Comparative evaluation of anti-Helicobacter activity of Stimbifid Plus meta-prebiotic and Helinorm metabiotic

Abstract

Objective. Comparative evaluation of the anti-Helicobacter activity of Stimbifid Plus meta-prebiotic and Helinorm metabiotic and the effectiveness of their oral administration for pathogen eradication in acute experimental helicobacteriosis in conventional white mice, using different microbiological methods. Materials and methods. The following microorganisms were used in the study: Helicobacter H. pylori strain 11, isolated from a biopsy sample of the gastric antral mucosa of a patient with gastritis, and its labeled rifampicin-resistant mutant H. pylori KM-11 (RifR), obtained by spontaneous mutagenesis, capable of growing on a solid culture medium with rifampicin at a concentration of 160 μg·mL–1; probiotic microorganisms – Escherichia coli M-17, Lactobacillus plantarum 8P-A3, Bifidobacterium bifidum No 1. H. pylori and H. pylori KM-11(RifR) were cultivated on a hemin-containing solid culture medium with special additives at a temperature of 37°C using an anaerobic cultivation system (anaerostat). The identification of microorganisms was carried out by morphological features and using bacterial biochemical identification kits. Experimental helicobacteriosis in conventional white mice was formed by oral administration of H. pylori bacterial suspension to animals against the background of the immunosuppressive effect of intramuscular administration of Dexamethasone. Stimbifid Plus meta-prebiotic and Helinorm metabiotic were used in the experiments. Electron microscopy of bacteria was performed using a scanning electron microscope. Statistical processing of the experimental results was carried out according to the Kerber method modified by I.P.Ashmarin and A.A.Vorobyov. Results. In vitro experiments established the anti-Helicobacter activity of Stimbifid Plus meta-prebiotic, which manifested itself as inhibition of the growth of H. pylori bacteria. In parallel experiments, the metabiotic Helinorm did not inhibit the growth of H. pylori bacteria, but only formed co-aggregates with them, diffusing into the culture medium. In experiments in vitro using the disk-diffusion method, evidence was obtained of the actual inhibition of the growth of probiotic microorganisms E. coli M-17, L. plantarum 8P-A3, B. bifidum No 1 by Helinorm metabiotic: L. plantarum 8P-A3 bacteria turned out to be the most sensitive to the effects of Helinorm metabiotic. When studying the survival of H. pylori bacteria in solutions of Stimbifid Plus meta-prebiotic and Helinorm metabiotic at a concentration of 8 mg·mL–1, a pronounced inhibitory effect of Stimbifid Plus meta-prebiotic was established, which manifested itself in the loss of H. pylori viability by 40 minutes of bacterial suspension incubation at a temperature of 37°C, while Helinorm metabiotic only reduced the number of H. pylori bacteria in suspension by 3 orders of magnitude at their initial concentration of 1.2 × 108 CFU mL–1. Experiments on conventional white mice with induced helicobacteriosis against the background of intramuscular administration of Dexamethasone to animals revealed the anti-Helicobacter activity of Stimbifid Plus meta-prebiotic with pathogen eradication by the 9th day of the experiment with absolute preservation of normal gastrointestinal tract biocenosis in experimental animals. In similar experiments on animals infected with Helicobacter pylori, the metabiotic Helinorm did not provide eradication of the pathogen even by the 21st day of the experiment with a significant decrease in the number of leading representatives of the gastrointestinal tract microbiota in infected animals (E. coli, lactobacillus and bifidobacteria). Conclusion. The performed in vitro experiments, as well as helicobacteriosis modeling on conventional white mice, made it possible to study in a comparative way the relationship between the meta-prebiotic Stimbifid Plus and the metabiotic Helinorm with H. pylori pathogen, their influence on the course of the infectious process, prognosis of the disease outcome in infected animals, and the prospects for H. pylori pathogen eradication from animals with oral administration of medications. The results of the experiments confirmed a high anti-Helicobacter activity of Stimbifid Plus meta-prebiotic, as well as the need for further experimental study of the properties of Helinorm metabiotic, proposed by the manufacturer for practical use as the most effective treatment for helicobacteriosis, with consideration of the possibility of correcting the metabiotic structure, or including in its composition additional means that potentiate the anti-Helicobacter activity. Key words: Helicobacter pylori, Lactobacillus reuteri, meta-prebiotic Stimbifid plus, metabiotic Helinorm, helicobacteriosis, eradication