[Molecular Epidemiology of Bartonella henselae in Stray and Sheltered Cats of Zaragoza, Spain]. / Epidemiología molecular de Bartonella henselae en gatos callejeros y de albergue en Zaragoza, España.

OBJECTIVE: Bartonella henselae is responsible for the Cat Scratch Disease in humans, being it underdiagnosed. This study aims to detect and quantify the load of B. henselae DNA in oral and whole blood samples from stray and shelthered cats from Zaragoza (Spain), and analyze associations with epidemiological and clinical factors. METHODS: 47 cats entered in the estudy. Real time PCR was used to detect B. henselae DNA in blood and oral samples. The SPSS software was applied to the statistical analysis of positivity of paired samples and its relationship with variables as age, sex, origin, month of sampling and fleas/ticks observation in fur and clinical factors (health status and observation of oral lesions). To know the relationship between the presence in blood and oral cavity a logistic regression analysis was performed. RESULTS: A 23.40% of blood samples and the 27.65% of the oral swabs carried the B. henselae DNA. A fair agreement between paired samples was observed (kappa value = 0.33, p less than 0.05). Bacterial DNA detected in oral and blood samples were not significantly associated to any of the epidemiological and clinical factors. Positive cats having oral lesions carried higher loads (3,12/1x1,000,000 cells) of bacterial DNA in their oral cavity than those without lesions (2,58/1x1,000,000 cells) being p=0.032. CONCLUSIONS: Carriage of the B. henselae DNA in the blood samples appears not to be related with carriage of the DNA of the bacteria in mouth and vice versa. Positive cats having oral lesions carry a higher load of B. henselae DNA and may suppose a higher risk of transmission to people handling them.

OBJETIVO: Bartonella henselae produce la enfermedad del arañazo del gato en las personas y se considera infradiagnosticada. El objetivo fue detectar y cuantificar la carga de ácido desoxiribonucleico (ADN) de B. henselae en muestras de sangre y orales de gatos callejeros y de albergue de Zaragoza, España y analizar su relación con factores epidemiológicos y clínicos. METODOS: Se estudiaron 47 gatos. El ADN de B. henselae,se detectó mediante reacción en cadena de la polimerasa en tiempo real (qPCR) en sangre y muestras orales. Se usó el paquete estadístico SPSS para analizar la positividad de las muestras pareadas y su relación con factores epidemiológicos (edad, sexo, origen, mes de muestreo, presencia de pulgas/garrapatas) y clínicos (estado de salud y presencia de lesiones orales). Se realizó un análisis de regresión logística para conocer la asociación entre la presencia en sangre y cavidad oral y el resto de las variables. RESULTADOS: El 23,40% de las muestras de sangre y el 27,65% de las orales portaba el ADN de B. henselae. Se observó débil correlación de la positividad de las muestras pareadas (kappa=0,33; p inferior a 0,05). No se detectó asociación estadística entre la presencia de ADN de B. henselae en las muestras y los factores epidemiológicos y clínicos. Los gatos con lesiones orales portaban una carga más elevada de ADN (3,12/1x1.000.000 células) en la boca que los que no tenían lesiones (2,58/1por 1.000.000 células),(p=0,032). CONCLUSIONES: La detección de ADN de B. henselae en sangre no parece estar relacionada con su presencia en cavidad oral y viceversa. Los gatos positivos con lesiones orales pueden significar mayor riesgo de infección por B. henselae para las personas que los manejan.

Detection of Bartonella henselae IgM in serum of experimentally infected and naturally exposed cats.

