N-Nitrosodimethylamine Formation from Treatment of Seasonally and Spatially Varying Source Water

N-nitrosodimethylamine (NDMA) is a disinfection by-product (DBP) that has been classified as a probable human carcinogen in multiple risk assessments. NDMA presence in drinking water is widespread and dependent on source water, disinfectant type, precursors, and water treatment strategies. The objectives of this study were to investigate NDMA formation potential in a modeled monochloramine water treatment plant (WTP) fed by seasonally and spatially varying source water; and to optimize DBP precursor removal by combining conventional and additional treatment techniques. After NDMA analysis, it was found that NDMA formation was significantly dependent on source water type and monochloramine contact time (CT); e.g., at 24 h CT, Cork Brook produced 12.2 ng/L NDMA and Bailey Brook produced 4.2 ng/L NDMA, compared with 72 h CT, Cork Brook produced 4.1 ng/L NDMA and Bailey Brook produced 3.4 ng/L NDMA. No correlations were found between traditional DBP precursors such as total organic carbon and total nitrogen, and the formation of NDMA. The laboratory bench-top treatment system was highly effective at removing traditional DBP precursors, highlighting the need for WTPs to alter their current treatment methods to best accommodate the complex system of DBP control

Characterization of the Microbiome at the World’s Largest Potable Water Reuse Facility

Conventional water resources are not sufficient in many regions to meet the needs of growing populations. Due to cyclical weather cycles, drought, and climate change, water stress has increased worldwide including in Southern California, which serves as a model for regions that integrate reuse of wastewater for both potable and non-potable use. The Orange County Water District (OCWD) Advanced Water Purification Facility (AWPF) is a highly engineered system designed to treat and produce up to 100 million gallons per day (MGD) of purified water from a municipal wastewater source for potable reuse. Routine facility microbial water quality analysis is limited to standard indicators at this and similar facilities. Given recent advances in high throughput DNA sequencing techniques, complete microbial profiling of communities in water samples is now possible. By using 16S/18S rRNA gene sequencing, metagenomic and metatranscriptomic sequencing coupled to a highly accurate identification method along with 16S rRNA gene qPCR, we describe a detailed view of the total microbial community throughout the facility. The total bacterial load of the water at stages of the treatment train ranged from 3.02 × 106 copies in source, unchlorinated wastewater feed to 5.49 × 101 copies of 16S rRNA gene/mL after treatment (consisting of microfiltration, reverse osmosis, and ultraviolet/advanced oxidation). Microbial diversity and load decreased by several orders of magnitude after microfiltration and reverse osmosis treatment, falling to almost non-detectable levels that more closely resembled controls of molecular grade laboratory water than the biomass detected in the source water. The presence of antibiotic resistance genes and viruses was also greatly reduced. Overall, system design performance was achieved, and comprehensive microbial community analysis was found to enable a more complete characterization of the water/wastewater microbial signature

Microbial resolution of whole genome shotgun and 16S amplicon metagenomic sequencing using publicly available NEON data.

Microorganisms are ubiquitous in the biosphere, playing a crucial role in both biogeochemistry of the planet and human health. However, identifying these microorganisms and defining their function are challenging. Widely used approaches in comparative metagenomics, 16S amplicon sequencing and whole genome shotgun sequencing (WGS), have provided access to DNA sequencing analysis to identify microorganisms and evaluate diversity and abundance in various environments. However, advances in parallel high-throughput DNA sequencing in the past decade have introduced major hurdles, namely standardization of methods, data storage, reproducible interoperability of results, and data sharing. The National Ecological Observatory Network (NEON), established by the National Science Foundation, enables all researchers to address queries on a regional to continental scale around a variety of environmental challenges and provide high-quality, integrated, and standardized data from field sites across the U.S. As the amount of metagenomic data continues to grow, standardized procedures that allow results across projects to be assessed and compared is becoming increasingly important in the field of metagenomics. We demonstrate the feasibility of using publicly available NEON soil metagenomic sequencing datasets in combination with open access Metagenomics Rapid Annotation using the Subsystem Technology (MG-RAST) server to illustrate advantages of WGS compared to 16S amplicon sequencing. Four WGS and four 16S amplicon sequence datasets, from surface soil samples prepared by NEON investigators, were selected for comparison, using standardized protocols collected at the same locations in Colorado between April-July 2014. The dominant bacterial phyla detected across samples agreed between sequencing methodologies. However, WGS yielded greater microbial resolution, increased accuracy, and allowed identification of more genera of bacteria, archaea, viruses, and eukaryota, and putative functional genes that would have gone undetected using 16S amplicon sequencing. NEON open data will be useful for future studies characterizing and quantifying complex ecological processes associated with changing aquatic and terrestrial ecosystems

