To clarify the taxonomic status of two recently described Borrelia genomic groups, groups VS116 and M19, three group VS116 strains and eight group M19 strains isolated from Ixodes ricinus ticks in Switzerland, The Netherlands, and the United Kingdom were characterized. PCR-restriction fragment length polymorphism (RFLP) analysis of the 5s-23s intergenic spacer amplicon, rRNA gene restriction analysis, 16s rRNA gene sequence analysis, randomly amplified polymorphic DNA (RAPD) fingerprinting, sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotting with monoclonal antibodies were used for genetic and phenotypic analysis. The PCR-RFLP and RAPD patterns of three group VS116 strains and eight group M19 strains were identical but differed from those of Borrelia burgdorferi sensu stricto, Borrelia garinii, Borrelia afielii, and Borrelia japonica. DNAs from all group VS116 and M19 strains yielded three fragments (6.9, 3.2, and 1.4 kb) and four fragments (2.1, 1.2, 0.8, and 0.6 kb) after digestion with EcoRV and HindIII, respectively, hybridizing with an Escherichia coli 16s +23S cDNA probe. The SDS-PAGE protein profiles of group VS116 and M19 strains were heterogeneous. Phylogenetic analysis of the partial 16s rRNA gene sequences showed that group VS116 and M19 spirochetes were members of a Borrelia species distinct from previously characterized members of the genus Borrelia. Based on our present study and data from previous DNA-DNA hybridizations, a new Borrelia species, Borrelia valaisiana sp. nov., in the B. burgdorferi complex, is proposed. Strain VS116 is the type strain of this new species.

Corynebacterium ulcerans causes zoonotic infections, such as diphtheria and extrapharyngeal infections. We report here the first case of a diphtheria-like illness caused by C. ulcerans in France and transmitted likely by a dog to an immunocompromised woman. CASE REPORTA 47-year-old woman was admitted in the emergency room at the Bicêtre University Hospital (Le Kremlin-Bicêtre, France) for severe dyspnea in October 2003. She was immunocompromised due to treatment by prednisone at 6 mg/daily, tacrolimus at 12 mg/daily, and mycophenolate nofetil at 1 g/daily since she had undergone a kidney graft 1 year previously. Ten days prior to her admission to Bicêtre Hospital, the patient had been treated to no avail for sinusitis by oral amoxicillin-clavulanic acid (2 g/daily).Physical examination revealed severe stridor. She had fever at 38°C and mild tachycardia. The endoscopic examination identified a pseudomembranous exudate covering the nasopharynx and laryngeal vestibulum, ulceration, and subglottic constriction. A blood test found an increase of C-reactive protein (100 g/ml) and a white blood cell count of 3,500/l. This severe dyspnea indicated intubation, and the patient was subsequently hospitalized in the intensive care unit.Throat swab showed coryneform and gram-positive rods. After 24 h of incubation on sheep blood agar at 37°C in 5% CO 2 , shiny and whitish colonies grew that produced a slight hemolysis. Microscopic examination of these colonies revealed gram-positive, coryneform rods arranged in palisades. They were catalase positive, showed a positive reaction for urease, were negative for pyrazinamidase, and fermented glucose, ribose, maltose, and glycogen. These characteristics corresponded to those of Corynebacterium ulcerans. Biochemical identification was confirmed by partial sequencing of the 16S rRNA gene that showed 99% identity of the sequence of that strain with that of a C. ulcerans reference strain (11).Since C. ulcerans may acquire lysogenic ␤-corynephages coding for a diphtheria-like toxin (DT), PCR test (using primers DT1 and DT2 for detection of tox gene of C. diphtheriae (6) was performed that showed the presence of the DT-like gene in C. ulcerans. Disk diffusion susceptibility testing was performed on Mueller-Hinton blood agar (supplemented with 5% [vol/vol] sheep blood) after overnight incubation at 35°C and 5% CO 2 . In the absence of standardized breakpoints for Corynebacterium, antibiotic susceptibility was determined by using the NCCLS criteria for Streptococcus spp. other than Streptococcus pneumoniae (10).

