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Prevalence of Bartonella henselae, Bartonella clarridgeiae and the 16S rRNA gene types of Bartonella henselae among pet cats in Japan.

The authors investigated bacteriologically the prevalence of Bartonella infection among 690 pet cats derived from 10 private animal hospitals in six cities (Sapporo, Hokkaido Prefecture; Sendai, Miyagi Prefecture; Joetsu, Niigata Prefecture; Fujisawa, Kanagawa Prefecutre; Kyoto, Kyoto Prefecture; Sanda, Hyogo Prefecutre) and 4 counties (Mishima, Osaka Prefecture; Hikawa, Shimane Prefecture; Aira, Kagoshima Prefecture; Shimajiri, Okinawa Prefecture) located from the north to the south of Japan. Bartonella species were isolated from 7.2% (50/690) of all the cats examined. No Bartonella species were isolated from the cats in Sapporo or Sendai. The isolation rate varied from 2% in Joetsu and Sanda to 20% in Shimajiri. Bartonella clarridgeiae was isolated from two of 50 cats in Kyoto, three of 50 in Mishima and one of 50 in Shimajiri, but not in cats from the other cities or counties. Though the cats of Joetsu, Fujisawa, Kyoto, Sanda, Aira and Shimajiri were infected with either B. henselae or B. clarridgeiae, one of eight infected cats in Mishima was harboring both Bartonella species. Type I of 16S rRNA gene was the predominant type among the isolates of B. henselae, but only one isolate derived from Shimajiri was found to be of type II. Prevalence of B. clarridgeiae and the 16S rRNA gene type of B. henselae among cats in Japan was demonstrated for the first time in this investigation.

Animals↗

Evaluation of serological response to Bartonella henselae, Bartonella quintana and Afipia felis antigens in 64 patients with suspected cat-scratch disease.

The serological response to Bartonella henselae, B. quintana, and Afipia felis was assessed by an indirect fluorescence antibody test (IFAT) in 64 patients with suspected cat-scratch disease (CSD) recruited from the Bordeaux area in France. Blood samples were collected from 57 patients with chronic lymphadenopathy who underwent lymph-node biopsy with suggestive histopathologic features of CSD, and from an additional 7 patients with suspected CSD who underwent surgical incision and drainage because of lymph-node tenderness. Of the patients, 31 were male and 33 female, with a median age of 27 years (range 2-89). 69.8% reported cat and/or dog contact. Of the 26/64 (40.6%) patients, serum samples were positive at a titer of 1:100 or more for immunoglobulin G (IgG) antibodies (17 only to B. henselae, 1 only to B. quintana, 3 only to Afipia felis, and 5 to both B. henselae and B. quintana). IgM or IgA antibodies were also detected in 10 patients with IgG antibodies to B. henselae. 11 (17.2%) of the 64 patient serum samples were positive at a low titer of 1:50. These data suggested that serological response assessed by standard IFAT is not enough to confirm a CSD diagnosis.

Adolescent↗

Antibodies to Rickettsia rickettsii, Rickettsia typhi, Coxiella burnetii, Bartonella henselae, Bartonella quintana, and Ehrlichia chaffeensis among healthy population in Minas Gerais, Brazil.

Rickettsial diseases except those belonging to spotted fever group rickettsioses are poorly studied in South America particularly in Brazil where few epidemiological reports have been published. We describe a serosurvey for Rickettsia rickettsii, R. typhi, Coxiella burnetii, Bartonella henselae, B. quintana, and Ehrlichia chaffeensis in 437 healthy people from a Brazilian rural community. The serum samples were tested by indirected micro-immunoflourescence technique and a cutoff titer of 1:64 was used. The seroprevalence rates for R. rickettsii, R. typhi, C. burnetii, B. henselae, B. quintana, and E. chaffeensis were respectively 1.6% (7 samples); 1.1% (5 samples); 3.9% (17 samples); 13.7% (60 samples); 12.8% (56 samples), and 10.5% (46 samples). Frequent multiple/cross-reactivity was observed in this study. Age over 40 years old, urban profession, and rural residence were significantly associated with some but not all infections rate. Low seropositivity rates for R. rickettsii, R. typhi, and C. burnetii contrasted with higher rates of seropositivity for B. quintana, B. henselae, and E. chaffeensis. These results show that all tested rickettsial species or antigenically closely related possible exist in this particular region.

