Culture-Dependent and Sequencing Methods Revealed the Absence of a Bacterial Community Residing in the Urine of Healthy Cats.

Andrea Balboni,Lorenza Urbani,Alessia Rizzardi,Federica Serafini,Stefano De Arcangeli,Sofia Segatore,Mara Battilani,Luca Bano,Francesco Dondi,Giovanni Franzo

doi:10.3389/fvets.2020.00438

Abstract

A growing number of studies suggest that the lower urinary tract of humans and dogs can harbor a urinary microbiota. Nevertheless, a certain concern has developed that the microbiota reported could be due to unaccounted contamination, especially in low-biomass samples. The aim of this study was to investigate the bacterial community which populates the urine of healthy cats using two approaches: a culture-dependent approach which consisted of the expanded quantitative urine culture (EQUC) techniques capable of identifying live bacteria not growing in standard urine cultures, and a culture-independent approach which consisted of 16S ribosomal RNA next generation sequencing (16S rRNA NGS) capable of identifying bacterial DNA and exploring microbial diversity with high resolution. To avoid confounding factors of possible bacterial contamination, the urine was sampled using ultrasound-guided cystocentesis, and several sample controls and negative controls were analyzed. The urine sampled from the 10 cats included in the study showed no bacterial growth in the EQUC procedure. Although several reads were successfully originated using 16S rRNA NGS, a comparable pattern was observed between urine samples and the negative control, and no taxa were statistically accepted as non-contaminant. Taken together, the results obtained allowed stating that no viable bacteria were present in the urine of healthy cats without lower urinary tract disease and urinary tract infections, and that the bacterial DNA detected was of contaminant origin.

Highlights

Until a few years ago, in human and veterinary medicine, the dogma that clinical urine specimens of asymptomatic patients are normally sterile was uniformly accepted as a result of the standard urine culture-negative status
The prevailing assumption that clinical urine specimens of asymptomatic patients can be considered sterile was recently questioned in human medicine following emerging evidence claiming that the lower urinary tract could have urinary microbiota [1,2,3, 5, 8]
With the exception of a study reporting the presence of bacterial DNA in the urine of healthy dogs [6], to date, no data on the bacterial communities that populate the urinary tract of domestic animals are available

Summary

Introduction

Until a few years ago, in human and veterinary medicine, the dogma that clinical urine specimens of asymptomatic patients are normally sterile was uniformly accepted as a result of the standard urine culture-negative status. The characterization of the microbial community present in the urinary tract of healthy cats without urologic diseases can help to understand which bacteria are involved in the maintenance of urinary tract homeostasis and help the veterinary practitioner in the diagnosis and treatment of infections. In this prospective, the bacterial community which populates the urine of healthy cats without a history of clinical signs, and clinicopathological and microbiological findings associated with feline lower urinary tract disease (FLUTD) and UTI was investigated using both culture-dependent and culture-independent methods

Objectives

Methods

Results

Conclusion