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Dynamics of the IncW genetic backbone imply general trends in conjugative plasmid evolution.

Plasmids cannot be understood as mere tools for genetic exchange: they are themselves subject to the forces of evolution. Their genomic and phylogenetic features have been less studied in this respect. Focusing on the IncW incompatibility group, which includes the smallest known conjugative plasmids, we attempt to unveil some common trends in plasmid evolution. The functional modules of IncW genetic backbone are described, with emphasis on their architecture and relationships to other plasmid groups. Some plasmid regions exhibit strong phylogenetic mosaicism, in striking contrast to others of unusual synteny conservation. The presence of genes of unknown function that are widely distributed in plasmid genomes is also emphasized, exposing the existence of ill-defined yet conserved plasmid functions. Conjugation is an essential hallmark of IncW plasmid biology and special attention is given to the organization and evolution of its transfer modules. Genetic exchange between plasmids and their hosts is analysed by following the evolution of the type IV secretion system. Adaptation of the trw conjugative machinery to pathogenicity functions in Bartonella is discussed as an example of how plasmids can change their host modus vivendi. Starting from the phage paradigm, our analysis articulates novel concepts that apply to plasmid evolution.

Animals↗

Molecular genetic methods for the diagnosis of fastidious microorganisms.

Technological innovations in the detection and identification of microorganisms using molecular techniques such as polymerase chain reaction (PCR) have ushered in a new era with respect to diagnostic microbiology. PCR using universal or specific primers followed by identification of amplified product, mainly by sequencing, has enabled the rapid identification of cultured or uncultured bacteria. Thus, PCR may allow quick diagnosis of infections caused by fastidious pathogens for which culture could be extremely difficult. However, several pitfalls, such as false positives, have been observed with PCR, underlining the necessity to interpret the results obtained with caution. At present, certain improvements in the molecular genetic methods may be helpful for the diagnosis of infectious diseases. Indeed, the recent development of bacterial genome sequencing has provided an important source of potential targets for PCR, allowing rational choice of primers for diagnosis and genotyping. In addition, the development of new techniques such as real-time PCR offers several advantages in comparison to conventional PCR, including speed, simplicity, reproducibility, quantitative capability and low risk of contamination. Herein, we review the general principles of PCR-based diagnosis and molecular genetic methods for the diagnosis of several hard-to-culture bacteria, such as Rickettsia spp., Ehrlichia spp., Coxiella burnetii, Bartonella spp., Tropheryma whipplei and Yersinia pestis.

Actinomycetales↗

Sonicated diagnostic immunoblot for bartonellosis.

Two simple Bartonella bacilliformis immunoblot preparation methods were developed. Antigen was prepared by two different methods: sonication of whole organisms or glycine extraction. Both methods were then tested for sensitivity and specificity. Well-defined control sera were utilized in the development of these diagnostic immunoblots, and possible cross-reactions were thoroughly examined. Sera investigated for cross-reaction with these diagnostic antigens were drawn from patients with brucellosis, chlamydiosis, Q fever, and cat scratch disease, all of whom were from regions where bartonellosis is not endemic. While both immunoblots yielded reasonable sensitivity and high specificity, we recommend the use of the sonicated immunoblot, which has a higher sensitivity when used to detect acute disease and produces fewer cross-reactions. The sonicated immunoblot reported here is 94% sensitive to chronic bartonellosis and 70% sensitive to acute bartonellosis. In a healthy group, it is 100% specific. This immunoblot preparation requires a simple sonication protocol for the harvesting of B. bacilliformis antigens and is well suited for use in regions of endemicity.

Bartonella↗

What do we (not) know about the human bartonelloses?

