Detecção de anticorpos anti-Rickettsia rickettsii em cães residentes em área negligenciada no município de São Paulo, SP, Brasil / [Detection of anti-Rickettsia rickettsii antibodies in dogs living in a neglected area in São Paulo, SP, Brazil]

A febre maculosa brasileira (FMB), descrita inicialmente nos Estados Unidos como febre maculosa das Montanhas Rochosas, é uma antropozoonose relatada apenas no continente americano e causada pela bactéria Rickettsia rickettsii. No Brasil a transmissão ocorre sobretudo pela picada de carrapatos do gênero Amblyomma spp. A doença foi inicialmente descrita como de transmissão em áreas rurais e silvestres, no entanto áreas periurbanas e urbanas vêm apresentando casos, principalmente relacionados com a presença de humanos residindo em pequenos fragmentos de mata ciliar. O presente estudo teve por objetivo elucidar a dispersão da FMB nas proximidades dos reservatórios Guarapiranga e Billings, na cidade de São Paulo, SP. Para tanto, a presença de anticorpos anti-R. rickettsii, Rickettsia parkeri e Rickettsia bellii foi avaliada em cães atendidos nas campanhas de esterilização cirúrgica e residentes ao redor dos reservatórios. Foram coletadas amostras de 393 cães, e as amostras de soro foram analisadas pela reação de imunofluorescência indireta (RIFI), com ponto de corte de 1:64. Os títulos para R. rickettsii variaram de 256 a 4096, com positividade de 3,3% (13/393); para R. bellii, de 128 a 1024 e 4,1% (16/393) de positivos, e um único animal (0,25%) foi soropositivo para R. parkeri, com título de 128. Os achados permitem concluir que a região de estudo apresenta condições de se tornar uma possível área com casos de FMB, pois comporta fragmentação de Mata Atlântica, condições essas ideais para a manutenção do vetor do gênero Amblyomma já descrito na região, bem como para a presença da Rickettsia rickettsii circulante entre os cães, confirmada pela existência de anticorpos. Condutas referentes à conscientização da população por meio de trabalhos educacionais devem ser implantadas para a prevenção da doença na população da área.(AU)

Brazilian Spotted Fever (BSF), initially described in the United States as Rocky Mountain Spotted Fever, is an anthropozoonosis reported only in the Americas and caused by the bacterium Rickettsia rickettsii. In Brazil, transmission occurs mainly through tick bites of the genus Amblyomma spp. The disease was initially described as transmission of rural and wild areas; however, peri-urban and urban areas have been presenting cases, mainly related to the presence of humans residing in small fragments of riparian forest. The present study aimed to elucidate the dispersal of BSF near the Guarapiranga and Billings Reservoirs, in the city of São Paulo, SP. The presence of anti-R. rickettsii, Rickettsia parkeri and Rickettsia bellii antibodies were evaluated in dogs treated in surgical sterilization campaigns and residents around the Reservoirs. Samples were collected from 393 dogs and serum samples were analyzed by indirect immunofluorescence reaction (RIFI) with a cutoff of 1:64. The titles for R. rickettsii varied from 256 to 4096 with a positivity of 3.3% (13/393); for R. bellii from 128 to 1024 and 4.1% (16/393) of positive and a single animal (0.25%) was seropositive for R. parkeri with a titre of 128. The findings allow us to conclude that the study region has conditions to become a possible area with BSF cases, as it involves Atlantic Forest, ideal conditions for the maintenance of the vector of the genus Amblyomma already described in the region and the presence of circulating Rickettsia rickettsii among dogs, confirmed by the presence of antibodies. Conducts regarding the awareness of the population through educational work should be implemented to prevent the disease in the population of the area.(AU)

HLA-B27 spondyloarthritis and spotted fever rickettsiosis: case-based review.

Rickettsia rickettsii, a tick borne disease, is the pathogen responsible for inducing Rocky Mountain Spotted Fever (RMSF), an illness that can progress to fulminant multiorgan failure and death. We present a case where R. rickettsii, acquired on a camping trip, precipitated a flare of peripheral arthritis and episcleritis in an HLA-B27 positive patient. Although Yersinia, Salmonella, Mycobacteria, Chlamydia, Shigella, Campylobacter, and Brucella have been previously associated with HLA-B27 spondyloarthritis, this unusual case demonstrates that obligate intracellular rickettsial organisms, and specifically, R. rickettsii, can also induce flares of HLA-B27 spondyloarthritis. Rickettsial infections in general can rapidly become fatal in both healthy and immunosuppressed patients, and thus, prompt diagnosis and therapy are required.

