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Culture-negative endocarditis due to Houston Complex Bartonella henselae acquired in Noumea, New Caledonia.

A 44-year-old man with a bioprosthetic aortic valve suffered destructive endocarditis with severe embolic disease due to Bartonella henselae infection. Multilocus sequence typing was successfully performed with crude preparations of operative tissue as templates, and the infecting organism was determined to be typical of the Houston clonal group, although it was never cultured from blood or tissue. This is the first report of B. henselae infection in the South Pacific, and it reminds one that B. henselae is a cause of potentially lethal culture-negative endocarditis which may respond poorly to conventional empirical therapy. Nothing is known of the epidemiology of the infection in this region, but it is likely to be common and to contain representatives of both major clonal complexes. This study emphasizes the ease with which multilocus sequence typing can be used directly with tissue, which is important because of suggestions of strain-dependent clinical outcomes.

Angiomatosis, Bacillary↗

Prevalence of Borrelia burgdorferi, Bartonella spp., Babesia microti, and Anaplasma phagocytophila in Ixodes scapularis ticks collected in Northern New Jersey.

PCR analysis of Ixodes scapularis ticks collected in New Jersey identified infections with Borrelia burgdorferi (33.6%), Babesia microti (8.4%), Anaplasma phagocytophila (1.9%), and Bartonella spp. (34.5%). The I. scapularis tick is a potential pathogen vector that can cause coinfection and contribute to the variety of clinical responses noted in some tick-borne disease patients.

Anaplasma phagocytophilum↗

Multispacer typing technique for sequence-based typing of Bartonella quintana.

Bartonella quintana is a worldwide fastidious bacterium of the Alphaproteobacteria responsible for bacillary angiomatosis, trench fever, chronic lymphadenopathy, and culture-negative endocarditis. The recent genome sequencing of a B. quintana isolate allowed us to propose a genome-wide sequence-based typing method. To ensure sequence discrimination based on highly polymorphic areas, we amplified and sequenced 34 spacers in a large collection of B. quintana isolates. Six of these exhibited polymorphisms and allowed the characterization of 4 genotypes. However, the strain variants suggested by the noncoding sequences did not correlate with the results of pulsed-field gel electrophoresis (PFGE), which suggested a higher degree of variability. Modification of the PFGE profile of one isolate after nine subcultures confirmed that rearrangement frequencies are high in this species, making PFGE unreliable for epidemiological purposes. The low extent of sequence heterogeneity in the species suggests a recent emergence of this bacterium as a human pathogen. Direct typing of natural samples allowed the identification of a fifth genotype in the DNA extracted from a human body louse collected in Burundi. We have named the typing technique herein described multispacer typing.

Animals↗

Diagnosis of cat scratch disease with detection of Bartonella henselae by PCR: a study of patients with lymph node enlargement.

Cat scratch disease (CSD) is mostly due to Bartonella henselae after inoculation of the organism through a skin injury. Since the causative bacteria cannot be easily cultured from human lymph node samples, the diagnosis usually relies on epidemiological, clinical, histological, and serological criteria (classical criteria). A study was performed to determine the diagnostic value of PCR analysis for the detection of B. henselae for the diagnosis of CSD and its place in the diagnostic strategy alongside the classical criteria. Over a 7-year period, lymph node biopsy specimens or cytopunctures from 70 patients were systematically tested by PCR for the presence of B. henselae DNA (htrA gene) in the Bacteriology Laboratory of the Hôpitaux Universitaires de Strasbourg. Serological testing by an immunofluorescence assay for B. henselae antibodies was also performed for each patient, and clinical, epidemiological, and histological data were collected. The patients were then divided into two groups according to the number of positive diagnostic criteria for CSD: 29 patients with definite CSD (two or more classical criteria) and 15 patients with possible CSD (less than two classical criteria). The remaining 26 patients for whom another diagnosis was retained were used as a control group. Among all criteria, PCR analysis had the best specificity (100%). The PCR assay for B. henselae was positive for 22 (76%; 95% confidence interval [CI95], 56.5 to 89.7%) of the 29 definite CSD patients and 3 (20%; CI95, 4.3 to 48.1%) of the 15 possible CSD patients. We then studied combinations of diagnostic criteria, including B. henselae PCR analysis. The best diagnostic performance was observed if at least two criteria were present among serologic, epidemiologic, histological, and molecular criteria.

Adolescent↗

Pseudoinfectious mononucleosis: a presentation of Bartonella henselae infection.

Six children presented during one year with clinical features of infectious mononucleosis, but with laboratory findings of leucocytosis with neutrophilia, increased erythrocyte sedimentation rate, and hypergammaglobulinaemia. Serology for Epstein-Barr virus, cytomegalovirus, adenovirus, and Toxoplasma gondii was negative, while anti-Bartonella henselae IgM with high IgG titre (>/=1/1024) was present in all. All children had contact with kittens. No specific treatment was administered and all recovered.

