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PubMed2003

Identification and characterization of the DdlB, FtsQ and FtsA genes upstream of FtsZ in Bartonella bacilliformis and Bartonella henselae.

Homologues of the cell division protein FtsZ were previously identified in Bartonella bacilliformis and Bartonella henselae. We report herein that ftsZ is located at the distal end of an operon that includes ddlB, ftsQ, and ftsA. These genes code for homologues of D-alanine D-alanine ligase, an enzyme involved in cell wall biosynthesis, and FtsQ, and FtsA, which are involved in cell division. The DdlB, FtsQ, and FtsA proteins from Bartonella species are most homologous to proteins in closely related species from the Order Rhizobiales, such as Brucella sp., Agrobacterium tumefaciens, and M. loti. The organization of the genes within the ddlB-ftsZ operon of B. bacilliformis and B. henselae (5'ddlB-ftsQ-ftsA-ftsZ 3') is similar to that of Mesorhizobium loti and Escherichia coli. We report the localization of three promoter regions within the ddlB-ftsA sequence of B. bacilliformis that may enhance the transcription of ftsZ mRNA. A promoter region was also identified upstream of the ddlB gene.

Amino Acid Sequence↗
PubMed1996

Granulomatous hepatitis and necrotizing splenitis due to Bartonella henselae in a patient with cancer: case report and review of hepatosplenic manifestations of bartonella infection.

Bacillary angiomatosis and the related disorders of bacillary peliosis hepatis and bacillary splenitis are manifestations of infection with Bartonella henselae and Bartonella quintana in immunocompromised persons. B. henselae infection, but not B. quintana infection, is linked to contact with cats and is presumed to cause visceral cat-scratch disease. We reports a case of visceral infection by B. henselae in an adult patient with cancer who was receiving chemotherapy and had had no contact with a cat or dog. The patient--whose illness was eventually diagnosed on the basis of findings of histologic, polymerase chain reaction, and serological studies--was treated with doxycycline and rifampin, and the infection resolved. In addition, 41 cases of documented or suspected bartonella infection of the liver, spleen, or both in immunocompetent or immunocompromised hosts are reviewed.

Antineoplastic Agents↗
PubMed2002

Comparison of in-house and commercial slides for detection by immunofluorescence of immunoglobulins G and M against Bartonella henselae and Bartonella quintana.

We compared the sensitivities and specificities of indirect fluorescent antibody tests developed in our laboratory and commercially available from Focus Technologies (FT; formerly MRL Diagnostic) for detection of serum antibodies to Bartonella spp. Serum samples tested were from patients with culture- or PCR-confirmed Bartonella quintana or B. henselae infections causing cat scratch disease (CSD), chronic bacteremia, or endocarditis. At a cutoff titer of 64, the FT test had higher sensitivity than our in-house test in detecting anti-B. henselae immunoglobulin G (IgG) antibodies in CSD patients (91.2 versus 52.9%; P < 0.001). The specificity in serum samples from 85 control patients was, however, lower with the FT test (87%) than with the in-house test (98.8%) (P = 0.002). A cutoff titer of 128 improves the specificity for the FT test but lowers the sensitivity to 85%. For patients infected with B. henselae, our in-house test, but not the FT test, enabled endocarditis to be detected more reliably. With the in-house test, titers of IgG against B. henselae of >/=1,024 were found only in endocarditis patients and not in CSD patients. With the FT test, 19.1% of CSD patients had titers of IgG against B. henselae of >/=1,024 (P < 0.001). Our in-house technique also improved detection of anti-B. quintana antibodies in homeless patients with endocarditis. IgG titers of >/=1,024 were present in 75% of serum samples, but only in 16.7% of serum samples with the FT test (P = 0.004). Since each test has advantages over the other, the serological diagnosis of Bartonella infections would benefit if both tests were used concurrently.

Antibodies, Bacterial↗
PubMed1999

Molecular analysis of riboflavin synthesis genes in Bartonella henselae and use of the ribC gene for differentiation of Bartonella species by PCR.

