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Bartonella koehlerae, a new cat-associated agent of culture-negative human endocarditis.

Bartonella koehlerae is reported for the first time to be a human pathogen that causes culture-negative endocarditis. It is also shown that this species, isolated twice before from domestic cats, can be recovered as well from a stray cat population in Israel. This work follows a recent report of the same case in which the causative agent was misidentified as B. henselae, based on serology and PCR-restriction fragment length polymorphism (RFLP) analysis (A. Schattner, O. Zimhony, B. Avidor, and M. Gilad, Lancet 361:1786, 2003). B. koehlerae was identified in the valvular tissue of an endocarditis patient by DNA sequencing of the PCR products of two Bartonella genes: the genes for citrate synthase (gltA) and riboflavin synthase (ribC). The commonly used PCR-RFLP analysis of the TaqI-digested gltA PCR product did not distinguish between B. koehlerae and B. quintana or between B. elizabethae and B. clarridgeiae. PmlI digestion of the gltA amplification product failed to differentiate between B. quintana, B. clarridgeiae, and B. elizabethae. RFLP analysis of the heat shock protein (htrA) gene by TaqI digestion misidentified B. koehlerae as B. henselae. However, RFLP analysis of the ribC PCR product, digested with TaqI, was able to distinguish between the human endocarditis-associated Bartonella species tested, B. henselae, B. quintana, B. elizabethae, and B. koehlerae, as well as between the cat-associated Bartonella species, B. henselae and B. clarridgeiae. Given the expanding number of Bartonella species emerging as human pathogens, it is suggested that PCR-RFLP analysis for the diagnosis of Bartonella infections target several genes and be coupled with DNA sequencing to avoid species identification.

Bartonella↗

Bartonella henselae infection presenting as a unilateral panuveitis simulating Vogt-Koyanagi-Harada syndrome.

PURPOSE: To report an unusual ocular manifestation of cat scratch disease. DESIGN: Observational case report. METHODS: Review of the clinical, laboratory, photographic, and angiographic records of a patient with cat scratch disease. RESULTS: A 54-year-old woman presented with counting fingers visual acuity in the right eye associated with optic disk edema, diffuse choroidal thickening, and panuveitis. Fluorescein angiography showed disk leakage and hyperfluorescent spots with late leakage suggestive of Vogt-Koyanagi-Harada disease. She was diagnosed with cat scratch disease by serum antibody titers and clinical course. CONCLUSIONS: Ocular manifestations of cat scratch disease can include diffuse thickening of the choroid. Cat scratch disease may manifest with angiographic features suggestive of Vogt-Koyanagi-Harada disease.

Anti-Bacterial Agents↗

An update on human bartonelloses.

The data available on Bartonella species has increased rapidly in the last decade with 12 new species and subspecies being described and bartonellas being found to be associated with an increasing spectrum of clinical syndromes in immunocompetent and immunocompromised patients. In addition B. weissii is found in cats. This rapid expansion of knowledge has come about with the development of new culture and molecular biology techniques to isolate and identify the organisms. Much of the information on the bartonellas and the emerging infectious diseases they cause has been derived from the USA and Europe although there is growing evidence that the organisms may be important pathogens in other regions including southern Africa. In this review we describe the bacteriology, epidemiology, clinical and laboratory signs, diagnosis and treatment of bartonella infections and the most recent data on bartonellas in southern Africa.

Africa, Southern↗

[Disciform keratatis caused by Bartonella henselae infection: detection of a rare ocular complication of cat-scratch disease with PCR].

BACKGROUND: Cat scratch disease (CSD) is a common infectious disease, however its association with a disciforme keratitis is to our knowledge a previously unreported ocular complication. PATIENT: A 65-year old female was treated unsuccessfully with topical antibiotics, acyclovir and corticosteroids for several weeks because of a disciforme keratitis. Repeatedly, microbiological investigations (microscopy and culture) were negative. 16S rDNA-PCR with subsequent DNA-sequencing on material obtained by corneal scrape identified Bartonella henselae. Based on this result we have initiated a treatment with topical cephotaxim and within a short period of time the cornea healed with a paracentral scar. CONCLUSION: 16S rDNA-PCR was very helpful in identifying the intracellular sited bacterium Bartonella henselae as the causative pathogen of an unusual form of disciforme keratitis.

