Analyses of the Effects of Arginine, Nicotine, Serotype and Collagen-Binding Proteins on Biofilm Development by 33 Strains of Streptococcus mutans

Streptococcus mutans serotype k strains comprise <3% of oral isolates of S. mutans but are prominent in diseased cardiovascular (CV) tissue. Collagen binding protein (CBP) genes, cbm and cnm, are prevalent in serotype k strains and are associated with endothelial cell invasion. Nicotine increases biofilm formation by serotype c strains of S. mutans, but its effects on serotype k strains and strains with CBP are unknown. Saliva contains arginine which alters certain properties of the extracellular polysaccharides (EPS) in S. mutans biofilm. We examined whether nicotine and arginine affect sucrose-induced biofilm of S. mutans serotypes k (n = 23) and c (n = 10) strains with and without CBP genes. Biofilm mass, metabolism, bacterial proliferation, and EPS production were assessed. Nicotine increased biomass and metabolic activity (p < 0.0001); arginine alone had no effect. The presence of a CBP gene (either cbm or cnm) had a significant effect on biofilm production, but serotype did not. Nicotine increased bacterial proliferation and the effect was greater in CBP + strains compared to strains lacking CBP genes. Addition of arginine with nicotine decreased both bacterial mass and EPS compared to biofilm grown in nicotine alone. EPS production was greater in cnm + than cbm + strains (p < 0.0001). Given the findings of S. mutans in diseased CV tissue, a nicotine induced increase in biofilm production by CBP + strains may be a key link between tobacco use and CV diseases.

Influence of siderophores and iron on Mycobacterium bovis isolation from pathological material

The article presents the results of studying the effect of siderophores and iron on the isolation of Mycobacterium bovis from pathological material. It has been established that the simultaneous presence of iron and siderophore from M. phlei in the nutrient medium makes it possible to detect the growth of M. bovis from pathological material 6–8 days earlier; ensures the growth of more colonies and bacterial mass. The presence of heterologous to mycobacteria siderophore (from Nocardia spp.) in the medium reduces the elective (growth) properties of the medium. Siderophores found in the culture filtrate or alcoholic extract of M. phlei can be valuable additives to culture media for the accelerated isolation of M. bovis from pathological material

Comparative analysis of the detectability of microorganisms in the prostatic fluid and ejaculate according to the results of bacteriological analysis

Introduction. Currently the chronic prostatitis (NIH type IV) remains insufciently studied and difcult to treat with antibiotics. When making the diferential diagnosis of chronic prostatitis it is generally accepted by the EAU and Russian guidelines that the detected microorganisms in the prostatic fuid and the ejaculate practically do not difer from each other. The tactics of treating patients with asymptomatic prostatitis by means of antibacterial drugs remains disputable. Most reputable authors believe that this category of patients shall receive treatment in case of infertility, pregnancy miscarriage or forthcoming surgery on the prostate gland. Recently, EAU guidelines have expressed doubts regarding the identity of the microbiota found in the prostatic secretions and the ejaculate.The study objective. Comparative analysis of the detectability of microorganisms in prostatic secretions and the ejaculate by means of bacteriological analysis among the men with chronic prostatitis of category IV.Materials and methods. The study involved 117 men who went to the urological clinic to pregravid examination or on the occasion of infertility. All patients were diagnosed with prostatitis of category IV based on a standard examination, and then a bacteriological analysis of prostatic secretions and ejaculate was performed. During the research the prostatic fuid and ejaculate were taken simultaneously and the analysis was carried out in one and the same bacteriological laboratory. In case the titer of the detected microorganisms was signifcant, the patients were ofered to undergo treatment without the use of antibiotics but with combination of bovhyaluronidase azoximer with prostate protectors and/or OM-89; after that a control bacteriological study was performed.Results. In the prostatic secretions and in the semen, the most commonly found gram-positive microorganisms were Staphylococcus spp. and Enterococcus spp.; the representatives of the Enterobacteriaceae were found less frequently. It was statistically proven that the titer of bacteria in the prostatic secretions was considerably higher than that in the ejaculate (p <0.01). Sterile cultures were statistically-proven to be more frequent in ejaculate compared with prostatic secretions (13.7 % vs 3.4 %, p <0.01). When analyzing the coincidences for bacteria, low concordance was obtained for all identifed microorganisms (gram-positive bacteria varied from 48.3 to 79.5 %, gram-negative bacteria varied from 57.1 to 80.0 %). After the combined therapy, the bacterial mass signifcantly decreased in the prostate secretion from 104.3 ± 1.6 CFU/ml to 103.3 ± 2.0 CFU/ml (p = 0.008), while in the semen the bacterial mass decreased from 103.5 ± 1.8 CFU/ml to 102.6 ± 2.1 CFU/ml (p = 0.02). In the prostatic secretions, there was a statistically-proven decrease in the number of gram-negative bacteria (p = 0.05). As soon as the treatment was completed all the patients demonstrated the normalized number of leukocytes according to microscopy of prostate secretions or spermogram.Conclusion. The ejaculate is a more sterile biomaterial compared with the prostatic fuid, which should be taken into account in the diferential diagnostics of the chronic prostatitis of category IV and MAGI. The use of non-antibacterial treatment regimens, such as bovhyaluronidase azoximer, prostate protectors and OM-89, can reduce the titer of bacteria to insignifcant values both in the prostatic secretions and in the semen; and in some cases make the ejaculate sterile.