Bartonella henselae↗

Cloning, functional expression, and complementation analysis of an inorganic pyrophosphatase from Bartonella bacilliformis.

We have cloned the inorganic pyrophosphatase gene (ppa) from the facultative intracellular pathogen Bartonella bacilliformis and characterized its encoded product. The 531-bp gene is located approximately 1 kb downstream of, and in opposite orientation to, the invasion-associated locus (ialAB) of B. bacilliformis. The predicted protein encoded by ppa is 177 amino acid residues, which is in agreement with in vitro and in vivo synthesis of a protein with an apparent molecular mass of 22-23 kDa. The predicted B. bacilliformis pyrophosphatase (PPase) sequence is 53% identical and 85% similar to the E. coli PPase (EC 3.6.1.1), and contains all 12 of the amino acid residues implicated in the catalytic active site. The isolated B. bacilliformis PPase exhibits an activity of 51 +/- 2 mumol PO4 released/(mg protein.min) at 28 degrees C and pH 8, and is sensitive to inhibition by Ca2+. In keeping with other prokaryotic PPases, B. bacilliformis PPase activity occurs from pH 6 to 10 (optimal pH = 8) and demonstrates high thermostability in the presence of Mg2+ (highest activity at 55 degrees C, relative activity = 80 +/- 3% at pH 8). The cloned B. bacilliformis ppa is able to genetically complement a ppa- mutant strain of E. coli.

Amino Acid Sequence↗

The transcriptional response of human endothelial cells to infection with Bartonella henselae is dominated by genes controlling innate immune responses, cell cycle, and vascular remodelling.

The bacterial pathogen Bartonella henselae (Bh) is responsible for a broad range of clinical manifestations, including the formation of vascular tumours as the result of pathogen-triggered vasoproliferation. In vitro, the interaction of Bh with human umbilical vein endothelial cells (Huvec) involves (i) cytoskeletal rearrangements in conjunction with bacterial internalization, (ii) nuclear factor kappaB (NFkappaB)-dependent proinflammatory activation, (iii) the inhibition of apoptosis, and (iv) the modulation of angiogenic properties such as proliferation, migration, and tubular differentiation. To study the transcriptional signature of these pathogen-triggered changes of Huvec, we performed transcriptional profiling with Affymetrix U133 GeneChips. At 6 h or 30 h of infection, a total of 706 genes displayed a clear and statistically significant change of expression (>2.5-fold, t-test p-value<0.05). These included 314 up-regulated genes dominated by the innate immune response. The gene list comprises subsets of tumour necrosis factor alpha (TNFalpha, 99 genes) and interferon alpha (IFNalpha, 30 genes) inducible genes, which encode components of the NF-kappaB-dependent proinflammatory response and the type I IFN-dependent anti-infective response, respectively. The remaining set of 197 up-regulated genes mirrors other cellular changes induced by Bh, in particular proliferation and proangiogenic activation. The set of 362 down-regulated genes includes 41TNFalpha - or IFNalpha-suppressible genes, and 52 genes involved in cell cycle control or progression. This comprehensive analysis of Bh-triggered changes of the Huvec transcriptome identified candidate genes putatively involved in controlling innate immune responses, cell cycle, and vascular remodelling, and may thus provide the basis for functional studies of the molecular mechanisms underlying these pathogen-induced cellular processes.

Bartonella henselae↗

Disciforme keratitis caused by Bartonella henselae: an unusual ocular complication in cat scratch disease.

Cat scratch disease (CSD) is a common infectious disease, however, its association with disciforme keratitis is a previously unreported ocular complication. With the use of the 16S rDNA-PCR technique with subsequent DANN sequencing on corneal material obtained by corneal scrape we were able to identify Bartonella henselae in an unusual form of disciforme keratitis.

Aged↗

Role of dendritic cell-derived CXCL13 in the pathogenesis of Bartonella henselae B-rich granuloma.

Dendritic cells (DCs) initiate adaptive immunity and regulate the inflammatory response by producing inflammatory chemokines. This study was aimed to elucidate their role in the pathogenesis of the suppurative granuloma induced by Bartonella henselae infection, which characterizes cat scratch disease (CSD). In vitro DC infection by B. henselae results in internalization of bacteria, phenotypic maturation with increased expression of HLA-DR and CD86, and induction of CD83, CD208, and CCR7. In comparison to LPS-activated DCs, B henselae-infected DCs produce higher amounts of IL-10, whereas the production of IL-12p70 is reduced. Infected DCs also produce high levels of CXCL8 and CXCL13, 2 chemokines active respectively on neutrophils and B lymphocytes. These results provide the molecular basis for the morphogenesis of CSD granuloma, which typically contains high numbers of neutrophils and B cells. Remarkably, CSD granulomas in vivo contain CXCL13-producing DCs. We further demonstrate that the B cells in CSD granulomas are represented by monocytoid B cells and, worth noting, they express T-bet, a transcription factor able to induce a T-independent immunoglobulin (Ig) class switch in B lymphocytes. These findings suggest that the humoral immune response to B henselae initiates in the extrafollicular areas of infected lymph nodes and is regulated by DCs.

