Toxicity of commercial and pure forms of three nonsteroidal anti-inflammatory drugs in Xenopus laevis embryos before and after ozonation.

In this study, the toxic and teratogenic effects of three commercial drugs and their active ingredients on Xenopus laevis embryos before and after ozonation were evaluated using the Frog Embryos Teratogenesis Assay-Xenopus (FETAX). First, the median lethal concentration (LC50) and, if data were available, the median effective concentration, teratogenic index and minimum growth inhibitory concentration were determined for each drug substance without ozonation. Then, the active substance amounts of three selected nominal concentrations (LC50/2, LC50, and LC50×2) of each test substance before ozonation were measured by HPLC analysis and the toxicity of these substances was evaluated after 2, 3, 4, and 5 h of ozonation. In addition, degradation products that may occur during ozonation were evaluated by LC-MS analysis. The 96-h LC50s of Dolphin-diflunisal, Dichloron-diclofenac sodium, and Apranax-naproxen drug-active substance pairs were determined to be 22.3 and 11.1, 25.7 and 18.7, and 47.8 mg active substance/L and 45.3 mg/L, respectively. According to the FETAX test results, the Dolphin-diflunisal drug-active ingredient pair did not cause growth retardation in exposed embryos. Dichloron-diclofenac sodium and Apranax-naproxen drug-active ingredient pairs were both teratogenic and growth inhibitory. In the second stage of the study, in which the effectiveness of ozonation in eliminating the toxic effects of drugs is evaluated, it is seen that ozonation is partially successful in eliminating the toxic effects of Dolphin-diflunisal and Dichloron-diclofenac sodium pairs, but insufficient for eliminating the effects of the Apranax-naproxen pair.

Biochemical Studies to Understand Teratogenicity and Lethality Outcomes in Modified-FETAX.

The frog embryo teratogenesis assay-Xenopus (FETAX) is a standardized test used to assess the toxic and teratogenic effects of xenobiotics. With this test, toxic and/or teratogenic concentrations of xenobiotic substances can be determined using morphological parameters such as lethality, length, and malformations in stage 8-11 Xenopus laevis embryos after 96 h exposure. These parameters enable the determination of the median lethal and effective concentrations (LC50 and EC50), minimum concentration to inhibit growth (MCIG), and teratogenic index of the tested chemical to reveal the short-term effects of relatively high concentrations. On the other hand, although FETAX provides quantitative and qualitative data on teratogenicity and toxicity, the biochemical and molecular mechanisms of these effects cannot be explained. Recent studies have tried to elucidate the mechanisms causing malformations and to explain the underlying causes of toxicity and teratogenicity by biochemical marker analysis. This chapter describes methods to analyze modified-FETAX and some detoxification and oxidative stress-related biomarkers during the early embryonic development of X. laevis.

Assessing the impact of antiviral drugs commonly utilized during the COVID-19 pandemic on the embryonic development of Xenopus laevis.

The antiviral drugs favipiravir and oseltamivir are widely used to treat viral infections, including coronavirus 2019 (COVID-19), and their levels are expected to increase in the aquatic environment. In this study, the potential toxic and teratogenic effects of these drugs were evaluated using the frog embryo teratogenesis assay Xenopus (FETAX). In addition, glutathione S-transferase (GST), glutathione reductase (GR), catalase, carboxylesterase (CaE), and acetylcholinesterase (AChE) enzyme activities and malondialdehyde levels were measured as biochemical markers in embryos and tadpoles for comparative assessment of the sublethal effects of the test compounds. Prior to embryo exposure, drug concentrations in the exposure medium were measured with high-performance liquid chromatography. The 96-h median lethal concentration (LC50) was 137.9 and 32.3 mg/L for favipiravir and oseltamivir, respectively. The teratogenic index for favipiravir was 4.67. Both favipiravir and oseltamivir inhibited GR, CaE, and AChE activities in embryos, while favipiravir increased the GST and CaE activities in tadpoles. In conclusion, favipiravir, for which teratogenicity data are available in mammalian test organisms and human teratogenicity is controversial, inhibited Xenopus laevis embryo development and was teratogenic. In addition, sublethal concentrations of both drugs altered the biochemical responses in embryos and tadpoles, with differences between the developmental stages.