Development of an electrochemical immunosensor for the diagnostic testing of spotted fever using synthetic peptides.

Spotted fever is a rare acute and multisystemic febrile infectious disease with a mortality rate of ≥50% without adequate antibiotic treatment, and in diagnosed and treated cases, of approximately 2.5%. Currently, the applied test to diagnose this disease is the indirect immunofluorescence reaction, however two samples of paired sera are necessary to confirm the diagnosis, since using only one sample may allow for confusion with cross reactions. OmpA is an outer membrane protein present in the R. rickettsia, the etiological agent of spotted fever, able to activate dendritic and macrophage cells. It also presents immunogenicity properties, and is considered a target for the development of diagnostic tests for spotted fever. In this context, an amperometric immunosensor was developed for the identification of sera antibodies (human IgG) from patients with spotted fever aimed at improving sensitivity and minimize sample volume. The development of the immunosensor was conducted using a synthetic peptide, derivative from the H6PGA4 R. rickettsia protein, homologous to OmpA. Amperometric responses were generated at -0.6 to 0.6V, at a scan rate of 0.025Vs-1 for 20 cycles, a limit of detection of approximately 10ngmL-1 for the synthetic peptides and 0.01µgmL-1 for the humam serum, a sensitivity of 2.59µA, adequate for the detection of spotted fever antibodies. The construction of this immunosensor, capable of identifying circulating antibodies in real time, can also be applied in the diagnosis of other infectious-parasitic diseases.

Th1 epitope peptides induce protective immunity against Rickettsia rickettsii infection in C3H/HeN mice.

Rickettsia rickettsii is the causative pathogen of Rocky Mountain spotted fever (RMSF). Adr2, YbgF and OmpB are protective antigens of R. rickettsii. In this study, 90 candidate peptides were selected from these antigens based on their high-affinity binding capacity for the MHC class II molecule H2 I-A or H2 I-E using bioinformatic methods. Six peptides were determined using ELISPOT assay to be immunodominant based on the IFN-γ recall responses of CD4+ T cells from mice immunized with R. rickettsii. Six nucleotide sequences encoding the immunodominant peptides were linked in series and inserted into a plasmid for expression in Escherichia coli cells, resulting in a new, recombinant polypeptide termed GWP. After immunization and challenge, the rickettsial load or histopathological lesions in the organs of mice immunized with GWP or pooled peptides was significantly lower than that in organs of mice immunized with PBS or the individual peptide OmpB399. An in vitro neutralization test revealed that sera from mice immunized with GWP, OmpB399, or pooled peptides reduced R. rickettsii adherence to, and invasion of, vascular endothelial cells. Furthermore, significantly higher levels of IgG, IgG1, or IgG2a were detected in sera from mice immunized with GWP or pooled peptides, and significantly higher levels of IFN-γ or TNF-α secreted by CD4+ T cells from R. rickettsii-infected mice were detected after immunization with GWP. Altogether, our results indicated that polypeptides, especially GWP, could induce a Th1-type immune response against R. rickettsii infection, which might contribute to the rational design of peptide-based vaccines for RMSF.

Ehrlichia canis and Rickettsia spp. in dogs from urban areas in Paraiba state, northeastern Brazil / Ehrlichia canis e Rickettsia spp. em cães de áreas urbanas da Paraíba, nordeste do Brasil

Abstract The aims of our study was to identify Ehrlichia canis and antibodies against Rickettsia spp. belonging to the spotted fever group (SFG) in dogs sampled from Paraiba state, northeastern Brazil. Blood and serum samples collected by convenience from dogs in urban areas of five municipalities were analyzed by real-time PCR for the detection of E. canis DNA and by immunofluorescence assay test (IFAT) for the identification of antibodies against Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii and R. rhipicephali antigens. E. canis DNA was detected in 8.9% (64/719) of the blood samples, whereas 5.63% (43/763) of the serum samples were positive for at least one of the Rickettsia antigens tested by IFAT. This study showed for the first time the occurrence of E. canis and suggested the circulation of SFG Rickettsia in dogs in the study region of Paraiba state, northeastern Brazil.

