A twelve-year retrospective analysis of prevalence and antimicrobial susceptibility patterns of Ureaplasma spp. and Mycoplasma hominis in the province of Lower Silesia in Poland.

OBJECTIVE: Genital mycoplasmas are opportunistic pathogens that have been associated with urogenital infections in humans. Only a few groups of antimicrobials are available for treatment of urogenital tract infections caused by genital mycoplasmas. However, emerging resistance of mycoplasmas to antimicrobial agents has been reported worldwide. The aim of the study was a retrospective analysis of the prevalence and antimicrobial susceptibility patterns of M. hominis and Ureaplasma spp. in patients with urogenital tract infections during a twelve-year period between 2003 and 2015. STUDY DESIGN: Mycoplasma IST2 test was used for the detection, enumeration, identification and antimicrobial susceptibility testing of genital mycoplasmas in 1182 samples from 778 women and 404 men with genitourinary tract infection. Indicative enumeration in the test determines whether the mycoplasma count in the sample is equal or higher than the threshold set at 104 colony forming units. RESULTS: A total of 152 (12.8%) samples were found to be positive for genital mycoplasmas. M. hominis was detected only in three samples and Ureaplasma spp. in 141 samples. Both, M. hominis and Ureaplasma spp. were detected in the remaining eight samples. In the analyzed period between 2003 and 2015, a gradually increasing resistance of ureaplasmas to ciprofloxacin and clarithromycin and decreasing resistance to ofloxacin, erythromycin and tetracycline were observed. Pristinamycin, josamycin and doxycycline were most active against Ureaplasma spp. In contrast, fluoroquinolones had the lowest efficacy against Ureaplasma spp. and as many as 116 (82.3%) and 77 (54.6%) of Ureaplasma spp. isolates were resistant to ciprofloxacin and ofloxacin, respectively. M. hominis isolates were uniformly resistant to azithromycin, clarithromycin and erythromycin but susceptible to josamycin, ofloxacin, doxycycline and pristinamycin. One-third of these isolates were resistant to ciprofloxacin and tetracycline. CONCLUSION: In the study Ureaplasma spp. and M. hominis were detected with relatively low frequency in comparison with other studies however, most of these isolates were resistant to ciprofloxacin indicating the need for better management of ciprofloxacin prescription. Important limitations of Mycoplasma IST2 assay concerning antimicrobial susceptibility testing and divergences between breakpoints in the test and EUCAST guidelines point the need to introduce new methodologies to improve evaluation of resistant strains at our region.

Mycoplasma pulmonis of Rodents as a Possible Human Pathogen.

Mycoplasma pulmonis is a naturally occurring respiratory pathogen in rodents. To date, this pathogen was not isolated from humans. This study aimed to evaluate the prevalence and seropositivity to M. pulmonis in humans who have had direct contact with rats. Moreover, the prevalence of M. pulmonis in pet and laboratory rats was assessed. Overall, 131 and 235 oropharyngeal swab samples were collected from human individuals and rats, respectively. In humans, M. pulmonis was detected by PCR in 21 of 86 pet rat keepers (24.42%), 10 of 13 technicians (76.32%), and 8 of 32 (25.0%) veterinarians. In rats, M. pulmonis was identified by PCR in 86 of 122 pet rats (70.49%) and 56 of 113 (49.56%) laboratory rats. Seroprevalence in humans was examined by screening sera from 44 individuals for M. pulmonis-specific IgG using ELISA. In total, 26 out of 44 (59.09%) humans were seropositive to M. pulmonis (4 out of 9 technicians, 8 of 12 veterinarians, and 15 of 23 pet rat keepers).The high antibody titer was found in 4 individuals (2 pet rat keepers and 2 veterinarians), whereas the moderate and low antibody titers were found in 8 and 14 individuals, respectively. The high antibody titer found in humans might indicate an active infection. However, it is unknown whether the presence of M. pulmonis in humans might be associated with disease and whether the foreign Mycoplasma can survive for long in its new environment.

