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In blood and tissue fluid of squamulose redeye, rock perches, starlet, and two species of electric rays, with aid of methods of proteomics, there were identified ependemines, vitellogenins, immunoglobulins, and albumin-like proteins (ALP). For all proteins except for ALP, identification was successful with use of MALDI mass-spectroscopy in the tandem regime. Reconstruction of fragments of amino acid sequences was performed with databases of fish with the sequenced genome and mammals. Problems at the ALP identification are connected with a high level of their structural diversity and the absence of similarity with mammalian proteins.

A wide range of differences of cations concentrations (Na, K, Ca, Mg) in hemolymph, blood serum of multicellular animals and humans was revealed. It was established that constancy of K/Na ratio 0.034 +/- 0.001 is strictly maintained in these organisms. Except for some freshwater and marine organisms, Mg/Na ratio is sustained on the same level in the wide range of concentrations of these ions in serum of vertebrates and hemolymph of invertebrates. Ca/Na ratio is higher in hemolymph of freshwater mollusks. The question about possible reasons of physiological differences of Ca/Na and Mg/Na rations compared to constancy of K/Na ratio is discussed.

To investigate interaction of atherogenic low-density lipoproteins (LDL) with erythrocytic membrane, the content of lipid components in blood cells and serum LDL was studied in human in norm (donors) and in 12 species of vertebrates (the mammals non-predisposed to atherosclerosis - birds and fish). Lipid composition of blood cells and LDL was analyzed also in patients with pathologies: ischemic heart disease (IHD), bronchial asthma (BA), and chronic obstructive bronchitis (COB), and in 2 species of mammals predisposed to atherosclerosis, in whose blood LDL predominates. The content of lipids in cells and LDL of the studied vertebrates has been found to depend on their taxonomy and the clear trends both to an increase of the cholesterol content and to a decrease if the phosphatidylcholine level in patients, particu- larly with IHD, and on a rise of the ratio of the content of the more saturated sphingomyelin and cholesterol to the less saturated phosphatidylcholine from the lower to the higher organisms, including humans (donors). The highest levels of free cholesterol in blood cells, of total cholesterol in LDL, and of ration of the cholesterol/phosphatidylcholine content have been revealed in patients, especially with 1HB, and in the mammals predisposed to atherosclerosis, i. e., in representatives with predominance of blood LDL, unlike donors and the mammals resistant to atherosclerosis. The highest parameters of lipid components were determined in cells and LDL inhuman with IHD. The lipid LDL composition affects directly the composition and ratio of lipids in blood cells.

Influence of temperature in the range of 1-15 degrees C on oxygen binding properties of blood of thermophilic--golden mullet (Liza aurata), anchovy (Engraulis encrasicolus), and cold-tolerant--sardelle (Clupeonella cultriventris) fishes has been investigated under experimental conditions. Heat dependence of oxygenation reaction in thermophilic fish blood at temperature below 10 degrees C considerably increases, which is evidenced by high deltaH values. That is accompanied by a substantial increase of blood oxygen affinity and complicates blood deoxygenation at the tissue level. This reaction is apparently determined by the change of hemoglobin interaction with intraerythrocyte medium. The concentration of NTP in erythrocytes increases, that partially compensates negative changes of blood oxygen affinity (parameter P50 is raised) under long-term maintenance of fishes at 5 degrees C. However this reaction is not observed at low temperatures (1-2 degrees C).

Blood proteins of true sturgeons of three genera Acipenser, Huso, and Pseudocaphirhynchus preserving unidirectional variability in ontogenesis regardless of peculiarities of the fish ecology can indicate unity of origin of the current species. Age-related protein transformations have heterochronous character. The slowest ontogenetic development lasting 4-6 years is peculiar to one of antigens of beta-globulins. Analysis of variability of beta-globulins in the age diapason beginning from the 4-month-old fry and including adult spawned individuals indicates different functional significance of their individual components. The beta A-globulin absent at early postembryonic stages becomes the second, after albumin, quantitatively predominant antigen in the adult fish blood. The amount of beta B-globulin, like antigens of alpha 1- and alpha 2-globulins preserving relative ontogenetic stability, can decrease significantly in freshwater sturgeons due to their preparation for spawning.