The biosynthesis pathway for riboflavin (vitamin B(2)), the precursor of the essential cofactors flavin mononucleotide and flavin adenine dinucleotide, is present in bacteria and plants but is absent in vertebrates. Due to their conservation in bacterial species and their absence in humans, the riboflavin synthesis genes should be well suited either for detection of bacterial DNA in human specimens or for the differentiation of pathogenic bacteria by molecular techniques. A DNA fragment carrying the genes ribD, ribC, and ribE, which encode homologues of riboflavin deaminase (RibD) and subunits of riboflavin synthetase (RibC and RibE), respectively, was isolated from a plasmid-based DNA library of the human pathogen Bartonella henselae by complementation of a ribC mutation in Escherichia coli. Sequence analysis of the ribC gene region in strains of B. henselae, which were previously shown to be genetically different, revealed that the ribC gene is highly conserved at the species level. PCR amplification with primers derived from the ribC locus of B. henselae was used to isolate the corresponding DNA regions in B. bacilliformis, B. clarridgeiae, and B. quintana. Sequence analysis indicated that the riboflavin synthesis genes are conserved and show the same operon-like genetic organization in all four Bartonella species. Primer oligonucleotides designed on the basis of localized differences within the ribC DNA region were successfully used to develop species-specific PCR assays for the differentiation of B. henselae, B. clarridgeiae, B. quintana, and B. bacilliformis. The results obtained indicate that the riboflavin synthesis genes are excellent targets for PCR-directed differentiation of these emerging pathogens. The PCR assays developed should increase our diagnostic potential to differentiate Bartonella species, especially B. henselae and the newly recognized species B. clarridgeiae.

Amino Acid Sequence↗
PubMed2002

Experimental infection of domestic cats with Bartonella koehlerae and comparison of protein and DNA profiles with those of other Bartonella species infecting felines.

Bartonella koehlerae, a recently described feline Bartonella species, was isolated from two naturally infected cats in northern California. We experimentally infected domestic cats with B. koehlerae to establish the microbiological and immunological characteristics of this infection in cats and to compare it to infections with those caused by B. henselae and B. clarridgeiae. Four cats were inoculated intradermally with B. koehlerae (8.6 x 10(7) to 3.84 x 10(8) CFU/ml). None of the cats presented any obvious clinical signs, but all cats developed bacteremia, which peaked at 3.36 x 10(4) to 1.44 x 10(6) CFU/ml of blood between day 14 and day 36 postinoculation. B. koehlerae-inoculated cats had a bacteremia duration (mean, 74 days) shorter than did cats inoculated with B. clarridgeiae (mean, 324 days) (P = 0.03). None of the four cats inoculated with B. koehlerae had bacteremia relapse. As shown by enzyme-linked immunosorbent assay (ELISA) using B. koehlerae outer membrane protein (OMP) antigens, the four cats developed a species-specific antibody response, and ELISA testing using other feline Bartonella OMP antigens showed statistically lower optical density values. All four cats developed similar antibody reactivity patterns to B. koehlerae OMP antigens as seen by Western blotting, each with at least 20 seroreactive protein bands. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, protein profile differences were observed for both whole-cell lysate and OMPs from B. koehlerae, compared with B. henselae and B. clarridgeiae. B. koehlerae was more closely related to B. henselae than to B. clarridgeiae by protein profile, and this relatedness was also confirmed by analysis of the genomic DNA profiles by pulsed-field gel electrophoresis.

Animals↗
PubMed2004

[Prevalence of Bartonella henselae in stray and domestic cats in different Italian areas: evaluation of the potential risk of transmission of Bartonella to humans].

