Optimization of Reproduction Stage in Technology of Production of Plague Diagnostic Bacteriophage L-413C

New nutrient media based on baker yeast autolizate were used for the first time for manufacturing of diagnostic preparation of plague bacteriophage L-413C. Experimental media provide high concentration of phage particles at the stage of propagation, and good survivability in lyophilization. Media in which yeast autolizate was a nutrient protein basis appeared to be more effective than those in which it was a stimulating additive. Phage preparations preserved stability of properties during storage at 4-8 °С, and at a higher temperature in the test of accelerated aging. Introduction of yeast nutrient media in technology of plague diagnostic bacteriophage L-413C manufacturing opens good prospects for increasing of production efficiency and decreasing of cost value of the preparatio

LIQUID NUTRIENT MEDIUM FOR SUBMERGED CULTIVATION OF TULAREMIA MICROBE

The paper describes an effective liquid nutrient medium, utilization of which in the process of submerged cultivation of the vaccine tularemia microbe strain allows for the production of high concentrations of viable biomass with low rates of dissociation, which is essential in manufacturing of live vaccines. Dry enzymatic hydrolysate of fibrin, by-product of anti-rabies immunoglobulin production is used as a nutrient-rich base of the new nutrient medium

Enhancement of the Technology for Live Tularemia Vaccine Production

Objective of the study was to develop and test new biotechnological approaches for live tularemia vaccine production.Materials and methods: Francisella tularensis 15 NIIEG strain was used as producer-strain; Francisella tularensis 503 strain – as test infecting one. Producer strain was cultivated on solid and liquid nutrient media. Tangential ultrafiltration was performed with the help of microfiltration module “Viva-flow”. Lyophilization was conducted using drying installation – Free Zone 2.5 L.Results and discussion: Application of the designed liquid nutrient medium on the basis of enzymatic fibrin hydrolysate and submerged cultivation of the producer-strain has allowed for a significant biomass yield increment. At the stage of tularemia microbe culture concentration via microfiltration through filtering membranes with pore size of 0.2 μm, in the mode of tangential liquid flow, increased has been the content of microbe cells; the nutrient media residues – removed. Comparative analysis of the obtained in accordance with experimental technique laboratory series of the vaccine and commercial preparation of live tularemia vaccine has demonstrated their conformity with the specific normative properties. It is established that application of modified liquid nutrient medium, submerged cultivation conditions, methods of biomass concentration and separation has no negative influence on the main properties of live tularemia vaccine and will provide for considerable produce-ability increase in the future

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

Development of Food-Raw-Material-Based Nutrient Media for Submerged Cultivation of Cholera Vibrio Strains

Carried out has been comparative assessment between the performances of liquid nutrient pancreatic fibrin overcook-based and bakery yeast autolyzate-based media and conventionally used in manufacturing of cholera vaccine media for submerged cultivation of Vibrio cholerae 569B and V. cholerae M-41 strains. Results of investigation of media quality biological predictors (morphological and biochemical property stability, efficacy of biomass and protective antigen accumulation), which are fibrin hydrolyzate and bakery yeast autolyzate-based, suggest the possibility of using them for the production of cholera vaccine. Deployment of inedible raw material-based media in manufacturing of cholera vaccine is a prospective technology in view of reduction of medical-prophylactic preparation costs. Moreover it allows for solving the problem of protein waste-product disposal, which is generated in the process of anti-rabies immunoglobulin manufacturing, thus decreasing ecological impact on the environment

NEW NUTRIENT MEDIUM ON THE BASIS OF DRY FIBRIN HYDROLYSATE FOR V. CHOLERAE CULTIVATION

The data on exploration of biological properties of experimental solid and liquid media on the basis of dry enzymatic hydrolysate of fibrinobtained from production waste of anti-rabies immunoglobulin is presened here. The culture media engineered meets the requirements of normative documents and. is highly competitive with the test medium in their qualitative characteristics. Suggested media can be used for V. cholerae cultivation, including submerged cultivation, in production of cholera preventive and. diagnostic preparations

Non-Waste Alternative Technologies in the Production of Heterologous Anti-Rabies Immunoglobulin

Presented is a comprehensive approach to utilization of the wastes that appear in the process of heterologous anti-rabies immunoglobulin production (packed red cells, fibrin, and alcohol-containing products). Specific immunoglobulin is extracted from the surface of red blood cells using desorption technique. Additional yields of immunoglobulin after exposure of erythrocytes to non-ionic detergent amount to 10-19 % of the output. Rich protein supplement feeding for horses-producers is obtained from spray-dried packed red cells. Solid nutritious substrate for microbiological media production is obtained from fibrin using enzymic hydrolysis method. The efficiency of the fibrin hydrolysate-based media is 1.5-2 times higher in comparison with that of the media based on the digest of meat and casein, as demonstrated by the results of Vibrio cholerae scaled cultivation. Furthermore, worked out is the technology of ethanol regeneration after the rivanol-ethanolic precipitation of gamma globulin, alcohol content by volume being (93±1) % after the regeneration. It is demonstrated that the regenerated alcohol can be used as a precipitator in the process of anti-rabies serum fractioning. All in all, the developed techniques make it possible to utilize the wastes of anti-rabies immunoglobulin production and provide for further use of derivatives while producing medical immunobiological preparations

The Efficiency of Baker's Yeast Autolysis Induced by Enzyme Preparations

Has been analyzed the effect of enzyme preparations (trypsin, pepsin, pronase, proteovibrin - enzyme complex isolated from cultural ultrafiltrate of the industrial strain Vibrio cholerae M 41 Ogawa and from non-inactivated autolysate of yeasts) upon the process of autolysis of baker's yeast (Saccharomyses cerevisiae). The selected enzyme preparations being used in small doses render stimulatory effect to autolytic process thus reducing its duration and enabling to obtain autolysate with higher content of amine nitrogen