Specific antibacterial activity of copper alloy touch surfaces in five long-term care facilities for older adults.

BACKGROUND: Pathogens involved in healthcare-associated infections can quickly spread in the environment, particularly to frequently touched surfaces, which can be reservoirs for pathogens. AIM: The purpose of this study was to investigate naturally occurring bacterial contamination on touch surfaces in five French long-term care facilities and to compare bacterial populations recovered from copper and control surfaces. METHODS: More than 1300 surfaces were sampled. The collected bacteria were identified to obtain a global view of the cultivable bacterial populations colonizing touch surfaces. Haemolytic colonies and putative pathogens were also screened using specific agar plates and then identified with matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. In total, more than 3400 colonies were analysed. FINDINGS: Staphylococcus and Micrococcus were the two predominant genera present on touch surfaces, respectively occurring on 51.8% and 48.0% of control surfaces. In these facilities with relatively low bioburden, copper surfaces efficiently reduced the occurrence frequencies of three genera: Staphylococcus, Streptococcus and Roseomonas. Pathogenic species such as Staphylococcus aureus, Enterococcus faecalis and E. faecium were observed in very few samples. In addition, meticillin-resistant S. aureus was observed on five control surfaces and one copper surface. CONCLUSION: Contamination of healthcare facilities touch surfaces can be the source for the spread of bacteria through the institution. This in situ study shows that the frequency of the contamination as well as the specific bacterial population bioburden is reduced on copper alloy surfaces.

Amoxicillin-tolerant Pasteurella multocida strain isolated from chronic dermohypodermitis after suboptimal exposure to amoxicillin is not associated with reduced growth rate.

Pasteurella multocida is rarely observed in human chronic infections. A Pasteurella multocida strain was isolated from a skin biopsy of chronic dermohypodermitis in a 21-year-old woman without an immunocompromised state. As this strain was viable one month after a cat scratch despite treatment by amoxicillin-clavulanic acid, we compared this strain's growth rate, amoxicillin Minimal Inhibitory and Bactericidal Concentrations (MIC and MBC), resistance to serum and ability to activate neutrophil granulocytes with those of control strains isolated during acute infections in humans without previous antibiotics exposure. This particular strain was not more resistant to serum and did not induce a lower phagocytic activity than control strains. It did not grow more slowly than control strains even after suboptimal exposure to amoxicillin. This particular strain was tolerant to amoxicillin but tolerance did not appear sufficient alone for the induction of a chronic infection in a host without an immunocompromised state.

Characterization of quinolone resistance mechanisms in Enterobacteriaceae recovered from diseased companion animals in Europe.

ComPath is a European monitoring programme dedicated to the collection of bacterial pathogens from diseased dogs and cats to determine their antibiotic susceptibility. The objective was to characterize genetic determinants associated with quinolone resistance among 69 enrofloxacin non-wild type strains selected among 604 non-duplicate Enterobacteriaceae isolates collected in 10EU countries from 2008 to 2010: quinolone resistance determining region (QRDR) and plasmid-mediated quinolone resistance (PMQR). Among them, 17% (12/69) carried at least one PMQR (9/12 qnrB, qnrS or qnrD and 4/12 aac(6')-Ib-cr) and 83% (57/69) no PMQR. All the Klebsiella pneumoniae isolates chromosomally carried oqxAB . No qepA genes were detected. Eight strains did not carry any mutations in QRDR (4 PMQR-positive and 4 PMQR-negative strains). From the 12 PMQR-positive strains, 4 showed enrofloxacin MICs≤2µg/mL, and 8 MICs≥8µg/mL (resistant). These latter strains carried 1-5 mutations in QRDR, including a ParE I529L mutation. qnrD was found in 2 Proteus mirabilis and the plasmids were similar to pDIJ09-518a previously described. For the 57 non-PMQR strains, 29 strains showed MICs≤2µg/mL (4 with no QRDR mutations, 21 with 1 mutation in GyrA, 4 with 2 mutations in GyrA) and 28 showed enrofloxacin MICs≥8µg/mL carrying at least 2 mutations in QRDR, including a ParE I529L mutation for 2 Escherichia coli strains with a total of 5 QRDR mutations. No GyrB mutations were found. qnr was the major PMQR and qnrD was only detected in Proteus spp. Twelve strains carried at least 4 mutations.

