Ram Lab GLPF-funded work to be presented at IAGLR, June 2013

The Ram Laboratory will present recent results from their Great Lakes Protection Fund supported project on "Automated Ballast Verification" at the upcoming conference of the International Association of Great Lakes Research (IAGLR). The content of the abstract is as follows:

AKRAM, A.C., MONIRI JAVID, R., SINGH, S.B., REED, E.A., GIZICKI, J.P., NOMAN, S., BASU, A.S. and
RAM, J.L., Wayne State University, Detroit, MI, 48201. Automated Testing Device for Live-Dead Analysis
of Ballast Water Organisms.
Methods for verifying ballast water treatments are needed to protect the Great Lakes from discharge of live
non-native organisms or pathogens. Here we describe prototypes for automated viability testing using
fluorescein diacetate (FDA), a membrane permeable fluorogen, to differentiate live from dead bacteria and
algae. An automated system captured lab cultured, environmental, or ballast organisms on 0.2 um filters,
backwashed them with buffer and FDA for subsequent fluorescence measurements, and washed the filters
with sterile water for serial automated reuse. Preliminary manual versions of these procedures were also
tested. Fluorescence in the presence of live organisms increased linearly over time and decreased linearly
with sample dilutions (r2=0.92). After plankton and ballast water treatment with heat or chlorine,
fluorescence was greatly decreased, to levels near control (sterile water). Cost reductions in the detection
system included changing from fluorescence plate reader (>$20,000), to a fiber optics spectrometer
(~$5,000), to recent electronic prototypes costing <$500. Automated measurements of FDA hydrolysis with
a reusable filter backwash system under computer control should be applicable to real-time remotecontrolled
monitoring of live organisms in ballast water (patent pending).