Basic HTML Version
MERC – DRAFT Test Plan
7
toxigenic
V. cholerae
are calculated by filtration and selection on TCBS agar and enumerated
using species-specific RNA colony blot (500
μ
l to 1 ml) and
ctxA
DNA colony blot (1-10 ml).
Viable toxigenic
V. cholerae
is assayed with a commercial DFA kit specific for serogroup O1
(New Horizons Diagnostics) using monoclonal antibodies tagged with fluorescein isothiocyanate
(FITC) (Hasan et al. 1994).
Sample and Data Management:
We will take advantage of the established SERC ballast water sample labeling and
databases format and structure for this evaluation. Sample-labels and record keeping check-lists
will be generated using SERC protocols, and data will be stored both in existing SERC databases
(servers) and in a MERC repository for analytical data.
Data Analysis:
Although multiple mesocosms, samples, and measures from each tank will be taken, to
avoid pseudo-replication, the unit of replication for statistical analyses is each trial (n = 5 or 6).
We assume that all measures for a single trial provide one estimate of treatment efficacy. Thus,
treatment efficacy for any biological parameter is estimated as changes found before and after
trial (percent reduction), and as the difference in concentration between treated water and IMO
standards. This approach controls for variation due to temporal changes in environmental
conditions.
4. Evaluation Schedule (planned dates based on current plan and may vary)
•
MERC Test Plan for MSI finalized and Evaluation Agreement signed by March 27, 2009
•
MERC evaluation of the MSI systems initiated by March 30, 2009
•
MERC will complete sample analysis and compile data from the evolution by June 2009
•
MERC will distribute a draft report on the performance of the MSI system for review by
the MERC Advisory Board and MSI by August 2009
•
MERC will submit a final report to MPA, MARAD, NOAA and MSI by fall 2009
5. Data Recording, Processing, and Storage
This section describes methods employed during data recording, processing, and storage
to minimize errors and assure high quality analyses.
Documentation and Records:
A variety of data will be acquired and recorded electronically and manually by MERC
partners (CBL/UMCES, SERC, and UM) during this evaluation. Operational information and
results will generally be documented in field/laboratory record books and on the data
sheet/chain-of-custody forms (see below). Copies of these raw data will be transferred to the
MERC office, which will store it permanently along with the rest of the study data.