The Biology of Hemotrophic Bacteria

Kreier, Julius P.; Ristic, M

doi:10.1146/annurev.mi.35.100181.001545

Cited by 39 publications

(29 citation statements)

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“…Only one isolate of Bartonella elizabethae has been obtained from the heart valve of a patient with endocarditis (5). Finally, Bartonella bacilliformis is responsible for bartonellosis (15).…”

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Use of rpoB Gene Analysis for Detection and Identification of Bartonella Species

et al. 2001

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Identification of Bartonella species is of increasing importance as the number of infections in which these bacteria are involved increases. To date, these gram-negative bacilli have been identified by various serological, biochemical, and genotypic methods. However, the development of alternative tools is required, principally to circumvent a major risk of contamination during sample manipulation. The aim of our study was to investigate the possible identification of various Bartonella species by comparison of RNA polymerase beta-subunit gene (rpoB) sequences. This approach has previously been shown to be useful for the identification of members of the family Enterobacteriaceae (C. M. Mollet, M. Drancourt, and D. Raoult, Mol. Microbiol. 26:1005-1011, 1997). Following PCR amplification with specific oligonucleotides, a 825-bp region of the rpoB gene was sequenced from 13 distinct Bartonella strains. Analysis of these sequences allowed selection of three restriction enzymes (ApoI, AluI, and AflIII) useful for discerning the different strains by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis. To confirm the potential value of such an approach for identification of Bartonella, the rpoB PCR was then applied to 94 clinical samples, and the results obtained were identical to those obtained by our reference PCR method. Twenty-four isolates were also adequately identified by PCR-RFLP analysis. In all cases, our results were in accordance with those of the reference method. Moreover, conserved regions of DNA were chosen as suitable primer targets for PCR amplification of a 439-bp fragment which can be easily sequenced.

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Use of rpoB Gene Analysis for Detection and Identification of Bartonella Species

et al. 2001

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“…Bartonella bacilliformis is a Gram-negative, haemotrophic bacterium (Kreier & Ristic, 1981) that is capable of invading and replicating inside human erythrocytes and endothelial cells (Benson et al, 1986 ;McGinnis-Hill et al, 1992). The pathogen is transmitted to humans by the bite of a female sandfly (Lutzomyia spp.)…”

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A bacteriophage-like particle from Bartonella bacilliformis

Barbian

Minnick

2000

Microbiology

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Bartonella bacilliformis and Bartonella henselae, the respective agents of Oroya fever and cat-scratch disease in humans, are known to produce bacteriophage-like particles (BLPs) that package 14 kbp segments of the host chromosome. Data from this study suggest that other Bartonella species including Bartonella quintana, Bartonella doshiae and Bartonella grahamii also contain similar BLPs, as evidenced by the presence of a 14 kbp extrachromosomal DNA element in their genomes, whereas Bartonella elizabethae and Bartonella clarridgeiae do not. A purification scheme utilizing chloroform, DNase I and centrifugation was devised to isolate BLPs from B. bacilliformis. Intact BLPs were observed by transmission electron microscopy and were round to icosahedral in shape and approximately 80 nm in diameter. RFLP and Southern blot analysis of BLP DNA from B. bacilliformis suggest that packaging, while non-selective, is less than the near-random packaging previously reported for the B. henselae phage. Data also suggest that the linear, double-stranded BLP DNA molecules have blunt ends with noncovalently closed termini. Packaging of the BLP DNA molecules into a protein coat appears to be closely related to nucleic acid synthesis, as unpackaged phage DNA is not detectable within the host cell. SDS-PAGE analysis of purified BLPs from B. bacilliformis showed three major proteins with apparent molecular masses of 32, 34 and 36 kDa ; values that closely correspond to proteins found in B. henselae BLPs. Western blot analysis performed with patient convalescent serum showed that BLP proteins are slightly immunogenic in humans. To determine if BLPs contribute to horizontal gene transfer, mutants of B. bacilliformis were generated by allelic exchange with an internal fragment of the 16S-23S rDNA intergenic spacer region and a suicide vector construct, termed pKB1. BLPs from one of the resultant strains were able to package the mutagenized region containing the kanamycinresistance cassette ; however, numerous approaches and attempts at intraspecies transduction using these BLPs were unsuccessful.

