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1.
Diagn Microbiol Infect Dis ; 99(1): 115201, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33065460

ABSTRACT

We evaluated the performance of three chromogenic media (BBL CHROMagar™ Staph aureus, ChromID™ S. aureus SAID, ChromID™ S. aureus Elite SAIDE) for the isolation of Staphylococcus aureus in respiratory samples in patients with cystic fibrosis in comparison with CNA media. We reported a similar ability of the four media to support the growth of S. aureus and that sensitivity increased when incubation lasted more than 24 h. SAIDE had the higher sensitivity compared to the other media and kept a high specificity even after 72 h.


Subject(s)
Chromogenic Compounds/pharmacology , Culture Media/pharmacology , Cystic Fibrosis/microbiology , Staphylococcus aureus/growth & development , Staphylococcus aureus/isolation & purification , Bronchoalveolar Lavage Fluid/microbiology , Culture Media/chemistry , Humans , Lung/microbiology , Sputum/microbiology , Staphylococcal Infections/microbiology , Trachea/microbiology
2.
Front Microbiol ; 7: 1991, 2016.
Article in English | MEDLINE | ID: mdl-28018320

ABSTRACT

The multi-resistant Staphylococcus capitis clone NRCS-A has recently been described as a major pathogen causing nosocomial, late-onset sepsis (LOS) in preterm neonates worldwide. NRCS-A representatives exhibit an atypical antibiotic resistance profile. Here, the complete closed genome (chromosomal and plasmid sequences) of NRCS-A prototype strain CR01 and the draft genomes of three other clinical NRCS-A strains from Australia, Belgium and the United Kingdom are annotated and compared to available non-NRCS-A S. capitis genomes. Our goal was to delineate the uniqueness of the NRCS-A clone with respect to antibiotic resistance, virulence factors and mobile genetic elements. We identified 6 antimicrobial resistance genes, all carried by mobile genetic elements. Previously described virulence genes present in the NRCS-A genomes are shared with the six non-NRCS-A S. capitis genomes. Overall, 63 genes are specific to the NRCS-A lineage, including 28 genes located in the methicillin-resistance cassette SCCmec. Among the 35 remaining genes, 25 are of unknown function, and 9 correspond to an additional type I restriction modification system (n = 3), a cytosine methylation operon (n = 2), and a cluster of genes related to the biosynthesis of teichoic acids (n = 4). Interestingly, a tenth gene corresponds to a resistance determinant for nisin (nsr gene), a bacteriocin secreted by potential NRCS-A strain niche competitors in the gut microbiota. The genomic characteristics presented here emphasize the contribution of mobile genetic elements to the emergence of multidrug resistance in the S. capitis NRCS-A clone. No NRCS-A-specific known virulence determinant was detected, which does not support a role for virulence as a driving force of NRCS-A emergence in NICUs worldwide. However, the presence of a nisin resistance determinant on the NRCS-A chromosome, but not in other S. capitis strains and most coagulase-negative representatives, might confer a competitive advantage to NRCS-A strains during the early steps of gut colonization in neonates. This suggests that the striking adaptation of NRCS-A to the NICU environment might be related to its specific antimicrobial resistance and also to a possible enhanced ability to challenge competing bacteria in its ecological niche.

3.
J Leukoc Biol ; 92(6): 1241-50, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23033174

ABSTRACT

The migration of DCs is a critical function, enabling information to be carried to where the immunological response occurs. Parasites are known to weaken host immunity by interfering with the functions of DCs and thus, may be a source of molecules with immunomodulatory properties. Here, we demonstrate that the soluble protein, GRA5, specific to Toxoplasma gondii, is able to increase the migration of human CD34-DCs toward CCL19. A synthetic Pep29 derived from the GRA5 hydrophilic NT region (Pep29) was found to be internalized by macropinocytosis and to trigger in vitro migration of CD34-DCs via CCR7 expression without activating DCs. Pep29 also induced a decrease in the number of LCs from human skin epidermis. As local depletion of DCs and migration of immature DCs lead to a disruption of the specific innate response, our results highlight the potential of using pathogen-derived synthetic peptides as novel cell modulators with a therapeutic potential to reduce symptoms in inflammatory disorders.


Subject(s)
Antigens, Protozoan/pharmacology , Cell Movement/drug effects , Dendritic Cells/drug effects , Peptides/pharmacology , Toxoplasma/immunology , Amino Acid Sequence , Antigens, CD34/metabolism , Antigens, Protozoan/chemistry , Antigens, Protozoan/immunology , Antigens, Protozoan/metabolism , Cell Movement/immunology , Cells, Cultured , Chemokine CCL19/immunology , Chemokine CCL19/metabolism , Dendritic Cells/immunology , Dendritic Cells/metabolism , Humans , Immunophenotyping , Langerhans Cells/immunology , Langerhans Cells/metabolism , MAP Kinase Signaling System , Molecular Sequence Data , Peptides/chemistry , Peptides/immunology , Peptides/metabolism , Pinocytosis/immunology , Protein Interaction Domains and Motifs/immunology , Protein Sorting Signals , Protozoan Proteins/chemistry , Protozoan Proteins/immunology , Receptors, CCR7/metabolism , Skin/immunology , Toxoplasma/chemistry
4.
J Microbiol Methods ; 86(2): 145-9, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21545818

ABSTRACT

Flow cytometry is a powerful tool for analyzing the adhesion to and invasion of Staphylococcus aureus (S. aureus) to eukaryotic cells. Established techniques have used bacteria that have been genetically modified to express fluorescent proteins or directly labeled with fluorochromes prior to infection. Such approaches are appropriate in most cases; however, the use of genetically or chemically altered bacteria could introduce a bias when measuring fine differences in adhesion and invasiveness. Here, we describe a combined flow cytometry-based invasion and adhesion assay that does not require the processing of bacteria prior to internalization. This method was performed on osteoblastic MG-63 cells infected with S. aureus reference strain 8325-4 and its invasion-deficient isogenic mutant, which carries deletions in the genes encoding fibronectin-binding proteins A and B. The data from this assay were compared to those obtained using the standard gentamicin protection assay. The results obtained by the two methods were consistent. Moreover, quantification of internalized bacteria was more reproducible using the flow cytometry-based assay than the gentamicin protection assay, which allowed for the simultaneous quantification of host cell adhesion and invasion.


Subject(s)
Bacterial Adhesion , Flow Cytometry/methods , Osteoblasts/microbiology , Staphylococcus aureus/pathogenicity , Adhesins, Bacterial/genetics , Cell Line , Gene Deletion , Humans , Staphylococcus aureus/genetics , Virulence Factors/genetics
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