N-Nitrosodimethylamine Formation from Treatment of Seasonally and Spatially Varying Source Water

N-nitrosodimethylamine (NDMA) is a disinfection by-product (DBP) that has been classified as a probable human carcinogen in multiple risk assessments. NDMA presence in drinking water is widespread and dependent on source water, disinfectant type, precursors, and water treatment strategies. The objectives of this study were to investigate NDMA formation potential in a modeled monochloramine water treatment plant (WTP) fed by seasonally and spatially varying source water; and to optimize DBP precursor removal by combining conventional and additional treatment techniques. After NDMA analysis, it was found that NDMA formation was significantly dependent on source water type and monochloramine contact time (CT); e.g., at 24 h CT, Cork Brook produced 12.2 ng/L NDMA and Bailey Brook produced 4.2 ng/L NDMA, compared with 72 h CT, Cork Brook produced 4.1 ng/L NDMA and Bailey Brook produced 3.4 ng/L NDMA. No correlations were found between traditional DBP precursors such as total organic carbon and total nitrogen, and the formation of NDMA. The laboratory bench-top treatment system was highly effective at removing traditional DBP precursors, highlighting the need for WTPs to alter their current treatment methods to best accommodate the complex system of DBP control

A comparative analysis of drinking water employing metagenomics.

The microbiological content of drinking water traditionally is determined by employing culture-dependent methods that are unable to detect all microorganisms, especially those that are not culturable. High-throughput sequencing now makes it possible to determine the microbiome of drinking water. Thus, the natural microbiota of water and water distribution systems can now be determined more accurately and analyzed in significantly greater detail, providing comprehensive understanding of the microbial community of drinking water applicable to public health. In this study, shotgun metagenomic analysis was performed to determine the microbiological content of drinking water and to provide a preliminary assessment of tap, drinking fountain, sparkling natural mineral, and non-mineral bottled water. Predominant bacterial species detected were members of the phyla Actinobacteria and Proteobacteria, notably the genera Alishewanella, Salmonella, and Propionibacterium in non-carbonated non-mineral bottled water, Methyloversatilis and Methylibium in sparkling natural mineral water, and Mycobacterium and Afipia in tap and drinking fountain water. Fecal indicator bacteria, i.e., Escherichia coli or enterococci, were not detected in any samples examined in this study. Bacteriophages and DNA encoding a few virulence-associated factors were detected but determined to be present only at low abundance. Antibiotic resistance markers were detected only at abundance values below our threshold of confidence. DNA of opportunistic plant and animal pathogens was identified in some samples and these included bacteria (Mycobacterium spp.), protozoa (Acanthamoeba mauritaniensis and Acanthamoeba palestinensis), and fungi (Melampsora pinitorqua and Chryosporium queenslandicum). Archaeal DNA (Candidatus Nitrosoarchaeum) was detected only in sparkling natural mineral water. This preliminary study reports the complete microbiome (bacteria, viruses, fungi, and protists) of selected types of drinking water employing whole-genome high-throughput sequencing and bioinformatics. Investigation into activity and function of the organisms detected is in progress