In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance
The emergence of Neisseria gonorrhoeae strains resistant to extended-spectrum cephalosporins (ESCs) is a worldwide concern because this class of antibiotics represents the last empirical treatment option for gonorrhea. The abusive use of antimicrobials may be an essential factor for the emergence of...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-07-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.924764/full |
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| author | Marcos André Schörner Dany Mesa Fernando Hartmann Barazzetti Fernando Hartmann Barazzetti Jéssica Motta Martins Jéssica Motta Martins Hanalydia de Melo Machado Henrique Borges da Silva Grisard Julia Kinetz Wachter Márick Rodrigues Starick Márick Rodrigues Starick Mara Cristina Scheffer Jussara Kasuko Palmeiro Maria Luiza Bazzo Maria Luiza Bazzo Maria Luiza Bazzo |
| author_facet | Marcos André Schörner Dany Mesa Fernando Hartmann Barazzetti Fernando Hartmann Barazzetti Jéssica Motta Martins Jéssica Motta Martins Hanalydia de Melo Machado Henrique Borges da Silva Grisard Julia Kinetz Wachter Márick Rodrigues Starick Márick Rodrigues Starick Mara Cristina Scheffer Jussara Kasuko Palmeiro Maria Luiza Bazzo Maria Luiza Bazzo Maria Luiza Bazzo |
| author_sort | Marcos André Schörner |
| collection | DOAJ |
| description | The emergence of Neisseria gonorrhoeae strains resistant to extended-spectrum cephalosporins (ESCs) is a worldwide concern because this class of antibiotics represents the last empirical treatment option for gonorrhea. The abusive use of antimicrobials may be an essential factor for the emergence of ESC resistance in N. gonorrhoeae. Cephalosporin resistance mechanisms have not been fully clarified. In this study, we mapped mutations in the genome of N. gonorrhoeae isolates after resistance induction with cefixime and explored related metabolic pathways. Six clinical isolates with different antimicrobial susceptibility profiles and genotypes and two gonococcal reference strains (WHO F and WHO Y) were induced with increasing concentrations of cefixime. Antimicrobial susceptibility testing was performed against six antimicrobial agents before and after induction. Clinical isolates were whole-genome sequenced before and after induction, whereas reference strains were sequenced after induction only. Cefixime resistance induction was completed after 138 subcultures. Several metabolic pathways were affected by resistance induction. Five isolates showed SNPs in PBP2. The isolates M111 and M128 (ST1407 with mosaic penA-34.001) acquired one and four novel missense mutations in PBP2, respectively. These isolates exhibited the highest minimum inhibitory concentration (MIC) for cefixime among all clinical isolates. Mutations in genes contributing to ESC resistance and in other genes were also observed. Interestingly, M107 and M110 (ST338) showed no mutations in key determinants of ESC resistance despite having a 127-fold increase in the MIC of cefixime. These findings point to the existence of different mechanisms of acquisition of ESC resistance induced by cefixime exposure. Furthermore, the results reinforce the importance of the gonococcal antimicrobial resistance surveillance program in Brazil, given the changes in treatment protocols made in 2017 and the nationwide prevalence of sequence types that can develop resistance to ESC. |
| first_indexed | 2024-04-12T08:05:53Z |
| format | Article |
| id | doaj.art-9785f5c7ae494a5bbef9fc47a547531e |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-12T08:05:53Z |
| publishDate | 2022-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-9785f5c7ae494a5bbef9fc47a547531e2022-12-22T03:41:11ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-07-011210.3389/fcimb.2022.924764924764In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistanceMarcos André Schörner0Dany Mesa1Fernando Hartmann Barazzetti2Fernando Hartmann Barazzetti3Jéssica Motta Martins4Jéssica Motta Martins5Hanalydia de Melo Machado6Henrique Borges da Silva Grisard7Julia Kinetz Wachter8Márick Rodrigues Starick9Márick Rodrigues Starick10Mara Cristina Scheffer11Jussara Kasuko Palmeiro12Maria Luiza Bazzo13Maria Luiza Bazzo14Maria Luiza Bazzo15Laboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilInstituto de Pesquisa Pelé Pequeno Príncipe, Curitiba, PR, BrazilLaboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Bioinformática, Programa