In vitro activity of temocillin against extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae strains isolated from urinary tract infections in France.

OBJECTIVES: Temocillin was introduced in 2015 in the French guidelines for the treatment of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae urinary tract infections. Little susceptibility data is available. We investigated the in vitro activity of temocillin against ESBL-producing Enterobacteriaceae isolated from samples of cytobacteriological examinations of urine. MATERIAL AND METHODS: Susceptibility testing was performed on 157 ESBL-producing E. coli and 95 ESBL-producing K. pneumoniae strains using the disk diffusion method. MICs of resistant strains were measured with the Etest method. RESULTS: Using current breakpoints, 71.3% of E. coli strains and 77.9% of K. pneumoniae strains were classified as susceptible. However, diameter and MIC breakpoints vary by country, and we reported discordance of clinical categorization between diameters and MIC determination for some strains. The measure of diameters was also sometimes difficult because of contaminating colonies within the inhibition zone. CONCLUSION: We highlighted difficulties related to the determination of temocillin susceptibility, such as culture of resistant colonies in the inhibition zone and discordance of clinical categorizations obtained with the disk diffusion method or the Etest method. Overall, 42% of tested Enterobacteriaceae had a diameter or MIC close to the current breakpoints; thus, it is necessary to determine the MIC for these strains before considering the clinical use of this molecule.

Trimethoprim susceptibility in E. coli community-acquired urinary tract infections in France.

OBJECTIVES: Trimethoprim has been recently included in the French guidelines for the treatment of urinary tract infections, but no epidemiological data supports its use. We aimed to determine the trimethoprim susceptibility of Escherichia coli isolates responsible for community-acquired urinary tract infections in women of childbearing age. MATERIALS AND METHODS: We conducted a national prospective survey. A total of 350 strains of E. coli isolated from urines in 35 laboratories were included. Antibiotic susceptibility testing was performed in each laboratory. RESULTS: We reported a susceptibility rate of 78%, and a similar clinical categorization between trimethoprim and cotrimoxazole for 97.4% of isolates. We pointed out an association between resistance to trimethoprim and other antibiotic classes. CONCLUSION: The results support trimethoprim as a second-line therapy based on antibiotic susceptibility testing results. We confirm that trimethoprim and cotrimoxazole susceptibility rates are very close.

Diagnosis of primary antibody and complement deficiencies in young adults after a first invasive bacterial infection.

OBJECTIVES: Screening for primary immunodeficiencies (PIDs) in adults is recommended after two severe bacterial infections. We aimed to evaluate if screening should be performed after the first invasive infection in young adults. METHODS: Eligible patients were retrospectively identified using hospital discharge and bacteriology databases in three centres during a 3-year period. Eighteen to 40-year-old patients were included if they had experienced an invasive infection with encapsulated bacteria commonly encountered in PIDs (Streptococcus pneumoniae (SP), Neisseria meningitidis (NM), Neisseria gonorrhoeae (NG), Haemophilus influenzae (HI), or group A Streptococcus (GAS)). They were excluded in case of general or local predisposing factors. Immunological explorations and PIDs diagnoses were retrieved from medical records. Serum complement and IgG/A/M testings were systematically proposed at the time of study to patients with previously incomplete PID screening. RESULTS: The study population comprised 38 patients. Thirty-six had experienced a first invasive episode and a PID was diagnosed in seven (19%): two cases of common variable immunodeficiency revealed by SP bacteraemia, one case of idiopathic primary hypogammaglobulinaemia, and two cases of complement (C6 and C7) deficiency revealed by NM meningitis, one case of IgG2/IgG4 subclasses deficiency revealed by GAS bacteraemia, and one case of specific polysaccharide antibody deficiency revealed by HI meningitis. Two patients had previously experienced an invasive infection before the study period: in both cases, a complement deficiency was diagnosed after a second NM meningitis and a second NG bacteraemia, respectively. CONCLUSION: PID screening should be considered after a first unexplained invasive encapsulated-bacterial infection in young adults.

In vitro susceptibility to mecillinam of Escherichia coli strains isolated from the urine of pregnant women.