Adolescent↗

In vitro activities of telithromycin (HMR 3647) against Rickettsia rickettsii, Rickettsia conorii, Rickettsia africae, Rickettsia typhi, Rickettsia prowazekii, Coxiella burnetii, Bartonella henselae, Bartonella quintana, Bartonella bacilliformis, and Ehrlichia chaffeensis.

In vitro activities of telithromycin compared to those of erythromycin against Rickettsia spp., Bartonella spp., Coxiella burnetii, and Ehrlichia chaffeensis were determined. Telithromycin was more active than erythromycin against Rickettsia, Bartonella, and Coxiella burnetii, with MICs of 0.5 microg/ml, 0.003 to 0.015 microg/ml, and 1 microg/ml, respectively, but was inactive against Ehrlichia chaffeensis.

Anti-Bacterial Agents↗

[Seroprevalence of Bartonella henselae and Bartonella quintana infections in Poland in 1998-2001].

Bartonella henselae and Bartonella quintana infections result in illnesses with symptoms of severity ranging from mild lymphadenopathy (CSD) to systemic disease. The aim of the study was to estimate a prevalence of B. henselae and B. quintana infections in human in Poland. Serum samples collected from 265 patients in 1998-2001 were tested for the presence of antibodies specific to B. henselae and B. quintana. Levels of serum IgM and IgG antibodies to Bartonella henselae and Bartonella quintana were measured with indirect microimmunofluorescence test (MRL Diagnostic, USA). Cats' sera were assessed with indirect microimmunofluorescence test (MRL Diagnostic, USA) and goat immune serum anti-cat IgG FITC conjugate (Sigma, USA). Bartonella henselae specific antibodies were detected in 146 (57.0%) patients with lymphadenopathy. From that number 11.3% have shown specific Bartonella henselae IgM serum antibodies. Bartonella quintana infection was detected with serological methods in 4 patients. It has been found that CSD is a seasonal infection, with most cases occurring in autumn. Most cases of the disease have been recognized in children 8-16 years old. Most of CSD cases (30.1%) were detected in Mazowieckie voivodeship. There were no cases of CSD in Pomorskie, Podkarpackie, Lubuskie and Opolskie voivodeship. The seroprevalence of Bartonella sp. infections in cats was estimated on 86% (31/36). The highest titer of specific Bartonella henselae antibodies detected in cats was 1024. The number of detected Bartonella henselae infections in Poland is very low. It is very probable that the number of cases is underestimated in our country. Cat scratch disease is the most frequently clinically and serologically identified bartonellosis.

Adolescent↗

[Cat scratch disease caused by Bartonella henselae].

Bartonella henselae is an etiologic agent of cat-scratch disease and, in immunocompromised patients, of bacillary angiomatosis and other severe syndromes. Cat-scratch disease usually presents as lymphadenopathy, which resolves spontaneously within 2-4 months. The utility of antibiotic therapy remains controversial. In Tyrol four cases of human cat-scratch disease were diagnosed in children in 1994, yielding a prevalence of 0.7/100,000 per year. A 3-year-old boy had lymphadenitis coli since one year despite antituberculosis therapy which was initiated because of the histopathological picture and a positive tuberculin reaction (despite negative mycobacteria-cultures and -PCR). Two girls, age 9 and 13 years, had lymphadenitis at upper or lower extremities after cat-scratches from kittens. A 13-year-old boy presented with febrile illness and right hip pain, computer tomography revealed an osteolytic lesion; symptoms subsided within 3 weeks. Diagnosis of cat-scratch disease is based on cat contact, negative studies for other similar diseases, characteristic histopathologic features (if available), and results of an indirect immunofluorescence test (antigen: Houston-1 isolate, ATCC 49882). We believe that the availability of this serological test will increase the number of diagnosed cases of human Bartonella henselae infections.

Adolescent↗

Hemin-dependent growth and hemin binding of Bartonella henselae.

Bartonella henselae causes cat-scratch disease and bacillary angiomatosis peliosis. The bacteria reside in erythrocytes of asymptomatic cats, which represent the natural reservoir for this pathogen. B. henselae is usually grown on blood-enriched media. Growth experiments on Brucella medium without blood demonstrated that heme compounds are essential for the growth of B. henselae and can completely substitute the addition of blood components. The heme precursor protoporphyrin IX alone, or in combination with FeCl(2) or FeCl(3), as well as transferrin or lactoferrin did not support growth, indicating that B. henselae cannot synthesize heme itself. Hemin supported growth even when free iron was chelated, indicating that hemin is also used as an iron source. Binding assays showed that hemin starvation increased the binding capacity of B. henselae for hemin, providing evidence that the bacteria carry a specific hemin uptake system, which might be regulated by hemin.

