[Taxonomic characteristics and antibiotic susceptibility of microorganisms verified in the expressed prostate secretion in post-COVID-19 patients with recurrent chronic bacterial prostatitis].

INTRODUCTION: Recurrent chronic bacterial prostatitis (rCBP) is a hard-to diagnosis-and-treat disease which there is no consensus. A particularly difficult cohort is represented by patients who had COVID-19. The study aimed to evaluate the taxonomic structure and sensitivity to antibacterial drugs of microorganisms verified in expressed prostate secretion (EPS) in rCBP-patients who had COVID-19. MATERIALS AND METHODS: A multicenter, prospective, randomized study was conducted with the inclusion of 52 rCBP patients who had COVID 19, in which the taxonomic structure and susceptibility were studied to antibacterial drugs of microorganisms that were verified and dominated in the EPS. Bacteriological study was carried out using an extended set of selective nutrient media and special cultivation conditions. Antibiotic susceptibility was determined in the taxa of microbiota dominating in the EPS. RESULTS: The mean age of the patients was 34.8+/-5.2 years, the duration of rCBP was 5.7+/-2.3 years. In all patients, various variants of aerobic-anaerobic compositions of microorganisms were recorded in the life cycle. A total of 27 microbiota taxa were isolated. The aerobic cluster was represented by 16 genera and/or species, the anaerobic cluster by 11. When studying antibiotic susceptibility to antibacterial drugs, an increase in antibiotic resistance of the most microorganisms isolated was revealed. CONCLUSIONS: The taxonomic structure of microorganisms in rCBP-patients who had COVID-19 in all cases was characterized by complex and new variants of aerobic-anaerobic associations of microorganisms. When studying the antibiotic susceptibility, multi-resistant and pan-resistant bacteria were identified that is a real threat to this category of patients.

[Extended culture study as a keypoint to rethinking antibiotic therapy for chronic bacterial prostatitis].

OBJECTIVE: To juxtapose the microbiological efficacy of standard and targeted antibiotic therapy (ABT) based on the comparison of the results of extended bacteriology of biomaterial in patients suffering chronic bacterial prostatitis (CBP) before and after treatment. STUDY DESIGN: single-centre observational comparative study. Sixty patients with CBP aged 20 to 45 years were included in the study. All patients underwent an initial examination: questioning, Meares-Stamey 4-glass test, extended bacteriology of biomaterial samples, and determination of antibacterial susceptibility (ABS). After the initial examination, the patients were randomly assigned to two groups (30/30 patients). In group (G) 1, antibacterial drugs were prescribed following the EAU guidelines on Urological Infections (monotherapy), in G2, focusing on the results of ABS (mono or combination therapy). Evaluation of the treatment effectiveness and control bacteriology were carried out three months after therapy. RESULTS: In G1 vs G2, nine vs ten aerobes and eight vs nine anaerobes were identified in the expressed prostate secretion, respectively. The microbial load of the samples in more or equal 103 CFU / ml was established in G1 vs G2 for five vs ten aerobes and seven vs eight anaerobes, respectively. The highest ABS of bacteria was determined to moxifloxacin, ofloxacin, and levofloxacin. Cefixime was the most active against anaerobes. After treatment, no significant changes in the bacterial spectrum were observed in both groups. A more reliable decrease in the frequency of microorganism identification and the microbial load of the samples was observed in patients with G2 after the targeted ABT. CONCLUSION: Targeted ABT based on extended bacteriology can be considered an effective alternative to standard guideline-approved ABT for the treatment of CBP.

[Cytological examination of urine in the differential diagnosis of recurrent lower urinary tract infection].

