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

ABSTRACT: OBJECTIVES: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. METHODS: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. RESULTS: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. CONCLUSIONS: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.

SUBMITTER: Kamus LJ

PROVIDER: S-EPMC6261015 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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

Kamus Louis J LJ Theoret Christine C Costa Marcio C MC

PloS one 20181126 11

<h4>Objectives</h4>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.<h4>Methods</h4>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 woun ...[more]

PMID: 30475922

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Project description:BackgroundBetween 2008 and 2011 about 40 cases of human cowpox were reported from Germany and France. Infections had been acquired via close contact to infected, young pet rats. An identical and unique sequence of the hemagglutinin gene was found in various cowpox virus (CPXV) isolates pointing to a common source of infection. In a second CPXV outbreak in cats in a small animal clinic in Germany in 2015, four out of five hospitalized cats showed identical hemagglutinin sequences and thus, a hospital-acquired transmission had been assumed. Next-Generation Sequencing was performed in order to re-investigate the outbreaks, as epidemiological data could not confirm all cases.MethodsHomogenates of lesion material from rats, cats and humans were cultivated in cell culture. The genomes of four virus isolates, nine CPXVs from our strain collections and from DNA of three paraffin-embedded lesion materials were determined by Next Generation Sequencing (NGS). For phylogenetic analyses a MAFFT-alignment was generated. A distance matrix based on concatenated SNPs was calculated and plotted as dendrogram using Unweighted Pair Group Method with Arithmetic mean (UPGMA) for visualization.ResultsAligning of about 200.000 nucleotides of 8 virus isolates associated with the pet rat outbreak revealed complete identity of six genomes, the remainder two genomes differed in as little as 3 SNPs. When comparing this dataset with four already published CPXV genomes also associated with the pet rat outbreak, again a maximum difference of 3 SNPs was found. The outbreak which lasted from 2008 till 2011 was indeed caused by a single strain which has maintained an extremely high level of clonality over 4 years. Aligning genomic sequences from four cases of feline cowpox revealed 3 identical sequences and one sequence which differed in 65 nucleotides. Although identical hemagglutinin sequences had been obtained from four hospitalized cats, genomic sequencing proved that a hospital-acquired transmission had occurred in only three cats.ConclusionAnalyzing the rather short sequence of the hemagglutinin gene is not sufficient to conduct molecular trace back analyses. Instead, whole genome sequencing is the method of choice which can even be applied to paraffin-embedded specimens.

Dataset Information

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

Publications

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

Similar Datasets

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