We determined the complete sequence of the rrs gene from five strains of genomic species PotiB2. Both distance and parsimony methods were used to infer the evolutionary relationships of the rrs gene sequence of this genomic species in comparison with the rrs gene sequence of Borrelia valaisiana and the rrs gene sequences of BorreZia burgdogeri sensu lato species obtained from sequence databases. The phylogenetic analysis revealed that the genomic species PotiB2 strains clustered in a separate lineage, which was consistent with data from previous DNA-DNA hybridization experiments (D. Postic, M. V. ASSOUS, P. A. D. Grimont, and G. Baranton, Int. J. Syst. Bacteriol. 44:743-752, 1994). A PCR-restriction fragment length polymorphism analysis was used to identify genomic species PotiB2 and to differentiate it from B. burgdorferi sensu lato species. Moreover, signature nucleotide positions were identified for each B. burgdogeri sensu lato species. In accordance with DNA relatedness values, our findings suggest that genomic species PotiB2 can be more clearly defined and identified, and we propose that it should be referred to as a new species, BorreZia Zusitaniae. The type strain is PotiB2.Borrelia burgdorferi sensu lato, the causative agent of Lyme borreliosis, has been previously divided into five well-defined species, B. burgdorferi sensu stricto, Borrelia garinii, Borrelia afielii (1, 4), Borrelia japonica (12, 22), and Borrelia andersonii (17). The pathogenicity for humans of the first three species has been well-established, whereas the last two species seem not to be pathogenic for humans. These different species are not equally distributed all over the world, since the three main species, B. garinii, B. afzelii, and B. burgdorferi sensu stricto, occur in Europe, whereas B. burgdorferi sensu stricto is absent from Asia and is the main species encountered in the United States. In the United States, the more recently described species B. andersonii is also present as genomic species DN127, and each of the species is restricted to a limited geographical area. Additionally, in Japan, two species, Borrelia tanukii and Borrelia turdae, were described recently (6, 7). In Europe, two genomic species that are genetically and phenotypically divergent, genomic species VS116 and PotiB2, have been reported (20,21). The name Borrelia valaisiana is proposed in the accompanying paper for genomic species VS116 (29). Genomic species PotiB2 has been identified previously by DNA-DNA hybridization and has been characterized on the basis of the sequence of its qf-rrl ribosomal spacer (21). A specific MseI restriction pattern allowed workers to clearly identify this new genomic species, as well as all other delineated species or genomic species.The aim of this study was to further characterize genomic species PotiB2. First, characterization was completed at a genomic level by sequencing the rrs gene, which yielded an alternative means for specific identification. Second, the strains were studied at a phenotypic level with an analysis o...

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The goal of this study was to identify at the species level a large collection of Gram-negative dairy bacteria isolated from milks or semi-hard and soft, smear-ripened cheeses (cheese core or surface samples) from different regions of France. The isolates were then assessed for two risk factors, antibiotic resistance and volatile and non-volatile biogenic amine production in vitro. In total, 173 Gram-negative isolates were identified by rrs and/or rpoB gene sequencing. A large biodiversity was observed with nearly half of all Gram-negative isolates belonging to the Enterobacteriaceae family. Overall, 26 different genera represented by 68 species including potential new species were identified among the studied Gram-negative isolates for both surface and milk or cheese core samples. The most frequently isolated genera corresponded to Pseudomonas, Proteus, Psychrobacter, Halomonas and Serratia and represented almost 54% of the dairy collection. After Pseudomonas, Chryseobacterium, Enterobacter and Stenotrophomonas were the most frequently isolated genera found in cheese core and milk samples while Proteus, Psychrobacter, Halomonas and Serratia were the most frequently isolated genera among surface samples. Antibiotic resistance profiles indicated that resistances to the aminosid, imipemen and quinolon were relatively low while more than half of all tested isolates were resistant to antibiotics belonging to the monobactam, cephem, fosfomycin, colistin, phenicol, sulfamid and some from the penam families. Thirty-six% of isolates were negative for in vitro biogenic amine production. Among biogenic amine-producers, cadaverine was the most frequently produced followed by isoamylamine, histamine and putrescine. Only low levels (<75 mg/l) of tyramine were detected in vitro.

