Long-term stability of the urogenital microbiota of asymptomatic European women.

BACKGROUND: To date, information on healthy female urinary microbiota is available mostly at genus level and at one time point. However, profound species-level characterization of healthy urinary microbiome and its stability over time are essential for further correct interpretation of its role in healthy urogenital tract. In this study, we investigated female urogenital microbiome (FUM) at two timepoints (within 2.5-year interval) in young asymptomatic European women. We used culturomics with accurate isolates' identification (MALDI-TOF MS and gene markers sequencing) to understand species stability within healthy FUM. RESULTS: Extended culturomics of voided midstream urine sample pairs revealed a mean Shannon diversity index of 1.25 and mean of 19 species/sample (range 5-39 species; total of 115 species; 1830 isolates). High overall species variability between individuals was captured by beta diversity and a variety of community structure types, with the largest cluster characterized by Lactobacillus crispatus, often in combination with Gardnerella vaginalis or Gardnerella genomospecies 3. Significant FUM composition differences, related to Finegoldia magna and Streptococcus anginosus, according to smoking status were found. A high species variability within individuals (Shannon index SD > 0.5 in 7 out of 10 sample pairs) with a mean of 29% of shared species (range 9.1-41.7%) was observed. Moreover, 4 out of 10 sample pairs clustered in the same community structure type. The stable FUM sample pairs presented high abundance of Lactobacillus crispatus, Streptococcus agalactiae or Lactobacillus paragasseri and Bifidobacterium spp.. Moreover, Gardnerella vaginalis, Gardnerella genomospecies 3 or Gardnerella swidsinskii were often maintained within individuals in high abundance. CONCLUSIONS: Shift in species composition at two distant timepoints was frequently observed among urogenital microbiome of European asymptomatic women. This suggests possible interchange of particular species in healthy FUM and the existence of multiple health-associated FUM compositions in certain individuals. Additionally, we provided additional evidence on resilience of particular bacterial communities and identified certain species more prone to persist in urogenital tract. This study revealed important details on the FUM composition complexity relevant for studies aiming to understand microbiota role in the urogenital tract health and for identification of eubiotic and dysbiotic FUM.

Application of a Pneumococcal Serotype-specific Urinary Antigen Detection Test for Identification of Pediatric Pneumonia in Burkina Faso.

BACKGROUND: Serotype-specific diagnosis of pneumococcal community-acquired pneumonia in children under age 5 years would mark a major advancement for understanding pneumococcal epidemiology and supporting vaccine decision-making. METHODS: A Luminex technology-based multiplex urinary antigen detection (UAD) diagnostic assay was developed and subsequently validated in adults, but its applicability to children is unknown. This study aimed to set appropriate cutoffs for use of the UAD in a healthy pediatric population and apply these cutoffs in children with pneumonia in sub-Saharan Africa. The cutoffs were determined by assessing 379 urines obtained from healthy children under age 5 years from the Bobo-Dioulasso area for serotypes included in 13-valent pneumococcal conjugate vaccine (UAD-1) and the 11 other serotypes unique to 23-valent pneumococcal polysaccharide vaccine (UAD-2). RESULTS: Based on the assigned cutoff values, among 108 children who met the World Health Organization consolidation endpoint criteria, UAD-1 and UAD-2 were positive in 23.3% and 8.3%, respectively; among 364 children with clinically suspected pneumonia who did not meet the World Health Organization criteria, UAD-1 and UAD-2 were positive for 6.6% and 3.6%, respectively. Pneumococcal carriage prevalence was similar among pneumonia cases (30%) versus controls (35%) as was semiquantitative carriage density. CONCLUSIONS: UAD-1 and UAD-2 were able to distinguish community controls from children with pneumonia, particularly pneumonia with consolidation. Future studies are needed to confirm these results and more fully assess the contribution of pneumococcal carriage and concurrent viral infection.

Active surveillance in response to the identification of a single carbapenemase-producing Escherichia coli at a Japanese university hospital.

This report described the experience of active surveillance culture implemented in response to the identification of a single carbapenemase-producing Escherichia coli in a Japanese university hospital. It revealed a horizontal transmission event and an additional asymptomatic carrier of carbapenemase-producing Escherichia coli with unique drug susceptibility and resistance gene profiles. Early implementation of active surveillance culture as a part of multifaceted infection control measures appeared to be useful to control further transmission of carbapenemase-producing Escherichia coli even in the low endemic facility. Further investigations on the timing and usefulness of active surveillance culture in the control of carbapenemase-producing Enterobacteriaceae would be warranted.

