It has been stated by researchers that the antibiotic polymyxin B sulfate (POL) is a key component inducing time-dependent antagonism in freshwater luminescent bacteria, Vibrio qinghaiensis sp.-Q67, exposed in the ternary mixture system of the ionic liquids, pesticide and antibiotics. However, the previous statement is limited to ternary and quaternary mixtures without considering situations such as the binary system. In order to prove the direct inducing of antagonism by POL in a more complete and systematic way, two categories of experiments (adding POL in non-antagonistic ternary system and decomposing antagonistic ternary system with POL into the binary system) have been conducted in this paper. The results showed that quaternary mixture systems (adding POL to non-antagonism ternary mixture, up verification) exhibit antagonistic action in a majority of rays, at some point in the experiment. However, by decomposing the antagonistic ternary mixtures with POL into binary mixtures (down verification), the combined toxicities of binary mixtures at all time points in the experiment are additive. Obviously, the POL has a significant contribution to antagonism only in the ternary and quaternary mixtures, but not in the binary mixtures. We can draw a new conclusion that the antagonism of the multi-component mixtures is induced by at least three components (including POL), with complex chemical interactions. Therefore, considering POL's influence of antagonism as an example, future environmental protection practitioners and academic researchers should construct more scenarios of mixtures when assessing the influences and reactions of certain chemicals causing pollutions.

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