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We detected Rickettsia felis DNA in Ctenocephalides felis and Bartonella quintana DNA in 3 Pulex irritans fleas taken from a pet Cercopithecus cephus monkey in Gabon, sub-Saharan Africa. This is the first report of B. quintana in the human flea.
We identified a Bartonella quintana strain by polymerase chain reaction amplification, cloning, and sequencing of DNA extracted from lysed erythrocytes and cultured colonies grown from peripheral blood collected from a captive-bred cynomolgus monkey (Macaca fascicularis). This report describes naturally acquired B. quintana infection in a nonhuman primate.
Bartonella quintana, a pathogen that is restricted to human hosts and louse vectors, was first characterized as the agent of trench fever. The disease was described in 1915 on the basis of natural and experimental infections in soldiers. It is now recognized as a reemerging pathogen among homeless populations in cities in the United States and Europe and is responsible for a wide spectrum of conditions, including chronic bacteremia, endocarditis, and bacillary angiomatosis. Diagnosis is based on serologic analysis, culture, and molecular biology. Recent characterization of its genome allowed the development of modern diagnosis and typing methods. Guidelines for the treatment of B. quintana infections are presented.
We report a case of cat scratch disease caused by Bartonella henselae in Korea. A 25-yr-old woman developed left cervical lymphadenopathy with history of contact with a dog. The cervical lymphadenopathy persisted for 1 month and resolved gradually and spontaneously. Serologic test was not done during the acute stage of the disease. Immunofluorescent antibody test performed during the convalescent stage was positive for B. henselae. To confirm B. henselae infection, polymerase chain reaction (PCR) analysis using aspirates of cervical lymph node was performed and the presence of B. henselae DNA was demonstrated. This is the first reported case of cat scratch disease in Korea confirmed by PCR for B. henselae DNA.
The prevalences of antibodies against Bartonella henselae and Ehrlichia canis were determined in sera from 228 dogs in 5 communal lands of Zimbabwe, areas where traditional subsistence agro-pastoralism is practised. The sera were collected from apparently healthy dogs during routine rabies vaccination programmes and tested with indirect fluorescent antibody assays using B. henselae (Houston-I) and E. canis (Oklahoma) as antigens. We found reactive antibodies (> or =1:80) against B. henselae in 14% of the dogs tested. Seropositive animals were found in Bikita (41%; 17/42), Omay (13%; 6/48), Chinamora (5%; 2/38) and Matusadona (15%; 7/48). No seropositive dogs were found in Chiredzi (0%; 0/52). Antibodies reactive with E. canis (> or =1:80) were found in 34% of the dogs tested, from Bikita (88%; 37/42), Chiredzi (31%; 16/52), Omay (17%; 8/48), Chinamora (26%; 10/38) and Matusadona (15%; 7/48). Our survey shows dogs in the communal lands of Zimbabwe are frequently exposed to E. canis and B. henselae or closely related species. Further studies are indicated to determine the pathogenicity of the organisms infecting these dogs and their clinical significance.
OBJECTIVE: To determine the usefulness of an indirect immunoflourescence antibody test for antibodies to Bartonella henselae in diagnosing cat scratch disease (CSD). DESIGN AND SETTING: Retrospective case survey of 354 patients whose sera were tested for antibodies to B. henselae at Royal Perth Hospital, Perth, and the Institute of Clinical Pathology and Medical Research, Sydney. In 1994; and measurement of the background prevalence of antibodies to B. henselae. MAIN OUTCOME MEASURES: Prevalence of antibodies to B. henselae, odds of a positive titre (> or = 64) in patients with and without specific risk factors for CSD and clinical features of the disease; prevalence of antibodies to B. henselae in randomly selected blood donors. RESULTS: Demographic, clinical and cat contact data were available for 303 patients. Sixty-four (21.1%) had a positive titre, as did 53 of 98 (54%) patients with a history of cat contact and lymphadenopathy. This proportion increased to 62% (38 of 61 patients) in patients with a history of cat scratch or bite and to 90.3% (28 of 31) in those with cat contact, lymphadenopathy and histological evidence of granulomatous lymphadenitis. Patients who developed lymphadenopathy after cat contact were significantly more likely to have a positive titre than those without this history (odds ratio [OR], 20.8; 95% confidence interval [95% Cl], 9.6-46; P < 0.0001). Inclusion of a history of a cat scratch or bite significantly raised the odds of being seropositive (OR, 13.7; 95% Cl, 6.8-28.1; P < 0.0001), and the presence of granulomas on lymph node biopsy further increased the odds (OR, 124.4; 95% Cl, 19.4-1073; P < 0.0001). The prevalence of antibodies to B. henselae in random blood donors in New South Wales was about 5% (five of 102 sera samples). CONCLUSIONS: The immunofluorescence antibody test for B. henselae can be expected to be positive in just over half the patients with clinically suspected CSD, and it has a positive predictive value of 83%. In a significant number of cases the diagnosis cannot be made on the basis of the results of immunofluorescence antibody testing alone and further investigations, including lymph node biopsy, may be required.