BACKGROUND: Results of Bartonella henselae blood culture, polymerase chain reaction (PCR) assay on blood, or IgG antibody assays do not always correlate with the presence or absence of clinical disease in cats, and B. henselae IgG antibodies in serum do not always correlate with bacteremia. However, little is known concerning Bartonella spp. IgM antibodies in naturally exposed cats. HYPOTHESIS: Bartonella spp. IgM antibodies in serum are associated with fever, stomatitis, and bacteremia based on PCR assay results in experimentally infected or client-owned cats. ANIMALS: Stored sera from cats experimentally infected with B. henselae by exposure to Ctenocephalides felis, client-owned cats with and without fever, and client-owned cats with and without stomatitis were studied. METHODS: A Bartonella spp. IgM ELISA was titrated with samples from experimentally infected cats and then test sera from client-owned cats were assayed. Associations among IgM ELISA results, clinical findings, and bacteremia as defined by Bartonella spp. PCR assay were assessed. RESULTS: All experimentally infected cats developed Bartonella spp. IgM antibodies. Bartonella spp. IgM antibody assay results were not always in agreement with PCR assay results in client-owned cats (60%). Bartonella spp. DNA in blood, IgM antibodies, and IgG antibodies were not associated with the presence of fever or stomatitis. CONCLUSIONS AND CLINICAL IMPORTANCE: Because Bartonella spp. IgM antibodies as measured by this assay were not associated with fever or stomatitis and were not always in agreement with PCR assay results, there appears to be little need for assessing individual client-owned cats for this antibody class alone.

Complements do not lie.

A 74-year-old patient presented with constitutional symptoms and was found to have acute kidney injury. He was known to have a prosthetic aortic valve. He was febrile with splenomegaly and vasculitic lesions on both hands. Nephritic screen revealed strongly positive cytoplasmic-antineutrophil cytoplasmic antibodies (c-ANCA). Differential diagnosis thus included a small vessel vasculitis or infective endocarditis. Transoesophageal echocardiography demonstrated no vegetations and serial blood cultures were negative. Immunosuppression for presumed granulomatosis with polyangiitis (Wegeners granulomatosis) was therefore instituted. The patient deteriorated, requiring multi-organ support. Renal biopsy showed a proliferative glomerulopathy and complements were low. Atypical screen for culture negative endocarditis revealed a strongly positive IgG-antibody titre against Bartonella henselae. Immunosuppression was discontinued and treatment for chronic Bartonellosis commenced. The patient made a remarkable recovery. His renal function quickly returned to normal, and ANCA titres and complements normalised. He was discharged home after completing a 6 week course of antibiotic therapy.

Identification of candidate proteins for the diagnosis of Bartonella henselae infections using an immunoproteomic approach.

Bartonella henselae is an emerging gram-negative facultative intracellular pathogen transmitted via Ctenocephalides felis (cat fleas) or cat scratches. Bartonellosis is present mainly in the form of cat scratch disease (CSD), bacillary angiomatosis and infective endocarditis (IE). The methods used to diagnose B. henselae rely on culturing, immunofluorescent assays and molecular techniques. The objective of the present study was to identify candidate proteins for the serodiagnosis of bartonellosis with the differential discrimination of both clinical scenarios: CSD and IE. For this, an immunoproteomic approach combined with 2-DE, immunoblotting and matrix-assisted laser desorption/ionization time-of-flight MS has been developed. Immunoproteomic profiles of sera collected from patients with CSD and IE were compared with those of blood donors. We identified several candidate proteins as phage-encoding Pap31 protein and an outer membrane protein of BH11510 that, in our view, might be useful for the serodiagnosis of bartonellosis.

Bone marrow and skin granulomatosis in a patient with Bartonella infection.

This report describes a case of granulomatous inflammation, involving the bone marrow and skin, due to Bartonella infection in an immunocompetent patient. The clinical presentation included prolonged fever, pancytopenia, rash and hepatitis. Bartonella infection should thus be added to the growing list of entities that produce marrow granulomas and fever.

[Seroepidemiology of Bartonella henselae infection in HIV-infected patients]. / Seroepidemiología de la infección por Bartonella henselae en pacientes infectados por el VIH.

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.

A chemically defined liquid medium that supports primary isolation of Rochalimaea (Bartonella) henselae from blood and tissue specimens.

Rochalimaea (Bartonella) henselae is a fastidious, slowly growing, gram-negative bacillus that is an etiologic agent of bacillary angiomatosis, cat scratch disease, and related syndromes. Accumulation of direct microbiologic evidence of the relationship between the organism and the syndromes compatible with cat scratch disease has been hindered by the difficulties in the primary isolation of the organism from infected tissue specimens. A chemically defined liquid medium was developed to support the growth of Rochalimaea species to facilitate study of the organism. This medium was also used successfully to isolate R. henselae from clinical specimens from infected patients and a domestic cat. Recovery of R. henselae in this was more successful than when recovery was attempted on solid agar. This cell-free, extract-free, defined medium additionally supported the growth of Rochalimaea quintana and Afipia felis.