ABSTRACT
Four strains isolated in the last 15 years were revealed to be identical in their 16S rRNA gene sequences to MCRO19, the sequence of which was deposited in GenBank in 1995. In a polyphasic analysis including phenotypic and genotypic features, the five strains (including MCRO19), which had been isolated in four European countries, turned out to represent a unique taxonomic entity. They are scotochromogenic slow growers and are genetically related to the group that included Mycobacterium simiae and 15 other species. The novel species Mycobacterium europaeum sp. nov. is proposed to accommodate these five strains. Strain FI-95228(T) (â=âDSM 45397(T) â=âCCUG 58464(T)) was chosen as the type strain. In addition, a thorough revision of the phenotypic and genotypic characters of the species related to M. simiae was conducted which leads us to suggest the denomination of the 'Mycobacterium simiae complex' for this group.
Subject(s)
Nontuberculous Mycobacteria/classification , Phylogeny , Adult , Aged, 80 and over , Bacterial Typing Techniques , Cell Wall/chemistry , DNA, Bacterial/genetics , Europe , Female , Genotype , Humans , Male , Molecular Sequence Data , Mycobacterium Infections/microbiology , Mycolic Acids/chemistry , Nontuberculous Mycobacteria/genetics , Nontuberculous Mycobacteria/isolation & purification , Phenotype , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
The polysaccharide capsule is a major virulence factor of Streptococcus pneumoniae as it confers resistance to phagocytosis. The encapsulated serotype 4 TIGR4 strain was shown to be efficiently phagocytosed by the mouse microglial cell line BV2, whereas the type 3 HB565 strain resisted phagocytosis. Comparing survival after uptake of TIGR4 or its unencapsulated derivative FP23 in gentamicin protection and phagolysosome maturation assays, it was shown that TIGR4 was protected from intracellular killing. Pneumococcal capsular genes were up-regulated in intracellular TIGR4 bacteria recovered from microglial cells. Actual presence of bacteria inside BV2 cells was confirmed by transmission electron microscopy (TEM) for both TIGR4 and FP23 strains, but typical phagosomes/phagolysosomes were detected only in cells infected with the unencapsulated strain. In a mouse model of meningitis based on intracranic inoculation of pneumococci, TIGR4 caused lethal meningitis with an LD(50) of 2 × 10² CFU, whereas the LD(50) for the unencapsulated FP23 was greater than 107 CFU. Phagocytosis of TIGR4 by microglia was also demonstrated by TEM and immunohistochemistry on brain samples from infected mice. The results indicate that encapsulation does not protect the TIGR4 strain from phagocytosis by microglia, while it affords resistance to intracellular killing.
Subject(s)
Bacterial Capsules/metabolism , Microbial Viability , Microglia/microbiology , Phagocytosis , Streptococcus pneumoniae/pathogenicity , Virulence Factors/metabolism , Animals , Bacterial Capsules/immunology , Brain/microbiology , Brain/pathology , Cells, Cultured , Disease Models, Animal , Gene Expression Profiling , Gene Expression Regulation, Bacterial , Genes, Bacterial , Immunohistochemistry , Lethal Dose 50 , Meningitis, Bacterial , Mice , Microglia/immunology , Microscopy, Electron, Transmission , Streptococcus pneumoniae/immunology , Survival Analysis , Virulence , Virulence Factors/immunologyABSTRACT
Clinical charts from 63 consecutive highly immunocompromised haematologic patients presenting with pulmonary nodular lesions on CT scan, classified as either probable or possible invasive fungal disease (IFD) according to the revised EORTC/MSG classification, were retrospectively studied. Histopathological analysis of lung tissues, available for 23 patients, demonstrated proven IFD in 17 cases (14 invasive aspergillosis and 3 invasive zygomycosis), diffuse alveolar damage in one and organising pneumonia (OP) in five cases. In the OP cases, three of which have been defined as probable IFD according to EORTC/MSG classification, extensive immunohistochemical, molecular and immunological analyses for fungi were negative. Our case descriptions extend the notion that OP may be encountered as a distinct histopathological entity in pulmonary nodular lesions in patients with leukaemia with probable/possible IFD.
Subject(s)
Cryptogenic Organizing Pneumonia/diagnosis , Cryptogenic Organizing Pneumonia/etiology , Leukemia, Myeloid, Acute/complications , Lung Diseases, Fungal/diagnosis , Lung Diseases, Fungal/etiology , Aged , Bronchoalveolar Lavage Fluid/microbiology , Diagnosis, Differential , Female , Humans , Immunocompromised Host , Invasive Pulmonary Aspergillosis/diagnosis , Invasive Pulmonary Aspergillosis/etiology , Leukemia, Myeloid, Acute/immunology , Male , Middle Aged , Opportunistic Infections/diagnosis , Opportunistic Infections/etiology , Retrospective Studies , Zygomycosis/diagnosis , Zygomycosis/etiologySubject(s)
Aspergillosis/etiology , Aspergillus/isolation & purification , Hyphae/ultrastructure , Leukemia, B-Cell/complications , Spleen/microbiology , Splenic Diseases/etiology , Acute Disease , Amphotericin B/therapeutic use , Antifungal Agents/therapeutic use , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Aspergillosis/drug therapy , Aspergillosis/pathology , Aspergillosis/surgery , Aspergillus/ultrastructure , Caspofungin , Combined Modality Therapy , Echinocandins/therapeutic use , Humans , Immunocompromised Host , Invasive Pulmonary Aspergillosis/complications , Invasive Pulmonary Aspergillosis/diagnosis , Leukemia, B-Cell/drug therapy , Lipopeptides , Male , Middle Aged , Neutropenia/chemically induced , Neutropenia/complications , Spleen/ultrastructure , Splenectomy , Splenic Diseases/drug therapy , Splenic Diseases/microbiology , Splenic Diseases/pathology , Splenic Diseases/surgery , Triazoles/therapeutic useABSTRACT
Although commercially available DNA probes for identification of mycobacteria have been investigated with large numbers of strains, nothing is known about the ability of these probes to identify less frequently encountered species. We analyzed, with INNO LiPA MYCOBACTERIA (Innogenetics) and with GenoType Mycobacterium (Hein), 317 strains, belonging to 136 species, 61 of which had never been assayed before. INNO LiPA misidentified 20 taxa, the majority of which cross-reacted with the probes specific for Mycobacterium fortuitum and the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum group. GenoType misidentified 28 taxa, most of which cross-reacted with M. intracellulare and M. fortuitum probes; furthermore, eight species were not recognized as members of the genus Mycobacterium. Among 54 strains investigated with AccuProbe (Gen-Probe), cross-reactions were detected for nine species, with the probes aiming at the M. avium complex being most involved in cross-reactions.