CONTEXT: Cat scratch disease (CSD) commonly occurs secondary to Bartonella henselae infection, and the diagnosis has traditionally been made by microscopic findings, the identification of organisms by cytochemistry, and clinical history. However, cytochemical analysis tends to be very difficult to interpret, and histology alone may be insufficient to establish a definitive diagnosis of CSD. OBJECTIVE: To demonstrate the presence of B henselae in tissue suspected of involvement by CSD, using a novel polymerase chain reaction (PCR) assay. DESIGN: Isolates of B henselae (American Tissue Culture Collection 49793) and Afipia felis (American Tissue Culture Collection 49714) were cultured on blood agar and buffered charcoal yeast extract agar, respectively. DNA was isolated from these organisms and from formalin-fixed, paraffin-embedded tissue sections with involvement by CSD (8 patients). Negative controls included water, human placental tissue, and lymph node specimens from 6 patients with reactive lymphoid hyperplasia and from 2 patients with granulomatous lymphadenitis. A primer complementary to B henselae citrate synthase gltA gene sequence was designed to perform a seminested PCR amplification. For restriction fragment length polymorphism analysis, PCR products were digested by TaqI restriction enzyme and analyzed by gel electrophoresis. RESULTS: Seminested PCR analysis of the cultured isolates of B henselae, but not of A felis, showed specific amplification. However, nonnested PCR did not provide consistently positive results in tissue sections with CSD. Therefore, we used a seminested PCR, which revealed positivity in all of the cases with clinicopathologic diagnoses of CSD. None of the negative controls showed positivity. Restriction enzyme provided confirmation of the specific PCR amplification of the B henselae sequence. CONCLUSIONS: Since the amplification product has a low molecular size (<200 base pairs), this assay is useful for detection of B henselae in formalin-fixed, paraffin-embedded tissues. The seminested PCR protocol described here can be used for rapid and reliable confirmation of B henselae in samples that are histologically suggestive of CSD.
BACKGROUND: Bartonella henselae infections are closely related to numerous clinical infections of growing interest in Spain. Since immunosuppressed patients are a potential risk group for infection by this bacteria, the aim of the present was to study the seroepidemiology of B. henselae infection in a risk group (patients with HIV infection) and in a control group (donors). PATIENTS AND METHODS: In October, 1997, antibodies versus B. henselae were determined at different dilutions (cut off > or = 1:64) by immunofluorescence in 52 patients with HIV infection and 85 donors. An epidemiologic study included age, sex, smoking, alcohol intake, INVDA, HIV infection, AIDS stage, cutaneous anergy, CD4 lymphocyte count, antiretroviral treatment and chemoprophylaxis versus P. carinii. RESULTS: Nine of the patients with HIV infection (17.3%) and five donors (5.88%) presented titers > or = 1:64 with no significant differences (p = 0.06) (adjusted OR: 1.7; CI 95%: 0.34-8.54). Moreover, multiple logistic regression analysis did not show any risk or protection factor associated with B. henselae infection in patients with HIV infection. CONCLUSIONS: A high level of seroprevalence of antibodies versus B. henselae was observed in patients with HIV infection. No risk or protection factors associated with B. henselae infection in patients with HIV infection were found.
Human bartonellosis in North America is mainly associated with Bartonella henselae, and the availability of laboratory diagnostic tools has significantly heightened awareness of the spectrum of human disease that is caused by this bacterium. We detail herein examples of illness in a pediatric population which serve to confirm that B. henselae-associated disease exists in British Columbia. Seroprevalence studies among asymptomatic adults and among children with symptomatic respiratory illness of other causation demonstrated that 36.8% and 18.5% of sera, respectively, had IFA-IgG titres > or = 1:256. IFA-IgG titres did not vary significantly whether B. henselae ATCC 49793 or a local wild-type B. henselae isolate were used as substrate. An assessment of IgM response was consistent with the proposal that endemic seroprevalence is a function of past rather than recent exposure. Both clinical and serological studies are concordant in providing evidence that B. henselae is endemic in British Columbia.