The human bartonelloses are a group of diseases with a rapidly increasing clinical spectrum. Well known manifestations such as Carrion's disease, trench fever, cat-scratch disease, and bacillary angiomatosis are examples of Bartonella sp. infection. Along with these diseases, recurrent bacteremia, endocarditis, septicemia, erythema nodosum, erythema multiforme, trombocytopenic purpura and other syndromes have been reported having been caused by bacteria of this genus. The infectious process and the pathogenesis of these microorganisms are poorly understood. The bartonelloses may have a benign and self-limited evolution in a host, or a potentially fatal one. These bacteria can provoke a granulomatous or an angioproliferative histopathologic response. As these diseases are not yet well defined, we have reviewed the four main human bartonelloses and have examined unclear points about these emergent diseases.

Angiomatosis, Bacillary↗

Verruga peruana: an infectious endemic angiomatosis.

Microbial-related dysplastic and neoplastic angiomatous proliferative processes are seen with increased frequency, particularly in the acquired immunodeficiency syndrome (AIDS). The microbial-encoded or -induced mediators of angiopathogenesis in AIDS-associated Kaposi's sarcoma and bacillary angiomatosis are actively being sought. The present review addresses the historical, epidemiologic, clinical, etio- and histopathogenic aspects of the verruga peruana (VP). VP is a disease thus far endemic to high Andean valleys and characterized by dermal angioblastic proliferation in association with reactivation of latent Bartonella bacilliformis organisms. VP closely resembles AIDS-associated angiopathogenic manifestations at the clinical, histopathologic, and etiologic levels and therefore has been proposed as a model for the study of angiogenesis and endothelial cell dysplasia and neoplasia. Moreover, the recent epidemic outbreaks in endemic areas, the increased frequency of international travel to the region, the variable incubation period, and the possibility of not recognizing VP due to its rarity further underscore the relevance of studying this rare disorder and of including it in the differential diagnosis of angiomatous-proliferative disorders.

Acquired Immunodeficiency Syndrome↗

Assessing the association between the geographic distribution of deer ticks and seropositivity rates to various tick-transmitted disease organisms in dogs.

OBJECTIVE: To determine whether the geographic distribution of deer ticks (Ixodes scapularis) was associated with the distribution of dogs seropositive for various tick-transmitted disease organisms (ie, Borrelia burgdorferi, Rickettsia rickettsii, the human granulocytic ehrlichiosis [HGE] agent, Ehrlichia canis, and Bartonella vinsonii subsp berkhoffii). DESIGN: Serologic survey. SAMPLE POPULATION: Serum samples from 277 dogs in animal shelters and veterinary hospitals in Rhode Island. RESULTS: Overall, 143 (52%) dogs were seropositive for B burgdorferi, 59 (21.3%) were seropositive for R rickettsii, 40 (14.4%) were seropositive for the HGE agent, 8 (2.9%) were seropositive for E canis, and 6 (2.2%) were seropositive for B vinsonii. Regression analysis indicated that the natural logarithm of nymphal deer tick abundance was correlated with rate of seropositivity to the HGE agent and to B burgdorferi but not to rate of seropositivity to R rickettsii, E canis, or B vinsonii. Percentages of samples seropositive for B burgdorferi, R rickettsii, the HGE agent, and E canis were significantly higher for samples from the southwestern part of the state where ticks in general and deer ticks in particular are abundant than for samples from the northern and eastern portions of the state, where ticks are relatively rare. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that all 5 disease agents are in Rhode Island and pose a risk to dogs and humans. Knowledge concerning tick distributions may be useful in predicting the pattern of disease associated with particular tick species and may aid diagnostic, prevention, and control efforts.

Animals↗

Unraveling mysteries associated with cat-scratch disease, bacillary angiomatosis, and related syndromes.

The search for the infectious agents responsible for cat-scratch disease, bacillary angiomatosis, and related syndromes has a long and often circuitous history. Recognition of the etiologic agents and a new understanding of the fundamental features of the epidemiology and natural history of modern day Bartonella (formerly Rochalimaea)-associated diseases culminate a multipartite story that combines clinical medicine, traditional microbiology, and novel technological approaches to solve a long-standing enigma.

