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Extracellular DNA plays a crucial role in biofilm formation and self-defence, with nucleases being proposed as promising agents for biofilm disruption. This study evaluated the in vitro and in vivo efficacy of DNase I in improving the activity of cefotaxime, amikacin, and ciprofloxacin against K. pneumoniae biofilms. K. pneumoniae ATCC 700603 and a clinical isolate from catheter-related bloodstream infection were cultured for biofilm formation on microtiter plates, and the antibiofilm activity of the antibiotics (0.03\u201364\u2009mg\/L), with or without bovine pancreatic DNase I (1\u201332\u2009mg\/L) was determined by XTT dye reduction test and viable counting. The effect of ciprofloxacin (2\u2009mg\/L) and DNase I (16\u2009mg\/L) was further evaluated in vitro on 1-cm-long silicon catheter segments, and in a mouse model of subcutaneous catheter-associated infection. Combination with DNase I did not improve the biofilm-preventive capacity of the three antibiotics or the biofilm-eradicating capacity of cefotaxime and amikacin. The biofilm-eradicating capacity of ciprofloxacin was increased by 8-fold and 4-fold in K. pneumoniae ATCC 700603 and clinical isolate, respectively, with DNase I. The combination therapy caused 99% reduction in biofilm biomass in the mouse model.<\/jats:p>","DOI":"10.1093\/femspd\/ftad001","type":"journal-article","created":{"date-parts":[[2023,1,12]],"date-time":"2023-01-12T13:36:39Z","timestamp":1673530599000},"source":"Crossref","is-referenced-by-count":14,"title":["<i>In vitro<\/i>\n                    and\n                    <i>in vivo<\/i>\n                    evaluation of DNase I in reinstating antibiotic efficacy against\n                    <i>Klebsiella pneumoniae<\/i>\n                    biofilms"],"prefix":"10.1093","volume":"81","author":[{"given":"Anayata","family":"Sharma","sequence":"first","affiliation":[{"name":"Department of Microbial Biotechnology, Panjab University , Chandigarh, 160014 , India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5425-8064","authenticated-orcid":false,"given":"Praveen","family":"Rishi","sequence":"additional","affiliation":[{"name":"Department of 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