kilometer 23) into two distinct distributaries. Sixty days were sampled throughout a two‐year period with a total effort of 81 947 net‐meter‐hours and eight Gulf sturgeon were captured, ranging from 74 to 189 cm FL and weighing from 3.6 to 52.6 kg. Using an array of automated telemetry receivers, acoustically tagged Gulf sturgeon movements were monitored within the lower river and associated estuary. Estimated residence times (days) suggest Gulf sturgeon appear to prefer the eastern distributary upriver from Bayou Chemise as the primary travel corridor between freshwater habitats and marine feeding grounds. The western distributary mouth was more highly used by Gulf sturgeon during both seasonal migrations between upriver and offshore habitats. Thus, the western distributary appears to represent the main entrance point utilized by Gulf sturgeon to the Pascagoula River watershed and should be protected as the eastern distributary mouth has been altered from a natural marsh edge to one of hardened surfaces.     

Recovery plan for Kootenai River white sturgeon (Acipenser transmontanus)

The white sturgeon ( Acipenser transmontanus ) of the Kootenai River was listed as endangered on September 6, 1994 by the United States Fish and Wildlife Service. This transboundary population, residing in Kootenay Lake and Kootenay River in Canada, and the Kootenai River in the US, has been in general decline since the mid-1960's. There has been very little recruitment to this population in the last 20 years.
This population became isolated from other white sturgeon populations of the Columbia River basin during the last ice age of approximately 10,000 years ago. The population adapted to the pre-development conditions of the Kootenai system, with a high spring freshet and extensive side channel and low-lying delta marshlands. Modification of the Kootenai River by human activities, such as industrial developments, floodplain dyking, and dam construction has changed the hydrograph of the Kootenai River, altering sturgeon spawning, incubation and rearing habitats and reducing overall biological productivity.
A Kootenai River white sturgeon draft recovery plan was prepared by the US Fish and Wildlife Service in cooperation with other agencies in the US and Canada. The plan was peer reviewed and there was a parallel public consultation process, where public commentary was invited from both sides of the international border. The short-term recovery objectives of the recovery plan are to prevent extinction and re-establish successful natural recruitment. The identified long-term objectives are the re-establishment of a self sustaining population and the restoration of productive habitat, in order to downlist to threatened status and subsequently delist this population when recovery is well established. Specific actions needed for recovery include spring flow augmentation during the reproduction period; a conservation aquaculture program to prevent near-term extinction; habitat restoration, and research and monitoring programs to evaluate recovery progress.     

Induction of meiotic gynogenesis in ship sturgeon Acipenser nudiventris using UV-irradiated heterologous sperm

Diploid gynogenesis was induced in ship sturgeon Acipenser nudiventris using UV-irradiated sperm from Siberian sturgeon Acipenser baerii. The optimal condition for the retention of the second polar body in ship sturgeon was determined to be 10 min after activation/fertilization in experiments. The temperature of cold shock and its duration were 2.5 °C and 30 min, respectively. A total of 30 gynogens of known parentage from experimental treatments were screened using microsatellite DNA analysis, and uniparental transmission in meiogens was confirmed. The results show that heterologous Siberian sturgeon sperm is applicable as UV-irradiated sperm for the induction of gynogenesis in ship sturgeon. This technique may recover the critically endangered sturgeon species that are becoming extinct.