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Equine veterinary journal2013; 45(6); 671-675; doi: 10.1111/evj.12046

Detection of A/B toxin and isolation of Clostridium difficile and Clostridium perfringens from foals.

Abstract: Toxin detection and screening could contribute to knowledge of the transmission patterns, risk factors and epidemiology of Clostridium difficile and Clostridium perfringens. Objective: To isolate C. difficile and C. perfringens and to detect A/B toxins in faecal samples from diarrhoeic and nondiarrhoeic foals. Methods: Cross-sectional observational study. Methods: A total of 153 samples from foals were collected: 139 samples from farms and 14 samples from diarrhoeic foals admitted to a veterinary hospital. The A/B toxins were detected by cytotoxicity assay. All suspected colonies of C. perfringens were subjected to polymerase chain reaction for detection of the major toxin genes (α, β, ε and ι) and for detection of β2-, NetB- and enterotoxin-encoding genes. Furthermore, C. difficile and C. perfringens isolates were evaluated for in vitro antimicrobial susceptibility. Results: Seven of 153 (4.6%) samples, all from diarrhoeic foals, were positive for C. difficile A/B toxin. Of these, 5 of 14 (35.7%) were from hospitalised foals, and only 2 of 63 (3.2%) diarrhoeic foal samples were from farms (P = 0.002). Clostridium perfringens was isolated from 31 (20.3%) foals, of which 21 of 76 (27.6%) were diarrhoeic and 10 of 76 (13.2%) were nondiarrhoeic, demonstrating a difference between these 2 groups (P = 0.045). Only 4 strains were positive for the β2-encoding gene (cpb2). All C. difficile and C. perfringens isolates were susceptible to metronidazole and vancomycin. Conclusions: The present report highlights the need for laboratory diagnostics to differentiate C. difficile-associated infection in foals from other causes of diarrhoea to facilitate adequate antimicrobial therapy. Conclusions: More studies are needed to clarify the role of C. perfringens as a primary agent of diarrhoea in foals.
© 2013 EVJ Ltd.
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Publication Date: 2013-03-03 PubMed ID: 23452044DOI: 10.1111/evj.12046Google Scholar: Lookup
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  • Non-U.S. Gov't

Summary

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The research article details a study about the presence and role of Clostridium difficile and Clostridium perfringens, and their associated A/B toxins, in foals experiencing diarrhea. This study demonstrated a need for specific diagnostic procedures in distinguishing C. difficile infections in these animals to ensure appropriate treatments.

Research Objective and Method

  • The objective of this research was to isolate C. difficile and C. perfringens, and detect A/B toxins from the fecal samples of both diarrhoeic and nondiarrhoeic foals. The researchers aimed to enhance understanding of the transmission patterns, risk factors, and distribution of these bacteria.
  • The method followed a cross-sectional observational approach, with a total of 153 samples taken from foals. Of these, 139 samples originated from farms, and 14 were sourced from diarrhoeic foals admitted to a veterinary hospital for treatment.
  • Researchers used a cytotoxicity assay to detect the presence of A/B toxins. The study also employed polymerase chain reaction (PCR) to identify any significant toxin genes in suspected C. perfringens colonies.
  • Lastly, the researchers evaluated the antimicrobial susceptibility of C. difficile and C. perfringens isolates, specifically focusing on their sensitivity to metronidazole and vancomycin.

Research Findings

  • The experiment revealed that 4.6% of the total samples were positive for C. difficile A/B toxin, predominantly found in diarrhoeic foals. Strikingly, 35.7% of these were from hospitalised foals, while only 3.2% were sampled from farm animals.
  • The researchers isolated C. perfringens from 20.3% of the tested foals. This was split between diarrhoeic foals (27.6%) and nondiarrhoeic foals (13.2%), suggesting a correlation between the presence of this bacterium and symptoms of diarrhoea.
  • Testing identified the presence of the β2-encoding gene, cpb2, in only four C. perfringens strains.
  • Significantly, all isolated C. difficile and C. perfringens strains were revealed to be susceptible to metronidazole and vancomycin treatment.

Research Conclusions

  • The research concludes that there is a concrete need for laboratory diagnostics to distinguish C. difficile-associated infections in foals from other diarrhoea-causing factors, to ensure suitable antimicrobial therapy.
  • The researchers call for further studies to solidify the understanding of C. perfringens as a primary agent of diarrhoea in foals.

Cite This Article

APA
Silva RO, Ribeiro MG, Palhares MS, Borges AS, Maranhão RP, Silva MX, Lucas TM, Olivo G, Lobato FC. (2013). Detection of A/B toxin and isolation of Clostridium difficile and Clostridium perfringens from foals. Equine Vet J, 45(6), 671-675. https://doi.org/10.1111/evj.12046

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 45
Issue: 6
Pages: 671-675

Researcher Affiliations

Silva, R O S
  • Veterinary School, Universidade Federal de Minas Gerais (UFMG), Brazil.
Ribeiro, M G
    Palhares, M S
      Borges, A S
        Maranhão, R P A
          Silva, M X
            Lucas, T M
              Olivo, G
                Lobato, F C F

