FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / PLoS One / Use of next generation sequencing to investigate the microbi...
PloS one2018; 13(11); e0206989; doi: 10.1371/journal.pone.0206989

Use of next generation sequencing to investigate the microbiota of experimentally induced wounds and the effect of bandaging in horses.

Abstract: To use next generation sequencing to characterize the microbiota of horses during healing of skin wounds in two anatomical locations (body and limb) known to present different healing patterns; and to investigate the impact of bandaging on bacterial communities of skin wounds located on the limbs of horses. Full-thickness skin wounds were created on the distal extremity of both thoracic limbs and on one lateral mid-thoracic wall of four healthy horses. Limb wounds were randomly assigned to bandaging or not. A full-thickness sample was collected with a biopsy punch from intact thorax and limb skin (T0) and from the margin of one wound per site (thorax, unbandaged limb, bandaged limb) 1 week (T1) and 2 weeks (T2) postoperatively, and at full healing (T3). Thoracic skin samples obtained from three healthy horses were included in the analysis as controls. Anatomic location (thorax vs. limb) significantly influenced bacterial composition of equine skin and healing wounds. Fusobacterium and Actinobacillus were strongly associated with limb wounds during the initial phases of healing. Bandaging had a significant impact on the microbiota during the healing process. The skin microbiota after healing was more similar to samples from controls, demonstrating the resilience and stability of the environment. Equine skin microbiota is a rich and stable environment that is disturbed by wounding, but returns to its previous stage after full healing. Anatomic location significantly influences bacterial composition of the equine skin during wound healing. Bandaging has a significant impact on the skin microbiota of horses during the healing process. Results of this study provide new insight for a better understanding of the contribution of bacteria to wound healing in horses and may facilitate the future development of therapeutic strategies using commensal bacteria.
Get Full Text
Publication Date: 2018-11-26 PubMed ID: 30475922PubMed Central: PMC6261015DOI: 10.1371/journal.pone.0206989Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 research article investigates the impact of wounding and bandaging on the skin microbiota of horses, and how these factors vary depending on the anatomical location of the wound.

Overview of the Research

  • This research used next generation sequencing to characterise the microbiota (bacterial communities) of horses during the healing process of skin wounds at two different locations, the body and limb. These two places are known to have different healing patterns.
  • The study included creating full-thickness skin wounds on the limbs and mid-thoracic wall of four healthy horses. Some limb wounds were bandaged, while some were left as is.
  • Full-thickness biopsy samples were then collected from intact skin and from the margin of one wound per site at different timeframes- postoperatively after 1 week, 2 weeks and at full healing.
  • The thoracic skin samples of three healthy horses were also included as control in the study.

Anatomical Location and Bacterial Composition

  • The research found that the anatomical location of the wound (thorax vs. limb) significantly influenced the bacterial composition of the equine skin and healing wounds.
  • Fusobacterium and Actinobacillus were found to be strongly associated with limb wounds during the initial phases of healing.

Impact of Bandaging

  • Bandaging of wounds also had a significant impact on the microbiota during the healing process, affecting the bacterial composition.
  • The skin microbiota after healing was found to be more similar to the samples from the control horses. This observation underlines the resilience and stability of the bacterial environment, confirming that it is disturbed by wounding but returns to its previous state after full healing.

Conclusions

  • The research concluded that the equine skin microbiota is a rich and stable environment that is affected by wounding, but reverts to its original state post healing. The location of the wound and bandaging also significantly impacts the bacterial composition of the equine skin during wound healing.
  • The findings could be instrumental in developing better therapeutic strategies using commensal bacteria, which could potentially enhance the wound healing process in horses.