OBJECTIVES: Pivmecillinam is a safe beta-lactam for use in pregnancy. It has been widely used for the treatment of lower urinary tract infections (UTIs) in the Nordic countries where its efficacy, minor impact on the microbiota, and low level of resistance among the Escherichia coli strains have been proven. However, susceptibility data related to E. coli involved in asymptomatic bacteriuria and lower UTIs in pregnant women is lacking. We aimed to support the 2015 recommendations issued by the French Infectious Diseases Society (SPILF) on gestational UTI, with a particular focus on pivmecillinam. MATERIAL AND METHODS: Antimicrobial susceptibility testing was performed by 12 hospitals with a maternity department on 235 E. coli strains isolated from the urine of pregnant women. Susceptibility to mecillinam was tested by disk diffusion method using the 2015 recommendations of the antibiogram committee of the French microbiology society (CA-SFM). RESULTS: Global susceptibility to mecillinam was 86.4%. Susceptibility to mecillinam was 96.5% for strains susceptible to amoxicillin-clavulanic acid and 38.7% for resistant strains. All six extended-spectrum beta-lactamase-producing E. coli strains were susceptible to mecillinam. CONCLUSION: Given the efficacy and safety of pivmecillinam during pregnancy, it may be used for the documented treatment of asymptomatic bacteriuria and acute cystitis in pregnant women. It also represents an alternative for the treatment of multidrug-resistant bacterial infections.

Evaluating the management of 493 patients presenting with bacteremia in 23 northern French hospitals.

OBJECTIVES: We aimed to update the epidemiology of bacteremia and evaluate their management and short-term outcome. METHODS: We conducted a prospective multicenter survey from October to November 2011. Consecutive patients with at least one positive blood culture (BC) were included in the study. We evaluated the type and adequacy of empirical and documented antibiotic therapy, time to active antibiotic therapy, compliance with guidelines, and 10-day outcome. RESULTS: A total of 23 public and private hospitals and 633 patients (493 true pathogens and 140 contaminants) were included in the study. Patients' wards were medicine (57%), surgery (19%), intensive care (14%), onco/hematology (3.7%), pediatrics (3.4%), infectious diseases (1.8%), and obstetrics (1.2%). Main pathogens were Escherichia coli (36%), Staphylococcus aureus (16%), coagulase-negative staphylococci, and Klebsiella sp. (8% each). A total of 43 (8.7%) multidrug-resistant strains were observed, including 26 extended-spectrum beta-lactamase strains and 15 methicillin-resistant S. aureus strains. An antibiotic active against the isolated pathogen was used in 74% of empirical and 96% of documented therapies. Median time between BC and administration of an active drug was 0.61 day. Empirical antibiotic therapies were protocol-compliant in 77% of cases. Few (4%) patients with contaminated BC received an antibiotic therapy (all inappropriate). Day-10 mortality was 12.1%, higher in patients presenting with severe sepsis or septic shock (22.5%) than in patients presenting with non-severe bacteremia (7.1%; P<0.0001). CONCLUSION: The management of bacteremia seems satisfactory in these volunteer hospitals but bacteremia remains a severe infection.

[Cluster of indigenous typhoid fever cases in a French city]. / Cas groupés de fièvre typhoïde autochtone dans une agglomération française.

BACKGROUND: A cluster of indigenous typhoid fever cases in the greater Lille area, in January 2009, triggered investigations to identify the contamination source and to optimize care for infected individuals. METHODS: A case was defined as a person, living in the greater Lille area of, having presented with symptoms of typhoid fever, from January to March 2009. RESULTS: Sixteen cases of typhoid fever were identified between January 23 and March 22, 2009. Patients, none of whom had travelled, had all participated in a common meal on January 10, 2009. A woman, who helped prepare the meal and who had previously stayed in an endemic zone, was detected as the asymptomatic carrier of Salmonella Typhi. CONCLUSION: In France, although typhoid fever remains essentially an imported disease, there is a risk of indigenous epidemic and its diagnosis can be suggested for individuals who have not travelled. The features of this cluster illustrate the importance of respecting basic rules of hygiene in catering.

