[Urethral microbiome in patients with pulmonary tuberculosis].

INTRODUCTION: Treatment of tuberculosis involves long-term intake of several antimicrobial drugs, including that with a wide spectrum of action, which may affect the microflora of the urinary tract. AIM: To determine the effect of long-term combination of antibiotic drugs on the microbiome of male urethra. MATERIALS AND METHODS: A total of 75 men were included in an open, prospective, non-comparative study, including 63 patients with pulmonary tuberculosis who received anti-tuberculosis therapy for at least 3 and no more than 5 months without urinary tract infections (main group) and 12 patients with non-infectious urological diseases (urolithiasis, benign prostatic hyperplasia) and normal urinalysis (comparison group). All patients underwent urethral swab with a sterile cotton after cleaning of genital area. The identification of pathogens and the quantitative assessment of composition of the microbiota in the obtained samples was performed by the method of real-time polymerase chain reaction (RT-PCR). The quantitative results are presented in the number of genomic equivalents in 1 ml (GE/ml), which are proportional to the microbial contamination of the samples. RESULTS: "Classical" pathogenic microflora in the urethral swab was detected in 1/3-1/4 patient with tuberculosis. Enterobacteriaceae spp./Enterococcus spp. in a titer of 103-104 GE/ml was identified in 16 (25.4%) patients. Staphylococcus spp. was found in the titer of 103-106 GE/ml in 20 males (31.7%). In the comparison group, Staphylococcus spp. was present in half of the cases (n=6), and Enterobacteriaceae spp./ Enterococcus spp. were isolated in every third patient (n=4) with a titer of 103-105 GE/ml. In patients with tuberculosis, Corynebacterium spp. was most commonly detected (n=31, 49.2%), while in the comparison group this pathogen was isolated only in 3 (25.0%) patients. At the same time, the detection rate of Candida spp. in urethral swab was not significantly different between two groups (7.9 and 8.3%, respectively). CONCLUSION: Patients with tuberculosis receiving combination of chemotherapeutic drugs for at least 3 months, and male without signs of urinary tract infections have significant differences in the spectrum of the urethral microflora.

[Association between early peripheral lung cancer and nontuberculous mycobacterial lung disease]. / Assotsiatsiya rannego perifericheskogo raka i netuberkuleznoi mikobakterial'noi bolezni legkikh.

Mycobacteriosis is an infectious disease caused by nontuberculous mycobacteria, with granulomatous inflammation in the affected organs and tissues. The association of mycobacteriosis and lung cancer in one patient is quite rarely observed and is of interest to the medical community. A 63-year-old male patient was admitted to the hospital with a diagnosis of infiltrative tuberculosis in SI-SII of both lungs in the lung cavitation phase on the right, peripheral mass in SIII on the right in order to make a decision about a surgical intervention. Morphological examination of the material obtained during upper lobectomy revealed mucinous adenocarcinoma concurrent with mycobacterium-associated inflammation with cavern formation. A bacteriological study confirmed the presence of the pathogen Mycobacterium avium. This study reports a case of association of the diseases with successful surgical intervention without complications in the early postoperative period.

[New opportunities of using gialuronidase in chronic prostatitis].

