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Home / Equine Research Bank / Animals (Basel) / Impact of Ambient Temperature Sample Storage on the Equine F...
Animals : an open access journal from MDPI2021; 11(3); 819; doi: 10.3390/ani11030819

Impact of Ambient Temperature Sample Storage on the Equine Fecal Microbiota.

Abstract: Sample storage conditions are an important factor in fecal microbiota analyses in general. The objective of this study was to investigate the effect of sample storage at room temperature on the equine fecal microbiota composition. Fecal samples were collected from 11 healthy horses. Each sample was divided into 7 sealed aliquots. One aliquot was immediately frozen at -80 °C; the remaining aliquots were stored at room temperature (21 to 22 °C) with one transferred to the freezer at each of the following time points: 6, 12, 24, 48, 72 and 96 h. The Illumina MiSeq sequencer was used for high-throughput sequencing of the V4 region of the 16S rRNA gene. Fibrobacteraceae () and Ruminococcaceae () were enriched in samples from 0 h and 6 h, whereas taxa from the families Bacillaceae, Planococcaceae, Enterobacteriaceae and Moraxellaceae were enriched in samples stored at room temperature for 24 h or greater. Samples frozen within the first 12 h after collection shared similar community membership. The community structure was similar for samples collected at 0 h and 6 h, but it was significantly different between samples frozen at 0 h and 12 h or greater. In conclusion, storage of equine fecal samples at ambient temperature for up to 6 h before freezing following sample collection had minimal effect on the microbial composition. Longer-term storage at ambient temperature resulted in alterations in alpha-diversity, community membership and structure and the enrichment of different taxa when compared to fecal samples immediately frozen at -80 °C.
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Publication Date: 2021-03-15 PubMed ID: 33803934PubMed Central: PMC8001224DOI: 10.3390/ani11030819Google Scholar: Lookup
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  • 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 article explores how the storage conditions of equine fecal samples, particularly when stored at room temperature, can affect the characteristics of the microbiota within the samples. The findings indicate storage at room temperature for up to 6 hours before freezing has minimal effect on the microbiota, however, longer-term storage leads to significant changes in the microbial composition.

Objective and Methodology

  • The objective of this study was to examine how the storage conditions of equine fecal samples at room temperature impact the fecal microbiota composition. This is significant as the accuracy of fecal microbiota analyses often hinges on the sample’s storage conditions.
  • Fecal samples were obtained from 11 healthy horses, each sample was divided into 7 sealed parts. One part was instantly frozen at -80 °C, and remaining parts were frozen after being kept at room temperature (21 to 22 °C) for 6, 12, 24, 48, 72 and 96 hours.
  • The V4 region of the 16S rRNA gene within each of the samples was sequenced using a high-throughput Illumina MiSeq sequencer for comprehensive analysis.

Findings

  • Families Fibrobacteraceae and Ruminococcaceae were enriched in samples frozen immediately and after 6 hours of storage at room temperature.
  • Taxa from the families Bacillaceae, Planococcaceae, Enterobacteriaceae, and Moraxellaceae showed an increased presence in samples stored at room temperature for 24 hours or more.
  • Fecal samples frozen within the first 12 hours of collection demonstrated similar microbial community membership. In contrast, the community structure was remarkably different between the samples frozen immediately and those frozen after 12 hours or more.

Conclusion

  • The study concludes that equine fecal samples can be stored at room temperature for up to 6 hours with minimal changes to the microbial composition. Meaning, freezing samples within this timeframe does not significantly alter the microbiota compared to those frozen immediately.
  • However, long-term storage at room temperature influences the alpha-diversity, community membership, and structure of the microbial composition in the samples. This underscores the importance of promptly freezing samples for in-depth and accurate fecal microbiota analyses.

Cite This Article

APA
Martin de Bustamante M, Plummer C, MacNicol J, Gomez D. (2021). Impact of Ambient Temperature Sample Storage on the Equine Fecal Microbiota. Animals (Basel), 11(3), 819. https://doi.org/10.3390/ani11030819

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 11
Issue: 3
PII: 819

Researcher Affiliations

Martin de Bustamante, Michelle
  • Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
Plummer, Caryn
  • Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
  • Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
MacNicol, Jennifer
  • Department of Animal Biosciences, Ontario Agriculture College, University of Guelph, Guelph, ON N1G 2W1, Canada.
Gomez, Diego
  • Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
  • Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, ON N1G 2W1, Canada.

Grant Funding

  • Intramural Grant / University of Florida College of Veterinary Medicine

Conflict of Interest Statement

The authors have no conflict of interest to declare.

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Citations

This article has been cited 18 times.
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