INTRODUCTION: A diagnosis and treatment of recurrent lower urinary tract infection (UTI) in women is still one of the unresolved problems of urology. The proper identification of the etiological factor determines the treatment tactics. Therefore, the most urgent issue of recurrent lower UTI is the differential diagnosis of the causative pathogens. MATERIALS AND METHODS: A cytological study of urine was performed in 151 patients with recurrent lower UTI, who, according to the results of bacteriological and PCR studies of urine, were divided into three groups, depending on the etiological factor. The group 1 (n=70) included women with recurrent lower UTI of bacterial etiology, while in group 2 (n= 70) papillomavirus etiology was found, and in group 3 (n=11) candida species were the causative pathogens. The age of the patients ranged from 20 to 45 years (32.3+/-7.8). RESULTS: In the majority of patients with recurrent lower UTI of bacterial etiology, the cytological features were represented by leukocytes, plasma, epithelial cells and bacteria in combination with actively phagocytic macrophages. In group 3, in addition to a large number of leukocytes (neutrophils) and epithelial cells, Candida mycelium was found. In group 2, signs of bacterial inflammation were minimal, while a predominance of lymphocytes, epithelial cells, and the presence of single neutrophils was seen. With papillomavirus lesions of the bladder, urothelial cell dystrophy with the presence of koilocytes developed. CONCLUSIONS: A cytological examination of urine can confirm the etiology of the recurrent lower UTI and be an evidence-based criterion in the differential diagnosis of bacterial, candidiasis, and papillomavirus infection. Total transformation of the urothelium and vacuolization of urothelial cells, as well as an excess of lymphocytes in the urine in the absence of neutrophils, are the characteristic features of viral recurrent lower UTI.

[Long-term inflammatory and neoplastic reaction of prostate tissues during its transurethral infection with uropathogens: evaluation of the results of animal model study].

INTRODUCTION: There is no convincing evidence of the persistence of acute or the development of chronic bacterial-induced prostatic inflammation in the long term when infected with various titers of the uropathogen. Along with this, controversial data are presented on the relationship between post-infectious chronic inflammation and neoplastic changes in prostate tissues. OBJECTIVE: To carry out, based on the experimental data: 1) assessment of the degree of bacterial contamination and the severity of histological changes in prostate tissues on the 60th follow-up day in case of transurethral infection with various uropathogens in titers of 102,3,5 CFU/ml; 2) fundamental comparative analysis between the indicators of the inoculated test-titer and microbial load with the severity of histological changes in prostate tissues; 3) verification of neoplastic transformations in the prostate tissues during a long-term persistent bacterial-induced inflammatory process. MATERIALS AND METHODS: Animal studies were conducted using FELASA protocols. Laboratory animals: 14 New Zealand rabbits. Tested uropathogens: aerobes - E. coli, S. haemolyticus, anaerobes - P. niger. Titers: 102,3,5 CFU/ml. Uropathogen inoculation technique: topical transurethral. RANDOMIZATION: all laboratory animals were divided into 5 groups according to the uropathogen (4 experimental, 1 control). Follow-up period: 60 days. Sacrification and autopsy of the animals were performed on day 60. Biopsies were taken from various parts of the prostate, as well as from the bladder neck and the edge of the membranous urethra. Cultural, histological and immunohistochemical (expression of p53 and Ki-67) studies of prostate tissues were conducted. Statistical data processing was performed using the GraphPad Prism 9.0 program (GraphPad Software Inc., Graphpad Holdings LLC, San Diego, CA, USA) applying descriptive and non-parametric statistics. RESULTS: Two individuals infected with S. haemolyticus + P. niger had a lethal outcome. The contamination of prostate tissue was determined in all cases of infection. In 88.9% of the cases, an increase in tissue microbial load was determined compared to the initial titer. Multivariate analysis of culture study values revealed the presence of intragroup differences in prostate contamination only between infection with E. coli 103 CFU/ml and E. coli 105 CFU/ml (p=0.006), as well as intergroup differences between infection with E. coli 105 CFU/ml and P. niger 105 CFU/ml (p=0.013). The histological study revealed moderate proliferative inflammation after inoculation with 102,3,5 CFU/ml in the E. coli and S. haemolyticus groups. In the case of S. haemolyticus, it was more pronounced due to the presence of persistent alterative lesion foci; in the P. niger group, mild proliferative transformations were observed in prostate tissues in all cases. The immunohistochemical study of changes determined p53 expression (10.0%) in some areas of the glandular epithelium of prostate glands (but without a positive internal control) only in case of infection with E. coli 105 CFU/ml. Areas of glandular epithelium with Ki-67 expression ( less or equal 25.0%) were visualized in all tested groups, mainly at titers of 103 and 105 CFU/ml, but the severity of proliferative activity was not high (1+). There were no foci of prostate tissue with simultaneous nuclear activity of p53 and Ki-67. CONCLUSION: Proliferative inflammation of different intensity in prostate tissues was observed after sixty days. Its severity was mainly determined by the type of infecting agent (S. haemolyticus > E. coli > P. niger) and was not dependent on the inoculated titer and the subsequent microbial load of prostate tissues. No areas of neoplastic transformation of prostate tissues were reliably identified in the case of a bacterial-induced inflammatory process in the estimated follow-up period.