Molecular epidemiology of OXA-48-producing Klebsiella pneumoniae in France.

We characterized 53 OXA-48-producing Klebsiella pneumoniae (OXA-48-Kp) isolated between 2011 and 2013 in 21 French hospitals. All the isolates were genotyped using MLST and PFGE and the population structure of the species was determined by a nucleotide-based analysis of the entire K. pneumoniae MLST database. Most of the OXA-48-Kp isolates also produced CTX-M-15 and remained susceptible to imipenem and meropenem. The isolates were distributed into 20 STs, of which five were dominant (ST15, ST101, ST147, ST395 and ST405). All the OXA-48-Kp clustered in the major clade of K. pneumoniae KpI.

Ciprofloxacin treatment failure in a murine model of pyelonephritis due to an AAC(6')-Ib-cr-producing Escherichia coli strain susceptible to ciprofloxacin in vitro.

AAC(6')-Ib-cr is a plasmid-mediated quinolone resistance mechanism described worldwide for Escherichia coli. Since it confers in vitro only a low level of resistance to ciprofloxacin, we evaluated its impact on the in vivo activity of ciprofloxacin. Isogenic strains were obtained by transferring plasmid p449, harboring aac(6')-Ib-cr, into the quinolone-susceptible strain E. coli CFT073-RR and its D87G gyrA mutant. MICs were 0.015, 0.06, 0.25, and 0.5 µg/ml against E. coli strains CFT073-RR, CFT073-RR/p449, CFT073-RR GyrA(r), and CFT073-RR GyrA(r)/p449, respectively. Bactericidal activity was reduced at 1× the MIC for the three resistant derivatives, while at a fixed concentration of 0.5 µg/ml, 99.9% killing was observed for all strains except E. coli CFT073-RR GyrA(r)/p449. In the murine model of pyelonephritis, an optimal regimen of ciprofloxacin (10 mg/kg of body weight twice a day [b.i.d.]) significantly decreased the bacterial count in the kidneys of mice infected with E. coli CFT073 (1.6 versus 4.3 log10 CFU/g of kidney compared to untreated controls; P = 0.0001), while no significant decrease was observed for E. coli CFT073-RR/p449 (2.7 versus 3.1 log10 CFU/g; P = 0.84), E. coli CFT073-RR GyrA(r) (4.2 versus 4.1 log10 CFU/g; P = 0.35), or E. coli CFT073-RR GyrA(r)/p449 (2.9 versus 3.6 log10 CFU/g; P = 0.47). While pharmacokinetic and pharmacodynamic (PK/PD) parameters accounted for ciprofloxacin failure against gyrA-containing mutants, this was not the case for the aac(6')-Ib-cr-containing strains, suggesting an in situ hydrolysis of ciprofloxacin in the latter case.

Gastroscopy-associated transmission of extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa.

An unusual multi-drug-resistant Pseudomonas aeruginosa (MDR-PA) was isolated in four patients whilst hospitalized in a French teaching hospital between May and August 2011. All four patients had undergone an oesophago-gastro-duodenoscopy with the same gastroscope over a five-month period. This endoscope was associated with a culture positive for the MDR-PA. Observations of endoscope reprocessing identified deviations from the agreed processes: insufficient initial cleaning, shortening of the immersion time and brushing time, insufficient channel flushing, and inadequate drying prior to storage. Since withdrawing the gastroscope and institution of strict adherence to the agreed processes, no other MDR-PA cases have been isolated.

Raman spectroscopic analysis of the clonal and horizontal spread of CTX-M-15-producing Klebsiella pneumoniae in a neonatal intensive care unit.

