Longitudinal variability in the urinary microbiota of healthy premenopausal women and the relation to neighboring microbial communities: A pilot study

Background The understanding of longitudinal changes in the urinary microbiota of healthy women and its relation to intestinal microbiota is limited. Methods From a cohort of 15 premenopausal women without known urogenital disease or current symptoms, we collected catheter urine (CU), vaginal and periurethral swabs, and fecal samples on four visits over six months. Additionally, ten participants provided CU and midstream urine (MU) to assess comparability. Urine was subjected to expanded culture. 16S rRNA gene sequencing was performed on all urine, fecal, and selected vaginal and periurethral samples. Sequence reads were processed (DADA2 pipeline) and analyzed using QIIME 2 and R. Results Relative abundances of urinary microbiota were variable over 6–18 months. The degree of intraindividual variability of urinary microbiota was higher than that found in fecal samples. Still, nearly half of the observed beta diversity of all urine samples could be attributed to differences between volunteers (R2 = 0.48, p = 0.001). After stratification by volunteer, time since last sexual intercourse was shown to be a factor significantly contributing to beta diversity (R2 = 0.14, p = 0.001). We observed a close relatedness of urogenital microbial habitats and a clear distinction from intestinal microbiota in the overall betadiversity analysis. Microbiota compositions derived from MU differed only slightly from CU compositions. Within this analysis of low-biomass samples, we identified contaminating sequences potentially stemming from sequencing reagents. Conclusions Results from our longitudinal cohort study confirmed the presence of a rather variable individual urinary microbiota in premenopausal women. These findings from catheter urine complement previous observations on temporal dynamics in voided urine. The higher intraindividual variability of urinary microbiota as compared to fecal microbiota will be a challenge for future studies investigating associations with urogenital diseases and aiming at identifying pathogenic microbiota signatures.

Antimicrobial Susceptibility Pattern and Molecular Identification of Acinetobacter baumannii in Alex Ekwueme-Federal University Teaching Hospital Abakaliki, Nigeria

Background and Objectives: Acinetobacter baumannii, a notorious opportunistic pathogen known to seriously affect debilitated individuals especially intensive care unit (ICU) patients and others with underlying illness, have consistently jeopardized many antibiotics. This study was therefore aimed to ascertain the antimicrobial susceptibility profile and molecularly identify A. baumannii pathogens in Alex Ekwueme-Federal University Teaching Hospital, Abakaliki, Nigeria. Methodology: A total of 385 clinical samples were collected aseptically from debilitated patients and analyzed following standard microbiological procedures. Acinetobacter species was confirmed by Gram staining reaction and biochemical tests. All bacterial isolates were phenotypically screened for multidrug resistance using the Kirby–Bauer disc diffusion technique and results interpreted as per CLSI criteria. A. baumannii isolates were finally confirmed using 16S rRNA sequencing. Results: A total of 23(6%) A. baumannii isolates were recovered from 385 clinical samples collected from 87 patients comprising 48 males and 39 females admitted in various hospital wards of AE-FETHA. The age of the patients varied from 20–79 years. The commonest sites for isolation of A. baumannii pathogen were catheter urine (8/8%) and wound sores (7/8%). The highest percentage resistance was observed with cefuroxime (96%), tetracycline (96%), sulfamethoxazole/trimethoprim (96 %), and ofloxacin (91%) while meropenem (91%) and imipenem (78%) were the most effective antibiotics against A. baumannii. The isolated A. baumannii was re-confirmed genotypically by 16S rRNA gene amplification. Variations were observed in the gene sequence of all the isolated A. baumannii. Conclusion: Catheter urine, wound sores, and respiratory fluids were the more easily colonized samples. Also, high frequency of multidrug resistance observed in this study further established A. baumannii as a notorious opportunistic pathogen.

The Origin of Tumor DNA in Urine of Urogenital Cancer Patients: Local Shedding and Transrenal Excretion

In urogenital cancers, urine as a liquid biopsy for non-invasive cancer detection holds great promise for future clinical application. Their anatomical position allows for the local shedding of tumor DNA, but recent data indicate that tumor DNA in urine might also result from transrenal excretion. This study aims to assess the origin of tumor-associated DNA in the urine of 5 bladder and 25 cervical cancer patients. Besides natural voided urine, paired urine samples were collected in which contact with the local tumor was circumvented to bypass local shedding. The latter concerned nephrostomy urine in bladder cancer patients, and catheter urine in cervical cancer patients. Methylation levels of GHSR, SST, and ZIC1 were determined using paired bladder tumor tissues and cervical scrapes as a reference. Urinary methylation levels were compared to natural voided urine of matched controls. To support methylation results, mutation analysis was performed in urine and tissue samples of bladder cancer patients. Increased methylation levels were not only found in natural voided urine from bladder and cervical cancer patients, but also in the corresponding nephrostomy and catheter urine. DNA mutations detected in bladder tumor tissues were also detectable in all paired natural voided urine as well as in a subset of nephrostomy urine. These results provide the first evidence that the suitability of urine as a liquid biopsy for urogenital cancers relies both on the local shedding of tumor cells and cell fragments, as well as the transrenal excretion of tumor DNA into the urine.

