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ABSTRACT: To assess the prevalence of efflux-driven fluoroquinolone (FQ) resistance in recent clinical isolates of Pseudomonas aeruginosa, a worrisome and often hospital-acquired pathogen, 115 unique strains were collected over a 5-month period, of which 27 and 33 had decreased susceptibility to ciprofloxacin (CIP) and levofloxacin (LVX), respectively. The MIC(50) (minimum inhibitory concentration for 50% of the organisms) was 16 μg/mL for both FQs. The efflux pump inhibitors (EPIs) phenylalanine-arginine-β-naphthylamide (PAβN) and 1-(1-naphthylmethyl)-piperazine (NMP) were then used to evaluate their efficacy in reducing CIP and LVX MICs. NMP did not significantly modify CIP MICs, whilst PAβN resulted in MIC(50) values of 2 μg/mL and 0.125 μg/mL for CIP and LVX, respectively. With the addition of PAβN, susceptibility to CIP and LVX was recovered in 6 (22.2%) and 31 (93.9%) strains, respectively. The best combination to reverse FQ resistance in this set of strains was LVX with PAβN. The results of this study show that the effect of an EPI is not only dependent on the species on which it is used but also on the molecule associated with it. Therefore, the design of an EPI equally efficient on all resistance-nodulation-cell division (RND) efflux pumps appears to be difficult and, from a practical point of view, if an EPI is developed for clinical use, the efficiency of its combination with a definite molecule should be assessed carefully against a wide range of clinical isolates to evaluate the real benefit of this combination.
International journal of antimicrobial agents 01/2012; 39(1):77-80. · 3.03 Impact Factor
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ABSTRACT: This study aimed at determining the contribution of intestinal bifidobacteria to the immune system activation using widely distributed galectins as markers of immune cell homoeostasis. In human flora-associated mice, bacteria were enumerated in the gut, blood, spleen, liver and lungs, while the expression of galectin-1 (Gal-1) and galectin-3 (Gal-3) was estimated by PCR in the intestine and real-time quantitative PCR in the other organs. Gal-1 and -3 were rarely expressed in the intestine. In blood, only Gal-1 was expressed while both galectins were expressed in all other organs. A high prevalence of colonic bifidobacteria was associated with a lower expression of both pulmonary galectins, whose levels negatively correlated with bifidobacterial counts. Caecal bifidobacterial counts also negatively correlated with pulmonary Gal-3 mRNA levels. The spleen was the only organ showing an upregulation of Gal-1 expression related to its bacterial contamination. However, this upregulation was only observed when bifidobacteria were not detected in the colon. A putative mechanism explaining the reduced expression of galectins when bifidobacteria highly colonize the mouse intestine could be that, by reducing the bacterial translocation, bifidobacteria also lead to a decreased blood concentration of substances produced by intestinal bacteria.
FEMS Immunology & Medical Microbiology 01/2009; 55(1):85-92. · 2.44 Impact Factor
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ABSTRACT: The aim of this work was to investigate the possible role of the intestinal anaerobic flora (especially bifidobacteria) in regulating bacterial translocation (BT) which can be defined as the passage of intestinal microbes through the mucosa to internal organs. Default in BT regulation concurs with pathogenesis of sepsis in various human conditions, such as acute pancreatitis, cirrhosis, necrotising enterocolitis or multiple organ failure. The intestinal flora was studied in human flora associated mice (HF mice) and BT was quantified in Peyer's patches (PP), blood, spleen, liver and lungs. HF mice displayed a heterogenic intestinal colonisation with bifidobacteria. High colonisation of both caecum and colon by bifidobacteria led to a poorer bacterial contamination of blood, liver and lungs. Moreover, ileal, caecal and colonic bifidobacterial counts negatively correlated with the bacterial dissemination (number of contaminated organs per mouse). In contrast, Bacteroides fragilis group counts positively correlated with bacteraemia, lungs contamination or bacterial dissemination. Additionally, clostridia localised in the colon affected bacterial uptake by PP and lungs contamination as indicated by positive correlations between bacterial populations in these respective locations. These results indicate that bifidobacteria, when established in high counts, reduced BT to liver, blood and lungs, whereas B. fragilis group favoured the bacterial passage. Clostridia established in the distal ileum also seemed to favour BT to lungs. The manipulation of the bacterial flora to optimise the regulatory effect on BT should therefore focus on the selective promotion of bifidobacteria and avoid an increase in potentially detrimental populations such as B. fragilis group and clostridia.
