THE DEVELOPMENT OF MOLECULAR SPECIES TYPING SYSTEM FOR IXODES PERSULCATUS AND IXODES PAVLOVSKYI TICKS

Determination of ticks species is very important for epidemiology and ecology investigations but in some cases the morphological study couldn't be carried out. In this work the PCR system based on species-specific primers to cytochrome oxidase subunit 1 (COI) gene sequence was developed for molecular species typing of Ixodes persulcatus and. Ixodes pavlovskyi ticks

Tickborne Pathogen Detection, Western Siberia, Russia

Ixodes and Dermacentor ticks harbor Borrelia, Anaplasma/Ehrlichia, Bartonella, and Babesia species

EFFECT OF THE PROPHAGE CTXΦ DELETION UPON PHENOTYPIC PROPERTIES IN STRAINS OF VIBRIO CHOLERAE BIOVAR EL TOR, ASSOCIATED WITH VIRULENCE AND PERSISTENCE

Objective of the study is to evaluate the influence of CTXφ prophage deletion, which carries ctxAB genes, on phenotypical properties associated with pathogenicity or biofilm formation in non-toxigenic mutants. Materials and methods. Utilized have been the clinical strains of Vibrio cholerae biovar El Tor and their spontaneous non-toxigenic mutants that lost CTXφ prophage. Applied have been microbiological and biochemical methods, inoculation of model animals with cells of the strains under study. Results and conclusions. The results of comparative analysis of phenotypic properties in isogenic toxigenic and non-toxigenic strains of Vibrio cholerae biovar El Tor, which lost CTXφ prophage encoding the cholera toxin, are represented. It is established that the deletion of CTXφ prophage leads to the simultaneous change of several phenotypic properties associated with virulence (colonizing ability, production of soluble hemagglutinin/protease and heat labile hemolysin/cytolysin) and biofilm formation (motility, exopolysaccharide biosynthesis) in spontaneous non-toxigenic mutants. It is suggested that the reason for these phenotypic changes in the mutants might be the changes in activity of the related to each other regulatory genes controlling virulence and biofilm formation process in cholera agent

Anesthesia of laboratory animals in manufacturing of diagnostic and preventive biomedicines

Preparations such as XilaVet, Zoletil 100 as well as Aeranne (Isoflurane) are successfully applied for animal anesthesia in veterinary practice. We assessed a possibility of using parenteral narcosis with Zoletil 100 in combination with muscle relaxant Xila for producer-rabbits involved in manufacturing of natural rabbit serum subsequently deployed for production of diagnostic serum and immunoglobulin preparations. Administration of preparations into auricular vein is easy to do, while animals are sedated immediately allowing for safe fixation on restraining table and causing no additional stress for biomodels. This type of narcosis provides for expected depth of anesthesia and its maintenance until the end of blood-letting procedure. Parameters characterizing the state of cardiovascular system due to anesthetic products remained within the permitted limits. These preparations do not reduce heart beat rate allowing for collecting sufficient blood volumes. Application of inhalation anesthesia with Aeranne in laboratory animals provides for the specified depth of anesthesia and its maintenance until the end of the whole procedure. However, it requires specialized equipment and highly trained personnel with appropriate skills. Usage of Xila as a mono narcosis is not recommended as exhibits weak analgesic effects and strong hypotensive activity by decreasing quantity of collected blood volume. It was found that anesthetics such as Xila, Zoletil 100, Aeranne did not affect specific activity of immune sera in case of total dehematizing procedure. Moreover, antibody titers were not declined throughout entire observation (12 months) period and complied with the requirements of regulatory documentation. In addition, a feasibility of replacing old-fashioned anesthesia method with diethyl ether for a combination of safer contemporary preparations of Zoletil 100 and Xila was demonstrated while manufacturing tableted chemical cholera vaccine in experimental series with suckling rabbits used at diverse stages of raw material verification during surgical interventions. Xila, Zoletil 100, and Aeranne examined by us had no impact on the amount of blood obtained from donor-animals, immunological properties of the sera and ready-to-use diagnostic preparations. Such drugs were safe for all-age animals that comply with the requirements to anesthesia of animal biomodels and producer-animals in manufacturing of immunobiological preparations. Thus, our study allowed to conduct experiments with laboratory animals in a more humane manner

Разнообразие Borrelia burgdorferi sensu lato в природных очагах Новосибирской области

