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Animals : an open access journal from MDPI2022; 12(15); 2009; doi: 10.3390/ani12152009

A Comparison of Methods to Maintain the Equine Cecal Microbial Environment In Vitro Utilizing Cecal and Fecal Material.

Abstract: The equine gastrointestinal (GI) microbiota is intimately related to the horse. The objective of the current study was to evaluate the microbiome and metabolome of cecal inoculum maintained in an anaerobic chamber or chemostat batch fermenter, as well as the fecal slurry maintained in an anaerobic chamber over 48 h. Cecal and fecal content were collected from healthy adult horses immediately upon death. Cecal fluid was used to inoculate chemostat vessels (chemostat cecal, = 11) and vessels containing cecal fluid (anaerobic cecal, = 15) or 5% fecal slurry (anaerobic fecal, = 6) were maintained in an anaerobic chamber. Sampling for microbiome and metabolome analysis was performed at vessel establishment (0 h), and after 24 h and 48 h of fermentation. Illumina sequencing was performed, and metabolites were identified via nuclear magnetic resonance (NMR). Alpha and beta diversity indices, as well as individual metabolite concentrations and metabolite regression equations, were analyzed and compared between groups and over time. No differences were evident between alpha or beta diversity in cecal fluid maintained in either an anaerobic chamber or chemostat. The microbiome of the fecal inoculum maintained anaerobically shifted over 48 h and was not comparable to that of the cecal inoculum. Metabolite concentrations were consistently highest in chemostat vessels and lowest in anaerobic fecal vessels. Interestingly, the rate of metabolite change in anaerobic cecal and chemostat cecal vessels was comparable. In conclusion, maintaining an equine cecal inoculum in either an anaerobic chamber or chemostat vessel for 48 h is comparable in terms of the microbiome. However, the microbiome and metabolome of fecal material is not comparable with a cecal inoculum. Future research is required to better understand the factors that influence the level of microbial activity in vitro, particularly when microbiome data identify analogous communities.
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Publication Date: 2022-08-08 PubMed ID: 35953998PubMed Central: PMC9367579DOI: 10.3390/ani12152009Google Scholar: Lookup
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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 study investigates different methods to maintain the microbial environment of the horse gastrointestinal system using both cecal and fecal materials in vitro, and compares the microbiome and metabolome in these two environments. The research concludes that while there are no significant differences between the various methods in terms of microbiome, there are significant differences in metabolite concentrations and the microbiome of fecal material does not match that of the cecal inoculum.

Research Context and Objectives

  • The study aimed to evaluate the microbiome (the combined genetic material of all the microorganisms in a particular environment) and metabolome (the total collection of the metabolites, which are small molecules, in a biological organism, system, or sample) of cecal inoculum maintained in an anaerobic chamber or chemostat batch fermenter, and the fecal slurry maintained in an anaerobic chamber over a 48-hour period.
  • The cecal and fecal contents used for the study were collected from healthy adult horses immediately after their death.

Methodology

  • The researchers used cecal fluid to inoculate chemostat vessels (chemostat’s cecal) and vessels containing cecal fluid (anaerobic cecal), while 5% of fecal slurry (anaerobic fecal) was maintained in an anaerobic chamber.
  • Sampling for microbial and metabolic analysis was done at vessel setup, and after 24 hours and 48 hours of fermentation. The research team carried out Illumina sequencing and identified metabolites via nuclear magnetic resonance (NMR).

Results and Analysis

  • The study found no differences in the alpha or beta diversity in cecal fluid maintained in either an anaerobic chamber or chemostat.
  • The microbiome of the fecal inoculum maintained anaerobically shifted over 48 hours and did not match that of the axial inoculum.
  • The metabolite concentrations consistently appeared highest in the chemostat vessels and lowest in the anaerobic fecal vessels. However, the rate of metabolite change in the anaerobic cecal and chemostat cecal vessels was the same.

Conclusions and Recommendations

  • The research concluded that maintaining equine cecal inoculum either in an anaerobic chamber or chemostat vessel for 48 hours is equivalent in terms of the presence and diversity of microbiome. Still, the microbiome and metabolome of fecal material do not coincide with the cecal inoculum.
  • The authors suggested that further research is required to better understand the factors influencing the level of microbial activity in vitro, especially when microbiome data show similar communities.’

Cite This Article

APA
MacNicol JL, Renwick S, Ganobis CM, Allen-Vercoe E, Weese JS, Pearson W. (2022). A Comparison of Methods to Maintain the Equine Cecal Microbial Environment In Vitro Utilizing Cecal and Fecal Material. Animals (Basel), 12(15), 2009. https://doi.org/10.3390/ani12152009

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 12
Issue: 15
PII: 2009

Researcher Affiliations

MacNicol, Jennifer L
  • Department of Animal Biosciences, Ontario Agricultural College, University of Guelph, Guelph, ON N1G 2W1, Canada.
Renwick, Simone
  • Department of Molecular and Cellular Biology, College of Biological Sciences, University of Guelph, Guelph, ON N1G 2W1, Canada.
Ganobis, Caroline M
  • Department of Molecular and Cellular Biology, College of Biological Sciences, University of Guelph, Guelph, ON N1G 2W1, Canada.
Allen-Vercoe, Emma
  • Department of Molecular and Cellular Biology, College of Biological Sciences, University of Guelph, Guelph, ON N1G 2W1, Canada.
Weese, Jeffery Scott
  • Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, ON N1G 2W1, Canada.
Pearson, Wendy
  • Department of Animal Biosciences, Ontario Agricultural College, University of Guelph, Guelph, ON N1G 2W1, Canada.

Grant Funding

  • N/A / Natural Sciences and Engineering Research Council
  • N/A / Selected Bioprodcts INC.

Conflict of Interest Statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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Citations

This article has been cited 2 times.
  1. Lagounova M, MacNicol JL, Weese JS, Pearson W. The Effect of Dietary Synbiotics in Actively Racing Standardbred Horses Receiving Trimethoprim/Sulfadiazine. Animals (Basel) 2023 Jul 18;13(14).
    doi: 10.3390/ani13142344pubmed: 37508120google scholar: lookup
  2. MacNicol JL, Renwick S, Ganobis CM, Allen-Vercoe E, Weese JS, Pearson W. The influence of a probiotic/prebiotic supplement on microbial and metabolic parameters of equine cecal fluid or fecal slurry in vitro. J Anim Sci 2023 Jan 3;101.
    doi: 10.1093/jas/skad034pubmed: 36715114google scholar: lookup
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