Cite This Article

APA
Kamus LJ, Theoret C, Costa MC. (2018). Use of next generation sequencing to investigate the microbiota of experimentally induced wounds and the effect of bandaging in horses. PLoS One, 13(11), e0206989. https://doi.org/10.1371/journal.pone.0206989

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 13
Issue: 11
Pages: e0206989

Researcher Affiliations

Kamus, Louis J
  • Department of Veterinary Biomedical Sciences, University of Montreal, Saint-Hyacinthe, Q, Canada.
Theoret, Christine
  • Department of Veterinary Biomedical Sciences, University of Montreal, Saint-Hyacinthe, Q, Canada.
Costa, Marcio C
  • Department of Veterinary Biomedical Sciences, University of Montreal, Saint-Hyacinthe, Q, Canada.

MeSH Terms

  • Actinobacillus / genetics
  • Actinobacillus / isolation & purification
  • Animals
  • Bacteria / genetics
  • Bacteria / isolation & purification
  • Bandages
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism
  • Fusobacterium / genetics
  • Fusobacterium / isolation & purification
  • High-Throughput Nucleotide Sequencing
  • Horses
  • Microbiota
  • Principal Component Analysis
  • Sequence Analysis, DNA
  • Skin / microbiology
  • Skin / pathology
  • Skin Diseases / microbiology
  • Skin Diseases / pathology
  • Skin Diseases / veterinary
  • Wounds and Injuries / microbiology
  • Wounds and Injuries / pathology
  • Wounds and Injuries / veterinary

Conflict of Interest Statement

The authors have declared that no competing interests exist.

