Emergence of CTX-M group 1 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strains in the community.

AIM: Molecular characterization of ESBL-producing K. pneumoniae strains isolated from urine of outpatients in Zagreb region during the last five years. METHODS: During the five-year study period a total of 2, 651 K. pneumoniae strains were isolated from urine of nonhospitalized patients with significant bacteriuria. ESBL production was detected by double-disk diffusion technique and by > or = 3-dilution reduction in the minimal inhibitory concentration of ceftazidime in the presence of clavulanate. A total of 441 ESBL-producing K. pneumoniae strains (15.5%) were collected and 17 strains were further characterised. Double-disk synergy test was used to detect ESBLs. Minimum inhibitory concentrations (MICs) were determined by broth microdilution method according to CLSI. The transferability of cefotaxime resistance was tested by conjugation (broth mating method). PCR was used to detect alleles encoding ESBL enzymes. The genotypes of the strains were compared by pulsed-field gel electrophoresis (PFGE) of Xba I-digested genomic DNA. RESULTS: A significant difference in frequencies of ESBL isolates was observed. In the first year of study only 4.9% of isolated strains were ESBL producers, while in the second year 17.% ESBL-positive strains were detected (p < 0.01), and the frequency remained stabile within following years. All strains yielded an amplicon with primers specific for SHV beta-lactamases and CTX-M beta-lactamases. Based on sequencing of bla(CTX-M) genes enzymes of nine strains were identified as CTX-M 15 beta -lactamase and three as CTX-M-14. Isolates were not clonally related. CONCLUSION: The study demonstrated community-associated emergence of CTX-M 1 beta-lactamase-producing K. pneumoniae strains.

[Resistance of uropathogenic strains of Escherichia coli in pregnant women and other women in generative ages in comparison with antibiotics consumption in Zagreb]. / Rezistencija uropatogenih sojeva bakterije Escherichia coli kod trudnica i zena generativne dobi u usporedbi s potrosnjom anti- biotika u Zagrebu.

AIM: To compare resistance of uropathogenic strains of Escherichia coli (UPEC) to antibiotics in women in generative ages and pregnant women during two year period (2004 and 2008) in Zagreb, and comparison of resistance and the consumption of antibiotics. METHODS: The standard disk-diffusion method was used for sensitivity testing to 16 different antibiotics. Data on antibiotic utilization were used to calculate the number of defined daily doses (DDD) and DDD per 1000 inhabitants using Anatomical-Therapeutic-Chemical/DDD methodology. Data on antibiotic consumption during pregnancy were collected using a questionnaire filled in by 893 women after delivery. RESULTS: During 2004 resistance of UPEC to antimicrobial drugs was not different in pregnant and in non-pregnant women, with the exception of amoxicillin and nitrofurantoin, with statistically higher resistance in pregnant women (p < 0.01). Four years later the statistically higher resistance to norfloxacin was observed in non-pregnant women (p < 0.01). Comparing the resistance in 2004 and 2008, in the both groups of women a statistically significant decrease of resistance to cefalexin and nitrofurantoin was detected (p < 0.01). Outpatient utilization of antimicrobial drugs in Zagreb increased significantly, from 32 to 39 DDD/1000 inhabitants per day. The most used antibiotic was co-amoxiclav, and its utilization increased from 9.6 to 12.2 DDD/1000/day. Amoxicillin and co-amoxiclav were used during pregnancy by 9.6% interviewed women. CONCLUSION: The observed significant decrease of resistance to cefalexin makes that antibiotic the drug of choice for treatment of urinary tract infections in women in generative ages, and together with coamoxiclav can be administered in pregnancy. Constant monitoring of urinary tract pathogens resistance to antimicrobial agents ensures the effectiveness of empirical therapy, whose versatile use is limited due the potentially harmful effects of antimicrobial drugs on fetus.

[Frequency of Haemophilus spp. in urinary and and genital tract samples]. / Zastupljenost bakterija roda Haemophilus u uzorcima iz mokracnog i genitalnog sustava.

