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PubMed2005

Evasion of Toll-like receptor 5 by flagellated bacteria.

Toll-like receptor 5 (TLR5) recognizes an evolutionarily conserved site on bacterial flagellin that is required for flagellar filament assembly and motility. The alpha and epsilon Proteobacteria, including the important human pathogens Campylobacter jejuni, Helicobacter pylori, and Bartonella bacilliformis, require flagellar motility to efficiently infect mammalian hosts. In this study, we demonstrate that these bacteria make flagellin molecules that are not recognized by TLR5. We map the site responsible for TLR5 evasion to amino acids 89-96 of the N-terminal D1 domain, which is centrally positioned within the previously defined TLR5 recognition site. Salmonella flagellin is strongly recognized by TLR5, but mutating residues 89-96 to the corresponding H. pylori flaA sequence abolishes TLR5 recognition and also destroys bacterial motility. To preserve bacterial motility, alpha and epsilon Proteobacteria possess compensatory amino acid changes in other regions of the flagellin molecule, and we engineer a mutant form of Salmonella flagellin that evades TLR5 but retains motility. These results suggest that TLR5 evasion is critical for the survival of this subset of bacteria at mucosal sites in animals and raise the intriguing possibility that flagellin receptors provided the selective force to drive the evolution of these unique subclasses of bacterial flagellins.

Amino Acid Sequence↗
PubMed1998

Culture-negative endocarditis caused by Bartonella henselae.

A 4-year-old girl presented with clinical evidence of infective endocarditis involving her aortic valve, but blood cultures were sterile. Serologic studies and analysis of resected valve by immunohistochemistry and polymerase chain reaction established the diagnosis of Bartonella henselae endocarditis. Clinicians should be aware that B. henselae can cause apparent culture-negative endocarditis in children.

Antibodies, Bacterial↗
PubMed2000

[Neck abscess as primary manifestation of head and neck carcinoma: implications for diagnostic management].

BACKGROUND: Neck abscesses and deep neck infections are common diseases in the field of otorhinolaryngology. However, malignant lymph node metastases presenting as abscesses are uncommon and have rarely been described up to now. PATIENTS: A retrospective review between March 1997 and August 1999 was conducted of 40 patients with deep neck abscesses. RESULTS: In five patients (12.5%) the histological diagnostics revealed a malignancy. From these, in 3 cases the cervical abscess was the primary clinical symptom of an oropharyngeal as well as a hypopharyngeal carcinoma. With further 2 patients, the neck-abscess was located as first clinical sign of a metachronous lymph node metastases after oropharyngeal carcinoma. Aerobic bacteria were recovered in 18 patients, anaerobes alone in 3 and mixed aerobic and anaerobic bacteria in 6 patients. Bartonella henselae was recovered in 7, Mycobacterium tuberculosis in 3 and Actinomyces israelii in 1 patient. CONCLUSIONS: A biopsy of the abscess wall is recommended in establishing the diagnosis. The indication for a simultaneous panendoscopy--especially in patients with typical risk for malignancies--to exclude a primary tumor in the aerodigestive tract should be regarded generously. Neck abscesses should be considered in the differential diagnosis of head and neck carcinoma.

Abscess↗
PubMed2003

[Hepatosplenic localization of cat scratch disease in immunocompetent adults. Two cases].

The infective agent responsible for cat scratch disease, Bartonella henselae, is a rare cause of hepatic granulomatosis in immunocompetent adults. Clinical features include a prolonged fever or more typical symptoms such as lymphadenopathy associated with painful hepatomegaly and a fever following a cat scratch or bite. Images of micronodular hepatosplenic lesions on abdominal ultrasonography or computed tomography scan along with epithelioid granulomas in a liver biopsy can suggest this diagnosis. It is established with a serology by indirect immunofluorescence or by ELISA and/or the presence of Bartonella henselae DNA evidenced by PCR in the liver biopsy. We report two cases of hepatosplenic localizations of cat scratch disease in a 41-year-old woman and a 44-year-old man presenting asthenia and fever associated with a biological inflammatory syndrome and elevated liver enzymes.

Adult↗
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↗
PubMed1998

[Infective endocarditis caused by unusual microorganisms].