Relevance of serology for Mycoplasma pneumoniae infection among children with persistent cough.

BACKGROUND: Mycoplasma pneumoniae is an important cause of upper and lower respiratory tract infections. Cough and tracheobronchitis are the commonest features of M. pneumoniae infection but diagnosis based on clinical symptoms that may be due to other respiratory pathogens is impossible. Thus laboratory testing for M. pneumoniae is particularly important. Correct and rapid diagnosis of M. pneumoniae infections is of prime importance to introduce appropriate antibiotic treatment. OBJECTIVES: Evaluation of the incidence of IgM and IgG antibodies specific to M. pneumoniae among children with pneumonia and/or chronic cough. MATERIAL AND METHODS: Serum samples from 148 children with a history of chronic cough (lasting at least one month), recurrent respiratory tract infections, allergic rhinitis, and/or inflammatory changes on X-chest ray. First, all sera were screened for specific anti-M. pneumoniae antibodies using agglutination test following the detection of specific IgM and IgG anti-M. pneumoniae antibodies using immunoenzymatic assays. RESULTS: Out of the 148 serum samples, 57 (38.5%) gave positive screening results. However, the presence of M. pneumoniae-specific IgM and/or IgG antibodies was confirmed by immunoenzymatic assays in only 30 (52.6%) of these 57 positive samples. These results indicated that in as many as 27 (47.4%) out of the 57 serum samples screened, false-positive results occurred. CONCLUSIONS: Evaluation of acute- and convalescent-phase sera is necessary to make possible accurate interpretation of the serological testing results.

Colonization of the lower urogenital tract with Ureaplasma parvum can cause asymptomatic infection of the upper reproductive system in women: a preliminary study.

PURPOSE: Genital ureaplasmas are considered opportunistic pathogens of human genitourinary tract involved in adverse pregnancy sequelae and infertility. While association of Ureaplasma urealyticum with urogenital tract infections is well established, the role of Ureaplasma parvum in these infections is still insufficient. In the study, we compared how often cervicovaginal colonization with U. parvum is associated with the presence of these microorganisms in the upper genitourinary tract of fertile and infertile women. METHODS: We used PCR assay to determine the prevalence of U. parvum and U. urealyticum in pairs of specimens, i.e., vaginal swabs and Douglas' pouch fluid samples from consecutive 40 women with no symptoms of genital tract infection. RESULTS: In total, 19 (47.5 %) of the 40 samples were positive for ureaplasmas. U. parvum was simultaneously detected in pairs of samples in five (55.5 %) of the nine (47.4 %) women positive in PCR assay. As many as 5 (18.5 %) of the 27 infertile women and 1 (7.7 %) of the 13 fertile women showed infection of the upper genital tract with U. parvum. CONCLUSION: The results of the study demonstrated that colonization of the lower genital tract with U. parvum can produce asymptomatic infection of the upper reproductive system in women. These findings also imply that U. parvum may be present in the upper genital tract at the time of conception and might be involved in adverse pregnancy outcomes.

ESBL-producing Escherichia coli isolated from children with acute diarrhea - antimicrobial susceptibility, adherence patterns and phylogenetic background.

