Background The variation in infection risk among individuals is thought to be partially influenced by occupational factors. This study aims to investigate the seropositivity rates of 17 common infectious agents in male airline pilots (APs), a professional group known to experience a high prevalence of cardiovascular and gastrointestinal diseases. Methodology In our study, we employed a case-control design with 100 male APs as cases, matched by age, sex, and tenure (i.e., at least five years of service) to 100 male office workers (OWs) who served as controls. We measured the IgG antibody levels to 17 pathogens using specific enzyme-linked immunosorbent assays, including herpes simplex virus 1, herpes simplex virus 2, varicella-zoster virus, Epstein-Barr virus, cytomegalovirus, human herpesvirus 6, human herpesvirus 7, Kaposi's sarcoma-associated herpesvirus, , human T-lymphotropic virus 1, BK virus, John Cunningham virus, Merkel cell polyomavirus, human papillomavirus 16, human papillomavirus 18, , and . The determination of seropositivity cutoffs for each pathogen was made in accordance with the guidelines provided by the respective kit manufacturers. Results The seropositivity rates for the 17 pathogens ranged from 1% for human T-lymphotropic virus 1 to 94% for varicella-zoster virus and were similar in both professions, except for herpes simplex virus 1 and , which were more prevalent in APs. Conclusions Our findings suggest a higher prevalence of previous infections with herpes simplex virus 1 and in APs compared to OWs. These infections may be associated with the prevalence of specific non-communicable diseases in this professional group. However, additional longitudinal studies are needed to substantiate this hypothesis.