Resumo Os objetivos do nosso estudo foram identificar Ehrlichia canis e anticorpos contra Rickettsia spp. pertencentes ao Grupo da Febre Maculosa (GFM) em cães amostrados no estado da Paraíba, nordeste do Brasil. As amostras de sangue e soro, coletados por conveniência, de cães em áreas urbanas de cinco municípios foram analisadas por PCR em tempo real para a detecção de DNA de E. canis e pela Reação de Imunofluorescência Indireta (RIFI) para identificação de anticorpos contra Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii e R. rhipicephali. O DNA de E. canis foi detectado em 8,9% (64/719) das amostras de sangue, enquanto que 5,63% (43/763) das amostras de soro foram positivas para pelo menos um dos antígenos de Rickettsia testados por RIFI. Este estudo mostrou pela primeira vez a ocorrência de E. canis e sugere a circulação de Rickettsia do GFM em cães na região em estudo do estado da Paraíba, Nordeste do Brasil.

Probability of occurrence of the Brazilian spotted fever in northeast of Paraná state, Brazil / Probabilidade de ocorrência de febre maculosa brasileira no norte pioneiro do estado do Paraná, Brasil

Abstract Brazilian spotted fever (BSF) is a fatal zoonosis because of the difficulties in its early diagnosis and treatment. Occurrences of BSF in the northeast of the state of Paraná prompted investigation of areas at risk of this rickettsiosis in the municipalities of Japira, Jaboti, Pinhalão and Tomazina. To determine the areas at risk, 592 serum samples from dogs and 230 from equids were analyzed by means of the indirect immunofluorescence assay (IFA) for Rickettsia rickettsii and R. parkeri . In addition, risk probability maps were drawn up using the kriging indicator technique. Among the samples tested, 5.3% (43/822) indicated presence of antibodies reactive to at least one of the two Rickettsia species tested: 7.8% of the equids (18/230) and 4.2% of the dogs (25/592) were positive. Geostatistical analysis showed that the average seropositivity rate was 5 to 6%. Although the average seropositivity rates observed among these dogs and equids were lower than those reported from endemic areas of Brazil, the biotic components (etiological agent, vector and reservoirs) and environmental aspects of BSF epidemiology were present in these municipalities.

Resumo A febre maculosa brasileira (FMB) é uma zoonose fatal devido às dificuldades para diagnosticá-la e tratá-la precocemente. A ocorrência de casos de FMB no Estado do Paraná suscitou a investigação de áreas de risco desta rickettsiose nos municípios de Japira, Jaboti, Pinhalão e Tomazina, na mesorregião norte pioneiro do Paraná. Para determinar as áreas de risco foram analisadas amostras de soro de 592 cães e 230 equídeos submetidos à reação de imunofluorescência indireta para Rickettsia rickettsii e R. parkeri. Além disto, foram construídos mapas de probabilidade de risco pela técnica de krigagem indicatriz. Das amostras testadas 5,3% (43/822) continham anticorpos para pelo menos uma das duas rickettsias testadas. Os equídeos apresentaram uma positividade de 7,8% (18/230) e os cães de 4,2% (25/592). A análise geoestatística mostrou que a soropositividade média é de 5 a 6%. Embora as soropositividade médias de cães e equídeos constatadas tenham sido menores do que as relatadas em áreas endêmicas do território brasileiro, os componentes bióticos (agente etiológico, vetor e reservatórios) e ambientais da epidemiologia da FMB se fazem presentes nos municípios referidos.

A fatal case of Brazilian spotted fever in a non-endemic area in Brazil: the importance of having health professionals who understand the disease and its areas of transmission

Abstract Brazilian spotted fever (BSF) is caused by the bacterium Rickettsia rickettsii. Because of its high case-fatality rate and apparent increase in areas of transmission, it is considered to be the rickettsial illness of primary public health interest. Cases of this disease have historically occurred in Southeastern Brazil. This article reports the first fatal case of BSF in Southern Brazil. This case high lights the importance of BSF to be considered as a differential diagnosis for acute hemorrhagic fever in areas where cases of BSF may not be expected.

Chloroform-Methanol Residue of Coxiella burnetii Markedly Potentiated the Specific Immunoprotection Elicited by a Recombinant Protein Fragment rOmpB-4 Derived from Outer Membrane Protein B of Rickettsia rickettsii in C3H/HeN Mice.

