Results of cloning X69R, A179L, E248R, I215L and DP96R genes of ASF virus Krasnodar 07/17 isolate and analysis of their nucleotide sequences are presented. Obtained clones were added to the previously constructed clone library comprising clones of 8 genes of Krasnodar 06/12 isolate. Clones containing X69R, A179L, E248R, I215L and DP96R genes of ASF virus Krasnodar 07/17 isolate will be used for recombinant protein obtaining and testing for their effect on in vitro virus reproduction and their role in the virus infectivity, level of clinical manifestations and virulence. Prokaryotic vector, pJET1.2/ blunt, was used. Thus, the clone library available at the FGBI “ARRIAH” Reference Laboratory for African swine fever was supplemented by pJET1.2-X69R, pJET1.2-A179L, pJET1.2-E248R, pJET1.2-I215L and pJET1.2-DP96R plasmid constructions containing 5 genes of ASF virus Krasnodar 07/17 isolate. Proportion of cloned virus genes was 3.01% of Krasnodar 07/17 isolate genome, hence, total amount of the clone library has reached 7.82%.

Tests of inactivated emulsion anti-Aujezsky’s disease, porcine reproductive and respiratory syndrome (PRRS) vaccine based on marker Aujezsky’s disease virus strain, PRRS virus strain and inactivated emulsion anti-Aujezsky’s disease and porcine parvovirus infection vaccine based on marker Aujezsky’s disease virus strain and porcine parvovirus strain developed by the FGBI “ARRIAH” for their antigenicity are described in the paper. It is demonstrated that the said vaccines can be used successfully for PRRS and porcine parvovirus infection and Aujezsky’s disease prevention. They are becoming of special significance when used in programs on Aujeszky’s disease control and eradication. The use of the vaccine based on marker Aujeszky’s disease virus strain allows differentiation between the animals that have antibodies to the marker vaccine strain (in the absence of antibodies to the glycoprotein gE in 100% of cases) from the animals with antibodies to field strains of the Aujeszky’s disease virus in their sera. A comprehensive assessment of the use of associated vaccines developed in the FGBI “ARRIAH” (against Aujeszky’s disease based on the marker strain, reproductive and respiratory syndrome, parvovirus infection), in the framework of anti-epidemic measures on a pig farm of the Russian Federation showed their safety, high antigenicity, immunogenicity.

Results of retrospective analysis of African swine fever epidemic situation in the Russian Federation including analysis of the epidemic chronology indicating significant events of the disease spread across the country territory are presented. Common ASF epidemic trend and rate in 2007–2017 in Eurasia territory based on number of infected countries (1.273 ± 1.272 countries/year) and in Russian Federation Subjects (4.5 ± 2.3 Subjects/year) are shown. The paper also addresses some peculiarities of the epidemic process depending on the season of a year. Possible social and biological factors contributing to the epidemic spread are examined. Necessity of further search for probable biological vector and examination of social factors that could contribute to the disease maintenance in wild life and in domestic pigs in the Russian Federation and Eastern European countries are discussed.

Results of cloning X69R, A179L, E248R, I215L and DP96R genes of ASF virus Krasnodar 07/17 isolate and analysis of their nucleotide sequences are presented. Obtained clones were added to the previously constructed clone library comprising clones of 8 genes of Krasnodar 06/12 isolate. Clones containing X69R, A179L, E248R, I215L and DP96R genes of ASF virus Krasnodar 07/17 isolate will be used for recombinant protein obtaining and testing for their effect on in vitro virus reproduction and their role in the virus infectivity, level of clinical manifestations and virulence. Prokaryotic vector, pJET1.2/ blunt, was used. Thus, the clone library available at the FGBI “ARRIAH” Reference Laboratory for African swine fever was supplemented by pJET1.2-X69R, pJET1.2-A179L, pJET1.2-E248R, pJET1.2-I215L and pJET1.2-DP96R plasmid constructions containing 5 genes of ASF virus Krasnodar 07/17 isolate. Proportion of cloned virus genes was 3.01% of Krasnodar 07/17 isolate genome, hence, total amount of the clone library has reached 7.82%.

Method for determination of foot-and-mouth disease (FMD) virus 146S component concentration with real-time reverse transcription – polymerase chain reaction (rtRT-PCR) in vaccine raw materials is developed. Negative regression model of  С146S = (30.269 – Ct )/4.155 type allowing determination of FMDV 146S particle concentrations based on the amplification threshold cycle values (Ct ) is proposed. It has been experimentally proven that the quantity of the 146S component determined by the real-time RT-PCR method using developed regression model and contained in the inoculation dose of FMD vaccine confers protection to the vaccinated animals against generalized FMD of A, O, Asia-1 types. rtRT-PCR method is highly sensitive and allows rapid and highly reliable estimation of the 146S antigen concentration in FMD vaccine. The method for 146S particle quantity determination by real-time RT-PCR using the regression model is reliable and demonstrates high correlation (95.5–99.0%) with the complement fixation test results.

