Combined effects of binary antibiotic mixture on growth, microcystin production, and extracellular release of Microcystis aeruginosa : application of response surface methodology

Abstract

The interactive effects of binary antibiotic mixtures of spiramycin (SP) and ampicillin (AMP) on Microcystis aeruginosa (MA) in terms of growth as well as microcystin production and extracellular release were investigated through the response surface methodology (RSM). SP with higher 50 and 5% effective concentrations in MA growth was more toxic to MA than AMP. RSM model for toxic unit approach suggested that the combined toxicity of SP and AMP varied from synergism to antagonism with SP/AMP mixture ratio decreasing from reversed equitoxic ratio (5:1) to equitoxic ratio (1:5). Deviations from the prediction of concentration addition (CA) and independent action (IA) model further indicated that combined toxicity of target antibiotics mixed in equivalent ratio (1:1) varied from synergism to antagonism with increasing total dose of SP and AMP. With the increase of SP/AMP mixture ratio, combined effect of mixed antibiotics on MA growth changed from stimulation to inhibition due to the variation of the combined toxicity and the increasing proportion of higher toxic component (SP) in the mixture. The mixture of target antibiotics at their environmentally relevant concentrations with increased total dose and SP/AMP mixture ratio stimulated intracellular microcystin synthesis and facilitated MA cell lysis, thus leading to the increase of microcystin productivity and extracellular release.

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