Nosocomial outbreaks of extended-spectrum ß-lactamase (ESBL)-producing Klebsiella pneumoniae are an increasing concern in neonatal intensive care units (NICUs). We describe an outbreak of ESBL-producing K. pneumoniae that lasted 5 months and affected 23 neonates in our NICU. Proton pump inhibitor and extended-spectrum cephalosporin exposure were significantly associated with the risk of ESBL-producing K. pneumoniae colonisation and/or infection. Thirty isolates recovered from clinical, screening and environmental samples in the NICU were studied by means of Raman spectroscopy, pulsed-field gel electrophoresis and repetitive extragenic palindromic polymerase chain reaction (rep-PCR). The Raman clustering was in good agreement with the results of the other two molecular methods. Fourteen isolates belonged to the Raman clone 1 and 16 to the Raman clone 3. Molecular analysis showed that all the strains expressed SHV-1 chromosomal resistance, plasmid-encoded TEM-1 and CTX-M-15 ß-lactamases. Incompatibility groups of plasmid content identified by PCR-based replicon typing indicated that resistance dissemination was due to the clonal spread of K. pneumoniae and horizontal CTX-M-15 gene transfer between the two clones.

[Unusual location of a brain abscess due to Listeria monocytogenes]. / Abcès cérébral à Listeria monocytogenes de localisation inhabituelle.

Here we report a case of sustentorial brain abscess due to Listeria monocytogenes. Blood culture and procalcitonine blood measurement were negative. L. monocytogenes was isolated from CSF after inoculation in Castañeda medium.

[Predictive factors of acquisition of epidemic extended-spectrum beta-lactamase-producing Escherichia coli]. / Etude des facteurs prédictifs d'acquisition d'une souche épidémique d'Escherichia coli productrice de bêta-lactamase à spectre étendu.

The aim of the study was to determine predictive factors of epidemic extended spectrum beta-lactamase-producing Escherichia coli acquisition. All patients presenting any type of culture positive for ESBL-producing E. coli between November 2006 and October 2007 were included. An epidemic case was defined as a patient colonized with a clonal strain having epidemiological link with an another patient harbouring the same strain. Clinical and administrative data were recorded. Groups were compared by univariate and multivariate analysis using SAS software. Were included in this study 148 patients: 60 epidemic and 88 sporadic cases. Multivariate analysis showed several predictive factors of epidemic strain acquisition: female gender, high Charlson index, treatment by amoxicillin or ticarcillin-clavulanic acid, admission to emergency unit and hospitalisation in a high number of different care units. Identification of predictive factors of epidemic ESBL-producing E. coli strains acquisition may help to limit cross transmission of such strains.

[Real-time PCR for fast detection of plasmid-mediated qnr genes in extended spectrum beta-lactamase producing Enterobacteriaceae]. / Mise au point d'une technique de PCR en temps réel pour la détection rapide des gènes qnr chez des entérobactéries productrices de bêta-lactamases à spectre étendu.

AIM OF THE STUDY: To develop a fast and reliable real time PCR technique for detecting plasmid-mediated quinolone resistance genes qnrA, qnrB and qnrS. METHODS: A real-time PCR assay using SYBR Green I and Roche LightCycler(®) was developed to detect qnr genes. Detection of qnr genes was based on comparison of melting temperature differences with a positive control of each qnr genes. This assay was performed to study 138 isolates collected from diagnostic and screening samples in the Champagne-Ardenne region in 2004 (France). RESULTS: In optimized conditions, the three positive controls tested alone and with isolates confirmed the specificity of the PCR primers. Each PCR assay was able to test 30 strains in 60min for 1 qnr gene. Out of 138 isolates screened, 3.6 % isolates were positive for a qnrA1, 1.5 % for qnrS1 and no qnrB-like gene. Prevalence of qnr determinants was 5 % and reached 9.5 % in clinical isolates. CONCLUSION: Real-time PCR is a fast and reliable technique for screening of qnr-positive strains. This study shows a relatively high prevalence of qnr determinants (5 %) among ESBL-producing Enterobacteriaceae.

[Nocardia brain abscess: features, therapeutic strategies and outcome]. / Abcès cérébraux àNocardia: caractéristiques radiocliniques et prise en charge thérapeutique.

