Rapid Fluorescent Assays for Coliforms in Shipboard Wastewater

One challenge encountered aboard marine vessels is the treatment, storage, and discharge of wastewater from marine sanitation devices (MSDs). Improperly treated waste water has a high potential for transmitting diseases and harming the ecosystem. The presence of Escherichia coli is routinely used as a key indicator of poor wastewater treatment. Conventional E. coli assays, which are required to detect fewer than 100 bacteria/ml, are laborious selective culture plating methods and require 24-48 hours to complete, increasing onboard storage and sanitation requirements. During Phase I, Luna"s rapid fluorescent viability assays, previously developed for monitoring biothreat bacteria, will be adapted for use with E coli and non-fecal coliforms. To rapidly predict the efficacy of MSDs, we will develop and validate high-throughput syringe formats for detection of coliforms by enzyme hydrolysis of fluorescent substrates. Luna Innovations proposes to i) down select fluorescent assays, ii) optimize enzyme induction, iii) develop a rapid high-through syringe filter format and iv) demonstrate the fluorescent format using fresh MSD wastewater effluents. During Phase II, Luna will i) validate the methods, ii) perform full scale evaluations on Naval vessels, iii) explore potential sources for Phase III support and iv) begin transition to NAVSEA for acquisition and commercialization.