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PloS one2020; 15(1); e0227151; doi: 10.1371/journal.pone.0227151

Mapping the bacterial ecology on the phyllosphere of dry and post soaked grass hay for horses.

Abstract: Soaking hay fodder to reduce dust and soluble carbohydrate (WSC) contents prior to feeding is common practice among horse owners. Soaking can increase bacteria load in hay but no information exists on how this process alters the bacteria profile, which could pose a health risk or digestive challenge, to horses by introducing foreign bacteria into the gastrointestinal tract and so altering the normal profile. The current objectives were to map the bacterial profile of 3 different hays and determine how soaking alters this with the aim of improving best practice when feeding stabled horses. A Perennial Rye grass hay and two meadow s hays were soaked for 0, 1.5, 9 or 16 hours. Pre and post treatment, hays were analysed for WSC and total aerobic bacteria (CFU/g), and differences in bacteria family profiles were determined using ANOVA with significance set at P<0.05. Bacteria were identified via genomic DNA extraction and 16S library preparation (V3 and V4 variable region of 16S rRNA) according to the Illumina protocol. Differences in family operational taxonomic units (OTUs) between individual dry hays and different soaking times were identified via paired t-tests on the DESeq2 normalised data and false discovery rates accounted for using Padj (P<0.05). Mean % WSC losses and actual g/kg lost on DM basis (+/- SE) increased with soaking time being 18% = 30 (10.7), 38% = 72 (43.7), and 42% = 80 (38.8) for 1.5, 9 and 16 hours soak respectively. No relationship existed between WSC leaching and bacteria growth or profile. Grass type influenced bacterial profiles and their responses to soaking, but no differences were seen in richness or Shannon diversity indices. PCA analyses showed clustering of bacteria between meadow hays which differed from the perennial rye grass hay and this difference increased post soaking. Soaking hay pre-feeding causes inconsistent WSC leaching, bacteria growth and alterations in bacterial profiles which are unpredictable but may decrease the hygienic quality of the fodder.
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Publication Date: 2020-01-27 PubMed ID: 31986161PubMed Central: PMC6984722DOI: 10.1371/journal.pone.0227151Google 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.

This study examines how soaking hay, a widely-practiced method among horse owners to reduce dust and soluble carbohydrate content, affects the bacterial profile of the hay, potentially introducing foreign bacteria into horses’ digestive systems. The research aimed to map these bacterial changes in three types of hay soaked for varying duration and to understand how these changes might affect the feeding practices for stabled horses.

Methodology

  • Three types of hay, including Perennial Rye grass hay and two types of meadow hays, were soaked for 0, 1.5, 9, or 16 hours.
  • The study assessed the hay for soluble carbohydrate content and total aerobic bacteria before and after soaking.
  • Bacterial profiles were identified using genomic DNA extraction and a library preparation technique focused on regions of the 16S rRNA.
  • The study used an ANOVA test, and significance levels were set at P<0.05 to determine differences in bacteria family profiles.
  • The researchers also carried out paired t-tests on the DESeq2 normalised data to identify changes in bacterial units due to soaking time, accounting for false discovery rates.

Findings

  • The immersion of the hay resulted in increased soluble carbohydrate losses, with losses of 18%, 38% and 42% for soaking durations of 1.5, 9, and 16 hours respectively.
  • There was no observed relationship between soluble carbohydrate leaching and bacteria growth or profile.
  • Soaking the hay influenced bacterial profiles, with differing responses observed for different grass types.
  • No differences were observed in the richness or diversity (Shannon indices) of the bacteria due to soaking.
  • A specific clustering of bacteria was observed in meadow hays, differing from the Perennial Rye grass hay, and this difference increased after soaking.

Conclusion

  • The practice of soaking hay before feeding leads to inconsistent soluble carbohydrate leaching, bacteria growth, and changes in bacterial profiles that are hard to predict.
  • These changes may lower the hygienic quality of the fodder, suggesting a potential need for revised best practice guidelines for feeding stabled horses.

Cite This Article

APA
Moore-Colyer M, Longland A, Harris P, Zeef L, Crosthwaite S. (2020). Mapping the bacterial ecology on the phyllosphere of dry and post soaked grass hay for horses. PLoS One, 15(1), e0227151. https://doi.org/10.1371/journal.pone.0227151

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 15
Issue: 1
Pages: e0227151
PII: e0227151

Researcher Affiliations

Moore-Colyer, Meriel
  • School of Equine Management and Science, Royal Agricultural University, Cirencester, Gloucestershire, United Kingdom.
Longland, Annette
  • Equine and Livestock Nutrition Services, Tregaron, Ceredigion, Wales.
Harris, Patricia
  • Mars Horsecare United Kingdom LTD; Equine Studies Group, WALTHAM Centre for Pet Nutrition, Leicestershire, United Kingdom.
Zeef, Leo
  • Faculty of Biology, Medicine and Health, University of Manchester, United Kingdom.
Crosthwaite, Susan
  • NIAB, EMR, East Malling, Kent, United Kingdom.

MeSH Terms

  • Animal Feed / microbiology
  • Animals
  • Animals, Domestic
  • Bacteria, Aerobic / genetics
  • Bacteria, Aerobic / growth & development
  • Base Sequence
  • Carbohydrates / analysis
  • DNA, Bacterial / genetics
  • Horses
  • Lolium / microbiology
  • Polymerase Chain Reaction
  • Principal Component Analysis
  • RNA, Ribosomal, 16S
  • Water
  • Wettability

Conflict of Interest Statement

The commercial affiliation with Mars Horsecare for this study does not alter our adherence to PLOS ONE policies on sharing data and materials.

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Citations

This article has been cited 1 times.
  1. Daniels S, Hepworth J, Moore-Colyer M. The haybiome: Characterising the viable bacterial community profile of four different hays for horses following different pre-feeding regimens. PLoS One 2020;15(11):e0242373.
    doi: 10.1371/journal.pone.0242373pubmed: 33201929google scholar: lookup
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