Abstract
The G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology has made a major contribution to the development of probiotic preparations in the Russian Federation. As long ago as the 1960s the researchers at the institute began to conduct investigations on the selection of probiotic strains, development of the probiotic preparations of different formulations and functions and technologies for their industrial production, studies of clinical efficacy and the mechanisms of their action at the cell and molecular levels. At the present time the scientific research and biotechnological activities in this area continue to progress: the scientists at the institute have created a new generation of probiotic preparations, determined new areas for their application, and acquired new knowledge of human microecology.
The G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology pioneered the development of the conception of probiotics in Russia, making it an independent scientific and applied branch, and contributed significantly to the creation of the national arsenal of preparations based on microorganisms, representatives of normal microflora.
The institute began the work in this field in the 1960s. The investigations focused on the selection of probiotic strains from human microbe symbionts, development of probiotic preparations of different formulations and functions, and technologies for their industrial production. The researchers also studied the clinical efficacy and mechanisms of their action at the cell and molecular levels and investigated the effects on the immune and cytokine status and interferogenesis.
According to the traditions of the institute the probiotic investigations were performed in the following areas:
selection of the industrial microorganisms,
maintenance of the bank of microorganisms, representatives of normal flora,
investigations into the nature and mechanisms of probiotic action,
design of means to correct the microflora,
development of industrial technologies,
clinical tests of new preparations and schemes to apply probiotics,
elaboration of criteria for the laboratory assessment of biotope status.
In the above-mentioned period of time the researchers at the institute for the first time developed and advanced techniques to isolate the probiotic cultures from biological material of human origin and mastered their specific identification:
determined the criteria for the selection of promising industrial strains,
formulated the recipes to prepare highly productive and economic nutrient media for both laboratory tests and industrial culture of microorganisms,
developed the formulations and technology to manufacture the first national probiotics and organized their industrial production,
were the first to solve the problem as to how to produce industrial volumes of biomass regardless of the genus, without using vacuum, which allowed unified approaches for designing and producing preparations of different species compositions. Our researchers selected original metabolite cryoprotectors and determined the time and temperature parameters for sublimation and spray drying that allowed production of biomass with high survivability and preservation of microbe cells.
Owing to the results of these studies the researchers singled out one of the priority avenues of the institute's investigations that culminated in experimental and theoretical substantiation for designing pharmaceutical forms of immunobiological preparations of different formulations: tablets, capsules, medicinal suppositories, ointments, creams, and aerosols. At the present time research is being conducted into the creation of injection forms for special spheres of application. The originality of research and developments has been certified with many patents.
In the early 1970s a new branch of microbiological industry was created with the participation and supervision of our institute. By 1990, as a result of these developments, more than 400 million doses of probiotic preparations were produced by a dozen industrial enterprises.
The institute set up the first national collection of bifidobacteria, which served as the basis for organizing the Federal Bifidobacterial Center in 1987. The center was a consultative methodical body and acted as a coordinator of fundamental and applied research that was conducted in this field throughout the country.
In 1994 the Federal Bank of microorganisms, representatives of normal microflora, began to function. The typical strains from the international collections are stored in the bank, which is continuously replenished with new cultures belonging to different systematic groups. This allowed workers to widen the species range of probiotic microorganisms and begin to produce new types of strains and their consortia.
The microbial biopreparations are created using the priority principle of consortium of probiotic strains according to which the synergid interaction of strains is used during the manufacture of the preparation and following persistence in vitro. The competence of this approach is confirmed by the high efficacy of the preparations designed with due regard for the intraspecific synergism. They are the bicomponent preparations Bificol and Bifilong, the three-component Acilact and Kipacid, and the multicomponent preparation Polybacterin. The same principle was used as a basis for formulating functional products Bifilife and the most recent preparations developed by the Institute: biopreparations of synbiotics Bifidum-Multi and Normospectrum based on the consortia of probiotic microorganisms that are complementary to the intestinal microbiocenosis of different human age groups.
The institute has its own clinical facilities and is cooperating with many major national clinics in testing our preparations, which widens the sphere of their application in treating different forms of human pathology. In collaboration with the clinicians in related fields, the researchers at the institute have demonstrated the efficacy of our preparations for treating microecological alterations of different genesis in obstetrics, gynecology, allergology, gastroenterology, hepatology, stomatology, dermatology, and many other fields of clinical practice.
The comprehensive investigations in which different departments of the institute were involved showed that the probiotic preparations have a positive effect on the immune system status, metabolic processes, mineral and vitamin balances, secondary immunodeficiency, and alterations in the production of interferon and cytokines. This greatly extended the sphere of application for probiotics, including prophylaxis and rehabilitation, and gave impetus for developing new forms of probiotics: food additives and functional products.
This new area is the most physiological and effective because it allows design of the recipes enriched with active ingredients that make possible the creation of products with special functions for preventing and treating hypercholesteremia, urolithiasis, bronchial asthma, chronic liver lesions, diabetes mellitus, etc.
Promising lines of research that are being conducted in the G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology are:
combined preparations, food additives and functional products,
multi-species consortia,
consortia with immunoglobulins, interferons, and lysozyme,
consortia with functional ingredients,
metabolite preparations,
autoprobiotics.
The researchers are developing a new generation of probiotics based on metabolites and products produced by the disintegration of cells of productive strains of lactobacilli and bifidobacteria: adhesins, agglutinins, lectins, and high molecular biosurfactants. The investigators continue to develop the lines of research aimed at creating complex preparations based on probiotics and immunoglobulins, interferons, and lysozyme, which are very promising for application in different practical fields of medicine.
A new expanding area of medicinal forms embraces non-traditional bacteria, their components and metabolites, which may be used as probiotics. Thus the preparation Kodivak was developed using cell wall antigens of non-toxigenic corynebacteria. The efficacy of Kodivak was demonstrated by producing normalization of the microflora in the upper respiratory tract mucosa.
During the last 10 years, in collaboration with an academic institute, we have maintained the bank of microbiocenosis for implementing the project ‘Autoprobiotics.’
An important ideological step to extend the application of probiotics was the inclusion of bifidobacteria in the infant's diet and the elaboration of a complex target program for creating adaptive children's milk formulae enriched with bifidobacteria. For this purpose our researchers selected bifidobacteria strains that fermented the milk and using them developed the first children's nutrient formulae enriched with lyophilized bifidobacteria biomass and also the fermented milk bifidumbacterin product.
The clinical observations that were confirmed by the laboratory tests evidenced the beneficial effect of the probiotic cultures on children's organisms that gave impetus to the vigorous development of this branch. At the present time the bifidobacterial products that were developed with the strains and technologies of our Institute are mass produced in milk-processing plants and enterprises manufacturing food for infants.
The institute initiated investigations on human microecology and developed the first methodical aids for bacteriological tests of normal intestinal flora. The results of the investigations in this field are used for publishing and updating at regular intervals the methodical recommendations and standards to determine the qualitative and quantitative composition of microflora of the intestines, upper respiratory tract, and urogenital organs, as well as to diagnose dysbiotic alterations and to develop methods for their correction. These publications are used in their practical work by many laboratories, clinics, and diagnostic centers in the Russian Federation and neighboring countries.
For this purpose in recent years the G.N. Gabrichevsky Institute began to apply biochemical express methods: the enlarged coprologic assay and gas-liquid chromatography to evaluate the metabolic activity of microflora using the composition of short-chain fatty acids in feces. This allows a comprehensive diagnosis of interrelated functional and microecological alterations in the gastrointestinal tract.
At the present time the diagnostic practice is supplemented with a new method called multisubstrate testing, which makes use of an automatic system, Eco-log. This method allows characterization of the microbial community using a multidimensional parameter of functional biological diversity that is the range of substrate consumption that permits monitoring and correction of the dynamics of microbiocenosis.
The comprehensive investigations into characteristics of preparations being developed and new fields for their applications are performed not only by the specialized laboratories, but also by other departments of the institute, which are organically integrated into the research work along with related scientific and medical centers. This allows concentration of the research potential and technical possibilities, and acquisition of new information and significant results.
The important and peculiar feature of the institute is the historically traditional integration of three components – scientific, clinical, and productive facilities that promote the successful development of this branch.
Nowadays the small business enterprises are functioning within the institute, developing technologies for further commercial production of preparations. At the same time the preparations developed by the institute are also manufactured by many Russian enterprises.
The consultative diagnostic center that was set up in the institute has acquired a reputation as one of the most competent centers in Moscow, owing to its effective medical treatment. This has its origins in the experience and traditions accumulated in the institute and in the quest for higher professional qualifications, which was always an essential requirement self-imposed by every worker at the institute.