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The presence of biofilms may play an important role in the resistance of otic pathogens to antimicrobial agents. Biofilm production of twenty <jats:italic>Staphylococcus pseudintermedius<\/jats:italic> and twenty <jats:italic>Pseudomonas aeruginosa<\/jats:italic> canine otic isolates was determined quantitatively using a microtiter plate assay, and each isolate was classified as a strong, moderate, weak or nonbiofilm producer. Minimum biofilm eradication concentration (MBEC) of two ionophores (narasin and monensin) and three adjuvants (N\u2010acetylcysteine (NAC), Tris\u2010EDTA and disodium EDTA) were investigated spectrophotometrically (OD<jats:sub>570nm<\/jats:sub>) and quantitatively (CFU\/ml) against selected <jats:italic>Staphylococcus<\/jats:italic> and <jats:italic>Pseudomonas<\/jats:italic> biofilm cultures. Concurrently, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of planktonic cultures were assessed. 16\/20 of the <jats:italic>S.\u00a0pseudintermedius<\/jats:italic> clinical isolates were weak biofilm producers. 19\/20 <jats:italic>P.\u00a0aeruginosa<\/jats:italic> clinical isolates produced biofilms and were distributed almost equally as weak, moderate and strong biofilm producers. While significant antibiofilm activity was observed, no MBEC was achieved with narasin or monensin. The MBEC for NAC ranged from 5,000\u201310,000\u00a0\u00b5g\/ml and from 20,000\u201380,000\u00a0\u00b5g\/ml against <jats:italic>S.\u00a0pseudintermedius<\/jats:italic> and <jats:italic>P.\u00a0aeruginosa<\/jats:italic>, respectively. Tris\u2010EDTA eradicated <jats:italic>P.\u00a0aeruginosa<\/jats:italic> biofilms at concentrations ranging from 6,000\/1,900 to 12,000\/3,800\u00a0\u00b5g\/ml. The MBEC was up to 16\u2010fold and eightfold higher than the MIC\/MBC of NAC and Tris\u2010EDTA, respectively. Disodium EDTA reduced biofilm growth of both strains at concentrations of 470\u00a0\u00b5g\/ml and higher. It can be concluded that biofilm production is common in pathogens associated with canine OE. NAC and Tris\u2010EDTA are effective antibiofilm agents <jats:italic>in vitro<\/jats:italic> that could be considered for the treatment of biofilm\u2010associated OE in dogs.<\/jats:p>","DOI":"10.1111\/jvp.12811","type":"journal-article","created":{"date-parts":[[2019,9,10]],"date-time":"2019-09-10T13:15:22Z","timestamp":1568121322000},"page":"682-692","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":37,"title":["Biofilm production by pathogens associated with canine otitis externa, and the antibiofilm activity of ionophores and antimicrobial adjuvants"],"prefix":"10.1111","volume":"42","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9415-9893","authenticated-orcid":false,"given":"Wei Yee","family":"Chan","sequence":"first","affiliation":[{"name":"Australian Centre for Antimicrobial Resistance Ecology School of Animal and Veterinary Sciences The University of Adelaide Roseworthy SA Australia"},{"name":"Faculty of Veterinary Medicine Universiti Putra Malaysia Selangor Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4862-503X","authenticated-orcid":false,"given":"Elizabeth E.","family":"Hickey","sequence":"additional","affiliation":[{"name":"Australian Centre for Antimicrobial Resistance Ecology School of Animal and Veterinary Sciences The University of Adelaide Roseworthy SA Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5312-2149","authenticated-orcid":false,"given":"Stephen W.","family":"Page","sequence":"additional","affiliation":[{"name":"Luoda Pharma Pty. 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