Effects of ibuprofen, diclofenac and their mixture on the seminal quality parameters of Astyanax lacustris (Characiformes: Characidae)

Abstract

The ibuprofen (IBU) and diclofenac (DFC) are anti-inflammatories broadly used andfrequently found in the water. These drugs, considered Contaminants of EmergingConcern (CECs), individually or combined, can affect the endocrine system of teleosts,altering the reproductive process. The aim of this study is to analyze the seminal qualityparameters of Astyanax lacustris, using in vitro and in vivo approaches. In the in vitroassay, semen samples from each fish (n = 48) will be collected and exposed to differentconcentrations of drugs: 1) clean water (CTL, 0 ng L -1 ); 2) Ibuprofen: (IBU, 0.1; 1; 10;100; and 1000 ng L -1 ); 3) Diclofenac (DFC, 0.1; 1; 10; 100; and 1000 ng L -1 ); and 4)mixture (IBU+DFC, summing individual concentrations) for 50s, 10 minutes, and 30minutes. In the in vivo assay, sexually mature male of A. lacustris (n=80) will be dividedinto the following groups, using environmentally relevant drug concentrations: 1) cleanwater (CTL, 0 µg L -1 ); 2) Ibuprofen: (IBU, 16 µg L -1 ); 3) Diclofenac (DFC, 14 µg L -1 ); and4) mixture (IBU+DFC, 16 + 14 µg L -1 ) for 7 days. The following biomarkers will be used;1) Sperm quality: pH and seminal osmolarity, sperm morphology, kinetics, concentration,and mitochondrial activity; 2) Genotoxicity: cometa assay; 3) Fatty acid profile andlipoperoxidation in semen and testicles. Considering that gamete fertilization occurs in theaquatic environment, the effects of IBU and DFC on semen and testicular quality, willprovide insights into the effects of these anti-inflammatories on the reproductive successof teleosts.