Heterogeneity and Polymorphism of Functionally Specialized Blood Proteins in Migratory Fish: The Case Study of the North Caspian Population of the Russian Sturgeon during Sea and River Periods of Life. 1. Albumins

A comparative study of the levels of heterogeneity and polymorphism of albumins, the most important functionally specialized blood proteins, has been carried out. The albumin system of Russian sturgeon undergoes distinct changes while the fish change habitat during the spawning migration from sea to river. They are expressed as an increased level of heterogeneity, an increased content of serum albumins in fish during the river period of life as compared to the sea period, and an increased share of a slow component of albuminograms. These changes suggest a significant role of the blood albumin system in adaptation of the Caspian sturgeon migratory species to fresh water life conditions.     

Age-related variability of serum protein antigens in sturgeons (Acipenseridae, Acipenseriformes)

We studied the variability of antigenic properties of proteins in two sturgeon species at different stages of postembryonic development. The deepest changes occurred in individual components of albumins and beta-globulins (transferrins) and were mostly related to an increased proportion of the protein accounting for these antigens. Transformation of the main component of albumins A1 into adult antigens was completed in 5-month old fry. The main component of beta-globulins A (component of transferrins) appeared in the blood flow much later than other proteins and could retain the fry features until the age of 3-4 years. Other antigens belonging to alpha1- and alpha2-globulins and the second component of transferrins were more stable and did not undergo substantial changes. The direction of ontogenetic variability of serum antigens in sturgeon fry did not depend on the habitat of adult fish in fresh or sea water.     

Characterization of Albumins fromLupinus luteus L. Cotyledons

The cotyledons of yellow lupin seeds contain 15.4 mg of the albumins, water soluble proteins, per 100 mg of dry matter. These proteins were resolved by SDS gel electrophoresis into 13 polypeptides with relative molecular mass, Mr, between 16 000 and 84 000. Isoelectrofocusing analyses of the albumins showed that they were composed of six components with isoelectric points between 4.1 and 5.5. These results have indicated that the albumins are acid proteins and exhibited molecular and charge heterogeneity.     

Age-Dependent Variation of Serum Protein Antigens in Sturgeons (Acipenseridae,Acipenseriformes)

We studied the variation of antigenic properties of proteins in two sturgeon species at different stages of postembryonic development. The deepest changes occurred in individual components of albumins and -globulins (transferrins) and were mostly related to an increased proportion of the protein accounting for these antigens. Transformation of the main component of albumins A1 into adult antigens was completed in 5-month old juveniles. The main component of -globulins A (component of transferrins) appeared in the blood flow much later than other proteins and could retain the juvenile features until the age of 3–4 years. Other antigens belonging to ₁- and ₂-globulins and the second component of transferrins were more stable and did not undergo substantial changes. The direction of ontogenetic variation of serum antigens in sturgeon juveniles did not depend on the habitat of adult fish in fresh or sea water.     

A note on the fecundity of pallid sturgeon

Fecundity was estimated for two pallid sturgeon, Scaphirhynchus albus, from the Lower Mississippi River (LMR). Sturgeon measured 827 and 886 mm fork length, weighed <3.2 kilograms and had 16 237 and 17 810 ova per kilogram of body mass (based on mean ovum density) with total fecundity estimates ranging from 43 357 to 58 913. Ovum size in the two sturgeon was 2.5–3.0 mm diameter. Pectoral fin ray sections showed distinct annuli and spawning bands. The smaller sturgeon was aged at 10 and the larger fish at 12 years. Spawning bands were present and suggest the smaller sturgeon first spawned at age 8 and the larger fish at age 9. When compared to published results, the data representing specimens from more northern populations suggest that pallid sturgeon from the LMR mature at comparable ages but at smaller sizes and have greater gonadal mass. These features and greater fecundity per mass unit (number of eggs per kilogram body weight) suggest that LMR pallid sturgeon populations exhibit greater potential for population growth and recovery than other populations with slower growth and lower fecundity.     

New strategies for brood stock management of sturgeon in the Sea of Azov basin in response to changes in patterns of spawning migration

The sturgeon stock in the Sea of Azov is presently maintained by artificial reproduction allowing for the harvest of sturgeons in this basin. Shortened spawning migrations in regulated rivers and long-term artificial selection of the ripest fish at the hatcheries have resulted in high functional maturity of sturgeon spawners in the sea near the river mouth. The traditional technology of artificial sturgeon reproduction is non-effective under modern ecological conditions. The use of ecological and hormonal methods to control seasonal propagation of stellate sturgeon, A.stellatus , Russian sturgeon, A.gueldenstaedti , beluga, Huso huso , and sterlet, A.ruthenus , make it possible to stagger the sexual cycle of breeders from the various biological groups for early (5 months) or later (6 months) spawning. Use of these methods preserves the natural heterogeneity of sturgeon populations maintained through stocking, and also allows for a considerable increase in the quantity of fry reared at the same production base.
The use of the synthetic analogue of mammalian gonadotropin-releasing hormone (Gn-RHa) is better for inducing maturity of the ripest brood fishes, especially for stellate sturgeon, in comparison with the traditional pituitary injections. The schedule of injections was determined for the various species and seasons of spawning.
Short-term rearing of larvae in ponds (no more than 15 days) and subsequent adaptation to sea water in brackish lagoons (1–5 %o) allows for repeated use of nursery ponds in sturgeon hatcheries and a considerable increase in the quantity and survival of sturgeon juveniles in the sea.
The restoration of the sturgeon population structure in the Sea of Azov is not possible without developing a genetic collection of the fish under a controlled culture regime.     

