Aims:  Aeromonas hydrophila is recognized as a human pathogen following wound exposure or ingestion of contaminated water and food. For rapid identification of this bacterium, a TaqMan‐based real‐time PCR assay has been developed. Methods and Results:  Primers and probes that target specific sequences of the 16S rRNA gene and cytolytic enterotoxin gene (aerA) were combined in a duplex assay. Presence and size of PCR products were confirmed with microchannel fluidics electrophoresis analysis. After validation, using type strain CIP7614T DNA, the PCR assay was tested on 12 positive and negative controls. Twenty‐one Aeromonas strains were isolated from environmental samples and were identified with biochemical tests as Aer. sobria, Aer. caviae and Aer. hydrophila. Only Aer. hydrophila strains tested positive by PCR assay. Conclusions:  The PCR developed here was successfully applied for the identification of Aer. hydrophila from reference, clinical and environmental samples and showed a high discrimination between Aer. hydrophila and other Aeromonas species. Significance and Impact of the Study:  This molecular method is convenient, rapid (2·5 h vs 24 h), specific to identify Aer. hydrophila and usable for diagnosis in medical and veterinary laboratories.

Staphylococcus aureus was isolated as the predominant or only isolate from cultures of stools of 60 patients over 2 years in a university hospital, leading to the collection of 114 isolates. Diarrhea was observed in 90% of the patients. Ninety-eight percent of the patients had received antibiotics in the month before the diarrhea. Ninety-two percent of the S. aureus isolates were methicillin resistant. S. aureus was encountered with antibiotic-associated diarrhea among 47 quite elderly patients affected or not affected by a gastrointestinal disease. Among the antimicrobial treatments, cessation of the previous therapy when possible or rapid application of oral vancomycin therapy was the most appropriate. Analysis of total DNA by pulsed-field gel electrophoresis revealed 27 different SmaI pulsotypes distributed in 15 clusters. The pulsotypes never differed for related isolates from a single patient, even if they originated from patients with bacteremia. S. aureus was not isolated as the predominant isolate in cultures of stools of 57 patients who received an antimicrobial treatment for more than 5 days without diarrhea. Occurence of production of both enterotoxin A and the bicomponent leucotoxin LukE-LukD by the S. aureus isolates was significantly different from that by random isolates. The results strongly suggest that when predominant in stool samples, S. aureus should be considered a possible etiologic agent for some cases of antibiotic-associated diarrhea.

Two Aeromonas strains, IBS S6874T and IBS S6652, were isolated from the faeces of two healthy monkeys (Macaca fascicularis) from Mauritius that were kept in quarantine in the Centre for Primatology, Strasbourg, France. Phylogenetic analysis based on 16S rRNA gene sequences showed that the two isolates formed an unknown genetic lineage within the genus Aeromonas.The two isolates had nearly identical sequences (0?1 % nucleotide substitution) that were related closely to those of recognized Aeromonas species (1?7-3?5 % nucleotide substitution). DNA-DNA hybridization showed that strains IBS S6874 T and IBS S6652 had high DNA-DNA similarity (89 %) to each other and a low level of DNA-DNA similarity to closely related taxa (18 % relatedness to Aeromonas trota and 16 % relatedness to Aeromonas schubertii ). Phenotypically, the two monkey isolates differed from most previously described mesophilic Aeromonas species by their lack of haemolysis on sheep-blood agar and inability to produce indole, gas from glucose or acid from mannitol. They differed from the most closely related species, A. schubertii, by their ability to produce acid from D-cellobiose and D-sucrose and by their pyrazinamidase activity. The name Aeromonas simiae sp. nov. is proposed for these isolates; strain IBS S6874 T (=CIP 107798is the type strain.The taxonomy of the genus Aeromonas has undergone continual change, due to reclassification or extended description of existing species and addition of newly described taxa (Huys et al., 1997a(Huys et al., , 2002Pidiyar et al., 2002;Esteve et al., 2003).Members of the genus Aeromonas are widespread in environmental habitats, such as soil (Brandi et al., 1996), fresh and brackish water, sewage and wastewater (Holmes et al., 1996). They are implicated as pathogens of coldblooded animals and various mammals, including humans, where they cause severe gastroenteritis, soft-tissue infections and bacteraemia (Janda & Abbott, 1998). Aeromonas isolates have been recovered from faeces in acute cases of gastroenteritis, as well as in asymptomatic cases (Hänninen & Siitonen, 1995). During a routine survey to determine the presence of possible pathogens in the intestinal tract of monkeys (Macaca fascicularis) from Mauritius that had been put in quarantine in the Centre for Primatology, Louis Pasteur University, Strasbourg, France, two phenotypically related strains, which were identified tentatively as Aeromonas sp., were isolated. These strains could not be identified as members of any previously described Aeromonas species. In the current investigation, phylogenetic analysis of 16S rRNA gene sequences, DNA-DNA hybridization and extensive phenotypic tests were undertaken to determine the taxonomic position of the two monkey isolates. Based on the reported data, we describe a novel species of the genus Aeromonas, for which the name Aeromonas simiae sp. nov. is proposed.Two strains (Institute of Bacteriology of Strasbourg strain IBS S6874T =CIP 107798 T =CCUG 47378 T and IBS S6652 =CIP 107797) were each isolated from a d...

