Биологические свойства грамнегативных бактерий, верифицированных в моче пациенток с рецидивирующей неосложненной инфекцией нижних мочевых путей

Uncomplicated infections of the lower urinary tract are more often recorded in women and are characterized by recurrent severe course. The lack of effective approaches to the management of this cohort of patients and the insufficient knowledge of the qualitative characteristics of uropathogens hinder the successful treatment of these infections. The purpose of this project is to study the biological properties of representatives of Gram-negative bacteria isolated from the urine of patients with recurrent uncomplicated lower urinary tract infection at various levels of bacteriuria.We present here the results of the one-stage cohort study of 62 patients 20-50 years old with recurrent uncomplicated lower urinary tract infection. The level of bacteriuria, genes of virulence factors, hemolytic, adhesive and anti-lysozyme activities were determined for the Gram-negative bacteria isolated from the urine of patients.Representatives of Gram-negative microorganisms (62 strains) were isolated from the urine of all patients. Their bacteriuria levels ranged from 102 to 108 CFU/ml. A wide range of the virulence factor genes with the prevalence of papGII, feoB, fyuA, were found in all strains regardless of the degree of bacteriuria. Twenty significant (p<0.05) coefficients of mutual contingency between different pairs of genes were found. We found that the hemolytic activity of Enterobacteriaceae is statistically significantly associated with papA, feoB, fyuA, kpsMII, and usp, while the anti-lysozyme activity is associated with papA, afa, and usp.The detection of genotypic and some phenotypic traits expressed in various variations in all Gram-negative bacteria regardless of their bacteriuria levels creates a potential risk for recurrent of the lower urinary tract infection. Consequently, even the minimal levels of bacteriuria become clinically significant. Therefore, in bacteriological diagnostics it is necessary to determine all levels of bacteriuria, including the minimum level (102 CFU/ml), since isolated microorganisms at this level have not only adhesive and anti-lysozyme activities, but also contain genes encoding the virulence factors. Неосложненные инфекции нижних мочевых путей чаще регистрируются у женщин и характеризуются рецидивирующим тяжелым течением. Отсутствие эффективных подходов по ведению данной когорты пациенток и недостаточная изученность качественных характеристик уропатогенов препятствуют успешному лечению этих инфекций. Цель настоящего исследования — изучить биологические свойства представителей грамнегативной микробиоты, выделенной из мочи паци­енток с рецидивирующей неосложненной инфекцией нижних мочевых путей, при различных уровнях бактериурии.Проведено когортное одномоментное исследование 62 пациенток в возрасте 20–50 лет с рецидивирующей неосложнен­ной инфекцией нижних мочевых путей. У грамнегативных бактерий, выделенных из мочи пациенток, определяли уровень бактериурии, гены факторов вирулентности, гемолитическую, адгезивную и антилизоцимную активности.У всех пациенток из мочи выделены представители грамнегативных микроорганизмов (62 штамма). Уровни бактери­урии колебались от 102 до 108 КОЕ/мл. У всех штаммов обнаружены гены факторов вирулентности в широком диапазоне с превалированием papGII, feoB, fyuA независимо от степени бактериурии. Обнаружено 20 достоверных (p<0.05) коэффи­циентов взаимной сопряженности между различными парами генов. Гемолитическая активность энтеробактерий значимо связана с papA, feoB, fyuA, kpsMII и usp, антилизоцимная – с papA, afa и usp.Таким образом, наличие генотипических и некоторых фенотипических признаков в различных вариациях экспрессии у всех представителей грамнегативных бактерий при всех уровнях бактериурии создает потенциальный риск развития очередного рецидива инфекции нижних мочевых путей. Поэтому даже минимальные уровни бактериурии являются кли­нически значимыми. Таким образом, при бактериологической диагностике необходимо определять все уровни бактери­урии, включая минимальный (102 КОЕ/мл), так как выделенные микроорганизмы при данном уровне обладают не только адгезивной и антилизоцимной активностями, но и имеют гены факторов вирулентности

Urine and colon microbiota in patients with recurrent uncomplicated lower urinary tract infection

Background. The prevalence of urinary tract infections (UTIs) in general and recurrent lower UTIs, in particular, constantly remains at a high level. Currently, more and more researchers are attempting to study the microbial biocenosis of the urinary tract in patients with UTIs in terms of the relationship between UTIs and patient's colon microbiota in order to detail the etiology and pathogenesis of these conditions as an insufficiently studied issue.Objectives. To study the microbiota of urine and colon, as well as significant correlations in these biotopes.Materials and methods. 169 patients with recurrent lower UTIs from the Urology Clinic and the Department of Microbiology and Virology No. 1 (Rostov State Medical University) were examined. Average age: 36.2 ± 4.7 years. Midstream urine samples for bacteriological examination was taken in accordance with the rules set out in the Clinical recommendations (2014) exactly observing the preanalytical stage. The study of the colon microbiota as well as the collection of material was carried out in accordance with the rules set forth in the industry standard (OS) OST 915000. 11.0004 - 2003, Order of the Ministry of Health of the Russian Federation No. 231 of 09/09/03. Guided by this standard, the interpretation of the results was conducted. Results' statistical processing was implemented using the SPSS version 23 package.Results. In patients with recurrent lower UTIs two significant quantitative correlation coefficients were detected. The first correlation coefficient showed inverse correlation between typical E. coli isolated from urine and Eubacterium spp. isolated from feces (r = -0.434, p = 0.009). The second correlation coefficient showed a direct correlation between the amount of Lactobacilli isolated from urine and Coalulase-negative Staphylococci (CoNS) isolated from feces, accordingly (r = -0.434, p = 0.009).When comparing the frequencies of detection of various microbiota taxa, three significant correlation coefficient were found in the loci under study: between Lactobacilli or Peptococci isolated from urine and CoNS verified in feces (PCC - 0.342, p = 0.031 and PCC - 0.341, p = 0.018, respectively), as well as between Peptococci detected in urine and Enterococci in feces (PCC - 0.349, p = 0.028).Conclusion. The data we obtained on significant correlation coefficients between microorganisms isolated from urine and colon indirectly confirm the translocation mechanism. Undoubtedly, further research is needed to study the mechanism of translocation of microorganisms from the intestines and other biotopes into the organs of the urinary system in order to understand the pathogenesis of this common group of diseases

Is there a relationship between the urine, vagina, and gut microbiota in patients with an infection of the upper urinary tract?