The effect of photoperiod on the growth of juvenile Siberian sturgeon Acipenser baerii has been studied in aquarium experiments. The maximum growth rate of juvenile Siberian sturgeon was observed under 12, 16, and 24 h photoperiods. This was accompanied by high respiratory rate and total food consumption and conversion. Rearing in the dark significantly decreased these indices. The hematological indices were within the normal range under optimal photoperiodic conditions. Pronounced neutrophilia and leukopenia were observed in the absence of light.

We studied blood serum albumins in European (Acipencer ruthensis L.) and Siberian (A. ruthensis marsiglii Brandt) sterlet using disk electrophoresis in polyacrylamide gel. The albumins were shown to be controlled by three codominant alleles of a single locus (ALB*a, b, c). In European sterlet, all three theoretically possible genotypes were described, one of which (ALB*c/c) occurred extremely rarely (one individual). Siberian sterlet was found to be monomorphic for albumins: all fish examined had the ALB*a/a genotype. There was no correlation between albumin patterns and fish fatness. In a number of samples from the Volga River basin, spatial and temporal differentiation was found and analyzed. The results suggest that construction of hydroelectric plants may provoke massive and prolonged starlet migrations.

The subject of the case study was blood of 8 fish species. It was examined within the ABO system. 68 tests of stains of fish blood mixed with human blood were made. It was shown as possible to differentiate between human blood and fish blood in mixed stains.

It was shown that the phenotypes of haptoglobin (Hg) can not be detected in stains of dried blood of salmon, roach and bream. The results of experimental research of blood of the above fish species are described according to the Cm system. It was proven as possible to identify the human blood in stains with admixtures of blood of the above fish by using the Hp and Cm systems.

Blood serum cortisol, testosterone, and 11-ketotestosterone levels were determined by ELISA methods in female Russian sturgeon after hormonal stimulation for induction of ovulation. All females were divided into 5 groups depending on their initial reproductive status and response to hormonal stimulation; 1) normal ovulation, 2) ovulation followed by release of non-viable eggs, 3) normal state of ovary without ovulation, 4) lack of ovulation, early stage of oocyte resorption, 5) lack of ovulation, advanced stage of oocyte resorption. Cortisol levels in all five groups did not differed significantly. Testosterone levels were low (15-24 ng/ml) in females after ovulation and in females without response to hormonal stimulation because of early stage of oocyte resorption. Non-matured females with normal state of oocytes had significantly higher testosterone levels--82 +/- 16 ng/ml. Non-matured females with advanced stage of resorption were subdivided into 2 subgroups--with low (approximately 7 ng/ml) and high (> 100 ng/ml) levels of serum testosterone. 11-ketotestosterone levels were similar in all investigated groups of sturgeon.

Serum steroids profiles were determined at different stages of the migratory cycle of Russian sturgeon. In the sea period, before gonads maturation cortisol and sex steroids levels were comparatively low. Elevation of cortisol and testosterone concentrations occurred at the stage of preparation to migration. Significant increase of cortisol and testosterone levels happened at the beginning of the river period of anadromous migration in spring both in the spring forms near to maturity in this period and in the winter forms which were far from maturity. By exception of the river period of anadromous migration of the winter form and sexual cycle completion in the fish farm in the mouth of river (10-11 months), serum cortisol and testosterone concentrations dropped sharply. The data obtained as well as the data on other diadromous fishes indicate a possible role of steroids, especially cortisol and testosterone, in metabolism and migration regulation in sturgeons.

