Introduction . The role of the respiratory tract microflora violations as an etiological and pathogenetic factor in the development of the inflammative upper respiratory tract disease is indisputable, and the importance of persistence factors of respiratory opportunistic microorganisms (URT) is high. The traditional use of antibiotics and antimycotic agents in the pathology of URT has failed due to a significant increase in the number of antibiotic-resistant strains of microorganisms and the development of a number of undesirable adverse reactions in patients. An alternative advantage in these conditions is belonged to herbal preparations. Aim . To conduct a comparative in vitro study of antibacterial and antifungal activities of the plant essential oils as a component of the Dyshi Oil composition and the Dyshi Oil composition itself against opportunistic microorganisms - causative agents of inflammatory upper airway diseases. Materials and methods . Essential oils of clove, juniper, peppermint, eucalyptus, as well as the composition of oils “Dyshi” were used in the work. Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ICIS-278, Pseudomonas aeruginosa ATCC 27853, and Candida albicans ATCC 24433 bacterial cultures were used as test cultures. Staphylococcus epidermidis 25 strain, isolated from a conditionally healthy person, was chosen as a representative of the URT normobiota. Antimicrobial activity of essential oils was determined by the method of diffusion in nutrient agar, MIC - by the method of serial dilutions. Results . The presence of zones of pronounced growth retardation of Staphylococcus aureus , Klebsiella and Pseudomonas , as well as yeast-like fungi was established when exposed to the composition of “Dyshi” oils, while separately used oils did not have a visible effect on the growth of bacteria and fungi, which indicates a synergistic interaction of the drug oils components. Conclusion . The presence of a synergistic antimicrobial effect of the “Dyshi” oil composition against opportunistic bacteria and yeast-like fungi of the genus Candida has been proven. The absence of an inhibitory effect on the growth of Staphylococcus epidermidis, which is a representative of the upper respiratory tract normobiota indicates a possible selective effect of “Dyshi” oil aimed at maintaining the colonization resistance of the URT biotope.
Introduction. The work includes the results of clinical and microbiological studies that determine the feasibility of using the herbal preparation EPS® 7630 for the treatment of rhinopharyngitis (RF) in children. The introduction provides a brief review of the literature data on the role of the microbial factor and the important contribution of dysbiotic disorders of the URT microflora in the development of the Russian Federation. The porpose of persistent properties of opportunistic microorganisms (UPM) is outlined: the ability to biofilm formation – BPO and inactivation of lysozyme – ALA as dysbiotic markers of RF, which create conditions for relapses and chronicity of the disease, reduce the effectiveness of antimicrobial therapy. Goal.The aim of the work was the clinical and microbiological substantiation of the effectiveness of using EPS® 7630 in the treatment of RF in children.Materials and methods. The study involved 79 children aged 3 to 15 years. 1st group – 43 children who received treatment, including, along with traditional treatment, the drug Umckalor EPS® 7630, 2nd group (comparison) – 36 children who did not receive the tested drug. Microbiological examination was carried out by taking swabs from the pharynx and nose before and after treatment; BPO and ALA were determined according to the methods of Bukharin O.V. et al. (1999).Results. Conducted clinical studies have established a reduction in the timing of relief of the main symptoms of RF (cough, rhinitis, sore throat, fever) in children treated with EPS® 7630 compared to the comparison group. Microbiological studies of microorganisms isolated from the nasopharynx of children with RF revealed the ability of EPS® 7630 to suppress bacterial persistence factors (BPO and ALA) by 60% or more from the initial level.Conclusions. The data obtained reveal one of the mechanisms of the antimicrobial action of EPS® 7630, which consists in suppressing the persistent potential of UPM and regulating the effect of the drug on the microbiocenosis of the URT. The clinical and microbiological efficacy of using EPS® 7630 in the treatment of RF in children has been proven.
Based on the clinical and microbiological monitoring of two groups of children aged 3 to 17 years with acute (n=78) and chronic (n=46) course of reactive arthritis (ReA), a method for early diagnosis of chronic arthritis was developed by determining the number of antibiotic-resistant coprostrains in patients with ReA, characterized by the absence of the need to isolate a pure culture of the pathogen and its identification; inoculation of faeces at a dilution of 10-5 on solid 1.5% GRM-agar with an antibacterial agents used in the treatment of a particular patient, at a minimum inhibitory concentration in the resistance range, followed by incubation and counting of the colonies of microorganisms grown on the plate. A significant relationship between the number of antibiotic-resistant gut bacterial strains and the course of arthritis (acute, chronic) was revealed, and the borderline value of the number of antibiotic-resistant gut bacterial strains was determined - 5×103 CFU/g, which allows differentiating the acute course from the chronic one: in the acute course< 5×103 CFU/g, with chronic - ≥ 5×103 CFU/g. The method allows, at the stage of completion of anti-inflammatory therapy in the active phase of the disease, to identify a risk group for the development of a chronic course of arthritis among patients with ReA, which can contribute to timely therapeutic measures aimed at preventing recurrence of the disease and making the patient disabled.