Prevalence of urinary colonization by extended spectrum-beta-lactamase Enterobacteriaceae among catheterised inpatients in Italian long term care facilities.

BACKGROUND: Long Term Care Facilities (LTCFs) play a key role in guaranteeing care to patients in developed countries. Many patients, mostly elderly, access LTCFs at some time in their lives, and their healthcare pathways often require them to move back and forth between hospital and outpatient settings. These patterns bring about new challenges regarding infection control, especially healthcare associated infections. METHODS: A point prevalence study was conducted in 23 Italian LTCFs, to identify colonization in patients with urinary catheter (>24 hours). Species identification, susceptibility tests and extended spectrum beta lactamase (ESBL) production screenings were performed using Vitek 2 System. Enterobacteria identified by Vitek 2 System as ESBL-producers or suspected AmpC hyperproducers on the basis of cephamycin resistance, were sent to a research laboratory where they underwent a double-disk synergy test. Finally, ESBL-producers were screened for bla resistance genes by PCR assay. RESULTS: 211 patients with catheter were screened, 185 out of 211 patients showed positive samples for the presence of Enterobacteriaceae, 114 of these 185 patients were colonized by extended spectrum cephalosporins resistant microorganisms. We identified a total of 257 Gram negative pathogens, of which 51.8% (133/257) were extended spectrum cephalosporins resistant. 7 out of 133 cephamycin not susceptible strains proved to be AmpC-type beta-lactamases and 125/133 ESBL-producers; 1 was not further characterized. 43 out of 257 (16.7%) E. coli, 37/257 (14.4%) P. mirabilis, 20/257 (7.8%), P. stuartii, 14/257 (5.4%) M. morganii, 7/257 (2.7%), K. pneumoniae, 4/257 (1.6%) C. koseri proved to be overall ESBL-producers by double-disk synergy test. Third and fourth generation cephalosporin resistant P. mirabilis, P. stuartii and M. morganii strains mainly harboured a blaTEM gene (95.9%), while 89.1% of E. coli were positive for the blaCTX-M determinant by PCR and sequencing. Patients with decubitus had a higher risk of colonization by at least one resistant isolate (p < 0.01). Samples of patients undergoing antibiotic therapy and patients with decubitus showed a higher risk (p < 0.05) of colonization by beta-lactam resistant microorganisms. CONCLUSIONS: These data confirm the presence of high percentages of ESBL-positive Enterobacteria in Italian LTCFs and the predominance of CTX-M type ESBL in E. coli. The alarming presence of ESBL-producing Enterobacteriaceae in Italian LTCFs can seriously compromise the effectiveness of antibiotic therapy.acilities (LTCFs), Antimicrobial resistance.

[Extra-digestive reservoir of extended-spectrum beta-lactamase producing Enterobacteriaceae in non-infected patients: a study based on the systematic searching in urine and other clinical samples]. / Réservoir extra-digestif d'entérobactéries productrices de bêtalactamase à spectre élargi chez des patients non infectés : étude à partir d'une recherche systématique dans les urines et d'autres prélèvements à visée diagnostique.

Our objective was to assess the extra-digestive reservoir of extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBLE) by identifying them in clinical samples in which growing bacteria are not systematically analysed (bacterial identification and antimicrobial susceptibility testing) in routine practice. During a 5-week period, an analysis was systematically performed for Enterobacteriaceae colonies isolated in certain clinical samples (urine samples, respiratory-tract samples, and a group of samples called "miscellaneous samples") according to our study protocol. Samples in which an antimicrobial susceptibility testing was performed for bacteria isolated according to our routine practice were considered infected. Samples in which ESBLE were sought and isolated according to the study protocol were considered colonised. During the study, 2312 urine samples, 327 respiratory-tract samples, and 1887 miscellaneous samples were addressed to the laboratory. Among the 114 urine samples colonised with Enterobacteriaceae, 13 (11.4%) grew with ESBLE whereas this proportion was 5.1% (35/682) in infected US (P<0.01). Among respiratory-tract and miscellaneous samples, 3 ESBLE were isolated in the 55 samples colonised with at least one Enterobacteriaceae. Overall, the systematic searching of ESBLE in the clinical samples provided a 27.7% increase of the patients identified as carriers in the entire hospital during the study period. Further studies would be useful to evaluate the interest of using such a strategy instead of systematic rectal screening.

Extended-spectrum ß-lactamase-producing Escherichia coli strains in the feces of carriers contribute substantially to urinary tract infections in these patients.