Anaerobe 03/2008; 14(1):43-8. · 2.41 Impact Factor
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ABSTRACT: One hundred and forty-four fluorescent pseudomonad strains isolated from various environments (soil, water, plant rhizosphere, hospital) and received as Pseudomonas putida (83 strains), P. putida biovar A (49 strains), P. putida biovar B (10 strains) and P. putida biovar C (2 strains), were analysed by the pyoverdine-isoelectrofocusing and pyoverdine-mediated iron uptake methods of siderotyping. Both methods demonstrated a great diversity among these strains, which could be subdivided into 35 siderovars. Some siderovars specifically included strains that have subsequently been transferred to well-defined Pseudomonas species, e.g. Pseudomonas monteilii or Pseudomonas mosselii, or which could be related by their siderotype to Pseudomonas jessenii or Pseudomonas mandelii. Other siderovars included strains sharing a high level of DNA-DNA relatedness (>70%), thus demonstrating that siderotyping could easily circumscribe strains at the species level. However, a group of seven strains, including the type strain, P. putida ATCC 12633T, were allocated into four siderovars, despite sharing DNA-DNA relatedness values of higher than 70 %. Interestingly, the strong genomic relationships between these seven strains were supported by the structural relationships among their pyoverdines, thus reflecting their phylogenetic affinities. These results strongly support the view that pyoverdine-based siderotyping could be used as a powerful tool in Pseudomonas taxonomy.
International journal of systematic and evolutionary microbiology 11/2007; 57(Pt 11):2543-56. · 2.27 Impact Factor
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ABSTRACT: Three multiplex polymerase chain reactions (PCRs) targeted on Bifidobacterium and related species were designed to identify human species. The selected primers yielded amplified products of various sizes, each specific for a species. Three to four pairs were gathered in one PCR reaction and their specificity under multiplex conditions was confirmed using DNA from 26 reference strains. Using this technique on unidentified faecal strains, B. bifidum, B. longum and B. breve species were commonly recovered in infants while B. adolescentis, B. catenulatum/B. pseudocatenulatum continuum and B. longum species were predominant in adults. Thus, a single PCR can provide the assignment of a strain to one these species, reducing the number of PCR reactions and hands-on time for the identification of human isolates of bifidobacteria. Moreover, this technique is also applicable for the in situ detection of bifidobacteria in DNA extracts from human stools.
FEMS Microbiology Letters 06/2003; 222(1):129-36. · 2.04 Impact Factor
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ABSTRACT: The vernacular name 'fluorescent Pseudomonas group 97-514' was coined for a group of 43 strains isolated from two French natural mineral waters. All these strains were gram-negative, rod-shaped and motile by means of a single polar flagellum. They produced fluorescent pigment (pyoverdin) on King B medium, catalase and cytochrome oxidase. They were capable of respiratory but not fermentative metabolism. They were not able to accumulate poly-beta-hydroxybutyrate and possessed an arginine dihydrolase system. DNA-DNA relatedness studies (S1 nuclease method) showed that the 43 strains of 'fluorescent Pseudomonas group 97-514' formed a genetically homogeneous group (DNA-DNA relatedness ranged from 70 to 100%). A total of 76 strains representing well-known or partially characterized species of the genus Pseudomonas sensu stricto had 7-56% DNA hybridization with strain CFML 97-514T. The highest DNA binding values were found with Pseudomonas veronii CIP 104663T (52%), Pseudomonas rhodesiae CIP 104664T (56%), Pseudomonas marginalis ATCC 10844T (56%), Pseudomonas gessardii CIP 105469T (53%) and Pseudomonas cedrella CIP 105541T (52%). Their unrelatedness was confirmed by deltaTm values greater than 7 degrees C. On the basis of the results of phenotypic and DNA-DNA hybridization studies, a novel Pseudomonas species, Pseudomonas grimontii sp. nov., is proposed for the 43 strains of 'fluorescent Pseudomonas group 97-514'. The type strain is strain CFML 97-514T (= CIP 106645T = ATCC BAA-140T). The G+C content of the DNA of the type strain was 58 mol%. A comparison of the complete 16S rRNA gene sequence of the type strain CFML 97-514T and the sequence of other strains of the genus Pseudomonas revealed that the novel species fell within the 'Pseudomonas fluorescens intrageneric cluster'. Members of P. grimontii grew at 4 degrees C but not at 41 degrees C. They were able to use D-xylose, alpha-L-rhamnose, alpha-aminobutyrate, meso-erythritol and itaconate as sole sources of carbon and energy and formed levan from sucrose. Strains do not possess lecithinase or Tween esterase activities. The clinical significance of P. grimontii is unknown.
