Analysis of creatine kinase variation in some members of the family Acipenseridae

Creatine kinase (E.C. 2.7.3.2) was examined in stellate sturgeon Acipenser stellatus Pallas, Russian sturgeon A. gueldenstaedtii Brandt, European sterlet A. ruthenus L., Siberian sterlet A. ruthenus marsiglii Brandt, and great sturgeon (beluga) Huso huso L., using polyacrylamide gel electrophoresis. Two loci for creatine kinase were identified: CK-A* in white skeletal muscle and CK-C* in stomach wall muscle. Most species proved to be monomorphic at the CK-A* locus, showing the same phenotype represented by a single band. Heterogeneity and polymorphism in creatine kinase, determined by the CK-A* locus, were found only in Russian sturgeon. Based on the results of densitometric analysis of band staining intensity, we have advanced a hypothesis that synthesis of subunits of the CK-A* product in this species was controlled by eight genes. However, the genotype frequencies in the sample were significantly different from those theoretically expected upon free and independent gene recombination. The results of this study support the hypothesis on the absence of heterodimeric creatine kinase molecules in the skeletal muscle of Russian sturgeon. Locus CK-C* in stellate sturgeon was revealed as a single, intensely stained, rapidly migrating fraction, whereas in Russian sturgeon, the enzyme activity in this zone was very weak. No creatine kinase was found in liver, kidneys, spleen, heart, and intestine mucous tunic.     

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.     

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.     

Factors influencing the natural reproduction decline in the beluga (Huso huso,Linnaeus, 1758), Russian sturgeon (Acipenser gueldenstaedtii,Brandt & Ratzeburg, 1833), and stellate sturgeon (A. stellatus,Pallas, 1771) of the Volga–Caspian basin: A review

The review describes the changes in natural reproduction of three important sturgeon species in the Volga–Caspian basin: (a) the beluga (Huso huso Linneaus, 1758), (b) the Russian sturgeon (Acipenser gueldenstaedtii Brandt &Ratzeburg, 1833), and (c) the stellate sturgeon (Acipenser stellatus Pallas, 1771). Since the past 60 years, these species responded to severe influences of natural and anthropogenic factors. On the basis of original and published data, an analysis has been made of (a) the numbers of larvae migrating from spawning sites (according to plankton net survey), (b) fecundity and histological anomalies in gonad development, (c) the numbers of adult sturgeons in the Caspian Sea and of spawners migrating to the Volga River (according to trawl and beach seine survey), and (d) foraging resources for the sturgeons. The results show that their natural reproduction in the Volga–Caspian basin has declined drastically during the past decades under the impact of (a) fluctuations of the Caspian Sea level and flow discharge from the Volga River, (b) blockage of sturgeon migration routes and loss of spawning sites because of dam construction, (c) water pollution in the lower reaches of the Volga River and in the Caspian Sea, and (d) intensive and selective illegal and unreported fishing. The relative significance of these factors has been changing during the study period.     

Morphological peculiarities of pronephros in Russian sturgeon, Acipenser gueldenstaedtii Brandt (order acipenseriformes, family acipenseridae)

The structure of the pronephros in Russian sturgeon larvae, Acipenser gueldenstaedtii Brandt, at different stages of early postembryonic development (from hatching till 14 days old), was studied with histological and electron microscopy methods. The formed pronephros is represented by a system of bilaterally located pronephric tubules and an external single glomus, which is not integrated directly into pronephric tubules and is located in closed pronephric chamber. The glomus is positioned below the dorsal aorta and is vascularized by its capillaries. The thin structure of the glomus has the same characteristic features that are typical of and needed for the functions of any filtering organ. By the time when larvae transfer fully to exogenous feeding, the pronephros undergoes significant degradation and it is replaced by the mesonephric kidney which develops during the period of function of the pronephros. The peculiarities of the pronephros in acipenserids are discussed comparatively with the same organ in teleosts and amphibians.     

