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PubMed1995

Strategy to detect and identify Bartonella species in routine clinical laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat.

We wished to develop a cost-effective, rapid strategy to detect and identify Bartonella species in the clinical laboratory and to determine the prevalence of Bartonella infection in the Houston veteran population. Bartonella colonies were identified by colony morphology, Gram stain, RapID ANA, repetitive extragenic palindromic-PCR (REP-PCR) and whole-cell fatty acid (CFA) analysis, and these methods were compared for their usefulness. A new test order for "Rochalimaea culture" (the genus Bartonella was previously known as the genus Rochalimaea) was instituted, and in addition, all blood specimens submitted for fungal culture (obtained in an isolator tube) were processed for Bartonella culture. Over a 16-month period we isolated Bartonella henselae from only 0.4% (2 of 533) of total cultures but from 1% (2 of 204) of human immunodeficiency virus-positive patients. After sufficient growth, identification of the Bartonella isolates to the species level could be obtained in 2 days. The REP-PCR allowed discrimination of all known species, whereas CFA analysis distinguished all except B. henselae and Bartonella quintana. The RapID ANA results failed to differentiate between B. henselae and B. quintana, and results for other species differed by only one or two tests. Blood obtained from a kitten which had been introduced into the household of one patient 2 months before the onset of fever yielded a Bartonella strain which was shown to be different from the strain from the patient and distinct from other Bartonella species by a combination of REP-PCR, CFA, and growth characteristics. Subsequent analysis of the citrate synthase gene sequence showed only an 86% similarity with any of the other known Bartonella species, suggesting that this isolate represents a distinct, previously uncharacterized species of Bartonella.

AIDS-Related Opportunistic Infections↗
PubMed1998

Comparison of different DNA fingerprinting techniques for molecular typing of Bartonella henselae isolates.

Seventeen isolates of Bartonella henselae from the region of Freiburg, Germany, obtained from blood cultures of domestic cats, were examined for their genetic heterogeneity. On the basis of different DNA fingerprinting methods, including pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic consensus (ERIC)-PCR, repetitive extragenic palindromic (REP) PCR, and arbitrarily primed (AP)-PCR, three different variants were identified among the isolates (variants I to III). Variant I included 6 strains, variant II included 10 strains, and variant III included only one strain. By all methods used, the isolates could be clearly distinguished from the type strain, Houston-1, which was designated variant IV. A previously published type-specific amplification of 16S rDNA differentiated two types of the B. henselae isolates (16S rRNA types 1 and 2). The majority of the isolates (16 of 17), including all variants I and II, were 16S rRNA type 2. Only one isolate (variant III) and the Houston-1 strain (variant IV) comprised the 16S rRNA type 1. Comparison of the 16S rDNA sequences from one representative strain from each of the three variants (I to III) confirmed the results obtained by 16S rRNA type-specific PCR. The sequences from variant I and variant II were identical, whereas the sequence of variant III differed in three positions. All methods applied in this study allowed subtyping of the isolates. PFGE and ERIC-PCR provided the highest discriminatory potential for subtyping B. henselae strains, whereas AP-PCR with the M13 primer showed a very clear differentiation between the four variants. Our results suggest that the genetic heterogeneity of B. henselae strains is high. The methods applied were found useful for typing B. henselae isolates, providing tools for epidemiological and clinical follow-up studies.

AIDS-Related Opportunistic Infections↗
PubMed1999

Semiquantitative species-specific detection of Bartonella henselae and Bartonella quintana by PCR-enzyme immunoassay.

