Elucidation of Taste- and Odor-Producing Bacteria and Toxigenic Cyanobacteria in a Midwestern Drinking Water Supply Reservoir by Shotgun Metagenomic Analysis

Timothy G. Otten, Jennifer L. Graham, Theodore D. Harris, Theo W. Dreher

While commonplace in clinical settings, DNA based assays for identification or enumeration of drinking water pathogens and other biological contaminants remain widely unadopted by the monitoring community. In this study, shotgun metagenomics was used to identify taste-and-odor producers and toxin-producing cyanobacteria over a 2-year period in a drinking water reservoir. The sequencing data implicated several cyanobacteria, including: Anabaena sp., Microcystis sp. and an unresolved member of the family Oscillatoriales as the likely principal producers of geosmin, microcystin and 2-methylisoborneol (MIB), respectively. To further demonstrate this, quantitative PCR assays targeting geosmin-producing Anabaena and microcystin-producing Microcystis were utilized, and these data were fitted using generalized linear models and compared with routine monitoring data including: microscopic cell counts, sonde-based physicochemical analyses and assays of all inorganic and organic nitrogen and phosphorus forms and fractions. The QPCR assays explained the greatest variation in observed geosmin (Adj. R(2)=0.71) and microcystin (Adj. R(2)=0.84) concentrations over the study period, highlighting their potential for routine monitoring applications. The origin of the monoterpene cyclase required for MIB biosynthesis was putatively linked to a periphytic cyanobacterial mat attached to the concrete drinking water inflow structure. We conclude that shotgun metagenomics can be used to identify microbial agents involved in water quality deterioration and to guide PCR assay selection or design for routine monitoring purposes. Finally, we offer estimates of microbial diversity and metagenomic coverage of our datasets for reference to others wishing to apply shotgun metagenomics to other lacustrine systems. Cyanobacterial toxins and microbial taste-and-odor compounds are a growing concern for drinking water utilities reliant upon surface water resources. Specific identification of the microorganism(s) responsible for water quality degradation is often complicated by the presence of co-occurring taxa capable of producing these undesirable metabolites. Here we present a framework for how shotgun metagenomics can be used to definitively identify problematic microorganisms, and how these data can guide the development of rapid genetic assays for routine monitoring purposes.