References

This article includes 42 references
  1. NAHMS: (November 2006) Part I: baseline reference of equine health and management. https://www.aphis.usda.gov/aniaml_health/nahms/equine/downloads/equine05/Equine05_dr_PartI.pdf
  2. Owen KR, Singer ER, Clegg PD, Ireland JL, Pinchbeck GL. Identification of risk factors for traumatic injury in the general horse population of north-west England, Midlands and north Wales.. Equine Vet J 2012;44: 143–148.
    doi: 10.1111/j.2042-3306.2011.00387.xpubmed: 21696429google scholar: lookup
  3. Theoret CL, Bolwell CF, Riley CB. A cross-sectional survey on wounds in horses in New Zealand.. N Z Vet J 2016;64: 90–94.
    doi: 10.1080/00480169.2015.1091396pubmed: 26357976google scholar: lookup
  4. Sole A, Bolwell CF, Dart AJ, Riley CB, Theoret CL. Descriptive survey of wounds in horses presented to Australian veterinarians.. Aust Equine Vet 2015;34: 68–74.
  5. Wilmink JM, van Herten J, van Weeren PR, Barneveld A. Retrospective study of primary intention healing and sequestrum formation in horses compared to ponies under clinical circumstances.. Equine Vet J 2002;34: 270–273.
    pubmed: 12108745
  6. Theoret CL, Wilmink JM. Aberrant wound healing in the horse: naturally occurring conditions reminiscent of those observed in man.. Wound Repair Regen 2013;21: 365–371.
    doi: 10.1111/wrr.12018pubmed: 23441750google scholar: lookup
  7. Weyrich LS, Dixit S, Farrer AG, Cooper AJ, Cooper AJ. The skin microbiome: Associations between altered microbial communities and disease.. Australas J Dermatol 2015;56: 268–274.
    doi: 10.1111/ajd.12253pubmed: 25715969google scholar: lookup
  8. Dowd SE, Sun Y, Secor PR, Rhoads DD, Wolcott BM, James GA. Survey of bacterial diversity in chronic wounds using pyrosequencing, DGGE, and full ribosome shotgun sequencing.. BMC Microbiology 2008;8: 43.
    doi: 10.1186/1471-2180-8-43pmc: PMC2289825pubmed: 18325110google scholar: lookup
  9. Mertz PM. Cutaneous biofilms: friend or foe?. Wounds 2003, 15, 129–132.
  10. Freeman K, Woods E, Welsby S, Percival SL, Cochrane CA. Biofilm evidence and the microbial diversity of horse wounds.. Can J Microbiol 2009;55: 197–202.
    doi: 10.1139/w08-115pubmed: 19295652google scholar: lookup
  11. Westgate SJ, Percival SL, Knottenbelt DC, Clegg PD, Cochrane CA. Microbiology of equine wounds and evidence of bacterial biofilms.. Veterinary Microbiology 2011;150: 152–159.
    doi: 10.1016/j.vetmic.2011.01.003pubmed: 21273008google scholar: lookup
  12. Jorgensen E, Bay L, Bjarnsholt T, Bundgaard L, Sorensen MA, Jacobsen S. The occurrence of biofilm in an equine experimental wound model of healing by secondary intention.. Veterinary Microbiology 2017;204: 90–95.
    doi: 10.1016/j.vetmic.2017.03.011pubmed: 28532812google scholar: lookup
  13. Ross AA, Muller KM, Weese JS, Neufeld JD. Comprehensive skin microbiome analysis reveals the uniqueness of human skin and evidence for phylosymbiosis within the class Mammalia.. Proc Natl Acad Sci U S A 2018;115: E5786–E5795.
    doi: 10.1073/pnas.1801302115pmc: PMC6016819pubmed: 29871947google scholar: lookup
  14. Lagier J-C, Hugon P, Khelaifia S, Fournier P-E, La Scola B, Raoult D. The rebirth of culture in microbiology through the example of culturomics to study human gut microbiota.. Clin Microbiol Rev 2015;28: 237–264.
    doi: 10.1128/CMR.00014-14pmc: PMC4284300pubmed: 25567229google scholar: lookup
  15. Fazli M, Bjarnsholt T, Kirketerp-Moller K, Jorgensen B, Andersen AS, Krogfelt KA. Nonrandom distribution of Pseudomonas aeruginosa and Staphylococcus aureus in chronic wounds.. Journal of Clinical Microbiology 2009;47: 4084–4089.