Bartonella henselae is the main causative agent of cat-scratch disease, and both B. henselae and Bartonella quintana cause angioproliferative disorders such as bacillary angiomatosis. To increase the sensitivity of Bartonella detection by PCR and to improve the species differentiation, we developed a semiquantitative, species-specific PCR-based enzyme immunoassay (EIA). The 16S rRNA gene was selected as the target sequence. Internal nucleotide sequences derived from the amplified 16S rRNA region were used to develop species-specific oligonucleotide probes for B. henselae and B. quintana. Biotin-labeled PCR products were immobilized on streptavidin-coated microtiter plates, hybridized to a digoxigenin-labeled probe, and detected with antidigoxigenin peroxidase conjugate. No cross-hybridization with other Bartonella or non-Bartonella species was observed. This EIA was as sensitive as dot blot hybridization and was 10 times more sensitive than visualization of PCR products on agarose gels. Serial dilutions of B. henselae and B. quintana suspensions demonstrated that an optical density (OD) of approximately 0.200 was equivalent to 5 CFU in the reaction mixture. By comparing the OD of the bacterial dilutions with that obtained from clinical specimens we could determine that the number of CFU in clinical samples ranged from 10(3) to 10(6) CFU/ml. The PCR-EIA developed in the present study is a rapid, sensitive, and simple method for the diagnosis of B. henselae and B. quintana infections.

Bartonella henselae↗
PubMed2001

Aortic valve endocarditis in a dog due to Bartonella clarridgeiae.

We report the first documented case of endocarditis associated with Bartonella clarridgeiae in any species. B. clarridgeiae was identified as a possible etiological agent of human cat scratch disease. Infective vegetative valvular aortic endocarditis was diagnosed in a 2.5-year-old male neutered boxer. Historically, the dog had been diagnosed with a systolic murmur at 16 months of age and underwent balloon valvuloplasty for severe valvular aortic stenosis. Six months later, the dog was brought to a veterinary hospital with an acute third-degree atrioventricular block and was diagnosed with infective endocarditis. The dog died of cardiopulmonary arrest prior to pacemaker implantation. Necropsy confirmed severe aortic vegetative endocarditis. Blood culture grew a fastidious, gram-negative organism 8 days after being plated. Phenotypic and genotypic characterization of the isolate, including partial sequencing of the citrate synthase (gltA) and 16S rRNA genes indicated that this organism was B. clarridgeiae. DNA extraction from the deformed aortic valve and the healthy pulmonic valve revealed the presence of B. clarridgeiae DNA only from the diseased valve. No Borrelia burgdorferi or Ehrlichia sp. DNA could be identified. Using indirect immunofluorescence tests, the dog was seropositive for B. clarridgeiae and had antibodies against Ehrlichia phagocytophila but not against Ehrlichia canis, Ehrlichia ewingii, B. burgdorferi, or Coxiella burnetii.

Animals↗
PubMed2004

Molecular and cellular basis of bartonella pathogenesis.

The genus Bartonella comprises several important human pathogens that cause a wide range of clinical manifestations: cat-scratch disease, trench fever, Carrion's disease, bacteremia with fever, bacillary angiomatosis and peliosis, endocarditis, and neuroretinitis. Common features of bartonellae include transmission by blood-sucking arthropods and the specific interaction with endothelial cells and erythrocytes of their mammalian hosts. For each Bartonella species, the invasion and persistent intracellular colonization of erythrocytes are limited to a specific human or animal reservoir host. In contrast, endothelial cells are target host cells in probably all mammals, including humans. Bartonellae subvert multiple cellular functions of human endothelial cells, resulting in cell invasion, proinflammatory activation, suppression of apoptosis, and stimulation of proliferation, which may cumulate in vasoproliferative tumor growth. This review summarizes our understanding of Bartonella-host cell interactions and the molecular mechanisms of bacterial virulence and persistence. In addition, current controversies and unanswered questions in this area are highlighted.

Animals↗
PubMed1991

Abscesses of the neck in infants and young children. A review of 112 cases.

A review of 112 patients 5 years of age and younger with cervical abscesses is reported. Staphylococcus aureus and group A beta-hemolytic Streptococcus were cultured most often: in 39% and 17% of patients, respectively. Sixteen patients (14%) had unusual abscesses such as infected congenital cysts, cat-scratch disease, or myocobacterial abscesses. One patient had a necrotizing infection with group A streptococci and anaerobic streptococci. Intravenous antibiotic therapy was used in 104 patients, with 94% of these patients receiving a penicillin derivative, usually an antistaphylococcal penicillin. Ninety-six percent of the patients had incision and drainage of their abscesses; 8 patients required more than one incision and drainage. Two patients required airway intervention, 1 by intubation and 1 by trachetomy. Most pediatric cervical abscesses respond well to appropriate intravenous antibiotic therapy and incision and drainage.

