[Staphylococcus saprophyticus isolated from urine culture in outpatients: epidemiology and antimicrobial susceptibility (Label Bio Elbeuf study - November 2007-July 2009)]. / Staphylococcus saprophyticus isolés d'examens cytobactériologiques urinaires en ville : épidémiologie et sensibilité aux antibiotiques (étude Label Bio Elbeuf - novembre 2007-juillet 2009).

OBJECTIVE: The aim of the study was to determine the distribution and the antibiotic susceptibility of Staphylococcus saprophyticus patterns isolated from urine culture in outpatients (population: 57,000, Elbeuf, Normandie, France). DESIGN: Prospective study from November 2007 to October 2009 in collaboration with three private medical laboratories. Determination of susceptibility to oxacillin by disk diffusion (cefoxitin, and moxalactam), automated method (Vitek BioMérieux 2) and mecA PCR's detection. Determination of the minimum inhibitory concentration by microbroth dilution for other antibiotics. RESULTS: Five thousand and fifty-one bacterial strains isolated, 91 strains of S. saprophyticus (1.8%), 89 in women (2.25%) and two in men (0.18%). S. saprophyticus represented 10.3% and 14.5% of isolates (women respectively aged between 11 and 30; 16 and 20 years); S. saprophyticus is isolated less frequently in winter. mecA PCR detection was positive for two strains. All strains tested were susceptible to ciprofloxacin and furans. Only one strain is resistant to cotrimoxazole. CONCLUSIONS: S. saprophyticus is found mostly in women between 11 to 30 years. Cotrimoxazole (after susceptibility testing) is efficient in case of S. saprophyticus's cystitis. Furans (probabilistic treatment) have to be reevaluated because of the potential for serious adverse effects.

Assessment of the bio-preparedness and of the training of the French hospital laboratories in the event of biological threat.

A national laboratory network 'Biotox-Piratox' was created in 2003 in France with the purpose of detecting, confirming and reporting potential biological and chemical threat agents. This network is divided into three levels: Level 1 is dedicated to the evaluation of risks (biological, chemical, radiological), to sampling and packing. Level 2 consists of university and military hospitals, who deal with biological specimens, and of environmental and veterinary laboratories, who deal with environmental and animal samples. Level 3 comprises national reference laboratories and the Jean Mérieux biosafety level (BSL)-4 laboratory in Lyon. This report presents the results of four bio-preparedness exercises to check critical points in the processing of samples. These exercises took place in 2007, 2009, 2010 and 2011. Each of them consisted of two parts. The first part was the identification of an unknown bacterial strain and its susceptibility to antibiotics used as a default in case of a bioterrorist event. The second part was the detection of Class III microorganisms, mainly by molecular techniques. The main lesson learnt in these exercises was that the key to successful detection of biological agents in case of a biological threat was standardisation and validation of the methods implemented by all the laboratories belonging to the network.

[Fecal carriage of third-generation cephalosporins-resistant Enterobacteriaceae in asymptomatic young adults: evolution between 1999 and 2009]. / Portage digestif d'entérobactéries résistantes aux céphalosporines de troisième génération dans une population d'adultes jeunes asymptomatiques : évolution entre 1999 et 2009.

OBJECTIVES: The aim of this work was to evaluate the fecal carriage of third generation cephalosporins resistant Enterobacteriaceae in nonhospitalized asymptomatic young adults. METHODS: A total of 517 normal fecal samples were spread onto plates agar containing cefotaxime. Isolated strains were identified and studied with agar disk diffusion antibiogram, minimal inhibition concentration in liquid medium and phenotypic and molecular study. Data were compared with a previous study realised in the same conditions in 1999. RESULTS: In 2009, the prevalence of cefotaxime resistant enterobacteria was 4.2%. Of these 22 Enterobacteriaceae, 11 harboured overexpressed cephalosporinase and 11 produced extended-spectrum-betalactamase (ESBL). Among ESBL, six E. coli produced CTX-M from group 1 (n=6), group 2 (n=1), group 9 (n=2), one E. coli produced SHV-12 and one Klebsiella pneumoniae produced CTX-M from group 1. All ESBL were multiresistant. In 1999, all the CTX resistant isolates recovered produced a cephalosporinase and no ESBL was found. CONCLUSIONS: This study highlights the increasing prevalence of fecal carriage of ESBL-producing enterobacteria in asymptomatic young patients in the community (0% in 1999 versus 2.1% in 2009; P<0.001). E. Coli with CTX-M from group 1 was the most frequent ESBL identified, while fecal carriage of Enterobacteteriaceae overproducing cephalosporinase was similar (2.1%).

