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PubMed2001

Molecular characterization of first human Bartonella strain isolated in Italy.

The aim of this study was to characterize a Bartonella strain (BA-1) isolated from a blood culture of an Italian, human immunodeficiency virus-positive patient with bacillary angiomatosis. We analyzed the isolate using molecular biology methods such as whole-cell fatty acid analysis, PCR-restriction fragment length polymorphism analysis, type-specific 16S rRNA PCRs, sequence analysis of the 16S rRNA, pulsed-field gel electrophoresis, and arbitrarily primed PCR. The BA-1 isolate turned out to be a Bartonella quintana strain, similar but not identical to B. quintana Oklahoma, which was used as a control strain.

Adult↗
PubMed2001

Genetic variability and prevalence of Bartonella henselae in cats in Berlin, Germany, and analysis of its genetic relatedness to a strain from Berlin that is pathogenic for humans.

Nineteen Bartonella henselae strains and one Bartonella clarridgeiae strain were isolated from blood samples of 97 pet cats and 96 stray cats from Berlin, Germany, indicating prevalence rates of 1 and 18.7%, respectively, for B. henselae and 0 and 1%, respectively, for B. clarridgeiae. Eighteen of 19 B. henselae isolates corresponded to 16S rRNA type II. Pulsed-field gel electrophoresis (PFGE) analysis revealed seven different PFGE types among the feline B. henselae strains. Interestingly, all feline isolates displayed low genetic relatedness to B. henselae strain Berlin-1, which is pathogenic for humans.

Animals↗
PubMed2002

Limited diversity among human isolates of Bartonella henselae.

A study of 59 isolates of Bartonella henselae reveals relatively limited diversity among those of human origin (n = 28). Either of two distinct alleles of both gltA and 16S ribosomal DNA (rDNA) was found in all isolates, with a high level of congruity between 16S and gltA inheritance among proven human pathogens. Human isolates from all over Eastern Australia were most commonly 16S rDNA (Bergmans) type I, with the same gltA allele as the type strain (Houston-1). Comparable feline isolates were more commonly 16S type II, with less congruity of inheritance between 16S and gltA alleles. Previously described arbitrarily primed PCR and EagI-HhaI infrequent restriction site PCR fingerprinting techniques separated Bartonella species effectively but lacked discriminating power within B. henselae. Examination of the 16-23S intergenic spacer region revealed for several strains several point mutations as well as a repeat sequence of unknown significance which is readily detected by HaeIII restriction fragment length polymorphism analysis. The bacteriophage-associated papA gene was present in all isolates. Enterobacterial repetitive intergenic consensus PCR proved to be a useful and robust typing tool and clearly separated human isolates (including imported strains) from the majority of feline isolates. Our data are consistent with published evidence and with previous suggestions of intragenomic rearrangements in the type strain and suggest that human isolates come from a limited subset of B. henselae strains. They strengthen arguments for careful exploration of genotype-phenotype relationships and for the development of a multilocus enzyme electrophoresis and multilocus sequence typing-based approach to the phylogeny of B. henselae.

Animals↗
PubMed2002

Genotypic characteristics of two serotypes of Bartonella henselae.

The study of 16S rRNA gene sequences of all isolates of Bartonella henselae obtained in our laboratory and others from human patients or cats has revealed two genotypes according to the sequence of the 16S rRNA gene. Two isolates of these genotypes have previously been related to two different serotypes, and lack of cross-protection of the two serotypes has been demonstrated in cats. We investigated the grouping of eight strains of B. henselae on the basis of 16S ribosomal DNA, 35-kDa protein, Pap 31 protein, and internal transcribed spacer (ITS) gene sequencing; sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles; and monoclonal antibody reactivity studies. Houston-1, 90-615, and SA2 strains showed the same patterns in SDS-PAGE, but they differed from the patterns of B. henselae isolates URBHLLY8, URBHLIE9, Cat6, Fizz, and CAL-1. Nine monoclonal antibodies derived from BALB/c mice immunized with B. henselae Houston-1 strain reacted only with strains Houston-1, 90-615, and SA2, and not with any other Bartonella strains. The two serogroups corresponded with two genotypes based on differences in the sequences of the genes encoding 16S rRNA, 35-kDa protein, and Pap 31 protein. Sequences of ITS genes were highly divergent among strains, as each had a unique sequence and the subdivision was not supported by DNA-DNA relatedness study. Study of 22 additional strains of B. henselae isolated from French bacteremic cats demonstrated that they all belong to one or the other of the proposed serotype or genotype.