Biofilm Early Stages of Growth and Accumulation Theoretical Model

A theoretical model to translate the evolution over time, in early stages, of growth and accumulation of biofilm bacterial mass is introduced. The model implies the solution of a system of differential-difference master equations. The application of an algorithm like Miller&acute;s tree term recurrence, already known for Bessel functions of first kind, allows an exact calculation of the solutions of such equations, for a wide range of parameters values and time. For biofilm model a five term recurrence is deduced and applied in a backwards computation. A suitable normalisation condition completes the reach of the solution.

Development and manufacture of erythrocyte pullorosis diagnosticum for the diagnosis of avian pullorosis typhoid

The article is devoted to the problems of diagnostics of poultry typhoid fever and the development of a technology for the manufacture of diagnosticum against this disease. A method of making erythrocyte diagnosticum for the diagnosis of poultry typhoid fever is shown, including obtaining the bacterial mass of Salmonella pullorum-gallinarum, isolating the antigenic fraction from it by treating the bacterial mass with a surfactant with the addition of soda or alkali in distilled water at 93-96 °C, followed by sensitization of formalinized erythrocytes, their purification and obtaining the target product in the form of 10% suspension, characterized in that 1-1.5% aqueous solution of Desmol is used as a surfactant to isolate the antigenic fraction, taken in a final weight concentration of 0, 1-0.3%, and the sensitization of formalinized erythrocytes is carried out in the presence of the sodium salt of chitosan succinate taken in a final weight concentration of 0.5-1.5%. In the industrial production of the diagnosticum at the initial stage, the bacterial suspension was necessarily mixed and the optical concentration was measured photometrically. The concentration of the surfactant solution was adjusted to 25 ml of microbial cells in 1 ml. At the second stage, antigen-sensitin was obtained and stored at 4 ° C. Sheep erythrocytes were used for sensitization. At the third stage, 20% formalized ram erythrocytes were obtained. Formalized erythrocytes were washed five times until the supernatant was completely cleared and sensitized. At the fourth stage, 300-500 ml was added to 1 liter of erythrocyte suspension for sensitization. sensitin and kept in a water bath at a temperature of 600С. At the fifth stage, the sensitized erythrocytes were washed to remove residual sensitin not associated with erythrocytes. Obtaining highly effective erythrocyte diagnostics for the diagnosis of avian pullorosis typhoid is an urgent problem. We have produced, improved and optimized the technology of industrial production of erythrocyte pullor antigen from the Salmonella pullorum-gallinarum strain for the diagnosis of avian pullorosis-typhus. We have theoretically substantiated and tested in production conditions a new method of resuspension, extraction, clarification of the bacterial mass. Used surfactants (surfactants) to obtain sensitin.