Animals↗

Isolation of Bartonella henselae from domestic cats in Japan.

During the period from January to March 1995, the authors first isolated Bartonella henselae from the blood of three (9.1%) of 33 domestic cats in Japan. The three cats were a 1.5-year male pet cat-old with urinary retention, and 6-year-old female pound and age-unknown female pet cats with no abnormalities. The blood was taken in a lysis-centrifugation tube (Wampole Isolator tube) and cultured on 5% rabbit-blood heart infusion agar plates at 35 degrees C in the 5% CO2 atmosphere. Visible tiny rough colonies developed 14 days after incubation. The isolates showed Gram-negative and pleomorphic rods in microscopic observation. The DNA extracted from the isolates was amplified by PCR using two primers, which were specific for the rikettsial citrate synthase gene. The isolates were identified as B. henselae from the patterns of digestion with TaqI and HhaI of the amplified gene. It was confirmed that cats in Japan harbored B. henselae in their blood, and that cats play a significant role as the reservoir of the organism.

Animals↗

Seroprevalence of Bartonella henselae and Toxoplasma gondii infections among pet cats in Kanagawa and Saitama Prefectures.

Seroprevalence of Bartonella henselae and Toxoplasma gondii was investigated among 471 pet cats obtained from seven private animal hospitals in Kanagawa and Saitama Prefectures during the period from May 1994 to June 1995. 'Furthermore, 67 randomly selected from the 471 serum samples were examined for the feline immunodeficiency virus (FIV) antibody and feline leukemia virus (FeLV) antigen. The antibody to B. henselae was examined by an indirect immunofluorescent antibody test. T. gondii, FIV and FeLV infections in cats were detected with respective commercial kits. Of the cat serum samples tested, 43 (9.1%) were found to be seropositive for B. henselae and 41 (8.7%) for T. gondii. The B. henselae-positive rate (12.9%) of male cats was significantly higher than that (5.2%) of female cats. On the other hand, T. gondii-positive rate was 9.1% in male and 8.7% in female cats and there was no significant difference in the positivity between sexes. The positive rate in each hospital varied from 0 to 19.5% for B. henselae and 4.9 to 18.8% for T. gondii. The ages of B. henselae- and T. gondii-positive cats were distributed from < 1-year-old to 14-year-old and the seropositivity increased with age of cats. Of the 67 cat serum samples, 16 and 6 cases were positive for FIV and FeLV, respectively. There was no relationship between these viral and B. henselae infections in cats.

Animals↗

Seroprevalence of Bartonella henselae and Toxoplasma gondii among healthy individuals in Thailand.

The seroprevalence of Bartonella henselae and Toxoplasma gondii among apparently healthy individuals, mainly blood donors, in Thailand was investigated by an indirect fluorescent antibody technique and by a latex agglutination test, respectively. Of 163 serum samples examined, 9 (5.5%) were found to be positive for B. henselae-IgG, 2 (1.2%) for B. henselae-IgM, and 5 (3.1%) for the T. gondii antibody. No significant difference was observed between male and female samples in the serological test with either B. henselae or T. gondii. The age of individuals with B. henselae-IgG was distributed from the 20s to the 70s, and B. henselae-IgM was found in the individuals of the 30s and 60s. The age of T. gondii positive samples ranged from the 20s to the 60s. In this study, the prevalence of B. henselae infection among healthy individuals in Thailand was serologically demonstrated for the first time.

Adolescent↗

Immune responses of immunocompetent and immunocompromised mice experimentally infected with Bartonella henselae.

The aim of this study is to understand host immune responses in immunocompetent and immunocompromised mice against Bartonella henselae infection. BALB/c and nude (BALB/c nu/nu) mice were inoculated intraperitoneally with 10(8) colony forming units of B. henselae (Houston-1 strain). Blood, brain, liver, spleen, kidney and bone marrow samples were collected 0, 3, 7, 14, 21 and 28 days after infection and submitted to bacteriological, serological and genetical examinations. B. henselae was isolated only from the liver 3 days after infection. DNA of the inoculums was detected by polymerase chain reaction from blood, liver, and spleen samples collected from BALB/c and blood from nude mice 3 and 7 days after infection. No bacterial DNA was detected from both BALB/c and nude mice thereafter during 4 weeks observation periods. These results indicate that the T-cell may not participate in the effective elimination of the organisms from mice. In addition, western blot analysis revealed that the antigens of 27.3- and 31.5-kDa reacted with IgM antibodies from the blood of BALB/c and nude mice after 3 days of infection, suggesting that these antigens were recognized by thymus-independent mechanism. Furthermore the antigens were detected from the culture-supernatants of B. henselae, indicating that these antigens were secreted from the organisms.

