Characterization of the first Bartonella henselae strain isolated from a cat in Italy

Bartonella henselae has been identified and characterized for the first time in Italy. A strain, designed Pavia-1, was isolated from the blood of a cat whose owner developed cat scratch disease (CSD). Pavia-1 and two American B. henselae strains (Houston-1, ATCC 49882, type I and strain 269608, UC Davis, type II) were compared by whole-cell fatty analysis (CFA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for protein profiles, Western immunoblotting (WB) for reactivity with polyclonal antibodies, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), type-specific 16S rRNA PCRs, and pulsed-field gel electrophoresis (PFGE). Bartonella clarridgeiae (ATCC 51734) was also included for comparison. Pavia-1 was identified as a B. henselae type I. PFGE allowed differentiation between B. clarridgeiae and B. henselae and furthermore, between all the B. henselae strains. The fingerprints of PFGE observed for Pavia-1 were distinct from those of B. henselae type II and also of Houston-1, suggesting that the two type I strains derived from two different clones. These results show the capability of B. henselae to develop genotypic variability between genetically related strains.     

Peliosis hepatis in a dog infected with Bartonella henselae

A 6-year-old spayed female Golden Retriever was examined because of generalized weakness and abdominal distention. Abdominal ultrasonography revealed a large quantity of peritoneal fluid. In addition, the liver appeared larger than normal and contained multiple, small, nodular masses and cyst-like structures. Abdominal exploratory surgery was performed, and 5 L of serosanguineous peritoneal fluid was removed. Gross lesions were not found in the stomach, kidneys, intestines, adrenal glands, or urinary bladder. There were diffuse cystic nodules in all liver lobes. The dog did not recover from anesthesia. A diagnosis of peliosis hepatis was made on the basis of gross and histologic appearance of the liver. A polymerase chain reaction assay revealed Bartonella henselae DNA in liver specimens. To our knowledge, this is the first report of molecular evidence of B henselae infection in a dog with peliosis hepatis.     

Gammopathy in a Spanish dog infected with Bartonella henselae

Generalised pyogranulomatous disease and hyperviscosity syndrome associated with a presumed monoclonal gammopathy was diagnosed in a three-year-old intact female Pomeranian. The Bartonella henselae antibody titer was 1:64 and Bartonella species DNA was amplified from the splenic tissue. Monoclonal gammopathies in dogs are typically associated with plasma cell and lymphoid dyscrasias and other inflammatory or infectious diseases such as ehrlichiosis and leishmaniosis. Based on this case report, infection with Bartonella species should also be added to the differential diagnoses for gammopathy in dogs. To the authors' knowledge, this is the first report of molecular evidence of Bartonella species infection in a sick dog in Spain.     

Detection of Bartonella henselae and Bartonella clarridgeiae DNA in hepatic specimens from two dogs with hepatic disease

A 4-year-old Basset Hound and a 6-year-old Doberman Pinscher were referred for diagnostic evaluation following documentation of persistently increased hepatic enzyme activities and hepatic dysfunction. Histologic evaluation of hepatic biopsy specimens from the 2 dogs revealed granulomatous hepatitis in the Basset Hound and lymphocytic hepatitis with fibrosis and copper accumulation in the Doberman Pinscher. No etiologic agents were identified histologically. Bartonella henselae DNA was subsequently amplified from hepatic tissue from the Basset Hound and Bartonella clarridgeiae was amplified from hepatic tissue from the Doberman Pinscher. Amplification was performed with a polymerase chain reaction assay incorporating primers that target a portion of the 16S-23S rRNA intergenic spacer region. Both dogs were treated with azithromycin, in combination with a variety of other medications and herbal treatments, and improved clinically. Identification of Bartonella DNA in these dogs indicates the need for future prospective studies to determine the clinical relevance of Bartonella spp infection in dogs with hepatic disease.     

Bartonella henselae infection in a dog with recalcitrant ineffective erythropoiesis

Ineffective erythropoiesis was diagnosed in an 8‐year‐old male castrated Labrador Retriever. Despite treatment with immunosuppressive therapy for suspected immune‐mediated erythrocyte maturation arrest, resolution of the nonregenerative anemia was not achieved. Following documentation of Bartonella henselae bacteremia by Bartonella alpha proteobacteria growth medium (BAPGM) enrichment blood culture, immunosuppressive therapy was discontinued, and the anemia resolved following prolonged antibiotic therapy. Bartonella immunofluorescent antibody testing was negative, whereas B henselae western blot was consistently positive. The contribution of B henselae bacteremia to ineffective erythropoiesis remains unknown; however, the potential role of B henselae in the pathophysiology of bone marrow dyscrasias warrants additional investigation.     

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.     