Angiomatosis, Bacillary↗
PubMed2003

Characterization of the natural population of Bartonella henselae by multilocus sequence typing.

Investigations of the population genetics of Bartonella henselae have demonstrated a high level of diversity among strains, and the delineation of isolates into one of two subtypes, type I (Houston) and type II (Marseille), represented by specific 16S ribosomal DNA (rDNA) sequences, has long been considered the most significant genotypic division within the species. This belief is challenged by recent work suggesting a role for horizontal gene exchange in generating intraspecies diversity. We attempted to resolve this issue and extend exploration of the population structure of B. henselae by using multilocus sequence typing (MLST) to examine the distribution of polymorphisms within nine different genes in a sample of 37 human and feline isolates. MLST distinguished seven sequence types (STs) that resolved into three distinct lineages, suggesting a clonal population structure for the species, and support for these divisions was obtained by macrorestriction analysis using pulsed-field gel electrophoresis. The distribution of STs among isolates recovered from human infections was not random, and such isolates were significantly more often associated with one particular ST, lending further support to the suggestion that specific genotypes contribute disproportionately to the disease burden in humans. All but one isolate lay on lineages that bore the representative strain of either the Houston or Marseille subtype. However, the distribution of the two 16S rDNA alleles among the isolates was not entirely congruent with their lineage allocations, indicating that this is not a sensitive marker of the clonal divisions within the species. The inheritances of several of the genes studied could not be reconciled with one another, providing further evidence of horizontal gene transfer among B. henselae strains and suggesting that recombination has a role in shaping the genetic character of bartonellae.

Animals↗
PubMed2004

Evaluation of indirect fluorescence antibody assay for detection of Bartonella clarridgeiae and Seroprevalence of B. clarridgeiae among patients with suspected cat scratch disease.

The possibility of Bartonella clarridgeiae being a causative agent of cat scratch disease (CSD) was investigated by using indirect fluorescence antibody assays with 288 suspected CSD patients. Immunoglobulin G antibody to noncocultivated B. clarridgeiae was suitable only for detection of B. clarridgeiae antibody. Significant cross-reactivity between Bartonella henselae and B. clarridgeiae was noted, and no CSD case caused by B. clarridgeiae was detected.

Antibodies, Bacterial↗
PubMed2005

Bartonella species as a potential cause of epistaxis in dogs.

Infection with a Bartonella species was implicated in three cases of epistaxis in dogs, based upon isolation, serology, or PCR amplification. These cases, in conjunction with previously published reports, support a potential role for Bartonella spp. as a cause of epistaxis in dogs and potentially in other animals, including humans.

Animals↗
PubMed1994

Transformation of Bartonella bacilliformis by electroporation.

Bartonella bacilliformis is a member of the order Rickettsiales, family Bartonellaceae. The bacterium is an intracellular parasite of human erythrocytes. To date, members of the family Bartonellaceae have not been transformed by standard chemical methods. We report the first successful transformation of a member of the Bartonellaceae family, B. bacilliformis, by the method of electroporation. The optimal conditions for electroporation of B. bacilliformis include a field strength of 12.5 kV/cm and a time constant of 5 ms using 0.2-cm cuvettes. With these parameters and the cosmid pEST (RK2 replicon), a transformation efficiency of 7.8 x 10(5) was obtained. Transformants were readily cultured on medium containing kanamycin sulfate at concentrations ranging from 15 to 600 micrograms/mL. Bacterial survival was approximately 31% under optimal electroporation conditions, and the maximal number of transformants was obtained with 80 ng of pEST DNA. Bartonella bacilliformis was verified as the transformed organism by a comparison of transformant protein profiles with those of wild-type B. bacilliformis using sodium dodecyl sulfate polyacrylamide gel electrophoresis, and detection of the exogenous plasmid in DNA from the transformed bacteria by DNA hybridization. Transformations using the plasmids pMK20, pML31, and pUCK18 (containing the replicons ColE1, F, and pMB1, respectively) were not successful.