[Asymptomatic bacteriuria in pregnant women the normal condition of healthy women urine].

OBJECTIVE: to study the dynamics of the microbiota of a freshly excreted middle portion of urine in primigravida at different times of all three trimesters of pregnancy. MATERIALS AND METHODS: A single-center prospective observational cohort study was conducted with a consecutive enrollment of 30 women at different gestational ages: I - 8-12 weeks, II - 22-24 weeks, III - 30-32 weeks. A midstream specimen of morning vesical urine was taken for the study, then it was cultivated using nutrient media for aerobic and anaerobic microorganisms under appropriate conditions. Statistical analysis was performed using SPSS ver.26 (IBM SPSS Inc., Chicago, IL, USA). RESULTS: The freshly released middle portion of urine in all 30 observations in the I-III trimesters contains aerobic-anaerobic associations of microorganisms. Coagulase-negative staphylococci, Enterococcus spp., Corynebacterium spp., Lactobacillus spp., Eubacterium spp. prevail in the urine during pregnancy. The E. coli, Candida spp. detection frequency decreases by the third trimester, but Lactobacillus spp. detection frequency rises. Significant differences in the detection frequency were found only in Propionibacterium spp. and Lactobacillus spp. The average level of bacteriuria in most cases is 102-103 CFU/ml with significant differences only in E. faecium, Lactobacillus, Propionibacterium spp. in the III trimester. DISCUSSION: The study of urine at different times of all three trimesters of pregnancy refutes the previous ideas about asymptomatic bacteriuria. The urine microbiota in primigravida during pregnancy has wide spectrum and quite stable until delivery. Such bacteriuria can be considered asymptomatic, but it is a consequence of a healthy state and it is not a disease or its predictor. CONCLUSION: The term asymptomatic bacteriuria is not correct in the context of risk factor of urinary tract infection in pregnant women.

[Virusuria as a component of the urine microbiota and its significance for assessing the health of the urinary tract: a descriptive clinical study].

