Bordetella pertussis is the etiological agent of whooping cough, a common cause of respiratory illness in both children and adults. In the present study, we investigated the bactericidal activity of four antiseptics-povidone-iodine (PVP-I), benzethonium chloride (BEC), chlorhexidine gluconate (CHG) and benzalkonium chloride (BAC)-against B. pertussis ATCC9797 and clinical isolates. Among the topical antiseptics, PVP-I, BEC, and BAC, PVP-I and BAC in particular, showed high bactericidal activity, whereas CHG had low activity. PVP-I gargle also showed high bactericidal activity, similar to topical PVP-I. However, BEC gargle had low bactericidal activity. Our results indicate that topical PVP-I and BAC, and PVP-I gargle would be useful as effective antiseptics against B. pertussis.