We evaluated the incidence of Streptococcus pseudopneumoniae in clinical isolates by phenotypic methods and DNA-DNA hybridization. The pathogenic role of this organism was investigated with the mouse peritonitis/ sepsis model. Our results show a low incidence (1/120 pneumococcal isolates) and a potential pathogenic effect for S. pseudopneumoniae.Streptococcus pneumoniae, a major cause of morbidity and mortality worldwide, remains a significant health threat. Correct identification of pneumococci from clinical samples and differentiation from other oral streptococci are essential for appropriate diagnosis and treatment. A novel species of viridans group streptococci resembling S. pneumoniae was recently described as Streptococcus pseudopneumoniae (1); phenotypic identification is based on susceptibility to optochin (OPT) in ambient atmosphere and OPT resistance in a 5% CO 2 atmosphere, bile salt insolubility, and the absence of pneumococcal capsule. Genotypic identification is based on DNA-DNA hybridization, whereas sodA sequencing and 16S rRNA gene sequencing are not discriminating. While the new species tested positive for the pneumolysin gene (ply), its pathogenic role was not investigated. In the present study, we assessed the clinical significance of this new species by examining its incidence among pneumococci from clinical samples and investigating its virulence in a mouse peritonitis/sepsis model system.In total, 120 isolates presumptively identified as S. pneumoniae on the basis of Gram stain, catalase activity, and OPT susceptibility in ambient atmosphere were obtained from consecutive clinical samples of patients admitted to University Hospital, Strasbourg, France, during the period October 2004 to April 2005. Single pneumococcal isolates/patients/infections were analyzed. Eighteen isolates were collected from the lower respiratory tract, 57 were collected from the upper respiratory tract, 24 were collected from blood, 1 was collected from cerebrospinal fluid, 3 were collected from pleural fluid, and 12 were collected from other sterile body sites. Controls (S. pseudopneumoniae CCUG 49455 T and S. pneumoniae CCUG 28588 T ) were included in all assays. Additional clinical strains (S. pseudopneumoniae CCUG 48465, CCUG 50866, CCUG 50867, CCUG 50868, CCUG 50869, CCUG 50870, and CCUG 50871) described by Arbique et al. (1) were included in virulence assays.OPT susceptibility testing was performed by the disk diffusion method with a 6-mm disk (Bio-Rad) on sheep blood agar (Trypticase soy agar [bioMérieux] supplemented with 5% sheep blood); plates were incubated for 18 to 24 h at 35°C in ambient air and in a 5% CO 2 atmosphere. Solubility in bile salt (sodium deoxycholate [Merck]) was determined in tubes (4). Capsules were detected by observing a halo around pneumococci with India ink at a ϫ100 to ϫ400 magnification. Capsular agglutination tests were performed by the Pastorex test (BioRad) according to the instructions of the manufacturer.DNA-DNA hybridization was performed as described previously (8). Hybridi...

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A minilibrary of cationic N-heterocycles has been prepared and evaluated. The potential for the preparation was a result of the high versatility of palladium-mediated chemistry. The synthesis of the novel molecules was based on intramolecular quaternization of tertiary amine attached allylpalladium complexes. The steric and electronic factors of the reaction are discussed. The structures of the synthesized molecules made them candidates for precise biological and pharmaco-

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