Storage procedures and time influence the detectability of Clostridium difficile toxin A but not toxin B in porcine fecal specimens.
Abstract: Storage procedures are known to affect the detectability of toxins in equine and human feces. We assessed the impact of different storage conditions on the detectability of toxins in swine feces. Specimens were inoculated with toxins, 112 ng/g of toxin A (TcdA) and 16 ng/g of toxin B (TcdB) and subjected to the following 3 storage treatments: 4°C, -30°C, repetitive freezing at -30°C and thawing. Toxin determination was assessed at 1, 2, 7, 14, and 21 d with ELISA. A decrease in concentrations of TcdA with time was observed for samples stored at 4°C and repetitive freezing-thawing ( ≤0.05). On day 14, storage at 4°C resulted in decreased TcdA concentration as opposed to storage at -30°C and repetitive freezing-thawing ( ≤0.05). On day 21, storage at 4°C resulted in decreased TcdA detectability compared with storage at -30°C ( ≤0.05). The TcdB concentration was unaffected. These results on toxin detectability in swine feces should be carefully considered in in vitro studies on toxigenic Our results also offer valuable information for microbiologists and veterinarians monitoring the presence of virulent in pigs.
Publication Date: 2019-07-22 PubMed ID: 31328697PubMed Central: PMC7081514DOI: 10.1177/1040638719864383Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates how various storage procedures and times affect the detectability of two types of toxins, A and B, found in pig feces. The study found that while the detectability of toxin A was affected by storage, toxin B was not.
Methods
- The researchers performed their experiment using pig fecal specimens that had been inoculated with two different types of toxins, known as TcdA and TcdB.
- The toxin concentrations used were 112 nanograms per gram for TcdA, and 16 nanograms per gram for TcdB.
- The tests were carried out under three different storage conditions: 4°C, -30°C, and a repetitive cycle of freezing at -30°C and then thawing.
- The researchers then assessed the concentration of the toxins at various points in time: 1 day, 2 days, 7 days, 14 days, and 21 days after inoculation using an enzyme-linked immunosorbent assay (ELISA).
Findings
- The concentration of toxin A, TcdA, dropped over time in samples stored at 4°C as well as those subjected to a repeated cycle of freezing and thawing.
- By the 14th day, storage at 4°C resulted in a lower concentration of TcdA compared to storage at -30°C or the repetitive freezing-thawing.
- By the 21st day, it was harder to detect TcdA in samples stored at 4°C compared to those stored at -30°C.
- In contrast, the concentration of toxin B, TcdB, did not change regardless of the storage condition.
Implications
- The study’s findings on the detectability of these toxins in swine feces have important implications for future in vitro studies on toxicogenic C. difficile.
- They also provide vital information for microbiologists and veterinarians who need to monitor the presence of virulent C. difficile in pigs, as different storage conditions can affect the detectability of the bacteria’s toxins.
Cite This Article
APA
Grześkowiak Ł, Riedmüller J, Vahjen W, Zentek J.
(2019).
Storage procedures and time influence the detectability of Clostridium difficile toxin A but not toxin B in porcine fecal specimens.
J Vet Diagn Invest, 32(2), 222-225.
https://doi.org/10.1177/1040638719864383 Publication
Researcher Affiliations
- Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
- Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
- Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
- Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
MeSH Terms
- Animals
- Bacterial Proteins / isolation & purification
- Bacterial Toxins / isolation & purification
- Clostridioides difficile / isolation & purification
- Clostridium Infections / diagnosis
- Clostridium Infections / microbiology
- Clostridium Infections / veterinary
- Enterotoxins / isolation & purification
- Feces / microbiology
- Female
- Specimen Handling / methods
- Swine
- Swine Diseases / diagnosis
- Swine Diseases / microbiology
- Time Factors
Conflict of Interest Statement
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
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