Evaluation of residual toxicity of hypochlorite-treated water using bioluminescent microbes and microalgae: Implications for ballast water management.

Total residual oxidants (TRO) in treated ballast water can produce various disinfection by-products (DBPs) depending on local conditions, such as salinity and organic matter content in water. Because TRO and DBPs are known to be harmful to aquatic organisms and humans, ecotoxicity tests have been proposed for screening the residual toxicity before discharging treated ballast water. In the present study, we aimed to address the decay rates and toxicity changes of TRO under various conditions in salinity, initial TRO concentrations, and residence time of TRO. In addition, the toxicological sensitivities of bioluminescent bacteria Vibrio fischeri and a commonly-used microalgae Skeletonema costatum relative to the residual toxicity of TRO and six selected DBPs were determined. Decay rate of TRO concentration increased as a function of salinity and was affected by the initial concentrations of TRO. Unexpectedly, significant bioluminescence inhibition was observed for hypochlorite-treated water at < 0.1 mg L-1 TRO (expressed as Cl2), which is a lower concentration than the maximum allowable discharge concentration (MADC) to marine waters established by the International Maritime Organization (IMO). The ecotoxicological thresholds of no observed effective concentration and median effect concentration for all tested DBPs were about 3-10 times lower for V. fischeri than for S. costatum. The results indicate that bioluminescent microbes possess an ecologically-relevant sensitivity to both TRO and DBPs in ballast water. In general, bioassay using V. fischeri was potentially more effective than microalgae for screening the total toxicity of TRO and DBPs in treated ballast water, especially given that ballast water usually contains a highly variable and complex mixture of toxicants.

Validation of algal viability treated with total residual oxidant and organic matter by flow cytometry.

Algal cell growth after starch and oxidant treatments in seawater species (Isochrysis galbana and Phaeodactylum tricornutum) and freshwater species (Selenastrum capricornutum and Scenedesmus obliquus) were evaluated by flow cytometry with fluorescein diacetate (FDA) staining to determine algal viability. Growth of algal cell was found to be significantly different among groups treated with NaOCl, starch and/or sodium thiosulfate, which are active substance (Total Residual Oxidant; TRO as Cl2), organic compound to meet efficacy testing standard and neutralizer of TRO by Ballast Water Management Convention of International Maritime Organization, respectively. The viability of algal cell treated with TRO in starch-add culture of 5days after treatment and neutralization was decreased significantly. ATP contents of the treated algal cells corresponded to the FL1 fluorescent signal of flow cytometry with FDA staining. I. galbana was the most sensitive to TRO-neutralized cultures during viability analysis.