INTRODUCTION: The use of hyaluronidase contributes to improvement in microcirculation, destruction of biofilms and reducing of congestion in patients with chronic prostatitis. Based on this mechanism, the use Longidaza for the diagnosis of latent bacterial prostatitis was suggested. AIM: To evaluate diagnostic and treatment capabilities of the long-acting enzyme preparation Longidaza (rectal suppositories with an activity of 3000 IU), prescribed to patients with chronic prostatitis in routine clinical practice. MATERIALS AND METHODS: A total of 39 men with chronic prostatitis of categories II and IIIa were included in open, prospective, non-interventional, comparative, single-center study. A microbiological study of expressed prostate secretion (EPS) was performed three times, at baseline, after using two suppositories Longidaza 3000 IU and after 7 weeks of therapy. From the 1st to the 10th suppository, Longidaza was used every 48 hours, then, from the 11th to the 20th suppository, it was prescribed every 72 hours. The isolation of the pathogen was an indication for the antibiotic therapy. The treatment outcome was evaluated by using NIH-CPSI scale and laboratory and microbiological analysis of EPS. RESULTS: Diagnostic stage. Initially, pathogens were isolated in 27 (69.2%) patients. The number of leukocytes in EPS averaged 25.9+/-2.3. The use of two suppositories Longidaza increased the number of patients with isolated pathogens to 33 (82%). In 7 out 12 patients with an initial diagnosis of abacterial prostatitis, the use of two suppositories of Longidaza contributed to isolation of the pathogen in EPS (58.3%). The number of leukocytes in the EPS after enzymatic provocation increased by 50.2%. Treatment stage. The number of patients with isolated pathogens decreased from 33 to 7 (17.9%). The pain intensity decreased on average to 2.2+/-0.4 points, and in 27 (69.2%) men dysuria resolved. The quality of life significantly improved up to 1.3+/-0.2 points. The total score on NIH-CPSI scale decreased on average from 17.3+/-1.9 to 3.9+/-0.3 (p<0.05). An increased number of white blood cells in EPS was seen only in 5 (12.8%) men. CONCLUSION: In many cases, bacterial prostatitis remains undiagnosed. Rectal suppositories Longidaza can be recommended for the diagnosis of the latent bacterial chronic prostatitis, since this drug contributes to detection of pathogen and inflammation.

Detection of Mutations in Mycobacterium tuberculosis pncA Gene by Modified High-Resolution Melting Curve Analysis of PCR Products.

We developed a protocol for detection of mutations in the pncA gene associated with M. tuberculosis resistance to pyrazinamide by analyzing melting curves of 7 overlapping amplicons with artificial heteroduplex formation (H-HRM) formed by co-amplification of wild-type DNA and test DNA and compared its efficiency and robustness with those of classical HRM analysis. Using HRM and H-HRM, we analyzed 35 PZAR DNA isolates carrying mutations in the pncA gene, 3 PZAR isolates without mutations in the pncA gene, and 20 PZAS isolates without mutations in the pncA gene were analyzed. The sensitivity and specificity of HRM for detection of mutations in the pncA gene were moderate: 88.57% (CI 73.26%-96.80%) and 82.61% (CI 61.22%-95.05%), respectively. The sensitivity of the H-HRM test was 97.14% (CI 85.08%-99.93%) and specificity was 95.65% (CI 78.05%-99.89%), with a significant improvement in accuracy - 96.55% vs. 93.85% for HRM. In general, despite addition stage of equalizing the concentrations of the test and control mycobacterial DNA, H-HRM showed greater stability and reproducibility at standard settings of the melting curve analysis software.

[Characterization of microbial flora in patients suspected of having urogenital tuberculosis].

INTRODUCTION: In the structure of tuberculosis morbidity, urogenital tuberculosis (UGT) ranks second or third only to pulmonary tuberculosis. Every fourth patient has the stage of cavern formation at first presentation. Seventy five percent of nephrotuberculosis cases co-occur with nonspecific pyelonephritis, which excludes "sterile pyuria". We aimed to determine the microbial spectrum of urine and gonadal appendages in patients with suspected UGT and to investigate the susceptibility of the isolated microorganisms to the main antibacterial drugs. MATERIAL AND METHODS: The study investigated the spectrum of pathogenic microflora isolated from patients admitted to the urogenital clinic of the TB Research Institute from January 1, 2012 to September 30, 2016 to rule out UGT. Microorganism species were identified by time-of-flight mass spectrometry using the microbiological analyzer Phoenix 100 (Becton Dickinson, USA). Antibiotic resistance was determined using the Phoenix 100 microbiological analyzer (Becton Dickinson, USA). RESULTS: A total of 3004 bacteriological tests for nonspecific microflora were performed during the analyzed period. In the structure of pathogens, E. coli comprised about half of the cases thus confirming its role as the leading uropathogen. Only piperacillin/tazobactam and nitrofurantoin provided relatively plausible antimicrobial resistance rates, for other antibacterial drugs the rates of susceptible strains progressively decreased and by 2016 dropped to 40% and lower. CONCLUSION: Our findings cannot be extrapolated to the entire population since patients presenting for differential diagnosis receive multiple courses of treatment with various antibacterial drugs, which led to the selection of resistant strains. The findings emphasize the need for a strict selection of antibiotics for urogenital infection empirical therapy. In the absence of response, the patient should be immediately examined for tuberculosis using molecular and genetic diagnostic techniques.