AIDS-Related Opportunistic Infections↗

An epidemic of Oroya fever in the Peruvian Andes.

Between February and October 1987, a febrile illness killed 14 persons and seriously affected at least 14 others in Shumpillan, a remote Peruvian mountain village of 353 people. The illness was characterized by fever, headache, chills, and pallor. The fatality rate of untreated cases was 88%. The patients, 71% of whom were male, were 1-75 years of age. Fatal illnesses progressed from lethargy to coma to death in 3-60 days. Patients treated empirically with chloramphenicol survived. Bartonella bacilliformis was isolated from the whole blood of 3 patients. A serologic study revealed a high prevalence of antibodies to B. bacilliformis in the villagers. It is concluded that the villagers suffered from an epidemic of Oroya fever.

Adolescent↗

Bartonellosis in Ecuador: serosurvey and current status of cutaneous verrucous disease.

Human bartonellosis is a classically biphasic disease caused by infection with the alpha-2 Proteobacteria Bartonella bacilliformis, which is phylogenetically related to the etiologic agents of cat scratch disease, bacillary angiomatosis, and trench fever. In Ecuador, typical bartonellosis has remained endemic for the past century in highland provinces near the Peruvian border. During the past six years, public health officials have noted an increasing number of atypical cases in which monophasic verrucous cutaneous disease is the only clinical manifestation. Epidemiologic, immunologic, histopathologic, and molecular biological studies have confirmed the presence of sporadic, atypical bartonellosis in residents of the lowland province of Manabi, where archeologic evidence exists of bartonellosis in pre-Colombian times. Between 1987 and 1995, 11 cases of cutaneous bartonellosis were investigated and serologic studies were done on 224 persons from five villages, two lowland and three highland. In the lowland village of Pajan in the province of Manabi, there was a 21% seropositivity proportion in contacts of index cases. These combined data suggest that bartonellosis is significantly under-reported due to the existence of mild clinical disease, possibly associated with less virulent bacterial strains, which are now disseminating or re-emerging in previously disease-free areas.

Antibodies, Bacterial↗

An outbreak of acute bartonellosis (Oroya fever) in the Urubamba region of Peru, 1998.

During May 1998, we conducted a case-control study of 357 participants from 60 households during an outbreak of acute bartonellosis in the Urubamba Valley, Peru, a region not previously considered endemic for this disease. Blood and insect specimens were collected and environmental assessments were done. Case-patients (n = 22) were defined by fever, anemia, and intra-erythrocytic coccobacilli seen in thin smears. Most case-patients were children (median age = 6.5 years). Case-patients more frequently reported sand fly bites than individuals of neighboring households (odds ratio [OR] = 5.8, 95% confidence interval [CI] = 1.2-39.2), or members from randomly selected households > or = 5 km away (OR = 8.5, 95% CI = 1.7-57.9). Bartonella bacilliformis isolated from blood was confirmed by nucleotide sequencing (citrate synthase [g/tA], 338 basepairs). Using bacterial isolation (n = 141) as the standard, sensitivity, specificity, and positive predictive value of thin smears were 36%, 96%, and 44%, respectively. Patients with clinical syndromes compatible with bartonellosis should be treated with appropriate antibiotics regardless of thin-smear results.

Adolescent↗

Cat-scratch disease in a child with unique magnetic resonance imaging findings.

OBJECTIVE: To report a patient with cat-scratch disease and the associated radiological findings. PATIENT: A previously health 10-year-old boy presenting with fever and joint pain. RESULTS: The presence of multiple abnormal foci in the bone marrow were noted by magnetic resonance imaging. There was no correlation with either radionuclide or conventional radiographic imaging findings. The patient's condition was diagnosed as cat-scratch disease by the demonstration of elevated antibody titers to the causative organism, Bartonella (formerly Rochalimaea) henselae. CONCLUSION: Magnetic resonance imaging may be more sensitive to the early bone marrow changes that can occur with cat-scratch disease than either radionuclide bone scan or computed tomography.