Bartonella henselae is the major etiological agent of Cat Scratch Disease in humans. Cats act as the natural reservoir of B. henselae and can transmit the infection to humans by bite or scratch. The diffusion of B. henselae was evaluated by seroprevalence and bacteremic status in different stray cat populations located in nine areas of Northern Italy. A total of 1585 cats were tested by blood culture and 361 (23%) resulted bacteremic; 1416 out off 1585 cats were also tested for Bartonella henselae antibodies and 553 (39%) resulted seropositive. The molecular typing of the isolates showed that 26% of bacteremic cats were infected with B. henselae type I, 52% with B. henselae type II, 16% were co-infected with both and 5% infected with B. Clarridgeiae. Moreover 165 domestic cats were tested by blood culture and serological test (IFA test cut-off: 1:64). 35 cats (21%) resulted bacteremic and 49 (43.5%) were seropositive. The molecular typing of the Bartonella isolates of the domestic cats showed that 45% of bacteremic cats were infected with B. henselae type I, 36.5% with B. henselae type II, 12% were coinfected with both and 6% infected with B. Clarridgeiae. For a completely evaluation of health status of the cat for B. henselae infection, the authors suggest both blood culture and serological tests. Nevertheless a nonbacteremic cat with positive serology result should be reevaluated for possible recurrent bacteremia.

Animals↗
PubMed2004

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↗
PubMed1997

Detection of Bartonella henselae and Bartonella quintana by a simple and rapid procedure using broad-range PCR amplification and direct single-strand sequencing of part of the 16S rRNA gene.

OBJECTIVE: To detect directly Bartonella henselae and Bartonella quintana using culture-independent, molecular techniques, and to evaluate a simple and rapid procedure that allows uncultivable bacteria to be detected in usually sterile clinical specimens in a diagnostic laboratory. METHODS: From four clinical specimens proven to contain B. henselae (n=3) or B. quintana (n=1) DNA, part of the 16S rRNA gene was amplified using the polymerase chain reaction (PCR) and broad-range bacterial primers followed by reamplification and direct, single-strand sequencing. The partial 16S rRNA sequences were compared to reference sequences in databases. RESULTS: Similarities between sequences derived from clinical samples and those of B. henselae and B. quintana, respectively, were in the range 98.7--100%, indicating a strong association to the genus Bartonella. Intraspecies sequence variations within the B. henselae sequences were observed. CONCLUSIONS: The method described is a rapid, sensitive and reliable tool to generate partial 16S rRNA sequences of B. henselae and B. quintana directly from normally sterile clinical specimens. It is compatible with adequate prevention of contamination as is needed for diagnostic purposes, and it possesses the potential to detect other pathogens, including those as yet unknown.

Journal Article↗
PubMed1997

Comparison of the abilities of proteins from Bartonella bacilliformis and Bartonella henselae to deform red cell membranes and to bind to red cell ghost proteins.

Infections in humans by Bartonella bacilliformis, but not Bartonella henselae, are characterized by invasion of red cells. Supernatants of culture medium from B. bacilliformis and B. henselae each contain a protein which causes invagination of membranes of human red cells and formation of intracellular vacuoles. These two proteins are very similar in molecular mass, heat stability and mechanism of action. B. henselae does not bind to human red cells, but human red cell ghost membrane proteins were recognized by both bacteria, five by B. bacilliformis and the same five, and one additional protein by B. henselae. Two of these proteins had molecular masses consistent with actin and spectrin. Actin binds to five electroblotted outer membrane proteins from B. henselae and four of these proteins are retained on an actin-Sepharose column.

Bacterial Adhesion↗
PubMed1995

Bartonella henselae and Bartonella quintana adherence to and entry into cultured human epithelial cells.

Bartonella henselae expresses pili phenotypically similar to type 4 pili. B. henselae pilus expression undergoes phase variation with multiple passages. Low-passage-number, piliated B. henselae adhered to and invaded HEp-2 cells to a greater extent than did multiply passaged B. henselae with reduced pilus expression. Pili may be a pathogenic determinant for Bartonella species.

Bacterial Adhesion↗
PubMed2003

Occurrence of Bartonella henselae and Bartonella quintana in a healthy Greek population.

The purpose of this study was to determine the prevalence of IgM and IgG antibodies against Bartonella henselae and B. quintana in a healthy Greek population using a commercially available immunofluorescent test (Focus test). Five hundred healthy individuals were divided by sex into four age groups and three groups according to contact with cats. IgM antibodies were not detected in any of the subjects examined, while 99 (19.8%) and 75 (15%) were IgG seropositive to B. henselae and to B. quintana, respectively. No statistical difference in the seropositivity was observed among these groups. The IgG antibody titers ranged from 1/64 to 1/256 for B. henselae and from 1/64 to 1/512 for B. quintana. A high percentage (12.4%) of cross-reactivity between the two species was observed. Our data show that the prevalence of both Bartonella species in Greece is high. However, low IgG antibody levels are not sufficient evidence of active infection.

