Development of an ATP assay for rapid onboard testing to detect living microorganisms in ballast water

Ballast water is a principal pathway for the introduction of pathogens and non-indigenous species to ports worldwide. The International Maritime Organization (IMO) and the United States Coast Guard (USCG) have adopted ballast water management regulations that require, e.g., the installation of shipboard ballast water management systems (BWMS). Rapid and simple analytical methods are needed to monitor whether ballast water disinfection ensures compliance with the discharge standards. In this study laboratory and full scale land-based testing was used to investigate the suitability of an adenosine triphosphate (ATP) assay for quantifying living organisms (≥ 10 and < 50 μm minimum dimension) in ballast water. In laboratory experiments the ATP assay was highly sensitive, with a detection limit of < 5 cells 0.1 mL− 1. Diatom species (Chaetoceros simplex and Skeletonema costatum) had low ATP concentrations compared with dinoflagellate, Raphidophyceae, and Chrysophyceae species. This was because of differences in cell volume, as the ATP concentration increased exponentially with increasing cell volume. Using a regression model between ATP concentration and cell volume, an estimated the pass and fail ATP concentration in this study (788-98,610 pg mL− 1) was developed for the discharge of ballast water. In land-based testing the ATP assay also showed a good correlation with the presence of living natural plankton cells in control samples, but the ATP concentration (137 pg mL− 1) was much lower than the ATP guideline. The low ATP concentration in natural plankton cells may reflect a decline in their biological activity because of extended exposure to dark conditions. Although our results need further validation, the ATP assay is a suitable tool for monitoring compliance of ballast water treatment.