Enterococcus faecalis Polymicrobial Interactions Facilitate Biofilm Formation, Antibiotic Recalcitrance, and Persistent Colonization of the Catheterized Urinary Tract

Indwelling urinary catheters are common in health care settings and can lead to catheter-associated urinary tract infection (CAUTI). Long-term catheterization causes polymicrobial colonization of the catheter and urine, for which the clinical significance is poorly understood. Through prospective assessment of catheter urine colonization, we identified Enterococcus faecalis and Proteus mirabilis as the most prevalent and persistent co-colonizers. Clinical isolates of both species successfully co-colonized in a murine model of CAUTI, and they were observed to co-localize on catheter biofilms during infection. We further demonstrate that P. mirabilis preferentially adheres to E. faecalis during biofilm formation, and that contact-dependent interactions between E. faecalis and P. mirabilis facilitate establishment of a robust biofilm architecture that enhances antimicrobial resistance for both species. E. faecalis may therefore act as a pioneer species on urinary catheters, establishing an ideal surface for persistent colonization by more traditional pathogens such as P. mirabilis.

Enterococcus faecalis Polymicrobial Interactions Facilitate Biofilm Formation, Antibiotic Recalcitrance, and Persistent Colonization of the Catheterized Urinary Tract

Indwelling urinary catheters are common in healthcare settings and can lead to catheter-associated urinary tract infection (CAUTI). Long-term catheterization causes polymicrobial colonization of the catheter and urine, for which the clinical significance is poorly understood. Through prospective assessment of catheter urine colonization, we identified Enterococcus faecalis and Proteus mirabilis as the most prevalent and persistent co-colonizers. Clinical isolates of both species successfully co-colonized in a murine model of CAUTI, and they were observed to co-localize on catheter biofilms during infection. We further demonstrate that P. mirabilis preferentially adheres to E. faecalis during biofilm formation, and that contact-dependent interactions between E. faecalis and P. mirabilis facilitate establishment of a robust biofilm architecture that enhances antimicrobial resistance for both species. E. faecalis may therefore act as a pioneer species on urinary catheters, establishing an ideal surface for persistent colonization by more traditional pathogens such as P. mirabilis.

Urine Output Measurement by a Novel Electronic Urinometer is much more Accurate than by Conventional Urinometer

Introduction: Assessment of urine output in hospitalized patients or those undergoing complex surgery is essential for preventing Acute Kidney Injury and increased mortality. Currently, measurements of urine output are made by visually inspecting the volume of urine that accumulates in a bag attached to a catheter which resides in the bladder by way of a manual urinometer. Several electronic urinometers have been tested but have not been clinically proven or accepted. We report preliminary results of a novel Electronic Urinometer (EU) and compared its accuracy to a conventional Manual Urinometer (MU). Methods: The study included 22 consecutive ICU patients whose urine output was collected with a foley catheter. Urine output was measured for all patients by EU as well as by MU. Both measurements were compared to analysis of the volume in a Graduated Cylinder (GC). Results: The EU was highly accurate in measurement of both urine volume and urine flow with an average deviation from GC 4 ± 3.0% vs 17 ± 23% for the MU (P<0.01). Conclusion: This study showed that measuring urine output in ICU patients by an EU is more accurate than conventional MU.

Short communication: Evaluation of tubal patency in repeat breeder Holstein cows

Aim of study: To evaluate tubal patency in repeat breeder Holstein cows with the purpose of estimating the frequency of oviduct obstruction as a cause of the repeat breeding syndrome.Area of study: Lugo (NW Spain)Material and methods: In 50 repeat breeding cows, a solution of the dye phenol-sulphon-phthalein (PSP) was instilled into one uterine horn with the aid of a balloon catheter. Urine was collected 15, 25 and 45 min post-PSP infusion. Urine of a reddish color was used as indicator of tubal patency, as the PSP had passed across the oviduct into the abdominal cavity and was eliminated in urine. The other oviduct was evaluated on a different day.Main results: Of the 50 cows tested, 44% had some degree of oviduct obstruction: 4% had bilateral oviduct occlusion, 20% unilateral oviduct occlusion, 16% bilateral oviduct stenosis and 4% unilateral oviduct stenosis. No significant relationship could be established between tubal impermeability and postpartum reproductive or metabolic diseases.Research highlights: Tubal stenosis or occlusions were frequent and they are likely to contribute to the repeat breeding syndrome in dairy cows; thus, tubal patency should be routinely evaluated in repeat breeders. The PSP test is easy to perform, well tolerated by animals, and allows evaluating both oviducts separately.

