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.

[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.

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.