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Penicillin, metformin, and aluminum are commonly used substances and their presence in the environment has increased due to their widespread use. In water bodies, the combined presence of xenobiotics leads to additive, synergistic, or antagonistic interactions that significantly modify the toxic response, being more evident in the early stages of development of individuals. These interactions can be evaluated through biomarkers such as the activity and expression of antioxidant enzymes and the production of congenital malformations in exposed organisms. However, little is known about the effects that mixtures of drugs such as penicillin, metformin, and metals such as aluminum can produce on aquatic species that are constantly exposed to these xenobiotics such as common carp. The objective of this study was to determine the toxicity and type of interaction produced on Cyprinus carpio embryos exposed to these pollutants isolated and in mixtures. Here we show that the mixture of these pollutants produces antagonism and synergism at high and low concentrations respectively. In addition, toxicity results show that in embryos subacutely exposed (NOAEL) to the mixture of contaminants, the activity of antioxidant enzymes and their gene expression (PCR-RT) is increased, embryonic development is modified, and teratogenesis occurs. As can be observed, the mixture of the contaminants influenced the toxic response, evidencing the importance of continuing to study interactions; since this is the way, they are usually found in contaminated bodies of water. Highlights: 1. Exposure of Cyprinus carpio embryos to the mixture of metformin, penicillin and aluminum at high concentrations (LC50) produces an antagonistic type of interaction. 2. Exposure of Cyprinus carpio embryos to the mixture of metformin, penicillin and aluminum at sublethal concentrations (NOAEL) produces a synergistic type interaction. 3. Antioxidant enzyme activity of Cyprinus carpio embryos exposed to NOAEL-equivalent concentrations of metformin, penicillin and aluminum is modified by simultaneous exposure to the toxicants. 4. Gene expression of antioxidant enzymes in Cyprinus carpio embryos exposed to NOAEL-equivalent concentrations of metformin, penicillin and aluminum is increased by simultaneous exposure to the toxicants during the first hours of development. 5. The embryonic developmental score of Cyprinus carpio is decreased by simultaneous exposure to metformin, penicillin and aluminum. [ABSTRACT FROM AUTHOR]

Aluminum, mercury, and iron have been found in high concentrations in various freshwater bodies around the world and have been shown to be very harmful contaminants to hydrobionts, causing metabolic dysfunction and damage at various levels. However, studies on mixtures of these pollutants are scarce, particularly in younger or developing organisms, which are more sensitive to damage. Therefore, the objective of this work was to evaluate the toxicity of Al, Fe, and Hg, in isolation and in mixture, on common carp embryos exposed to the maximum permissible limits described in the Mexican regulations for the protection of aquatic life, correlating the biomarkers of oxidative stress with the effects on embryonic development. For this purpose, the Cyprinus carpio embryos were exposed to iron (0.1 mg L−1), mercury (0.00001 mg L−1), aluminum (0.05 mg L−1), and their mixture, throughout their development and until hatching. The activity of the antioxidant enzymes (SOD, CAT, and GPx), the degree of lipoperoxidation, and the content of hydroperoxides and total proteins, as well as the morphological development of the embryos were evaluated at 12 h, 24 h, 48 h, 72 h, and 96 h of exposure. The results showed that the metals under study are toxic to C. carpio embryos and that their interaction modifies the toxic response. Thus, iron generates alterations in the activity of antioxidant enzymes and modification in embryonic development to a lesser extent than aluminum and the mixture of metals, while mercury exerts its toxicity on embryos by mechanisms other than oxidative stress, but the modification to the activity of antioxidant enzymes may contribute to the changes observed in embryonic development. The changes in the response of biomarkers of embryotoxicity and oxidative stress suggest that the combination of metals produces an antagonism-type interaction, so this study provides a precedent for future research to determine the type of interaction that a mixture of contaminants generates in developing aquatic organisms. [ABSTRACT FROM AUTHOR]