{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T12:02:46Z","timestamp":1774958566866,"version":"3.50.1"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,7]],"date-time":"2025-07-07T00:00:00Z","timestamp":1751846400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>Borrelia afzelii<\/jats:italic> and <jats:italic>Borrelia garinii<\/jats:italic> are the leading causes of Lyme borreliosis (LB) in Europe. Persistent LB forms may involve biofilms, potentially contributing to antibiotic tolerance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Whole genome sequencing (WGS) was conducted on 7 <jats:italic>B. afzelii<\/jats:italic> and 5 <jats:italic>B. garinii<\/jats:italic> isolates from erythema migrans skin biopsies. Biofilms were analyzed for extracellular DNA (eDNA) content and biomass. A phenol red metabolic assay assessed the minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) of amoxicillin, azithromycin, ceftriaxone, and doxycycline.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Phylogenetic analysis revealed <jats:italic>B. afzelii<\/jats:italic> and <jats:italic>B. garinii<\/jats:italic> formed distinct clades, while <jats:italic>B. burgdorferi<\/jats:italic> B31 clustered separately. Core genome analysis showed 38.9% of genes were shared between <jats:italic>B. afzelii<\/jats:italic> and <jats:italic>B. garinii<\/jats:italic>, decreasing to 26.1% with <jats:italic>B. burgdorferi<\/jats:italic>. The cloud genome expanded from 34.4% to 53.4% with the addition of <jats:italic>B. burgdorferi<\/jats:italic>. No antimicrobial resistance genes were detected. Surface adhesion gene profiles exhibited significant variation across species, suggesting potential functional differences in host adaptation. <jats:italic>B. afzelii<\/jats:italic> and <jats:italic>B. garinii<\/jats:italic> species exhibited biofilms, with biomass correlating significantly with eDNA production. MIC values were 0.25 \u03bcg\/mL (amoxicillin, ceftriaxone), 0.125 \u03bcg\/mL (azithromycin), and 0.5 \u03bcg\/mL (doxycycline), with no significant interspecies differences. However, MBIC values were considerably higher: 2 \u03bcg\/mL (amoxicillin, azithromycin), 16 \u03bcg\/mL (ceftriaxone), and 32 \u03bcg\/mL (doxycycline).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Biofilms in <jats:italic>B. afzelii<\/jats:italic> and <jats:italic>B. garinii<\/jats:italic> significantly reduce antibiotic efficacy, particularly ceftriaxone and doxycycline. These <jats:italic>in vitro<\/jats:italic> findings highlight the need for targeted therapeutic strategies and suggest biofilms may impact treatment outcomes in LB.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2025.1619660","type":"journal-article","created":{"date-parts":[[2025,7,7]],"date-time":"2025-07-07T05:26:57Z","timestamp":1751866017000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Genomic characterization and antibiotic susceptibility of biofilm-forming Borrelia afzelii and Borrelia garinii from patients with erythema migrans"],"prefix":"10.3389","volume":"15","author":[{"given":"Giorgia","family":"Fabrizio","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ilaria","family":"Cavallo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Sivori","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mauro","family":"Truglio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniela","family":"Kovacs","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Massimo","family":"Francalancia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanna","family":"D\u2019Agosto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisabetta","family":"Trento","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Grazia","family":"Prignano","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arianna","family":"Mastrofrancesco","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"Ruzi\u010d-Sablji\u010d","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fulvia","family":"Pimpinelli","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Enea Gino","family":"Di Domenico","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"D690","DOI":"10.1093\/nar\/gkac920","article-title":"CARD 2023: expanded curation, support for machine learning, and resistome prediction at the Comprehensive Antibiotic Resistance Database","volume":"51","author":"Alcock","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"B2","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/978-94-007-0940-9_3","article-title":"Adhesion mechanisms of Borrelia burgdorferi","volume-title":"Bacterial Adhesion","author":"Antonara","year":"2011"},{"key":"B3","doi-asserted-by":"publisher","first-page":"2652","DOI":"10.1172\/JCI58813","article-title":"Spirochete antigens persist near cartilage after murine Lyme borreliosis therapy","volume":"122","author":"Bockenstedt","year":"2012","journal-title":"J. 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