A comparative analysis of two types of cryoprotectants, antifreeze glycoproteins from Atlantic cod (Gadus morhua) and carbohydrates from hemolymph of amphypoid crustacean (Gammarus lacustris) has been performed. Both glycoprotein and carbohydrate antifreezes effectively decreased the freezing point of water solutions and diminished the size of ice crystals formed. Noncolligative and colligative mechanisms of action are characteristic correspondingly for glycoproteins and carbohydrates.

The levels of chromosome mutations in hemopoietic cells of various fish from water bodies near the Noril'sk mining and smelting complex were estimated by the micronucleus test. In some cases very high frequencies of erythrocytes with micronuclei were observed in fish inhabiting highly polluted water bodies. On the whole, the number of cells with chromosome aberrations was within the limits of spontaneous mutagenesis. Marked species-specific, seasonal and local differences were found in the frequency of erythrocytes with micronuclei. The problems related to the influence of chemical pollutants on fish are discussed.

Using fluorescence spectroscopy species peculiarity of hydrophobic and microviscosity degree in central and peripheral molecular areas of the main electrophoretic haemoglobin fractions in representatives of different vertebrate classes gas been studied. It is established that central areas of haemoglobin molecule are characterised by twice lesser variability of size and hydrophobic degree and 2.4 times greater variability of package strength as compared with peripheral areas of protein globule.

The cryoprotective properties of antifreeze glycoproteins (AFGPs) isolated from White Sea cod (Gadus morhua) blood were analyzed. Improved preservation of morphological characteristics was observed in two-cell mouse embryos frozen in the presence of AFGP but without other cryoprotectors. However, subsequent development after thawing was achieved only with embryos frozen in a medium containing both AFGP and dimethyl sulfoxide. The maximum increase in the number of surviving embryos (by a factor of approximately two, as compared with the control) was observed at AFGP and DMSO concentrations of 20 mg/ml and 25%, respectively.

Hypoxia effect on the nuclear of the Scorpaena porcus (L.) in vivo was studied. It was shown, that existence of the fishes in environmental with low oxygen concentration-1.3-1.4 mg.1-1 (15% initial saturation) resulted in reducing in activities of Na+, K(+)-ATPase, hexokinase and glucose-6-phosphate dehydrogenase in the erythrocyte for 50.0 (p < 0.001), 26.5% (p < 0.01) and 53.7% (p < 0.05) accordingly. ATP concentration in cells and membrane gradient of Na+, K+ concentrations between blood serum and intracellular environment did not change. A conclusion was made about a decrease of cells membrane penetration and oppression of intracellular metabolism. These changes proceeded on a background of the blood serum dehydration and decrease of the mean volume of erythrocytes. The part of aldosteron and vasopressin in membrane penetration of nuclear erythrocytes is discussed.

Using isoelectrofocusing within pH gradient 6-8, studies have been made on microheterogeneity of haemoglobins from various vertebrates. Phylogenetic evaluation of oxygen affinity of the main and minor electrophoretically pure haemoglobin fractions is given. It was shown that oxygen affinity of the haemoglobins is related to the ratio between dicarboxylic and basic amino acids.

In reaction of hydrolysis of choline and thiocholine esters of carbonic acids at 25 degrees C, cholinesterase activity of the blood serum from the fish A. ballerus has been studied by modified Ellman's method and potentiometric titration method. The activity is maximal in pH region 7.5-9.0 and is not inhibited by high concentration of substrates. Michaelis constants and maximal rates for the enzyme reactions were determined. Butyrylcholine and butyrylthiocholine were hydrolyzed with the highest rates by the serum. Some of the organophosphorus inhibitors (diisopropylfluorphosphate and DDVF) inhibit cholinesterase activity of the blood serum significantly faster, whereas some of the carbamates (aminostygmin, eserine, etc.) inhibit it significantly slower than typical butyrylcholinesterase from horse blood serum and typical acetylcholinesterase of human erythrocytes. Besides, with respect to the sensitivity to inhibitors and some other properties, fish blood serum cholinesterase differs from other known cholinesterases.