Antibodies, Bacterial↗

Cat-scratch disease-associated arthropathy.

OBJECTIVE: To characterize the articular manifestations of cat-scratch disease (CSD) and to evaluate the long-term clinical outcome of those manifestations. METHODS: A community- and hospital-based surveillance study of CSD was conducted in Israel between 1991 and 2002. CSD was defined as present in a patient when a compatible clinical syndrome and a positive confirmatory finding of Bartonella henselae (by serology and/or polymerase chain reaction) were identified. CSD patients with arthropathy (arthritis/arthralgia) that limited or precluded usual activities of daily living constituted the study group. Patients were followed up until > or =6 weeks after resolution of symptoms, or if symptoms persisted, for >/=12 months. CSD patients without arthropathy served as controls. RESULTS: Among 841 CSD patients, 24 (2.9%) had rheumatoid factor-negative arthropathy that was often severe and disabling. Both univariate and multivariate analyses identified female sex (67% of arthropathy patients versus 40% of controls; relative risk [RR] 2.5, P = 0.047), age older than 20 years (100% of arthropathy patients versus 43% of controls; RR 4.9, P = 0.001), and erythema nodosum (21% of arthropathy patients versus 2% of controls; RR 7.9, P = 0.001) as variables significantly associated with arthropathy. Knee, wrist, ankle, and elbow joints were most frequently affected. Ten patients (42%) had severe arthropathy in the weight-bearing joints, which substantially limited their ability to walk, and 4 of these patients were hospitalized. All of the patients had regional lymphadenopathy, 37.5% had nocturnal joint pain, and 25% had morning stiffness. Nineteen patients (79.2%) recovered after a median duration of 6 weeks (range 1-24 weeks), whereas 5 patients (20.8%) developed chronic disease persisting 16-53 months (median 30 months) after the onset of arthropathy. CONCLUSION: This is the first comprehensive study of arthropathy in CSD. CSD-associated arthropathy is an uncommon syndrome affecting mostly young and middle-age women. It is often severe and disabling, and may take a chronic course.

Adolescent↗

Regional lymphadenopathy in cat-scratch disease: ultrasonographic findings.

BACKGROUND: Cat-scratch disease (CSD) is considered to be an emerging disease worldwide and is caused by Bartonella henselae, a gram-negative bacterium introduced by a scratch or bite of a cat. The most common clinical manifestation is regional lymphadenopathy, but clinical recognition may be difficult, as atypical manifestations may occur. The diagnosis is confirmed with serologic testing and histology is rarely needed. This paper is based on our experience with the use of ultrasonography in the diagnosis of CSD. OBJECTIVE: The aim of this study was to describe the sonographic and color Doppler appearances of regional lymphadenopathy in CSD, as this has not widely reported in the literature. MATERIALS AND METHODS: Forty-seven patients (average 9.4 years) were included who all had serologically and/or histologically proven CSD and had been studied using US early in the clinical course. All had a positive history of exposure to cats and exhibited regional lymphadenopathy. RESULTS: US showed large hypoechoic adenopathy with some transmission enhancement and high vascularization on color-flow Doppler imaging. In 30 patients, abdominal US was also performed and splenic and/or hepatic granulomata were found in 10. CONCLUSIONS: In our experience, sonography and especially color-Doppler and power-Doppler sonography was helpful in the diagnosis of CSD. We believe it should be used in the initial study of children with regional lymphadenopathy, and serologic testing should be performed when CSD is suspected.

Adolescent↗

Cat-scratch disease simulating a malignant process of the chest wall.

