The use of animals in education and research is a controversial issue that involves ethical considerations. In Brazil, Act 11,794, which was approved in 2008, established the National Council on the Control of Animal Experimentation (CONCEA) and a database of institutions that use animals for research and education (CIUCA). This legislation also set out the regulations for the use of animals. In this study, we have evaluated the ethical issues involved in the use of animals for educational purposes at the Federal University of Paraná, through a qualitative-quantitative analysis that relied on written questionnaires. Our objective was to find out the opinions of students and staff from different academic fields, and at different stages in their professional development, on the use of animals for educational purposes. The study involved 101 students and 20 lecturers (i.e. tenure-track professors and all those who teach the students) in Biology, Pharmacology, Medicine and Veterinary Medicine. Approximately half of the students (45.5%) did not know the legislation that regulates the use of animals in education, and most of the lecturers believed that learning goals could not be achieved with alternative methods. Only 38.9% of the lecturers and 31.9% of the students trusted the usefulness of alternative methods. Furthermore, recent graduates were as unaware of the legislation, as were students in the first two years of their university courses. These results suggest that it is necessary to considerably expand the discussion on alternatives to animal use in the academic environment.