[Susceptibility to antibiotics of Escherichia coli isolated from community-acquired urinary tract infections]. / Sensibilité aux antibiotiques des Escherichia coli isolés d'infections urinaires communautaires.

OBJECTIVE: The authors had for aim to determine the distribution of bacterial isolates and the antibiotic susceptibility patterns of Escherichia coli from community-acquired urinary tract infections in an area covering 57,000 people (Elbeuf, Normandie, France). DESIGN: From November 2007 to October 2008, three private French laboratories consecutively collected 2344 bacteria including 1636 E. coli from outpatients. The antibiotic susceptibility of E. coli was determined using an automatized method (Vitek 2 Biomerieux). RESULTS: The global susceptibility of E. coli was: ampicillin: 57%; amoxicillin+clavulanic acid: 73%; cefixim: 96%; ceftriaxone: 98%; gentamycin: 96%; nalidixic acid: 82%; ciprofloxacin: 89%; fosfomycin: 98%; nitrofurantoin: 96% and cotrimoxazole: 81%. The susceptibility of E. coli to ciprofloxacin was higher in 15- to 65-year-old female patients (94%) than for older female (85%) or male patients (80%). CONCLUSIONS: In the Elbeuf area, third generation cephalosporins, aminoglycosides, nitrofurantoin, and fosfomycin were the most effective on E. coli isolated from community-acquired urinary tract infections. Fluoroquinolones were more active in 15- to 65-year-old female patients than in male patients and in female patients over 65 years of age. The rates of acquired resistance were related to the level of antibiotic prescription in the various populations.

[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.

[Pseudomonas aeruginosa and beta-lactam antibiotics at the time of Europe]. / Pseudomonas aeruginosa et bêta-lactamines à l'heure de l'Europe.

Breakpoints harmonization concerning six European committees is being conducted within the European Committee for Antimicrobial Susceptibility Testing for a few years and is now finalized for beta-lactam antibiotics. This article describes the impact of breakpoint modifications on percentage of susceptibility of P. aeruginosa to beta-lactams in comparison with previous French National Committee breakpoints (CA-SFM). This harmonization leads to the need for new recommendations about diameter breakpoints and for updating breakpoints in antibiotic susceptibility testing automated devices. Moreover, it points out the importance of MICs and quantitative diameter data in order to follow the evolution of antibiotic susceptibility.

[Automation and consolidation of clinical biology: the HIA Begin experiment]. / Automatisation et consolidation en biologie clinique: l'expérience de l'hôpital d'instruction des armées Bégin.

Reorganization of hospital or private sector labs is a strong trend. Such process implies consolidation of tests on a minimum of analyzers, harmonization of equipment (limitation of number of suppliers), posts number reduction, and development of powerful software. Increase of productivity needs set up of automation systems, specially at the level of pre-analytical phase, in a way to limit manual steps. At the same moment, quality improvement generates a decreasing TAT (Turn-Around-Time), a strong reduction of errors, a better protection against biological hazards, and standardization of sample management and validation process. After giving some notions about consolidation and integration, our choices for a brand new and innovative solution are explained.

[Antibacterial spectrum of an antibiotic and clinical categorization]. / Spectre d'activité antibactérien d'un antibiotique et catégorisation clinique.

The clinical categorization of a bacterial strain for an antibiotic (SIR system) is based on the in vitro determination of minimum inhibitory concentration and confrontation with breakpoints proposed by national or international committees. The antibiotic spectrum presented in the summary of product characteristics and established by national or European agencies aims at classifying each bacterial species according to their general susceptibility level for the antimicrobial agent. Clinical categorization and antibiotic spectrum are closely related to the definition of breakpoints, established on pharmacokinetics/pharmacodynamics parameters, distribution of minimum inhibitory concentrations for relevant bacteria, acquired resistance rates and if possible, correlation with clinical outcome. This article underlines the need for standardization of susceptibility testing methods, harmonization of breakpoints between European national guidelines, and clinical trial data assessing the correlation between the minimum inhibitory concentration and therapeutic success or failure.

[Infectious aetiologies of travelers' diarrhoea]. / Les étiologies infectieuses des diarrhées du voyageur.