[Antibiotics susceptibility of Streptococcus and Enterococcus: data of Onerba network]. / Sensibilité aux antibiotiques chez les streptocoques (hors pneumocoque) et les entérocoques: données Onerba.

This work was aimed to analyze trends in susceptibility to antibiotics among the main species of beta-hemolytic streptococci involved in community-acquired infections in human (Streptococcus pyogenes and Streptococcus agalactiae), or in animals (Streptococcus suis and Streptococcus uberis) and also among the main enterocci species, Enterococcus faecalis and Enterococcus faecium. Data were recorded since 1996 through the Onerba networks. S. pyogenes, as the other beta-hemolytic streptococci studied remained fully susceptible to beta-lactam antibiotics. However, susceptibility to macrolides is clearly decreasing in S. pyogenes. In 2002, only 62 to 65% of the strains according to the network considered, were susceptible to erythromycin. A similar trend was observed for S. agalactiae with only 75% of erythromycin susceptibility in 2002, and for both species isolated from animals S. suis and S. uberis, with respectively 35 and 76% of strains susceptible to erythromycin. In enterococci, susceptibility to beta-lactams remained stable between 2000 and 2004. Indeed, the susceptibility to aminopenicillins remained high in E. faecalis (about 98%), whereas the proportion of E. faecium isolates susceptible to these antibiotics were lower than 60%. From 1999 to 2004, various studies conducted in French hospitals showed that the vancomycin resistance among enterococci accounted for less than 2%. However, the recent emergence of glycopeptide resistant enterococci clusters in French hospitals is a matter of concern and emphasizes the need for an ongoing surveillance. Such trend in macrolide resistance among S. pyogenes or S. agalactiae should consequently lead to propose other alternatives in case of beta-lactam allergy, and for pharyngitis, to rethink the place of the culture for susceptibility testing.

Surveillance of methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE) in Northern France: a five-year multicentre incidence study.

In order to measure the incidence of methicillin-resistant Staphylococcus aureus (MRSA) and of Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE), and to evaluate the impact of the national guidelines for multidrug-resistant bacteria (MDRB) prevention in hospitals of Northern France, a multicentre study was conducted for three months every year starting in 1996, in volunteer hospital laboratories. All clinical specimens positive for MRSA and ESBLE were prospectively surveyed. During the five-year surveillance period, the overall proportion of MRSA was 38.4% in the 28,534 strains of S. aureus, and that of ESBLE was 11.4% in the 6121 strains of Klebsiella pneumoniae and 47.7% in the 2353 strains of Enterobacter aerogenes. The overall incidence rates of clinical specimens positive for MRSA, ESBL-K. pneumoniae and E. aerogenes were 0.84. 0.05 and 0.12/1000 hospital-days (HD), respectively. In the 23 hospitals that participated in the survey every year, the proportion and incidence of ESBLE decreased. Hence, despite recommendations as for isolation precautions, MRSA remains poorly controlled and requires more effective measures.

[Drug resistance in Mycobacterium tuberculosis: diagnostic methods]. / La résistance de Mycobacterium tuberculosis aux antituberculeux: méthodes diagnostiques.

The increasing frequency of multidrug-resistant strains of M. tuberculosis becomes dramatic in industrialized countries as well as in developing countries, particularly among patients infected with human immunodeficiency virus. It needs to formulate rapid strategies for diagnosing multidrug-resistant tuberculosis. For these new drug resistance, novel detection methods are developed in order to identify the resistant strains and to undertake efficacious antituberculosis therapies more rapidly. The phenotypic methods are based on the measurement of the microbial growth on nutritional supplement with antimicrobial agents; however, these proportional methods, such as the method in solid medium, the Bactec radiometric method or the MGIT method (mycobacterial growth indicator tube), are time consuming and give results in 5 to 21 days. In contrast, the genotypic tests, using knowledge of the genes involved in the resistance, reduce the time to detection of resistance from weeks to days. After amplification of the segment of the gene encoding the drug target by PCR, these methods are based on the identification of the different mutations conferring the antimicrobial resistance in M. tuberculosis. These methods are applied with success for detection of rifampicin resistance, conferring by mutations in a defined region of the rpoB gene for 99% of cases; on the contrary, results are less for other antituberculous drugs because of the insufficiency of knowledge of the molecular basis of drug resistance.