Features of amniotic fluid microbiota in full-term pregnancy

Objective. To study the microbial landscape of amniotic fluid in physiological process of full-term pregnancy. Recently, after publication of a number of studies regarding human microbiota (The Human Microbiome Project HMP), there occurred a change in paradigm on absolute sterility of fetal membranes and amniotic fluid in physiologically developing pregnancy. Materials and methods. At the City Clinical Hospital № 1 named after N.I. Pirogov, during elective cesarean section of 19 pregnant women (at the terms of 3741 weeks) with intact fetal membranes, an amniotic fluid of the following microorganisms was taken by means of PCR-PB: Lactobacillus spp., Enterobacteriaceae, Streptococcus spp., Staphylococcus spp., Gardnerella vaginalis / Prevotella bivia / Porphyromonas spp., Eubacterium spp., Sneathia spp. / Leptotrihia spp. / Fusobacterium spp., Megasphaera spp. / Veillonella spp. / Dialister spp., Lachnobacterium spp. / Clostridium spp., Mobiluncus spp. / Corynebacterium spp., Peptostreptococcus spp., Atopobium vaginae, Mycoplasma hominis, Ureaplasma (urealyticum + parvum), Candida spp., Mycoplasma henitalium. Results. The general bacterial mass (GBM) of amniotic fluid in intact fetal membranes is 103,02 Ge/copies, in 47.4 % of cases the amniotic fluid is sterile. Microbiota is most often presented by Enterobacteriaceae spp. 37 %, the share of the rest, identified bacteria is 28 %, the share of unknown is 35 %. Conclusions. In case of physiologically developing pregnancy and intact fetal membranes, the general bacterial mass is low (GBM = 103,02 345 Ge/ml). In the intact amniotic sac the most typical microorganisms living in amniotic fluid are Enterobacteriaceae spp. (37 %), the rest are presented in single instances. The presence of the representatives of anaerobic vaginal dysbiosis as well as lactobacilli is not typical for the intact fetal membranes.

INFLUENCE OF MICROBIOTA ON THE CLINICAL AND IMMUNOHISTOCHEMICAL CHARACTERISTICS OF CHRONIC GENERALIZED CATARRHAL GINGIVITIS IN CHILDREN

The aim: The purpose of the study is to characterize the influence of quantitative and qualitative composition of gingival microbiota on the status of the main immune system cells, localized in the gums, in chronic generalized catarrhal gingivitis in children. Materials and methods: The study involved 26 children aged 9 to 16 years, patients with chronic generalized catarrhal gingivitis mild to moderate severity (CGCG) and 18 children with intact gums were comparison group. We determined the hygienic indices Fedorov, has been received, Silness-Loe, PMA, bleeding index for Myuleman and intensity of caries index CFD + cf, CFD. Histological and immunohistochemical studies were performed on serial sections kriostatnyh who made biopsy of gingival papillae. Microbiological study gingival part of crown plaque was performed by multiplexed PCR in real time. Results: Value hygienic indices in children with CGCG higher than in healthy, indicating the difficulty of care in the presence of periodontal inflammation. As a result of immunohistochemical studies revealed that HLA-DR + cells under conditions of active disease migrate to mucosal lamina propria epithelium. Number of CD3 + cells in the epithelium CGCG was significantly higher than the number in the intact epithelium and was the most numerous of population. In the biopsy of affected children significantly reduced the number of CD4 + cells. When CGCG quantitative total bacterial mass, Lactobacillus spp., Enterobacteriaceae, Gardnerella vaginalis / Prevotella bivia / Porphyromonas spp. in the sample CROWN dental plaque was significantly higher than rates under physiological conditions, and may serve as diagnostic criteria of dysbiosis. Conclusions: So, CGCG is a disease in the etiology of which is one of the leading roles played by microbial factor, namely, the value of the quantitative ratios of certain types of microorganisms of dental plaque compared to the total bacterial mass of plaque. Therefore, it is reasonable to include comprehensive treatment CGCG drugs in children, leading to natural immunostimulation which causes restoration of local immunity in the gum tissue and drugs to restore quantitative and qualitative composition of normal microflora of the child, thus providing a high therapeutic effect and serve as justification their choice.

Biofilm Early Stages Growth and Accumulation Theoretical Model

A theoretical model to translate the evolution over time, in early stages, of growth and accumulation of biofilm bacterial mass is introduced. The model implies the solution of a system of differential-difference master equations. The application of an algorithm like Miller&acute;s tree term recurrence, already known for Bessel functions of first kind, allows an exact calculation of the solutions of such equations, for a wide range of parameters values and time. For biofilm model a five term recurrence is deduced and applied in a backwards computation. A suitable normalisation condition completes the reach of the solution.