Comparative enzymologic study of catalytical properties of liver monoamine oxidases of sturgeon fish

We carried out the comparative study of the substrate and inhibitory specificity of liver monoamine oxidases (MAO) of the giant sturgeon Huso huso, the starred sturgeon Acipenser stellatus, the Persian sturgeon Acipenser persicus, and the Russian sturgeon Acipenser gueldenstaedtii. Results of the substrate-inhibitor analysis with use of inhibitors chlorgilin and deprenil, as well as five specific substrates indicate homogeneity of these enzymes. All studied MAO have the several orders higher sensitivity to chlorgilin than to deprenil, with essential interspecies differences being observed. There are determined kinetic parameters of enzymatic deamination (K _M and V) of tyramine, serotonin, noradrenalin, benzylamine, β-phenylethylamine, and N-methylhistamine. All studied enzymes have been established to have the higher activity toward serotonin and noradrenalin-substrates of the MAO A form as compared with benzylamine, β-phenylethylamine, and N-methylhistamine-substrate of the mammalian MAO B form, the maximal activity being characteristic of the giant sturgeon.     

Formation of populations of Acipenseridae sturgeons in the Volga-Caspian basin

Analysis of dynamics of commercial catches and the state of populations of Russian sturgeon Acipenser gueldenstaedtii, starred sturgeon A. stellatus, and great sturgeon A. huso (Huso huso) in the Caspian Sea is presented. In the retrospective, the role of natural and artificial reproduction in formation of populations of Acipenseridae sturgeons is considered. The main principles of rational organization of practical activity for preservation and restoration of populations of Acipenseridae sturgeons are formulated.     

Reaction of osmoregulatory system in juveniles of Russian sturgeon <Emphasis Type="Italic">Acipenser gueldenstaedtii</Emphasis> (Acipenseridae) on influence of comatotropin of carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis>

For the first time by the example of juveniles of Russian sturgeon Acipenser gueldenstaedtii Brandt, the role of growth hormone of hypophysis in the osmotic regulation of acipenserids was studied, also the process of transformation of reserve chloride was described. In Russian sturgeon juveniles, administration of growth hormone of carp Cyprinus carpio at a dose of 1 mg per 1 g body weight causes changes in the fine structure of numerous reserve (undifferentiated) gill epithelial cells and thus stimulates their transformation into mature chloride cells involved in transport of monovalent ions. Increase in the number of chloride cells, on account of reserve cells, was accompanied by higher enzymatic activity of Na⁺/K⁺-ATPase in gill homogenates of the injected fish adapting to a hyperosmotic environment of salinity 12.5‰ (403 mosm/l) than in the intact fish under the same conditions but without hormonal treatment. Morphophysiological changes observed in the injected juveniles provided the more efficient dynamics of blood serum osmolarity under saline exposure than that in the intact fish. Their serum osmolality during the maximum rise (24 hours after the transfer of fish into the water of 12.5‰ salinity) was lower (p <0.01), than that of the intact fish. The inclusion of growth hormone in the osmoregulatory process was established.     

Discrimination of interspecific hybrids in natural populations of Amur sturgeon fish by means of multilocus RAPD-PCR markers

A RAPD-PCR analysis of a sample of Amur sturgeon fish (46 individuals) is performed. An estimate of the genetic state of native populations of Amur sturgeon Acipenser schrenckii (Brandt, 1869) and kaluga fish Huso dauricus (Georgi, 1775) is given. Genetic evidence for the hybrid origin of two phenotypical hybrids is obtained. Estimates of the genetic distances between the species and the hybrids proved to be at the level of interspecific distances. It is shown that an Exact Test for differentiation of the populations is the most effective means of discriminating the two species, while multidimensional scaling (MDS) is the most effective means of discriminating the hybrids. It is concluded that the populations of sturgeon fish in the Amur River have maintained a substantial level of genetic diversity; the presence of hybrids in these populations is judged to be one of the risk factors. Multilocus RAPD-PCR markers are acknowledged as a convenient and reliable tool for conducting genetic monitoring of populations of Amur sturgeon fish for the purpose of preserving the gene pool of the fish.     

