Development of an automated ballast water treatment verification system utilizing fluorescein diacetate hydrolysis as a measure of treatment efficacy

• Published in: Water research (Oxford) Vol. 70; pp. 404 - 413
• Main Authors: Akram, A.C., Noman, S., Moniri-Javid, R., Gizicki, J.P., Reed, E.A., Singh, S.B., Basu, A.S., Banno, F., Fujimoto, M., Ram, J.L.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.03.2015
• Subjects: Automated; Automation; Bacteria; Bacteria - isolation & purification; Ballast; Ballast verification; Buffers; Environmental Monitoring - methods; Fluorescein diacetate; Fluoresceins - chemistry; Fluorescence; Fresh Water - analysis; Fresh Water - microbiology; Hydrolysis; Organisms; Ships; Simulation; Viability testing; Waste Disposal, Fluid; Water Purification - standards; Water treatment; North America, Detroit R; North America, Great Lakes; Ballast verification; Fluorescein diacetate; Automation; Viability testing
• ISSN: 0043-1354; 1879-2448
• DOI: 10.1016/j.watres.2014.12.009

Methods for verifying ballast water treatments in foreign vessels are needed to protect the Great Lakes from the discharge of live non-native organisms or pathogens. A prototype automated viability test system using fluorescein diacetate (FDA), a membrane permeable fluorogen, to differentiate live from dead bacteria and algae is described. The automated fluorescence intensity detection device (AFIDD) captures cultured algae or organisms in Detroit River water (simulated ballast water) on 0.2 μm filters, backwashes them from the filter into a cuvette with buffer and FDA for subsequent fluorescence intensity measurements, and washes the filters with sterile water for serial automated reuse. Preliminary manual versions of these procedures were also tested. Tests of various buffers determined N,N-Bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, N,N-Bis(2-hydroxyethyl)taurine (BES) and 3-(N-morpholino)propanesulfonic acid (MOPS) at pH 7.0 to be the best buffers, causing the least spontaneous FDA breakdown without inhibiting enzymatic activity. Fluorescence in the presence of live organisms increased linearly over time, and the rate of increase was dependent on the sample concentration. Following simulated ballast water treatments with heat or chlorine, the fluorescence produced by Detroit River samples decreased to near control (sterile water) levels. Automated measurements of FDA hydrolysis with a reusable filter backwash system should be applicable to near real-time remote-controlled monitoring of live organisms in ballast water. [Display omitted] •Fluorescein released from fluorescein diacetate detects viable organisms.•Filter concentration and backwash enhances the viable organism signal.•AFIDD automates water sample handling and fluorescence measurements.•Rapid automated verification of simulated ballast treatments is demonstrated.