Semi-quantitative analysis of Ruminococcus flavefaciens, Fibrobacter succinogenes and Streptococcus bovis in the equine large intestine using real-time polymerase chain reaction.
Abstract: There is a need to further our understanding of the role that the equine hindgut ecosystem plays in digestive processes and diseases. The aim of the present study was to utilise the real-time PCR technique to determine the abundance of candidate cellulolytic (Ruminococcus flavefaciens; Fibrobacter succinogenes) and non-cellulolytic (Streptococcus bovis) bacteria in lumen contents from the caecum, ventral and dorsal colon, and rectum of healthy horses (n 14). Total DNA was extracted from frozen and lyophilised lumen contents, and PCR primers and Taqman probes were designed based on 16S rDNA sequences for specific detection of candidate bacterial species. Overall, in frozen and lyophilised digesta, there were significantly (P F. succinogenes > S. bovis (P < 0.05), while in lyophilised digesta R. flavefaciens was present in significantly (P < 0.05) greater amounts than F. succinogenes and S. bovis. R. flavefaciens and F. succinogenes were abundant at significantly (P < 0.05) greater levels in lyophilised digesta v. frozen digesta, with no difference in S. bovis levels. These data indicate that for these bacteria at least, faeces are a suitable model for studying the bacterial ecosystem within the equine colon. The present study also indicates that the preservation method of digesta affects levels of bacteria detected.
Publication Date: 2008-04-01 PubMed ID: 18377691DOI: 10.1017/S0007114508968227Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper studies the presence and distribution of certain bacteria within the large intestine of horses, to further understand digestive processes and diseases among these creatures. The technique used is real-time PCR and the studied bacteria include Ruminococcus flavefaciens, Fibrobacter succinogenes and Streptococcus bovis.
Approach Used:
- The aim here is to determine the abundance of these bacteria in different parts of the horse’s large intestine – caecum, ventral and dorsal colon, as well as the rectum.
- This study was performed on healthy horses, 14 in number. Total DNA was extracted from the contents of these parts of the intestine, which was frozen and lyophilised (freeze-dried) prior to the procedure.
- The primers and Taqman probes for PCR were developed based on the 16S rDNA sequences of the bacteria under study. These tools allow for the specific detection of each type.
Results:
- In both frozen and lyophilised digesta there was a higher presence of Ruminococcus flavefaciens and Fibrobacter succinogenes compared to Streptococcus bovis. Notably, R. flavefaciens was found in even larger proportions than F. succinogenes.
- A significant finding is that there were greater levels of R. flavefaciens and F. succinogenes in lyophilised digesta compared to the frozen variety. The abundance of S. bovis did not show a difference between these two types of preparation.
Implications:
- These results suggest that studying faeces can be an effective approach to learning about the bacterial ecosystem within the equine colon, at least for these three kinds of bacteria.
- The study also indicates that the method of preserving the digesta plays a role in the levels of bacteria detected.
Cite This Article
APA
Hastie PM, Mitchell K, Murray JA.
(2008).
Semi-quantitative analysis of Ruminococcus flavefaciens, Fibrobacter succinogenes and Streptococcus bovis in the equine large intestine using real-time polymerase chain reaction.
Br J Nutr, 100(3), 561-568.
https://doi.org/10.1017/S0007114508968227 Publication
Researcher Affiliations
- Division of Veterinary Cell Sciences, Institute of Comparative Medicine, University of Glasgow Veterinary School, Bearsden Road, Glasgow G61 1QH, UK. P.Hastie@vet.gla.ac.uk
MeSH Terms
- Animals
- Cecum / microbiology
- Colon / microbiology
- Feces / microbiology
- Fibrobacter / genetics
- Horses / microbiology
- Intestine, Large / microbiology
- RNA, Ribosomal, 16S / analysis
- Rectum / microbiology
- Reverse Transcriptase Polymerase Chain Reaction / methods
- Ruminococcus / genetics
- Streptococcus bovis / genetics
Citations
This article has been cited 15 times.- Wunderlich G, Bull M, Ross T, Rose M, Chapman B. Understanding the microbial fibre degrading communities & processes in the equine gut. Anim Microbiome 2023 Jan 12;5(1):3.
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