Additional file 1: of Infectious diseases during the European Union training mission Mali (EUTM MLI) – a four-year experience

Figure S1. Most commonly reported reasons for new medical consultations during the study interval for non-EUTM missions. Figure S2. Variation in the crude rate of reported disease categories year by year for non-EUTM missions. Figure S3. Variation in reported infectious disease events by category and year for non-EUTM missions. Figure S4. Variation in reported specific infectious diseases events by calendar week for non-EUTM missions. Figure S5. Reports of confirmed diseases in EUTM MLI during the study period. (DOC 2979 kb

Relative recovery of bacteria compared to inoculation dose after elution into Amies medium.

<p>Ratios were determined by comparison of viable counts of released bacteria to the inoculation dose, which was defined as 100%. Results from statistical analysis are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111627#pone.0111627.s003" target="_blank">Table S2</a>.</p

Nasal Screening for MRSA: Different Swabs – Different Results!

<div><p>Objectives</p><p>Swab-based nasal screening is commonly used to identify asymptomatic carriage of <i>Staphylococcus aureus</i> in patients. Bacterial detection depends on the uptake and release capacities of the swabs and on the swabbing technique itself. This study investigates the performance of different swab-types in nasal MRSA-screening by utilizing a unique artificial nose model to provide realistic and standardized screening conditions.</p><p>Methods</p><p>An anatomically correct artificial nose model was inoculated with a numerically defined mixture of MRSA and <i>Staphylococcus epidermidis</i> bacteria at quantities of 4×10² and 8×10² colony forming units (CFU), respectively. Five swab-types were tested following a strict protocol. Bacterial recovery was measured for direct plating and after elution into Amies medium by standard viable count techniques.</p><p>Results</p><p>Mean recovered bacteria quantities varied between 209 and 0 CFU for MRSA, and 365 and 0 CFU for <i>S. epidermidis</i>, resulting swab-type-dependent MRSA-screening-sensitivities ranged between 0 and 100%. Swabs with nylon flocked tips or cellular foam tips performed significantly better compared to conventional rayon swabs referring to the recovered bacterial yield (p<0.001). Best results were obtained by using a flocked swab in combination with Amies preservation medium. Within the range of the utilized bacterial concentrations, recovery ratios for the particular swab-types were independent of the bacterial species.</p><p>Conclusions</p><p>This study combines a realistic model of a human nose with standardized laboratory conditions to analyze swab-performance in MRSA-screening situations. Therefore, influences by inter-individual anatomical differences as well as diverse colonization densities in patients could be excluded. Recovery rates vary significantly between different swab-types. The choice of the swab has a great impact on the laboratory result. In fact, the swab-type contributes significantly to true positive or false negative detection of nasal MRSA carriage. These findings should be considered when screening a patient.</p></div

Recovery of bacteria in absolute numbers after elution into Amies medium.

<p>Viable counts of bacteria after elution of the displayed swab-types into Amies medium were determined by CFU counting as described in the methods section. CFU = colony forming units. Results from statistical analysis are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111627#pone.0111627.s002" target="_blank">Table S1</a>.</p

Recovery of bacteria in absolute numbers after direct plating.

<p>Viable counts of bacteria by direct plating of the displayed swab-types were determined by CFU counting as described in the methods section. CFU = colony forming units. Results from statistical analysis are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111627#pone.0111627.s002" target="_blank">Table S1</a>.</p

Relative recovery of bacteria compared to inoculation dose after direct plating.

<p>Ratios were determined by comparison of viable counts of released bacteria to the inoculation dose, which was defined as 100%. Results from statistical analysis are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111627#pone.0111627.s003" target="_blank">Table S2</a>.</p

Qualitative MRSA detection.

<p>Legend: Artificial nose models were inoculated with bacterial suspensions of MRSA and <i>S. epidermidis</i> and defined as 100% MRSA positive. After swabbing with five swab-types (<i>n</i> = 15 swabs per type) MRSA detection rates were analyzed by direct plating or after elution of swab contents into Amies medium. Detection of >1 CFU MRSA on Columbia agar plates supplemented with 5% sheep blood was rated as a positive result. Results of sensitivity analysis are displayed as followed: number of positive results/number of maximal positive results (percentage).</p><p>Qualitative MRSA detection.</p

Formic acid extraction triplicate spectra from the six <i>H. parainfluenzae</i> study isolates with the highest (top) and lowest (bottom) MALDI Biotyper identification scores.

<p>Inscribed numbers denote the highest score from triplicate measurements. Scores ≥2.0 constitute species-level identification. Lower-scoring spectra show reduced signal intensity and signal-to-noise ratio, resulting in reduced peak detection rates. For non-<i>H. haemolyticus</i> study isolates, failed species-level identification with the Bruker mass spectrometry fingerprinting system could be attributed to suboptimal spectrum quality rather than insufficient database coverage.</p

Rapid Discrimination of <i>Haemophilus influenzae</i>, <i>H. parainfluenzae</i>, and <i>H. haemolyticus</i> by Fluorescence <i>In Situ</i> Hybridization (FISH) and Two Matrix-Assisted Laser-Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) Platforms

<div><p>Background</p><p>Due to considerable differences in pathogenicity, <i>Haemophilus influenzae</i>, <i>H. parainfluenzae</i> and <i>H. haemolyticus</i> have to be reliably discriminated in routine diagnostics. Retrospective analyses suggest frequent misidentifications of commensal <i>H. haemolyticus</i> as <i>H. influenzae</i>. In a multi-center approach, we assessed the suitability of fluorescence <i>in situ</i> hybridization (FISH) and matrix-assisted laser-desorption-ionization time-of-flight mass-spectrometry (MALDI-TOF-MS) for the identification of <i>H. influenzae</i>, <i>H. parainfluenzae</i> and <i>H. haemolyticus</i> to species level.</p><p>Methodology</p><p>A strain collection of 84 <i>Haemophilus</i> spp. comprising 50 <i>H. influenzae</i>, 25 <i>H. parainfluenzae</i>, 7 <i>H. haemolyticus</i>, and 2 <i>H. parahaemolyticus</i> including 77 clinical isolates was analyzed by FISH with newly designed DNA probes, and two different MALDI-TOF-MS systems (Bruker, Shimadzu) with and without prior formic acid extraction.</p><p>Principal Findings</p><p>Among the 84 <i>Haemophilus</i> strains analyzed, FISH led to 71 correct results (85%), 13 uninterpretable results (15%), and no misidentifications. Shimadzu MALDI-TOF-MS resulted in 59 correct identifications (70%), 19 uninterpretable results (23%), and 6 misidentifications (7%), using colony material applied directly. Bruker MALDI-TOF-MS with prior formic acid extraction led to 74 correct results (88%), 4 uninterpretable results (5%) and 6 misidentifications (7%). The Bruker MALDI-TOF-MS misidentifications could be resolved by the addition of a suitable <i>H. haemolyticus</i> reference spectrum to the system's database. In conclusion, no analyzed diagnostic procedure was free of errors. Diagnostic results have to be interpreted carefully and alternative tests should be applied in case of ambiguous test results on isolates from seriously ill patients.</p></div

Detection of third-generation cephalosporin-resistant Enterobacteriaceae in returning soldiers.

<p>Detection of third-generation cephalosporin-resistant Enterobacteriaceae in returning soldiers.</p