                  MeSH Terms

                  • Animals
                  • Bacterial Toxins
                  • Clostridioides difficile / isolation & purification
                  • Clostridium Infections / microbiology
                  • Clostridium Infections / veterinary
                  • Clostridium perfringens / genetics
                  • Clostridium perfringens / isolation & purification
                  • Clostridium perfringens / metabolism
                  • Cross-Sectional Studies
                  • Diarrhea / microbiology
                  • Diarrhea / veterinary
                  • Gene Expression Regulation, Bacterial / physiology
                  • Horse Diseases / microbiology
                  • Horses
                  • Polymerase Chain Reaction / veterinary

                  Citations

                  This article has been cited 19 times.
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                  5. Santana JA, Colombo SA, Silva BA, Diniz AN, de Almeida LR, Oliveira Junior CA, Lobato FCF, de Souza Trindade G, Paglia AP, Silva ROS. Clostridioides difficile and multi-drug-resistant staphylococci in free-living rodents and marsupials in parks of Belo Horizonte, Brazil. Braz J Microbiol 2022 Mar;53(1):401-410.
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                  6. Magdesian KG, Barnum S, Pusterla N. Fecal PCR testing for detection of Clostridium perfringens and Clostridioides difficile toxin genes and other pathogens in foals with diarrhea: 28 cases. J Vet Diagn Invest 2022 May;34(3):396-401.
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                  8. Ramos CP, Lopes EO, Oliveira Júnior CA, Diniz AN, Lobato FCF, Silva ROS. Immunochromatographic test and ELISA for the detection of glutamate dehydrogenase (GDH) and A/B toxins as an alternative for the diagnosis of Clostridioides (Clostridium) difficile-associated diarrhea in foals and neonatal piglets. Braz J Microbiol 2020 Sep;51(3):1459-1462.
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                  9. Viegas FM, Ramos CP, Xavier RGC, Lopes EO, Júnior CAO, Bagno RM, Diniz AN, Lobato FCF, Silva ROS. Fecal shedding of Salmonella spp., Clostridium perfringens, and Clostridioides difficile in dogs fed raw meat-based diets in Brazil and their owners' motivation. PLoS One 2020;15(4):e0231275.
                    doi: 10.1371/journal.pone.0231275pubmed: 32287295google scholar: lookup
                  10. Mehdizadeh Gohari I, Unterer S, Whitehead AE, Prescott JF. NetF-producing Clostridium perfringens and its associated diseases in dogs and foals. J Vet Diagn Invest 2020 Mar;32(2):230-238.
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                  12. Ramos CP, Santana JA, Morcatti Coura F, Xavier RGC, Leal CAG, Oliveira Junior CA, Heinemann MB, Lage AP, Lobato FCF, Silva ROS. Identification and Characterization of Escherichia coli, Salmonella Spp., Clostridium perfringens, and C. difficile Isolates from Reptiles in Brazil. Biomed Res Int 2019;2019:9530732.
                    doi: 10.1155/2019/9530732pubmed: 31263711google scholar: lookup
                  13. Silva ROS, de Oliveira Júnior CA, Blanc DS, Pereira ST, de Araujo MCR, Vasconcelos A, Lobato FCF. Clostridioides difficile infection in dogs with chronic-recurring diarrhea responsive to dietary changes. Anaerobe 2018 Jun;51:50-53.
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                  14. Diniz AN, Coura FM, Rupnik M, Adams V, Stent TL, Rood JI, de Oliveira CA Jr, Lobato FCF, Silva ROS. The incidence of Clostridioides difficile and Clostridium perfringens netF-positive strains in diarrheic dogs. Anaerobe 2018 Feb;49:58-62.
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                  15. Olivo G, Lucas TM, Borges AS, Silva RO, Lobato FC, Siqueira AK, da Silva Leite D, Brandão PE, Gregori F, de Oliveira-Filho JP, Takai S, Ribeiro MG. Enteric Pathogens and Coinfections in Foals with and without Diarrhea. Biomed Res Int 2016;2016:1512690.
                    doi: 10.1155/2016/1512690pubmed: 28116290google scholar: lookup
                  16. Finley A, Gohari IM, Parreira VR, Abrahams M, Staempfli HR, Prescott JF. Prevalence of netF-positive Clostridium perfringens in foals in southwestern Ontario. Can J Vet Res 2016 Jul;80(3):242-4.
                    pubmed: 27408339
                  17. Diniz AN, Silva ROS, Oliveira Junior CA, Pierezan F, Lobato FCF. Clostridium perfringens type A netF and netE positive and Clostridium difficile co-infection in two adult dogs. Anaerobe 2016 Apr;38:94-96.
                    doi: 10.1016/j.anaerobe.2015.12.013pubmed: 26762654google scholar: lookup
                  18. Silva RO, Rupnik M, Diniz AN, Vilela EG, Lobato FC. Clostridium difficile ribotypes in humans and animals in Brazil. Mem Inst Oswaldo Cruz 2015 Dec;110(8):1062-5.
                    doi: 10.1590/0074-02760150294pubmed: 26676318google scholar: lookup
                  19. Silva RO, Santos RL, Pires PS, Pereira LC, Pereira ST, Duarte MC, de Assis RA, Lobato FC. Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil. Braz J Microbiol 2013;44(1):133-7.
                    doi: 10.1590/S1517-83822013005000008pubmed: 24159295google scholar: lookup
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