title: Microbiology of equine wounds and evidence of bacterial biofilms.
abstract: Horse wounds have a high risk of becoming infected due to their environment. Infected wounds harbour diverse populations of microorganisms, however in some cases these microorganisms can be difficult to identify and fail to respond to antibiotic treatment, resulting in chronic non-healing wounds. In human wounds this has been attributed to the ability of bacteria to survive in a biofilm phenotypic state. Biofilms are known to delay wound healing, principally due to their recalcitrance towards antimicrobial therapies and components of the innate immune response. This study describes the presence of bacterial biofilms within equine wounds. Thirteen 8-mm diameter tissue samples were collected from (n=18) chronic wounds. Following histological staining, samples were observed for evidence of biofilms. Fifty one wounds and control skin sites were sampled using sterile swabs. Control skin sites were on the uninjured side of the horse at the same anatomical location as the wound. The isolated bacteria were cultured aerobically and anaerobically. The biofilm forming potential of all the isolated bacteria was determined using a standard crystal violet microtitre plate assay. Stained tissue samples provided evidence of biofilms within 61.5% (8 out of 13) equine wounds. In total 340 bacterial isolates were identified from all the equine wound and skin samples. Pseudomonas aeruginosa and Enterococcus faecium were the most predominantly isolated bacterial species from equine wound and skin samples respectively. Staphylococcus was the most commonly isolated genus in both environments. Bacteria cultured from chronic and acute wounds showed significantly (P<0.05) higher biofilm forming potential than bacteria isolated from skin. This paper highlights preliminary evidence supporting the presence of biofilms and a high microbial diversity in equine chronic wounds. The presence of biofilms in equine wounds partly explains the reluctance of many lower limb wounds to heal. Non-healing limb wounds in horses are a well documented welfare and economic concern. This knowledge can be used to shape future treatments in order to increase the healing rate and decrease the costs and suffering associate with equine wounds.
authors.affiliation: University of Liverpool, School of Veterinary Sciences, Faculty of Health and Life Sciences, Leahurst, Neston, South Wirral CH64 7TE, United Kingdom. s.j.westgate@liv.ac.uk
authors.forename: S J
authors.initials: SJ
authors.lastname: Westgate
authors.forename: S L
authors.initials: SL
authors.lastname: Percival
authors.forename: D C
authors.initials: DC
authors.lastname: Knottenbelt
authors.forename: P D
authors.initials: PD
authors.lastname: Clegg
authors.forename: C A
authors.initials: CA
authors.lastname: Cochrane
mesh_terms: D000818:Animals; D018441:Biofilms; D016984:Enterococcus faecium / Q000302:isolation & purification / Q000472:pathogenicity; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology; D006734:Horse Diseases / Q000382:microbiology*; D006736:Horses / Q000382:microbiology*; D011552:Pseudomonas Infections / Q000382:microbiology / Q000473:pathology; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification / Q000472:pathogenicity; D017192:Skin Diseases, Bacterial / Q000382:microbiology; D013203:Staphylococcal Infections / Q000382:microbiology; D013210:Staphylococcus / Q000302:isolation & purification / Q000472:pathogenicity; D014946:Wound Infection / Q000382:microbiology*