INTRODUCTION: The current increase in the incidence of urinary tract infections (UTIs) worldwide caused by extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli may be due to the high number of ESBL-producing Enterobacteriaceae carriers in the community. However, whether ESBL-producing bacteria can cause UTIs in carriers remains uncertain. MATERIALS AND METHODS: In this study, 21 fecal carriers of ESBL-producing Enterobacteriaceae were assessed for UTIs caused by ESBL-producing E. coli. Bacterial isolates obtained from patients' urine and stool specimens were phenotypically and genotypically examined. Clonal similarities of isolates were assessed by multilocus sequence typing (MLST) and random amplified polymorphic DNA (RAPD) fingerprinting. RESULTS: The study revealed that 9 of 21 carriers developed UTIs, and genetic analysis showed that 44% of the UTIs developed were caused by the same ESBL-producing E. coli as that found in the feces of the patients. CONCLUSIONS: The ESBL-producing E. coli in carriers can cause UTIs.

A prospective study of urinary pneumococcal antigen detection in healthy Karen mothers with high rates of pneumococcal nasopharyngeal carriage.

BACKGROUND: Detection of Streptococcus pneumoniae C-polysaccharide in urine is a useful rapid diagnostic test for pneumococcal infections in adults. In young children, high rates of false positive results have been documented due to detection of concurrent nasopharyngeal pneumococcal carriage. The relationship between pneumococcal carriage and urinary antigen detection in adults from developing countries with high pneumococcal carriage prevalence has not been well established. METHODS: We nested an evaluation of the BinaxNOW S. pneumoniae test within a longitudinal mother-infant pneumococcal carriage study in Karen refugees on the Thailand-Myanmar border. Paired urine and nasopharyngeal swab specimens were collected from 98 asymptomatic women at a routine study follow-up visit. The urine specimens were analyzed with the BinaxNOW test and the nasopharyngeal swabs were semi-quantitatively cultured to identify pneumococcal colonization. RESULTS: 24/98 (25%) women were colonized by S. pneumoniae but only three (3%) had a positive BinaxNOW urine test. The sensitivity of the BinaxNOW test for detection of pneumococcal colonization was 4.2% (95% CI: 0.1-21.1%) with a specificity of 97.3% (95% CI: 90.6-99.7%). Pneumococcal colonization was not associated with having a positive BinaxNOW test (odds ratio 1.6; 95% CI: 0.0-12.7; p=0.7). CONCLUSIONS: Significant numbers of false positive results are unlikely to be encountered when using the BinaxNOW test to diagnose pneumococcal infection in adults from countries with moderate to high rates of pneumococcal colonization.

MLVA is a valuable tool in epidemiological investigations of Escherichia coli and for disclosing multiple carriage.

BACKGROUND: Escherichia coli is a common cause of healthcare-associated urinary tract infection, and is frequently present in the urine of elderly people. Transmission of E. coli between individuals has been suggested, and individuals can be concurrently colonized with several types. Efficient typing methods are required to investigate these epidemiological relationships, and we have examined the applicability of multiple-locus variable number tandem repeat analysis (MLVA). METHODS: Up to 20 E. coli isolates were sampled per individual from 30 elderly residents at 2 long-term care facilities, and typed using MLVA, pulsed-field gel electrophoresis (PFGE) and PhenePlate (PhP). RESULTS: Thirty-one E. coli types were identified using MLVA, compared to 38 and 32 using PFGE and PhenePlate, respectively. All isolates were typeable using MLVA and PhenePlate, whereas PFGE failed to type isolates from 2 individuals. The Wallace 1 coefficient indicated a high probability that isolates of the same PFGE type were also of the same type according to the other 2 methods. However, the Wallace 2 coefficient indicated a low probability that isolates of the same PhP type would be classified as the same type by PFGE. Twenty-four of the MLVA types were uniquely restricted to single individuals, whilst 7 MLVA types were found in more than 1 individual. Colonization with more than 1 MLVA type was seen in 8 individuals. There was no evidence of specific institutional types at either of the 2 long-term care facilities. CONCLUSION: MLVA displays a high discriminatory power, and shows substantial potential with respect to epidemiological studies and infection control issues.

[Is it possible to detect Staphylococcus aureus colonization or bacteriuria before orthopedic surgery hospitalization?]. / Le dépistage en ambulatoire des patients porteurs de Staphylococcus aureus ou présentant une colonisation urinaire et devant bénéficier d'une chirurgie orthopédique est-il réaliste ?

