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SVR-flaA typing of erythromycin- and ciprofloxacin-resistant Campylobacter jejuni strains isolated from poultry slaughterhouses in southern Brazil

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Abstract

The emergence of fluoroquinolone and macrolide resistance in C. jejuni, a recognized zoonotic pathogen, has increased worldwide. This study aimed to investigate phenotypic resistance to ciprofloxacin and erythromycin, the molecular mechanisms involved, and the strain of C. jejuni isolated from broiler carcasses. Eighty C. jejuni isolates from broiler carcasses in southern Brazil were investigated for their susceptibility to ciprofloxacin and erythromycin at minimal inhibitory concentrations. Mismatch amplification mutation assay–polymerase chain reaction (MAMA-PCR) was performed to detect substitutions of Thr-86-Ile, A2074C, and A2075G of domain V in the 23S rRNA. The presence of ermB gene and CmeABC operon were investigated by PCR. DNA sequencing was used to detect substitutions in the L4 and L22 proteins of the erythromycin-resistant strains. The Short Variable Region (SVR) of flaA was used to type all the strains resistant to both antimicrobials. Ciprofloxacin and erythromycin resistance were detected in 81.25% and 30.00% of the strains, respectively, and minimal inhibitory concentration values ranged from ≤ 0.125 to 64 µg/mL for ciprofloxacin and 0.5 to > 128 µg/mL for erythromycin. The Thr-86-Ile mutation in gyrA was observed in 100% of the ciprofloxacin-resistant strains. Mutations in both the A2074C and A2075G positions of 23S rRNA were observed in 62.5% of the erythromycin-resistant strains, while 37.5% had only the mutation A2075G. None of the strains harbored CmeABC operon, and ermB was not detected. Using DNA sequencing, the amino acid substitution T177S was detected in L4, and substitutions I65V, A103V, and S109A were detected in L22. Twelve flaA-SVR alleles were identified among the strains, with the most common SVR-flaA allele, type 287, covering 31.03% of ciprofloxacin- and erythromycin-resistant isolates. The present study revealed a high incidence and high levels of resistance to ciprofloxacin and erythromycin, as well as broad molecular diversity in C. jejuni isolates from broiler carcasses.

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Funding

This work was supported by the Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (E-26/010.001349/2019 and E-26/202.348/2022), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, code 001), and Conselho Nacional de Desenvolvimento Científico e Tecnológico.

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Authors and Affiliations

  1. Postgraduate Program in Veterinary Medicine (Veterinary Hygiene and Processing Technology of Animal Products), Faculdade de Veterinária, Universidade Federal Fluminense, Rua Vital Brasil Filho, 64, Zip Code: 24230340, Niteroi, RJ, Brazil

    Thomas Salles Dias, Arthur de Almeida Figueira, Gisllany Alves Costa, Virginia Léo de Almeida Pereira & Maria Helena Cosendey de Aquino

  2. Department of Preventive Veterinary Medicine, Faculdade de Veterinária, Universidade Federal Fluminense, Niteroi, RJ, Brazil

    Nathalie Costa da Cunha, Virginia Léo de Almeida Pereira & Maria Helena Cosendey de Aquino

  3. Laboratory of Molecular Epidemiology, Faculdade de Medicina Veterinária, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil

    Daise Aparecida Rossi & Roberta Torres de Melo

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  1. Thomas Salles Dias
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  2. Arthur de Almeida Figueira
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The original online version of this article was revised: In Conclusion section, the text "whole-genome sequencing was performed on strains circulating in the Brazilian poultry" needs to be corrected to "whole-genome sequencing should be performed on strains circulating in the Brazilian poultry"

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Dias, T.S., de Almeida Figueira, A., Costa, G.A. et al. SVR-flaA typing of erythromycin- and ciprofloxacin-resistant Campylobacter jejuni strains isolated from poultry slaughterhouses in southern Brazil. Braz J Microbiol 54, 1065–1073 (2023). https://doi.org/10.1007/s42770-023-00969-5

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