(1) Background: Evidence-based practice (EBP) informs daily clinical interventions with the purpose of seeking changes to traditional practice through scientific evidence that justifies the reasons for our actions. The objectives were to describe the barriers, beliefs, and attitudes in the application of EBP among university health professionals (not doctors) and to evaluate the influence of the COVID-19 pandemic among them. (2) Methods: This prospective study is both descriptive and observational. The individuals under study were university health professionals (not doctors) from various autonomous regions within Spain, in both public and private spheres. Sociodemographic and labor-related variables linked to the research and its completion were studied. Likewise, the survey instrument Health Sciences Evidence-Based Practice questionnaire (HS-EBP) was administered to evaluate the barriers to, beliefs in, and attitudes towards evidence-based practice. (3) Results: A total of 716 responses were gathered, of which 387 were collected during the period of confinement, and 343 in the COVID-19 post-confinement period. Possible associations that might help respond to the objectives were explored through a correlational study between the sociodemographic variables and each sub-scale of the HS-EBP 30 questionnaire (n = 716). (4) Conclusions: Barriers to, beliefs in, and attitudes towards evidence-based practice are described. There is a leadership gap where line management provides insufficient motivation to follow work routines. The COVID-19 pandemic has caused immense stress among health professionals. The post-confinement group showed a significant change in the variables “beliefs and attitudes”, and likewise in the “evaluation” block, justified by the need to update knowledge and to apply evidence.