AIM: To determine the prevalence and antibiotic susceptibility of Haemophilus influenzae and H. parainfluenzae isolated from the urinary and genital tracts. METHODS: Identification of strains bacteria Haemophilus spp. was carried out by using API NH identification system, and antibiotic susceptibility was performed by Kirby-Bauer disk diffusion method. RESULTS: A total number of 50 (0.03%) H. influenzae and 14 (0.01%) H. parainfluenzae (out of 180, 415 samples) were isolated from genitourinary tract. From urine samples of the girls under 15 years of age these bacteria were isolated in 13 (0.88%) and two (0.13%) cases, respectively, and only in one case (0.11%) of the UTI in boys (H. influenzae). In persons of fertile age, it was only H. influenzae bacteria that was found in urine samples of the five women (0.04%) and in three men (0.22%). As a cause of vulvovaginitis, H. influenzae was isolated in four (5.63%), and H. parainfluenzae in two (2.82%) girls. In persons of fertile age, H. influenzae was isolated from 10 (0.49%) smears of the cervix, and in nine (1.74%) male samples. H. parainfluenzae was isolated from seven (1.36%) male samples. (p < 0.01). Susceptibility testing ofH. influenzae and H. parainfluenzae revealed that both pathogens were significantly resistant to cotrimoxasol only (26.0% and 42.9%, respectively). CONCLUSION: In the etiology of genitourinary infections of girls during childhood, genital infections of women in fertile age (especially in pregnant women), and men with cases of epididimytis and/or orchitis, it is important to think about this rare and demanding bacteria in terms of cultivation.

[Characteristics of uropathogens in outpatient catheter-associated urinary tract infections]. / Karakteristike uzrocnika urinarnih infekcija povezanih s kateterima u izvanbolnickoj populaciji.

During the two years period the prevalence of uropathogens responsible for catheter-associated urinary tract infection (CAUTI) in outpatients was investigated including their differences in antimicrobial susceptibility according to the age and gender. Indwelling urinary catheter (IUC) constitutes 0.3% of all processed urine samples. Significant bacteriuria was found in 92.5% of IUC, and polimicrobial ethiology in 63.2%. The most CAUTI (79.3%) was found in elderly male patients (> 65 years). The most frequently isolated uropathogens were Esherichia coli and other Enterobacteriaceae, Pseudomonas aeruginosa, and enterococci. The very high resistance of all Enterobacteriaceae to beta-lactams, fluoroquinolones, and co-trimoxazole was observed with significantly much higher fluoroquinolone (p < 0.01) and co-trimoxazole (p < 0.05) resistance in elderly male patients as compared with younger ones.

Clonal dissemination of highly virulent extended-spectrum beta-lactamase-producing Escherichia coli strains isolated from the urine of non-hospitalised patients in Zagreb region.

Recent data suggest that extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli is an emergent cause of urinary tract infections in non-hospitalised patients in different countries. The aim of this study was to characterise ESBL-producing E. coli strains isolated from the urine of outpatients in the Zagreb region of Croatia. During the 5-month study period, a total of 2451 E. coli strains were isolated from the urine of non-hospitalised patients with significant bacteriuria. A total of 39 ESBL-producing E. coli strains (1.59%) were collected and characterised.

Emergence of uropathogenic extended-spectrum beta lactamases-producing Escherichia coli strains in the community.