All series of infective endocarditis had a variable proportion of cases without an etiologic agent because all cultures were negative. New microbiologic techniques have permitted the discovery of the role of many microorganisms in infective endocarditis. C. burnetii is an increasing causative agent of subacute infective endocarditis. In the diagnosis, to the detection of antiphase-I antibodies, immunohistochemical, molecular techniques and cellular cultures have been added. Total cure is difficult to obtain. The combination of doxicicline plus ciprofloxacin for at least 3 years has been proposed as the treatment of choice. Surgery must be reserved for patients with cardiac insufficiency. Less than 2% of cases of acute brucellosis are complicate with infective endocarditis. Infective endocarditis produces serious and rapid valvular destruction with high mortality rates if valve surgery is not performed. For medical treatment at least 3 active agents are required. Bartonella has recently been described as an etiologic agent of infective endocarditis. It mainly affects to homeless people living in poor hygienic conditions. The aortic valve is most commonly involved and, frequently, valve insufficiency requires valve replacement. Blood culture isolation needs long incubation periods. Parenteral nutrition, immunosuppression, wide spectrum antibiotic regimens, intravenous drug addiction and cardiovascular surgery are risk factors previously described in the development of fungal endocarditis. C. albicans and Aspergillus spp. are most frequent etiologic agents. Infective endocarditis should be suspected in any patient with systemic fungal disease. Blood cultures are often negative except for Candida spp. Peripheral emboli and large vegetations are frequent. Mortality is high, antifungal therapy combined with surgery is the treatment of choice. Legionella, Mycoplasma, Chlamydia, Mycobacteria, viruses are potential agents of infective endocarditis, and difficult to diagnose because of special culture requirements. Epidemiological clues, serologic and molecular techniques and blood cultures could identify them.

Anti-Bacterial Agents↗
PubMed2000

Prevalence of serologic evidence of cat scratch disease in patients with neuroretinitis.

OBJECTIVE: To determine the prevalence of Bartonella henselae seropositivity in patients with a clinical diagnosis of neuroretinitis. DESIGN: Retrospective, clinic-based, cross-sectional study. PARTICIPANTS: Eighteen consecutive patients seeking treatment at the Casey Eye Institute from November 1993 through November 1998 who had neuroretinitis. METHODS: The billing and photographic records of the Casey Eye Institute were searched for patients with a primary or secondary diagnosis of neuroretinitis or Leber's idiopathic stellate neuroretinitis. Charts were then reviewed to determine the results of B. henselae antibody titers and other pertinent clinical information. MAIN OUTCOME MEASURES: Results of B. henselae serologic testing. RESULTS: Fourteen of 18 patients with neuroretinitis had serologic studies. Nine of the 14 tested patients (64.3%) were found to have elevated IgM or IgG for B. henselae, suggesting current or past infection. Patients with positive serologic analysis results tended to have worse vision at presentation. There were no other obvious differences between seropositive and seronegative groups in this study, including duration or quality of recovery. CONCLUSIONS: At our tertiary care ophthalmology institution, most tested patients with neuroretinitis had evidence of past or present cat-scratch disease based on positive serologic analysis for B. henselae, a much greater prevalence than is expected to be found in the general population or in patients with idiopathic uveitis. Further study is indicated to clarify the prevalence of cat-scratch disease in neuroretinitis and the role and efficacy of antibiotics in treatment.

Adolescent↗
PubMed1997

Epidemiologic evaluation of the risk factors associated with exposure and seroreactivity to Bartonella vinsonii in dogs.

OBJECTIVES: To determine seroprevalence to Bartonella vinsonii subsp berkhoffii in a population of sick dogs from North Carolina and Virginia and to evaluate potential risk factors associated with increased likelihood of exposure to the organism. SAMPLE POPULATION: Serum samples from 1,920 sick dogs. PROCEDURE: An indirect fluorescent antibody assay was performed on each sample, and the end-point antibody titer was recorded. A case (seropositive) was defined as a dog with reciprocal titer > or = 64, and a control (seronegative) was defined as a dog with reciprocal titer < 16 that was referred within 0 to 3 days of referral of a corresponding case. From this population, 207 dogs (69 cases and 138 controls) were included in a case-control seroepidemiologic study. RESULTS: 3.6% (69/1,920) of the dogs were seropositive to B vinsonii subsp berkhoffii. Results of the case-control study indicated that seropositive dogs were more likely to live in rural environments, frequently on a farm, were free to roam the neighborhood, and were considered to be predominantly outdoor dogs. Moreover, seropositive dogs were 14 times more likely to have a history of heavy tick exposure. After analysis of the case-control study, a more detailed examination of banked sera from dogs with known tick exposure was performed. High correlation was found between sero-reactivity to B vinsonii and seroreactivity to E canis or B canis (36.0 and 57.1%, respectively). Sera derived from dogs experimentally infected with E canis or R rickettsii did not cross react with B vinsonii antigen. CONCLUSIONS AND CLINICAL RELEVANCE: Several potential risk factors are associated with canine exposure to B vinsonii. Rhipicephalus sanguineus, the tick vector for E canis and B canis, may be involved in B vinsonii transmission among dogs.