Molecular epidemiology of tuberculosis in the Nord Department of France during 1995.

In order to determine the current situation and to evaluate the human to human transmission of Mycobacterium tuberculosis in Northern France, the genetic polymorphism of strains was studied by using IS6110 fingerprint. One hundred and fifty-eight cases of bacteriologically confirmed tuberculosis were analyzed. One hundred and twenty-six patients (82%) were infected with genetically different isolates and 28 isolates (18%) were grouped into 14 clusters. No risk factors for recent Mycobacterium tuberculosis infections such as age, HIV status, immigrants, living in big cities were identified. This study shows that there was no major epidemic situation of tuberculosis in Northern France in 1995. Tuberculosis was characterized by a low proportion of HIV positive patients and a high proportion of elderly patients.

Identification of a new DNA region specific for members of Mycobacterium tuberculosis complex.

The successful use of DNA amplification for the detection of tuberculous mycobacteria crucially depends on the choice of the target sequence, which ideally should be present in all tuberculous mycobacteria and absent from all other bacteria. In the present study we developed a PCR procedure based on the intergenic region (IR) separating two genes encoding a recently identified mycobacterial two-component system named SenX3-RegX3. The senX3-regX3 IR is composed of a novel type of repetitive sequence, called mycobacterial interspersed repetitive units (MIRUs). In a survey of 116 Mycobacterium tuberculosis strains characterized by different IS6110 restriction fragment length polymorphisms, 2 Mycobacterium africanum strains, 3 Mycobacterium bovis strains (including 2 BCG strains), and 1 Mycobacterium microti strain, a specific PCR fragment was amplified in all cases. This collection included M. tuberculosis strains that lack IS6110 or mtp40, two target sequences that have previously been used for the detection of M. tuberculosis. No PCR fragment was amplified when DNA from other organisms was used, giving a sensitivity of 100% and a specificity of 100% in the confidence limit of this study. The numbers of MIRUs were found to vary among strains, resulting in six different groups of strains on the basis of the size of the amplified PCR fragment. However, the vast majority of the strains (approximately 90%) fell within the same group, containing two 77-bp MIRUs followed by one 53-bp MIRU.

[Tricentric study of the sensitivity of Pseudomonas aeruginosa serotyping to beta-lactams and aminoglycosides]. / Etude tricentrique de la sensibilité des sérotypes de Pseudomonas aeruginosa aux beta-lactamines et aux aminosides.

The aim of this study was to evaluate the susceptibility of 300 Pseudomonas aeruginosa strains to 6 beta-lactams and to 2 aminoglycosides and to compare this susceptibility according to the serotype, the sample nature, the unit activity and the geographic situation of the hospital. The susceptibility level was ascertained using the disk method. Serotyping of the strains was also performed. Susceptibility level was 67% for ticarcillin, 82% for piperacillin-tazobactam combination, 85% for ceftazidim, 81% for cefepim, 74% for aztreonam, 83% for imipenem, 73% for tobramycin and 86% for amikacin. Non-serotypable strains represented 20.6%. The most common serotypes were O6 (19%) and O11 (13%). Discrepancies between cefepime and ceftazidime were found in 15 strains after MIC determination, 6 of them were O12 strains. It appears that strain susceptibility was function of the serotype distribution. Cefepime exhibited outstanding activity against P. aeruginosa, similar to ceftazidim activity.

Antimicrobial activity among Pseudomonas and related strains of mineral water origin.