Bartonella↗
PubMed2005

Isolation of bacteriophages from Bartonella vinsonii subsp. berkhoffii and the characterization of Pap31 gene sequences from bacterial and phage DNA.

Bacteriophages enhance bacterial survival, facilitate bacterial adaptation to new environmental conditions, assist in the adaptation to a new host species, and enhance bacterial evasion or inactivation of host defense mechanisms. We describe the detection and purification of a novel tailed bacteriophage from Bartonella vinsonii subsp. berkhoffii, which was previously described as a bacteriophage-negative species. We also compare B. vinsonii subsp. berkhoffi Pap31 bacteriophage gene sequences to B. henselae (Houston I), and B. quintana (Fuller) bacteriophage Pap31 sequences. Negative staining electron microscopy of log phase culturesof B. vinsonii subsp. berkhoffii identified bacteriophages, possessing a 50-nm icosahedric head diameter and a 60- to 80-nm contractile tail. Sequence analysis of the bacteriophage Pap31 gene from B. vinsonii subsp. berkhoffii showed three consensus sequences and a 12-bp insertion when compared with Pap31 gene sequences from B. henselae (Houston I) and B. quintana (Fuller) bacteriophages. Isolation of B. vinsonii subsp. berkhoffii bacteriophages containing a Pap31 gene suggests that this heme-binding protein gene might play an important role in bacterial virulence through the genetic exchange of DNA within this subspecies. Defining phage-associated genes may also contribute to the enhanced understanding of the evolutionary relationships among members of the genus Bartonella.

Bacteriophages↗
PubMed2005

Infection of human CD34+ progenitor cells with Bartonella henselae results in intraerythrocytic presence of B. henselae.

Although there is evidence that endothelial cells are important targets for human pathogenic Bartonella species, the primary niche of infection is unknown. Here we elucidated whether human CD34+ hematopoietic progenitor cells (HPCs) internalize B. henselae and may serve as a potential niche of the pathogen. We showed that B. henselae does not adhere to or invade human erythrocytes. In contrast, B. henselae invades and persists in HPCs as shown by gentamicin protection assays, confocal laser scanning microscopy (CLSM), and electron microscopy (EM). Fluorescence-activated cell sorting (FACS) analysis of glycophorin A expression revealed that erythroid differentiation of HPCs was unaffected following infection with B. henselae. The number of intracellular B. henselae continuously increased over a 13-day period. When HPCs were infected with B. henselae immediately after isolation, intracellular bacteria were subsequently detectable in differentiated erythroid cells on day 9 and day 13 after infection, as shown by CLSM, EM, and FACS analysis. Our data provide, for the first time, evidence that a bacterial pathogen is able to infect and persist in differentiating HPCs, and suggest that HPCs might serve as a potential primary niche in Bartonella infections.

Angiomatosis, Bacillary↗
PubMed2005

Bacillary angiomatosis caused by Bartonella quintana.

Bacillary angiomatosis is a disorder of neovascular proliferation involving skin and lymph nodes of immunosuppressed patients. Bartonella henselae or Bartonella quintana have been inculpated as causative by direct culture or PCR amplification of DNA sequences. Here, we report the clinical evolution of a patient infected with human immunodeficiency virus (HIV) whose B. quintana infection was diagnosed by PCR.

Angiomatosis, Bacillary↗
PubMed1997

Interaction of Bartonella henselae with endothelial cells results in bacterial aggregation on the cell surface and the subsequent engulfment and internalisation of the bacterial aggregate by a unique structure, the invasome.