Analysis of the study is to assess the diagnostic significance of cytokines in the sperm plasma of men of reproductive age (20 - 45 years) of two groups: of patients with chronic bacterial prostatitis, not complicated by infertility and with loss of fertility. The study of sperm plasma - the WHO standard. Determination of the level of cytokines in seminal plasma - by enzyme immunoassay («Cytokine», Russia). Two methods of mathematical statistics were used: discriminant analysis and classification trees (decision trees).The similarity of interpretations of discriminant analysis and decision tree was noted, where the main role in both cases belongs to the cytokine IL-4. The level of sperm IL-4 in combination with therapeutic monitoring can be used for the medical management of patients with chronic prostatitis in order to prevent the development of infertility and to develop methods for screening diagnostics of fertility disorders in men.
Introduction. The results of numerous studies indicating the relationship between the development of reactive arthritis (ReA) and the disturbance of the intestinal microbiocenosis give rise to interest in commensal microorganisms that make up the intestinal microbiota as potential initiators of immune-dependent inflammatory diseases of the joints. In this regard, the question of the expediency of using probiotic preparations for the correction of ReA patients intestinal microflora is of practical interest.The purpose of the study was to evaluate the effectiveness of Lactobacillus rhamnosus LGG (LGG) in the preventive treatment of children with ReA to prevent the development of relapses.Materials and methods. Clinical and microbiological study included 60 patients with ReA from 3 to 17 years old, who were divided into two groups, 30 children each. Patients of the group 1 received courses of treatment with LGG in the inactive phase of the disease. Patients of the group 2 (comparison group) were not treated with probiotic. The criteria for the effectiveness of treatment were the number of relapses of ReA during follow-up observation for 1 year; dynamics of intestinal microbiocenosis condition according to the following parameters: indicator of microbial contamination (IMC) of intestinal microsymbionts; ability to biofilm formation (BF); levels of lactoferrin and lysozyme in coprofiltrates.Results. In patients treated with LGG, relapses of arthritis were significantly less frequent during 12 months of prospective observation relative to the comparison group. Patients of the group 1 showed positive dynamics of the state of intestinal microbiocenosis: a decrease in the severity of dysbiosis, a decrease of lactoferrin and lysozyme level in coprofiltrates, IMC and BF of opportunistic microorganisms against the increase IMC and BF in bifidobacteria and lactobacilli.Conclusions. The use of LGG in the treatment of children with ReA in the inactive phase of the disease contributes to the correction of intestinal microbiocenosis disorders and reduces the number of arthritis relapses.
The authors present the results of the study of one of the basic physiological properties of microorganisms of intestinal microbiota — the ability to biofilm formation (BF) in children with reactive arthritis. It was shown that BF is a biological marker which determines the arthritogenicity of microorganisms of the intestinal microbiota and can serve as a biological target for the choice of effective therapy in arthritis.
The article summarizes the results of the work of domestic and foreign researchers on the study of homeostasis of the nasal cavity, including the state of the microbiota of the nasal mucosa in healthy children and in inflammatory diseases of the upper respiratory tract. The normal microbiota of the nasal cavity is represented by corynebacteria (diphtheroids), neisseria, coagulasenegative staphylococci, alpha-hemolytic streptococci. From among the transient species, such species as Staphylococcus aureus, Escherihia coli, beta-hemolytic streptococci would be found in the specified biotope. The main attention is paid to the features of the nasal microbiota in newborns and young children (dependence of the nasal microbiota on the type of feeding, age and season). The role of hypercolonization by opportunistic microflora and its persistent potential (ability to biofilm formation, degradation of lysozyme, interferon) in the pathogenesis of inflammatory diseases of the upper respiratory tract is noted. The pathogenetic mechanisms providing multilevel protection of the organism from pathogens are considered. The expediency of using elimination-irrigation therapy as a hygienic means of sanitizing the nasal cavity in young children has been determined. The questions about the advantages of the use of isotonic solutions, their effectiveness for ensuring normal colonization resistance of the nasal mucosa are discussed. In conclusion, we must say that for hygienic care and sanitation purposes it is recommended only to rinse the nasal mucosa in newborns and infants using drops of isotonic saline solution and an aspirator to avoid aspiration and inflow of liquid into the cavity of Eustachian tube and middle ear, and also for the preventive purposes, as it allows to maintain homeostasis of the nasal mucosa and prevent the development of acute respiratory diseases.
The multidirectional effect of recombinant cytokines (TNF-α, IL-10) on the biofilm formation of intestinal microsymbionts was established, which depended on the concentration of cytokines, the type and strain of microorganisms.