The method of analysis of low molecular weight fragments of high copied repeats of DNA hydrolysed by restriction nuclease is presented. The P labeled fragments (by means of DNA polymerase I) are electrophoresed in nondenaturing PAAG and radio-autographed. The specific band patterns are observed in region between approximately 20 and 300 bp. When studying some lizard and fish species DNA's it was shown that the patterns observed have species and genus specificity but not individual. The approach supposed can be applied to investigation of interspecies relationships, some evolutionary problems and to the studying of questions concerning the role of DNA repeats in evolution. The method is simple and comparatively cheap and may be called as "taxonomic DNA fingerprint" or "DNA taxonoprint" method.

Electrophoretic method of simultaneous determination of blood presence and its human (primates), bird and fish origin was developed. Method is based on detection of diagnostical characteristics possessing specific electrophoretic mobility and doesn't require use of precipitating sera. Diagnostical feature of human (primates) blood is methaemalbumin detectable in anodic part of electrophoregram and not detectable within the given test conditions in bloodstains of other animals. In bird and fish blood haemoglobin is determined in cathodic part of electrophoregram which differs from mammals blood by its electrophoretic mobility.

Corticosteroid content of the blood has been measured during upstream and downstream migrations as well as during sea period of life of the sturgeon A. galdenstädti. It was shown that during anadromous migration before spawing, the level of corticosteroids in the blood increases, the increases beginning already during the sea period. Corticosteroid content of the blood decreases after spawning. The results obtained confirm the histophysiological data.

Studies have been made on fractional composition of hemoglobin in the Russian Acipenser güldenstädti, Siberian A. baeri and Amur A. shrencki sturgeons by polyacrylamide gel disc-electrophoresis. High electrophoretic heterogeneity (6--7 components) of hemoglobin in each of the species was found together with a distinct species specificity of hemoglobinograms. The observed differences in fractional composition of hemoglobin are of an adaptive character which makes it possible to consider the Amur sturgeon as a perspective object for acclimatization in the seas of the Southern region of the USSR with lowered salinity.

The absorption spectra from met-haemoglobin from lamprey Lampetra fluviatilis and its complexes with F-, CN-, N3- and imidazole were studied in non-equilibrium states arising in low-temperature (77K) reduction of water-glycerol of oxidized forms of protein with thermolyzed electrons. The Soret band was shown to be not split in low-temperature absorption spectra of reduced met-haemoglobin and its complexes unlike that of the equilibrium deoxyhaemoglobin. It was concluded that met-haemoglobin has a higher symmetry of the active centre as compared to deoxyhaemoglobin.

A radioimmunoassay was to determine the content of estrogens, progestins and androgens in the blood plasma of female and male big-mouthed buffalo, silver carp and canal sheatfish before reproduction and in the first two species during reproduction as well. It was shown that sexual hormones of all 3 groups were present in the blood of males and females of the species under study, the androgen concentration being significantly higher than the estrogen and progestin content in species of both sexes. Sexual dimorphism in the estrogen and androgen content in the blood of the fishes before reproduction was revealed. An exception was a silver carp with the absence of sex differences in the androgen content. There were no significant differences in the initial progestin levels in the species of both sexes. Sex differences in the time-course of gonadal hormones during reproduction were revealed: an increase in the level of steroids of all 3 groups was observed during ovulation in females, that of androgens and progestins only in females of the studied species excluding silver carp. No increase in the concentration of androgens was observed in male silver carps during reproduction. In spite of the general regularities of the time-course of sexual steroids in the reproduction of the studied species of fish significant specific differences were noted in the absolute values of sexual hormone concentrations.

Comparative studies on fractional composition of the blood serum proteins in two sympatric populations of the sturgeon A. stellatus (South-Caspian and North-Caspian) have been made by means of polyacrylamide gel disc-electrophoresis. Serum proteins are fractionated into 13-18 electrophoretic components, the heterogeneity of proteins being somewhat higher in the North-Caspian population than in the South-Caspian one. Most pronounced differences were found in the relative content of albumins and beta-globulins. Special interest is attracted to different heterogeneity of albumins and beta-globulins (transferrins) in the two populations of the Caspian sturgeon.