Aims: To isolate, identify, and characterize heterotrophic bacteria in acid-mine drainage that mediate oxidation of As(III). Methods and Results: Samples of acid-mine drainage were collected over a period of 14 months. Heterotrophic and non-obligatory acidophilic bacteria in the samples were cultured on a solid medium (pH 7AE0-7AE2), and three strains were isolated. The three different strains belong to the genus Thiomonas, and have more than 99% homology with the group Ynys1. Culturing in mineral media demonstrated that the isolated strains used thiosulphate as an energy source, and oxidized iron in the presence of thiosulphate. However, none of the strains were able to oxidize arsenic in the presence of thiosulphate, nor could they use iron or arsenic alone as an energy source. In vitro experiments demonstrated that two of the Thiomonas strains were able to oxidize more than 90% of the As(III) present in the acid-mine drainage, whereas no abiotic oxidation of arsenic occurred. Conclusions: Two strains of newly identified Thiomonas sp. found in acid-mine drainage are capable of oxidizing arsenic. Significance and Impact of Study: These results represent the first reported oxidation of arsenic by Thiomonas sp. Biologically mediated oxidation and subsequent immobilization of arsenic is of great interest for the remediation of contaminated mine sites.

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...

Acinetobacter ursingii has not been reported in infectious processes apart from its recent description as a new species. A bacteremia caused by A. ursingii in a patient with a pulmonary adenocarcinoma confirms that this microorganism is an opportunistic human pathogen. The isolate was susceptible to imipenem, aminoglycosides, rifampin, and fluoroquinolones. CASE REPORTA 63-year-old man presenting with a pulmonary adenocarcinoma diagnosed in July 2001 was admitted to the University Hospital Hôtel Dieu, Paris, France. A fifth course of intravenous chemotherapy consisting of cisplatin (50 mg/m 2 of body area) and vinorelbine (30 mg/m 2 ) was initiated through a catheter chamber, which had been implanted 12 weeks earlier. The patient had received corticosteroids for 2 months because of thoracic pain due to tumoral compression. On the day following admission (day 1), his condition deteriorated with fever (39.5°C) and chills associated with an increase of white blood cell count (15 ϫ 10 3 cells per l with 90% neutrophils), C-reactive protein (9.5 mg/dl), and fibrinogen (0.51 mg/dl). Thus, he received cefotaxime (3 g per day) and gentamicin (180 mg per day) for 2 days. Urine analysis was normal. The chest X ray and the abdominal and cardiac echography were not modified. A strain of an Acinetobacter sp. was isolated from five blood samples, including two obtained from the catheter chamber. The catheter chamber was removed, but its culture was sterile. On day 3, the antimicrobial therapy was changed to imipenem (2 g per day) and amikacin (900 mg per day) for 2 weeks according to the susceptibility of the strain. On day 5, rifampin (1.2 g per day) was added as the patient remained febrile. On day 7, the patient was apyretic, with normalization of white blood cell count and C-reactive protein.Blood samples were inoculated in aerobic and anaerobic blood culture vials (BACTEC PLUS; BD Diagnostic Systems, Sparks, Md.). Aerobic vials were positive and were subcultured on nutrient agar at 37°C. After 24 h of incubation, colonies were 1 to 1.5 mm in diameter, circular, convex, smooth, and slightly opaque with entire margins. Staining of the bacteria showed gram-negative coccobacilli. Growth in brain heart infusion (BHI) broth was observed at 37°C but not at 41 and 44°C. The microorganism (isolate 954) was nonmotile, strictly aerobic, and oxidase negative. It grew on MacConkey agar (colorless colonies), was nonhemolytic on sheep blood agar, did not oxidize D-glucose, did not reduce nitrate, and was urease and gelatinase negative. In an attempt to identify this isolate, the strips API 20 NE and API ID 32 GN (bioMérieux, Marcy l'Etoile, France) were used as recommended by the manufacturer. The repeated bacterial identifications obtained with API 20 NE and API ID 32 GN strips were Acinetobacter junii or Acinetobacter johnsonii (code no. 0000071; percentage of identification [p] ϭ 63.5%; index of typicity [T] ϭ 0.77) and A. johnsonii (code no. 00270063062; p ϭ 90.5%; T ϭ 0.87), respectively. These results prompted us to determine the...