Claire Hennequin

Lactobacillus rhamnosus Meningitis following Recurrent Episodes of Bacteremia in a Child Undergoing Allogeneic Hematopoietic Stem Cell Transplantation

ABSTRACT We report a case of meningitis due to Lactobacillus rhamnosus in a child undergoing allogeneic hematopoietic stem cell transplantation for acute leukemia. Four episodes of bacteremia involving strains with pulsotypes identical to that of the cerebrospinal fluid isolate preceded meningitis. After several courses of clindamycin, no relapse occurred during the patient follow-up.

Immunization Using GroEL Decreases Clostridium difficile Intestinal Colonization

Clostridium difficile is a pathogen which is responsible for diarrhea and colitis, particularly after treatment with antibiotics. Clinical signs are mainly due to two toxins, TcdA and TcdB. However, the first step of pathogenesis is the colonization process. We evaluated C. difficile surface proteins as vaccine antigens in the hamster model to prevent intestinal colonization. This vaccination induced a partial protection of hamsters against death after a C. difficile challenge. A proteomic analysis of animal sera allowed us to identify proteins which could be responsible for the protection observed. Among these proteins, we identified the GroEL heat shock protein. To confirm the role of the specific GroEL antibodies in the delayed C. difficile colonization of hamsters, we performed an immunization assay in a mouse model. After intranasal immunization with the recombinant protein GroEL, we observed a lower C. difficile intestinal colonization in the immunized group as compared to the control group.

Assessment on Experimental Bacterial Biofilms and in Clinical Practice of the Efficacy of Sampling Solutions for Microbiological Testing of Endoscopes

ABSTRACT Opinions differ on the value of microbiological testing of endoscopes, which varies according to the technique used. We compared the efficacy on bacterial biofilms of sampling solutions used for the surveillance of the contamination of endoscope channels. To compare efficacy, we used an experimental model of a 48-h Pseudomonas biofilm grown on endoscope internal tubing. Sampling of this experimental biofilm was performed with a Tween 80-lecithin-based solution, saline, and sterile water. We also performed a randomized prospective study during routine clinical practice in our hospital sampling randomly with two different solutions the endoscopes after reprocessing. Biofilm recovery expressed as a logarithmic ratio of bacteria recovered on bacteria initially present in biofilm was significantly more effective with the Tween 80-lecithin-based solution than with saline solution (P = 0.002) and sterile water (P = 0.002). There was no significant difference between saline and sterile water. In the randomized clinical study, the rates of endoscopes that were contaminated with the Tween 80-lecithin-based sampling solution and the saline were 8/25 and 1/25, respectively (P = 0.02), and the mean numbers of bacteria recovered were 281 and 19 CFU/100 ml (P = 0.001), respectively. In conclusion, the efficiency and therefore the value of the monitoring of endoscope reprocessing by microbiological cultures is dependent on the sampling solutions used. A sampling solution with a tensioactive action is more efficient than saline in detecting biofilm contamination of endoscopes.

Streptobacillus moniliformis as the Causative Agent in Spondylodiscitis and Psoas Abscess after Rooster Scratches

ABSTRACT We report a case of Streptobacillus moniliformis spondylodiscitis accompanied by a psoas abscess in an 80-year-old man scratched by a rooster. S. moniliformis was identified from abscess fluid by use of 16S rRNA gene sequencing. After 18 weeks of antimicrobial therapy, the clinical condition of the patient improved.