AIM: To determine the frequency of occurrence of oportunistic pathogenic bacterial flora and viral pathogens in the urine of healthy people with the establishment of the association between them. MATERIALS AND METHODS: 40 healthy sexually active women and men were examined, which are divided by gender into equivalent groups: Group I - healthy women (n=19), Group II - healthy men (n=21). The age of the subjects ranged from 20 to 25 years, the average age was 22.4+/-1.2 years. In both groups, the average portion of morning urine was taken for a study after a proper hygienic procedure with self-urination of the subjects in a sterile plastic container (Sterile Uricol for urine sample collection "HiMedia"). In addition to the nutrient media regulated by the Clinical Guidelines, additional HiMedia chromogenic media were used to cultivate facultative anaerobic (FAB) and non-clostridial anaerobic bacteria (NAB). Detection of viruses was performed by PCR with detection in "real time". DNA isolation was carried out by the sorption method using the AmpliPrime DNA-Sorb-B ("NextBio") kit from urine samples, with preliminary concentration. RESULTS: In all 40 cases, normative leukocyturia was detected in the urine. According to the results of bacteriological examination of urine, healthy men and women in all cases found aerobic-anaerobic associations. Coagulase-negative staphylococci (CNS) and Corynebacterium spp. Dominated in the cluster of aerobic taxa of microbiota. (75.0%, 55.0% respectively). The spectrum of CNS was represented by five species: S.epidermidis (30.0%), S.haemolyticus (27.5%), and S.warneri (25.0%), S.saprophyticus and S.lentus (15.0%). Enterococcus spp. were recorded in the urine in 32.5% of cases. Representatives of the Enterobacteriaceae family were represented by 4 taxa: E. coli (10.0%), Klebsiella spp., Proteus spp. (5.0% each), Enterobacter spp., Citrobacter spp. (2.5%). In a cluster of anaerobic bacteria in the urine, Eubacterium spp. (60.0%) and almost half of healthy individuals recorded Lactobacillus spp. and Peptococcus spp. (42.5% each). When analyzing the frequencies of detection of various microbiota taxa, it was found that women significantly more frequently recorded urine Corynebacterium spp., Eubacterium spp. and Lactobacillus spp., as well as Enterococcus spp. and Peptococcus spp. Peptostreptococcus spp. and Veillonella spp. were significantly more often determined (p<0.05) in the urine of men. HHV6 (10.0%), HPV18 and B19 parvovirus (2,5%) were determined in the urine of healthy people. It should be noted that the studied viruses were more often recorded in men, in particular, HPV18 and parvovirus B19 - only in men, and HHV6 more often in men (7.5%), less often in women (2.5%). Significant associations of some genera of microorganisms with the sex of the participants were revealed for E. faecalis and Lactobacillus spp., which were more often found in the urine of healthy women Reliably significant associations were found for three taxa: viruses HPV6, HPV18 and parvoviruses B19 (16.7%) were determined in the presence of Bacteroides spp., Bifidobacterium spp., and Prevotella spp., in urine. Accordingly, in 83.3% of cases, these viruses were detected in the absence of the above-listed taxa of microorganisms in the urine. CONCLUSIONS: The normal urinary microbiota of healthy women and men has differences: Lactobacillus spp and Candida spp are absent in the urine of men while Streptococcus spp in urine of women. HHV6, HPV18, parvoviruses B19 are found in urine of healthy people and more often in men. Data about the virobiota and microbiota of urine in healthy people can highlight on the pathogenesis of urinary tract infections of various localization and develop targeted approaches in personalized therapy of this group of diseases.

[Experimental evaluation of inflammatory process in the prostate caused by transurethral transmission of uropathogen in low counts].