    doi: 10.1128/JCM.01395-09pmc: PMC2786634pubmed: 19812273google scholar: lookup
  16. Adams MK, Hendrickson DA, Rao S, Popelka FO, Bolte D. The bacteria isolated from the skin of 20 horses at a veterinary teaching hospital.. J Eq Vet Sci 2010;30: 687–695.
  17. Wolcott RD, Hanson JD, Rees EJ, Koenig LD, Phillips CD, Wolcott RA. Analysis of the chronic wound microbiota of 2,963 patients by 16S rDNA pyrosequencing.. Wound Repair Regen 2016;24: 163–174.
    doi: 10.1111/wrr.12370pubmed: 26463872google scholar: lookup
  18. Shu M, Wang Y, Yu J, Kuo S, Coda A, Jiang Y. Fermentation of Propionibacterium acnes, a commensal bacterium in the human skin microbiome, as skin probiotics against methicillin-resistant Staphylococcus aureus.. PLoS ONE 2013;8: e55380.
    doi: 10.1371/journal.pone.0055380pmc: PMC3566139pubmed: 23405142google scholar: lookup
  19. Wong VW, Martindale RG, Longaker MT, Gurtner GC. From germ theory to germ therapy: skin microbiota, chronic wounds, and probiotics.. Plast Reconstr Surg 2013;132: 854e–861e.
    pubmed: 24165637
  20. Wolcott RD, Cox SB, Dowd SE. Healing and healing rates of chronic wounds in the age of molecular pathogen diagnostics.. J Wound Care 2010;19: 272–8, 280–1.
    pubmed: 20616768
  21. Theoret CL, Barber SM, Moyana TN, Gordon JR. Expression of transforming growth factor beta(1), beta(3), and basic fibroblast growth factor in full-thickness skin wounds of equine limbs and thorax.. Vet Surg 2001;30: 269–277.
    pubmed: 11340559
  22. Bodaan CJ, Wise LM, Wakelin KA, Stuart GS, Real NC, Mercer AA. Short-term treatment of equine wounds with orf virus IL-10 and VEGF-E dampens inflammation and promotes repair processes without accelerating closure.. Wound Repair Regen 2016;24: 966–980.
    doi: 10.1111/wrr.12488pubmed: 27681311google scholar: lookup
  23. Theoret CL, Barber SM, Moyana TN, Gordon JR. Preliminary observations on expression of transforming growth factors beta1 and beta3 in equine full-thickness skin wounds healing normally or with exuberant granulation tissue.. Vet Surg 2002;31: 266–273.
    pubmed: 11994855
  24. Bigbie RB, Schumacher J, Swaim SF, Purohit RC, Wright JC. Effects of amnion and live yeast cell derivative on second-intention healing in horses.. Am J Vet Res 1991;52: 1376–1382.
    pubmed: 1928923
  25. Klindworth A, Pruesse E, Schweer T, Peplies J, Quast C, Horn M. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies.. Nucleic Acids Research 2013;41: e1–e1.
    doi: 10.1093/nar/gks808pmc: PMC3592464pubmed: 22933715google scholar: lookup
  26. Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB. Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities.. Applied and Environmental Microbiology 2009;75: 7537–7541.
    doi: 10.1128/AEM.01541-09pmc: PMC2786419pubmed: 19801464google scholar: lookup
  27. Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS. Metagenomic biomarker discovery and explanation.. Genome Biol 2011;12: R60.
    doi: 10.1186/gb-2011-12-6-r60pmc: PMC3218848pubmed: 21702898google scholar: lookup
  28. Human MPC. Structure, function and diversity of the healthy human microbiome.. Nature 2012;486: 207–214.
    doi: 10.1038/nature11234pmc: PMC3564958pubmed: 22699609google scholar: lookup
  29. Grice EA, Kong HH, Renaud G, Young AC, Bouffard GG, Blakesley RW. A diversity profile of the human skin microbiota.. Genome Research 2008;18: 1043–1050.
    doi: 10.1101/gr.075549.107pmc: PMC2493393pubmed: 18502944google scholar: lookup