The results of studies of the quantitative values of nonspecific antihemagglutinins of influenza viruses in the sera of birds, laboratory, wild and domestic animals (altogether 27 species) are presented. The antiviral inhibitors characterized by a number of physicochemical properties (sensitivity to heating, KIO4, trypsin, rivanol, 2-mercaptoethanol) were divided into 3 groups, sera of sheep, goats and cattle making up a separate group with regard to their sensitivity to heating and treatment with KIO4. Studies using molecular screen chromatography demonstrated the nonspecific inhibitors present in bovine sera to be heterogenous both in type (thermolabile and thermostable) and in the molecular composition. Alongside with thermolabile inhibitors of macroglobulin nature, thermostabe 19S and 4S inhibitors were identified.

The influence of plasmin on blood coagulation in fishes has been studied. The plasmin administration in dose 250 conditional units has been shown to evoke hypercoagulation already on the 5th minute after injection. This defence reaction on the plasmin excess in blood flow was excluded by the preliminary administration of alpha- as well as beta-adrenoblocators. Subtle differentiation of adrenoreceptors is supposed to be absent in fishes, nevertheless they play a definite role in regulation of hemostasis system.

During adaptation of the skate Dasyatis pastinaca (L) to distilled sea water (7.1%) resulted in decreasing concentration of sodium, urea in blood serum, reduction in the chloride cells volume, their nuclei and increase of the apical part of the cell free from mitochondria (and occupied by the endoplasmic reticulum) are observed. Owing to uniformity of the morphometric changes of the chloride cells revealed in two types, during adaptation of the skate to the distilled sea water, as well as adaptation to its increased salinity, it is possible to conclude that in the gill of selachians there are chloride cells of one type with various functional activity. Adaptation of the skate to the distilled sea water is connected with removal of lipid inclusions out of the cells of the interrenal gland.

By means of polyacrylamide gel disc-electrophoresis studies have been made on a serum proteins of the kaluga Huso dauricus and Amur sturgeon Asipenser schrencki. High electrophoretic heterogeneity (18--23 components) of serum proteins of both species was demonstrated together with ecological peculiarities of fractional composition of these proteins as compared to those in Caspian sturgeons. The described high heterogeneity of serum proteins in the ganoid fishes is particularly interesting from the standpoint of evolution of fractional composition of proteins in vertebrates. Presumably, high heterogeneity of serum proteins in the ganoids (practically comparable to that in higher vertebrates and man) is necessary for successful work of the most important biochemical system of the blood at all stages of evolutionary development of vertebrates, being of rather ancient origin.

In all the animals studied (the lamprey Lampetra fluviatilis, teleosts Onchorhynchus nerka, O. kisutch, Limanda aspera, Salvelinus malma, frog Rana ridibunda, tortoise Agryonemys horsfieldi, hen, albino rat) and man, sodium and potassium were found in the blood serum in a free form, both before and after the addition of their salts in vitro. The initial level of active Ca varies within 20--42% of its total content in the serum. Significant interspecific differences were found with respect to the degree of Ca binding by albumin.

Comparative studies have been made on the kinetics of thermal denaturation of the blood plasma of various vertebrates (lampreys, teleosts, frogs, tortoises, pigeons, mice, rats, rabbits, cats, dogs, man) by measuring ionization equilibrium of protein solution at elevated temperature. It was demonstrated that during the initial stage of heat denaturation at 58 degrees the blood plasma of all the animals studied binds protons practically linearly. Among the animals studied, the highest protonoacceptor capacity exhibited the plasma of warm-blooded animals; it was lower in tortoises frogs and teleost fishes, being the lowest one in lampreys. Comparison of total electric charge of plasma proteins evaluated by potentiometric titration with data on thermal denturation of plasma revealed positive correlation between the charge and protonoacceptor properties of the plasma.