The obligate intracellular bacteria, Rickettsia rickettsii and Coxiella burnetii, are the potential agents of bio-warfare/bio-terrorism. Here C3H/HeN mice were immunized with a recombinant protein fragment rOmp-4 derived from outer membrane protein B, a major protective antigen of R. rickettsii, combined with chloroform-methanol residue (CMR) extracted from phase I C. burnetii organisms, a safer Q fever vaccine. These immunized mice had significantly higher levels of IgG1 and IgG2a to rOmpB-4 and interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), two crucial cytokines in resisting intracellular bacterial infection, as well as significantly lower rickettsial loads and slighter pathological lesions in organs after challenge with R. rickettsii, compared with mice immunized with rOmpB-4 or CMR alone. Additionally, after challenge with C. burnetii, the coxiella loads in the organs of these mice were significantly lower than those of mice immunized with rOmpB-4 alone. Our results prove that CMR could markedly potentiate enhance the rOmpB-4-specific immunoprotection by promoting specific and non-specific immunoresponses and the immunization with the protective antigen of R. rickettsii combined with CMR of C. burnetii could confer effective protection against infection of R. rickettsii or C. burnetii.

Surface protein Adr2 of Rickettsia rickettsii induced protective immunity against Rocky Mountain spotted fever in C3H/HeN mice.

BACKGROUND: Rickettsia rickettsii is the pathogen of Rocky Mountain spotted fever (RMSF), a life-threatening tick-transmitted infection. Adr2 was a surface-exposed adhesion protein of R. rickettsii and its immunoprotection against RMSF was investigated in mice. METHODS: Recombinant Adr2 (rAdr2) was used to immunize C3H/HeN mice, and the rickettsial loads in organs of the mice were detected after challenge with R. rickettsii. The levels of specific antibodies of sera from the immunized mice were determined and the sera from immunized mice were applied to neutralize R. rickettsii. Proliferation and cytokine secretion of CD4(+) and CD8(+) T cells isolated from R. rickettsii-infected mice were also assayed after rAdr2 stimulation. RESULTS: After R. rickettsii challenge, the rickettsial loads in spleens, livers, and lungs were significantly lower and the impairment degrees of these organs in rAdr2-immunized mice were markedly slighter, compared with those in negative control mice. The ratio of specific IgG2a/IgG1 of rAdr2-immunized mice kept increasing during the immunization. After treatment with rAdr2-immunized sera, the total number of R. rickettsii organisms adhering and invading host cells was significantly lower than that treated with PBS-immunized sera. Interferon-γ secretion by CD4(+) or CD8(+) T cells and tumor necrosis factor-α secretion by CD4(+) T cells from R. rickettsii-infected mice were respectively significantly greater than those from uninfected mice after rAdr2 stimulation. CONCLUSION: Adr2 is a protective antigen of R. rickettsii. Protection offered by Adr2 is mainly dependent on antigen-specific cell-mediated immune responses, including efficient activity of CD4(+) and CD8(+) T cells to produce great amount of TNF-α and/or IFN-γ as well as rapid increase of specific IgG2a, which synergistically activate and opsonize host cells to killing intracellular rickettsiae.

Serologic evidence of human Rickettsia infection found in three locations in Panamá / Evidencia serológica de infecciones de Rickettsia en humanos provenientes de tres localidades de Panamá

Introduction: Since the middle of last century, cases of rickettsiosis have been found in Panamá when outbreaks of murine typhus and spotted fever were reported. Since then, little information exists about its prevalence in this country, since it is most often is misdiagnosed as another disease. Objectives: The aim of this paper is to demonstrate the presence of Rickettsia infections in humans in three locations in Panamá. These locations are agricultural areas, near forested areas or those who work in zoo. Materials and methods: Three locations where chosen for this study: Tortí, El Valle de Antón and workers in the Summit Municipal Park in Panamá City. All volunteers signed an informed consent and answered a questionnaire. The samples were analyzed for the detection of rickettsial spotted fever and typhus group by the indirect immunofluorescence (using commercial kits) and antigens of Rickettsia rickettsii and R. amblyommii. Results: Blood samples were taken from 97 volunteers in Tortí (25), El Valle de Anton (37) and Summit Municipal Park (35). Of these, a total of 38 (39%) samples reacted to one of the two methods: eight (32%) in Tortí, 18 (48%) in El valle and 12 (34%) in Summit Municipal Park. Conclusion: The results show a high prevalence of antibodies to Rickettsia belonging to the spotted fever group in each of the three study areas, in addition to presenting evidence of the typhus group Rickettsia in El Valle de Anton. These areas could be considered endemic for rickettsiosis as there are conditions for maintaining them.