  30. Alekseyenko AV, Perez-Perez GI, De Souza A, Strober B, Gao Z, Bihan M. Community differentiation of the cutaneous microbiota in psoriasis.. Microbiome 2013;1: 31.
    doi: 10.1186/2049-2618-1-31pmc: PMC4177411pubmed: 24451201google scholar: lookup
  31. Kozich JJ, Westcott SL, Baxter NT, Highlander SK, Schloss PD. Development of a Dual-Index Sequencing Strategy and Curation Pipeline for Analyzing Amplicon Sequence Data on the MiSeq Illumina Sequencing Platform.. Applied and Environmental Microbiology 2013;79: 5112–5120.
    doi: 10.1128/AEM.01043-13pmc: PMC3753973pubmed: 23793624google scholar: lookup
  32. Costa MC, Bessegatto JA, Alfieri AA, Weese JS, Filho JAB, Oba A. Different antibiotic growth promoters induce specific changes in the cecal microbiota membership of broiler chicken.. PLoS ONE 2017;12: e0171642.
    doi: 10.1371/journal.pone.0171642pmc: PMC5319738pubmed: 28222110google scholar: lookup
  33. Bessegatto JA, Paulino LR, Lisboa JAN, Alfieri AA, Montemor CH, Medeiros LP. Changes in the fecal microbiota of beef cattle caused by change in management and the use of virginiamycin as a growth promoter.. Res Vet Sci 2017;114: 355–362.
    doi: 10.1016/j.rvsc.2017.06.011pubmed: 28675873google scholar: lookup
  34. Hernandez-Chirlaque C, Aranda CJ, Ocon B, Capitan-Canadas F, Ortega-Gonzalez M, Carrero JJ. Germ-free and Antibiotic-treated Mice are Highly Susceptible to Epithelial Injury in DSS Colitis.. J Crohns Colitis 2016;10: 1324–1335.
    doi: 10.1093/ecco-jcc/jjw096pubmed: 27117829google scholar: lookup
  35. Christensen GJM, Bruggemann H. Bacterial skin commensals and their role as host guardians.. Benef Microbes 2014;5: 201–215.
    doi: 10.3920/BM2012.0062pubmed: 24322878google scholar: lookup
  36. Volz T, Skabytska Y, Guenova E, Chen K-M, Frick J-S, Kirschning CJ. Nonpathogenic bacteria alleviating atopic dermatitis inflammation induce IL-10-producing dendritic cells and regulatory Tr1 cells.. J Invest Dermatol 2014;134: 96–104.
    doi: 10.1038/jid.2013.291pubmed: 23812300google scholar: lookup
  37. Kawai T, Akira S. Toll-like receptors and their crosstalk with other innate receptors in infection and immunity.. Immunity 2011;34: 637–650.
    doi: 10.1016/j.immuni.2011.05.006pubmed: 21616434google scholar: lookup
  38. Costello EK, Lauber CL, Hamady M, Fierer N, Gordon JI, Knight R. Bacterial community variation in human body habitats across space and time.. Science 2009;326: 1694–1697.
    doi: 10.1126/science.1177486pmc: PMC3602444pubmed: 19892944google scholar: lookup
  39. Costa MC, Arroyo LG, Allen-Vercoe E, Stampfli HR, Kim PT, Sturgeon A. Comparison of the fecal microbiota of healthy horses and horses with colitis by high throughput sequencing of the V3-V5 region of the 16S rRNA gene.. PLoS ONE 2012;7: e41484.
    doi: 10.1371/journal.pone.0041484pmc: PMC3409227pubmed: 22859989google scholar: lookup
  40. Turnbaugh PJ, Hamady M, Yatsunenko T, Cantarel BL, Duncan A, Ley RE. A core gut microbiome in obese and lean twins.. Nature 2009;457: 480–484.
    doi: 10.1038/nature07540pmc: PMC2677729pubmed: 19043404google scholar: lookup
  41. Siddiqui AR, Bernstein JM. Chronic wound infection: facts and controversies.. Clin Dermatol 2010;28: 519–526.
    doi: 10.1016/j.clindermatol.2010.03.009pubmed: 20797512google scholar: lookup
  42. Han YW, Shi W, Huang GT, Kinder Haake S, Park NH, Kuramitsu H. Interactions between periodontal bacteria and human oral epithelial cells: Fusobacterium nucleatum adheres to and invades epithelial cells.. Infection and Immunity 2000;68: 3140–3146.
    pmc: PMC97547pubmed: 10816455