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The 75-kilodalton antigen of Bartonella bacilliformis is a structural homolog of the cell division protein FtsZ

et al. 1997

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Bartonella bacilliformis is the etiologic agent of bartonellosis (Carrion's disease), a biphasic disease that is endemic among inhabitants of the western slopes of the Andes in Columbia, Ecuador, and Peru (11,14). The disease consists of a primary, acute, hematic phase characterized by fever, hemolytic anemia, and bacteremia, referred to as Oroya fever. This is followed by a secondary, chronic phase characterized by skin eruptions, referred to as verruga peruana. During the hematic phase, B. bacilliformis parasitizes almost 100% of the erythrocytes, resulting in their premature destruction, and leading to a very severe form of hemolytic anemia. In the secondary phase, B. bacilliformis invades endothelial cells, causing hemangiomas, possibly as a result of hyperproliferation of the terminal vasculature of the dermis and subcutaneous tissue (10). These hemangiomas bear a distinct clinical and histologic similarity to the hemangioma-like lesions of bacillary (epitheliod) angiomatosis (BA), which has now been associated with the newly described bacterium Bartonella henselae (previously, Rochalimaea henselae) (4,26,27). Interestingly, B. bacilliformis and B. henselae are also closely related phylogenetically, and it was suggested that these bacteria may have a common mechanism of pathogenesis (1).We were interested in isolating and characterizing immunodominant B. bacilliformis antigens because of their potential usefulness in diagnosis and in elucidating the pathogenesis of bartonellosis. Additionally, the presence of homologs of these antigens in B. henselae might be interesting from the standpoint of the pathological similarities between the lesions of verruga peruana and BA. To isolate immunodominant antigens of B. bacilliformis, we constructed a B. bacilliformis genomic DNA library and screened it with serum from a patient with the chronic verruga phase of bartonellosis. In this report, we describe the isolation and initial characterization of one of the clones expressing a 75-kDa antigen of B. bacilliformis. Based on the strong amino acid sequence homology with the cell division protein FtsZ, we suggest that the 75-kDa antigen is the B. bacilliformis homolog of FtsZ (FtsZ Bb ). This is the first study that shows that FtsZ proteins are immunogenic in humans. MATERIALS AND METHODSBacterial strains, plasmids, media, and serum. B. bacilliformis KC584 (ATCC 35686) and KC583 (ATCC 35685) were grown on heart infusion agar plates supplemented with 5% defibrinated rabbit blood (BBL) at 28°C for 6 to 8 days. Bacteria were harvested and resuspended in phosphate-buffered saline. The cells were then collected by centrifugation at 8,000 ϫ g, and the supernatant was discarded. The pellets were then resuspended in TE (10 mM Tris [pH 8.0], 1 mM EDTA) for DNA extraction.The XL-1Blue strain of Escherichia coli (Stratagene, Torrey Pines, La Jolla, Calif.) was grown on Luria broth (LB) with tetracycline (12.5 g/ml) or LB with ampicillin (50 g/ml) when transformed with pBluescript (Stratagene). E. coli SOLR cells were grown on LB wi...

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The Biology of Hemotrophic Bacteria

Cited by 39 publications

References 2 publications

Use of rpoB Gene Analysis for Detection and Identification of Bartonella Species

Use of rpoB Gene Analysis for Detection and Identification of Bartonella Species

A bacteriophage-like particle from Bartonella bacilliformis

The 75-kilodalton antigen of Bartonella bacilliformis is a structural homolog of the cell division protein FtsZ

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