Introducción. Desde mediados del siglo pasado, se conocen en Panamá casos de rickettsiosis, cuando fueron reportados brotes de tifus en ratones y de fiebres manchadas. A partir de entonces, poca información se tiene sobre su prevalencia en este país, lo cual se debe principalmente a que son confundidos con otras enfermedades. Objetivos. El objetivo de este trabajo fue demostrar la presencia de rickettsiosis en humanos provenientes de tres localidades de Panamá, que corresponden a zonas agropecuarias, cercanas a bosques, o que trabajaban en zoológicos. Materiales y métodos. Se escogieron tres localidades para este estudio: Tortí (provincia de Panamá), El Valle de Antón (provincia de Coclé) y el Parque Municipal Summit en Ciudad de Panamá. Los voluntarios firmaron un consentimiento informado, además de responder un cuestionario. De cada voluntario se extrajo sangre venosa, la que fue analizada por medio de inmunoflorescencia indirecta, utilizando kits comerciales y láminas sensibilizadas con antígenos cultivados de Rickettsia rickettsii y Rickettsia amblyommii . Resultados. Se tomaron muestras de 97 voluntarios, 25 en Tortí, 37 en El Valle de Antón y 35 en el Parque Municipal Summit. De estos, 38 (39 %) de las muestras fueron positivas en algunas de las dos técnicas practicadas: 8 (32 %) en Tortí, 18 (48 %) en El Valle y 12 (34 %) en el Parque Municipal Summit. Conclusión. Se demuestra una alta prevalencia de anticuerpos contra Rickettsia del grupo de las fiebres manchadas en las tres áreas de estudio, además de presentarse evidencia de títulos para Rickettsia del grupo tifus en El Valle de Antón. Estas zonas podrían considerarse como endémicas por rickettsiosis, ya que existen condiciones que permiten el mantenimiento de las mismas.

Life-threatening ANCA-positive vasculitis associated with rickettsial infection.

We present a 47-year-old Caucasian fire fighter who developed multisystem organ failure in the setting of a positive antineutrophil cytoplasmic autoantibody (myeloperoxidase) as well as confirmed Rocky Mountain spotted fever by skin biopsy PCR. This case provided a diagnostic challenge, a rare association of a Rickettsia infection and autoimmune vasculitis as well as a unique management approach.

Fatal Brazilian spotless fever caused by Rickettsia rickettsii in a dark-skinned patient / Febre maculosa brasileira sem exantema causada por Rickettsia rickettsii em um paciente de cor negra

Brazilian spotted fever (BSF) is the most important and frequent rickettsial disease in Brazil. A fatal case of BSF is reported in a 32-year-old black man, who died of irreversible shock after five days of fever, severe headache and abdominal pain with no rash. Spleen, kidney and heart samples collected at autopsy were positive for Rickettsia rickettsii by PCR and sequencing. The authors emphasize the need for a high index of diagnostic suspicion for spotted fever in black patients. Absence of a skin rash should not dissuade clinicians from considering the possibility of BSF and initiating empirical therapy.

Febre maculosa brasileira (FMB) é a mais importante e frequente doença rickettsial no Brasil. Relatamos um caso fatal de FMB em um homem negro de 32 anos de idade que morreu de choque irreversível após cinco dias de febre, cefaléia intensa, dor abdominal, e sem evidência de exantema. Amostras de baço, rim e coração coletadas na necropsia foram positivas para Rickettsia rickettsii por PCR e sequenciamento. Os autores ressaltam a necessidade de um alto índice de suspeita diagnóstica para febre maculosa em pacientes negros. Ausência de exantema não deve dissuadir os clínicos de considerar a possibilidade de FMB e iniciar a terapêutica empírica.

Outbreak of Rocky Mountain spotted fever in Córdoba, Colombia

Rocky Mountain spotted fever (RMSF) is a tick-borne disease caused by the obligate intracellular bacterium Rickettsia rickettsii. Although RMSF was first reported in Colombia in 1937, it remains a neglected disease. Herein, we describe the investigation of a large cluster of cases of spotted fever rickettsiosis in a new area of Colombia.

