The urinary microbiota composition remains stable over time and under various storage conditions

Bundgaard-Nielsen, C.; Ammitzboll, N.; Isse, Y. A.; Muqtar, A.; Jensen, A.-M.; Leutscher, P.; Arenholt, L. T. S.; Hagstrom, S.; Sorensen, S.

doi:10.1101/2019.12.13.875187

bioRxiv · 10.1101/2019.12.13.875187

The urinary microbiota composition remains stable over time and under various storage conditions

Bundgaard-Nielsen, C.·Ammitzboll, N.·Isse, Y. A.·Muqtar, A.·Jensen, A.-M.·Leutscher, P.·Arenholt, L. T. S.·Hagstrom, S.·Sorensen, S.

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Abstract

BackgroundNew sensitive techniques have revealed a large population of bacteria in the human urinary tract, challenging the perception of the urine of healthy humans being sterile. While the role of this urinary microbiota is unknown, dysbiosis has been linked to disorders like urgency urinary incontinence and interstitial cystitis. When comparing studies it is crucial to account for possible confounders introduced due to methodological differences. Here we investigated whether storage condition or time of collection, had any impact on the urinary microbial composition. ResultsFor comparison of different storage conditions, urine was collected from five healthy adult female donors, and analyzed by 16S rRNA gene sequencing. Using the same methods, the daily or day-to-day variation in urinary microbiota was investigated in nineteen healthy donors, including four women, five men, five girls, and five boys. With the exception of two male adult donors, none of the tested conditions gave rise to significant differences in alpha and beta diversities between individuals. Conclusion: The composition of the urinary microbiota was found to be highly resilient to changes introduced by storage temperature and duration. In addition, we did not observe any intrapersonal daily or day-to-day variations in microbiota composition in women, girls or boys. Together our study supports flexibility in study design, when conducting urinary microbiota studies. Author summaryThe discovery of bacteria native to the urinary tract in healthy people, a location previously believed to be sterile, has prompted research into the clinical potential of these bacteria. However, methodological weaknesses can significantly influence such studies, and thus development of robust techniques for investigating these bacteria are needed. In the present study, we investigated whether differences in storage following collection, could affect the bacterial composition of urine samples. Next, we investigated if this composition exhibited daily or day-to-day variations. Firstly, we found, that the bacterial composition of urine could be maintained by storage at -80 {degrees}C, -20 {degrees}C, or refrigerated at 4 {degrees}C. Secondly, the bacterial composition of urine remained stable over time. Overall, the results of this study provide information important to study design in future investigations into the clinical implications of urinary bacteria.

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Bundgaard-Nielsen, C., Ammitzboll, N., Isse, Y. A., Muqtar, A., Jensen, A.-M., Leutscher, P., Arenholt, L. T. S., Hagstrom, S., Sorensen, S.. 2019-12-13. The urinary microbiota composition remains stable over time and under various storage conditions. https://doi.org/10.1101/2019.12.13.875187

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