Biological peculiarities of the industrially bred juvenile Russian sturgeon Acipenser gueldenstaedtii (Acipenseridae, Acipenseriformes) in the Don

We discuss results of the investigation into biological peculiarities of the juvenile Russian sturgeon (Acipenser gueldenstaedtii) from the Aksay-Don Sturgeon Hatchery living in a natural riverine environment and migrating seaward. We supply data on the growth rate, geographic distribution, behavior and the feeding manner of the juvenile Russian sturgeon in the Don. We have estimated the survival rate of the industrially bred juvenile Russian sturgeon living in a natural environment. Apart from the nutritive base in the river and the predation pressure, we found that ecologically and physiologically determined biological characteristics mostly influence the survival rate in the juvenile Russian sturgeon released into the Don. The findings allow estimation of artificial propagation of the Russian sturgeon in the Azov Sea basin.     

Analysis of creatine kinase variation in some members of the family Acipenseridae

Creatine kinase (E.C. 2.7.3.2) was examined in stellate sturgeon Acipenser stellatus Pallas, Russian sturgeon A. gueldenstaedtii Brandt, European sterlet A. ruthenus L., Siberian sterlet A. ruthenus marsiglii Brandt, and great sturgeon (beluga) Huso huso L., using polyacrylamide gel electrophoresis. Two loci for creatine kinase were identified: CK-A* in white skeletal muscle and CK-C* in stomach wall muscle. Most species proved to be monomorphic at the CK-A* locus, showing the same phenotype represented by a single band. Heterogeneity and polymorphism in creatine kinase, determined by the CK-A* locus, were found only in Russian sturgeon. Based on the results of densitometric analysis of band staining intensity, we have advanced a hypothesis that synthesis of subunits of the CK-A* product in this species was controlled by eight genes. However, the genotype frequencies in the sample were significantly different from those theoretically expected upon free and independent gene recombination. The results of this study support the hypothesis on the absence of heterodimeric creatine kinase molecules in the skeletal muscle of Russian sturgeon. Locus CK-C* in sterlet was revealed as a single, intensely stained, rapidly migrating fraction, whereas in Russian sturgeon, the enzyme activity in this zone was very weak. No creatine kinase was found in liver, kidneys, spleen, heart, and intestine mucous tunic.     

A biometric method for determining the sex of acipenserids, including the Russian sturgeon Acipenser gueldenstaedtii (Acipenseridae) of the Azov Population

With reference to the Russian sturgeon Acipenser gueldenstaedtii of the Azov Population, it is demonstrated that the sex of acipenserids may be determined biometrically. A measurement scheme of the head is suggested, which gives the possibility to determine gender in live sturgeons. A discriminate equation is composed for the identification of the sex of individuals of the Russian sturgeon in a sufficiently wide size-age range, irrespective of the maturity stage of the gonads.     

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.     

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.     

Development of mesonephros in prolarvae of acipenserids (acipenseridae)

Development of the excretory system in early ontogenesis of acipenserids—the Russian sturgeon Acipenser gueldenstaedtii, the starred sturgeon A. stellatus, the sterlet A. ruthenus, and the great sturgeon Huso huso—is investigated histologically. In acipenserids, the mesonephros develops after hatching. At the 36th stage of development (after Detlaf et al., 1981), the mesonephros is a consecutively arranged group of spherical rudiments of placodes situated along the Wolffian duct. Morphological differentiation of mesonephros as the first generations of renal corpuscles, vascular glomeruli, and convoluted tubules is completed up to the 45th stage, i.e., to the moment of transition of prolarvae to active feeding.     

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.     

Distribution of the enzyme activity in the intestine of beluga <Emphasis Type="Italic">Huso huso</Emphasis> and Russian sturgeon <Emphasis Type="Italic">Acipenser gueldenstaedtii</Emphasis> (Acipenseridae)

Data on distribution of digestive enzymes along the intestinal tract of mature specimens of the beluga Huso huso and Russian sturgeon Acipenser gueldenstaedtii are presented. The absence of a distinct proximo-distal gradient of the enzyme activity can be explained by short length of the intestine of sturgeons. Considerable differences are found in the distribution pattern of enzymes along the intestinal tract of beluga and Russian sturgeon. The effect of the method of calculation of the enzyme activity on the data on the proximo- distal gradient is considered.     