2569. High Incidence of Enterovirus, HHV6, Parechovirus and Adenovirus Blood Viremia in Children 0 to 3 Years Old Presenting With Fever Without Source

Background Fever without source (FWS) is defined as a fever in which an extensive history and clinical examination fail to identify a cause. Although the vast majority of children with FWS have a self-limited viral infection, up to 10–25% have a serious bacterial infection (SBI). Therefore, many children require invasive diagnostic tests, hospital admission, and empirical administration of broad-spectrum antibiotics. The aim of this study was to assess the respective role of Human enterovirus (HEV), human parechovirus (HPeV), adenovirus (ADV) and herpesvirus type 6 (HHV6) viremia in children &lt;3 years old presenting with FWS. Methods Prospective monocentric diagnostic study. Between November 2015 to December 2017, children &lt;3 year olds with FWS had, in addition to the standardized institutional work-up for FWS, plasma tested by real-time (reverse-transcription) polymerase chain reaction (PCR) for ADV, HHV6, HEV, and HPeV. Specimens with cycle threshold values &lt;40 were considered positive. Quantification was performed on positive specimens for HEV, ADV, and HHV6 specimens when volume permitted. Results One hundred thirty-five patients had plasma PCR for ADV, HHV6, HEV, and HPeV. Male:female ratio was 1.45:1 and median age was 2.4 months (interquartile range 1.3–9.7). Among those, 47/135 (34.8%) had at least 1 virus detected in the plasma. More specifically, HEV was detected in 19 patients (14.1%), HHV6 in 15 (11.1%), HPeV in 8 (5.9%), and ADV in 7 (5.2%). Co-infection with 2 viruses was detected in 2 patients (ADV/HEV and ADV/HPeV). No patient with positive plasma PCR had a positive blood or CSF culture. Two patients with positive plasma PCR fulfilled American Academy of Pediatrics criteria for urinary tract infection. The first was HEV+ in plasma and CSF, midstream urine was positive for leukocytes and grew E. coli 106 CFU/mL, whereas the second was HHV6+ in plasma and catheter urine was positive leukocytes/nitrites and grew P. mirabilis 105 CFU/mL. Conclusion This epidemiological study highlights the frequent detection of active enteroviral, adenoviral, and HHV6 infections in plasma of children with FWS. Virus–virus and virus–bacteria co-infections are rare. Further studies are needed to establish causality between FWS and viremia. Disclosures A. G. L’Huillier, Gertrude Von Meissner foundation: Investigator, Research grant. Ernst and Lucie Schmidheiny foundation: Investigator, Research grant. Research and Development Grant, Geneva University Hospitals: Investigator, Research grant. L. Kaiser, Swiss National Funds: Investigator, Research grant. A. Galetto-Lacour, Gertrude Von Meissner foundation: Investigator, Research grant. Ernst and Lucie Schmidheiny foundation: Investigator, Research grant. Research and Development Grant, Geneva University Hospitals: Investigator, Research grant.

Resistensi Antibakteri pada Pasien Infeksi Saluran Kemih (ISK) dengan Kateterisasi Urin di Bagian Penyakit Dalam RSUD Arifin Achmad Pekanbaru

Uninary tract infection (UTI) is a common health problem among the people in the world, including Indonesia.Approximately 80% of UTI in hospital are ascociated with urethral catheter used in hospitalized patients. The increaseof catheter- related UTI is olso followed by increase of antibiotic used. The diffrence in distribution of UTI causingbacteria and antibiotics resistance associated with cathetetization can change according to time and place. This stdyaims to find the pattern of antibiotic resistance in the urine of catheter related UTI patient. The research was done inDepartement of Internal Medicine RSUD arifin Achmad Pekanbaru. 31 samples were collected from hospitalizedpatient who used catheter at last 3 days and taken with indwelling catheter urine. Culture, colony count, colonyidentification and resistance test was performed by Kirby Bauer method. The result was interpretated based onClinical and Laboratory Standard Institude (CLSI). UTI found in 19 samples (61.29%), 9 samples founs no bacteria(29.03%) and Candida albicans on 3 samples (9.68%). UTI caused mostly by Gram-negative bacteria (57.89%),Gram- positive bacteria found in 42.11% of samples. The highest resistance (100%) was found on cefotaxim, cefriaxon,cefalexin and tertracyclin. The lowest resistance (73.68%) found on merofenem and co-trimoxazole.

INCIDENCES OF CANDIDA ALBICANS AND NON-ALBICANS AMONG CATHETER-ASSOCIATED URINARY TRACT INFECTION PATIENTS OF AKOLA CITY

Objective: Candida spp. is the third leading cause of catheter-related infections. Candida species is a part of human microflora and it becomes pathogenic when certain conditions are present and cause an opportunistic infections. The present study was undertaken to determine incidences of Candida albicans and non-albicans among catheterized urinary tract infection (UTI) patients of Akola city.Methods: A total 60 catheter urine samples were collected from patient of all the age group and both sex who had indwelling urinary catheter. The collected catheterized urine samples of patients from various hospitals of Akola city were used for isolation using HiCrome Candida differential agar.Results: It was found that highest frequency of isolation of Candida spp. was from age group 61-70 years. The predominance of male candidate was more than female having Candida spp. in catheter-associated UTI (C-UTI). Among the Candida spp. C. albicans (64.81%) was predominant over non-albicans spp. while in non-albicans Candida krusei and Candida glabrata were predominant showing 11.11% incidences. It was followed by Candida tropicalis (9.2%) and Candida parapsilosis (3.7%).Conclusion: The incidences of C. albicans and non-albicans were high among catheter-associated UTI patients.