Abstract
Introduction. When it is nessecery to apply to the antimicrobial elimination therapy, the application of appropriate biological drugs is thealternative treatment. One of them is Biosporin (includes Bacillus subtilis та Bacillus licheniformis). It has a high antagonistic activity against pathogenic and opportunistic pathogenic microorganisms and expresse the immunomodulatory effect. The purpose of our work was to investigate the effect of probiotic Biosporin on different representatives of intestinal microbiota and confirm its activity against opportunistic microorganisms while maintaining the normal symbionts of intestine.
Materials and methods. Experiments were conducted in the albino Wistar rats (males) with initial weighing 180-210 g, which were kept under standard vivarium conditions as required by ethics under the provisions of the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes. Animals were in vivarium under appropriate lighting conditions, temperature and standard diet. During the study they had free access to water. The animals were divided into two groups: 1) control group (n = 12), 2) researched (n = 12). The first group was formed with intact rats. Animals of the second group were given the biological preparation Biosporin in a daily dose 0,2-0,3ml / animal per day. Calculation of doses were performed using doses of individual animals to humans. The biological preparation was administered intragastric every day for 10 days using nontraumatic probe. Faeces were collected on the 10th day into sterile container. When conducting microbiological investigations classic cultural method and differential diagnostic and special nutrient media for the cultivation of certain genera of microorganisms were used. The presence and quantitative values of enterococci, Escherichia, conditionally – pathogenic microorganisms, microaerophilic bacteria – lactobacilli and bifidobacteria were revealed. The results obtained in the investigations of intestinal microflora of experimental rats with the same ones in intact animals were compared. Number of the members of the intestinal microbial landscape was determined by counting of the colonies that grew on media in terms of 1g weight of feces.The conversion to lg of CFU /g (logarithm of the colony forming units in 1 g of faeces) was carry out. The identification of isolated cultures was carried out by morphological, tinctorial, cultural and biochemical properties. State microbiota of the colon was assessed by the population levels of certain species (groups) of microorganisms. Statistical analysis of the results was performed by the method of variational statistic with the determination of the average values of the quantities and the average error. Assessment of the reliability of the differences between average values during the analysis were obtained using unpaired t-Student test. The difference between the values was considered reliable when probability difference p ≤ 0,05.
Results and discussion. Giving the Biosporin to the healthy animals slightly affected the studied quantitative microbial symbionts. Thus, the level of representatives of coccal group (staphylococci and streptococci) was partly higher compared with the intact animals. Staphylococci were found in the number of (5,6 ± 0,48) in experimental animals to (5,2 ± 0,35) lg CFU/ g in control group. Deviation of enterococci compared with control were more significant (9,6 ± 0,60 to 7,0 ± 0,46 lgCFU / g). Streptococci, which do not belong to the genus Enterococcus, were not found in the feaces of intact animals, but after given to rats Biosporin, were found in small amounts (1,2 ± 0,2 lgCFU / g). The highest rate was in Escherichia. These enterobacteria were found in the experiment at 6,35 ± 0,57 lg CFU / g to 5.9 ± 0.40 in control. The absence of Proteus under the influence of Biosporin was noticed, at the time, Proteus mirabilis in the amounts 6,0 ± 0,58 in intact animals was found. Microaerophilic microorganisms (Lactobacillus) in experiment and in intact animals at 7.0 were found out. On the part of bifidoflora some of their elimination, accompanied by a decrease of bifidobacteria compared with controls from 7,5 ± 0,76 to 6,75 ± 0,72 lg CFU / g was recorded. Changes of the amount of yeast fungi Candida in both groups were not recorded.The results of the research of the influence of Biosporin on faecal microbiota of healthy rats is an evidence that the drug causes some redistribution of microbial symbionts. It is clear that in the presence of bacteria – antagonist in the organism of rat the proliferation of investigated microorganisms is unlikely. Increasing of the number of enterococci and streptococci appearance is the result of the movement of microbes from the upper digestive tract, where they are more at its distal. A similar phenomenon also explain an activation of Escherichia in faeces. The number of bifidobacteria does not go beyond the norm. Thus, under conditions of a healthy body Biosporin initiates translocation of symbionts from proximal to distal digestive tract. Moreover, the discharge of natural biological niches promotes renewal of the population. With an excess of opportunistic flora, which includes representatives of the genus Proteus, Biosporin causes their elimination, reflected an appreciable therapeutic effect.
Conclusions. The results of investigations of Biosporin influence on faecal microbiota indicate a redistribution of microbial symbionts. The increase of enterococci in the number and some activation of Escherichiain colon was established. Changes of lactobacilli and bifidobacteria number are insignificant. Obviously, the positive effect of Biosporin is more prominent in the case of dysbiotic changes (excessive proliferation of microflora). The proclivity of quantitative glycolytic microaerophilic microflora decreasing of Biosporin action, is the reason to use the probiotic preparations for reimplantation of lack bifidobacteria and lactobacilli as the next step after Biosporin utilisation.