Here, we describe the methodologies adopted to ensure that natural seawater, used as "influent water" for the land test, complies with the requirement that should be fulfilled to show the efficacy of the new ballast water treatment system (BWTS). The new BWTS was located on the coast of SW Sicily (Italy), and the sampled seawater showed that bacteria and plankton were two orders of magnitude lower than requested. Integrated approaches for preparation of massive cultures of bacteria (Alcanivorax borkumensis and Marinobacter hydrocarbonoclasticus), algae (Tetraselmis suecica), rotifers (Brachionus plicatilis), and crustaceans (Artemia salina) suitable to ensure that 200 m(3) of water fulfilled the international guidelines of MEPC.174(58)G8 are here described. These methodologies allowed us to prepare the "influent water" in good agreement with guidelines and without specific problems arising from natural conditions (seasons, weather, etc.) which significantly affect the concentrations of organisms at sea. This approach also offered the chance to reliably run land tests once every two weeks.
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