Bartonella henselae↗

Phase variation in Bartonella henselae.

Bartonella henselae is a fastidious, Gram-negative bacterial pathogen of cats and humans. Previous workers have shown that serial passage in vitro leads to attenuation of virulence-associated attributes such as expression of pili, invasion of human epithelial cell lines and the stimulation of endothelial cell proliferation. In contrast to the published data, it was found that pilin expression is frequently preserved in organisms which have undergone phase variation in vitro. Transition from a slow-growing, dry agar-pitting (DAP) to a faster-growing, smooth non-agar-pitting (SNP) form appears to occur predictably and may reflect competition between two populations growing at different rates. Better survival of the slower-growing (DAP) form may explain its relatively easy retrieval from piliated SNP populations allowed to age on solid media. Pilin expression is associated with auto-agglutination in liquid suspension or broth cultures, and appears to be necessary but not sufficient for expression of the agar-pitting phenotype and for the formation of biofilms. Outer-membrane protein variation is seen in association with phase variation, but lipopolysaccharide expression is preserved in piliated as well as extensively passaged non-piliated isolates. The EagI/HhaI infrequent restriction site-PCR fingerprint, which has been previously used to discriminate between serotypes Marseille and Houston, is shown to alter with phase variation in vitro, and there is evidence that genetic change accompanies these events. The extent of genetic and phenotypic variability of phase-variant B. henselae has previously been underestimated. It may lead to new insights into the pathogenicity of this organism, and must be considered when interpreting data arising from such studies.

Animals↗

Determination of the rpoB gene sequences of Bartonella henselae and Bartonella quintana for phylogenic analysis.

Using the Genome Walker procedure, which allows PCR amplification of genomic DNA using a single gene-specific primer and direct automated sequencing methodology, we obtained the nucleotide sequence of the RNA polymerase beta subunit (rpoB) from Bartonella henselae and Bartonella quintana. A phylogenetic tree constructed from these data and other rpoB sequences available in GenBank is, in part, consistent with those previously derived from 16S rRNA gene sequences and confirms the position of Bartonella within the alpha subdivision of Proteobacteria. In fact, this analysis showed that rpoB data are similar to 16S rRNA data for the alpha, beta and gamma subdivisions of Proteobacteria. In contrast, concerning other bacteria included in our study, the topologies of phylogenetic trees were different. Based on the bootstrap values derived from rpoB phylogenic analysis, we believe that this molecule should contribute to better understanding the evolutionary process.

Amino Acid Sequence↗

Occurrence of Bartonella henselae and Bartonella quintana among human immunodeficiency virus-infected patients.

The purpose of this study was to determine the seroprevalence against Bartonella henselae and Bartonella quintana among a risk group, patients with HIV infection, and to identify the epidemiological factors involved. Our data indicate that the prevalence of Bartonella infection among HIV-infected patients is much greater than that in the healthy population of the same area and that Bartonella infection should be considered in the differential diagnosis for patients with HIV disease.

Angiomatosis, Bacillary↗

Proteomic analysis of the sarcosine-insoluble outer membrane fraction of the bacterial pathogen Bartonella henselae.

Bartonella henselae is an emerging zoonotic pathogen causing a wide range of disease manifestations in humans. In this study, we report on the analysis of the sarcosine-insoluble outer membrane fraction of B. henselae ATCC 49882 Houston-1 by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1-D SDS-PAGE) and two-dimensional nonequilibrium pH gradient polyacrylamide gel electrophoresis (2-D NEPHGE). Protein species were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and subsequent database query against the B. henselae genome sequence. Subcellular fractionation, application of the ionic detergent lauryl sarcosine, assessment of trypsin sensitivity, and heat modifiability of surface-exposed proteins represented valuable tools for the analysis of the outer membrane subproteome of B. henselae. 2-D NEPHGE was applied to display and catalogue a substantial number of proteins associated with the B. henselae sarcosine-insoluble outer membrane fraction, resulting in the establishment of a first 2-D reference map of this compartment. Thus, 53 distinct protein species associated with the outer membrane subproteome fraction were identified. This study provides novel insights into the membrane biology and the associated putative virulence factors of this pathogen of increasing medical importance.

