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

Author:

Ward Ashley B.123ORCID,Harris Patricia A.4ORCID,Argo Caroline McG.1,Watson Christine5,Neacsu Madalina2,Russell Wendy R.2ORCID,Ribeiro Antonio36,Collie-Duguid Elaina36ORCID,Heidari Zeynab36,Morrison Philippa K.1ORCID

Affiliation:

1. School of Veterinary Medicine, Scotland’s Rural College, Aberdeen AB21 9YA, UK

2. The Rowett Institute, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK

3. School of Medicine Medical Sciences and Nutrition, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK

4. Equine Studies Group, Waltham Petcare Science Institute, Leicestershire LE14 4RT, UK

5. Department of Rural Land Use, Scotland’s Rural College, Aberdeen AB21 9YA, UK

6. Centre for Genome-Enabled Biology and Medicine, University of Aberdeen, King’s College, Aberdeen AB24 3FX, UK

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 n = 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 (p = 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.

Funder

Mars Petcare UK

Scottish Funding Council Research Excellence Grant

Rural and Environmental Sciences and Analytical Services Division

Publisher

MDPI AG

Subject

General Veterinary,Animal Science and Zoology

Reference51 articles.

1. Biddle, A., Nadeau, J., Reed, S., Ericsson, A., Coleman, R., Coleman, S., Wickens, C., Brooks, S., Hammer, C., and Mastellar, S. (2023, October 01). National Information Management and Support System. NE2202: The Equine Microbiome. Available online: https://www.nimss.org/projects/view/mrp/outline/18869.

2. Changes in Equine Hindgut Bacterial Populations during Oligofructose- Induced Laminitis;Milinovich;Environ. Microbiol.,2006

3. Dietary Fructan Carbohydrate Increases Amine Production in the Equine Large Intestine: Implications for Pasture-Associated Laminitis1;Crawford;J. Anim. Sci.,2007

4. Tuniyazi, M., He, J., Guo, J., Li, S., Zhang, N., Hu, X., and Fu, Y. (2021). Changes of Microbial and Metabolome of the Equine Hindgut during Oligofructose-Induced Laminitis. BMC Vet. Res., 17.

5. Fecal Microbiota of Horses with Colitis and Its Association with Laminitis and Survival during Hospitalization;Ayoub;Vet. Intern. Med.,2022

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3