A biogeographic 16S rRNA survey of bacterial communities of ureolytic biomineralization from California public restrooms.

Cara Wademan,Matthew Rolston,Andrea Franzetti,Samantha Barnum,Harold Leverenz,Kahui Lim

doi:10.1371/journal.pone.0262425

Abstract

In this study, we examined the total bacterial community associated with ureolytic biomineralization from urine drainage systems. Biomineral samples were obtained from 11 California Department of Transportation public restrooms fitted with waterless, low-flow, or conventional urinals in 2019. Following high throughput 16S rRNA Illumina sequences processed using the DADA2 pipeline, the microbial diversity assessment of 169 biomineral and urine samples resulted in 3,869 reference sequences aggregated as 598 operational taxonomic units (OTUs). Using PERMANOVA testing, we found strong, significant differences between biomineral samples grouped by intrasystem sampling location and urinal type. Biomineral microbial community profiles and alpha diversities differed significantly when controlling for sampling season. Observational statistics revealed that biomineral samples obtained from waterless urinals contained the largest ureC/16S gene copy ratios and were the least diverse urinal type in terms of Shannon indices. Waterless urinal biomineral samples were largely dominated by the Bacilli class (86.1%) compared to low-flow (41.3%) and conventional samples (20.5%), and had the fewest genera that account for less than 2.5% relative abundance per OTU. Our findings are useful for future microbial ecology studies of urine source-separation technologies, as we have established a comparative basis using a large sample size and study area.

Highlights

Nucleotide sequences were deposited at the National Center for Biotechnology Information (NCBI) Sequencing Read Archive (SRA) under the BioProject Accession number PRJNA699694
After the DADA2 pipeline, the microbial diversity assessment of 169 biomineral and liquid samples obtained from 11 different California state-owned rest areas resulted in a total of 3869 reference sequences aggregated as 598 operational taxonomic units (OTUs)
76.5% of the 598 OTUs were classified at the genus level, 92% at the family level, and 100% at the phylum level

Summary

Materials and methods

A 16S rRNA survey of bacterial communities of ureolytic biomineralization ecology study using high throughput sequencing of the 16S rRNA marker. A 16S rRNA survey of bacterial communities of ureolytic biomineralization demonstrated that neither storage time nor storage temperature substantially altered overall communities relative to more than 500 previously examined soil samples [20]. Each 25 mL PCR reaction contained 1 Unit Kapa2G Robust Hot Start Polymerase (Kapa Biosystems), 1.5 mM MgCl2, 0.2 mM final concentration dNTP mix, 0.2 mM final concentration of each primer and 1 uL of DNA for each sample. To understand the effect of geographic distances and environmental variables on the bacterial community within biomineral samples, a Mantel test was performed [32]. Sequencing reads were deposited to NCBI’s sequencing read archives under the BioProject accession number PRJNA699694

Results and discussion

Limitations of the study

33. Alpha Diversity