UNLABELLED: Cat-scratch disease is a well-known cause of regional adenopathy in immunocompetent children. Rarely, patients may present with symptoms simulating a neoplastic disease. The case of a 12-year-old boy with fever, swelling of the chest wall and hepatosplenic nodules is reported. Histological analysis of biopsy specimen obtained from the chest wall lesion and the liver revealed granulomatous reaction without malignant cells. Serological investigations were positive for Bartonella species. The symptoms and the lesions disappeared after oral antibiotic therapy (ciprofloxacin) was started. The patient remained symptom-free 12 months later. CONCLUSION: The present case emphasizes the wide spectrum of clinical manifestations associated with cat-scratch disease which should be investigated as differential diagnosis of manifestations suggesting a neoplastic disease in children.

Bartonella↗

An unusual outcome in a child with hepatosplenic cat-scratch disease.

Typical cat-scratch disease (Bartonella henselae infection) in an immunocompetent child is usually associated with a history of scratch, bite or intimate contact with a cat. Most patients develop a non-tender papule in the scratch line after three to ten days. This may persist for only a few days or as long as two to three weeks. During the next two weeks or more, regional lymph nodes that drain the area gradually enlarge and then slowly resolve in more than 10% of patients. The nodes develop overlying erythema and may suppurate. Atypical forms of cat-scratch disease occur in a minority of cases and are characterized by ocular or neurological manifestations, hepatosplenic involvement, vertebral osteomyelitis, endocarditis etc. Immunocompromised individuals with B. henselae infection may develop bacillary angiomatosis, bacillary peliosis, and relapsing bacteremia. There have been several reports of hepatosplenic granulomas caused by B. henselae in immunocompetent children. We report a case of a 6-year-old boy with the hepatosplenic form of cat-scratch disease. Despite early diagnosis and long-term antimicrobial treatment, splenectomy could not be avoided.

Anti-Bacterial Agents↗

Pattern visual evoked potentials in eyes with disc swelling due to cat scratch disease-associated neuroretinitis.

PURPOSE: To evaluate optic nerve function by pattern visual evoked potentials (VEPs) in eyes with optic disc swelling due to neuroretinitis associated with cat scratch disease (CSD). METHODS: Four eyes of four patients with marked optic disc swelling resembling optic neuritis but diagnosed serologically as CSD received systemic steroid treatment. VEPs elicited by black and white checkerboard stimuli created on a TV monitor were recorded before the treatment. RESULTS: The visual acuity (VA) in the affected eyes was decreased to 20/50 in two eyes and finger counting in two eyes at their initial visits. Ophthalmoscopic examination revealed neuroretinitis characterized by severe optic disc swelling, chorioretinal exudates, and macular edema in all eyes. Anti-Bartonella henselae serum antibody was markedly elevated in all patients confirming the diagnosis of CSD. The P100 of the transient VEPs was only mildly reduced without a delay in the implicit times in three eyes and only slightly delayed in the other eye. The steady-state VEPs were mildly reduced in two eyes and phase-reversed in other two eyes. The VA fully recovered after systemic steroid treatment in all patients. CONCLUSIONS: Although all examined patients showed marked swelling of the optic disc and visual decrease, the pattern VEPs were not affected as severely as in idiopathic optic neuritis. However, the degree of change of the pattern VEPs varied among patients.

Adult↗

Visceral manifestation of cat scratch disease in children. A consequence of altered immunological state?

A 12-year-old girl with a 2-month history of fever and abdominal pain was admitted to our hospital. Ultrasound and CT scans of the abdomen showed multiple hypoechoic lesions of liver and spleen. Screening for zoonosis revealed high positive titers to Bartonella henselae. T-cell deficiency was demonstrated and remained almost unchanged during a follow-up of 11 months. A review of the literature shows that disseminated visceral affection is a rare presentation of cat scratch disease (CSD) in childhood and adolescence. Further immunological investigations are needed in more patients with CSD to confirm whether an altered immunological state may be responsible for the atypical visceral manifestation of CSD.

Animals↗