Serological study of 788 blood sera, taken from residents of the Moscow region was conducted using antigens of microorganisms of the genera Rickettsia and Bartonella. The first group under examination consisted of 355 patients with diagnosed diseases of nonreckettsial nature. The second group includes 433 healthy adults working at a meat processing and packing factory. The main method used for sera survey was the indirect immunofluorescence test. In the sera taken from the first group of subjects specific antibodies to R. prowazekii, R. typhi, B. quintana, B. henselae antigens were detected in 2.3%, 5.1%, 4.0% and 2.9% of serum samples respectively. In the serum samples taken from the second group the proportion of antibodies to R. prowazekii, R. typhi, B. quintana, B. henselae antigens was different: 0.5%, 3.3%, 1.7% and 4.0% respectively. In total, specific antibodies to R. typhi and B. henselae prevailed over specific antibodies to R. prowazekii and B. quintana twofold.
The seroprevalence of Bartonella henselae among veterinary professionals in Japan was investigated by means of an immunoperoxidase (IP) test that used protein A-horseradish peroxidase conjugate. Sera were obtained from 233 veterinary professionals in the Tokyo and Chiba areas. As negative control group, sera from 155 healthy individuals (all medical students) were used. As positive control group, sera from 5 patients highly suspected of cat scratch disease (CSD) by clinical symptoms were tested. Serum antibody titers of > or = 200 to B. henselae were presumed seropositive, because the titer at which about 95.5% of all healthy individuals (148 of the 155) were negative, and 2 of the 5 suspected CSD patients' serum antibody titers were > or = 200. Of the individuals in the veterinary professionals group tested, 35 of the 233 (15.0%) were seropositive for B. henselae. Females were nearly twice as likely as males to have antibodies to B. henselae in the veterinary professionals group. Our data suggest that Japanese veterinary professionals, and in particular younger females who are veterinary assistants and animal beauticians are more often infected by B. henselae.
The authors review the cases of two patients with an atypical form of cat scratch disease. The first case exhibited ileocoecal and hepatoportal lymphadenitis, and the second case the emerging symptoms of a non-specific granulomatous process in the middle third of the right forearm, which caused a differential-diagnostic problem. Establishment of a diagnosis was possible only on the basis of the disease process and the pathological, serological and histopathological pictures of the lymphatic glands after other causes of the lymphadenopathy had been excluded. Following two weeks of antibiotic treatment, the patients were free of complaints- and symptoms. Bartonella antibody tests were performed on 5 cats living in the neighbourhood of the patients with titer determinations too in 3 cases.
We present a case of a 22-month-old child with swollen upper part of the right arm and osteolytic lesion of the right humerus, which resembled a neoplastic process. Epidemiological history revealed no scratch marks on the skin or cutaneous papule or pustule. Presumptive diagnosis of hematogenous osteomyelitis was established, but treatment with fusidic acid was unsuccessful. Histological examination of the bioptic specimen of the soft tissue swelling showed a lymph node morphology, with numerous granulomas with central stellate necrosis. Indirect immunofluorescence assay for Bartonella henselae yielded positive results. New treatment included 15 days of trimetoprime and sulfamethoxazole, followed by azithromycin for 5 days. Four months later, swelling resolved and osteolytic lesion almost completely healed with formation of surrounding sclerosis. In conclusion, cat-scratch disease without positive epidemiological history and primary cutaneous papule or pustule may be a serious diagnostic problem, but can be solved by serological and histological examination.
Bartonella henselae, an emerging pathogen bacterium, is the main causative agent of the cat scratch disease. While the first clinical descriptions were associated with immunosupressed patients, it is now more frequently observed in patients with normal immune status (endocarditis and bacteremia). Cats were found to be the only known reservoir of B. henselae. In this paper, we report the results obtained in the first study made to investigate the prevalence of B. henselae bacteremia and antibodies in domestic cats in Catalonia, Spain. Serum samples from 115 cats were tested for antibodies to B. henselae by immunofluorescent antibody testing, and 29.6% had a titer >or= 1:64. Seven B. henselae strains were isolated using standard culture techniques and amplification by a polymerase chain reaction and subsequent sequencing was performed on the intergenic spacer region between the 16 and 23S ribosomal RNA genes. Of all factors concerning the studied bacteremia rate (age, sex, habitat, presence of antibodies, contact with animals, parasites), only the presence of antibodies to B. henselae was statistically significant.