International journal of systematic and evolutionary microbiology 10/2002; 52(Pt 5):1497-503. · 2.27 Impact Factor
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ABSTRACT: A total of 301 strains of fluorescent pseudomonads previously characterized by conventional phenotypic and/or genomic taxonomic methods were analyzed through siderotyping, i.e., by the isoelectrophoretic characterization of their main siderophores and pyoverdines and determination of the pyoverdine-mediated iron uptake specificity of the strains. As a general rule, strains within a well-circumscribed taxonomic group, namely the species Pseudomonas brassicacearum, Pseudomonas fuscovaginae, Pseudomonas jessenii, Pseudomonas mandelii, Pseudomonas monteilii, "Pseudomonas mosselii," "Pseudomonas palleronii," Pseudomonas rhodesiae, "Pseudomonas salomonii," Pseudomonas syringae, Pseudomonas thivervalensis, Pseudomonas tolaasii, and Pseudomonas veronii and the genomospecies FP1, FP2, and FP3 produced an identical pyoverdine which, in addition, was characteristic of the group, since it was structurally different from the pyoverdines produced by the other groups. In contrast, 28 strains belonging to the notoriously heterogeneous Pseudomonas fluorescens species were characterized by great heterogeneity at the pyoverdine level. The study of 23 partially characterized phenotypic clusters demonstrated that siderotyping is very useful in suggesting correlations between clusters and well-defined species and in detecting misclassified individual strains, as verified by DNA-DNA hybridization. The usefulness of siderotyping as a determinative tool was extended to the nonfluorescent species Pseudomonas corrugata, Pseudomonas frederiksbergensis, Pseudomonas graminis, and Pseudomonas plecoglossicida, which were seen to have an identical species-specific siderophore system and thus were easily differentiated from one another. Thus, the fast, accurate, and easy-to-perform siderotyping method compares favorably with the usual phenotypic and genomic methods presently necessary for accurate identification of pseudomonads at the species level.
Applied and Environmental Microbiology 07/2002; 68(6):2745-53. · 3.83 Impact Factor
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ABSTRACT: Twenty-two fluorescent pseudomonad strains of clinical origin received as Pseudomonas fluorescens (10 strains), Pseudomonasputida (10 strains) and Pseudomonas sp. (2 strains), and 33 type strains of the genus Pseudomonas were studied by numerical analysis based on 280 phenotypic characters. Twelve of the 22 clinical isolates clustered within a specific group, cluster IV. The other strains clustered within groups containing well-characterized fluorescent Pseudomonas species or did not cluster. Strains belonging to cluster IV were phenotypically different from all other clusters and subclusters of fluorescent pseudomonads. DNA-DNA hybridization showed that cluster IV corresponded to a genomic group sharing 72-100% DNA relatedness. DNA-DNA hybridization values with 67 strains representing 30 species of the genus Pseudomonas sensu stricto, including six recently described species (Pseudomonas veronii, Pseudomonas rhodesiae, Pseudomonas libanensis, 'Pseudomonas orientalis', 'Pseudomonas cedrella' and Pseudomonas monteilii), were below 49%, the value found for P. monteilii. The DNA G+C content of the type strain was 63 mol%. Comparison of the 16S rRNA gene sequence of a representative strain of cluster IV (CFML 90-83T) with sequences of other strains of the genus Pseudomonas revealed that strain CFML 90-83T was part of the P. fluorescens intrageneric cluster. On the basis of phenotypic, DNA-DNA hybridization and phylogenetic analyses, a novel species, Pseudomonas mosselii sp. nov., is proposed for the 12 strains of cluster IV. The type strain is P. mosselii CFML 90-83T (= ATCC BAA-99T = CIP 105259T). The P. mosselii strains are phenotypically homogeneous and can be differentiated from other fluorescent species by several phenotypic features, including pyoverdine typing.