AIM OF THE STUDY: Evaluate the feasibility of Staphylococcus aureus nasal colonization and bacteriuria screening in outpatients before realizing a decolonization treatment in S. aureus carriers and a bacteriuria treatment before hospitalization. METHODS: All patients undergoing hip, knee or back surgery in which prosthesis were implanted between October 2007 until the end of June 2008 were included. Microbiological studies were performed before hospitalization. Notice for S. aureus decolonization regimen was delivered to each patient and to the general practitioner only if the patient had nasal carriage. RESULTS: Only 91.2% (240/263) of patients had microbiological results. Prevalence of S. aureus colonization was 21.4% (48 positives/224). Three patients were colonized with methicillin-resistant staphylococci. Decolonization regimen was applied before surgery to 70.8% (n=34) of the colonized patients. Among the patients, 8.9% (20/225) had bacteriuria, Escherichia coli being the most frequent micro-organism (n=16). CONCLUSION: Preoperative search and management of S. aureus colonization and of bacteriuria in outpatients is possible. Monitoring record must be performed by a member of the hospital staff.

[Effectiveness of systematic investigation for Group B Streptococcus in urine samples to identify colonized pregnant women]. / Impacto de la investigación sistemática de estreptococo del grupo B en orina en la identificación de gestantes colonizadas.

OBJECTIVE: To evaluate the effectiveness of systematic investigation for Group B Streptococcus (GBS) in urine samples to detect colonization in pregnant women. METHODS: This study included 1036 pregnant women whose urine samples were cultured in our laboratory during 2006. Any colony consistent with GBS was identified in urine or in rectovaginal samples submitted for screening of GBS colonization. RESULTS: GBS was recovered in urine samples from 111 of the 1036 women (10.7%), and in 77 of them bacterial count was <10(4) colony forming units/mL. Screening for GBS in rectovaginal samples was performed in 841 of the 925 pregnant women who did not have GBS bacteriuria (10.1% positive results) and in 61 of the 111 with GBS bacteriuria (60.7% positive results; no significant differences were found when results were stratified by colony count). Estimated rectovaginal colonization was 15.4%, and colonization exclusively detected in urine was 4.2%. Only 30% of pregnant women with GBS bacteriuria, but negative antenatal screening cultures who were admitted to our hospital for delivery received intrapartum antibiotic prophylaxis. CONCLUSIONS: Systematic investigation of the presence of GBS in urine samples from pregnant women improves the detection of carriers who are candidates for receiving intrapartum prophylaxis to prevent perinatal GBS infection, when compared with rectovaginal screening culture in the last trimester of gestation alone.

Urinary excretion of pneumococcal cell wall polysaccharide in children.

The urinary excretion of the cell wall polysaccharide of Streptococcus pneumoniae was studied in 92 children with the NOW test. Cell wall polysaccharide was detected in 65% of pneumococcal carriers and in 10% of noncarriers. Excretion rates were similar in healthy children and in children with acute otitis media. The high rate of antigen excretion among nonill carriers suggests that colonization is a major source of urinary antigen in children.

Evaluation of Binax now Streptococcus pneumoniae urinary antigen test in children in a community with a high carriage rate of pneumococcus.

Pneumococcal antigen was present in urine from 49 of 102 well Gambian children. Eighty-nine of the 102 were nasopharyngeal carriers of pneumococci. The positive predictive value for carriage was 96%, and the negative predictive value was 22%. The test is not useful for predicting etiology of disease in populations with a high rate of nasopharyngeal carriage of pneumococci.

Genotyping of Chlamydia trachomatis in urine specimens will facilitate large epidemiological studies.

The serovars of Chlamydia trachomatis-positive urine specimens (n = 81; as detected by PCR and ligase chain reaction) were successfully analyzed in 94% of cases by omp1 PCR-based RFLP analysis. The use of urine specimens and this simple and sensitive typing method will greatly facilitate epidemiological studies of C. trachomatis serovar distribution in asymptomatic C. trachomatis infections in both females and males.

Demonstration of Borrelia burgdorferi DNA in urine samples from healthy humans whose sera contain B. burgdorferi-specific antibodies.