Abscess↗
PubMed1989

Pathologic correlation of the unknown solid parotid mass in children.

A parotid mass in a pediatric patient is stated to have an approximately 50% likelihood of being malignant. To evaluate our experience in light of this suggested incidence, we conducted a retrospective study, which included 22 patients treated over an 8-year period, 1979 to 1987. Patients' ages ranged from 16 months to 19 years, and all patients presented with an unknown, solid parotid mass. One patient had a malignancy, mucoepidermoid carcinoma. Eight patients were diagnosed with benign pleomorphic adenoma. Thirteen patients had an inflammatory process consisting of one of the following conditions: cat-scratch disease (4); atypical mycobacteria (4); benign follicular hyperplasia of a lymph node (4); and toxoplasmosis (1). The histologic findings in this series suggest that unknown solid parotid masses that occur in children or adolescents are frequently the result of inflammatory conditions and are not likely to be malignant. We therefore recommend a reconsideration of the reputed estimation of malignancy in the parotid gland in children. The majority of pathologic conditions discovered, however, are still best diagnosed and treated by surgical excision.

Adenoma↗
PubMed2000

Seroprevalence of Bartonella henselae and Toxoplasma gondii among healthy individuals in Thailand.

The seroprevalence of Bartonella henselae and Toxoplasma gondii among apparently healthy individuals, mainly blood donors, in Thailand was investigated by an indirect fluorescent antibody technique and by a latex agglutination test, respectively. Of 163 serum samples examined, 9 (5.5%) were found to be positive for B. henselae-IgG, 2 (1.2%) for B. henselae-IgM, and 5 (3.1%) for the T. gondii antibody. No significant difference was observed between male and female samples in the serological test with either B. henselae or T. gondii. The age of individuals with B. henselae-IgG was distributed from the 20s to the 70s, and B. henselae-IgM was found in the individuals of the 30s and 60s. The age of T. gondii positive samples ranged from the 20s to the 60s. In this study, the prevalence of B. henselae infection among healthy individuals in Thailand was serologically demonstrated for the first time.

Adolescent↗
PubMed2006

Bartonella spp. in deer keds, Lipoptena mazamae (Diptera: Hippoboscidae), from Georgia and South Carolina, USA.

Deer keds, Lipoptena mazamae (Diptera: Hippoboscidae), were collected from white-tailed deer (Odocoileus virginianus) and humans in Georgia and South Carolina, USA (1 October 2001-6 January 2005) and screened for the presence of DNA from Bartonella spp. Forty deer keds were screened for Bartonella spp. by polymerase chain reaction using primers specific to the riboflavin synthase gene (ribC) of Bartonella. Bartonella species closely related to Bartonella schoenbuchensis and to the etiologic agent of cat-scratch disease (Bartonella henselae) were detected in 10 keds and one ked, respectively.

Animals↗
PubMed2001

[New and emerging rickettsial and bartonella infections].

There have been recently reports on over 10 new and resurgent rickettsioses and bartonelloses in different countries, which reflects both socioeconomic processes in society and a higher methodological level of indication and identification of causative agents. In 1991, the author' laboratory, N. F. Gamaleya Research Institute of Experimental Medicine, Russian Academy of Medical Sciences, established the etiology of the new rickettsiosis Astrakhan spotted fever. It separated and studied 2 strains of Rickettsia sp. nov. from patients and 8 ones from the carrier the Ixodes tick Rhipicephalus pumilio. It is suggested that the natural focus has transformed to the anthropurgic one due to technogenic environmental pollution. The annual increase in morbidity rates (2000 cases in 1983 to 2000) and its area are a challenge to public health care and medical science. The paper presents data on the new bartonellosis cat-scratch disease (caused by Bartonella henselae) detected not only in Russia. There is also information on tick-borne rickettsiosis, epidemic typhus, and trench fever as resurgent infections.

Bartonella↗