Vascular colonisation by Bartonella henselae may cause vaso-proliferative tumour growth with clumps of bacteria found in close association with proliferating endothelial cells. By using B. henselae-infected human umbilical vein endothelial cells as an in vitro model for endothelial colonisation, we report here on a novel mechanism of cellular invasion by bacteria. First, the leading lamella of endothelial cells establishes cellular contact to sedimented bacteria and mediates bacterial aggregation by rearward transport on the cell surface. Subsequently, the formed bacterial aggregate is engulfed and internalised by a unique host cellular structure, the invasome. Completion of this sequence of events requires 24 hours. Cortical F-actin, intercellular adhesion molecule-1 and phosphotyrosine are highly enriched in the membrane protrusions entrapping the bacterial aggregate. Actin stress fibres, which are anchored to the numerous focal adhesion plaques associated with the invasome structure, are typically found to be twisted around its basal part. The formation of invasomes was found to be inhibited by cytochalasin D but virtually unaffected by nocodazole, colchicine or taxol, indicating that invasome-mediated invasion is an actin-dependent and microtubuli-independent process. Bacterial internalisation via the invasome was consistently observed with several clinical isolates of B. henselae, while a spontaneous mutant obtained from one of these isolates was impaired in invasome-mediated invasion. Instead, this mutant showed increased uptake of bacteria into perinuclear localising phagosomes, suggesting that invasome-formation may interfere with this alternative mechanism of bacterial internalisation. Internalisation via the invasome represents a novel paradigm for the invasion of bacteria into host cells which may serve as a cellular colonisation mechanism in vivo, e.g. on proliferating and migrating endothelial cells during Bartonella-induced vaso-proliferative tumour growth.

Actins↗
PubMed2000

Three cases of cat scratch disease diagnosed by indirect immunofluorescence antibody assay and/or polymerase chain reaction of 16S rRNA gene of Bartonella henselae.

Three suspected cases of cat scratch disease were diagnosed by indirect immunofluorescence antibody assay and/or polymerase chain reaction. Patient 1 was a 10-year-old female who presented swelling of the right axillary [corrected] lymph nodes with pain and fever. She kept a kitten, and many scratches were observed on her both legs and dorsum manus. Antibody titers against Bartonella (B.) henselae were 1:32 for IgM 3 weeks after the onset of the symptoms and 1:64 for IgG 8 weeks after the onset. The DNA for 16S rRNA type I of B. henselae was detected from the blood sample obtained 3 weeks after the onset of symptoms by polymerase chain reaction for the first time in Japan. Patient 2 was a 22-year-old female veterinary student with a cat scratch at the bottom of her neck by a male kitten. She developed a papule at the scratch, slight fever, and neck pain. Although both Bartonella-specific IgG and IgM antibodies were negative before the scratch, the IgG antibody titer rose to 1:512 14 weeks after the onset. B. henselae was isolated from the kitten and its DNA found to be for 16S rRNA type I by PCR. Patient 3 was a 23-year-old female veterinary student with a cat scratch on her left forearm. A small reddish papule developed on the scratch, and she experienced swelling of the left axillary [corrected] lymph node and pain. Both the IgG and IgM antibodies against B. henselae were negative before the cat scratch, and the IgG titer rose significantly to 1:128 and 1:1,024 in 2 and 5 weeks, respectively, after the onset of the symptoms.

Adult↗
PubMed2002

Detection of Bartonella quintana from body lice (Anoplura: Pediculidae) infesting homeless people in Tokyo by molecular technique.

We report detection of Bartonella quintana Brenner, the pathogenic agent of trench fever, from body lice, Pediculus humanus L., infesting homeless people in Tokyo by polymerase chain reaction. Two of 12 (16.7%) homeless were infested with Bartonella-positive body lice. From the current status of the recent increase of homeless people in many large cities of the developed countries, a medical examination of homeless people should be carefully performed in the consideration of trench fever. Sampling of body lice from clothing of homeless people is recommended for quick and accurate diagnosis of trench fever through the detection of B. quintana DNA.

Animals↗
PubMed2006

[Bartonella henselae as a cause of optical nerve neuritis].