Bacterial Proteins↗

Infection of fetal feline brain cells in culture with Bartonella henselae.

Bartonella henselae is known to cause central nervous system (CNS) disease in humans, and neurological signs have been observed in experimentally infected cats. However, the pathogenesis of CNS disease remains unclear. This study was undertaken to determine whether B. henselae infects feline fetal brain cells in vitro. Microglial-cell- and astrocyte-enriched cultures were inoculated with B. henselae. Giménez staining identified bacterial organisms within microglial cells by day 7 postinoculation. The viability of the intracellular bacteria was demonstrated by incubating cultures with gentamicin and plating cell lysate on agar. Electron microscopy identified intracellular organisms with characteristic Bartonella morphology but identified no ultrastructural abnormalities within infected microglial cells. No evidence of infection was seen in Bartonella-inoculated astrocyte cultures. These findings suggest a role for microglia in the pathogenesis of B. henselae-associated neurological disease.

Animals↗

The role of the host immune response in pathogenesis of Bartonella henselae.

Bartonella henselae can infect humans resulting in a wide range of disease syndromes including cat-scratch disease, fever with bacteremia, endocarditis, bacillary angiomatosis, and bacillary peliosis hepatis, among others. The nature and severity of the clinical presentation correlates well with the status of the hosts' immune system. Individuals with impaired immune function, including HIV infection, progress to systemic infections more often. Patients with intact immune function who become infected with B. henselae usually get cat-scratch disease, a disease that usually involves lymphadenopathy resulting from a strong cellular immune response to the bacterium. However, immunocompromised patients often progress to bacillary angiomatosis or bacillary peliosis hepatis. The reduced ability of the hosts immune response to control bacterial infection apparently results in a bacteremia of longer duration, and in some patients the presence of angiogenic lesions that are unique among bacterial infections to Bartonella. Recently, the role of immune effector cells that produce angiogenic cytokines upon stimulation with B. henselae has been proposed. Here, the current status of the role of the immune response in both controlling infection and in B. henselae-triggered immunopathogenesis is presented.

Animals↗

Seroprevalence of Bartonella henselae and Bartonella quintana infections in children from Central and Northern Jordan.

OBJECTIVE: To investigate the prevalence of antibodies to Bartonella henselae (B.henselae) and Bartonella quintana (B.quintana) among children from central and northern Jordan. METHODS: Sera from 482 children were randomly collected from referenced governmental hospitals in the central and northern parts of Jordan during the period between January 2001 to March 2003. An indirect immunofluorescent assay was used to determine serum antibody titers to B.henselae and B.quintana. Sera that were reactive at a dilution >/-1:64 were considered positive. In addition, blood from 20 cats belonged to children with high B.henselae titers were tested using polymerase chain reaction. RESULTS: Out of the 482 serum samples examined, 53 (11%) and 20 (4.1%) had positive antibody titers for B. henselae and B.quintana. Bartonella henselae was isolated from 4 cats that belonged to 3 children with high antibody titers to B.henselae-IgG. The seroprevalence of IgG antibodies to B.henselae was significantly higher (P<0.05) in children aged 7-10-years than in younger or older ones. Having a cat in the household and having a history of cat scratches or bites were strongly associated (P<0.01) with seropositivity to B.henselae-specific IgG. Cats ownership and history of cat scratches or bites had no impact on the prevalence of B.quintana. Seropositivity to B.henselae-specific IgG was significantly higher (P<0.01) in children from northern Jordan than in children from central Jordan. CONCLUSION: This study substantiates the presence of B.henselae in Jordan, documents the seroreactivity to 2 Bartonella antigens, and suggests that cat ownership and history of cat scratches or bites are important epidemiological risk factors for B.henselae infection in Jordan.

Animals↗

[Evaluation of isolation media for the detection of Bartonella henselae--isolation of Bartonella henselae from domestic cats].

Bartonella henselae is a causative agent of cat scratch disease. We preliminarily tested four media for the bacterial growth, including agar plates with sheep, horse or rabbit blood, and chocolate agar. Of these media, rabbit blood and chocolate agar plate were found to be more excellent for the growth than the medium with sheep or horse blood. Blood samples from 60 domestic cats in Yamaguchi Prefecture were then cultured using 7% rabbit blood agar plates and BACTEC9050 (BD), automated blood culture microbial detection system. B. henselae was isolated from six of the 60 (10%) blood samples. Tiny colonies of B. henselae were visible on the agar medium after one week of culture at 35 degrees C in the 5% CO2 atmosphere. BACTEC 9050 detected B. henselae in one of the 10 blood samples and it took two weeks to detect the bacteria automatically, though gram stain failed to show organisms in the blood culture bottle. In conclusion, rabbit blood or chocolate agar and incubation of agar media more than one week and of BACTEC more than two weeks are recommended for the detection of B. henselae.