Animals↗
PubMed2002

Identification of Bartonella bacilliformis genotypes and their relevance to epidemiological investigations of human bartonellosis.

Genotypic diversity among 26 isolates of Bartonella bacilliformis obtained from different areas of Peru, and at different times, was assessed by comparison of DNA sequences derived from 16S-23S ribosomal DNA intergenic spacer regions (ISR) and a citrate synthase gene (gltA) fragment and by amplified fragment length polymorphism (AFLP) analysis. gltA comparison divided the isolates into two groups, whereas ISR comparison revealed six sequences. AFLP analysis using a selective primer delineated five profiles that correlated well with those obtained by sequence comparison. Combination of all three data sets divided the isolates into six genotypes. One of these genotypes was common to isolates collected from a large area in western Peru that corresponded to the region of endemicity for bartonellosis; however, isolates belonging to two other genotypes were also found within this region. Two of these genotypes were found in isolates isolated more than 35 years apart. The remaining three genotypes were each specifically associated with three outbreaks of bartonellosis that have recently occurred in areas where the disease had not previously been recognized. Demonstration of the unique nature of these isolates indicates that the outbreaks with which they were associated did not result from the introduction of disease by individuals who acquired their infection in the recognized region of endemicity. The sources of these outbreaks remain unknown. A consensus approach to bacterial typing using comparative sequence analysis of multiple genetic loci and the pan-genomic sampling of AFLP appears to offer a well-supported assessment of B. bacilliformis diversity, and the genotypic differences identified appear to have epidemiological significance.

Bacterial Proteins↗
PubMed2006

Bartonella quintana coinfection with Mycobacterium avium complex and CMV in an AIDS patient: case presentation.

BACKGROUND: As a greater number of HIV-infected patients survive despite profound immunodepression due to medical progress, we face complex infection with multiple agents in AIDS-patients. CASE PRESENTATION: We report the case of an AIDS patient with a primary clinical presentation suggestive of bacillary angiomatosis. We also found in cutaneous lesions Mycobacterium avium complex and cytomegalovirus. CONCLUSION: This clinical case illustrates the possibility of multiple coinfections in AIDS patients and the need to be exhaustive in evaluating infectious diseases in severely immunocompromised patients.

AIDS-Related Opportunistic Infections↗
PubMed2005

Blood culture-negative endocarditis in a reference center: etiologic diagnosis of 348 cases.

To identify the current etiologies of blood culture-negative infective endocarditis and to describe the epidemiologic, clinical, laboratory, and echocardiographic characteristics associated with each etiology, as well as with unexplained cases, we tested samples from 348 patients suspected of having blood culture-negative infective endocarditis in our diagnostic center, the French National Reference Center for Rickettsial Diseases, between 1983 and 2001. Serology tests for Coxiella burnettii, Bartonella species, Chlamydia species, Legionella species, and Aspergillus species; blood culture on shell vial; and, when available, analysis of valve specimens through culture, microscopic examination, and direct PCR amplification were performed. Physicians were asked to complete a questionnaire, which was computerized. Only cases of definite infective endocarditis, as defined by the modified Duke criteria, were included. A total of 348 cases were recorded-to our knowledge, the largest series reported to date. Of those, 167 cases (48%) were associated with C. burnetii, 99 (28%) with Bartonella species, and 5 (1%) with rare, fastidious bacterial agents of endocarditis (Tropheryma whipplei, Abiotrophia elegans, Mycoplasma hominis, Legionella pneumophila). Among 73 cases without etiology, 58 received antibiotic drugs before the blood cultures. Six cases were right-sided endocarditis and 4 occurred in patients who had a permanent pacemaker. Finally, no explanatory factor was found for 5 remaining cases (1%), despite all investigations.Q fever endocarditis affected males in 75% of cases, between 40 and 70 years of age. Ninety-one percent of patients had a previous valvulopathy, 32% were immunocompromised, and 70% had been exposed to animals. Our study confirms the improved clinical presentation and prognosis of the disease observed during the last decades. Such an evolution could be related to earlier diagnosis due to better physician awareness and more sensitive diagnostic techniques. As for Bartonella species, B. quintana was recorded more frequently than B. henselae (53 vs 17 cases). For 18 patients with Bartonella endocarditis, the responsible species was not identified. Species determination was achieved through culture and/or PCR in 49 cases and through Western immunoblotting in 22. Comparison of B. quintana and B. henselae endocarditis revealed distinct epidemiologic patterns. The 2 cases due to T. whipplei reflect the emerging role of this agent as a cause of infective endocarditis. Because identification of the bacterium was possible only through analysis of excised valves by histologic examination, PCR, and culture on shell vial, the prevalence of the disease might be underestimated. Among patients who received antibiotic drugs before blood cultures, 4 cases (7%) were found to be associated with Streptococcus species (2 S. bovis and 2 S. mutans) through 16S rDNA gene amplification directly from the valve, which shows the usefulness of this technique in overcoming the limitations of previous antibiotic treatment. Right-sided endocarditis occurred classically in young patients (mean age, 36 yr), intravenous drug users in 50% of cases, and suffering more often from embolic complications. Finally, 5 cases without etiology or explaining factors were all immunocompetent male patients with previous aortic valvular lesions, and 3 of the 5 presented with an aortic abscess. Further investigations should be focused on this group to identify new agents of infective endocarditis.

