<i>spa</i>-types of 91 <i>S. aureus</i> isolates from the anterior nares and wounds of 19 BU patients.

<p><i>spa</i>-types of 91 <i>S. aureus</i> isolates from the anterior nares and wounds of 19 BU patients.</p

Genetic Diversity of <i>Staphylococcus aureus</i> in Buruli Ulcer

<div><p>Background</p><p>Buruli ulcer (BU) is a necrotizing skin disease caused by Mycobacterium ulcerans. Previous studies have shown that wounds of BU patients are colonized with M. ulcerans and several other microorganisms, including <i>Staphylococcus aureus</i>, which may interfere with wound healing. The present study was therefore aimed at investigating the diversity and topography of <i>S. aureus</i> colonizing BU patients during treatment.</p><p>Methodology</p><p>We investigated the presence, diversity, and spatio-temporal distribution of <i>S. aureus</i> in 30 confirmed BU patients from Ghana during treatment. <i>S. aureus</i> was isolated from nose and wound swabs, and by replica plating of wound dressings collected bi-weekly from patients. <i>S. aureus</i> isolates were characterized by multiple-locus variable number tandem repeat fingerprinting (MLVF) and spa-typing, and antibiotic susceptibility was tested.</p><p>Principal Findings</p><p>Nineteen (63%) of the 30 BU patients tested positive for <i>S. aureus</i> at least once during the sampling period, yielding 407 <i>S. aureus</i> isolates. Detailed analysis of 91 isolates grouped these isolates into 13 MLVF clusters and 13 spa-types. Five (26%) <i>S. aureus</i>-positive BU patients carried the same <i>S. aureus</i> genotype in their anterior nares and wounds. <i>S. aureus</i> isolates from the wounds of seven (37%) patients were distributed over two different MLVF clusters. Wounds of three (16%) patients were colonized with isolates belonging to two different genotypes at the same time, and five (26%) patients were colonized with different <i>S. aureus</i> types over time. Five (17%) of the 30 included BU patients tested positive for methicillin-resistant <i>S. aureus</i> (MRSA).</p><p>Conclusion/Significance</p><p>The present study showed that the wounds of many BU patients were contaminated with <i>S. aureus</i>, and that many BU patients from the different communities carried the same <i>S. aureus</i> genotype during treatment. This calls for improved wound care and hygiene.</p></div

<i>S. aureus</i> wound topography in BU patients.

<p>Used dressings from wounds of BU patients were replica-plated onto CLED agar plates, and <i>S. aureus</i> colonies thus obtained were typed by MLVF. <i>S. aureus</i> colonies belonging to different MLVF clusters are shown in different colors: cluster A, red circles; cluster D, blue circles; cluster F, yellow circles; cluster H, green circle and cluster L, purple circles. (A) Replica plate of a wound dressing collected from patient 7 at time point t8 with <i>S. aureus</i> colonies belonging to clusters A and D, (B) Replica plate of a wound dressing collected from patient 7 at time t5 with <i>S. aureus</i> colonies belonging to clusters F and H. (C) Replica plate of a wound dressing collected from patient 22 at time t6 with <i>S. aureus</i> colonies belonging to clusters A and L.</p

Antibiotic resistances of the 91 <i>S. aureus</i> study isolates.

<p>For details on the antibiotic resistances of the respective study isolates, please see <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003421#pntd.0003421.s001" target="_blank">S1 Table</a> in the Supporting Information.</p><p>Antibiotic resistances of the 91 <i>S. aureus</i> study isolates.</p

Population structure of the 91 <i>S. aureus</i> isolates from BU patients.

<p>The population structure was obtained by BURP analysis. In the present <i>S. aureus</i> collection, comprising 13 different <i>spa</i>-types no <i>spa</i>-CCs could be generated. The BURP analysis grouped isolates together into three groups with no founder, comprising 27 (30% of all isolate), 29 (32% of all isolate) and 11 (12% of all isolate) <i>S. aureus</i> isolates, respectively. Twenty isolates (22% of all isolates) comprising six <i>spa</i>-types (46% of all <i>spa</i>-types) were identified as singletons, and four isolates (4% of all isolates) comprising one <i>spa</i>-type (t11375, 8% of all <i>spa</i>-types) were excluded. The circle size is proportional to the number of isolates in each cluster.</p

MLVF dendrogram of 91 <i>S. aureus</i> isolates from BU patients.

<p>The dendrogram was generated using the UPGMA algorithm. Additionally, seven technical controls designated M2 were included in the dendrogram. The respective MLVF clusters and <i>spa</i>-types are indicated on the right side of the dendrogram.</p

Map of Ghana indicating the region and communities from which the 19 BU patients colonized with <i>S. aureus</i> originated.

<p>Map of Ghana indicating the region and communities from which the 19 BU patients colonized with <i>S. aureus</i> originated.</p

<i>spa</i>-types of 91 <i>S. aureus</i> isolates from the anterior nares and wounds of 19 BU patients.

<p><i>spa</i>-types of 91 <i>S. aureus</i> isolates from the anterior nares and wounds of 19 BU patients.</p

Distribution of the IS<i>2404</i> target and intracellular mycobacteria from the sampling sites.

<p>Distribution of the IS<i>2404</i> target and intracellular mycobacteria from the sampling sites.</p

Isolation of FLA per type of specimen and sampling site.

<p>Isolation of FLA per type of specimen and sampling site.</p