BACKGROUND: According to the literature, bacterial count of uropathogens isolated from expressed prostate secretion and urine which is sufficient for a diagnosis of bacterial prostatitis I and II categories, remains contradictory. Undoubtedly, the identification of microorganisms from affected organ in high titers indicates the presence of a relevant infectious-inflammatory process. In turn, there is no consensus on the development of bacterial prostatitis at lower titers of uropathogens. Thus, the aim of our study was to identify and compare the potential features of the development and occurrence of an infectious inflammatory process in the prostate during the reproduction of bacterial prostatitis in an animal model using a low titer of causative uropathogens. MATERIALS AND METHODS: A total of 16 "New Zealand" mature male rabbits aged 24+/-2 weeks old with weight of 3.5+/-0.3 kg were examined. Inoculation was performed via transurethral route, according to the developed experimental technique. E. coli was used as bacterial agent with a count of 1 x 103 CFU/ml, 1 x 105 CFU/ml and 1 x 107 CFU/ml. All animals were randomized into 4 groups of 4 individuals depending on the titer of the inoculated microorganisms (groups 1-3, respectively), group 4 - control (with inoculation by Sol. NaCl 0.9%). Sacrification and vivisection were performed on days 1, 3, 7 and 14 of the control days. Biopsy specimens from the lower urinary tract and internal genital organs of laboratory animals (bladder, urethra, prostatic complex - 6 biopsies #1A-1D, 2A, 2B) were evaluated morphologically and bacteriologically. Analytical evaluation of the experimental data was presented using descriptive statistical methods. RESULTS: In experimental groups (Groups 1-3), bacteriological examination of prostatic complex biopsies showed growth of microflora in all samples in titers of 101-107 CFU / ml. In group 1, the maximum concentration of uropathogen was observed on day 7, compared to day 1 in both groups 2 and 3. In all observed cases, the highest degree of bacterial contamination was noted in the biopsy specimens from paraprostatic tissues and distal part of the prostate, which was 4.0+/-1.7 lg CFU/ml and 3.5+/-1.9 lg CFU/ml, respectively, and the smallest in proximal prostatic loci (1C) and bladder neck (2B) - 3.0+/-1.2 lg COE / ml and 3.0+/-1.7 lg COE / ml, respectively. According to the morphological study, a relevant progression of the suppurative and destructive inflammation (with foci of colliquation necrosis) was identified in group 1 in the biopsies from the prostate with a maximum degree of changes on day 7 with subsequent formation of loose connective tissue proliferation areas by 14 days. This indicates the conversion of the inflammatory process to the chronic stage. These changes corresponded with the results of histopathological studies in groups 2 and 3 where higher titers of bacterial agent were used. In group 4 (control) the commensal flora was bacteriologically determined in the biopsies, but there were no signs of inflammation, according to the results of the morphological study. CONCLUSION: In experimental model, we found that E. coli 103 CFU / ml induces the development of a phasic inflammatory process in the structures of the prostatic complex. These processes resulted in the formation of irreversible proliferative changes. As a consequence, it shold be recommended to consider these signs of contamination when evaluating the results of bacteriological examination of expressed prostate secretion/urine samples during planning treatment strategy.

[Bacterial prostatitis: epidemiology and etiology].

Prostatitis is considered as heterogeneous group of the diseases attracting broad interest of researchers worldwide. The acute and chronic stages of the process, as particular categories of bacterial prostatitis, remain in the realities of modern practical and scientific urology with lively discussions. Despite a large number of domestic and international publications, consensus on many topical issues of bacterial prostatitis is currently not achieved. The true incidence and prevalence of bacterial prostatitis in various regions of the world has not been finally determined. The exhaustive data concerning the etiological structure, mechanisms of development and persistence of the inflammatory process in the prostate tissue are not presented. At the same time, new risk factors that can influence the development and progress of these diseases are identified and discussed. Researchers review algorithms for examining patients, obligate spectrum of diagnostic methods, as well as the desirability of using the newest investment means necessary for the successful verification of the diagnosis taking into account the development of modern medical technologies. This review describes in detail the various aspects of the epidemiology and etiology of bacterial prostatitis, based on an analysis of major literature sources in conjunction with its own scientific facts.

[Long-term monitoring of leukocyturia and bacteriuria after acute obstructive pyelonephritis].

The generally accepted standard of antibiotic therapy aimed at causative agent does not exclude the risk of acute obstructive pyelonephritis (AOP) to become chronic after the end of antibiotic therapy, resulting in a high incidence of relapses. To assess the outcomes of the AOP, we undertook a prospective clinical trial. MATERIALS AND METHODS: The study comprised 51 consistently selected patients (12 men and 39 women) with AOP occurred as a result of unilateral ureteric calculus obstruction. Inclusion criteria for the study were as follows: no history of urological diseases, including urinary stones; the first presentation to the urologist with AOP; the level of the bladder bacteriuria before the drainage of the obstructed kidney more or equal 103 CFU/ml. Bladder urine samples were collected for bacteriological examination by catheterization. At one, 3 and 6 months after completion of the treatment of AOP, the patients underwent a general clinical examination, and midstream specimens of urine were collected from the patients for bacteriological analysis to control leukocyturia and bacteriuria. RESULTS: Eradication of causative uropathogens from urine occurred within 6 months in 96.1% of patients, and at that point, leukocyturia persisted in 23.5% of the patients. The decrease in detecting uropathogens in the urine was accompanied by an increase in the frequency of the presence in the urine of other aerobes and anaerobes, i.e., no sterile urine cultures were obtained from any patient. The rates of eradication of uropathogens were associated with the patients age, the duration of the pre-hospital stage of AOP, and the complicated course of AOP.