Polymorphism of the myogens locus in some representatives of sturgeons (Acipenseridae)

This study investigates muscle proteins in Russian (Acipenser gueldensiaedtii Brandt), Siberian (Acipenser baerii Brandt), and Amur (Acipenser schrenckii Brandt) sturgeons using the method of polyacrylamide gel electrophoresis. A group of fractions, which is presumably the product of a polyallelic duplicated MY-1* locus, is recorded among the general pool of proteins. Data from densitometric analysis makes it possible to put forward a hypothesis about an eight-gene determination of this system of proteins. The highest heterogeneity and polymorphism was revealed in the Russian sturgeon. Analysis of the genotype distribution in samples of the studied species allowed us to make an assumption that the genetic structure of spawning sturgeon stocks is significantly metamorphosed under the influence of anthropogenic and natural factors.     

The status and distribution of lake sturgeon, Acipenser fulvescens, in the Canadian provinces of Manitoba, Ontario and Quebec: a genetic perspective

Genetic analysis of mitochondrial DNA sequence variation indicates that most of a sample of 396 lake sturgeon, Acipenser fulvescens, from the northern part of their range belonged to either one of two haplotypes. The vast majority of fish from the Great Lakes/St. Lawrence and Mississippi drainages were of a single haplotype while those from the Hudson/James Bay were composed of both haplotypes. This haplotypic distribution suggests that fish from one refugium (possibly Missourian) recolonized the Hudson-James Bay drainage while those from a second (possibly Mississippian) recolonized the Laurentian Great Lakes and St. Lawrence River. Lake sturgeon still inhabit much of their native postglacial distribution in Manitoba, Ontario and Quebec. However, the stresses of commercial overexploitation and habitat alteration, usually through hydroelectric dam construction and operation, have either singly or in tandem brought about the reduction, if not extirpation, of some populations within the range. The largest zone of extirpation and population reduction has occurred in the Lake Winnipeg drainage area, which covers more than one-third of Manitoba. Other areas where populations have been reduced to remnant levels, if not extirpated, include the lower Laurentian Great Lakes of Lake Ontario and Lake Erie. In northern Ontario, lake sturgeon populations whose riverine habitats have been fragmented by two or more dams are substantially reduced from their former levels. In Quebec, more attention has been paid to limiting the exploitive stresses on lake sturgeon populations. Combination of the genetic and status data suggests that both northern and southern populations of lake sturgeon (possibly from two glacial refugia) have been impacted severely from anthropogenic influences.     

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.     

Discrete phenotypical variability of electrophoretical specters of one of system of myogens in the Acipenseridae family

Analysis of electrophoregrams of water-soluble proteins of white skeletal muscle of 10 species belonging to three genera of the Acipenseridae family has allowed revealing an earlier not described system that can serve as a biochemical marker. This protein system represents a set of five fractions (A, B, C, D, E). A part of the studied species is monomorphous for one of these fractions (sterlet, giant sturgeon, Caspian sturgeon, and Amu-Darya shovelnose had phenotype A, barbel sturgeon—phenotype C, Amur sturgeon—phenotype D). In all individuals of the Amur sturgeon the identical three-component specter (phenotype ACE) was revealed. In the Sakhalin, Siberian, and Russian sturgeons a clearly expressed polymorphism was observed. Among the studied species the Russian sturgeon was characterized by the highest degree of heterogeneity and polymorphism. In the studied sample of this species all five protein variants are found, whereas in the rest of them—only a part of this set.     

Distribution of mitochondrial DNA variation in lake sturgeon (Adpenser fulvescens) from the Moose River basin, Ontario, Canada

We analysed mitochondrial DNA (mtDNA) variation of lake sturgeon ( Adpenser fulvescens ) from the Moose River basin. Our objective was to address various proximate and ultimate factors which may influence the distribution of lake sturgeon mtDNA haplotype lineages in this watershed. The lake sturgeon sampled were characterized by only two mtDNA hapiotypes based on a restriction fragment length polymorphism analysis with 40 restriction endonucleases and direct sequencing of 275 nucleotides in the mtDNA control region. We detected no heterogeneity in the mtDNA haplotype frequencies of lake sturgeon captured from different sites within rivers including those separated by major hydroelectric installations. However, lake sturgeon from one tributary had significantly different haplotype frequencies than those from other tributaries suggesting that they composed a discrete genetic stock. These results suggest that gene flow among most sites is significant and is an important factor affecting the distribution of mtDNA variation in this species. The genetic structuring and diversity are discussed in relation to lake sturgeon management and conservation.     

