FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc2019; 32(2); 222-225; doi: 10.1177/1040638719864383

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.
Get Full Text
Publication Date: 2019-07-22 PubMed ID: 31328697PubMed Central: PMC7081514DOI: 10.1177/1040638719864383Google 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.
LinkedInCopy
  • Journal Article

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.

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

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
ISSN: 1943-4936
NlmUniqueID: 9011490
Country: United States
Language: English
Volume: 32
Issue: 2
Pages: 222-225

Researcher Affiliations

Grześkowiak, Łukasz
  • Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
Riedmüller, Jonathan
  • Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
Vahjen, Wilfried
  • Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
Zentek, Jürgen
  • 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.

References

This article includes 20 references
  1. Burnham CA, Carroll KC. Diagnosis of Clostridium difficile infection: an ongoing conundrum for clinicians and for clinical laboratories.. Clin Microbiol Rev 2013 Jul;26(3):604-30.
    pmc: PMC3719497pubmed: 23824374doi: 10.1128/cmr.00016-13google scholar: lookup
  2. Dice JF, Dehlinger PJ, Schimke RT. Studies on the correlation between size and relative degradation rate of soluble proteins.. J Biol Chem 1973 Jun 25;248(12):4220-8.
    pubmed: 4711605
  3. Drudy D, Fanning S, Kyne L. Toxin A-negative, toxin B-positive Clostridium difficile.. Int J Infect Dis 2007 Jan;11(1):5-10.
    pubmed: 16857405doi: 10.1016/j.ijid.2006.04.003google scholar: lookup
  4. 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.
    pmc: PMC1769877pubmed: 12560391doi: 10.1136/jcp.56.2.126google scholar: lookup
  5. Grześkowiak Ł, Zentek J, Vahjen W. Determination of the extent of Clostridium difficile colonisation and toxin accumulation in sows and neonatal piglets.. Anaerobe 2016 Aug;40:5-9.
    pubmed: 27108595doi: 10.1016/j.anaerobe.2016.04.012google scholar: lookup
  6. Grzeskowiak L, Martínez-Vallespín B, Dadi TH, Radloff J, Amasheh S, Heinsen FA, Franke A, Reinert K, Vahjen W, Zentek J, Pieper R. Formula Feeding Predisposes Neonatal Piglets to Clostridium difficile Gut Infection.. J Infect Dis 2018 Apr 11;217(9):1442-1452.
    pubmed: 29099941doi: 10.1093/infdis/jix567google scholar: lookup
  7. Grześkowiak Ł, Riedmüller J, de Thomasson H, Bordessoule S, Seyboldt C, Zentek J, Vahjen W. Porcine and bovine Clostridium difficile ribotype 078 isolates demonstrate similar growth and toxigenic properties.. Int Microbiol 2018 Dec;21(4):215-221.
    pubmed: 30810901doi: 10.1007/s10123-018-0018-xgoogle scholar: lookup
  8. Huang J, Yu Z, Gao H, Yan X, Chang J, Wang C, Hu J, Zhang L. Chemical structures and characteristics of animal manures and composts during composting and assessment of maturity indices.. PLoS One 2017;12(6):e0178110.
    pmc: PMC5467826pubmed: 28604783doi: 10.1371/journal.pone.0178110google scholar: lookup
  9. 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.
    pubmed: 20740339doi: 10.1007/s12664-010-0040-1google scholar: lookup
  10. Morris LS, Marchesi JR. Assessing the impact of long term frozen storage of faecal samples on protein concentration and protease activity.. J Microbiol Methods 2016 Apr;123:31-8.
    pmc: PMC4819717pubmed: 26853125doi: 10.1016/j.mimet.2016.02.001google scholar: lookup
  11. Moschonas G, Bolton DJ, McDowell DA, Sheridan JJ. Diversity of culturable psychrophilic and psychrotrophic anaerobic bacteria isolated from beef abattoirs and their environments.. Appl Environ Microbiol 2011 Jul;77(13):4280-4.
    pmc: PMC3127721pubmed: 21498765doi: 10.1128/aem.01778-10google scholar: lookup
  12. Podivinsky E, Love JL, van der Colff L, Samuel L. Effect of storage regime on the stability of DNA used as a calibration standard for real-time polymerase chain reaction.. Anal Biochem 2009 Nov 1;394(1):132-4.
    pubmed: 19549501doi: 10.1016/j.ab.2009.06.024google scholar: lookup
  13. Riegler M, Sedivy R, Pothoulakis C, Hamilton G, Zacherl J, Bischof G, Cosentini E, Feil W, Schiessel R, LaMont JT. Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro.. J Clin Invest 1995 May;95(5):2004-11.
    pmc: PMC295778pubmed: 7738167doi: 10.1172/jci117885google scholar: lookup
  14. Ross KS, Haites NE, Kelly KF. Repeated freezing and thawing of peripheral blood and DNA in suspension: effects on DNA yield and integrity.. J Med Genet 1990 Sep;27(9):569-70.
    pmc: PMC1017219pubmed: 2231649doi: 10.1136/jmg.27.9.569google scholar: lookup
  15. Rossmanith P, Röder B, Frühwirth K, Vogl C, Wagner M. Mechanisms of degradation of DNA standards for calibration function during storage.. Appl Microbiol Biotechnol 2011 Jan;89(2):407-17.
    pubmed: 20967442doi: 10.1007/s00253-010-2943-2google scholar: lookup
  16. Salnikova MS, Joshi SB, Rytting JH, Warny M, Middaugh CR. Physical characterization of clostridium difficile toxins and toxoids: effect of the formaldehyde crosslinking on thermal stability.. J Pharm Sci 2008 Sep;97(9):3735-52.
    pubmed: 18257030doi: 10.1002/jps.21261google scholar: lookup
  17. Schora DM, Peterson LR, Usacheva EA. Immunological Stability of Clostridium difficile Toxins in Clinical Specimens.. Infect Control Hosp Epidemiol 2018 Apr;39(4):434-438.
    pubmed: 29457584doi: 10.1017/ice.2018.20google scholar: lookup
  18. Silva ROS. Detection of enterotoxin A and cytotoxin B, and isolation of Clostridium difficile in piglets in Minas Gerais, Brazil. Ciência Rural 2011;41:1430–1435.
  19. Waters EH, Orr JP, Clark EG, Schaufele CM. Typhlocolitis caused by Clostridium difficile in suckling piglets.. J Vet Diagn Invest 1998 Jan;10(1):104-8.
    pubmed: 9526873doi: 10.1177/104063879801000122google scholar: lookup
  20. Weese JS, Staempfli HR, Prescott JF. Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.. J Vet Diagn Invest 2000 Jul;12(4):332-6.
    pubmed: 10907862doi: 10.1177/104063870001200406google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 100,129 horse owners like you who receive our email updates on horse health and nutrition.