Abstract
BackgroundEvaluation of factors that might impact microbiota assessment is important to avoid spurious results, particularly in field and multicenter studies where sample collection may occur distant from the laboratory. This study evaluated the impact of refrigeration on next generation sequence-based assessment of the canine and feline fecal microbiota. Fecal samples were collected from seven dogs and ten cats, and analysed at baseline and after 3, 7, 10 and 14 days of storage at 4°C.ResultsThere were no differences in community membership or population structure between timepoints for either dogs or cats, nor were there any differences in richness, diversity and evenness. There were few differences in relative abundance of phyla or predominant genera, with the only differences being significant increases in Actinobacteria between Days 0-14 (P = 0.0184) and 1-14 (P = 0.0182) for canine samples, and a decrease in Erysipelotrichaceae incertae sedis between Day 0 and Day 7 (median 4.9 vs 2.2%, P = 0.046) in feline samples.Linear discriminant analysis effect size and indicator analysis identified a small number of genera that were over-represented in, or defining characteristics of, Day 14 samples. These were predominantly Proteobacteria and Actinobacteria, with Psychrobacter and Arthrobacter enriched in both canine and feline Day 14 samples.ConclusionsStorage for at least 14 days at 4°C has limited impact on culture-independent assessment of the canine and feline fecal microbiota, although changes in some individual groups may occur.
Highlights
Evaluation of factors that might impact microbiota assessment is important to avoid spurious results, in field and multicenter studies where sample collection may occur distant from the laboratory
A study of human and soil samples detected no influence of storage, including refrigeration, on microbial population structure or diversity, but some shifts in relative abundances of different taxa [8], results that were similar to a later study of soil bacteria [9]
Dogs were clinically normal with no history of antimicrobial exposure or gastrointestinal disease, medical histories were limited based on the nature of the population
Summary
Evaluation of factors that might impact microbiota assessment is important to avoid spurious results, in field and multicenter studies where sample collection may occur distant from the laboratory. This study evaluated the impact of refrigeration on generation sequence-based assessment of the canine and feline fecal microbiota. There has been limited study of the impact of storage on the fecal microbiota. A study of human fecal samples reported limited impact of storage at room temperature for 24 h or at -80°C for six months [6]. A study of human and soil samples detected no influence of storage, including refrigeration, on microbial population structure or diversity, but some shifts in relative abundances of different taxa [8], results that were similar to a later study of soil bacteria [9]
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