Detection and Characteristics of Rifampicin-Resistant Isolates of Mycobacterium tuberculosis.

Genotyping and analysis the drug resistance of 59 isolates of M. tuberculosis obtained from patients living in Altai Territory were performed using a BACTEC MGIT 960 fluorometric system by means of VNTR typing (variable number tandem repeat), PCR-RFLP analysis, and sequence analysis. The occurrence frequency was highest for isolates of the Beijing family (n=30, 50.8%). Analysis of mutation spectrum in the rpoB gene associated with rifampicin resistance revealed the major mutation (codon 531 of the rpoB gene) in 93% samples, which allows us to use rapid test systems.

Complete Genome Sequence of a Novel Clinical Isolate, Mycobacterium abscessus Strain NOV0213.

Mycobacterium abscessus is a rapid-growing species of nontuberculous mycobacteria that is frequently associated with opportunistic infections in humans. We determined the complete genome sequence of the M. abscessus strain NOV0213, which was isolated from a patient with tuberculosis-like disease and with various antibiotic resistances.

[Diagnosis and treatment of cystitis: more questions than answers?]

INTRODUCTION: Despite the prevalence of acute cystitis, there are still many unsolved problems of diagnosis and treatment of this disease. MATERIAL AND METHODS: To determine the nosological structure of dysuria, 126 female patients who sought medical attention for frequent painful urination were examined. To determine the incidence of outpatient visits to an urologist for patients with cystitis, medical records of 6753 patients of municipal outpatient clinic were analyzed. The results of treating cystitis in 85 patients also were evaluated. RESULTS: Among 126 patients with dysuria, 31 (24.6%), 42 (33.3%) and 47 (37.3%) patients had acute uncomplicated cystitis, recurrent (chronic) cystitis without the complicating factors and recurrent (chronic) cystitis with complicating factors, respectively. Three (2.4%), 2 (1.6%) and 1 (0.8%) patients had trichomoniasis, urogenital herpes and tuberculosis, respectively. As a result of 6753 visits to the urologist, inflammatory diseases of the genitourinary system were detected in 3194 (47.3%) patients, of which 64.7%, 19.6%, 5.3% and 2.7% had chronic pyelonephritis, chronic cystitis, chronic prostatitis and acute cystitis, respectively. Selecting the optimal drugs increases the treatment effectiveness of acute uncomplicated and chronic cystitis by 6% and 17%, respectively. CONCLUSIONS: Most of clinical guidelines and scientific publications aimed at acute uncomplicated cystitis, while the proportion of the disease does not exceed 26% among the patients presenting with dysuria, and comprise only 2.7% among outpatient urologist visits for infectious and inflammatory diseases of the urogenital tract. Selecting the optimal drugs increases the treatment effectiveness of acute uncomplicated and chronic cystitis by 6% and 17%, respectively.

[INCIDENCE OF NOSOCOMIAL UROPATHOGENS AND DYNAMICS OF THEIR SENSITIVITY].

In order to determine the basic spectrum of urinary tract infections and their sensitivity to modern antimicrobials, the 5-year (2009-2013) study of 2621 isolates ofuropathogens from inpatients of two centers of Novosibirsk and Barnaul was carried out. Instability of proportion of the isolated microorganisms both in time and in two compared neighboring regions was found. The role of E. coli was insignificant: in Barnaul the maximum share E. coli among all uropathogens was 56.1% in 2009 (with a five-fold drop in the next year to 10.5%), Novosibirsk share of E. coli increased from 28,1% in 2009 to 49.4% in 2013, with an unaccountable fall to 33.5% in 2011. In evaluating the resistance of E. coli, many groups have shown a steady growth in antibiotic resistance. Resistance of microflora to 2nd generation cephalosporins increased by 3 times, to ciprofloxacin - by 2 times and to amoxicillin - by 7 times. However, there were virtually no strains resistant to carbapenems. Zero resistance to fosfomycin was found - all strains were susceptible to this antibiotic within 5 years. Range of UTI pathogen species in two adjacent regions was subject to significant fluctuations, no patterns were identified. In contrast, the level of microflora resistance increased. Within 5 years, it increased to basic antibiotics by 7 times, while maintaining a low resistance to nitrofurantoin, gentamicin and imipenem, and zero resistance to fosfomycin. The results of this study are consistent with international data on the partial loss of E. coli role as the leading uropathogen. Gram-positive cocci are becoming increasingly important, which is worth considering when choosing a drug for empiric therapy. Optimal antibiotics and uroseptics for treatment of patients with acute uncomplicated cystitis remain fosfomycin and nitrofurantoin, as was shown earlier.

[DIAGNOSIS OF UROGENITAL TUBERCULOSIS].

The sequential algorithm of diagnosis of tuberculosis of the urogenital system with emphasis on the identification of the causative agent is presented; the approach to the examination of the patient is described in detail; regimens of ex juvantibus treatment are provided.

[Revisiting the causes of low detection of M. tuberculosis in urine].

The evaluation of potential anti-TB activity and bactericidal activity against opportunistic enterobacteria in urine of healthy people using the automated BACTEC MGIT 960 system for cultivation and determination of drug resistance of mycobacteria and culture on solid media was performed. It has been established that the urine of healthy people do not have bactericidal activity against M. tuberculosis and E. coli in vitro. The one of the possible reasons for the low detection of the pathogen in urogenital tuberculosis--asymptomatic bacteriuria--was identified.

Highest prevalence of the Mycobacterium tuberculosis Beijing genotype isolates in patients newly diagnosed with tuberculosis in the Novosibirsk oblast, Russian Federation.

In order to assess the genetic diversity of Mycobacterium tuberculosis in the Novosibirsk Region and determine profiles of resistance, 106 M. tuberculosis isolates were analysed. Fifty (47 %) isolates were identified using variable number tandem repeat typing as being in the Beijing family, of which eight (16 %) were type M2 isolates with the genetic profile 233325153533424 and eight (16 %) were type M11 isolates with the genetic profile 233325173533424, both of which are widespread in Russia. Mutations associated with resistance to isoniazid and rifampicin were identified. Of 48 isolates with resistance to isoniazid, 42 (87.5 %) contained a Ser(315)→Thr substitution in the katG gene and one contained a T→A substitution at position -34 of the promoter region of the ahpC gene. Of 31 isolates with resistance to rifampicin, 19 (61 %) each contained a mutation (TCG→TTG) at codon 531 of the rpoB gene. Two isolates each contained a mutation (GAC→GTC) at codon 516 of the rpoB gene and two others each contained a substitution at codon 526 of the rpoB gene, leading to a His→Asn substitution in one case and a His→Asp substitution in another case. One isolate contained a mutation (CTG→CCG) at codon 533 of the rpoB gene. An association between the Beijing genotype and multidrug resistance was demonstrated (R = 0.2, P = 0.032). However, it was interesting to note that a significant proportion (46 %) of isolates were sensitive to all drugs tested.