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Antibiotics (Basel, Switzerland)2025; 14(12); 1185; doi: 10.3390/antibiotics14121185

Antimicrobial-Resistant Enteric Gram-Negative Bacteria Isolated from a Fatal Diarrhea in a Horse: Genomic Characterization of CTX-M-2-Producing Escherichia coli.

Abstract: Infections caused by antimicrobial-resistant bacteria are difficult to treat and increase the risk of death in animals. This report describes a fatal case of diarrhea in a horse that, despite intensive treatment including surgery and broad-spectrum antimicrobials (ceftiofur and amikacin), experienced a worsening of its condition and subsequent death. A fecal swab sample was subjected to microbiological culture for the identification of bacteria and assessment of their phenotypical antimicrobial susceptibility profiles using the disk-diffusion and broth microdilution methods. The double-disk synergy test, polymerase chain reactions for the detection of genes encoding extended-spectrum β-lactamases, and whole-genome sequence-based analysis were also performed. Strains of and were isolated, with the strain DSL-HVUVV-2025 presenting resistance to a third-generation cephalosporin. Accordingly, the gene was identified in the DSL-HVUVV-2025 strain, which was submitted to whole-genome sequencing. Genomic analysis showed several antimicrobial resistance determinants, as well as virulence genes, including those associated with the enteroaggregative pathotype. The gene was surrounded by an IS element and embedded in a complex class 1 integron that is part of the Tn transposon. Strain DSL-HVUVV-2025 belonged to a novel sequence type. This case highlights the importance of monitoring antimicrobial resistance and performing genomic characterization of bacteria involved in equine diarrhea to guide effective clinical management in veterinary hospitals. It also reinforces the role of horses as potential carriers of WHO critical priority pathogens and the need for responsible antimicrobial use.
Publication Date: 2025-11-21 PubMed ID: 41463689PubMed Central: PMC12729381DOI: 10.3390/antibiotics14121185Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Antimicrobial-resistant bacteria caused a fatal diarrheal infection in a horse, and detailed genetic analysis identified a specific resistant Escherichia coli strain carrying CTX-M-2, highlighting the importance of monitoring and responsible antibiotic use in veterinary care.

Study Background and Objective

  • Infections by bacteria resistant to antimicrobial drugs pose treatment challenges and increase mortality in animals.
  • The study focuses on a fatal case of diarrhea in a horse that did not improve despite intensive treatment including surgery and broad-spectrum antibiotics (ceftiofur and amikacin).
  • The main goal was to identify the bacterial strains involved, characterize their antimicrobial resistance, and provide genomic insights into resistance mechanisms.

Methods Used in the Study

  • A fecal swab from the affected horse was collected for microbiological culture to isolate bacteria.
  • Phenotypical antimicrobial susceptibility testing was performed using:
    • Disk-diffusion method
    • Broth microdilution method
  • Double-disk synergy test was conducted to detect extended-spectrum β-lactamase (ESBL) production.
  • Polymerase chain reaction (PCR) was used to detect genes that encode ESBLs, focusing on the blaCTX-M-2 gene.
  • Whole-genome sequencing (WGS) was performed on the Escherichia coli strain to analyze resistance genes and virulence factors.

Key Findings

  • Multiple enteric Gram-negative bacteria strains were isolated; notable was the Escherichia coli strain DSL-HVUVV-2025.
  • This E. coli strain showed resistance to third-generation cephalosporins, important antibiotics in veterinary and human medicine.
  • The blaCTX-M-2 gene responsible for ESBL production was identified in this strain.
  • Genomic analysis revealed:
    • Multiple antimicrobial resistance genes implying multidrug resistance capabilities.
    • Virulence genes associated with the enteroaggregative E. coli pathotype.
    • The blaCTX-M-2 gene was located within a complex class 1 integron, which itself is part of a Tn transposon, indicating mobile genetic elements that facilitate spreading of resistance.
    • An insertion sequence (IS) element was surrounding the resistance gene, further highlighting the potential for gene mobility.
    • The strain belonged to a novel sequence type, suggesting it was a previously uncharacterized lineage.