PCR assays were used to test sample from Ixodes persulcatus, blood and tissues of small mammals, human blood after tick bi- tes, as well as isolates from adult ticks. It was demonstrated the presence of two Borrelia species: B. garinii and B. afzelii. Mainly DNA B. garinii NT29 were determined.Методом ПЦР исследованы образцы от Ixodes persulcatus, кровь и ткани органов мелких млекопитающих, образцы крови человека, а также изоляты боррелий от голодных имаго. Показано, что на территории Новосибирской области рас- пространены боррелии двух видов — B. garinii и B. afzelii. Образцы, содержащие ДНК B. garinii подгруппы NT29, преобла- дали среди исследованных

Usage of nutrient Medium Based on Dry Hydrolysate of Casein in Manufacturing Bivalent Chemical Cholera Vaccine

Objective of the study was to select the standardized substrate containing dry hydrolysate of casein for preparation of nutrient medium utilized for manufacturing bivalent chemical cholera vaccine under submerged cultivation of cholera vibrio strains in fermenters. Materials and methods. We used Vibrio cholerae O1 strains of classical biovar: strain 569B Inaba and strain M-41 Ogawa. Examined were two dry substrates of the medium: enzymatic hydrolysate of casein, Type I Himedia (India) and pancreatic hydrolysate of casein, produced by the State Scientific Center of Applied Microbiology and Biotechnology (Russian Federation). Produced under laboratory conditions at the premises of the RusRAPI “Microbe” medium was used as a control. Submerged cultivation was conducted in bioreactors during (9±1) h with aeration and automatic feeding of glucose and ammonia. Production of protective antigens was measured applying immunochemical and biological methods. Results and discussion. It is demonstrated that submerged cultivation of cholera vibrio production strains on nutrient media under study provides for synthesis of protective antigens the parameters of which comply with the requirements of normative documentation. More standardized and higher indicator values of the target product are ensured by cultivation of producer strains on nutrient medium with a substrate from dry enzymatic hydrolysate of casein, containing (1.5±0.1) g/l of amino nitrogen for the strain V. cholerae M-41 and (2±0.1) g/l – for V. cholerae 569 B. Transition to the use of standardized dry protein components of cultivation media does not lower the quality of the chemical cholera vaccine, but allows for the reduction of cost price and duration of technological process

Improvement of Technology of Cholera Toxin B-Subunit Production

Consideration is given to implementation of state-of-the-art filtration technologies for up-scaled manufacturing of cholera toxin B-subunit, produced by recombinant Vibrio cholerae non O1 KM93 strain. Selected are micro- and ultra-filtration membranes to be incorporated into manufacturing method. Investigated are the properties of cholera toxin B-subunit, obtained applying the pilot technology. The engineered method for up-scaled manufacturing of cholera toxin B-subunit makes the procedure easier-to-maintain due to tangential micro- and ultra-filtration, performed at the stage of purification and concentration. It excludes labor-consuming chromatographic purification, while retaining B-subunit properties. The studies undertaken make it possible to manufacture cholera toxin B-subunit with the same characteristics as in the case of the pilot technology, but under production conditions, and use it as a component for chemical cholera vaccine

Об эффекте взаимодействия хлорсодержащих и азотсодержащих полимеров в адгезионных композициях

The opportunity of modification of adhesives based on nitrile-butadiene rubber with chlorinated PVC is shownРассмотрены особенности адгезионных свойств клеев на основе азотсодержащих эластомеров и хлорпроизводных полимеров

Properties of Experimental and Standard Preparations of the Protective O Antigen of <I>Vibrio cholerae</I> M41 Ogawa

Investigated are immunochemical, chemical and biochemical properties of the O-antigen preparations obtained from Vibrio cholerae M41 classical biovar, serotype Ogawa strain using standard manufacturing and improved technologies of their concentrating. The preparations have been concentrated by ultrafiltration in the mode of tangential liquid flow with membranes of various cut-off levels for a particular molar weight. ELISA has revealed equal O-antigen load in all of these preparations. Studies of specific fractures by means of chromatographic and electrophoretic methods have demonstrated their properties to be alike. Chemical composition analysis has also identified coincidence in the load of components under specification. Thus, it has been proved that application of the improved V. cholerae protective antigen concentrating technology for manufacturing cholera chemical bivalent palletized vaccine is justified