Adolescent↗
PubMed2004

Prevalence of Borrelia burgdorferi, Bartonella spp., Babesia microti, and Anaplasma phagocytophila in Ixodes scapularis ticks collected in Northern New Jersey.

PCR analysis of Ixodes scapularis ticks collected in New Jersey identified infections with Borrelia burgdorferi (33.6%), Babesia microti (8.4%), Anaplasma phagocytophila (1.9%), and Bartonella spp. (34.5%). The I. scapularis tick is a potential pathogen vector that can cause coinfection and contribute to the variety of clinical responses noted in some tick-borne disease patients.

Anaplasma phagocytophilum↗
PubMedDate not supplied

Antinuclear antibodies can be detected in dog sera reactive to Bartonella vinsonii subsp. berkhoffii, Ehrlichia canis, or Leishmania infantum antigens.

The presence of antinuclear antibodies (ANAs) is used to support a clinical diagnosis of systemic lupus erythematosus (SLE) in dogs. However, clinicians must interpret the detection of ANAs with caution, particularly in light of increasing evidence that dogs with known bacterial and protozoal infections can have high ANA titers. Retrospectively, medical records were reviewed for all dogs that were concurrently tested for antinuclear antigens and Bartonella vinsonii (berkhoffii), Ehrlichia canis, or Rickettsia rickettsii antigens between 1990 and 2000. When analyzed on the basis of reactivity to a specific infectious agent, 75% of the B vinsonii (berkhoffii) seroreactors, 16.7% of the E canis seroreactors, and 0% of the R rickettsii seroreactors had concurrent ANAs. Subsequent prospective testing did not detect ANAs in convalescent sera from dogs experimentally infected with B vinsonii (berkhoffii), E canis, or R rickettsii. However, 10-20% B vinsonii (berkhoffii), E canis, or Leishmania infantum reactive sera from naturally infected dogs contained ANAs. In addition, 45% of sera from dogs that are reactive to multiple vectorborne organisms were more likely to contain ANAs when compared to sera from dogs reactive to only 1 test antigen. When interpreting the relevance of seroreactivity to nuclear antigens, clinicians should recognize that dogs with seroreactivity to B vinsonii (berkhoffii), E canis, or L infantum antigens (especially those with seroreactivity to more than one of these pathogens) may produce ANAs.

Animals↗
PubMed1999

Prevalence of Bartonella henselae immunoglobulin G antibodies in Singaporean cats.

BACKGROUND: Bartonella henselae causes several clinical diseases in humans. The most common infection is the classical cat-scratch disease (CSD) occurring in immunocompetent and immunocompromised children exposed to newly acquired kittens. This is the first study of B. henselae infection in Singaporean cat population. METHODS: Indirect fluorescent antibody tests were carried out on cat sera obtained from 80 cat blood samples. RESULTS: Of the cats studied in this project 47.5% tested seropositive, with high IgG titers (> or =1024) in 31 of 38 seropositive cats, 60.5% of which were males. CONCLUSIONS: Males cats had higher rates of infection than female cats. Serologic studies of cats in different parts of the United States have indicated that the prevalence of IgG antibody to B. henselae in cats is highest in regions with warm, humid climates and lowest in areas with cool, dry climates. The indirect fluorescent antibody test data for B. henselae infection in Singapore cats support this contention.

