{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T01:15:47Z","timestamp":1788570947309,"version":"build-2803163510"},"reference-count":62,"publisher":"Oxford University Press (OUP)","issue":"7","license":[{"start":{"date-parts":[[2023,5,30]],"date-time":"2023-05-30T00:00:00Z","timestamp":1685404800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"name":"\u00d6rebro County Council Research Committee","award":["AI47609"],"award-info":[{"award-number":["AI47609"]}]},{"DOI":"10.13039\/501100002915","name":"Foundation for Medical Research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002915","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000738","name":"Department of Veterans Affairs","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000738","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["AI47609"],"award-info":[{"award-number":["AI47609"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,7,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Objectives<\/jats:title>\n                  <jats:p>Gentamicin is used in several alternative treatments for gonorrhoea. Verified clinical Neisseria gonorrhoeae isolates with gentamicin resistance are mainly lacking and understanding the mechanisms for gonococcal gentamicin resistance is imperative. We selected gentamicin resistance in gonococci in vitro, identified the novel gentamicin-resistance mutations, and examined the biofitness of a high-level gentamicin-resistant mutant.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Low- and high-level gentamicin resistance was selected in WHO X (gentamicin MIC\u200a=\u200a4 mg\/L) on gentamicin-gradient agar plates. Selected mutants were whole-genome sequenced. Potential gentamicin-resistance fusA mutations were transformed into WT strains to verify their impact on gentamicin MICs. The biofitness of high-level gentamicin-resistant mutants was examined using a competitive assay in a hollow-fibre infection model.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>WHO X mutants with gentamicin MICs of up to 128 mg\/L were selected. Primarily selected fusA mutations were further investigated, and fusAR635L and fusAM520I\u200a+\u200aR635L were particularly interesting. Different mutations in fusA and ubiM were found in low-level gentamicin-resistant mutants, while fusAM520I was associated with high-level gentamicin resistance. Protein structure predictions showed that fusAM520I is located in domain IV of the elongation factor-G (EF-G). The high-level gentamicin-resistant WHO X mutant was outcompeted by the gentamicin-susceptible WHO X parental strain, suggesting lower biofitness.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>We describe the first high-level gentamicin-resistant gonococcal isolate (MIC\u200a=\u200a128 mg\/L), which was selected in vitro through experimental evolution. The most substantial increases of the gentamicin MICs were caused by mutations in fusA (G1560A and G1904T encoding EF-G M520I and R635L, respectively) and ubiM (D186N). The high-level gentamicin-resistant N. gonorrhoeae mutant showed impaired biofitness.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jac\/dkad168","type":"journal-article","created":{"date-parts":[[2023,5,30]],"date-time":"2023-05-30T17:39:24Z","timestamp":1685468364000},"page":"1769-1778","source":"Crossref","is-referenced-by-count":11,"title":["High-level <i>in vitro<\/i> resistance to gentamicin acquired in a stepwise manner in <i>Neisseria gonorrhoeae<\/i>"],"prefix":"10.1093","volume":"78","author":[{"given":"Daniel","family":"Golparian","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Microbiology, Faculty of Medicine and Health, WHO Collaborating Centre for Gonorrhoea and Other Sexually Transmitted Infections, \u00d6rebro University , \u00d6rebro , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanne","family":"Jacobsson","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Microbiology, Faculty of Medicine and Health, WHO Collaborating Centre for Gonorrhoea and Other Sexually Transmitted Infections, \u00d6rebro University , \u00d6rebro , Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Concerta L","family":"Holley","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Emory University School of Medicine , Atlanta, GA , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William M","family":"Shafer","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Emory University School of Medicine , Atlanta, GA , USA"},{"name":"The Emory Antibiotic Resistance Center, Emory University School of Medicine , Atlanta, GA , USA"},{"name":"Laboratories of Bacterial Pathogenesis, Veterans Affairs Medical Center , Decatur, GA , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1710-2081","authenticated-orcid":false,"given":"Magnus","family":"Unemo","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Microbiology, Faculty of Medicine and Health, WHO Collaborating Centre for Gonorrhoea and Other Sexually Transmitted Infections, \u00d6rebro University , \u00d6rebro , Sweden"},{"name":"Institute for Global Health, University College London (UCL) , London , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,5,30]]},"reference":[{"key":"2024053000062415700_dkad168-B1","author":"WHO","year":"2021"},{"key":"2024053000062415700_dkad168-B2","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1128\/CMR.00010-14","article-title":"Antimicrobial resistance in Neisseria gonorrhoeae in the 21st century: past, evolution, and 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