Bartonellosis, a biphasic disease caused by motile intracellular bacteria, produces in its tissue phase a characteristic dermal eruption (Verruga peruana) resulting from a pronounced endothelial cell proliferation. Bacteria are found in the interstitium and within the cytoplasm of endothelial cells (Rocha-Lima inclusion). The aim of this study was to determine if Bartonella bacilliformis produce a substance(s) that might be responsible for the vascular proliferation seen in the Verruga. This was assessed in an in vitro system using human endothelial cells and measuring proliferation as well as production of tissue type plasminogen activator after exposure to the endothelial cultures to B. bacilliformis extracts. Our results indicate that B. bacilliformis possess an activity that stimulates endothelial cell proliferation up to three times that of control. The factor(s) is specific for endothelial cells, heat sensitive, larger than 12 to 14 kd, not enhanced by heparin, has no affinity for heparin, and is precipitated by 45% ammonium sulfate. In addition, the B. bacilliformis extracts stimulate production of t-PA antigen in a concentration-dependent fashion. This activity is also heat sensitive and not lost after dialysis (12 to 14 kd). B. bacilliformis extracts, however, do not increase the production of plasminogen activator inhibitor. It was also determined that B. bacilliformis extracts stimulate the formation of new blood vessels in an in vivo model for angiogenesis. These results describe a bacterial factor(s) that stimulates two important steps in the development of new blood vessels in vitro, as well as the formation of new blood vessels in vivo. Determining the mechanism of action, combined with a complete characterization of this factor(s), may help in understanding the pathogenesis not only of the Verruga and angiogenesis in general but also the recently described Cat-Scratch-associated epithelioid hemangiomas in patients with AIDS and Kaposi sarcoma.
OBJECTIVES: To elucidate kinetics of Bartonella henselae bacteremia and IgG response, evaluate antibiotic therapy, and investigate challenge exposure in cats. ANIMALS: Specific-pathogen-free cats. PROCEDURE: Cats were inoculated with B henselae or B quintana and monitored. Convalescent cats were challenge exposed with B henselae. Amoxicillin, enrofloxacin, erythromycin, and tetracycline HCl were evaluated for effect on B henselae bacteremia. RESULTS: Cats developed B henselae bacteremia within 1 week; bacteremia persisted for longer than 2 months before subsiding spontaneously. IgG antibody titer developed shortly after onset of bacteremia; antibody co-existed with bacteremia for several weeks and remained detectable after bacteremia subsided. Cats inoculated with B quintana remained abacteremic. On challenge exposure to B henselae, cats previously infected with B henselae remained abacteremic; cats previously inoculated with B quintana supported B henselae infection. Tetracycline HCl and erythromycin depressed B henselae bacteremia; however, duration of bacteremia remained similar to that in untreated cats. Obvious signs of illness were not observed. CONCLUSIONS: Long-duration, high-titer B henselae infections were highly reproducible in cats. Convalescent cats were immune to reinfection. B quintana-inoculated cats did not have evidence of infection and were susceptible to B henselae challenge exposure. Antibiotic therapy was incompletely efficacious in terminating cat bacteremia. CLINICAL RELEVANCE: A cat with an inapparent B henselae infection must provisionally be regarded as a possible reservoir for infection for a minimum of 2 to 3 months. Convalescent cats are resistant to reinfection. Usual antibiotic therapy was not completely efficacious. Measurement of IgG antibody can be used to detect past or current infection.
Serological and epidemiological studies suggest that Bartonella henselae is the etiological agent of cat scratch disease. We designed a study to detect B. henselae in archival biopsies by polymerase chain reaction amplification of the 16S rRNA gene followed by Southern blot hybridization. Forty-two histologically defined cat scratch disease biopsies and eighteen controls were selected for blinded analysis. After testing, charts were reviewed for clinical, immunological, and microbial evidence of infection. Results were correlated with duration of illness and antimicrobial therapy. B. henselae DNA was identified in 27 of 42 (64%) histologically defined patients and 23 of 34 (68%) patients defined both clinically and histologically. There were no false positives (0 of 18). A small subset (n = 14) had cat scratch disease serological tests performed. B. henselae was identified in 8 of 10 serologically positive patients. Polymerase chain reaction detected 50% of our DNA-positive cases (most of these early in the clinical course). Southern blotting of amplicons both doubled sensitivity (detecting patients > 4 weeks into illness) and confirmed B. henselae as the causative species. Our study strongly associates B. henselae with cat scratch disease, suggesting that it may be the most likely etiological agent in the majority of patients with cat scratch disease.
The in-vitro susceptibility of 10 isolates of Bartonella henselae was assessed using the Etest. The organisms, one reference human strain and nine feline isolates, were grown on chocolate agar and the Etests read at days 5, 8 and 11. Six antibiotics, erythromycin, azithromycin, doxycycline, ciprofloxacin, rifampicin and vancomycin were evaluated. The results correlated well with published results using agar dilution. The results confirmed the high in-vitro susceptibility of B. henselae to erythromycin, azithromycin, doxycycline and rifampicin and to a lesser extent ciprofloxacin. The majority of isolates were resistant to vancomycin. Although in-vitro results of B. henselae susceptibility testing may not necessarily correlate with clinical response, the Etest may be a simpler way for laboratories to monitor for the development of resistance particularly in the setting of relapsing infection.