Comparison of the Effects of Gonadotropic Preparations of the Carp and Stellate Sturgeon Pituitaries onin vivoand in vitroOocyte Maturation in the Siberian Sturgeon Acipenser baeriBrandt

Injections of 2.5 mg/kg of stellate sturgeon (Acipenser stellatusPall.) pituitary extract and 5 mg/kg of carp (Cyprinus carpioL.) pituitary extract in Siberian sturgeon (Acipenser baeriBrandt) females did not reveal significant differences in the effects of these preparations. There were no differences in the percentage of females that responded by ovulation, duration of the period from injection to ovulation, rate of ovulation, or quality of mature eggs as estimated by the percentage of fertilization or percentage of normal embryos at the small yolk plug stage. Thus, an insufficient efficiency in the artificial reproduction of the Siberian sturgeon grown in captivity is not related to the use of the carp pituitary preparation as a stimulus. Estimation of the ratio of specific activities of the pituitary extracts and purified gonadotropins of the stellate sturgeon and carp by in vitrooocyte maturation has shown that it varies within wide limits as a function of the medium composition and physiological state of follicles. Hence, the ratio of activities of the gonadotropins of different species as determined by in vitromaturation of sturgeon oocytes may markedly differ from that upon injection of these preparations in breeders.     

Molecular identification and the features of genetic diversity in interspecific hybrids of Amur sturgeon ( Acipenser schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus , and A. ruthenus × A. schrenckii ) based on variability of multilocus RAPD markers

The method of polymerase chain reaction with random primers (RAPD PCR) was used to identify the progeny of the crosses between three sturgeon species, Amur sturgeon (Acipenser schrenckii Brandt, 1869), Siberian sturgeon (A. baerii Brandt, 1869), and sterlet (A. ruthenus Linnaeus, 1758). Using ten primers, genetic variation in 70 yearlings, produced in seven individual crosses: Acipenser schrenckii × A. schrenckii, A. baerii × A. baerii, A. ruthenus × A. ruthenus, A. schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus, and A. ruthenus × A. schrenckii was described and evaluated. It was demonstrated that the samples composed of hybrids from individual crosses were more variable than the samples of parental species. On the other hand, pooled samples of hybrids from two cross directions were genetically less variable than the pooled samples of their parents. The three main features of the hybrid RAPD profiles identified included: (1) preservation of marker DNA fragments of both parents in one genome; (2) presence of specific DNA fragments, absent from both parents; and (3) dependence of the frequency of some DNA fragments from the cross direction. Multidimensional scaling clearly distinguishes in the space of three coordinates the individuals of original species and the hybrid progeny with differentiation in the groups of direct and backcross hybrids. Analysis of relationships (UPGMA and NJ) pointed to substantial differentiation between the species, as well as between the species and hybrid progeny. Close genetic relationships between direct and backcross hybrids were demonstrated. Multilocus RAPD markers in association with statistical methods are considered to be the useful tool for discrimination of interspecific hybrids of sturgeon. Possible reasons for the differences in the hybrid RAPD profiles are discussed. Original Russian Text ? K.V. Rozhkovan, G.N. Chelomina, E.I. Rachek, 2008, published in Genetika, 2008, Vol. 44, No. 11, pp. 1453–1460.     

High‐throughput SNP‐genotyping analysis of the relationships among Ponto‐Caspian sturgeon species

Legally certified sturgeon fisheries require population protection and conservation methods, including DNA tests to identify the source of valuable sturgeon roe. However, the available genetic data are insufficient to distinguish between different sturgeon populations, and are even unable to distinguish between some species. We performed high‐throughput single‐nucleotide polymorphism (SNP)‐genotyping analysis on different populations of Russian (Acipenser gueldenstaedtii), Persian (A. persicus), and Siberian (A. baerii) sturgeon species from the Caspian Sea region (Volga and Ural Rivers), the Azov Sea, and two Siberian rivers. We found that Russian sturgeons from the Volga and Ural Rivers were essentially indistinguishable, but they differed from Russian sturgeons in the Azov Sea, and from Persian and Siberian sturgeons. We identified eight SNPs that were sufficient to distinguish these sturgeon populations with 80% confidence, and allowed the development of markers to distinguish sturgeon species. Finally, on the basis of our SNP data, we propose that the A. baerii‐like mitochondrial DNA found in some Russian sturgeons from the Caspian Sea arose via an introgression event during the Pleistocene glaciation.