A simple accelerated method for determining «self/non-self» microorganisms by using the agglutination reaction (RA) and therapeutic/prophylactic serum (Immunoglobulin complex preparation, lyophilized IgG, IgA, IgM immunoglobulins, developed by CSC Immuno-Gem, Moscow) is proposed to test for pathogenic, opportunistic and dominant probiotic Bifidobacteria spp. In parallel, all the microbial cell cultures examined were registered in the databases of Russia-wide and international collections and tested by the intermicrobial “self/non-self” recognition method, previously developed by us. 16 collection strains of various microorganisms were assessed by the RA with relevant therapeutic and prophylactic serum. Biological samples were obtained from the collection bacterial strains of Bifidobacterium bifidum 791, Escherichia coli LEGM-18, Klebsiella pneumoniae 278, Lactobacillus fermentum 90T-C4, Bifidobacterium longum MC-42, Escherichia coli M-17, Shigella sonnei 177b, Shigella flexneri 170, Escherichia coli 157, Staphylococcus aureus 209, Candida albicans 10231 and Salmonella serovar Enteritidis ATCC 10708. In addition, cell cultures obtained from the Museum of the Institute of Cellular and Intracellular Symbiosis UB RAS such as Bifidobacterium longum ICIS-505, Lactobacillus acidophilus ICIS-1127, Bifidobacterium bifidum ICIS-202, Bifidobacterium bifidum ICIS-310 were also included into the study. To assess microbial peptidoglycan foreignness, the intermicrobial “self/non-self” recognition method was also used based on inducing metabolites produced by the “dominant” test strain Bifidobacterium longum MC-42 after pre-incubation with metabolites collected from the studied cell cultures (“associates”) followed by established “dominant-associate” feedback loop. The data were evaluated by assessing change-fold in reproduction (growth/replication) and adaptation (biofilm formation and anti-lysozyme test) of microbial cultures in accordance with the described technique followed by comparing these two methods for intermicrobial “self/non-self” recognition. All the RA data were found to fully agree with those obtained after previous studies by using intermicrobial “self/non-self” recognition method coupled to “dominant-associate” system. Moreover, compared to analogous “intermicrobial recognition” method (5 days), ease of use and test timeframe (24 hours) allow to consider RA attractive for screening studies to select strains for scientific and industrial purposes.
This report presents the data on the draft genome sequence of Bifidobacterium bifidum strain ICIS-202. The strain, isolated from the intestine of a young healthy woman, was deposited in the State Collection of Microorganisms of Normal Microbiota in Gabrichevsky Institute of Epidemiology and Microbiology, Moscow, Russian Federation as a prospective candidate for probiotic development. The size of the genome was 2,265,060 bp (62,4% G + C content). The annotation revealed 1771 coding sequences, including 1771 proteins, 5 rRNA, 52 tRNA, and 3 ncRNA genes. The draft genome sequence data of B. bifidum strain ICIS-202 is available in DBJ/EMBL/GenBank under the accession nos. SSMS00000000.1, PRJNA412271 and SAMN07709009 for Genome, Bioproject and Biosample databases, respectively.
This report describes the genome sequence of Bacillus paranthracis strain ICIS-279, isolated from human feces. It demonstrates a tumor necrosis factor alpha (TNF-α) inhibitory activity up to 0.1 ng/ml. The genome size is 5,180,499 bp, with a G+C content of 35.4%. Annotation revealed 5,168 coding sequences, including 5,168 proteins and 43 rRNA, 102 tRNA, and 5 noncoding RNA (ncRNA) genes.
Aim. To study in comparison immunoregulatory properties of dominant and associative microsymbionts metabolites in human large intestine’s eubiosis and dysbiosis. Materials and methods. 260 strains of bifidobacteria used as dominant microbiota, 132 cultures of conditionally pathogenic bacteria and fungi used as associative microsymbionts from 122 intestinal microsymbiocenoses. The cytokines production was studied in cultures of mononuclear cells co-cultivated with microsymbionts’ supernatants. The results were processed statistically (Statistica 10.0). Results. In eubiosis, dominant and associative microsymbionts showed immuno regulatory properties heterogeneity. In the case of phlogogenic cytokines, the associates equally exhibited stimulation / suppression / no effect on cytokines, except for enterococci and bacteroids, stimulating IL-8 secretion, and lactobacilli, inducing IFNy. Dominants were characterized by a unidirectional effect: IL-10 secretion stimulation and TNFa, IFNy and IL-17suppression, while retaining the induction of IL-10 in dysbiosis. In contrast, supernatants of the associates combined the opposing cytokines production: the early proinflammatory cytokine TNFa, the immunoregulatory cytokine IFNy and the antiinflammatory cytokine IL-10. Conclusion. Intestinal homeostasis in eubiosis is supported by differentiated effects of microsymbionts’ metabolites on the production of antiinflammatory, immunoregulatory cytokines with the formation of an optimal balance, limiting inflammatory and autoimmune reactions. The dominance of the immunoregulatory properties remains intact in the conditions of dysbiosis, and the variety of effects on pro-/antiin-flammatory cytokines is limited in the associates.