Implications and Conclusions

  • This fatal case emphasizes the critical need for continuous surveillance of antimicrobial resistance in bacteria involved in equine infections.
  • Genomic characterization aids in understanding resistance mechanisms and guiding effective clinical management in veterinary settings.
  • Horses may serve as reservoirs or carriers of WHO critical priority pathogens, including multidrug-resistant bacteria.
  • There is a strong call for responsible antimicrobial use in veterinary medicine to prevent the emergence and spread of resistant pathogens.
  • The findings highlight the One Health concern, where antimicrobial resistance in animals can impact human and environmental health.

Cite This Article

APA
Rossi GAM, Sellera FP, Ferraz CM, Carvalho RS, Oliveira APL, Marques CA, Fávaro EBR, Rosa RDS, Silva LAM, Cardozo MV, Stehling EG, Furlan JPR. (2025). Antimicrobial-Resistant Enteric Gram-Negative Bacteria Isolated from a Fatal Diarrhea in a Horse: Genomic Characterization of CTX-M-2-Producing Escherichia coli. Antibiotics (Basel), 14(12), 1185. https://doi.org/10.3390/antibiotics14121185

Publication

ISSN: 2079-6382
NlmUniqueID: 101637404
Country: Switzerland
Language: English
Volume: 14
Issue: 12
PII: 1185

Researcher Affiliations

Rossi, Gabriel Augusto Marques
  • Department of Veterinary Medicine, University Vila Velha (UVV), Av. Comissário José Dantas de Melo, n.21, Vila Velha 29102-920, ES, Brazil.
Sellera, Fábio Parra
  • School of Veterinary Medicine, Metropolitan University of Santos, Santos 11045-002, SP, Brazil.
  • Department of Internal Medicine, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo 05508-220, SP, Brazil.
Ferraz, Carolina Magri
  • Department of Veterinary Medicine, University Vila Velha (UVV), Av. Comissário José Dantas de Melo, n.21, Vila Velha 29102-920, ES, Brazil.
Carvalho, Renan Silva de
  • Department of Veterinary Medicine, University Vila Velha (UVV), Av. Comissário José Dantas de Melo, n.21, Vila Velha 29102-920, ES, Brazil.
Oliveira, Alvaro de Paula Lage de
  • Department of Veterinary Medicine, University Vila Velha (UVV), Av. Comissário José Dantas de Melo, n.21, Vila Velha 29102-920, ES, Brazil.
Marques, Camila Angela
  • Department of Veterinary Medicine, University Vila Velha (UVV), Av. Comissário José Dantas de Melo, n.21, Vila Velha 29102-920, ES, Brazil.
Fávaro, Enzo Bernardes Rocha
  • Department of Veterinary Medicine, University Vila Velha (UVV), Av. Comissário José Dantas de Melo, n.21, Vila Velha 29102-920, ES, Brazil.
Rosa, Rafael da Silva
  • Department of Clinical Analyses, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto 19040-903, SP, Brazil.
Silva, Leandro Augusto Mariano
  • Department of Pathology, Reproduction and One Health, Faculty of Agricultural and Veterinary Sciences, São Paulo State University (UNESP), Jaboticabal 14884-900, SP, Brazil.
Cardozo, Marita Vedovelli
  • Department of Pathology, Reproduction and One Health, Faculty of Agricultural and Veterinary Sciences, São Paulo State University (UNESP), Jaboticabal 14884-900, SP, Brazil.
Stehling, Eliana Guedes
  • Department of Clinical Analyses, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto 19040-903, SP, Brazil.
Furlan, João Pedro Rueda
  • Department of Pharmaceutical Sciences, Health Sciences Center, Federal University of Paraíba, João Pessoa 58051-900, PB, Brazil.

Grant Funding

  • 139/2021 2021-WTG95 / FAPES
  • 23/12947-4 / FAPESP
  • 551/2023 P 2023-RH7P2 / FAPES
  • 304905/2022-4 / CNPq
  • 88887.824722/2023-00 / CAPES
  • Financial Code 001 / CAPES

Conflict of Interest Statement

The authors declare no conflicts of interest.

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