Aged↗

Bartonella henselae in Ixodes ricinus ticks (Acari: Ixodida) removed from humans, Belluno province, Italy.

The potential role of ticks as vectors of Bartonella species has recently been suggested. In this study, we investigated the presence of Bartonella species in 271 ticks removed from humans in Belluno Province, Italy. By using primers derived from the 60-kDa heat shock protein gene sequences, Bartonella DNA was amplified and sequenced from four Ixodes ricinus ticks (1.48%). To confirm this finding, we performed amplification and partial sequencing of the pap31 protein and the cell division protein ftsZ encoding genes. This process allowed us to definitively identify B. henselae (genotype Houston-1) DNA in the four ticks. Detection of B. henselae in these ticks might represent a highly sensitive form of xenodiagnosis. B. henselae is the first human-infecting Bartonella identified from Ixodes ricinus, a common European tick and the vector of various tickborne pathogens. The role of ticks in the transmission of bartonellosis should be further investigated.

Animals↗

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↗

Culture-positive Bartonella quintana endocarditis.

A 50-year-old alcoholic man with a 1-year history of pyrexia of unknown origin was admitted with symptoms of endocarditis. Large vegetations on his mitral and aortic valve were found on echocardiography. Using the Bactec 9240 system. Bartonella quintana could be grown from two culture sets of blood collected before the start of antimicrobial therapy. This is the first case report of culture-proven bartonella infection in the UK. and is noteworthy because Bartonella quintana was detected in conventional blood culture systems after prolonged incubation.

Bacteremia↗

Mycoplasma and Bartonella in cats from the tropical tourist Gili Islands, Indonesia.

Bartonella spp. and haemotropic Mycoplasma spp. are important vector-borne bacteria of veterinary and zoonotic relevance, yet information on their circulation in Indonesian island ecosystems remains limited. We investigated their occurrence and molecular diversity in 117 domestic and free-roaming cats from the Gili Islands, Indonesia, using full-length 16S rRNA nanopore metagenomics followed by targeted PCR, sequencing, phylogenetic analysis and multilocus sequence typing (MLST). Bartonella DNA was detected in 18/117 (15.4%) cats and haemotropic Mycoplasma DNA in 40/117 (34.2%). Sequence analysis identified Bartonella henselae as the predominant species together with Bartonella clarridgeiae. MLST of B. henselae revealed three sequence types (ST1, ST16 and ST42), with ST1, a lineage reported in both feline and human isolates, predominating. Comparison with the PubMLST database showed significant geographical differences in the distribution of ST1 and ST42, supporting regional variation in the circulation of B. henselae lineages. Haemoplasma characterization identified Candidatus Mycoplasma haemominutum, Mycoplasma haemofelis, Candidatus Mycoplasma turicensis and a Mycoplasma feliminutum-like organism, comprising ten distinct sequence variants. Haemoplasma positivity was significantly associated with age, with adults showing higher positivity than younger animals (P < 0.001), whereas Bartonella infection was not associated with age, sex or island of origin. The detection of zoonotically relevant B. henselae lineages and the genetic diversity of feline haemoplasmas provide evidence of the circulation of vector-borne bacteria among cats in this tropical island ecosystem. These findings provide the first molecular epidemiological baseline for this region and contribute to understanding the circulation and genetic diversity of feline vector-borne pathogens in Southeast Asia.

Animals↗

Molecular characterization of the sucB gene encoding the immunogenic dihydrolipoamide succinyltransferase protein of Bartonella vinsonii subsp. berkhoffii and Bartonella quintana.

