Aim. To determine the composition and properties of the gut microbiota in HIV-infected patients with pulmonary tuberculosis.Materials and Methods. We studied 92 faecal samples from patients with pulmonary tuberculosis (n = 46) and patients with combined pulmonary tuberculosis and HIV infection (n = 46), with the following examination of the appearance, cultural properties, and biochemical profile of the bacteria. The constancy of microbial taxa was determined using Dazho-Odum indicator. Microorganisms were classified as constant at C > 50%, as complementary at 25% < C < 50% and as occasional at C < 25%.Results. Gut microbiota of patients with multidrug-resistant tuberculosis was consistently represented by Bifidobacterium spp., Lactobacillus spp., Enterococcus spp., Escherichia spp., Staphylococcus spp., and Candida regardless of theirHIV status. Species composition and prevalence of virulence factors in Staphylococcus spp. and fungi isolated from patients with multidrug-resistant tuberculosis also did not depend on HIV status. Complementary microorganisms were represented exclusively by Clostridium spp., while random microbiota was represented by 6 genera (Enterobacter spp., Citrobacter spp., Salmonella spp., Klebsiella spp., Proteus spp., and Pseudomonas spp.) belonging to the Proteobacteria phylum.Conclusion. Similar composition of gut microbiota in HIV-positive and HIV-negative patients with multidrug-resistant tuberculosis indicates common mechanisms of intestinal dysbiosis and a uniform approach for its correction.
The aim: In vitro identification of targets for antagonism factors in klebsiellas and enterococci for Candida albicans isolated from the intestinal microbiome of HIV infected patients.Materials and methods. The tests were performed using 38 Candida albicans strains, 28 Klebsiella pneumoniae strains, and 30 Enterococcus faecalis strains isolated from the intestinal microbiome of 89 HIV infected children. The mean age of the patients was 24 ± 2 months; the group consisted of 49 (55%) boys and 40 (45%) girls. Microorganisms were isolated from the intestinal biotope using such selective media as HiChrome Candida Agar, HiChrome Klebsiella Selective Agar Base, and Enterococcus Agar; the study included identification of species. Model experiments were performed to study anti-catalase activity of E. faecalis exometabolites and the impact of K. pneumoniae on morphological transformation of C. albicans fungi.Results. Klebsiellas decrease the intensity of germ tube formation in C. albicans by 58.7% (p < 0.01). When cocultured, 12.3% of the yeast cells produce germ tubes, while 29.8% of transformed cells was detected in the fungal monoculture. It has been found that exometabolites of 65.7% of E. faecalis strains decrease production of catalase in C. albicans. The initial catalase level in untreated cultures of C. albicans averages 1.02 µmol/min of optical density; after they are treated with E. faecalis exometabolites, the level decreases to 0.55 µmol/min, i.e. by 46.1% (p < 0.05).Conclusions. K. pneumoniae and E. faecalis demonstrate antagonism of different intensity toward C. albicans. Morphological transformation and catalase production are targets for antagonism factors of facultative microbiota in C. albicans.
Fungi are opportunistic microorganisms that colonize all biotopes of the human body, including intestinal. In case of emerging adverse environmental factors (HIV infection, other immunodeficiencies, antibiotic therapy), these microbial representatives begin active reproduction, which might require prescribing antimycotics. Frequent use of the latter in clinical practice induces the development of drug resistance to antifungal drugs, which may impact on effectiveness of both the treatment of fungal infections and other diseases. The purpose of the study was to assess the pattern and spectrum of drug resistance of Candida genus in the intestinal biotope of patients with respiratory tuberculosis and identify risk factors for developing total fungal drug resistance to antimycotic drugs. Material and methods. There were enrolled 21 patients with respiratory tuberculosis. Pattern of the fungal species diversity for the Candida genus isolated from faeces was evaluated, and the spectrum of relevant drug resistance to antimycotic drugs was determined. Patients (n=21) were divided into 2 groups: with (n=10) and without (n=11) total resistance to antimycotics, after which the main risk factors for its development were identified. Results. Members of the Candida genus were isolated from all patients examined, wherein pure cell cultures were characterized by high level of antimycotics resistance. Resistance to three drugs was noted in 1 culture (4.8%), to four in 10 cultures (47.6%), also found in 10 cultures to the entire drug panel (47.6%). During statistical processing, the data were obtained on affecting formation of total resistance to antimycotics of concomitant pathology of the gastrointestinal tract, the presence of a clinically significant dyspeptic syndrome, a history of antimycotic therapy, HIV infection with severe immunodeficiency, and some decrease in the peripheral blood CD4+ lymphocyte count. Conclusions. Fungi of the Candida genus isolated from tuberculosis patients were characterized by high level of resistance to antimycotics. Total resistance was observed in 47.6% of patients. In addition, the major fungi colonizing the intestines of tuberculosis patients were found to be Candida albicans species. The risk factors for the development of total antifungal resistance included: chronic enterocolitis, dyspeptic syndrome, peripheral blood CD4+ lymphocyte count lower than 350 cells/l, and history of antimycotic therapy.