Animals↗
PubMed2004

Infective endocarditis.

Despite improvements in health care, the incidence of infective endocarditis has not decreased over the past decades. This apparent paradox is explained by a progressive evolution in risk factors; while classic predisposing conditions such as rheumatic heart disease have been all but eradicated, new risk factors for infective endocarditis have emerged. These include intravenous drug use, sclerotic valve disease in elderly patients, use of prosthetic valves, and nosocomial disease. Newly identified pathogens, which are difficult to cultivate--eg, Bartonella spp and Tropheryma whipplei--are present in selected individuals, and resistant organisms are challenging conventional antimicrobial therapy. Keeping up with these changes depends on a comprehensive approach, allying understanding of the pathogenesis of disease with the development of new drugs for infective endocarditis. Infection by staphylococci and streptococci is being dissected at the molecular level. New ideas for antimicrobial agents are being developed. These novel insights should help redefine preventive and therapeutic strategies against infective endocarditis.

Anti-Bacterial Agents↗
PubMedDate not supplied

Late onset of serologic positive titers in a patient with Parinaud's oculoglandular syndrome.

PURPOSE: To report a case of Parinaud's oculoglandular syndrome (POS) in which, despite the initially negative, a late onset of serologic positive titers was observed. METHODS: Case report. RESULTS: A 65-year-old man was examined for a foreign body in the left eye associated with granulomatous nodule on the palpebral conjuctiva, chemosis and a swollen preauricular lymph node. Despite the initially negative serologic titers to Bartonella henselae, a clinical diagnosis of POS was made and treatment was instituted initially with oral ciprofloxacin (500 mg twice a day). Three weeks later, serologic positive titers were found which confirmed the diagnosis of POS. CONCLUSION: Even though the improvements in diagnostic testing such as indirect immunofluorescence antibody, negative titles could be initially observed, confusing the diagnostic process. Other techniques (such as polymerase chain reaction (PCR) testing) should be also taken into consideration in cases with increased clinical suspicious of B. henselae indections.

Aged↗
PubMed2003

Molecular diagnosis of infective endocarditis by PCR amplification and direct sequencing of DNA from valve tissue.

We used broad-range eubacterial PCR amplification followed by direct sequencing to identify microbial pathogens in heart valve material from 29 patients with histologically confirmed infective endocarditis and 23 patients free of infective endocarditis. Microorganisms cultured by conventional techniques matched those identified by PCR in 21 cases. PCR alone identified the causative agent in three cases (Streptococcus bovis, Staphylococcus cohnii, and Coxiella burnetii), allowing better patient management. PCR corrected the initial bacteriological diagnosis in three cases (Streptococcus bovis, Streptococcus mutans, and Bartonella henselae). Among the 29 cases of histologically confirmed infective endocarditis, PCR findings were positive in 27 cases and were consistent with the bacterial morphology seen at Gram staining (26 cases) or with the results obtained by immunohistologic analysis with an anti-C. burnetii monoclonal antibody (one case). In two other cases of histologically confirmed infective endocarditis, PCR remained negative in a blood culture-negative case for which no bacteria were seen at histological analysis and in one case with visualization of cocci and blood cultures positive for Enterococcus faecalis. Ten clinical diagnoses of possible infective endocarditis were ruled out by histopathological analysis of the valves and subsequently by PCR. PCR was negative in 13 of the 14 patients in whom infective endocarditis was rejected on clinical grounds; the other patient was found to have Coxiella burnetii infective endocarditis on the basis of PCR and histopathological analysis and was subsequently included in the group of 29 definite cases. In total, PCR contributed to the diagnosis and management of infective endocarditis in 6 of 29 (20%) cases.

DNA, Bacterial↗
PubMed2002

Cutaneous bacillary angiomatosis.

Bacillary angiomatosis is characterized by unique vascular lesions caused by infection with a small Gram staining bacillus of the genus Bartonella. It usually occurs in immunocompromised persons but can also occur in immunocompetent persons. We report a case of cutaneous bacillary angiomatosis in a 5-year-old immunocompetent child. He had infected lesions on the lips, after an injury, which was followed by lesions over the knees, buttocks, near the ankles and the elbows. Diagnosis was proved on histology. The lesions cleared after administration of erythromycin for 3 months. It is well to be aware of this condition in the context of increasing prevalence of AIDS.

Angiomatosis, Bacillary↗