Citations

This article has been cited 17 times.
  1. Partusch L, Rutland CS, Martens A, Du Cheyne C, De Spiegelaere W, Michler JK. Collagen composition in equine exuberant granulation tissue reflects tissue immaturity. PLoS One 2025;20(11):e0335179.
    doi: 10.1371/journal.pone.0335179pubmed: 41196884google scholar: lookup
  2. Wesołowska M, Szczuka E. Analysis of Staphylococcal Diversity in the Skin Microbiota of Healthy Riding Horses. Antibiotics (Basel) 2025 Oct 16;14(10).
    doi: 10.3390/antibiotics14101037pubmed: 41148728google scholar: lookup
  3. Matinpour M, Zettner N, Neumann K, Bäumer L, Burkovski A. Analysis of the Culturable Skin Microbiome of Horses from Southern Germany. Microorganisms 2025 Mar 8;13(3).
    doi: 10.3390/microorganisms13030623pubmed: 40142516google scholar: lookup
  4. Allano M, Arsenault J, Archambault M, Fairbrother JH, Sauvé F. Prevalence and Risk Factors of Staphylococcus aureus Nasal Colonization in Horses Admitted to a Veterinary Teaching Hospital. J Vet Intern Med 2025 May-Jun;39(3):e70027.
    doi: 10.1111/jvim.70027pubmed: 40135807google scholar: lookup
  5. Li Y, Zhu Y, Yang B, Yu S, Li S, Wright AG, Du R, Si H, Li Z. Characteristics and Differences in the Antler Velvet Microbiota During Regeneration. Microorganisms 2024 Dec 27;13(1).
    doi: 10.3390/microorganisms13010036pubmed: 39858803google scholar: lookup
  6. Styková E, Valocký I, Kačírová J, Fecskeová LK. Microbiological effect of topically applied Weissella cibaria on equine pastern dermatitis. Front Vet Sci 2024;11:1493756.
    doi: 10.3389/fvets.2024.1493756pubmed: 39834920google scholar: lookup
  7. Khan IM, Nassar N, Chang H, Khan S, Cheng M, Wang Z, Xiang X. The microbiota: a key regulator of health, productivity, and reproductive success in mammals. Front Microbiol 2024;15:1480811.
    doi: 10.3389/fmicb.2024.1480811pubmed: 39633815google scholar: lookup
  8. Gerlicz W, Sypka M, Jodłowska I, Białkowska AM. Isolation, Selection, and Identification of Keratinolytic Bacteria for Green Management of Keratin Waste. Molecules 2024 Jul 18;29(14).
    doi: 10.3390/molecules29143380pubmed: 39064958google scholar: lookup
  9. Labens R, Raidal S, Borgen-Nielsen C, Pyecroft S, Pant SD, De Ridder T. Wound healing of experimental equine skin wounds and concurrent microbiota in wound dressings following topical propylene glycol gel treatment. Front Vet Sci 2023;10:1294021.
    doi: 10.3389/fvets.2023.1294021pubmed: 38155761google scholar: lookup
  10. Brock AK, Chamoun-Emanuelli AM, Howard EA, Huntzinger KD, Lawhon SD, Bryan LK, Cosgriff-Hernandez EM, Cohen ND, Whitfield-Cargile CM. Wound swabs versus biopsies to detect methicillin resistant Staphylococcus aureus in experimental equine wounds. Vet Surg 2022 Nov;51(8):1196-1205.
    doi: 10.1111/vsu.13872pubmed: 36102600google scholar: lookup
  11. Kaiser-Thom S, Gerber V, Collaud A, Hurni J, Perreten V. Prevalence and WGS-based characteristics of Staphylococcus aureus in the nasal mucosa and pastern of horses with equine pastern dermatitis. BMC Vet Res 2022 Feb 24;18(1):79.
    doi: 10.1186/s12917-021-03053-ypubmed: 35209904google scholar: lookup
  12. Kaiser-Thom S, Hilty M, Ramseyer A, Epper P, Gerber V. The relationship between equine pastern dermatitis, meteorological factors, and the skin microbiota. Vet Dermatol 2022 Apr;33(2):165-e48.
    doi: 10.1111/vde.13045pubmed: 34888974google scholar: lookup
  13. Sauvé F. Staphylococcal cutaneous infection in horses: From the early 2000s to the present. Can Vet J 2021 Sep;62(9):1001-1006.
    pubmed: 34475588
  14. Sangiorgio DB, Hilty M, Kaiser-Thom S, Epper PG, Ramseyer AA, Overesch G, Gerber VM. The influence of clinical severity and topical antimicrobial treatment on bacteriological culture and the microbiota of equine pastern dermatitis. Vet Dermatol 2021 Apr;32(2):173-e41.
    doi: 10.1111/vde.12912pubmed: 33417744google scholar: lookup
  15. Ruiz-López MJ. Mosquito Behavior and Vertebrate Microbiota Interaction: Implications for Pathogen Transmission. Front Microbiol 2020;11:573371.
    doi: 10.3389/fmicb.2020.573371pubmed: 33362732google scholar: lookup
  16. Saleh K, Strömdahl AC, Riesbeck K, Schmidtchen A. Inflammation Biomarkers and Correlation to Wound Status After Full-Thickness Skin Grafting. Front Med (Lausanne) 2019;6:159.
    doi: 10.3389/fmed.2019.00159pubmed: 31355202google scholar: lookup
  17. Ross AA, Rodrigues Hoffmann A, Neufeld JD. The skin microbiome of vertebrates. Microbiome 2019 May 23;7(1):79.
    doi: 10.1186/s40168-019-0694-6pubmed: 31122279google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.