Diversity of <em>Acinetobacter baumannii</em> in Four French Military Hospitals, as Assessed by Multiple Locus Variable Number of Tandem Repeats Analysis

<div><h3>Background</h3><p>Infections by <em>A. calcoaceticus</em>-<em>A. baumannii</em> (ACB) complex isolates represent a serious threat for wounded and burn patients. Three international multidrug-resistant (MDR) clones (EU clone I-III) are responsible for a large proportion of nosocomial infections with <em>A. baumannii</em> but other emerging strains with high epidemic potential also occur.</p> <h3>Methodology/Principal Findings</h3><p>We automatized a Multiple locus variable number of tandem repeats (VNTR) analysis (MLVA) protocol and used it to investigate the genetic diversity of 136 ACB isolates from four military hospitals and one childrens hospital. <em>Acinetobacter</em> sp other than <em>baumannii</em> isolates represented 22.6% (31/137) with a majority being <em>A. pittii</em>. The genotyping protocol designed for <em>A.baumannii</em> was also efficient to cluster <em>A. pittii</em> isolates. Fifty-five percent of <em>A. baumannii</em> isolates belonged to the two international clones I and II, and we identified new clones which members were found in the different hospitals. Analysis of two CRISPR-cas systems helped define two clonal complexes and provided phylogenetic information to help trace back their emergence.</p> <h3>Conclusions/Significance</h3><p>The increasing occurrence of <em>A. baumannii</em> infections in the hospital calls for measures to rapidly characterize the isolates and identify emerging clones. The automatized MLVA protocol can be the instrument for such surveys. In addition, the investigation of CRISPR/cas systems may give important keys to understand the evolution of some highly successful clonal complexes.</p> </div

Polymorphism of the AYE CRISPR locus in <i>A. baumannii</i>.

<p>A) Schematic representation of the CRISPR locus in three sequenced genomes and at the growing end of isolates A28, P054 and P065. Arrows show the position of primers used to amplify a portion of the locus. Specific spacers are shown with grey boxes. B) Sequence of the growing end of strain AB307 and isolate A28. The sequence flanking the last DR is in italics.</p

Genetic diversity of 56 <i>A. baumannii</i> isolates.

<p>The legend is that of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044597#pone-0044597-g001" target="_blank">Figure 1</a>.</p

Oligonucleotides used for the multiplex PCR reactions.

a<p>The forward primer is labeled with one of three dyes D2, D3 and D4. The allele range for each reaction is indicated showing that amplicons with the same dye cannot overlap.</p>b<p>Values referred to the genome of strain ATCC 17978 and considered for allele assignment.</p>c<p>The allele range for each reaction is indicated showing that amplicons with the same dye cannot overlap. Values obtained with <i>A. baumannii</i> isolates.</p

Occupational Exposures to Ebola Virus in Ebola Treatment Center, Conakry, Guinea

We report 77 cases of occupational exposures for 57 healthcare workers at the Ebola Treatment Center in Conakry, Guinea, during the Ebola virus disease outbreak in 2014−2015. Despite the high incidence of 3.5 occupational exposures/healthcare worker/year, only 18% of workers were at high risk for transmission, and no infections occurred