BACKGROUND AND PURPOSE: Nocardia species is an aerobic soil-saprophyte bacterium, responsible for rare opportunistic infections, mainly reported in immunocompromised patients. Nocardia brain abscess accounts for 1 to 2% of cerebral abscesses. Prognosis is poor. METHODS: We describe clinical, radiological and bacteriological findings along with therapeutic aspects for five patients and review the literature on Nocardia cerebral abscess. RESULTS: The clinical features of Nocardia brain abscess are insidious and nonspecific, occurring frequently with a medical background of obvious or latent immunodeficiency; fever, if any, is observed subordinate to extracerebral nocardiosis. Computerized tomography scan and conventional magnetic resonance (MR) scan show lesions with a necrotic core and multilobed thick walls enhancing after injection of gadolinium or iodine. Abscesses are mainly located in the brain stem, basal ganglia and cerebral cortex of the frontal, parietal and occipital lobes; cerebellar and spinal locations are uncommon. MR diffusion-weighted imaging with calculation of apparent diffusion coefficient and proton MR spectroscopy can provide additional data for accurate differential diagnosis between abscess and other necrotic lesions, such as tumor and cyst formations. Bacteriological identification has progressed with advances in molecular microbiology: 16S rRNA sequencing, allowing a more rapid routine identification of Nocardia strains from clinical samples. Clinical management of patients with a Nocardia brain abscess relies upon early use of intravenous antibiotics adapted to the strains identified and their susceptibility. Most Nocardia strains display susceptibility to cotrimoxazol, amikacin and linezolid, but develop beta-lactamase activity. CONCLUSIONS: Early pus samples, obtained by biopsy or surgical resection, are needed to establish a certain bacteriological diagnosis and initiate appropriate intravenous antibiotics.

Leptospira exposure in the human environment in France: A survey in feral rodents and in fresh water.

This paper confirms the important role of rodents to be maintenance hosts of leptospires. Their role is related to renal carriage and shedding of leptospires into urine, thus contaminating fresh water. Serological and carriage of feral rodents trapped in France were determined by MAT and hap1PCR specific for pathogenic leptospires. In same areas, fresh water samples were analyzed by hap1PCR. The overall seroprevalence was 44% in 649 rodents and was similar regardless of the species. Seroprevalence for leptospirosis is about 20-53% according to species. hap1PCR (516 kidneys) showed that renal carriage was higher in brown rats (34.7%) and muskrats (15.8%) than in coypus (3.3%). hap1PCR demonstrates a significative difference (P-value > 10(-12)) for the renal carriage between the different species: muskrats and rats are more efficient maintenance hosts than coypu but all infect water. Moreover 5/38 water samples associated with human cases were hap1PCR positive and 1/113 in controlled waters.

[Epidemiological and clinical study of Mycoplasma pneumoniae respiratory infections in children hospitalized in a pediatric ward between 1999 and 2005 at the Reims University Hospital]. / Etude épidémiologique et clinique des infections respiratoires àMycoplasma pneumoniae chez les enfants hospitalisés en pédiatrie entre 1999 et 2005 au CHU de Reims.

OBJECT: To determine the frequency, clinical features, and morbidity of Mycoplasma pneumoniae infections. METHOD: Retrospective study of 76 consecutive children under 16 years of age hospitalized at the Reims University Hospital from 1999 to 2005 with M. pneumoniae pneumonia. The infection was defined by the presence of M antibodies and/or an increase in G antibodies (quantitative Elisa test). RESULTS: M. pneumoniae was the cause of 16% (76/464) of hospitalized pneumonia cases. A significantly increased frequency was observed in 2004 (34%; 19/56) and 2005 (26%; 22/84) versus 11% from 1999 to 2003, p<5.10(-4). The mean age of the patients was 6 years and 8 months, with a peak at 3 years (14/76; 18% of patients). The most frequent clinical feature was cough (80%; 56/70). The chest X-ray showed typical radiological features such as peribronchial and perivascular interstitial infiltrates in only 23% (16/69). Respiratory and extrarespiratory complications were seen in 17 and 12 children, respectively. Only 1 child suffered from respiratory sequelae. CONCLUSION: M. pneumoniae pneumonia is frequent in children over 2 years of age. The diagnosis is sometimes difficult to initially assert because there are no specific features. Respiratory and extrarespiratory complications remain possible. Respiratory sequelae can still exist even if most cases evolve favorably under treatment by macrolides.