Aim. To study in comparison immunoregulatory properties of dominant and associative microsymbionts metabolites in human large intestine’s eubiosis and dysbiosis. Materials and methods. 260 strains of bifidobacteria used as dominant microbiota, 132 cultures of conditionally pathogenic bacteria and fungi used as associative microsymbionts from 122 intestinal microsymbiocenoses. The cytokines production was studied in cultures of mononuclear cells co-cultivated with microsymbionts’ supernatants. The results were processed statistically (Statistica 10.0). Results. In eubiosis, dominant and associative microsymbionts showed immuno regulatory properties heterogeneity. In the case of phlogogenic cytokines, the associates equally exhibited stimulation / suppression / no effect on cytokines, except for enterococci and bacteroids, stimulating IL-8 secretion, and lactobacilli, inducing IFNy. Dominants were characterized by a unidirectional effect: IL-10 secretion stimulation and TNFa, IFNy and IL-17suppression, while retaining the induction of IL-10 in dysbiosis. In contrast, supernatants of the associates combined the opposing cytokines production: the early proinflammatory cytokine TNFa, the immunoregulatory cytokine IFNy and the antiinflammatory cytokine IL-10. Conclusion. Intestinal homeostasis in eubiosis is supported by differentiated effects of microsymbionts’ metabolites on the production of antiinflammatory, immunoregulatory cytokines with the formation of an optimal balance, limiting inflammatory and autoimmune reactions. The dominance of the immunoregulatory properties remains intact in the conditions of dysbiosis, and the variety of effects on pro-/antiin-flammatory cytokines is limited in the associates.
The comparative analysis of content of cytokine and significant local antimicrobial factors (IL-6, IL-8, IL-17, INF-γ, TNF-α, IL-10, CRP, lysozyme, lactoferrin) in the coprofiltrates and serum of patients with acute and chronic reactive arthritis (ReA) made it possible to determine the markers predicting the course and outcomes of arthritis. It is established that the method of predicting ReA involving the analysis of the parameters characterizing coprofiltrates of children is the more accurate and effective method.
The authors present the results of the study of one of the basic physiological properties of microorganisms of intestinal microbiota — the ability to biofilm formation (BF) in children with reactive arthritis. It was shown that BF is a biological marker which determines the arthritogenicity of microorganisms of the intestinal microbiota and can serve as a biological target for the choice of effective therapy in arthritis.
The comparative analysis of content of cytokine and significant local antimicrobial factors (IL-6, IL-8, IL-17, INF-γ, TNF-α, IL-10, CRP, lysozyme, lactoferrin) in the coprofiltrates and serum of patients with acute and chronic reactive arthritis (ReA) made it possible to determine the markers predicting the course and outcomes of arthritis. It is established that the method of predicting ReA involving the analysis of the parameters characterizing coprofiltrates of children is the more accurate and effective method.
Aim. To study the spectrum and level of short-chained fatty acids (SCFA) in supernatant of bifidobacteria under different microecological conditions of the colon biotope in human. Materials and methods. Metabolites of 88 bifidobacteria strains isolated from patients when examined for dysbiosis of the colon were investigated. Definition of concentration of SCFA was performed on acidified supernatant samples by a separation method on chromatograph GC-2010 Plus, Shimadzu (Japan). Results. Monobasic acids were found in metabolites of 50 -100% study cultures of bifidobacteria where the spectrum and level of carboxylic acids in supernatants varied depending on microecological condition of the origin of the discharge. In severe damages of microsym-biocenosis in metabolites of Bifidobacterium spp., summarized concentrations of SCFA, structural index, levels of aceitic and propionic acids were decreased. Strain-specific differences in a metabolic profile of bifidoflora in a composition of individual consortiums were determined. Data obtained indicate the variation of functional (metabolic) activity of dominant strains in different microecological conditions of the human colon. Conclusion. Uniquieness of metabolome of every other strain due to their strain specifity determines their functional activity, but a metabolic profile of bifidoflora can serve as the most important criterion for the selection of effective probiotic drugs for treatment and prevention of dysbioisis in the colon.
Acute generalized infection caused by P aerugenosa in mice (CBAxC 57 Bl 6 )Fj, is characterized by more severe morphological and functional disorders than the same infection caused by K. pneumoniae. The use of miliacin has a protective effect in case of sepsis, makes structural disorders less severe and modulates functional activity of immune peritoneal macrophages.