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Molecular characterization of Clostridium difficile isolates from horses in an intensive care unit and association of disease severity with strain type

K. Gary MagdesianDepartment of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA 95616.

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Mauricio DujowichVeterinary Medical Teaching Hospital, School of Veterinary Medicine, University of California, Davis, CA 95616.

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John E. MadiganDepartment of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA 95616.

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Lori M. HansenCenter for Comparative Medicine, School of Medicine, University of California, Davis, CA 95616.

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Dwight C. HirshDepartment of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616.

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Spencer S. JangVeterinary Medical Teaching Hospital, School of Veterinary Medicine, University of California, Davis, CA 95616.

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Abstract

Objective—To determine molecular characteristics, antimicrobial susceptibility, and toxigenicity of Clostridium difficile isolates from horses in an intensive care unit and evaluate associations among severity of clinical disease with specific strains of C difficile.

Design—Prospective study.

Animals—130 horses.

Procedures—Feces were collected from horses admitted for acute gastrointestinal tract disease with loose feces and submitted for microbial culture and immunoassay for toxin production. Polymerase chain reaction assays were performed on isolates for toxins A and B genes and strain identification.

Results—Isolates were grouped into 3 strains (A, B, and C) on the basis of molecular banding patterns. Toxins A and B gene sequences were detected in 93%, 95%, and 73% of isolates of strains A, B, and C, respectively. Results of fecal immunoassays for toxin A were positive in 40%, 63%, and 16% of horses with strains A, B, and C, respectively. Isolates in strain B were resistant to metronidazole. Horses infected with strain B were 10 times as likely to have been treated with metronidazole prior to the onset of diarrhea as horses infected with other strains. Duration from onset of diarrhea to discharge (among survivors) was longer, systemic inflammatory response syndromes were more pronounced, and mortality rate was higher in horses infected with strain B than those infected with strains A and C combined.

Conclusions and Clinical Relevance—Horses may be infected with a number of heterogeneous isolates of C difficile. Results indicated that toxigenicity and antimicrobial susceptibility of isolates vary and that metronidazole-resistant strains may be associated with severe disease.

Abstract

Objective—To determine molecular characteristics, antimicrobial susceptibility, and toxigenicity of Clostridium difficile isolates from horses in an intensive care unit and evaluate associations among severity of clinical disease with specific strains of C difficile.

Design—Prospective study.

Animals—130 horses.

Procedures—Feces were collected from horses admitted for acute gastrointestinal tract disease with loose feces and submitted for microbial culture and immunoassay for toxin production. Polymerase chain reaction assays were performed on isolates for toxins A and B genes and strain identification.

Results—Isolates were grouped into 3 strains (A, B, and C) on the basis of molecular banding patterns. Toxins A and B gene sequences were detected in 93%, 95%, and 73% of isolates of strains A, B, and C, respectively. Results of fecal immunoassays for toxin A were positive in 40%, 63%, and 16% of horses with strains A, B, and C, respectively. Isolates in strain B were resistant to metronidazole. Horses infected with strain B were 10 times as likely to have been treated with metronidazole prior to the onset of diarrhea as horses infected with other strains. Duration from onset of diarrhea to discharge (among survivors) was longer, systemic inflammatory response syndromes were more pronounced, and mortality rate was higher in horses infected with strain B than those infected with strains A and C combined.

Conclusions and Clinical Relevance—Horses may be infected with a number of heterogeneous isolates of C difficile. Results indicated that toxigenicity and antimicrobial susceptibility of isolates vary and that metronidazole-resistant strains may be associated with severe disease.

Contributor Notes

Supported by the Center for Equine Health, with funding provided by the Oak Tree Racing Association, State of California pari-mutual wagering fund, and contributions from private donors.

Dr. Magdesian.