The article describes risk factors for the development of intestinal indigestion syndrome (IIS) in 109 patients with tuberculosis while they were receiving anti-tuberculosis chemotherapy (ATCT) and the results of assessment of the intestinal microbial population in 30 patients with tuberculosis before chemotherapy start. The following factors made significant contribution to development of intestinal indigestion syndrome: HIV infection, the number of doses of anti-tuberculosis drugs taken, and chronic gastrointestinal diseases. Even before the start of chemotherapy, the patients had disorders of the intestinal microflora, which were characterized by a decrease in the quantity of obligate bacteria, and higher frequency and number of facultative and transient bacteria.
Aim. As the activity of bacteriophages is species- and strain-specific, it is necessary to study bacteriophage sensitivity in distinct geographic regions with various disease patterns. Here, we aimed to study the lytic activity of specific commercially available bacteriophages against Klebsiella spp., Proteus spp., and Staphylococcus aureus isolated from the intestines of children with gut dysbiosis.Materials and Methods. Bacteriophage sensitivity was assessed in 315 opportunistic microorganisms (125 Staphylococcus aureus strains, 120 Klebsiella spp. strains, 70 Proteus spp. strains) isolated from the intestinal microbiota of 300 children < 4 years of age with gut dysbiosis. Bacteriophage preparations were produced by Microgen (Russian Federation). The lytic activity of bacteriophages was studied by a drip method on a Muller-Hinton medium by calculating the area of bacterial culture lysis.Results. We found low sensitivity of Klebsiella spp. (37.5% sensitive strains) and Proteus spp. (41.4% sensitive strains) to specific bacteriophages, albeit there were considerable differences between distinct Klebsiella species (Klebsiella pneumoniae, 56.7% sensitive strains; Klebsiella oxytoca, 18.3% sensitive strains, p = 0.03) and Proteus species (Proteus vulgaris, 52.0% strains; Proteus mirabilis, 35.6% strains, p = 0.04). Nevertheless, sensitivity to Staphylococcus aureus was considerably higher (78.4%). In addition, lytic activity of bacteriophages reduced along with the increasing severity of gut dysbiosis.Conclusion. Klebsiella spp. and Proteus spp. isolated from children with dysbiosis have low sensitivity to commercially available bacteriophages. Bacteriophage sensitivity positively correlated with gut dysbiosis.
The progress in investigating microbiome in children and adolescents as well as its impact on maintaining health is considered one of the most significant achievements of modern medicine. In children with various diseases, the qualitative and quantitative indicators of the intestinal microflora change, which causes the development of dysbiosis. The assessing changes in intestinal microflora in children during treatment with anti- tuberculosis drugs has retained its importance and relevance due to insufficient data in the field, which requires a more detailed understanding of this problem. The presented article assesses the state of gut microbial biocenosis in children receiving anti-tuberculosis treatment. The purpose of the study: to study the microbial landscape of the colonic contents in children receiving anti-tuberculosis therapy. Materials and methods. The study included 24 children with verified tuberculosis of the respiratory organs receiving treatment in a 24-hour hospital of the Kuzbass Clinical Phthisiopulmonological Medical Center named after I.F. Kopylova. The study of intestinal microflora was carried out by a quantitative bacteriological method. The material of the study was feces collected in a sterile disposable container, which was delivered to the Department of Microbiology, Immunology and Virology of the Federal State Budgetary Educational Institution of the Russian Ministry of Health. Results. Dysbiosis with predominance of microecological disorders of II (41.7%) and III (37.3%) degrees was detected in all children receiving anti-tuberculosis therapy. The microbial landscape of the intestine was characterized by a decrease in the quantitative content of representatives of the obligate flora Bifidobacterium spp. in 66.7% of children and Lactobacillus spp. in 45.8%, an increase in the number of facultative microorganisms: E. coli lac+ and Citrobacter freundii in 12.5%. Maintaining a full-fledged and active intestinal normobiota is possible by ensuring its dynamic control in children with tuberculosis during treatment. The results of studying the characteristics of the microbiota suggest that patients need probiotic therapy during the treatment of tuberculosis. Conclusion. Microecological disorders in children with tuberculosis therapy were characterized by a decrease in the quantitative content of representatives of obligate flora and an increase in the number of facultative microorganisms. Also in the material, various types of fungi of the genus Candida were identified.
Aim. To defne the role of Enterococcus spp. in the gut microbiota ensemble in patients with pulmonary tuberculosis. Materials andMethods. Inoculation of intestinal cultures was performed in 64 patients with multidrug-resistant tuberculosis before the specific therapy (30 patients and 55 strains) and after 1 month of the treatment (34 patients and 63 strains). We then assessed the production of haemolysins, gelatinase, phospholipase, adhesion factors, and organic acids.Results. A high prevalence of intestinal dysbiosis (66.7%) were detected in patients before the start of anti-tuberculosis therapy. Specifc treatment was associated with a decrease in Lactobacillus spp. along with an increase in C. perfringens and Candida spp. titers as well as with higher prevalence of E. coli lac. Before the treatment, Enterococcus spp. showed pronounced adhesion properties, providing colonization of the intestinal mucosa in titers of 6 (4; 7) lg CFU/g, although not producing invasion enzymes. Upon the 1 month of anti-tuberculosis treatment, adhesion capability reduced and 36% of the strains were low-adhesive. An increase of lipase-producing strains to 18% suggested the adaptation of Enterococcus spp. to a high content of lipids and fatty acids in the intestine.Conclusion. In patients with pulmonary tuberculosis having high prevalence of intestinal dysbiosis, Enterococcus spp. are frequent symbionts well integrated into the gut microbiota.
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