Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.
Abstract: Although Clostridium difficile is recognized as a cause of enterocolitis in horses and humans, there has been little work published regarding the lability of C. difficile and its toxins in feces. A significant decrease in recovery of C. difficile from inoculated equine fecal samples occurred during storage. Recovery after storage in air at 4 degrees C decreased from 76% (37/49) after 24 hours to 67% (33/49) at 48 hours and 29% (14/ 49) after 72 hours. In contrast to aerobic storage, 25 of 26 samples stored anaerobically at 4 degrees C yielded growth of C. difficile for 30 days, whereas the organism was only detected for 2.5 +/- 2.52 days (x +/- SD) in paired samples stored aerobically. The use of an anaerobic transport medium was effective in maintaining viability of C. difficile. These findings indicate that poor aerotolerance is the reason for the rapid decrease in culture yield. In contrast to C. difficile organisms stored aerobically at 4 degrees C, C. difficile toxins were considerably more stable and could be detected by enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days. The poor survival of C. difficile but the stability of its toxins when feces are stored aerobically must be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses and when interpreting laboratory results.
Publication Date: 2000-07-25 PubMed ID: 10907862DOI: 10.1177/104063870001200406Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
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.
The research article explores the survival rate of the bacterium Clostridium difficile and its toxins in horse feces over time, under different conditions, and its implications for diagnostic methods and interpretation in cases of enterocolitis in horses and humans.
Decrease in C.difficile Recovery in Stored Samples
- The study found that the recovery rate, or the number of C. difficile bacteria that could be isolated from equine fecal samples, significantly decreased over time during storage. In the span of three days, the recovery rate from samples stored in air at 4 degrees Celsius decreased from 76% to 29%.
The Impact of Storage Conditions on C.difficile Growth
- In contrast to storage in air, almost all samples (25 out of 26) stored without exposure to air (anaerobically) at 4°C showed growth of C. difficile bacteria for a whole month. When stored with air (aerobically), the bacteria were only detected for an average of 2.5 days.
- The better survival rate of C. difficile under anaerobic storage conditions suggests that the bacterium has poor aerotolerance, or ability to survive in the presence of oxygen.
- An anaerobic transport medium was found effective in maintaining the viability of C. difficile, providing an important hint for better sample transportation.
Survival of C.difficile Toxins
- Despite the poor survival of C. difficile organisms in aerobic conditions, its toxins were significantly more stable. They could be identified using an enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days, regardless of the storage conditions.
Implications for Diagnostics and Interpretation
- The observed characteristics of C. difficile and its toxins need to be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses, as well as when interpreting the results. The findings stress the importance of quick sample processing or proper anaerobic storage/transportation to avoid false-negative culture results due to poor aerotolerance of the bacteria.
- Furthermore, the stable presence of C. difficile toxins implies that their detection can serve as a reliable diagnostic indicator for C. difficile-associated enterocolitis.
Cite This Article
APA
Weese JS, Staempfli HR, Prescott JF.
(2000).
Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.
J Vet Diagn Invest, 12(4), 332-336.
https://doi.org/10.1177/104063870001200406 Publication
Researcher Affiliations
- Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Canada.
MeSH Terms
- Animals
- Bacteria, Anaerobic
- Bacterial Toxins / pharmacology
- Clostridioides difficile
- Enterocolitis, Pseudomembranous / diagnosis
- Enterocolitis, Pseudomembranous / microbiology
- Enterocolitis, Pseudomembranous / veterinary
- Feces / microbiology
- Fetal Viability
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horses
- Survival Analysis
Citations
This article has been cited 18 times.- Redding LE, Daniel SG, Smith A, Keenan O, Barnhart D, Zackular JP. Comparison of Anaerobic Culture Methods for Detecting Clostridioides difficile in Bovine Faeces. Vet Med Sci 2025 Jan;11(1):e70085.
- Tyrnenopoulou P, Tsilipounidaki K, Florou Z, Gkountinoudis CG, Tyropoli K, Starras A, Peleki C, Marneris D, Arseniou N, Lianou DT, Katsarou EI, Petinaki E, Fthenakis GC. Detection of Gastrointestinal Pathogens with Zoonotic Potential in Horses Used in Free-Riding Activities during a Countrywide Study in Greece. Animals (Basel) 2024 Sep 3;14(17).
