TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae

BackgroundStreptococcus dysgalactiae (SD) is a potent pathogen associated with infections in a broad range of host species. Notably, a substantial proportion of SD isolates exhibit reduced susceptibility to tetracycline but lack identifiable resistance determinants. In the present study, we wanted t...

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Main Authors: Marte Glambek, Morten Kjos, Marita T. Mårli, Zhian Salehian, Steinar Skrede, Audun Sivertsen, Bård R. Kittang, Oddvar Oppegaard
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Cellular and Infection Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2025.1583926/full
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author Marte Glambek
Marte Glambek
Morten Kjos
Marita T. Mårli
Zhian Salehian
Steinar Skrede
Steinar Skrede
Audun Sivertsen
Bård R. Kittang
Bård R. Kittang
Oddvar Oppegaard
Oddvar Oppegaard
author_facet Marte Glambek
Marte Glambek
Morten Kjos
Marita T. Mårli
Zhian Salehian
Steinar Skrede
Steinar Skrede
Audun Sivertsen
Bård R. Kittang
Bård R. Kittang
Oddvar Oppegaard
Oddvar Oppegaard
author_sort Marte Glambek
collection DOAJ
description BackgroundStreptococcus dysgalactiae (SD) is a potent pathogen associated with infections in a broad range of host species. Notably, a substantial proportion of SD isolates exhibit reduced susceptibility to tetracycline but lack identifiable resistance determinants. In the present study, we wanted to explore the genetic basis for this low-grade resistance to tetracycline.MethodsGenome-wide association studies were performed on a collection of 407 SD genomes to identify potential novel resistance determinants. Two strains of SD, belonging to each of the subspecies dysgalactiae and equisimilis were used for mutagenesis. Natural transformation was exploited to knock out resistance gene candidates, and the resultant mutants were compared with their respective wildtypes regarding susceptibility to tetracycline, doxycycline, minocycline, tigecycline, erythromycin, gentamicin, clindamycin and ciprofloxacin.ResultsWe identified a two gene operon, herein designated trexAB, significantly associated with reduced susceptibility to tetracycline. The proteins encoded by the operon were predicted in silico to constitute a heterodimeric efflux transporter. The knockout of trexAB led to a 16- to 32-fold reduction in minimum inhibitory concentration (MIC) for tetracycline and a 4-fold reduction in MIC for tigecycline in the investigated strains. No differences between mutants and wildtypes were observed for other antibiotics included in the test panel. Whole genome alignment of mutants and their respective wildtypes revealed no differences other than the expected differences caused by the knockout.ConclusionWe have characterized a novel operon causing low-grade resistance to tetracycline in SD. The MIC distribution of trexAB-positive isolates is intersected by the current EUCAST susceptibility breakpoint, and our findings are relevant for future revisions and determinations of adequate breakpoints for tetracycline in S. dysgalactiae.
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spelling doaj-art-bebd54af752a4ed5813f59b7960a48bd2025-08-20T03:27:36ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-06-011510.3389/fcimb.2025.15839261583926TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiaeMarte Glambek0Marte Glambek1Morten Kjos2Marita T. Mårli3Zhian Salehian4Steinar Skrede5Steinar Skrede6Audun Sivertsen7Bård R. Kittang8Bård R. Kittang9Oddvar Oppegaard10Oddvar Oppegaard11Department of Medicine, Haukeland University Hospital, Bergen, NorwayDepartment of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, NorwayFaculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, NorwayFaculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, NorwayFaculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, NorwayDepartment of Medicine, Haukeland University Hospital, Bergen, NorwayDepartment of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, NorwayDepartment of Microbiology, Haukeland University Hospital, Bergen, NorwayDepartment of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, NorwayDepartment of Internal Medicine, Haraldsplass Deaconess Hospital, Bergen, NorwayDepartment of Medicine, Haukeland University Hospital, Bergen, NorwayDepartment of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, NorwayBackgroundStreptococcus dysgalactiae (SD) is a potent pathogen associated with infections in a broad range of host species. Notably, a substantial proportion of SD isolates exhibit reduced susceptibility to tetracycline but lack identifiable resistance determinants. In the present study, we wanted to explore the genetic basis for this low-grade resistance to tetracycline.MethodsGenome-wide association studies were performed on a collection of 407 SD genomes to identify potential novel resistance determinants. Two strains of SD, belonging to each of the subspecies dysgalactiae and equisimilis were used for mutagenesis. Natural transformation was exploited to knock out resistance gene candidates, and the resultant mutants were compared with their respective wildtypes regarding susceptibility to tetracycline, doxycycline, minocycline, tigecycline, erythromycin, gentamicin, clindamycin and ciprofloxacin.ResultsWe identified a two gene operon, herein designated trexAB, significantly associated with reduced susceptibility to tetracycline. The proteins encoded by the operon were predicted in silico to constitute a heterodimeric efflux transporter. The knockout of trexAB led to a 16- to 32-fold reduction in minimum inhibitory concentration (MIC) for tetracycline and a 4-fold reduction in MIC for tigecycline in the investigated strains. No differences between mutants and wildtypes were observed for other antibiotics included in the test panel. Whole genome alignment of mutants and their respective wildtypes revealed no differences other than the expected differences caused by the knockout.ConclusionWe have characterized a novel operon causing low-grade resistance to tetracycline in SD. The MIC distribution of trexAB-positive isolates is intersected by the current EUCAST susceptibility breakpoint, and our findings are relevant for future revisions and determinations of adequate breakpoints for tetracycline in S. dysgalactiae.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1583926/fullantibiotic resistancetetracyclineStreptococcus dysgalactiaenatural transformationABC transporter
spellingShingle Marte Glambek
Marte Glambek
Morten Kjos
Marita T. Mårli
Zhian Salehian
Steinar Skrede
Steinar Skrede
Audun Sivertsen
Bård R. Kittang
Bård R. Kittang
Oddvar Oppegaard
Oddvar Oppegaard
TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae
Frontiers in Cellular and Infection Microbiology
antibiotic resistance
tetracycline
Streptococcus dysgalactiae
natural transformation
ABC transporter
title TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae
title_full TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae
title_fullStr TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae
title_full_unstemmed TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae
title_short TrexAB, a novel tetracycline resistance determinant in Streptococcus dysgalactiae
title_sort trexab a novel tetracycline resistance determinant in streptococcus dysgalactiae
topic antibiotic resistance
tetracycline
Streptococcus dysgalactiae
natural transformation
ABC transporter
url https://www.frontiersin.org/articles/10.3389/fcimb.2025.1583926/full
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