Agar↗

Seroprevalence of and risk factors for Bartonella henselae and Bartonella quintana infections among pet cats in Jordan.

To determine the seroprevalences of Bartonella henselae and Bartonella quintana antibodies in Jordanian pet cats, serum samples from 153 cats from three geographical regions were analyzed. Seroprevalences were determined by indirect immunofluorescence. The true seroprevalences to B. henselae and B. quintana were 32 and 1.5%, respectively. The seroprevalence of B. henselae-specific antibodies in cats from northern Jordan was significantly higher than in cats from the central or southern parts of Jordan. The seroprevalence to B. henselae increased to age 2 years. Odds of seropositivity were higher in cats living outdoors, showing hunting behavior, having a flea infestation and of a mixed breed. No association was detected with sex.

Animals↗

Heat shock response and groEL sequence of Bartonella henselae and Bartonella quintana.

Transmission of Bartonella species from ectoparasites to the mammalian host involves adaptation to thermal and other forms of stress. In order to better understand this process, the heat shock response of Bartonella henselae and Bartonella quintana was studied. Cellular proteins synthesized after shift to higher temperatures were intrinsically labelled with [25S]methionine and analysed by gel electrophoresis and fluorography. The apparent molecular masses of three of the major heat shock proteins produced by the two Bartonella species were virtually identical, migrating at 70, 60 and 10 kDa. A fourth major heat shock protein was larger in B. quintana (20 kDa) than in B. henselae (17 kDa). The maximum heat shock response in B. quintana and B. henselae was observed at 39 degrees C and 42 degrees C, respectively. The groEL genes of both Bartonella species were amplified, sequenced and compared to other known groEL genes. The phylogenetic tree based on the groEL alignment places B. quintana and B. henselae in a monophyletic group with Bartonella bacilliformis. The deduced amino acid sequences of Bartonella GroEL homologues contain signature sequences that are uniquely shared by members of the Gram-negative alpha-purple subdivision of bacteria, which live within eukaryotic cells. Recombinant His6-GroEL fusion proteins were expressed in Escherichia coli to generate specific rabbit antisera. The GroEL antisera were used to confirm the identity of the 60 kDa Bartonella heat shock protein. These studies provide a foundation for evaluating the role of the heat shock response in the pathogenesis of Bartonella infection.

Amino Acid Sequence↗

Efficacy of enrofloxacin or doxycycline for treatment of Bartonella henselae or Bartonella clarridgeiae infection in cats.

Enrofloxacin and doxycycline are antimicrobial agents used to treat bacterial diseases of cats. In vitro susceptibility data indicate that either drug should be effective against Bartonella species. In vivo efficacies of these drugs for eradication of chronic Bartonella henselae or Bartonella clarridgeiae infections were examined in 18 experimentally infected cats and 25 naturally exposed cats treated with enrofloxacin (22.7 mg given orally [PO] every 12 h [q12h] [14 days, n = 10; 28 days, n = 13]) or with doxycycline (25 mg PO q12h [14 days, n = 9; 28 days, n = 8]) or not treated (n = 3). Plasma drug concentrations were determined in experimental cats by high-performance liquid chromatography. Only 23 of 43 cats enrolled ultimately met inclusion criteria. Bacteremia was eliminated for 12 to 25 weeks posttreatment in four of seven cats receiving 14 days of enrofloxacin, five of seven cats receiving 28 days of enrofloxacin, one of six cats receiving 14 days of doxycycline, and one of two cats receiving 28 days of doxycycline. Defining a negative result by blood culture as treatment success may be erroneous; these results may reflect the insensitivity of blood culture or the relapsing nature of Bartonella bacteremia. Our results suggest that MICs obtained with axenic media do not predict antimicrobial activity against intracellular Bartonella, that a long treatment course is required to eliminate infection, and that duration of therapy correlates with pretreatment bacterial load. Given current concern about the development of antimicrobial resistance, we would reserve recommendation for treatment to cats owned by an immunocompromised individual or as an alternative to euthanasia of a pet.

Administration, Oral↗