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Information × Registration Number 0216U005599, 0111U000466 , R & D reports Title To study the molecular genetic properties of enteroviruses suis patterns of variability and heterogeneity of the populations of enteroviral popup.stage_title Head Romanenko Volodimir Pilipovich, Registration Date 05-02-2016 Organization The Institute of veterinary medicine of Ukrainian Academy of Agrarian Sciences popup.description2 Objects of research: pig farms in different regions of Ukraine; pigs of all ages; samples of rectal swabs; serum pigs; Virus isolates; reference and epizootic strains of enterovirus swine plaque clones specific serum; horse serum, pigs, rabbits; cell culture CHEB Objective: To investigate the molecular and genetic properties of enteroviruses of pigs, the patterns of variability and heterogeneity of enteroviral populations Stage 1: To monitor the research and selection of isolates of enteroviruses of pigs in different regions of Ukraine, explore their cultural and biochemical properties. Stage 2: To study the molecular and genetic properties of selected isolates of enteroviruses of pigs. Stages 3-4: To study the relationship selected antigenic strains of enterovirus compared with reference strains antigenic properties enterovirus swine from collection enterovirus swine of immunogenetics laboratory IVM NAAS. Stage 5: Set prametry determination of molecular and genetic properties of enteroviruses of pigs. Methods: processing selected samples of materials by the method of Bogelnd Mayer; selection isolates of these viruses in cell culture CHEB; determining the activity of selected infectious isolates and reference strains of swine enteroviruses in cell culture CHEB titration; the typical association of selected isolates developed by V. Romanenko method (auth. St. Number 1,007,226 on 23 November 1982) in a titration-typed passages in neutralization; determination of neutralization antibody levels in serum. Results:epizootic survey conducted four pig farms enteroviruses prone to diseases of pigs. Selected for serological and virological studies 85 blood samples and 85 samples of rectal swabs of clinically healthy pigs these farms. Of the 85 samples of rectal swabs isolated 20 virus isolates, which were carried out in three successive passages in cell culture CHEB defined their cytopathic and infectious activity in cell culture CHEB. To determine the typical belonging allocated virus isolates was conducted a titration-typed passages with different specific immune sera to enteroviruses pigs of different serotypes. To determine the generic and species belonging allocated isolates viruses studied their physical and chemical properties: sensitivity to air and trypsin, termo esistance (resistance to heating at 56?C for 1 hour without and in the presence of 1 M MgCl2), resistance to environments with the different pH values. Determined immune response to pig farms in these circulating enteroviruses in samples of blood serum neutralization A molecular and genetic evaluation of 20 isolates of enteroviruses of pigs allocated in Ukraine, which had poliantygenetic properties. According to the results of sequencing and phylogenetic analysis of the type referred to PTV I. Developed a new scheme of immunization of rabbits. Reference antigens, worked out and epizootic strains of enterovirus of swine for immunization of rabbits. The obtained serum of rabbits and identified in them antibody titers. Conducted selection of clones reference strains and isolates of enteroviruses of pigs by plaques studied their properties. In addition, the added reference strains of enterovirus swine of National collection of immunogenetics laboratory enteroviruses of pigs, and determined their cytopathic infectious activity in cell culture CHEB Conducted horse reimmunization by antigen production of strain Teschen disease of pigs "Bereznyansky - 652". Obtained highly immune serum for sets diagnostics enzootic encephalomyelitis (Teschen disease) of pigs by methods of neutralization and immunofluorescence Regularly maintained inoculated cell culture SNEV. Laboratory staff participated in the scientific conference of young scientists "Actual problems of veterinary biotechnology and infectious diseases of animals" (m. Kyiv, June 2015). Ukraine received patent number 101,245 for useful model "Method of cultivation swine enteroviruses in cell culture Hep-2", published 08.25.2015 p. In Bull. №16. Approved by the State Veterinary and Phytosanitary Service of Ukraine (protocol № 1 from 12.19.2013 district.) "Methodical recommendations as to cloning swine enteroviruses in cell culture using the method of plaque formation." Developed and approved Methodology Committee (protocol number 2 from 09.21.2015 g.) And IVM Academic Council NAAS (protocol № 10 of 09.22.2015 g.) Methodical recommendations "Getting specific immune serum for the diagnosis of enteroviral diseases swine." Product Description popup.authors Божок В. І. Демиденко І. Ф. Музикіна Л. М. Романенко В. П. popup.nrat_date 2020-04-02 Close