Rapid detection of AAC(6’)-Ib-cr production using a MALDI-TOF MS strategy

Plasmid-mediated quinolone resistance mechanisms have become increasingly prevalent among Enterobacteriaceae strains since the 1990s. Among these mechanisms, AAC(6’)-Ib-cr is the most difficult to detect. Different detection methods have been developed, but they require expensive procedures such as Sanger sequencing, pyrosequencing, polymerase chain reaction (PCR) restriction, or the time-consuming phenotypic method of Wachino. In this study, we describe a simple matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) method which can be easily implemented in clinical laboratories that use the MALDI-TOF technique for bacterial identification. We tested 113 strains of Enterobacteriaceae, of which 64 harbored the aac(6’)-Ib-cr gene. We compared two MALDI-TOF strategies, which differed by their norfloxacin concentration (0.03 vs. 0.5 g/L), and the method of Wachino with the PCR and sequencing strategy used as the reference. The MALDI-TOF strategy, performed with 0.03 g/L norfloxacin, and the method of Wachino yielded the same high performances (Se = 98 %, Sp = 100 %), but the turnaround time of the MALDI-TOF strategy was faster (<5 h), simpler, and inexpensive (<1 Euro). Our study shows that the MALDI-TOF strategy has the potential to become a major method for the detection of many different enzymatic resistance mechanisms.

Chorioamniotite à Streptococcus anginosus et Streptococcus gallolyticus subsp. pasteurianus

Une femme de 34 ans a été hospitalisée pour une enace d’accouchement prématuré (MAP) sévère à 24 semaines ’aménorrhée (SA) sur une grossesse gémellaire. La patiente e présentait pas de contraction, mais la poche des eaux était alpable dans le vagin. En prévention du risque de chorioamiotite, une antibiothérapie probabiliste a été mise en place : 40 mg de gentamicine (Gentalline®) une fois par jour en intrausculaire et 1 g de céfotaxime (Claforan®) trois fois par jour en ntraveineux. Une corticothérapie (bétaméthasone, Célestène®) e maturation pulmonaire a été débutée et une tocolyse a été nitiée (nicardipine, Loxen® au pousse-seringue). De faibles ontractions ont cependant débuté. Malgré un second traiteent tocolytique (atosiban, Tractocile®), la poche des eaux s’est etrouvée extériorisée et bombante au-delà de la vulve associée une métrorragie faible. Une dilatation complète a été obteue en quatre heures. La rupture de la poche des eaux a eu lieu ors du travail et l’accouchement par voie basse s’est déroulé apidement en une heure. Le liquide amniotique ne présentait ucune anomalie notable, mais les deux nouveau-nés étaient ort-nés. La numération formule a montré une hyperleucocyose modérée ; le bilan inflammatoire a révélé un fibrinogène à ,9 g/l et une CRP à 6,1 mg/l. L’examen anatomopathologique es deux placentas a révélé une chorioamniotite sur le placenta a d à d

Diversity of DHA-1-encoding plasmids in Klebsiella pneumoniae isolates from 16 French hospitals

Objectives To provide new insights into the spread of plasmidic cephalosporinase DHA-1, 16 strains of Klebsiella pneumoniae and a strain of Klebsiella variicola producing DHA-1 were isolated between January 2012 and December 2013 in six regions of France and two French overseas departments and territories. Methods Disc diffusion assays, isoelectric focusing and PCRs were used to characterize the plasmidic DHA-1 β-lactamase. Plasmid analysis was performed by the method of Kado and Liu and WGS. Virulence of the strains was studied by biofilm formation and the survival of Drosophila. Results The strains were of low virulence and had one to three plasmids including one of various sizes (∼40 to 319 kb) mediating DHA-1. Nine strains belonged to ST11 and possessed a pKPS30-type DHA-1 plasmid of the IncR (incompatibility) group. A strain of ST307 possessed pENVA, a DHA-1 plasmid of the IncH-type group. The seven remaining plasmids were unknown. Three belonged to the IncL/M group. They were closely related and their sequences were determined. One of the four remaining strains was chosen for further investigation. This strain of ST16 had two plasmids, a pUUH239.2-related plasmid and a new DHA-1 plasmid of ∼319 kb of IncHI2 type. Conclusions These findings demonstrate the major role of the pKPS30-type plasmid in the spread of DHA-1 cephalosporinase in France and provide evidence of two new emerging plasmids carrying this enzyme.