Prevalence of Bartonella henselae, Bartonella clarridgeiae and the 16S rRNA gene types of Bartonella henselae among pet cats in Japan

The authors investigated bacteriologically the prevalence of Bartonella infection among 690 pet cats derived from 10 private animal hospitals in six cities (Sapporo, Hokkaido Prefecture; Sendai, Miyagi Prefecture; Joetsu, Niigata Prefecture; Fujisawa, Kanagawa Prefecutre; Kyoto, Kyoto Prefecture; Sanda, Hyogo Prefecutre) and 4 counties (Mishima, Osaka Prefecture; Hikawa, Shimane Prefecture; Aira, Kagoshima Prefecture; Shimajiri, Okinawa Prefecture) located from the north to the south of Japan. Bartonella species were isolated from 7.2% (50/690) of all the cats examined. No Bartonella species were isolated from the cats in Sapporo or Sendai. The isolation rate varied from 2% in Joetsu and Sanda to 20% in Shimajiri. Bartonella clarridgeiae was isolated from two of 50 cats in Kyoto, three of 50 in Mishima and one of 50 in Shimajiri, but not in cats from the other cities or counties. Though the cats of Joetsu, Fujisawa, Kyoto, Sanda, Aira and Shimajiri were infected with either B. henselae or B. clarridgeiae, one of eight infected cats in Mishima was harboring both Bartonella species. Type I of 16S rRNA gene was the predominant type among the isolates of B. henselae, but only one isolate derived from Shimajiri was found to be of type II. Prevalence of B. clarridgeiae and the 16S rRNA gene type of B. henselae among cats in Japan was demonstrated for the first time in this investigation.     

Prevalence of Bartonella henselae and Bartonella clarridgeiae in cats and dogs in Korea

Blood, saliva, and nail samples were collected from 54 dogs and 151 cats and analyzed for the presence of Bartonella henselae with a novel nested polymerase chain reaction (PCR) method. Bartonella (B.) henselae was detected in feral cat blood (41.8%), saliva (44.1%), and nail (42.7%) samples. B. henselae was also detected in pet cat blood (33.3%), saliva (43.5%), and nail (29.5%) samples and in pet dog blood (16.6%), saliva (18.5%), and nail (29.6%) samples. Nine samples were infected with B. clarridgeiae and 2 were co-infected with B. henselae and B. clarridgeiae of blood samples of dogs. This report is the first to investigate the prevalence of B. henselae and B. clarridgeiae in dogs and cats in Korea, and suggests that dogs and cats may serve as potential Bartonella reservoirs.     

Detection of Bartonella henselae DNA in two dogs with pyogranulomatous lymphadenitis

CASE DESCRIPTION: 1 dog evaluated because of inappetence and lameness of the left hind limb of 1 day's duration and 1 dog evaluated because of inappetence, fever, and lymphadenopathy of 2 weeks' duration. CLINICAL FINDINGS: Histologic examination of excisional biopsy specimens from lymph nodes revealed pyogranulomatous lymphadenitis in both dogs. Quantitative real-time PCR assays detected Bartonella henselae DNA in blood samples and affected lymph node specimens from both dogs. Antibodies against B. henselae were not detected via immunofluorescent antibody testing during active disease in either dog. TREATMENT AND OUTCOME: 1 dog recovered after 6 weeks of treatment with doxycycline (5 mg/kg [2.3 mg/lb], p.o., q 12 h), whereas the other dog recovered after receiving a combination of azithromycin (14.5 mg/kg [6.6 mg/lb], p.o., q 24 h for 21 days), doxycycline (17.3 mg/kg [7.9 mg/lb], p.o., q 24 h for 4 weeks), and immunosuppressive corticosteroid (prednisone [3 mg/kg {1.4 mg/lb}, p.o., q 24 h], tapered by decreasing the daily dose by 25% every 2 weeks) treatment. CLINICAL RELEVANCE: B. henselae is implicated as a possible cause or a cofactor in the development of pyogranulomatous lymphadenitis in dogs. In dogs with pyogranulomatous lymphadenitis, immunofluorescent assays may not detect antibodies against B. henselae. Molecular testing, including PCR assay of affected tissues, may provide an alternative diagnostic method for detection of B. henselae DNA in pyogranulomatous lymph nodes.     

Bartonella henselae bacteraemia in domestic cats from Auckland

Bartonella henselae causes most cases of cat scratch disease, a self-limited localised lymphadenopathy illness of humans. Bartonella henselae also causes disseminated cutaneous and visceral disease in immunocompromised people. Cat blood (1-5 ml) collected from cats in the Auckland area was processed and plated on to 5% sheep blood brain heart infusion agar and incubated at 35 degrees C in 5% CO2 for 14 days. Bartonella henselae was identified by colony morphology, Gram's stain, twitching motility, biochemical tests and molecular methods. Eight of 48 cats (17%) had Bartonella bacteraemia. Species-specific probes and biochemical profiles identified all isolates as B. henselae. Infected cats pose a risk to humans they lick, scratch or bite. People should be made aware of the risk cats pose.     