INTRODUCTION: Bartonella henselae is included into the group of gram-negative bacteria that can cause not so rare disease known as cat-scratch disease (CSD). This disease is characterized by the specific general symptoms, and the complications in the eyes can be manifested in the form of neuroretinitis, follicular conjunctivitis and focal chorioretinitis. In this paper clinical and epidemiological characteristics of a patient with ophthalmologic complications caused by Bartonella henselae are described. We indicate diagnostic possibilities and the criteria for making the diagnosis. CASE REPORT: We presented a 42-year-old female patient with CSD symptoms, and with a registered neuritis as an ophthalmologic complication. Two weeks after the occurrence of a scratch on the hand after the contact with a cat, there was a slight lymphadenopathy of the axial area of the left side, accompanied by light febrility and weakness. A week after these symptoms occuring, the patient complained of a reduction of the visual acuity in her left eye. The established visual acuity in the left eye was 0.1 with ophthalmological findings of a light edema of the optical disk and a partial star-like edema in the nasal half of the macula. Central scotoma was present, with the extension of the blind spot and the constriction of central isopter of the left eye visual field. Fluorescein biomicroscopy revealed an intensive leaking of fluoresceins at the level of optical disk and macular region in an early arterial phase, without the appearance of pathological phenomena at the level of retinal blood vessels. With the application of doxycyline 100 mg two times a day, and systemic application of prednisolone (at the initial doses of 120 mg), after a two-week period, there was a full recovery of the visual acuity, out the optical disk edema, and the presence of light edema in the left eye macula receded. The complete disappearance of the edema in the macula was registered four weeks following the application of the therapy. CONCLUSION: Cat-scratch disease can be recognized by means of characteristic general symptoms, and it must be considered in persons with ophthalmologic picture of neuroretinitis. In spite of the good prognosis for the general condition of a patient, it is advised to apply antibiotic therapy in the cases where ophthalmologic complications appear. We recommend the application of doxycyline 100 mg two times a day, for a month. When pronounced edema of the optical disk and edema in the macula are present, a systematic application of corticosteroids is necessary.

Adult↗
PubMed2005

Clinicopathologic findings in dogs seroreactive to Bartonella henselae antigens.

OBJECTIVE: To assess the potential clinical relevance of seroreactivity to Bartonella henselae antigens in dogs. ANIMALS: 40 dogs seroreactive to B henselae and 45 dogs that did not seroreact to B henselae. PROCEDURE: A case-control study was conducted. Clinical and clinicopathologic findings were extracted from medical records of each dog. RESULTS: Statistical differences were not detected between dogs seroreactive or nonseroreactive to B henselae when analyzed on the basis of disease category or results of hematologic, biochemical, urine, or cytologic analysis. However, seroreactivity to B henselae antigens was detected in 2 of 4 dogs with a clinical diagnosis of granulomatous meningoencephalitis, 3 of 4 dogs with immune-mediated hemolytic anemia, 3 of 4 dogs with infective endocarditis, 2 of 3 dogs with lymphoid neoplasia, and 5 of 10 dogs with polyarthritis. Additionally, seroreactivity to B henselae antigens was detected in 18 of 34 thrombocytopenic dogs and 14 of 27 dogs with neutrophilia. CONCLUSIONS AND CLINICAL RELEVANCE: Significant associations were not detected between seroreactivity to B henselae and various diseases. Prospective epidemiologic studies investigating specific diseases, such as meningoencephalitis or polyarthritis, and specific hematologic abnormalities, such as immunemediated hemolytic anemia or thrombocytopenia, should be conducted to further define the potential clinical relevance of antibodies against B henselae in dogs. IMPACT FOR HUMAN MEDICINE: Bartonella organisms are increasingly reported as pathogens that induce are increasingly reported as pathogens that induce chronic infections in humans and dogs. Dogs may serve as natural candidates for future study of the disease in humans.

Animals↗
PubMed2003

Rickettsial, ehrlichial and Bartonella infections of the myocardium and pericardium.

Myocarditis and pericarditis are uncommon complications of human rickettsial, ehrlichial and Bartonella infections. Myocardial inflammation usually occurs in the setting of acute disseminated infection. Organisms associated with myocarditis include: Rickettsia rickettsii, R. conorii, Orientia tsutsugamushi, Coxiella burnetii, Anaplasma phagocytophila (the causative agent of Human Granulocytic Ehrlichiosis) and Bartonella henselae. Pericarditis has been described in the setting of R. conorii and Coxiella burnetii infections. This article reviews the epidemiology, pathologic characteristics, clinical manifestations, diagnosis and treatment of myocarditis and pericarditis caused by these organisms.

Animals↗