Adolescent↗
PubMed2004

Prevalence of Bartonella clarridgeiae and Bartonella henselae in domestic cats from France and detection of the organisms in erythrocytes by immunofluorescence.

The prevalence of Bartonella infection in a pet cat population from France was found to be 8.1% (8 of 99 cats). The intraerythrocytic location of Bartonella clarridgeiae is shown for the first time, and we show that immunofluorescence detection of the organism in erythrocytes correlates with the number of bacteria in blood.

Animals↗
PubMed2004

Prevalence of Bartonella henselae in Italian stray cats: evaluation of serology to assess the risk of transmission of Bartonella to humans.

Bartonella henselae is the major etiological agent of cat scratch disease in humans. Cats act as the natural reservoir of B. henselae and can transmit the infection to humans by a bite or scratch. The prevalence of B. henselae in cat populations was evaluated by serological and bacteriological tests. A total of 769 stray cats from three urban and three rural areas in northern Italy were sampled between January 1999 and December 2000. The positive and the negative predictive values of serological tests with respect to bacteremic status were evaluated. Tests of a total of 140 cats (18%) resulted in detection of bacteremia. A total of 540 cats were tested by serology; 207 (38%) were seropositive. Of the 531 cats tested by both methods, the results for 65 (12.2%) showed both bacteremia detection and seropositivity. The molecular typing of the isolates showed that 20.6% of bacteremic cats were infected with B. henselae type I strain, 61.1% were infected with B. henselae type II, and 18.3% were coinfected with both. A statistically significant difference in antibody and bacteremia prevalences among geographical areas was detected. Statistical analysis showed no association between characteristics such as seroprevalence-bacteremic status, sex, general health status, and the presence of ectoparasites. The negative predictive value of serological test was 84.7%, and the positive predictive value was 31.8%. Receiving operator characteristic analysis of the data showed that serological tests had a low predictive value in relation to the bacteremic status of a cat; in surveys aimed at assessing the real risk of B. henselae infection in a human population, therefore, we suggest the use of blood culture as the reference test. Nevertheless, both blood culture assays and serological tests for Bartonella infection should be performed for a complete evaluation of the health status of cats.

Animals↗
PubMed2001

Evaluation and use of a nested polymerase chain reaction assay in cats experimentally infected with Bartonella henselae genotype I and Bartonella henselae genotype II.

Cats have been shown to be infected with Bartonella henselae genotype I, B. henselae genotype II, and B. clarridgeiae. Feline bartonellosis infections and the strains involved in these infections are important in both veterinary and human medicine. Nucleic acid amplification methods such as polymerase chain reaction (PCR) are being used in both research and diagnostics as tools for understanding many infectious diseases. Bartonella bacteremia in cats is detected by blood culture; however, because of the limitations of culture (delayed turnaround time and sensitivity limits), PCR may be a more efficient method for identifying infected cats. Three distinct PCR assays that could differentiate among B. henselae genotype I, B. henselae genotype II. and B. clarridgeiae were developed and used to detect as few as 3.2 organisms. Fourteen cats experimentally infected with B. henselae genotype I and B. henselae genotype II were followed by bacterial culture and PCR through the course of infection, including periods of primary and relapsing bacteremia. The PCR assay was positive in 11 of the 14 cats for periods of 1-9 weeks after culture became negative. Of the 223 blood specimens that were culture negative, the PCR assay was positive in 38 (17%) of the specimens. Two of the 14 cats developed relapsing bacteremia. The 2 B. henselae genotypes were amplified in the cats and the bacteremic phase of these infections as determined by PCR lasted for a longer period than previously determined by culture. Using laboratory assays such as PCR to understand the strains involved in feline bartonellosis and the course of the infection is important in the understanding of these zoonotic agents.

Animals↗
PubMed1996

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↗
PubMed2004

[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↗
PubMed2006

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↗