BACKGROUND: Escherichia coli remains the principal bacterial pathogen in childhood diarrhea and constitutes an important public health problem, especially in developing countries. Diarrheagenic E. coli strains often display resistance to beta-lactams due to the production of extended-spectrum beta-lactamases (ESBLs). OBJECTIVES: A total of thirty ESBL-producing E. coli strains colonizing the gastrointestinal tracts of children with acute diarrhea were studied in order to determine their antimicrobial susceptibility, adherence patterns to the HEp-2 cell line and phylogenetic background. MATERIAL AND METHODS: ESBL production was detected by the double disk synergy test (DDST). The minimal inhibitory concentrations (MICs) of antibacterial drugs were determined by an agar dilution technique on Mueller-Hinton agar. The presence of bla(TEM), bla(SHV) and bla(CTX-M) determinants in the strains studied was ascertained by polymerase chain reaction (PCR). RESULTS: The strains displayed the resistance pattern typical of ESBL producers. The majority of them (23 out of 30) were found to produce CTX-M-type ESBLs conferring a high level of resistance to oxyimino-beta-lactams, especially to cefotaxime and ceftriaxone. In many cases, the strains exhibited resistance to non-beta-lactam antimicrobials, such as gentamicin, amikacin, co-trimoxazole and tetracycline. On the other hand, these strains were uniformly susceptible to carbapenems, to oxyimino-beta-lactams combined with clavulanic acid and to tigecycline. The E. coli strains were distributed among the four main phylogenetic groups: A, B1, B2 and D. The in vitro adhesion assay revealed that all but two of the strains adhered to the HEp-2 epithelial cell line. Aggregative and diffuse adherence patterns were found to be the most prevalent. CONCLUSIONS: CTX-M-type enzymes were the most prevalent ESBLs among the strains studied. As many as 40% of the diarrheagenic E. coli isolates were found to belong to phylogenetic group D, which usually comprises E. coli strains associated with extra intestinal infections. The effectiveness of tigecycline against ESBL-producing E. coli strains was similar to that of imipenem and meropenem.

Virulence genes profiles and phylogenetic origin of Escherichia coli from acute and chronic intestinal diseases revealed by comparative genomic hybridization microarray.

The association between Escherichia coli virulence factors and chronic intestinal disorders is mostly unknown. The presented study compared the distribution of virulence genes and phylogroups among E. coli isolated from chronic intestinal disorders such as Crohn's disease and irritable bowel syndrome (IBS) with strains isolated from patients with acute diarrhea as a control group. The presence of 159 virulence genes corresponding to known E. coli pathotypes was determined among 78 E. coli archive strains isolated from IBS, acute diarrhea and Crohn's disease using CGH microarray. E. coli isolated from IBS demonstrated a mosaic of virulence genes specific to enteropathogenic, enterotoxigenic, enterohemorrhagic E. coli strains and Shigella species. In contrast, virulence factors and phylogroups distribution among E. coli isolated from children with acute diarrhea was similar to extraintestinal E. coli strains that probably acquired some virulence genes. The acquisition of virulence genes might have an impact on diarrheagenic potential of these strains. On the other hand, E. coli isolated from children with Crohn's disease seem to be similar to adherent-invasive E. coli strains (AIEC), as it lack most known virulence genes. The presented study showed that these analyzed groups of E. coli strains differed from each other with the respect to the distribution of virulence genes. The differences in gene content support the idea that the participation of E. coli in chronic intestinal diseases is mostly related to virulence potential of these strains.

Invasive properties, adhesion patterns and phylogroup profiles among Escherichia coli strains isolated from children with inflammatory bowel disease.

BACKGROUND: A great deal of evidence indicates a link between Escherichia coli (E. coli) and Crohn's disease in adult patients, but there is lack of information on the association of these bacilli with inflammatory bowel disease (IBD) among children. OBJECTIVES: The study was carried out to determine the distribution of phylogenetic group, the adherence patterns and invasive properties of E. coli isolated from children with IBD and non-IBD chronic bowel diseases. MATERIAL AND METHODS: A total of 22 E. coli isolated from biopsy specimens from children with IBD and 21 E. coli strains obtained from children with indeterminate colitis and intestinal polyps were examined for adherence and internalization to the Int407 cell line. Genes involved in epithelial cell invasion and genes specific to E. coli phylogroups were determined by polymerase chain reaction (PCR). RESULTS: The undefined adherence pattern predominated among the isolated E. coli, although most of them demonstrated the afaD and aggB genes encoding invasions of diffusely adhering and enteroaggregative E. coli. Regardless of the clinical entity, most E. coli were internalized by Int407 epithelial cells and belonged to the B2 and D phylogroups. CONCLUSIONS: The wide distribution of adhesive E. coli capable of entering Int407 cells but also having genes encoding adhesins and invasins characteristic to pathogenic E. coli strains seems to indicate that these E. coli may represent a large group of pathogenic E. coli strains contributing to chronic intestinal disorders.