Predictive assessment of possible risks of FMD introduction from neighboring countries to the territory of eight RF Federal Districts was presented. The risk quantitative parameters were determined by experts, specialists in the field of FMD epidemiology. To implement the expert survey method most significant routes of infection introduction to the RF were determined. The experts performed FMD introduction risk assessment for each federal district and determined its score. As a result of statistical analysis the greatest probability was determined for the Far-Eastern Federal District. The North Caucasus and Siberian Federal Okrugs demonstrate lower probability. Basing on the obtained data the major routes of FMD introduction to the territory of the country were determined. Preventive vaccination of susceptible animal population is carried out in order to prevent FMD occurrence and spread in the zones at risk of its introduction. It is aimed at FMD outbreak prevention in the specified RF Subjects by inducing protective immunity in at least 81% of immunized cattle and at least 95% of immunized pigs.

Method for determination of foot-and-mouth disease (FMD) virus 146S component concentration with real-time reverse transcription – polymerase chain reaction (rtRT-PCR) in vaccine raw materials is developed. Negative regression model of  С_146S = (30.269 – C_t)/4.155 type allowing determination of FMDV 146S particle concentrations based on the amplification threshold cycle values (C_t) is proposed. It has been experimentally proven that the quantity of the 146S component determined by the real-time RT-PCR method using developed regression model and contained in the inoculation dose of FMD vaccine confers protection to the vaccinated animals against generalized FMD of A, O, Asia-1 types. rtRT-PCR method is highly sensitive and allows rapid and highly reliable estimation of  the 146S antigen concentration in FMD vaccine. The method for 146S particle quantity determination by real-time RT-PCR using the regression model is reliable and demonstrates high correlation (95.5–99.0%) with the complement fixation test results.

Studies have been carried on antigenic and protective activity of the experimental associated vaccine containing, in the field vaccination dose, antigens of the attenuated ARRIAH strain of the sheep pox virus and ARRIAH 2003 strain of the goat pox virus in the ratio of 4.24 and 4.24 lg TCD₅₀/cm³, 4.18 and 4.37 lg TCD₅₀/cm³ and 4.37 and 4.18 lg TCD₅₀/cm³ respectively, as well as monovalent vaccines with the infectious activity of these strains with a titer of 4.5 lg TCD₅₀/cm³. Administration of the associated virus vaccine did not adversely affect the physiological state of the animals and was accompanied by the formation of viral neutralizing antibodies in protective titres that did not differ from antibody titers in the blood of sheep and goats during monovalent immunization. High levels of virus neutralizing antibodies in the range of (3.50 ± 0.50)–(4.05 ± 0.22) and (3.58 ± 0.08)–(4.00 ± 0.29) log₂ respectively were determined in the blood serum of all sheep and goats vaccinated with associated vaccines. The antigenic activity of the associated vaccines for goats and sheep is almost identical. There were no negative effects on the antigenic activity of different ratios of attenuated strains of the sheep pox and goat pox virus in the inoculation dose. All sheep immunized with the monovalent vaccine against sheep and goat pox and associated virus vaccines, resisted the disease when challenged with a virulent sheep pox virus “Afghanistan” strain. Three sheep vaccinated with a monovalent vaccine against goat pox demonstrated a large local reaction, one sheep was diagnosed with generalized form of sheep pox.

The raw material for blood protein hydrolysate preparation is whole animal blood, its clots and other serum production wastes. The dependence of amino acid composition of blood protein hydrolysate on the season of the raw material preparation was studied. The research lasted three years. It was demonstrated that the amino acid composition changed depending on the season. The peak, as a rule, was during summer months when their amount increased by 1.2–2.3 times and during autumn and winter it went down by 1.2–1.4 times (the difference is considerable, р < 0,05). The peak of glutamic and asparagine acid growth was in November when their amount was 1.4 times higher then during the previous months (р < 0,01). The increase of alanine, asparagine, valine, lysine, methionine, histidine, proline, tyrosine, threonine, and phenylalanine by 1.3–1.8 times was observed in March (the difference is considerable, р < 0,05). The amount of histidine, glycine, leucine, serine, and tryptophane in the beginning of spring was at the same level and the amount of arginine, asparagine, isoleucine in March decreased by 1.2–1.6 times (the difference is considerable, р < 0,01). So, it was determined that the dynamics of BPH amino acid composition was directly associated with the seasonal dynamics of physiological and biochemical cattle blood values. It was noted that in case of considerable change in absolute amino acid parameters their relative amount, in general, remained constant.