International journal of systematic and evolutionary microbiology 04/2002; 52(Pt 2):363-76. · 2.27 Impact Factor
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ABSTRACT: The purpose of this study was to assess the epidemiology of local enterobacterial susceptibility to antibiotics. Between 1 January, 1998, and 31 December, 2001, we studied the sensitivity of 2,238 Enterobacteria to 26 different antibiotic agents in northern Lebanon, in the Microbiology department and Laboratory of the Islami Hospital, Tripoli, Lebanon. We used the diffusion disk method and complied with the guidelines of the French Microbiology Society antibiogram committee. Urinary samples were the most frequent source (67.5%), followed by blood cultures (12.7%). The dominant species in blood cultures was S. typhi (44.7%). We found 194 strains that produced extended-spectrum beta lactamases (ESBL), with the highest prevalence in Serratia spp. (44.3%), followed by Klebsiella pneumoniae (23.7%), Escherichia coli (20.7%) and Klebsiella oxytoca (11.3%). The global susceptibility of these strains to aminopenicillin was 15%; it reached 30% when combined with clavulanic acid. Susceptibility of the ESBL strains to these agents was 0%. The global susceptibility (and that of the ESBL strains, when greater than 0%) to other antibiotics was as follows: ticarcillin 38.5%, piperacillin 38.5%, piperacillin-tazobactam 88% (64%), imipenem 99.4%, (100%), cefalexin 41%, cefoxitin 65% (40.3%), cefuroxime 75%, amikacin 89%, chloramphenicol 30%, gentamicin 78% (42%), tetracycline 28% (16%), minocycline 30% (18.4%), colistin 67% (75%), nitrofuran 40% (45%), cotrimoxazol 40% (13%), nalidixic acid 53% (5.6%), pefloxacin 63% (23%), ciprofloxacin 71% (39%), and levofloxacin 72% (47%).
Sante (Montrouge, France) 13(2):107-12.
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ABSTRACT: The vernacular name ‘fluorescent Pseudomonas group 97-391’ was coined for a group of 11 strains isolated from two French natural mineral waters. All these strains were Gram-negative, rod-shaped and motile by means of a single polar flagellum. They produced fluorescent pigment (pyoverdin) on King B medium, catalase and cytochrome oxidase. They were not able to accumulate poly-β-hydroxybutyrate. They were capable of respiratory but not fermentative metabolism. DNA–DNA hybridization results and DNA base composition analysis revealed that strains of the ‘fluorescent Pseudomonas group 97-391’ were members of a new species, for which the name Pseudomonas brenneri sp. nov. (type strain CIP 106646T) is proposed. The levels of DNA–DNA relatedness within this group ranged from 70 to 100% with ΔTm below 1°C. The G+C content of the DNA of the type strain was 58 mol%. DNA relatedness with 72 strains representing well-known or partially characterized species of the genus Pseudomonas (sensu stricto) was below 48%. The complete 16S rRNA sequence of the type strain CIP 106646T was determined and compared with those of the type strains of Pseudomonas species. Finally, a phylogenetic tree was inferred from sequence analysis and demonstrated that the new species fell into the ‘Pseudomonas fluorescens intrageneric cluster’. The clinical significance of P. brenneri is unknown.
Research in Microbiology.
