Shedding of Clostridium difficile PCR ribotype 078 by zoo animals, and report of an unstable metronidazole-resistant isolate from a zebra foal (Equus quagga burchellii).
Clostridium difficile is an emerging and potentially zoonotic pathogen, but its prevalence in most animal species, including exhibition animals, is currently unknown. In this study we assessed the prevalence of faecal shedding of C. difficile by zoo animals, and determined the ribotype, toxin profile and antimicrobial susceptibility of recovered isolates. A total of 200 samples from 40 animal species (36.5% of which came from plains zebra, Equus quagga burchellii) were analysed. C. difficile was isolated from 7 samples (3.5% of total), which came from the following animal species: chimpanzee (Pan troglodytes troglodytes), dwarf goat (Capra hircus), and Iberian ibex (Capra pyrenaica hispanica), with one positive sample each; and plains zebra, with 4 positive samples from 3 different individuals. Most recovered isolates (4/7, 57.1%) belonged to the epidemic PCR ribotype 078, produced toxins A and B, and had the genes encoding binary toxin (i.e. A(+)B(+)CDT(+) isolates). The remaining three isolates belonged to PCR ribotypes 039 (A(-)B(-)CDT(-)), 042 (A(+)B(+)CDT(-)) and 110 (A(-)B(+)CDT(-)). Regardless of their ribotype, all isolates displayed high-level resistance to the fluoroquinolones ciprofloxacin, enrofloxacin and levofloxacin. Some isolates were also resistant to meropenem and/or ertapenem. A ribotype 078 isolate recovered from a male zebra foal initially showed in vitro resistance to metronidazole (MIC ≥ 256 μg/ml), but lost that trait after subculturing on non-selective media. We conclude that zoo animals belonging to different species can carry ribotype 078 and other toxigenic strains of C. difficile showing resistance to antimicrobial compounds commonly used in veterinary and/or human medicine.
Department of Animal Health, Faculty of Veterinary, Universidad Complutense, Madrid, Spain; Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain.
Sergio
S
Álvarez-Pérez
Department of Animal Health, Faculty of Veterinary, Universidad Complutense, Madrid, Spain. Electronic address: jlblanco@vet.ucm.es.
José L
JL
Blanco
ZOO-AQUARIUM Madrid, Madrid, Spain.
Eva
E
Martínez-Nevado
Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain; Department of Medicine, Faculty of Medicine, Universidad Complutense, Madrid, Spain.
Teresa
T
Peláez
Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
Celine
C
Harmanus
Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
Ed
E
Kuijper
Department of Animal Health, Faculty of Veterinary, Universidad Complutense, Madrid, Spain.
Marta E
ME
García
D000818:Animals; D000836:Animals, Zoo; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D016360:Clostridioides difficile / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D024881:Drug Resistance, Bacterial; D004761:Enterocolitis, Pseudomembranous / Q000453:epidemiology / Q000662:veterinary*; D004768:Enterotoxins / Q000235:genetics; D018397:Equidae / Q000382:microbiology*; D005243:Feces / Q000382:microbiology; D024841:Fluoroquinolones / Q000494:pharmacology; D008297:Male; D008795:Metronidazole / Q000494:pharmacology*; D015995:Prevalence; D021521:Ribotyping
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; D024841:Fluoroquinolones; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile; D008795:Metronidazole
Clostridium difficile; Faecal shedding; Metronidazole resistance; Ribotype; Zebra; Zoo animals