Showing papers on "Bartonella bacilliformis published in 2001"

Bartonella schoenbuchii sp. nov., isolated from the blood of wild roe deer.

Abstract: The genus Bartonella comprises two human-specific pathogens and a growing number of zoonotic or animal-specific species. Domesticated as well as wild mammals can serve as reservoir hosts for the zoonotic agents and transmission to humans may occur by blood sucking arthropods or by direct blood to blood contact. Humans may come into intimate contact with free-ranging mammals during hunting, especially during evisceration with bare hands, when accidental blood to blood contact frequently occurs. The objective of this work was to determine the presence and the polymorphism of Bartonella strains in wild roe deer (Capreolus capreolus) as the most widely spread game in Western Europe. We report the isolation of four Bartonella strains from the blood of five roe deer. These strains carry polar flagella similar to Bartonella bacilliformis and Bartonella clarridgeiae. Based on their phenotypic and genotypic characteristics, three of the four roe deer isolates were different and they were all distinct from previously described Bartonella species. They can be distinguished from each other and from other Bartonella species by their protein profile, ERIC-PCR pattern, 16S rRNA and citrate synthase (gitA) gene sequences, as well as by whole DNA-DNA hybridization. In spite of their considerable heterogeneity, all four strains fulfil the criteria for belonging to a single new species. The name Bartonella schoenbuchii is proposed for this new species. The type strain R1T of Bartonella schoenbuchii has been deposited in the National Collection of Type Cultures as NCTC 13165T and the Deutsche Sammlung von Mikroorganismen und Zellkulturen as DSM 13525T.

Establishing a Direct Role for the Bartonella bacilliformis Invasion-Associated Locus B (IalB) Protein in Human Erythrocyte Parasitism

Abstract: The invasion-associated locus A and B genes (ialAB )o fBartonella bacilliformis were previously shown to confer an erythrocyte-invasive phenotype upon Escherichia coli, indirectly implicating their role in virulence. We report the first direct demonstration of a role for ialB as a virulence factor in B. bacilliformis. The presence of a secretory signal sequence and amino acid sequence similarity to two known outer membrane proteins involved in virulence suggested that IalB was an outer membrane protein. To develop an antiserum for protein localization, the ialB gene was cloned in frame into an expression vector with a six-histidine tag and under control of the lacZ promoter. The IalB fusion protein was purified by nickel affinity chromatography and used to raise polyclonal antibodies. IalB was initially localized to the bacterial membrane fraction. To further localize IalB, B. bacilliformis inner and outer membranes were fractionated by sucrose density gradient centrifugation and identified by appearance, buoyant density (r), and cytochrome b content. Inner and outer membrane proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and IalB was positively identified by Western blot. Contrary to expectations, IalB was localized to the inner membrane of the pathogen. To directly demonstrate a role for IalB in erythrocyte parasitism, the B. bacilliformis ialB gene was disrupted by insertional mutagenesis. The resulting ialB mutant strain was complemented in trans with a replicative plasmid encoding the full-length ialB gene. PCR and high-stringency DNA hybridization confirmed mutagenesis and transcomplementation events. Abrogation and restoration of ialB expression was verified by SDS-PAGE and immunoblotting. In vitro virulence assays showed that mutagenesis of ialB decreased bacterial association and invasion of human erythrocytes by 47 to 53% relative to controls. Transcomplementation of ialB restored erythrocyte association and invasion rates to levels observed in the parental strain. These data provide direct evidence for IalB’s role in erythrocyte parasitism and represent the first demonstration of molecular Koch’s postulates for a Bartonella species.

Molecular phylogeny of the genus Bartonella: what is the current knowledge?

Abstract: Species of the genus Bartonella are involved in an increasing variety of human diseases. In addition to the 14 currently recognized species, several Bartonella strains have been recovered from a wide range of wild and domestic mammals in Europe and America. Such a high diversity of geographic distributions, animal reservoirs, arthropod vectors and pathogenic properties makes clarification of our knowledge about the phylogeny of Bartonella species necessary. Phylogenetic data have been inferred mainly from 16S rDNA, 16S–23S rRNA intergenic spacer, citrate synthase and 60 kDa heat-shock protein gene sequences, which are available in GenBank. Comparison of phylogenetic organizations obtained from various genes allowed six statistically significant evolutionary clusters to be identified. Bartonella bacilliformis and Bartonella clarridgeiae appear to be divergent species. Bartonella henselae, Bartonella koehlerae and Bartonella quintana cluster together, as well as Bartonella vinsonii subsp. vinsonii and B. vinsonii subsp. berkhoffii. The fifth group includes bacteria isolated from various rodents that belong to native species from the New World and in the sixth, Bartonella tribocorum, Bartonella elizabethae and Bartonella grahamii are grouped with several strains associated with Old World indigenous rodents. The position of the other species could not be consistently determined. As some cat- or rodent-associated Bartonella appeared to cluster together, it has been suggested that these bacteria and their reservoir hosts may co-evolve. Lack of host specificity, however, seems to be frequent and may reflect the influence of vector specificity. Host or vector specificity may also explain the current geographic distribution of Bartonella species.

Formation of stress fibres in human endothelial cells infected with Bartonella bacilliformis is associated with altered morphology, impaired migration and defects in cell morphogenesis.

Abstract: Bartonella bacilliformis, a Gram-negative, flagellated bacterium, infects human erythrocytes (haematic phase) and endothelial cells (tissue phase), resulting in a biphasic disease. In the tissue phase of disease (verruga peruana), infection leads to infection of endothelial cells and a pronounced proliferation of these cells, resulting in characteristic skin eruptions of papules and nodules. We have studied the properties of endothelial cells infected in vitro. Extensive cytoskeletal remodelling of endothelial cells occurred after infection in vitro with B. bacilliformis. The cells became spindle shaped and contained arrays of actin stress fibres orientated parallel to the long axis of the cell. Cell–cell contacts were disrupted, along with the distribution of the plasma membrane marker protein, PECAM-1, which participates in cell–cell junctions. The prominent stress fibres terminated in an increased number of focal contacts, which were studied using immunofluorescent staining for paxillin, a cytoplasmic protein that localizes in the focal adhesions. These morphological changes are consistent with activation of intracellular Rho by B. bacilliformis. Formation of stress fibres and the increased number of focal adhesions could be prevented by preincubation of the endothelial cells with C3 exoenzyme, which inactivates intracellular Rho by ADP ribosylation. Endothelial cell motility was greatly diminished in infected cells and the cells did not respond effectively to a stimulus that would evoke motility. In addition, infection of endothelial cells interfered with their ability to form networks of capillary tubes when suspended within three-dimensional collagen matrices. If the properties of infected endothelial cells in vivo are similar, the infected cells will probably not participate effectively in angiogenesis.

Deformin, a substance found in Bartonella bacilliformis culture supernatants, is a small, hydrophobic molecule with an affinity for albumin.

Abstract: Culture supernatants of Bartonella bacilliformis were previously shown to contain a factor, called deforming factor or deformin, which causes deformation and invagination of red cell membranes and formation of intracellular vacuoles. This factor is here shown to be a small water-soluble molecule, approximately 1400 Da as estimated by gel-filtration chromatography. Deforming factor binds tightly to albumin, especially albumin dimers and multimers, present in the growth medium. It can be released from albumin with 50% ethanol and has been partially purified by filtration and HPLC.