Characterization of genes associated with internalization of enteroaggregative Escherichia coli.

On animal models enteroaggregative Escherichia coli (EAEC) can cause mild, but significant mucosal damage, suggesting the invasive capability of these strains. In the study we investigated the ability of typical, aggR-positive and atypical, aggR-negative EAEC isolates to enter intestinal epithelial Int407 cells in relation to the distribution of genes encoding the putative invasins described among pathogenic E. coli categories. The results demonstrated that regardless of origin and affiliation to typical and atypical EAEC, most isolates examined were internalized by the epithelial cells to different extent. Although as many as 50 (84.3%) EAEC demonstrated a variety of combinations of the aggB, afaD, ipaH and tia genes determined, there was no correlation between the invasion efficiency of these strains and the presence of any particular gene involved in invasion. Most of EAEC examined belonged to phylogenetic group B2 and D.

Occurrence of Mycoplasma genitalium in fertile and infertile women.

OBJECTIVE: To determine the frequency of occurrence of Mycoplasma genitalium in the reproductive organs of infertile women in comparison with a control group of healthy, fertile women. DESIGN: Prospective study. SETTING: Gynecology Clinic at the 2nd Department of Gynecology and Obstetrics of the Wroclaw Medical University, Poland. PATIENT(S): The study included 51 patients with primary infertility (24 women with idiopathic infertility) and 23 women with proven fertility. INTERVENTION(S): Cervical smear and smear from the peritoneal cavity, performed during laparoscopy. MAIN OUTCOME MEASURE(S): Presence of the genetic material of M. genitalium in the collected material analyzed using polymerase chain reaction (PCR). RESULT(S): M. genitalium was found in the cervical canal of 19.6% of all infertile patients and in 4.4% of fertile patients. In addition, the pathogen was discovered in the cervical canal of 29% patients with unexplained (idiopathic) infertility, which in comparison with the fertile group was a statistically significant difference. In the abdominal cavity, M. genitalium was found in 5.8% of patients from the infertile group (in 8.4% patients with idiopathic infertility), whereas it was not detected in the material obtained from the studied fertile patients. CONCLUSION(S): The results obtained may suggest that M. genitalium is a species having an impact on impaired fertility in women.

[Frequency of detection of Ureaplasma urealyticum and Mycoplasma hominis in cervical canal and the Douglas pouch of infertile and fertile women]. / Czestosc wykrywania Ureaplasma urealyticum i Mycoplasma hominis w kanale szyjki macicy i jamie otrzewnej u plodnych i nieplodnych kobiet.

The group of organisms commonly referred to as genital mycoplasmas comprise species most often found in genitourinary tract of sexually active adults as common commensal inhabitants, or pathogens which can possibly cause many different pathologies like: non-gonococcal urethritis, bacterial vaginosis, cervicitis, endometritis or pelvic inflammatory disease. The problem of their morbidity and the possible influence they have on human fertility is still not clear. The aim of this study was to find out whether two investigated species- Ureaplasma urealyticum and Mycoplasma hominis can be detect more often in a group of infertile women. 74 women participated in the study and were assigned to one of 2 groups of patients: infertile women and fertile women without any sign of genital tract infection. Swabs from the cervical canal of the uterus and the fluid from the Douglas pouch were taken during the gynecological examination and laparoscopic procedure. Two diagnostic methods were used: biochemical method- commercial diagnostic kit- Mycoplasma IST 2 and PCR method. The results showed that Ureaplasma urealyticum and Mycoplasma hominis were detected among both fertile and infertile women with nearly the same frequency, much more often in cervical canal than in the Douglas pouch. Ureaplasma urealyticum was more common pathogen than Mycoplasma hominis in both groups and locations. The achieved results point out that the role of genital mycoplasmas in human infertility is still unclear and require further investigations.