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The goal of the study was to assess the impact of 2% lidocaine nebulization, a projected alternative treatment of equine asthma, on the healthy equine respiratory microbiota. A prospective, randomized, controlled, blinded, 2-way crossover study was performed, to assess the effect of 1 mg\/kg 2% lidocaine (7 treatments over 4 days) on the equine respiratory microbiota compared to control horses (saline and no treatment). Clinical assessments and respiratory samples, including nasal wash, endoscopic tracheal aspirate and bronchoalveolar lavage fluid, were obtained at each sample collection timepoint. The profile of the respiratory bacterial microbiota was evaluated using 16S amplicon sequencing, and clinical data compared using related samples analyses, based on data normality. The treatment did not affect the clinical data or alter the tracheal and nasal microbiota in healthy horses. However, time explained 12.6% of microbiota variation among samples. A significant difference in bacterial composition was observed between nasal and tracheal samples, showing the greatest relative abundance of Actinobacteria and Firmicutes, respectively. Bacterial DNA from bronchoalveolar lavage fluid did not amplify with generic primers targeting the V4 variable region of the prokaryotic small subunit ribosomal RNA gene, despite attempting multiple DNA extraction methods and PCR protocols, and after excluding PCR inhibition. This observation indicates that bronchoalveolar lavage fluid of healthy horses has a low bacterial load.<\/jats:p>","DOI":"10.1371\/journal.pone.0316079","type":"journal-article","created":{"date-parts":[[2025,1,24]],"date-time":"2025-01-24T18:32:41Z","timestamp":1737743561000},"page":"e0316079","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Nebulization of 2% lidocaine has no detectable impact on the healthy equine respiratory microbiota"],"prefix":"10.1371","volume":"20","author":[{"given":"Lauren","family":"Holley","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah N.","family":"Creasey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8451-8306","authenticated-orcid":true,"given":"Daniela","family":"Bedenice","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Reed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6413-6861","authenticated-orcid":true,"given":"Debora Regina","family":"Romualdo da Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Trautwein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa","family":"Mazan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Widmer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,1,24]]},"reference":[{"issue":"4","key":"pone.0316079.ref001","doi-asserted-by":"crossref","DOI":"10.1183\/13993003.02086-2016","article-title":"The microbiome in respiratory medicine: current challenges and future perspectives","volume":"49","author":"R Faner","year":"2017","journal-title":"European Respiratory Journal"},{"issue":"3","key":"pone.0316079.ref002","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1586\/ers.13.24","article-title":"The role of the bacterial microbiome in lung disease","volume":"7","author":"RP Dickson","year":"2013","journal-title":"Expert review of respiratory medicine"},{"key":"pone.0316079.ref003","doi-asserted-by":"crossref","DOI":"10.1155\/2016\/6470364","article-title":"Does IL-17 respond to the disordered lung microbiome and contribute to the neutrophilic phenotype in asthma?","volume":"2016","author":"X Yang","year":"2016","journal-title":"Mediators of inflammation"},{"issue":"11","key":"pone.0316079.ref004","doi-asserted-by":"crossref","first-page":"1967","DOI":"10.3390\/jcm8111967","article-title":"Lung microbiome in asthma: current perspectives","volume":"8","author":"K Loverdos","year":"2019","journal-title":"Journal of Clinical Medicine"},{"issue":"2","key":"pone.0316079.ref005","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.jaci.2010.10.048","article-title":"Airway microbiota and bronchial hyperresponsiveness in patients with suboptimally controlled asthma","volume":"127","author":"YJ Huang","year":"2011","journal-title":"Journal of Allergy and Clinical Immunology"},{"issue":"1","key":"pone.0316079.ref006","first-page":"1","article-title":"Upper and lower respiratory tract microbiota in horses: bacterial communities associated with health and mild asthma (inflammatory airway disease) and effects of dexamethasone","volume":"17","author":"SL Bond","year":"2017","journal-title":"BMC microbiology"},{"issue":"2","key":"pone.0316079.ref007","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1165\/rcmb.2017-0228OC","article-title":"Lung microbiome is influenced by the environment and asthmatic status in an equine model of asthma","volume":"60","author":"G Fillion-Bertrand","year":"2019","journal-title":"American journal of respiratory cell and molecular biology"},{"key":"pone.0316079.ref008","volume-title":"The human respiratory microbiome: implications and impact. 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