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Animals : an open access journal from MDPI2023; 13(19); 3107; doi: 10.3390/ani13193107

Homemade Nucleic Acid Preservation Buffer Proves Effective in Preserving the Equine Faecal Microbiota over Time at Ambient Temperatures.

Abstract: The equine faecal microbiota is often assessed as a proxy of the microbial community in the distal colon, where the microbiome has been linked to states of health and disease in the horse. However, the microbial community structure may change over time if samples are not adequately preserved. This study stored equine faecal samples from = 10 horses in four preservation treatments at room temperature for up to 150 h and assessed the resulting impact on microbial diversity and the differential abundance of taxa. Treatments included "COLD" (samples packaged with a cool pack), "CLX" (2% chlorhexidine digluconate solution), "NAP" (nucleic acid preservation buffer), and "FTA" (Whatman FTA™ cards). The samples were assessed using 16S rRNA gene sequencing after storage for 0, 24, 72, and 150 h at room temperature under the different treatments. The results showed effective preservation of diversity and community structure with NAP buffer but lower diversity ( = 0.001) and the under-representation of Fibrobacterota in the FTA card samples. The NAP treatment inhibited the overgrowth of bloom taxa that occurred by 72 h at room temperature. The COLD, CLX, and NAP treatments were effective in preserving the faecal microbiota for up to 24 h at room temperature, and the CLX and NAP treatments improved the yield of Patescibacteria and Fibrobacterota in some cases. The cold and CLX treatments were ineffective in preventing community shifts that occurred by 72 h at room temperature. These findings demonstrate the suitability of the COLD, NAP, and CLX treatments for the room temperature storage of equine faeces for up to 24 h and of NAP buffer for up to 150 h prior to processing.
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Publication Date: 2023-10-05 PubMed ID: 37835713PubMed Central: PMC10572018DOI: 10.3390/ani13193107Google 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.

The study investigated four preservation methods for horse fecal samples at room temperature and found that homemade nucleic acid preservation buffer was able to effectively preserve microbial diversity and structure over an extended time period.

Research Goals

In medical and veterinary sciences, fecal samples are often used to analyze the microbial community within an organism, which can help assess its health or diagnose diseases. However, these samples can degrade if not properly preserved, affecting the accuracy of results. This study aimed to analyze and compare the effectiveness of four preservation treatments for horse fecal samples:

  • “COLD” – storage with a cool pack
  • “CLX” – treatment with 2% chlorhexidine digluconate solution
  • “NAP” – treatment with nucleic acid preservation buffer
  • “FTA” – storage on Whatman FTA™ cards

Methodology

Horse fecal samples were taken from ten horses and subjected to the four preservation treatments. These were stored at room temperature for varying periods up to 150 hours. Using 16S rRNA gene sequencing, the research team analyzed the effect of the treatments over time on both microbial diversity and the differential abundance of taxa.

Results and Conclusion

The study found that, of the four treatments, NAP (nucleic acid preservation buffer) was the most effective at preserving diversity and community structure over a lengthy period. Specifically, it prevented the overgrowth of bloom taxa that happened by 72 hours at room temperature. While the treatments using a cool pack (COLD) and chlorhexidine digluconate (CLX) were efficient in preserving the fecal samples up to 24 hours, they didn’t prevent community changes that appeared by 72 hours in the room temperature. The Whatman FTA™ card method (FTA) was associated with reduced diversity and a lowered presence of Fibrobacterota.
Thus, the research concluded that for the short-term preservation of equine fecal samples (up to 24 hours), the COLD, NAP, and CLX methods are suitable. However, for longer-term preservation (up to 150 hours), NAP buffer is the recommended method.

Cite This Article

APA
Ward AB, Harris PA, Argo CM, Watson C, Neacsu M, Russell WR, Ribeiro A, Collie-Duguid E, Heidari Z, Morrison PK. (2023). Homemade Nucleic Acid Preservation Buffer Proves Effective in Preserving the Equine Faecal Microbiota over Time at Ambient Temperatures. Animals (Basel), 13(19), 3107. https://doi.org/10.3390/ani13193107

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 13
Issue: 19
PII: 3107

Researcher Affiliations

Ward, Ashley B
  • School of Veterinary Medicine, Scotland's Rural College, Aberdeen AB21 9YA, UK.
  • The Rowett Institute, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
  • School of Medicine Medical Sciences and Nutrition, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
Harris, Patricia A
  • Equine Studies Group, Waltham Petcare Science Institute, Leicestershire LE14 4RT, UK.
Argo, Caroline McG
  • School of Veterinary Medicine, Scotland's Rural College, Aberdeen AB21 9YA, UK.
Watson, Christine
  • Department of Rural Land Use, Scotland's Rural College, Aberdeen AB21 9YA, UK.
Neacsu, Madalina
  • The Rowett Institute, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
Russell, Wendy R
  • The Rowett Institute, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
Ribeiro, Antonio
  • School of Medicine Medical Sciences and Nutrition, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
  • Centre for Genome-Enabled Biology and Medicine, University of Aberdeen, King's College, Aberdeen AB24 3FX, UK.
Collie-Duguid, Elaina
  • School of Medicine Medical Sciences and Nutrition, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
  • Centre for Genome-Enabled Biology and Medicine, University of Aberdeen, King's College, Aberdeen AB24 3FX, UK.
Heidari, Zeynab
  • School of Medicine Medical Sciences and Nutrition, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
  • Centre for Genome-Enabled Biology and Medicine, University of Aberdeen, King's College, Aberdeen AB24 3FX, UK.
Morrison, Philippa K
  • School of Veterinary Medicine, Scotland's Rural College, Aberdeen AB21 9YA, UK.

Grant Funding

  • Mars Petcare UK / Mars Petcare UK
  • Scottish Funding Council Research Excellence Grant (REG) / Scottish Funding Council Research Excellence Grant (REG)

Conflict of Interest Statement

The authors declare no conflicts of interest. With the exception of PH (employed by the funding organization), the funding organization did not have any additional role in the conceptualization, methodology, investigation, data curation, formal analysis, decision to publish, or preparation of the manuscript. PH was involved in study design, data interpretation, and manuscript preparation.

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Citations

This article has been cited 3 times.
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