Source-separated human urine was collected from six public events to study the impact of urine processing and storage on bacterial community composition and viability. Illumina 16S rRNA gene sequencing revealed a complex community of bacteria in fresh urine that differed across collection events. Despite the harsh chemical conditions of stored urine (pH > 9 and total ammonia nitrogen > 4000 mg N/L), bacteria consistently grew to 5 ± 2 × 10 cells/mL. Storing hydrolyzed urine for any amount of time significantly reduced the number of operational taxonomic units (OTUs) to 130 ± 70, increased Pielou evenness to 0.60 ± 0.06, and produced communities dominated by Clostridiales and Lactobacillales. After 80 days of storage, all six urine samples from different starting materials converged to these characteristics. Urine pasteurization or struvite precipitation did not change the microbial community, even when pasteurized urine was stored for an additional 70 days. Pasteurization decreased metabolic activity by 50 ± 10% and additional storage after pasteurization did not lead to recovery of metabolic activity. Urine-derived fertilizers consistently contained 16S rRNA genes belonging to Tissierella, Erysipelothrix, Atopostipes, Bacteroides, and many Clostridiales OTUs; additional experiments must determine whether pathogenic species are present, responsible for observed metabolic activity, or regrow when applied.

A low-cost tap water fingerprinting technique was evaluated using the coffee-ring effect, a phenomenon by which tap water droplets leave distinguishable "fingerprint" residue patterns after water evaporates. Tap waters from communities across southern Michigan dried on aluminum and photographed with a cell phone camera and 30× loupe produced unique and reproducible images. A convolutional neural network (CNN) model was trained using the images from the Michigan tap waters, and despite the small size of the image dataset, the model assigned images into groups with similar water chemistry with 80% accuracy. Synthetic solutions containing only the majority species measured in Detroit, Lansing, and Michigan State University tap waters did not display the same residue patterns as collected waters; thus, the lower concentration species also influence the tap water "fingerprint". Residue pattern images from salt mixtures with an array of sodium, calcium, magnesium, chloride, bicarbonate, and sulfate concentrations were analyzed by measuring features observed in the photographs as well as using principal component analysis (PCA) on the image files and particles measurements. These analyses together highlighted differences in the residue patterns associated with the water chemistry in the sample. The results of these experiments suggest that the unique and reproducible residue patterns of tap water samples that can be imaged with a cell phone camera and a loupe contain a wealth of information about the overall composition of the tap water, and thus, the phenomenon should be further explored for potential use in lowcost tap water fingerprinting. † Data and CNN model are available at https://doi.org/10.5281/zenodo.3550247 ‡ Electronic supplementary information (ESI) available: Replicate images, synthetic tap water recipes, a trilinear plot for the collected tap waters, and detailed descriptions of residue patterns according to water chemistry. See

Source separation of urine is a novel strategy that facilitates improved nutrient recovery and micropollutant management. The Rich Earth Institute operates the United States' first regional urine recycling program, collecting source-separated urine from households and producing a sanitized fertilizer product for use by local farmers. The purpose of this program is to provide practical experience and quantitative data on all stages of the urine recycling process, and to create a platform to allow detailed research into specific aspects of the process in a real-world context. Current research topics at the Institute include the fate of pharmaceutical and biological constituents when urine is used as fertilizer, the effect on crop yield of urine fertilizer in comparison with synthetic fertilizer, and methods for transforming and concentrating urine to reduce the cost of storage and transport.