Members of the genus Bartonella have historically been connected with human disease, such as cat scratch disease, trench fever, and Carrion's disease, and recently have been recognized as emerging pathogens causing other clinical manifestations in humans. However, because little is known about the antigens that elicit antibody production in response to Bartonella infections, this project was undertaken to identify and molecularly characterize these immunogens. Immunologic screening of a Bartonella vinsonii subsp. berkhoffii genomic expression library with anti-Bartonella antibodies led to the identification of the sucB gene, which encodes the enzyme dihydrolipoamide succinyltransferase. Antiserum from a mouse experimentally infected with live Bartonella was reactive against recombinant SucB, indicating the mounting of an anti-SucB response following infection. Antigenic cross-reactivity was observed with antiserum against other Bartonella spp. Antibodies against Coxiella burnetti, Francisella tularensis, and Rickettsia typhi also reacted with our recombinant Bartonella SucB. Potential SucB antigenic cross-reactivity presents a challenge to the development of serodiagnostic tests for other intracellular pathogens that cause diseases such as Q fever, rickettsioses, brucelloses, tularemia, and other bartonelloses.

Acyltransferases↗

Disruption of a host-parasite system following the introduction of an exotic host species.

The potential of biological invasions to threaten native ecosystems is well recognized. Here we describe how an introduced species impacts on native host-parasite dynamics by acting as an alternative host. By sampling sites across an invasion front in Ireland, we quantified the influence of the introduced bank vole (Clethrionomys glareolus) on the epidemiology of infections caused by flea-transmitted haemoparasites of the genus Bartonella in native wood mice (Apodemus sylvaticus). Bartonella infections were detected on either side of the front but occurred exclusively in wood mice, despite being highly prevalent in both rodent species elsewhere in Europe. Bank vole introduction has, however, affected the wood mouse-Bartonella interaction, with the infection prevalence of both Bartonella birtlesii and Bartonella taylorii declining significantly with increasing bank vole density. Whilst flea prevalence in wood mice increases with wood mouse density in areas without bank voles, no such relationship is detected in invaded areas. The results are consistent with the dilution effect hypothesis. This predicts that for vector-transmitted parasites, the presence of less competent host species may reduce infection prevalence in the principal host. In addition we found a negative relationship between B. birtlesii and B. taylorii prevalences, indicating that these two microparasites may compete within hosts.

Animals↗

Evaluation of the relationship between causative organisms and clinical characteristics of infective endocarditis in dogs: 71 cases (1992-2005).

OBJECTIVE: To evaluate microbiologic findings in dogs with infective endocarditis (IE) and determine whether there were differences in clinical features of disease caused by different groups of infective agents. DESIGN: Retrospective case series. ANIMALS: 71 dogs with suspected or definite IE. PROCEDURES: Medical records were reviewed for results of bacterial culture and susceptibility testing, serologic assays for vector-borne disease, and PCR testing on vegetative growths. Cases were grouped by causative organism and relationships among infectious agent group, and various hematologic, biochemical, and clinical variables were determined. Survival analyses were used to determine associations between infecting organisms and outcome. RESULTS: Causative bacteria were identified in 41 of 71 (58%) dogs. Gram-positive cocci were the causative agents in most (21/41; 51%) infections, with Streptococcus canis associated with 24% of infections. Gram-negative organisms were detected in 9 of the 41 (22%) dogs. Infection with Bartonella spp was detected in 6 of 31 (19%) dogs with negative results for microbial growth on blood culture. Aortic valve involvement and congestive heart failure were more frequent in dogs with endocarditis from Bartonella spp infection, and those dogs were more likely to be afebrile. Infection with Bartonella spp was negatively correlated with survival. Mitral valve involvement and polyarthritis were more frequent in dogs with streptococcal endocarditis. CONCLUSIONS AND CLINICAL RELEVANCE: Streptococci were the most common cause of IE and were more likely to infect the mitral valve and be associated with polyarthritis. Dogs with IE secondary to Bartonella spp infection were often afebrile, more likely to develop congestive heart failure, rarely had mitral valve involvement, and had shorter survival times.

