Effectiveness of stabilization methods for the immediate and short-term preservation of bovine fecal and upper respiratory tract genomic DNA.

Autor: Pinnell LJ; Veterinary Education, Research and Outreach Program, Texas A&M University, Canyon, TX, United States of America., Wolfe CA; Veterinary Education, Research and Outreach Program, Texas A&M University, Canyon, TX, United States of America., Castle J; Veterinary Education, Research and Outreach Program, Texas A&M University, Canyon, TX, United States of America., Crosby WB; Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Starkville, Mississippi State, United States of America., Doster E; Veterinary Education, Research and Outreach Program, Texas A&M University, Canyon, TX, United States of America., Morley PS; Veterinary Education, Research and Outreach Program, Texas A&M University, Canyon, TX, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2024 Apr 02; Vol. 19 (4), pp. e0300285. Date of Electronic Publication: 2024 Apr 02 (Print Publication: 2024).
DOI: 10.1371/journal.pone.0300285
Abstrakt: Previous research on stabilization methods for microbiome investigations has largely focused on human fecal samples. There are a few studies using feces from other species, but no published studies investigating preservation of samples collected from cattle. Given that microbial taxa are differentially impacted during storage it is warranted to study impacts of preservation methods on microbial communities found in samples outside of human fecal samples. Here we tested methods of preserving bovine fecal respiratory specimens for up to 2 weeks at four temperatures (room temperature, 4°C, -20°C, and -80°C) by comparing microbial diversity and community composition to samples extracted immediately after collection. Importantly, fecal specimens preserved and analyzed were technical replicates, providing a look at the effects of preservation method in the absence of biological variation. We found that preservation with the OMNIgene®•GUT kit resulted in community structure most like that of fresh samples extracted immediately, even when stored at room temperature (~20°C). Samples that were flash-frozen without added preservation solution were the next most representative of original communities, while samples preserved with ethanol were the least representative. These results contradict previous reports that ethanol is effective in preserving fecal communities and suggest for studies investigating cattle either flash-freezing of samples without preservative or preservation with OMNIgene®•GUT will yield more representative microbial communities.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Pinnell et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje