Abstract
A large number of antibiotics are entering the aquatic environment accompanying human and animal excreta, which will threaten the survival of aquatic organisms and even human health. It has been found that binary mixtures of aminoglycoside (AG) exhibit additive action and can be evaluated well by a classical model, concentration addition (CA) in our past study. Therefore, to investigate the toxicity interaction within multi-component mixtures of AG antibiotics, five antibiotics, kanamycin sulfate (KAN), neomycin sulfate (NEO), tobramycin (TOB), streptomycin sulfate (STS), and gentamicin sulfate (GEN), were selected to construct five-component mixture systems by a uniform design ray method. The toxic effects (luminescence inhibition) of single antibiotic and five-antibiotic mixture systems towards a photobacterium Vibrio qinghaiensis sp.-Q67 (V. qinghaiensis) in different exposure time (0.25, 2, 4, 8, and 12 h) were determined by the time-dependent microplate toxicity analysis method. The concentration-effect data were fitted by a nonlinear least square method, toxicity interaction within mixture systems was analyzed by a CA model, and the interaction intensity was characterized by deviation from the CA model (dCA). Besides, the toxicity mechanism of five antibiotics and their five-component mixtures to V. qinghaiensis was analyzed by electron microscopy. The results show that toxicity of five antibiotics and their five-component mixture systems to V. qinghaiensis is time-dependent and has strong long-term toxicity. Different from binary AG antibiotic mixture systems, five-antibiotic mixture systems exhibit obviously time-dependent synergism. In addition, toxicity of the five-antibiotic mixtures can be 1.4 times higher than that of the mixtures without synergisms at the same concentration level. According to dCA, synergism intensity (dCA) curves of rays move slowly from the high concentration region to the medium or lower one and the maximum dCA values also increase, decrease, or first increase, then decrease with the lengthening of exposure time. The inhibition activity and synergism intensity of mixture rays have good correlation with the concentration ratios of STS, the key component for synergism. The cell morphology of V. qinghaiensis indicates the strong toxicity of five antibiotics and their mixture rays is not due to the destruction of cell structure, but the inhibition of the light-emitting activity of the photobacterium.