Carvalho, Sandra M.Freitas, Cláudia S.Oliveira, Ana S.Saraiva, Lígia M.2022-07-202022-07-202021-09-010378-1097PURE: 36371588PURE UUID: 7ab49073-4e59-40d0-bbf3-14c21948026aScopus: 85119322427PubMed: 34665259http://hdl.handle.net/10362/142212Publisher Copyright: © 2021 The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.Staphylococcus aureus is a Gram-positive bacterium with capacity to form biofilms, which constitute an important resistance mechanism and virulence factor. Flavohaemoglobin (Hmp) is a major nitric oxide (NO) detoxifier of several bacteria, including S. aureus. Although Hmp has a well-known physiological role linked to response of planktonic cells to nitrosative stress, its contribution to biofilm formation remains unaddressed. Hence, in this work, we investigated the role of Hmp in biofilm development of a methicillin-resistant S. aureus strain. For this purpose, we exposed the hmp mutant to nitrosative stress and examined its behaviour along biofilm development. We observed that cells inactivated in hmp and grown under nitrosative stress conditions have significantly impaired capacity to develop early stage biofilms. Furthermore, the wild-type biofilm phenotype was fully restored by trans-complementation of hmp in the hmp mutant. Coculture studies of NO-producing macrophages with S. aureus revealed that the hmp mutant has significantly lower capacity to develop biofilm biomass when compared with the wild type. Thus, we concluded that the pathogen S. aureus relies on Hmp to establish viable biofilms in the presence of cells of the host innate immune system.1216641engearly stage biofilmflavohaemoglobinmacrophagesmethicillin-resistant Staphylococcus aureusnitric oxidenitrosative stressMicrobiologyMolecular BiologyGeneticsjournal article10.1093/femsle/fnab131https://www.scopus.com/pages/publications/85119322427