Animals↗

Prevalence of Bartonella species and 16s rRNA gene types of Bartonella henselae from domestic cats in Thailand.

Prevalence of Bartonella infection among 275 cats in 9 sites from 4 geographical regions (northern area: Chiang Mai; central area: Kanchanaburi, Ratchaburi, and Bangkok; northeastern area: Khon Kaen, Roi Et, Ubon Ratcharthani, and Nakhonratchasima; southern area: Songkhla) of Thailand was investigated. Overall, Bartonella species were isolated from 27.6% (76 of 275) of the cats. The isolation rate varied from 12.8% (5 of 39) in Songkhla (southern area) to 50.0% (26 of 52) in Khon Kaen (northeastern area). Bartonella henselae and B. clarridgeiae were isolated from 82.9% (63 of 76) and 11.8% (9 of 76) of the Bartonella-positive cats, respectively. Coinfection with both species was found in 5.3% (4 of 76) of the bacteremic cats. Of the 67 bacteremic cats from which B. henselae was isolated, 48 (71.6%) and 13 (19.4%) were infected with only Type I and Type II, respectively. Coinfection with both types was observed in 9.0% (6 of 67) of the B. henselae-positive cats. To our knowledge, this is the first report on the presence of Bartonella infection in domestic cats from Thailand, which constitute a large reservoir of Bartonella infection in this country.

Animals↗

[Study on Bartonella vinsonii berkhoffii isolated from blood of native dogs in China].

OBJECTIVE: To isolate and identify Bartonella strains from native dogs in Shandong province in China. METHODS: EDTA-anticoagulated blood samples were collected from 71 native dogs in Yanggu county of Shandong province in March 2005. All isolates were grown on brain heart infusion agar plates containing 5% defibrinated rabbit blood. The agar plates were incubated at 37 degrees C in a humidified with 5% CO2 environment for 4 weeks or longer. All Bartonella-like isolates were examined by routine Gram and Giménez staining and then followed by polymerase chain reaction (PCR) and PCR-RFLP analysis for identification and differentiation of the isolates. Sequencing 16S rRNA, citrate synthase (gltA) gene and 16S-23S rRNA ITS were carried out and sequential similarities were calculated using the DNASTAR5 software package. The phylogenetic tree was inferred from each bootstrap sample, using the neighbor-joining methods as executed in the MEGA 3.1 software. The translation from DNA to protein were determined by DNASIS 2.5. RESULTS: The two Bartonella-like organisms (strains Q52SHD and Q64SHD) were isolated from the blood of 71 dogs. Light microscopic examination of the Gram and Giménez-stained micro-organisms showed small, short and slightly curved pleomorphic gram-negative bacilli. Amplified products of the three pairs of Bartonella genus-specific primers carried the same size as the predicted of those Bartonella species. Data from PCR-RFLP analysis showed that the two strains that having the same profiles were all different from the B. henselae type strain-16S rRNA, gltA and 16S-23S rRNA ITS sequences from the two isolates were 100.0%, 99.7% and 97.2% homologous to B. vinsonii berkhoffii. CONCLUSIONS: Based on these findings, the two isolates Q52SHD and Q64SHD were demonstrated as B. vinsonii berkhoffii. To our knowledge, this was the first report on the presence of Bartonella infection in native dogs from China, which constituted a large reservoir of Bartonella species in this country.

Animals↗

[Capreolus capreolus and Ixodes ricinus as a reservoir of Bartonella in north-western Poland].

