Acute toxicity of arsenic to Aliivibrio fischeri (Microtox® bioassay) as influenced by potential competitive–protective agents

In this study, we investigated the effect of some potential alleviative compounds against the acute toxicity of arsenic (As^V, As^III and DMA^V) on Aliivibrio fischeri (formerly Vibrio fischeri), a bioluminescent model bacterium, through the Microtox® bioassay. The compounds studied differed in their mechanism of action, and they included the following: phosphate and glycerol, as chemical analogues (and potential competitors) of As^V or As^III, respectively; citrate, a weak natural organic ligand; and the antioxidant ascorbic acid. Special attention was paid to phosphate effects, a widespread pollutant in natural environments. As^V was found to be more acutely toxic than As^III to A. fischeri, in accordance with its higher interaction with the bacteria. Both As^V and As^III were found to be much more acutely toxic than DMA^V, which was essentially non-acutely toxic even at very high concentrations. Phosphate presence (at equimolar P/As ratios or higher) resulted in the almost total suppression of bioluminescence inhibition, suggesting it exerts an alleviative effect against As^V acute toxicity on A. fischeri. Interestingly, the uptake and the percentage of extracellular As^V were not affected by the addition of phosphate, suggesting that such protective effect does not result from the competition for their common transporters. In contrast, the acute toxicity of As^III was essentially unaffected by phosphate. Glycerol did not decrease the acute toxicity or the uptake of As^III by A. fischeri, denoting the likely occurrence of an additional mechanism for As^III uptake in such bacteria. Similarly, citrate and ascorbic acid essentially did not caused alleviation of As^V or As^III acute toxicity. As for environmental and operational implications, P could beneficially protect aquatic microorganisms against acute detrimental effects of As^V, whilst its presence could mask the toxicity due to As^V when assessed using the Microtox® bioassay, thus leading to seriously underestimate the actual ecological and health risks.