Traveler's diarrhoea (TD) occurs in 20 to 60% of European or North-American travelers in intertropical areas. Following return from endemic zone, malaria must always be evocated in front of febrile diarrhoea. Many causative infectious agents are involved in TD and their frequency may vary according to destination and seasons. The main agents involved in TD are Escherichia coli pathovars (especially enterotoxigenic and enteroaggregative E. coli) followed by enteroinvasive bacteria (Campylobacter spp., Shigella spp., Salmonella enterica), enteric viruses (norovirus, rotavirus) and protozoa (Gardia intestinalis, Cryptosporidium parvum et Entamoeba histolytica). The development of molecular biology methods as PCR may allow us to evaluate the relative frequency of these agents and especially of viral agents in TD. Protozoa and microsporidia are more frequently isolated in persistent and chronic TD, especially in compromised patients. A complete etiological research in routine microbiology laboratories is difficult and time-consuming, related to the high diversity of causative agents and the need for specific methods. Implementation of laboratory diagnosis is highly recommended when diarrhoea is associated with fever or presence of blood in stools, immunosuppression, antibiotic treatment (Clostridium difficile toxins) or in case of persistent/chronic diarrhoea. According to the high frequency of acquired antibiotic-resistance in enteric bacteria, an antibiogram must be performed for all causative bacterial agents.

Susceptibility of Pseudomonas aeruginosa to antimicrobials: a 2004 French multicentre hospital study.

OBJECTIVES: Pseudomonas aeruginosa is a major causative agent of hospital infections. The purpose of this study was to determine the antibiotic susceptibility of P. aeruginosa in a French multicentre study and to investigate the mechanisms of beta-lactam resistance. METHODS: Four hundred and fifty non-repetitive strains of P. aeruginosa were collected in 15 French university hospitals in 2004. MICs of antibiotics were measured by agar dilution methods. For all the strains with MICs of ticarcillin >16 mg/L, detection and identification of the beta-lactamases, quantitative determination of cephalosporinase and overproduction of the MexAB-OprM efflux pump were evaluated. RESULTS: The percentages of susceptible isolates were as follows: ticarcillin, 62%; ticarcillin + clavulanic acid, 61%; piperacillin, 78%; piperacillin + tazobactam, 80% (MICs

[Campylobacter fetus bacteremia and cellulitis complicating a venous access port infection in an HIV infected patient]. / Bactériémie et dermohypodermite à Campylobacter fetus sur cathéter à chambre implantable au cours d'une infection par le VIH.

Campylobacter fetus subspecies fetus is an opportunist Gram-negative bacillus, which is known to be a cause of systemic infections, mainly in immunocompromised patients. We report a C. fetus bacteremia and cellulitis complicating a venous access port infection in a patient with acquired immunodeficiency syndrome (AIDS). This bacillus seems to have a predilection for the vascular endothelium and its isolation is difficult. Physicians should be aware of C. fetus infection in patients with vascular devices. Microbiologists should accurately isolate this organism from clinical specimens by modifying incubation techniques and performing molecular biology. The prognosis seems to be improved by a prolonged betalactam antibiotic regimen, especially amoxicilline plus clavulanic acid. In HIV infected patients, quinolones that were successful in our case, should be used with caution because of increasing resistance to antibiotics.

[Pasteurella dagmatis: an unusual cause of vertebral osteomyelitis]. / Pasteurella dagmatis: une cause inhabituelle de spondylodiscite.

Vertebral osteomyelitis is a quite rare but severe cause of back pain in adults. Various causative organisms have been reported. Pasteurella species have rarely been isolated. We report here a case of vertebral osteomyelitis and consecutive cauda equina syndrome due to Pasteurella dagmatis in a 60-year-old diabetic man.

[Community-acquired urinary tract infections in 15 to 65 years old female patients in France. Susceptibility of E. coli according to history: AFORCOPI-BIO network 2003]. / Infections urinaires communautaires de la femme de 15 à 65 ans: sensibilité aux antibiotiques de E. coli en fonction des antécédents: étude AFORCOPI-BIO 2003.

OBJECTIVES: A multicenter study was implemented in order to determine the distribution and antibiotic susceptibility patterns of strains isolated from 15 to 65 year old female patients with community-acquired urinary tract infections. PATIENTS AND METHODS: From October to December 2003, 11 French private laboratories consecutively collected 420 clinical strains with medical data. Minimal inhibitory concentrations of antibiotics on E. coli were determined using the agar dilution method in a coordinating center and interpretation followed the recommendations of the Comité de l'antibiogramme de la Société française de microbiologie. RESULTS: Escherichia coli was the most prevalent pathogen (80%) followed by Proteus mirabilis (4%), Klebsiella spp (2%), other Enterobacteriaceae (4%), Enterococcus spp (3%), Staphylococcus aureus (2%), Staphylococcus saprophyticus (2%), and Streptococcus agalactiae (2%). The susceptibility of E. coli strains was 61% for amoxicillin (AMX), 93% for nalidixic acid (NAL), 97% for norfloxacin (NOR) and ciprofloxacin (CIP), 77% for cotrimoxazole (SXT), 99% for fosfomycin, gentamicin and cefotaxime. The susceptibility of E. coli was lower in case of previous treatment with beta-lactam antibiotics for AMX (84 vs 95% p=0.02) and SXT (62 vs 81% p=0.02). In the same way, previous treatment with quinolones was associated with decreased susceptibility for NAL (84 vs 95% p=0.02) and SXT (62 vs 81% p=0.02). CONCLUSIONS: In 2003, fluoroquinolones, third generation cephalosporins, aminoglycosides, and fosfomycin kept a good activity on E. coli collected from community-acquired urinary tract infections in 15 to 65 years old female patients in France.