The Amur Oblast belongs to biogeochemical province with low contents of iodine in the soil, water and air, which leads to the thyroid gland disorders in animals, weakened immunity system and, consequently, to the increased sensitivity to diseases. A new iodinecontaining preparation “Vangtseyod” has been developed basing on natural zeolites from Vanga deposits to prevent iodine deficit in young cattle. The distinctive feature of the Vanga deposit zeolite in comparison with other deposits of the Amur Oblast and the Far East is that it contains more effective clinoptilolite and heulandite minerals, characterized by the low contents of harmful compounds. Studies of the preparation safety performed on white mice showed that “Vangtseyod” does not have acute toxicity. It’s inclusion into mice diet at 1.0 and 2.0 g/kg dose during two months did not induce any signs of chronic intoxication as 100% mice of experimental group survived in comparison with the control group (57%). Feeding mice with the preparation at 1.0 g/kg dose facilitated improved protein and carbohydrate metabolism.

It is known that predatory Hyphomycetes fungi play an important role in measures taken for helminthic infestation prevention. There is little evidence on properties of compounds contained in predatory fungi and their effect on animals. Results of tests of "Vetom 21.77", new microbiological preparation based on Duddingtonia flagrans, for its effect on serum biochemical pattern of laboratory mice are presented. The preparation was administered to the animals at the dose of 2, 5, 50 and 300 µl/kg of body weight. The animals were examined for tested parameters prior and 2 and 7 days after daily administration of the preparation. The parameters remained within the normal physiological limits in all test group as well as in control group not given the preparation. The test results indicated safety of the said microbiological preparation. Significant increase in total protein and albumin contents was recorded in mice given the maximum dose of the preparation as compared with those of control group.

New method of glycogen determination in bee organ tissues, considerably different from the existing ones, was suggested. Ghoreishi’s colorimetric method used today is time-, labor- and cost-consuming as the major reagent – orcin – is quite expensive. Comparison of cost-effectiveness of the two methods demonstrated that the proposed technique makes it possible to reduce expenses by using a more available and less expensive resorcin instead of orcin and reducing total test time from 4 to 3 hours 5 minutes. Glycogen contents determination using the updated method was performed in bees of four breeds: Italian-Carpathian, Carpathian, Oka, gray Caucasus mountain honeybees. It was determined that the Oka honeybees demonstrated the highest glycogen level. It is known that the higher is glycogen contents in organs and tissues the better is energy metabolism in insects. Consequently, honeybees of Oka breeds have higher resistance to unfavorable weather conditions and can produce progeny with a higher level of immunity. So, the specified method of glycogen determination in bee organ tissues increases the accuracy of diagnosis and plays a very important role in determining the level of energy metabolism in insects and will be useful for apiculture.

Age-related dynamics of hematological and functional parameters of Pekin duck blood during administration of organic selenium preparation DAFS-25k (at a 1.3 mg/kg dose) was presented. In order to study morphological parameters from the age of one day old to 120 days old blood was collected from axillary vein of birds prior to feeding each 15 days in the morning. To assess hematological parameters RBC and WBC count was performed, hemoglobin contents and hematocrit value were determined and color index of blood was calculated. The experiment demonstrated gradual increase in these parameters in the experimental and control groups. Birds from the experimental group demonstrated higher parameters than those of the control group. It was determined that hemoglobin contents was 9.86% higher in the experimental group in comparison with the control group due to selenium administration. The blood color index in this group was also higher (р ≤ 0.05). Biochemical blood analysis included the following parameters: total protein, albumin, uric acid, glucose, calcium, phosphorus, ALT and AST transaminase. The test results showed that during all age periods total protein and albumin contents in ducks of the experimental group exceeded those of the control group. The selenium-containing preparation had a positive influence on the carbohydrate metabolism ensuring high level of metabolic processes in birds, contributed to optimal calcium to phosphorus ratio which positively influenced mineral metabolism and development of the locomotor system. Obtained AST and ALT values in both groups are not beyond reference values which indirectly confirms absence of toxic effect on ducks’ organism if selenium is administered at the specified doses. It was determined that DAFS-25k has not affected hematological and functional parameters of Pekin duck blood and it contributed to mineral metabolism correction, reduction of stress factor effect within critical periods of development and improvement of the excretory system function.