Introduction. Urinary tract infections of any localization are rarely considered in the context of the endogenous relationship of patient's urine microbiota and the microbiota of nearby biotopes. Nowadays, there are attempts to study the microbial interrelationships of the urinary system with nearby biotope to find fundamentally new solutions in the study of the etiology and pathogenesis of diseases, but now this problem remains almost unexplored.Purpose of the study. Analysis of the correlations between various taxa of the microbiota isolated from urine, vagina and intestines in patients with upper urinary tract infection.Materials and methods. According to the inclusion criteria, bacteriological examination of cystic urine, feces and posterior vaginal fornix discharge was performed on 60 women (18-65 years old) with acute obstructive pyelonephritis. The material was taken, transported and examined by standard methods, but with some modifications of nutrient media. Statistical calculations were performed in R "R ver 3.2" ("R Foundation for Statistical Computing", Vienna, Austria) with generally accepted significance coefficients.Results. 26 significant correlation coefficients were found in the urinary tract and the vagina when conducting a comparative correlation analysis between the microbiota taxa, and 21 in the urinary tract and intestine. Significant correlations between the general taxa were direct and a large proportion of them were in optional anaerobic microbiota. 28 significant correlation coefficients were found in most cases between aerobic and anaerobic taxa of microorganisms in the vagina and intestines.Conclusion. The obtained significant correlation coefficients between various microbiota taxa in the three studied biotopes are evidence of the relationship of these loci. However, further research is needed on the phylotyping and genotyping of the microbiota taxa of the urinary tract, vagina and intestines of patients with urinary tract infection

Antimicrobial resistance of uropathogens in patients with acute obstructive pyelonephritis

RELEVANCE: Acute pyelonephritis is known to be the most complicated and severe urinary tract infection occurring in all age groups and accounting for 14% of all kidney diseases. The generally recognized standard antibiotic therapy cannot completely prevent the progression of the disease to its chronic form after relief of its acute manifestations thus leading to a high incidence of relapses. The aim of our study was to investigate the spectrum of uropathogens and their antibiotic sensitivity in acute obstructive pyelonephritis.MATERIALS AND METHODS: The study comprised 72 patients who underwent semi-rigid ureteroscopy and ultrasonic lithotripsy for ureteral stones. In all patients, bladder urine samples collected by a transurethral catheter were tested bacteriologically using an extended set of culture media within 3 hours after hospital admission. Antibiotics used in antibiotic sensitivity testing for all uropathogens, were grouped into 4 classes (carbapenems, fluoroquinolones, cephalosporins, penicillins). Etiotropic treatment was started upon the availability of the spectrum of microbial patterns, the level of bacteriuria and antibioticogram of uropathogens, 5-6 days after administering initial empirical antibiotic therapy.RESULTS: The study patients had a high detection rate (83.3%) of canonical uropathogens in the bladder urine identified due to using an extended set of culture media, with a bacteriuria of more or equal 103 CFU/mL. Given the results of local antibiograms, a rational antimicrobial therapy should include carbapenems, namely ertapenem or meropenem as initial empirical antibiotics. Using fluoroquinolones as the first line treatment can lead to an inadequate effect in 15.0 to 67.0% of the cases. The findings of the antibiotic resistance testing of uropathogens to cephalosporins and semisynthetic penicillins showed that they should not be used as initial empirical antibiotic therapy for acute obstructive pyelonephritis in the given department of urology

The gene profile of Enterobacteriaceae virulence factors in relation to bacteriuria levels between the acute episodes of recurrent uncomplicated lower urinary tract infection

© 2020 Informa UK Limited, trading as Taylor & Francis Group. Objectives: The pathogenic potential of uropathogens isolated between acute episodes of recurrent lower urinary tract infection (rLUTI) is studied insufficiently. The objectives were to determine the spectrum of virulence genes of Enterobacteriaceae cultured between acute episodes of rLUTI at various levels of bacteriuria. Methods: Bacteriological examinations of 169 premenopausal women’s midstream urine with rLUTI were performed between acute episodes of UTI. Sixty-two strains of Enterobacteriaceae at concentrations 102–108 CFU/ml were analyzed for the presence of papA, papE/F, papGII, afa, bmaE, iutA, feoB, fyuA, kpsMTII, and usp virulence factors genes’ (VFGs) fragments. Results: In all strains VFGs were found with numbers from 1 to 10. Four VFGs were found at all levels of bacteriuria (from 102 to 107−8) in most strains (\u3e50%): papGII, feoB, fyuA, usp. In total, 28 significant Pearson contingency coefficient (PCC) were determined. Each of the genes, papA, papE/F, usp, was found more often in uropathogens from patients with a higher level of leukocyturia. Conclusions: The inter-episode period in rLUTI is associated with varying levels of bacteriuria of enterobacteria. Since enterobacteria virulent potential could be determined at all levels of bacteriuria, there is at all levels of bacteriuria a potential risk for recurrence of LUTI