[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.

[Is bacteriological testing of bladder urine informative in acute obstructive pyelo- nephritis?]

The problem of the etiology and pathogenesis of acute obstructive pyelonephritis (OOP) remains one of the challenging issues of modern urology. Etiological agents of pyelonephritis can be both gram-negative and gram-positive opportunistic bacteria mostly belonging to the normal flora in humans. The generally accepted diagnostic work-up involves a bacteriological testing of not pelvic urine, but of bladder urine collected by a transurethral catheter or midstream specimens of urine collected from the patients. The aim of our study was to compare the microbiota of bladder and pelvic urine in patients with OOP. MATERIALS AND METHODS: The study comprised 72 sequentially selected patients (12 men and 60 women) with OOP associated with ureteral stones. Mean age of patients was 53.7+/-0.5 years. All patients underwent bacteriological examination of the bladder urine collected by a transurethral catheter and pelvic urine obtained after relieving stone-related ureteral obstruction. Urinary diversion was performed using j-j stent and PCN in 64 and 8 patients, respectively. Preoperative prophylactic antibiotics were administered routinely. Bacteriological testing of urine was carried out using an extended set (9-10) of culture media. Empirical antibiotic therapy was initiated only after the restoration of urine outflow from the kidney and continued for 5-6 days until the availability of bacteriological testing results. RESULTS: Levels of bacteriuria with Enterobacteria, gram-positive pathogens and NAB in two urine samples did not differ significantly (p>0.05). There was a wide range of bacteriuria from 101 to 106 CFU/ml of most microorganisms except @Proteus spp., S. aureus. In bladder urine, the rates of bacteriuria of more or equal 104 CFU/ml for E. coli, Klebsiella spp. and Proteus spp. were 90.9%, 72.7% and 100.0%, respectively. For the remaining microorganisms, predominant bacteriuria was less or equal 103 CFU/ml. In pelvic urine, the rates of bacteriuria of more or equal 104 CFU/ml for E. coli, Klebsiella spp. and Proteus spp. was 71.8%, 40.0% and 66.7%, respectively. Other uropathogens in the pelvic urine mainly had a bacterial count of less or equal 103 CFU/ml. Only the concentration of Corynebacterium spp. in the pelvic urine significantly (p=0.023) differed from that of the bladder urine. There were no significant differences between microbiota of bladder and pelvic urine depending on duration of OOP except higher rates of Corynebacterium spp. in the bladder urine.

[Microbiota of urine and vagina of healthy postmenopausal women (a pilot study)].

Studying microbiota of different urogenital tract habitats in healthy postmenopausal women is of practical importance in deciding on the appropriateness of correction of dysbiotic disorders. The aim of this study was to examine the vaginal and urine microbiota of healthy postmenopausal women. The study included 20 healthy postmenopausal women (mean age 59,0+/-2,1 years). Duration of menopause in all subjects was more than 8 years. Bacteriological testing of urine and vaginal specimen was carried out on the extended media (15) for cultivating facultative anaerobic bacteria (FAB) and nonclostridial anaerobic bacteria (NAB) and included PCR of midstream morning urine. Among FAB in the urine and vagina dominated coagulase-negative staphylococci and NAB. Bacterial patterns of studied habitats turned out to be similar in many respects. In the urine Megasphaera spp., Veillonella spp., Prevotella spp., Mobiluncus spp., Fusobacterium spp. were found, whereas in the vagina these microorganisms were not present. Cluster analysis revealed no significant differences in the concentration of the same microorganisms isolated from the urine and vagina. When comparing the frequency of microorganism detection in urine by bacteriological method and by PCR, bacterial patterns were identical in 56% of cases.