Siderophore production in 382 Pseudomonas and related strains of mineral water origin were screened and the antimicrobial activities of 158 of these tested against nine target organisms of health significance. Presence of siderophores could be detected in 54.4% and the majority of strains tested (91.2%) inhibited at least one of the nine target strains. Staphylococcus, Escherichia coli and Aeromonas hydrophila were particularly sensitive. Addition of iron eliminated the inhibitory activity in 96.7% of cases; the antagonistic effect should be largely determined by siderophore-mediated competition for iron. Most of the inhibitory strains produced siderophores, whereas the non-inhibitory strains did not. Few strains also produced bacteriocins showing activity against Pseudomonas aeruginosa and Aer. hydrophila. Strains isolated from mineral water have a broad antibacterial potential.

[Sensitivity of Pseudomonas aeruginosa to amikacin and to isepamicin in surgery and in intensive care]. / Sensibilité de Pseudomonas aeruginosa à l'amikacine et à l'isépamicine en chirurgie et en réanimation.

To evaluate the respective interest of amikacin and isepamicin in P. aeruginosa infection, the resistance level was ascertained using the disk method. Susceptibility was also tested for gentamicin, tobramycin and netilmicin. Isolates came from three surgical units and from two intensive care units. Serotyping was proceeded, and isolates coming from the same patient, with the same susceptibility pattern and the same serotype was included once only: 197 strains were thus obtained. Resistance level was 22.3% for amikacin, and 28.4% for isepamicin. Discrepancies were found in 14.7% of cases (major: 8.1%; minor: 6.6%). Discordant strains were more susceptible to amikacin than to isepamicin in 30/37 cases, and more susceptible to isepamicin in 7/37 cases. This difference was highly significant (paired Chi-2 test: p < 10(-4). The best susceptibility to amikacin was found in all serotypes and units.

[Preliminary study on tuberculin sensitivity in north Lebanon]. / Etude préliminaire de la sensibilité tuberculinique au Liban Nord.

Tuberculosis remains a severe public health problem, and is a major cause of infant disease in North Lebanon. We investigated tuberculin sensitivity in 1,405 school children (2 to 15 years old) not previously vaccinated with BCG in a deprived suburb of Tripoli (Lebanon). The Intra-Dermal Reaction (IDR) technique was used with 10 units of tuberculin (Pasteur-Mérieux, France). The prevalence was 4.3%. The score was higher in the lowest socio-economic group (5.7%) than in the middle socio-economic group (2.4%). No members of the upper socio-demographic group live in this area. There were no significant differences between the sexes. The diameter of induration was over 10 mm in 83.6% of cases (95.6% in the lower class and 46.6% in the middle class). The prevalence was higher among children over 10 years old. Chest X-rays were obtained for children with a positive reaction, and all were normal. Similarly, their sputum was tested for Acid Fast Bacilli (AFB) by a double staining technique (Ziehl-Nielsen and auramine), and all scored negative.

A blind study of the polymerase chain reaction for the detection of Mycobacterium tuberculosis DNA. Azay Mycobacteria Study Group.

SETTING: Nine French laboratories routinely involved in mycobacterial work. OBJECTIVE: To assess the detection of Mycobacterium tuberculosis in experimental samples by polymerase chain reaction (PCR) using the insertion sequence IS6110 as a target for deoxyribonucleic acid (DNA) amplification. DESIGN: Nine laboratories participated in a blind study of the detection of M. tuberculosis by PCR in 20 coded samples containing either a definite number of M. tuberculosis complex (positive samples) or environmental mycobacteria (four samples) or no mycobacteria (five samples). RESULTS: Five laboratories reported false-positive PCR results, with an average rate of 7%. All laboratories except one reported positive PCR results for samples containing 10(5) cfu/ml or more. M. tuberculosis DNA was detected in two thirds of samples containing 10(4) and 10(3) cfu/ml, and in one third of the samples containing 10(2) cfu/ml. CONCLUSION: The results of the study suggest that PCR using IS6110 as a target for DNA amplication is neither very sensitive nor really specific for the detection of M. tuberculosis.

[In vitro study of antibacterial activity of cefepime (Axepim) against gram negative bacteria: comparison with cephalosporins and other beta-lactams]. / Etude in vitro de l'activité antibactérienne du céfépime (axepim) sur les bacilles à gram négatif: comparaison avec les céphalosporines et les autres beta-lactamines.