Since the possibility of asymptomatic infection with Borrelia burgdorferi has been suggested by a positive serology found in healthy subjects, we hypothesized that these subjects might excrete borrelial DNA sequences in urine as happens in patients with Lyme borreliosis. We found borrelial sequences by nested PCR in the urine samples from 3 of 13 healthy B. burgdorferi antibody-positive adults but not in urine samples from 79 antibody-negative healthy controls. After therapy with doxycycline, the urine samples were repeatedly negative for B. burgdorferi DNA. We conclude that urinary excretion of borrelial DNA sequences may occur in seropositive healthy subjects during asymptomatic infection. Demonstration of such sequences in urine must be interpreted cautiously and may not necessarily prove a borrelial cause of disease.

Humoral immune responses and cytomegalovirus excretion in children with asymptomatic infection.

Forty-two seropositive children aged 3 to 5 years attending a kindergarten were followed up for 1 year in order to examine the relationship between humoral immunity and cytomegalovirus (CMV) excretion status. Anti-CMV antibodies were measured at the beginning and end of the study by enzyme-linked immunosorbent assay, neutralizing antibody test, and immunoblot techniques. Among these children, 32 persistently shed virus in urine, 2 intermittently shed CMV, and 4 experienced reactivation during the study. Virus was never isolated from 4 seropositive children. The level of anti-CMV IgG antibody in seropositive children who remained nonshedders was significantly higher than in children who shed virus during follow-up. On immunoblots, all seropositive nonshedders reacted to a CMV-specific 65 kD antigen, whereas most shedders (80%) did not. These findings suggest that humoral immunity plays a role in controlling persistent CMV infection in children with asymptomatic infection. However, the humoral immunity measured by the neutralizing test and the presence of antibodies against CMV-specific envelope antigens (116 kD/55 kD) apparently play a limited role in modifying persistent excretion and regulating reactivation of latent CMV. Immune evasion by CMV to block these antigens may explain these results.

Antigenuria in healthy Papua New Guinean children with nasal Haemophilus influenzae type b carriage.

In 100 healthy children under the age of 3 years living in the vicinity of Goroka, Papua New Guinea, the nares were cultured for Haemophilus influenzae type b (Hib), and a urine sample was obtained for measurement of Hib polysaccharide (PS) by ELISA. Hib carriage was detected in nine children and Hib PS was detected in the urine of 11. Hib PS was found in seven of nine Hib nasal carriers compared with four of 91 healthy children without Hib in their nares (p < 0.001). The range of urine antigen concentrations in the two groups was similar (0.6 to 2.7 ng/ml). The relative risk of antigenuria in the carriers, compared with the children with negative nares cultures, was 58 (95% confidence interval, 10.5-324). These data extend previous observations from Hib carriers studied in the United States and show that Hib carriage in children from a developing country is associated with antigenuria. Further studies are needed to determine whether carriers and patients can be differentiated by differences in the magnitude of the concentration of Hib PS excreted in urine.

Evaluation of three immunoassays for detection of Chlamydia trachomatis in urine specimens from asymptomatic males.

The performances of three commercially available immunoassays (Chlamydiazyme/Antibody Blocking Assay [Abbott Diagnostics, Abbott Park, Ill.], IDEIA [Analytab Products, Plainview, N.Y.], and Microtrak EIA [Syva Co. Palo Alto, Calif.]) were evaluated for the detection of Chlamydia trachomatis in urine specimens from asymptomatic males. Assay results were compared with direct specimen immunofluorescence (DFA) analysis of urine sediment (Syva Microtrak; Syva Co.), which was chosen as the study confirmation assay. An overall Chlamydia prevalence of 7% (24 of 340) was found in our study population, with peak incidences occurring in the adolescent (8 of 93 specimens) and young adult (11 of 146 specimens) age groups. Sensitivity and specificity data among the Chlamydiazyme, IDEIA, and Microtrak enzyme immunoassays (EIAs) were determined to be 79.1 and 99%, 91.7 and 98%, and 95.8 and 99%, respectively. The Microtrak EIA and IDEIA products demonstrated sensitivities and specificities equal to or greater than those claimed for urine specimens. The diagnostic accuracies of these assays on asymptomatic subjects, along with the ease of this collection method, suggest a role for these products as screening tools. The sensitivity of the Chlamydiazyme assay was lower than that claimed previously in symptomatic patients, with 5 of 24 positive specimens demonstrating false-negative results. In those cases, centrifugation of the original immunoassay aliquot material and then DFA examination confirmed specimen positivity. Urine immunoassay screening in combination with DFA confirmation (which was chosen because it has antibody epitopic specificity different from that of the primary assay) provides a high degree of diagnostic precision. The use of noninvasive collection methods could result in greater testing compliance among asymptomatic males and, subsequently, could reduce the incidences of both symptomatic and silent chlamydial infections.