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Bacterial DNA and live bacteria have been detected in human urine in the absence of clinical infection, challenging the prevailing dogma that urine is normally sterile. Urgency urinary incontinence (UUI) is a poorly understood urinary condition characterized by symptoms that overlap urinary infection, including urinary urgency and increased frequency with urinary incontinence. The recent discovery of the urinary microbiome warrants investigation into whether bacteria contribute to UUI. In this study, we used 16S rRNA gene sequencing to classify bacterial DNA and expanded quantitative urine culture (EQUC) techniques to isolate live bacteria in urine collected by using a transurethral catheter from women with UUI and, in comparison, a cohort without UUI. For these cohorts, we demonstrated that the UUI and non-UUI urinary microbiomes differ by group based on both sequence and culture evidences. Compared to the non-UUI microbiome, sequencing experiments revealed that the UUI microbiome was composed of increased Gardnerella and decreased Lactobacillus. Nine genera (Actinobaculum, Actinomyces, Aerococcus, Arthrobacter, Corynebacterium, Gardnerella, Oligella, Staphylococcus, and Streptococcus) were more frequently cultured from the UUI cohort. Although Lactobacillus was isolated from both cohorts, distinctions existed at the species level, with Lactobacillus gasseri detected more frequently in the UUI cohort and Lactobacillus crispatus most frequently detected in controls. Combined, these data suggest that potentially important differences exist in the urinary microbiomes of women with and without UUI, which have strong implications in prevention, diagnosis, or treatment of UUI.

Our previous study showed that bacterial genomes can be identified using 16S rRNA sequencing in urine specimens of both symptomatic and asymptomatic patients who are culture negative according to standard urine culture protocols. In the present study, we used a modified culture protocol that included plating larger volumes of urine, incubation under varied atmospheric conditions, and prolonged incubation times to demonstrate that many of the organisms identified in urine by 16S rRNA gene sequencing are, in fact, cultivable using an expanded quantitative urine culture (EQUC) protocol. Sixty-five urine specimens (from 41 patients with overactive bladder and 24 controls) were examined using both the standard and EQUC culture techniques. Fifty-two of the 65 urine samples (80%) grew bacterial species using EQUC, while the majority of these (48/52 [92%]) were reported as no growth at 10 3 CFU/ml by the clinical microbiology laboratory using the standard urine culture protocol. Thirty-five different genera and 85 different species were identified by EQUC. The most prevalent genera isolated were Lactobacillus (15%), followed by Corynebacterium (14.2%), Streptococcus (11.9%), Actinomyces (6.9%), and Staphylococcus (6.9%). Other genera commonly isolated include Aerococcus, Gardnerella, Bifidobacterium, and Actinobaculum. Our current study demonstrates that urine contains communities of living bacteria that comprise a resident female urine microbiota. Overactive bladder (OAB) is a highly prevalent syndrome characterized by urinary urgency with or without urge urinary incontinence and is often associated with frequency and nocturia (1). The etiology of OAB is often unclear and antimuscarinic treatments aimed at relaxing the bladder are ineffective in a large percentage of OAB sufferers, thereby suggesting etiologies outside neuromuscular dysfunction (2). One possibility is that OAB symptoms are influenced by microbes that inhabit the lower urinary tract (urinary microbiota).The microbiota of the female urinary tract has been poorly described; primarily, because a "culture-negative" status has been equated with the dogma that normal urine is sterile. Yet, emerging evidence indicates that the lower urinary tract can have a urinary microbiota (3-8). For example, our group previously reported the use of 16S rRNA gene sequencing to identify bacterial DNA (urinary microbiome) in culture-negative urine specimens collected from women diagnosed with pelvic prolapse and/or urinary incontinence, as well as from urine of women without urinary symptoms (4). Other investigators also have used culture-independent 16S rRNA gene sequencing to obtain evidence of diverse bacteria that are not routinely cultivated by clinical microbiology laboratories in the urine of both women and men (3, 6, 9, 10).Most of our previously sequenced urine specimens underwent standard clinical urine cultures that were reported as "no growth" at a 1:1000 dilution by our diagnostic microbiology laboratory (4). On the basis of this sequence-based evidence, which su...