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.     

Peculiarities of secondary structure of serum albumin of some representatives of the animal kingdom

Methods of infrared (IR) spectroscopy and circular dichroism (CD) are suitable techniques for detection of proteins structural changes. These methods were used for determinating peculiarities of the secondary structure of serum albumins in some representatives of two classes of reptiles: Horsfield's tortoise (Testudo horsfieldi), water snake (Natrix tessellata) and grass snake (Natrix natrix) and birds: domestic goose (Anser anser), domestic chicken (Gallus domesticus), domestic duck (Anas platyrhyncha) and dove colored (Columba livia). An analysis of IR spectra and spectra obtained by the method of CD of serum albumins of both classes representatives revealed that beta-folding structure and alpha-helical sections that form the alpha-conformation play an important role in conformational structure formation of polypeptide chain and also disordered sites of molecules of these proteins. It was observed that certain redistribution depending on animals species exists, in the formation of secondary structure of serum albumins of the investigated representatives of reptiles and birds classes between the content of beta-folding structure, alpha-helical sections and disordered sites in molecules of these proteins.     

Synergistic Enhancement of the Antifungal Activity of Wheat and Barley Thionins by Radish and Oilseed Rape 2S Albumins and by Barley Trypsin Inhibitors

Although thionins and 2S albumins are generally considered as storage proteins, both classes of seed proteins are known to inhibit the growth of pathogenic fungi. We have now found that the wheat (Triticum aestivum L.) or barley (Hordeum vulgare L.) thionin concentration required for 50% inhibition of fungal growth is lowered 2- to 73-fold when combined with 2S albumins (at sub- or noninhibitory concentrations) from radish (Raphanus sativus L.) or oilseed rape (Brassica napus L.). Furthermore, the thionin antifungal activity is synergistically enhanced (2- to 33-fold) by either the small subunit or the large subunit of the radish 2S albumins. Three other 2S albumin-like proteins, the barley trypsin inhibitor and two barley Bowman-Birk-type trypsin inhibitor isoforms, also act synergistically with the thionins (2- to 55-fold). The synergistic activity of thionins combined with 2S albumins is restricted to filamentous fungi and to some Gram-positive bacteria, whereas Gram-negative bacteria, yeast, cultured human cells, and erythrocytes do not show an increased sensitivity to thionin/albumin combinations (relative to the sensitivity to the thionins alone). Scanning electron microscopy and measurement of K+ leakage from fungal hyphae revealed that 2S albumins have the same mode of action as thionins, namely the permeabilization of the hyphal plasmalemma. Moreover, 2S albumins and thionins act synergistically in their ability to permeabilize fungal membranes.     

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

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

Microsatellite multiplex assay for the analysis of Atlantic sturgeon populations

We have developed a multiplex assay covering 16 microsatellite loci, amplified in four polymerase chain reaction (PCR) assays, and loaded on the ABI DNA Analyzer in two separate panels. The assay was tested on 603 individuals originating from wild populations and hatchery stocks of Atlantic sturgeon. The assay was also tested on 12 individuals of European sturgeon and appeared to be almost equally useful. The multiplex assay designed in this study can be successfully applied in studies requiring high genetic resolution, such as relatedness analysis, selective breeding programs, and stock identification of Atlantic 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.     

Rapid HPLC analysis of retinoids and dehydroretinoids stored in fish liver: comparison of two lake sturgeon populations

All major retinoids (vitamin A) stored in the liver of lake sturgeon from two Québec lakes were analysed by high-performance liquid chromatography (HPLC). Four HPLC peaks represented approximately 90% of all liver retinoids. Their relative proportion was remarkably consistent in the two sturgeon populations with dehydroretinyl palmitate/oleate accounting for the largest proportion (50–60%). Most retinoids demonstrated significant positive correlations with age, but only in the Lake Berthelot sturgeon. The concentrations of all major retinoids were significantly ( P <0·0001) lower, as much as 40 times lower, in the Lake Saint-Louis population. Possible explanations for the extremely low liver stores in the Lake Saint-Louis sturgeon are discussed.     

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.     

Bisalbuminemia in an amphibian

Improved electrophoretic resolution revealed two albumin-like proteins in Taricha granulosa plasma (bisalbuminemia). The Taricha proteins were compared to mammalian, avian and reptilian serum albumins regarding molecular weight, amino acid composition, isoelectric character, solubility and the binding of hemin and dyes. The results indicate that although the two Taricha proteins have demonstrated hemoglobin-binding ability, they possess traits that characterize them to be true serum albumins.