Bilateral mandibular pyogranulomatous lymphadenitis and pulmonary nodules in a dog with Bartonella henselae bacteremia

This report describes a 2-year-old collie dog with pulmonary nodules, visualized by computed tomographic (CT) scan, with evidence of Bartonella henselae bacteremia and pyogranulomatous lymphadenitis. Clinical signs resolved with antimicrobial therapy.     

Investigation of Bartonella henselae in cats in Ankara, Turkey

The purpose of this study was to determine Bartonella henselae prevalance in cats in Ankara. Whole bloods and sera collected from 256 cats were investigated for the presence feline Bartonella species by culture and sera were tested for the presence of antibodies against B. henselae IgG using immunofluorescence assay. Bartonella species were isolated by blood culture from 24 (9.4%) cats. Bartonella isolates were subjected to restriction fragment length polymorphism (RFLP) by using TaqI and HhaI endonucleases to identify species. Twenty-one isolates were determined as B. henselae and three of 24 isolates were determined as Bartonella clarridgeiae with RFLP. The bacteraemia prevalence and seroprevalence of B. henselae IgG antibodies in cats was detected as 8.2% and 18.6% respectively. This is the first report on B. henselea and B. clarridgeiae in cats in Turkey.     

Experimental infection of domestic cats with passaged genotype I Bartonella henselae

The influence of in vitro passage on Bartonella henselae pathogenesis in cats has not been thoroughly evaluated. Our objective was to examine the bacterial kinetics and humoral immune responses in cats experimentally infected with three different in vitro passages of B. henselae F1, a genotype I strain of feline origin. The F1 strain was in vitro passaged 20 and 40 times, and each was inoculated into a group of 5 cats. The kinetics of bacteremia and the feline humoral immune response to bacterial antigens were compared to a previous study involving a group of six cats inoculated with the original F1 strain. Among the three groups of cats, the kinetics of bacteremia profiles and the humoral immune responses to B. henselae lysates were similar. The influence of passage on bacterial membrane proteins was examined. In vitro passage altered the expression of 4/17 (23.5%) bacterial membrane proteins and 6/15 (40%) bacterial membrane antigens. An association between poor seroreactivity to three lysate antigens (15-, 18- and 45kDa), prolonged bacteremia and decreased serum bactericidal activity was noted. Our data show that in vitro passage of B. henselae did not alter the kinetics of bacteremia, including the occurrence of relapsing bacteremia, in experimentally infected cats. This suggests that highly passaged strains may not be suitable for future vaccination studies. Furthermore, in vitro passage results in phenotypic and antigenic changes in the bacterial membrane protein profile, which warrants caution in the interpretation of studies involving passaged B. henselae strains.     

Clinicopathologic findings in dogs seroreactive to Bartonella henselae antigens

OBJECTIVE: To assess the potential clinical relevance of seroreactivity to Bartonella henselae antigens in dogs. ANIMALS: 40 dogs seroreactive to B henselae and 45 dogs that did not seroreact to B henselae. PROCEDURE: A case-control study was conducted. Clinical and clinicopathologic findings were extracted from medical records of each dog. RESULTS: Statistical differences were not detected between dogs seroreactive or nonseroreactive to B henselae when analyzed on the basis of disease category or results of hematologic, biochemical, urine, or cytologic analysis. However, seroreactivity to B henselae antigens was detected in 2 of 4 dogs with a clinical diagnosis of granulomatous meningoencephalitis, 3 of 4 dogs with immune-mediated hemolytic anemia, 3 of 4 dogs with infective endocarditis, 2 of 3 dogs with lymphoid neoplasia, and 5 of 10 dogs with polyarthritis. Additionally, seroreactivity to B henselae antigens was detected in 18 of 34 thrombocytopenic dogs and 14 of 27 dogs with neutrophilia. CONCLUSIONS AND CLINICAL RELEVANCE: Significant associations were not detected between seroreactivity to B henselae and various diseases. Prospective epidemiologic studies investigating specific diseases, such as meningoencephalitis or polyarthritis, and specific hematologic abnormalities, such as immunemediated hemolytic anemia or thrombocytopenia, should be conducted to further define the potential clinical relevance of antibodies against B henselae in dogs. IMPACT FOR HUMAN MEDICINE: Bartonella organisms are increasingly reported as pathogens that induce are increasingly reported as pathogens that induce chronic infections in humans and dogs. Dogs may serve as natural candidates for future study of the disease in humans.