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To date, no exopolysaccharide clearly contributing to NT<jats:italic>Hi<\/jats:italic> biofilms has been identified. Consequently, there is some debate as to whether NT<jats:italic>Hi<\/jats:italic> forms biofilms during colonization and infection. The present work shows that NT<jats:italic>Hi<\/jats:italic> can form biofilms <jats:italic>in vitro,<\/jats:italic> producing an extracellular matrix composed of proteins, nucleic acids, and a \u03b2-glucan. Extracellular DNA, visualized by immunostaining and using fluorochromes, is an important component of this matrix and appears to be essential in biofilm maintenance. Extracellular RNA appears to be required only in the first steps of biofilm formation. Evidence of a matrix polysaccharide was obtained by staining with Calcofluor white M2R and by disaggregating biofilms with cellulase. Using strain 54997, residues of Glc<jats:italic>p<\/jats:italic>(1\u21924) in the NT<jats:italic>Hi<\/jats:italic> biofilm were confirmed by gas-liquid chromatography-mass spectrometry. Evidence that <jats:italic>N<\/jats:italic>-acetyl-L-cysteine shows notable killing activity towards <jats:italic>in vitro<\/jats:italic> NT<jats:italic>Hi<\/jats:italic> biofilm-forming bacteria is also provided.<\/jats:p>","DOI":"10.1038\/srep36424","type":"journal-article","created":{"date-parts":[[2016,11,2]],"date-time":"2016-11-02T10:50:47Z","timestamp":1478083847000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":47,"title":["Evidence of the presence of nucleic acids and \u03b2-glucan in the matrix of non-typeable Haemophilus influenzae in vitro biofilms"],"prefix":"10.1038","volume":"6","author":[{"given":"Mirian","family":"Domenech","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Pedrero-Vega","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicia","family":"Prieto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ernesto","family":"Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,11,2]]},"reference":[{"key":"BFsrep36424_CR1","doi-asserted-by":"publisher","first-page":"294","DOI":"10.1128\/MMBR.62.2.294-308.1998","volume":"62","author":"AR Foxwell","year":"1998","unstructured":"Foxwell, A. 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