Capreolus capreolus and Ixodes ricinus as a reservoir of Bartonella in north-western Poland. The purpose of the study was to assess the prevalence Bartonella in Capreolus capreolus from north-western Poland forest. Supplementary, ticks infesting roe deer were also screened in order to ascertain their role as vectors and reservoirs of Bartonella. The samples of blood from 98 animals from north-western Poland were PCR-screened. Bartonella DNA was detected by using primers complementary to the intergenic spacer (ITS) between the 16S and 23S rRNA genes, which is used for identification of over a dozen species of this genus. Products of three different sizes were detected: 230 bp and 290 bp may represent two strains of B. capreoli, and 190 bp may be identify as B. bovis. All three amplicons were detected in blood, the 290 bp fragment from B. capreoli was present only in ticks, Ixodes ricinus. Generally, Bartonella infection in C. capreolus amounted to 21.4% of individuals, but was much higher during the autumn-winter seasons (62%), than in spring (4.3%). The results show that C. capreolus may be a reservoir for at least two species, i.e. B. capreoli and B. bovis, and probably do not cause persistent infection in roe deer. The high percentage of infested individuals during spring (84%) and infection detected in I. ricinus (5.2%) show that ticks are reservoir and vector of Bartonella.

Animals↗

[Cat-scratch disease and other infections caused by Bartonella species].

Bartonella henselae, the causative agent of cat-scratch disease, was identified recently by DNA amplification techniques. Several other Bartonellae (most of which were called Rochalimaea before) cause disease in humans: B. bacilliformis (Carrion's disease), B. elizabethae (endocarditis) and B. quintana (bacillary angiomatosis and peliosis, chronic bacteraemia and endocarditis, trench fever). B. henselae is transmitted to humans by scratch or bite of a bacteraemic, but asymptomatic, cat, which event may be followed by regional lymphadenitis (classical cat-scratch disease), bacillary angiomatosis or peliosis of liver and spleen (in immune compromised, e.g. HIV-infected individuals) or chronic bacteraemia and endocarditis (in elderly individuals). The incidence in the Netherlands of cat-scratch disease is > 2/100,000/year. If a Bartonella infection is suspected, specific immuno-assays and polymerase chain reaction assay may be applied for diagnosis. Culture of the organism is difficult. Macrolides and tetracyclines have been shown to be effective in treatment of disseminated infections. The natural (self-limiting) course of regional lymphadenitis however is not affected by antibiotic treatment.

Aged↗

Immunopathology of Bartonella vinsonii (berkhoffii) in experimentally infected dogs.

Following natural infection with Bartonella, dogs and humans develop comparable disease manifestations including endocarditis, peliosis hepatis, and granulomatous disease. As the immunologic response to infection in these hosts has not been clearly established, data presented here was derived from the experimental infection of six specific pathogen free (SPF) beagles with a known pathogenic strain of Bartonella. Six dogs were inoculated intravenously with 10(9)cfu of B. vinsonii ssp. berkhoffii and six control dogs were injected intravenously with an equivalent volume of sterile saline. Despite production of substantial levels of specific antibody, blood culture and molecular analyses indicated that Bartonella established chronic infection in these dogs. Flow cytometric analysis of monocytes indicated impaired bacterial phagocytosis during chronic Bartonella infection. There was also a sustained decrease in the percentage of CD8+ lymphocytes in the peripheral blood. Moreover, modulation of adhesion molecule expression (downregulation of L-selectin, VLA-4, and LFA-1) on CD8+ lymphocytes suggested quantitative and qualitative impairment of this cell subset in Bartonella-infected dogs. When compared with control dogs, flow cytometric analysis of lymph node (LN) cells from B. vinsonii infected dogs revealed an expanded population of CD4+ T cells with an apparent naïve phenotype (CD45RA+/CD62L+/CD49D(dim)). However, fewer B cells from infected dogs expressed cell-surface MHC II, implicating impaired antigen presentation to helper T cells within LN. Taken together, results from this study indicate that B. vinsonii establishes chronic infection in dogs which may result in immune suppression characterized by defects in monocytic phagocytosis, an impaired subset of CD8+ T lymphocytes, and impaired antigen presentation within LN.

Animals↗