Angiomatosis, Bacillary↗

Parinaud syndrome caused by Bartonella henselae: case report.

The authors report a case of Parinaud syndrome (conjunctivitis with pre-auricular satellite adenitis) caused by Bartonella henselae, the etiologic agent of Cat Scratch Disease. The etiologic assessment of this case was performed by serum indirect immunofluorescence reaction and allowed for a better therapeutics and follow up, avoiding ineffective antibiotics and surgical interventions.

Animals↗

Studies on the growth of Bartonella henselae in the cat flea (Siphonaptera: Pulicidae).

Two out of three pools of cat fleas, Ctenocephalides felis (Bouche), that were fed Bartonella henselae-positive cat blood for 3 d and then bovine blood for 3 d, were polymerase chain reaction (PCR) positive for B. henselae. In a second experiment, three cats were inoculated with a streptomycin-resistant strain of B. henselae. After the cats were inoculated, caged cat fleas were fed on the cats during three different periods, and then pooled and transferred to noninfected recipient cats. In the first trial, the bacteria in the flea feces were below level of detection when the fleas were transferred from the infected cats to the recipient cat. After the fleas had fed on the recipient cat for 6 d, a bacteria level of 4.00 x 10(3) CFU/ mg was detected in the flea feces. Subsequently, the bacteria level increased for 4 d and then declined. In another experiment, the bacteria level in the flea feces was 1.80 x 10(3) CFU/mg at 2 h after collection and 3.33 x 10(2) CFU/mg at 72 h after collection. These data indicated that this strain of B. henselae can persist in flea feces in the environment for at least 3 d, and that B. henselae can multiply in the cat flea.

Animals↗

Seroprevalences of antibodies against Bartonella henselae and Toxoplasma gondii and fecal shedding of Cryptosporidium spp, Giardia spp, and Toxocara cati in feral and pet domestic cats.

OBJECTIVE: To compare seroprevalences of antibodies against Bartonella henselae and Toxoplasma gondii and fecal shedding of Cryptosporidium spp, Giardia spp, and Toxocara cati in feral and pet domestic cats. DESIGN: Prospective cross-sectional serologic and coprologic survey. ANIMALS: 100 feral cats and 76 pet domestic cats from Randolph County, NC. PROCEDURE: Blood and fecal samples were collected and tested. RESULTS: Percentages of feral cats seropositive for antibodies against B. henselae and T. gondii (93% and 63%, respectively) were significantly higher than percentages of pet cats (75% and 34%). Percentages of feral and pet cats with Cryptosporidium spp (7% of feral cats; 6% of pet cats), Giardia spp (6% of feral cats; 5% of pet cats), and T. cati ova (21% of feral cats; 18% of pet cats) in their feces were not significantly different between populations. Results of CBCs and serum biochemical analyses were not significantly different between feral and pet cats, except that feral cats had a significantly lower median PCV and significantly higher median neutrophil count. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that feral and pet cats had similar baseline health status, as reflected by results of hematologic and serum biochemical testing and similar prevalences of infection with Cryptosporidium spp, Giardia spp, and T. cati. Feral cats did have higher seroprevalences of antibodies against B. henselae and T. gondii than did pet cats, but this likely was related to greater exposure to vectors of these organisms.

Animals↗

Detection of Bartonella henselae by polymerase chain reaction in brain tissue of an immunocompromised patient with multiple enhancing lesions. Case report and review of the literature.

The authors report the first DNA-based diagnosis of Bartonella henselae cultured from a brain lesion in a patient with acquired immune deficiency syndrome. This human immunodeficiency virus-infected patient presented with altered mental status, fever, and diabetes insipidus. Magnetic resonance imaging revealed multifocal parenchymal and leptomeningeal involvement, which was confirmed on studies of tissue biopsy samples. Using the polymerase chain reaction and gene sequencing techniques, the authors definitively demonstrated the presence of B. henselae in the brain tissue biopsy specimen.

AIDS-Related Opportunistic Infections↗

Bartonella quintana in domestic cat.

We recovered Bartonella quintana DNA from dental pulp of a domestic cat. This study, the first to detect B. quintana in a nonhuman mammal, changes our understanding of the epidemiology of this infection and proposes that cats may be an emerging source of human infection.

Amino Acid Sequence↗