Occurrence of qnrA-positive clinical isolates in French teaching hospitals during 2002-2005.

Bacteria harbouring the novel qnrA plasmid-mediated mechanism of quinolone resistance have been described in different countries, but the frequency of their occurrence has not been investigated. In total, 1,468 clinical isolates of Enterobacteriaceae with quinolone resistance or extended-spectrum beta-lactamase (ESBL) phenotypes were collected from eight teaching hospitals in France during 2002-2005 and screened for qnrA. Overall, 28 isolates (22 Enterobacter cloacae, three Klebsiella pneumoniae, one Citrobacter freundii, one Klebsiella oxytoca and one Proteus mirabilis) were positive for qnrA, representing 1.9% of all isolates, 3.3% of ESBL-producing isolates (22% of the E. cloacae isolates) and 0% of non-ESBL-producing isolates. The prevalence of qnrA among consecutive ESBL-producing isolates in 2004 from the eight hospitals was 2.8% (18/639). Of the qnrA-positive isolates, 100% were intermediately-resistant or resistant to nalidixic acid, and 75% to ciprofloxacin. Twenty-one of the 22 qnrA-positive E. cloacae isolates were obtained from two hospitals in the Paris area, and molecular typing and plasmid content analysis showed clonal relationships for five, three and two isolates, respectively. The qnrA genetic environment was similar to that of the In36 integron. The remaining two isolates had qnrA variants (30 and 29 nucleotide differences, respectively, compared with the original sequence) and an unknown genetic environment. The ESBL gene associated with qnrA was bla(SHV-12) in most of the isolates, but bla(PER-1) and bla(SHV-2a) were found in two isolates. In France, it appears that qnrA-positive isolates are predominantly E. cloacae isolates producing SHV-12, and may be associated with the dissemination of an In36-like integron.

[Utility and limitations of laboratory diagnosis of amebiasis]. / Intérêt et limites du diagnostic au laboratoire de l'amibiase en 2004.

Entamoeba histolytica is an invasive and pathogenic protozoan parasite that causes amebiasis. It must be distinguished from Entamoeba dispar, a nonpathogenic commensal parasite of the human gut lumen that is morphologically identical to Entamoeba histolytica. Diagnosis of amoebic colitis currently requires combination of microscopic examination of stool specimens with another technique allowing positive identification of the two species. Stool culture followed by zymodem analysis is considered as gold standard but is not applicable in routine practice. Detection of specific Entamoeba histolytica antigens in stools is a fast, sensitive technique that should be considered as the method of choice. Stool PCR is a highly sensitive and specific technique but high cost make it unsuitable for use in endemic areas where economic conditions are difficult. The utility of serologic tests in distinguishing Entamoeba dispar from Entamoeba histolytica is controversial. However serology is still considered as the method of choice for diagnosis of extraintestinal amebiasis. Circulating Gal/GalNac lectin antigens can be detected in the serum of 96% of patients with untreated amoebic liver abscess. In the future this method should allow early diagnosis and treatment of extraintestinal amoebiasis in patients who have not yet developed detectable serum antibodies.

[Dermatological aspects in the risk of biological warfare]. / Aspects dermatologiques du risque biologique provoqué

The possible cutaneous manifestations of infectious biological warfare are multiple and vary depending on the agent used. An ulcerous and/or necrotic syndrome and/or regional lymphadenitis syndrome are possible with anthrax, tularaemia, bubonic plague and emission of trichotecene mycotoxins. A vesiculo-pustular syndrome with fever is provoked by smallpox, melioidosis and glanders. A purpural and/or haemorrhagic syndrome is seen during haemorrhagic fever viruses and septicaemic plague. These cutaneous manifestations are excellent markers that orient and alert when they occur in a context of a situation at risk, when several cases are observed in a usually non-exposed population and with extra-dermatological syndromes. They permit the early initiation of treatment.