de Pós-Graduação em Biotecnologia e Biociências, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilPrograma de Pós-Graduação em Farmácia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilPrograma de Pós-Graduação em Farmacologia, Departamento de Farmacologia, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Imunofarmacologia e Doenças Infecciosas, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Microbiologia, Unidade do Laboratório de Análises Clínicas, Hospital Universitário Professor Polydoro Ernani de São Thiago, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilCentro de Ciências da Saúde, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilLaboratório de Biologia Molecular, Microbiologia e Sorologia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilPrograma de Pós-Graduação em Farmácia, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilCentro de Ciências da Saúde, Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, BrazilThe emergence of Neisseria gonorrhoeae strains resistant to extended-spectrum cephalosporins (ESCs) is a worldwide concern because this class of antibiotics represents the last empirical treatment option for gonorrhea. The abusive use of antimicrobials may be an essential factor for the emergence of ESC resistance in N. gonorrhoeae. Cephalosporin resistance mechanisms have not been fully clarified. In this study, we mapped mutations in the genome of N. gonorrhoeae isolates after resistance induction with cefixime and explored related metabolic pathways. Six clinical isolates with different antimicrobial susceptibility profiles and genotypes and two gonococcal reference strains (WHO F and WHO Y) were induced with increasing concentrations of cefixime. Antimicrobial susceptibility testing was performed against six antimicrobial agents before and after induction. Clinical isolates were whole-genome sequenced before and after induction, whereas reference strains were sequenced after induction only. Cefixime resistance induction was completed after 138 subcultures. Several metabolic pathways were affected by resistance induction. Five isolates showed SNPs in PBP2. The isolates M111 and M128 (ST1407 with mosaic penA-34.001) acquired one and four novel missense mutations in PBP2, respectively. These isolates exhibited the highest minimum inhibitory concentration (MIC) for cefixime among all clinical isolates. Mutations in genes contributing to ESC resistance and in other genes were also observed. Interestingly, M107 and M110 (ST338) showed no mutations in key determinants of ESC resistance despite having a 127-fold increase in the MIC of cefixime. These findings point to the existence of different mechanisms of acquisition of ESC resistance induced by cefixime exposure. Furthermore, the results reinforce the importance of the gonococcal antimicrobial resistance surveillance program in Brazil, given the changes in treatment protocols made in 2017 and the nationwide prevalence of sequence types that can develop resistance to ESC.https://www.frontiersin.org/articles/10.3389/fcimb.2022.924764/fullSNPwhole-genome sequencingPBP2penAresistance inductionmutations |
| spellingShingle | Marcos André Schörner Dany Mesa Fernando Hartmann Barazzetti Fernando Hartmann Barazzetti Jéssica Motta Martins Jéssica Motta Martins Hanalydia de Melo Machado Henrique Borges da Silva Grisard Julia Kinetz Wachter Márick Rodrigues Starick Márick Rodrigues Starick Mara Cristina Scheffer Jussara Kasuko Palmeiro Maria Luiza Bazzo Maria Luiza Bazzo Maria Luiza Bazzo In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance Frontiers in Cellular and Infection Microbiology SNP whole-genome sequencing PBP2 penA resistance induction mutations |
| title | In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance |
| title_full | In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance |
| title_fullStr | In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance |
| title_full_unstemmed | In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance |
| title_short | In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance |
| title_sort | in vitro selection of neisseria gonorrhoeae unveils novel mutations associated with extended spectrum cephalosporin resistance |
| topic | SNP whole-genome sequencing PBP2 penA resistance induction mutations |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.924764/full |
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