- Nho SW, Kim M, Kim SJ, Foley SL, Nayak R, Kweon O, Cerniglia CE. Pragmatic Strategy for Fecal Specimen Storage and the Corresponding Test Methods for Clostridioides difficile Diagnosis. Pathogens 2021 Aug 18;10(8).
- Kopper JJ, Willette JA, Kogan CJ, Seguin A, Bolin SR, Schott HC 2nd. Detection of pathogens in blood or feces of adult horses with enteric disease and association with outcome of colitis. J Vet Intern Med 2021 Sep;35(5):2465-2472.
- Tanaka M, Onizuka S, Mishima R, Nakayama J. Cultural isolation of spore-forming bacteria in human feces using bile acids. Sci Rep 2020 Sep 14;10(1):15041.
- Mallicote M, House AM, Sanchez LC. A review of foal diarrhoea from birth to weaning. Equine Vet Educ 2012 Apr;24(4):206-214.
- Grześkowiak Ł, Riedmüller J, Vahjen W, Zentek J. Storage procedures and time influence the detectability of Clostridium difficile toxin A but not toxin B in porcine fecal specimens. J Vet Diagn Invest 2020 Mar;32(2):222-225.
- Oliveira Júnior CA, Silva ROS, Cruz DSG, Pires IH, Alves GG, Lobato FCF. Evaluation of growth and sporulation of a non-toxigenic strain of Clostridioides difficile (Z31) and its shelf viability. Braz J Microbiol 2019 Jan;50(1):263-269.
- Andrés-Lasheras S, Martín-Burriel I, Mainar-Jaime RC, Morales M, Kuijper E, Blanco JL, Chirino-Trejo M, Bolea R. Preliminary studies on isolates of Clostridium difficile from dogs and exotic pets. BMC Vet Res 2018 Mar 9;14(1):77.
- Blanco N, Foxman B, Malani AN, Zhang M, Walk S, Rickard AH, Eisenberg MC. An in silico evaluation of treatment regimens for recurrent Clostridium difficile infection. PLoS One 2017;12(8):e0182815.
- Rodriguez-Palacios A, Stämpfli HR, Chang YF. False-Positive Clostridium difficile in Negative-Control Reactions Peak and Then Decrease with Repetitive Refrigeration of Immunoassay. Int Sch Res Notices 2014;2014:128120.
- Keessen EC, Hopman NE, van Leengoed LA, van Asten AJ, Hermanus C, Kuijper EJ, Lipman LJ. Evaluation of four different diagnostic tests to detect Clostridium difficile in piglets. J Clin Microbiol 2011 May;49(5):1816-21.
- Rodriguez-Palacios A, Lejeune JT. Moist-heat resistance, spore aging, and superdormancy in Clostridium difficile. Appl Environ Microbiol 2011 May;77(9):3085-91.
- Modi C, DePasquale JR, Nguyen NQ, Malinowski JE, Perez G. Does the handling time of unrefrigerated human fecal specimens impact the detection of Clostridium difficile toxins in a hospital setting?. Indian J Gastroenterol 2010 Jul;29(4):157-61.
- Bell JA, Kopper JJ, Turnbull JA, Barbu NI, Murphy AJ, Mansfield LS. Ecological characterization of the colonic microbiota of normal and diarrheic dogs. Interdiscip Perspect Infect Dis 2008;2008:149694.
- Jump RL, Pultz MJ, Donskey CJ. Vegetative Clostridium difficile survives in room air on moist surfaces and in gastric contents with reduced acidity: a potential mechanism to explain the association between proton pump inhibitors and C. difficile-associated diarrhea?. Antimicrob Agents Chemother 2007 Aug;51(8):2883-7.
- Weese JS, Armstrong J. Outbreak of Clostridium difficile-associated disease in a small animal veterinary teaching hospital. J Vet Intern Med 2003 Nov-Dec;17(6):813-6.
- Freeman J, Wilcox MH. The effects of storage conditions on viability of Clostridium difficile vegetative cells and spores and toxin activity in human faeces. J Clin Pathol 2003 Feb;56(2):126-8.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