The susceptibility to cefepime and to other beta-lactams of 1017 inducible cephalosporinase-producing enterobacteria, 897 Pseudomonas aeruginosa strains, and 295 Acinetobacter baumanii strains was studied over a two-year period (July 1, 1993 to June 30, 1995). The isolates were from patients in visceral surgery, intensive care, and clinical hematology wards. Cefepime was compared to other third-generation cephalosporins and to imipenem, aztreonam, and the piperacillin-tazobactam combination. Cefepime was more active than the other cephalosporins against Enterobacter cloacae, Serratia marcescens and Citrobacter freundii. Activity of cefepime on the study isolates was also greater than that of aztreonam and of the piperacillin-tazobactam combination. Cefepime exhibited outstanding activity against Pseudomonas aeruginosa strains except those with the O12 serotype.

Importance in diagnosis of gastritis of detection by PCR of the cagA gene in Helicobacter pylori strains isolated from children.

The cagA gene has been detected by PCR and DNA hybridization in 45 Helicobacter pylori strains isolated from children. For each child, clinical symptoms, endoscopic aspect of the gastric mucosa, and histological gastritis were evaluated. Gene-positive strains were associated with hemorrhagic gastritis in 66.6% of the children, while gene-negative strains were associated with hemorrhagic gastritis in 11.2% of the children (P = 0.0001). In addition, 88.8% of gene-positive strains were associated with severe histological gastritis (scores of 3 and 4), and gene-negative strains were collected from the gastric mucosa with the same type of infiltration of neutrophils and lymphocytes in the lamina propia in 55.5% of the children. These differences were statistically significant (P = 0.017). Gene-positive strains were also isolated more frequently from children with vomiting (P = 0.04), while the absence of clinical signs was not significantly different in cagA gene-positive or -negative patients. All of these observations confirmed the role of this cagA gene as a marker of gastric inflammation in children. The detection of this gene might be helpful to determine the degree of inflammation of the gastric mucosa in the absence of abdominal symptoms. We might better understand the natural history of H. pylori infection if we studied the evolution of gastritis in children with regard to the cagA status of isolated strains.

The retinoblastoma gene (RB-1) status in multiple myeloma: a report on 35 cases.

We looked for abnormalities of the retinoblastoma (RB-1) gene and of RB protein expression in 35 patients with multiple myeloma (MM). Mutations in exons 20 to 24 of the RB-1 gene (exons where mutations predominate in retinoblastoma and other solid tumors) were analyzed by single stranded conformation polymorphism (SSCP). RB-1 protein was studied in bone marrow plasma cells by immunocytochemistry (ABC peroxidase technique) with a specific monoclonal antibody. Southern blot analysis of RB-1 gene was also performed in 20 of the patients. Twenty two patients analyzed had advanced disease (stage III or, in one case, plasma cell leukemia) and cytogenetic analysis (performed in 31 cases) found monosomy 13 in 9 patients. No rearrangement of the RB-1 gene was found by Southern analysis. Absent or greatly reduced RB-1 protein level was found in plasma cells in 4 of the patients (11%), whereas normal levels were seen in the remaining cases. No point mutation in exons 20 to 24 and their flanking introns were found in any of the 35 patients. Three of the 4 patients with absent or reduced RB-1 protein expression had advanced MM (stage III: 2 cases; plasma cell leukemia: 1 case); all 4 patients were resistant to treatment (as compared to 7 of the 31 patients with normal RB-1 protein levels); only one of them was subsequently found to have monosomy 13 (as compared to 9 of the 28 other karyotyped patients). Our findings suggest that abnormalities of the RB-1 gene and its expression are rare in MM. Absent or reduced expression of RB-1 protein was not significantly correlated to monosomy 13 and was not associated with gross rearrangements of the RB-1 gene by Southern analysis or point mutations in exons 20 to 24 of the gene. Reduced expression of RB-1 protein may be associated with advanced disease and poor response to treatment, although larger numbers of patients will be required for more adequate conclusions.