The aim of this study was to determine the virulence characteristics and resistance pattern of the extended-spectrum/lactamases (ESBLs)-producing Escherichia coli strains isolated from urine of outpatients in the Zagreb region during a five-month period, and to compare them with the non ESBLs-producing E. coli strains isolated in the same period. Out of 2451 E. coli strains isolated from urine of nonhospitalized patients with significant bacteriuria, a total of 39 ESBLs-producing strains (1.59%) were detected by a double-disk diffusion technique and by the broth-dilution minimal inhibitory concentration reduction method. The 45 non ESBLs-producing strains were randomly chosen, and phenotype of the two groups of strains was characterized and compared. Serogroup O4, hemolysin production, expression of P- and type 1 fimbriae as well as resistance to gentamicin and amikacin were significantly more prevalent characteristics among the ESBLs-producing strains than among non ESBLs-producing strains (p < 0.01), while higher prevalence of trimethoprim-sulfamethoxazole resistance among ESBLs-producing strains was not statistically significant (p > 0.05). Chromosomal DNA analysis by pulsed-field gel electrophoresis exhibited a great genomic similarity among ESBLs-producing strains and revealed that those highly virulent and resistant E. coli strains isolated from urine of outpatients in the Zagreb region had a clonal propagation.

Genital human papillomavirus infection in women from the Zagreb region.

Human papillomavirus (HPV) infection is the most common sexually transmitted infection, especially among young, sexually active individuals. As persistent infection with oncogenic types may lead to cervical cancer, HPV testing is a useful tool to screen for women at risk for subsequent development of cervical cancer. The aim of the study was to determine the prevalence of high-risk HPV (hrHPV) infection in different age groups of cytologically selected women from the Zagreb region, and to evaluate the frequency and results of repeat hrHPV testing. During a one-year study period (November 2005 to November 2006), a total of 3,440 cervical samples from women attending gynecological services of public and private health care systems were received. They were tested for 13 hrHPV genotypes by the polymerase chain reaction based AMPLICOR HPV test (Roche Molecular Systems). The overall prevalence of hrHPV was 34.6%. Most samples were obtained from women aged 21-30 years (44.2%), followed by the 31-40 (27.6%), 41-50 (15.7%), 51-60 (5.3%) and 261 (2.4%) age groups. Out of 3,227 cervical samples obtained from women of known age, 4.9% were obtained from the group of girls younger than 21, in which the highest prevalence of hrHPV (49.4%) was found. A similar prevalence was observed in women aged 21-30 (45.1%). The prevalence gradually decreased with age. During the study period, repeat hrHPV testing was performed in samples from 66 women at different intervals. Out of 28 women that were hrHPV negative on initial testing, only five women turned positive on repeat testing. Out of 38 women that were positive on initial testing, in one-third hrHPV could not be detected on repeat testing. As expected, hrHPV infection was highly prevalent in female adolescents and young women. Further investigation on repeat hrHPV testing is needed to assess virus clearance and rate of newly acquired infection.

Fungal infections of urogenital system.

We analyzed the frequency of isolation of individual fungal species in the samples of urine, vaginal and cervical swabs from 107 patients (72% women and 28% men) aged 16-82 years, who were treated in primary care for cystitis, vulvovaginitis, and cervicitis. The samples were analyzed at the Microbiological Laboratory of Zagreb Institute of Public Health, Croatia, between September 1, 2002 and June 31, 2003. Eight species of yeast were isolated from the samples. Candida (C.) albicans, C. glabrata, and C. krusei were the most common isolates, with the frequency of 61.7%, 10.3% and 8.4%, respectively. Other species (C. guilliermondii, C. famata, C. tropicalis, C. parapsilosis, and C. kefyr) were less frequently isolated, between 0.9% and 7.5%. In women, the frequency of isolation of C. albicans species from urine samples ranged from 83.3% to 30.8%, and from uterine cervical swabs from 85.7% to 50%, showing a decreasing trend with patients' age. The frequency of C. albicans isolates from vaginal swabs was equal in younger (<30 years) and older women (l51 years), but twice less frequent in middle-aged women. In men, the frequency of C. albicans species isolated from urine samples decreased with age from 100% to 52.4%. In the 107 analyzed samples with positive fungal culture, yeast isolations were significantly more frequent than pure culture (93.5%) and had a larger number of colony counts (57%). This suggested that these yeast species might have a pathogenic role in the causation of urogenital system infections.