Incidence of class A extended-spectrum beta-lactamases in Champagne-Ardenne (France): a 1 year prospective study.

OBJECTIVES: To assess the frequency and diversity of extended spectrum beta-lactamases (ESBLs) in the Champagne-Ardenne region France, and to identify genetic elements associated with the bla(CTX-M) genes. METHODS: During 2004, all the non-duplicate isolates of Pseudomonas aeruginosa and Acinetobacter baumannii resistant to ceftazidime and of Enterobacteriaceae intermediate or resistant to ceftazidime and/or cefotaxime, screening samples excluded, were collected in 10 public hospitals and 3 private clinics. bla genes were sequenced and bla(CTX-M) environment characterized by PCR mapping. RESULTS: In Enterobacteriaceae (138/21 861; 0.6%), ESBLs were predominantly TEM-24 (n = 52; 37.7%) and CTX-M-15 (n = 37; 26.8%). Three new enzymes were identified, CTX-M-61 (CTX-M-1 group), TEM- and SHV-type. A. baumannii (n = 5) produced VEB-1 and P. aeruginosa (n = 2) SHV-2a. ISEcp1 was detected in 22/27 strains, disrupted in 7 of them. The IS903-like element was downstream of bla(CTX-M-14) and bla(CTX-M-16). ISCR1 was found upstream of bla(CTX-M-2) and bla(CTX-M-9), and ISCR1 and bla(CTX-M-2) were located on a sul1-type class 1 integron. In comparison with 2001-02, ESBL distribution among Enterobacteriaceae showed an increase in CTX-M-type (44.9% vs 3.7% P < 10(-7)) due to Escherichia coli CTX-M-15 and to the almost total disappearance of TEM-3 (0.9% vs 51.2%). E. coli was the most frequent species (50.0% vs 5.1% in 1998) despite a similar prevalence to that in 1998 (0.5% vs 0.2%). CONCLUSIONS: A careful detection of bla(CTX-M)-type spread to other species would help to anticipate clonal endemics such as those observed in Enterobacter aerogenes TEM-24.

FT-IR microspectroscopy for early identification of some clinically relevant pathogens.

AIMS: To investigate the potentials and limitations of Fourier transform-infrared (FT-IR) microspectroscopy as a tool to identify, at the level of microcolonies, pathogenic bacteria frequently isolated in the clinical environment. METHODS AND RESULTS: A total of 1570 FT-IR spectra from 164 gram-positive and gram-negative bacteria isolated from patients were recorded from 6 to 10-h old microcolonies of 50-150 microm size. A classification of 100% was obtained for the most frequent gram-positive bacteria, such as Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and Enterococcus faecium at the species level. An average accuracy of about 80% was reached with Gram negative bacteria from the Enterobacteriaceae and Pseudomonaceae families; Enterobacter aerogenes, Enterobacter cloacae, Klebsiella spp., and Citrobacter koseri; and Proteus mirabilis and Escherichia coli. Results were comparable with FT-IR measurements on dried suspensions from 18-h cultures. CONCLUSIONS: Early identification of young microcolonies is feasible with FT-IR microscopy with a very high accuracy for gram-positive bacteria. Some improvement in the transfer of microcolonies is necessary to increase the accuracy for gram-negative bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Combination of FT-IR microscopy and multivariate data analysis could be a complementary, rapid, and reliable tool for screening and discriminating, at species and subspecies level, micro-organisms of clinical, food-borne, or environmental origins.

[Legionellosis due to Legionella gormanii fortuitously found in a man with chronic lymphocytic leukemia: a case study and literature review]. / Légionellose à Legionella gormanii de découverte fortuite et leucémie lymphoïde chronique: à propos d'un cas et revue de la littérature.

Legionellosis due to other species than Legionella pneumophila is rarely described in human cases. It has been reported in immunocompromised patients with respiratory symptoms of pneumonia. We report a case of legionellosis in an immunocompromised 54-year-old man hospitalized for a blood transfusion. A routine pulmonary X- Ray was made and then a bronchoalveolar lavage was collected in which Legionella gormanii was identified. The diagnostic of legionellosis must be considered in all immunocompromised patients presenting with any pulmonary symptoms.