Serological survey of Rickettsia sp. in horses and dogs in an non-endemic area in Brazil / Identificação sorológica de Rickettsia sp. em equinos e cães de área não endêmica no Brasil

Brazilian Spotted Fever (BSF) is a lethal rickettsiosis in humans caused by the bacteria Rickettsia rickettsii, and is endemic in some areas of Brazil. Horses and dogs are part of the disease's life cycle and they may also serve as sentinel animals in epidemiological studies. The first human BSF case in the State of Paraná was reported in 2005. The present study was conducted in the municipality of Almirante Tamandaré, where no previous case of BSF was reported. Serum samples were collected from 71 horses and 20 dogs from nine properties in the area. Ticks were also collected from these animals. All farmers completed a questionnaire about their knowledge of BSF and animal health management. Serum samples were analyzed by indirect immunofluorescent-antibody assay (IFA) using R. rickettsii and R. parkeri as antigens. Ticks were analyzed by PCR for Rickettsia sp., and all of them were PCR-negative. Six horses (8.45 percent) and 4 dogs (20 percent) were identified as seropositive. Farmers were not aware of the correlation between the presence of ticks and risk of BSF. Although a non-endemic area, Almirante Tamandaré is a vulnerable environment for BSF and effective tick control measures are required.

A Febre Maculosa Brasileira (FMB) é uma riquetsiose letal para humanos, causada pela bactéria Rickettsia rickettsii, e é endêmica em algumas regiões brasileiras. Equinos e cães podem participar do ciclo da doença e podem também servir como sentinelas em estudos epidemiológicos. O primeiro caso humano relatado no Estado do Paraná ocorreu em 2005. O presente estudo foi realizado no município de Almirante Tamandaré, região onde não há relatos de casos de FMB. Foram coletadas amostras de sangue de 71 cavalos e 20 cães em nove propriedades rurais na região. Carrapatos também foram colhidos dos animais. Todos os proprietários responderam a um questionário sobre o manejo sanitário dos animais e o conhecimento a respeito da FMB. As amostras de soro foram processadas pela técnica de Reação de Imunofluorescência Indireta (RIFI), utilizando-se os antígenos de R. rickettsii e R. parkeri. Os carrapatos foram analisados por PCR para Rickettsia sp. e todos foram negativos. Seis cavalos (8,45 por cento) e 4 cães (20 por cento) foram identificados como soropositivos. Todos os proprietários desconheciam a relação de carrapatos com a FMB. Embora considerada uma área não endêmica, Almirante Tamandaré é um ambiente vulnerável à FMB e um controle eficiente de carrapatos deve ser implementado.

The expanding spectrum of eschar-associated rickettsioses in the United States.

BACKGROUND: Until recently, Rickettsia rickettsii was the only substantiated cause of tick-borne spotted fever group (SFG) rickettsiosis in humans in the United States. Rickettsia parkeri, originally thought to be nonpathogenic in humans, was recently proved to be another cause of tick-borne SFG rickettsiosis. OBSERVATIONS: We report 3 cases of SFG rickettsiosis and discuss the epidemiology, clinical presentation, histopathologic features, and laboratory findings that support confirmed or probable diagnoses of R parkeri infection and describe the expanding list of eschar-associated SFG rickettsioses recognized in US patients. CONCLUSIONS: The SFG rickettsioses share many clinical manifestations and extensive antigenic cross-reactivity that may hamper specific confirmation of the causative agent.

Prevalence of Rickettsia species antibodies and Rickettsia species DNA in the blood of cats with and without fever.

Rickettsia species antibodies have been detected in some cats but it is unknown whether infected cats develop clinical signs. The prevalence of Rickettsia species deoxyribonucleic acid (DNA) in blood from clinically ill cats has not been determined. The objective of this study was to determine if cats with fever (body temperature >or=102.5 degrees F [39.2 degrees C]) were more likely to have evidence of rickettsial infection than healthy, age-matched, control cats with a body temperature<102.5 degrees F. Rickettsia species polymerase chain reaction (PCR) assays were performed to detect rickettsial DNA extracted from blood (71 paired samples), indirect immunofluorescence assays (IFA) were performed to detect serum antibodies against Rickettsia felis (90 paired samples) and Rickettsia rickettsii (91 paired samples), and the results between pairs were compared. All samples were negative for Rickettsia species DNA. More cats with fever were seropositive for R felis or R rickettsii than control cats, but results were not statistically significant. Results of this pilot study failed to show an association between Rickettsia species DNA or Rickettsia species antibodies and fever.

Characterization of rickettsia rickettsii in a case of Fatal Brazilian spotted fever in the city of Rio de Janeiro, Brazil

A lethal case of Brazilian spotted fever (BSF) is presented. Clinical features were initially of gastrointestinal involvement and evolved with progression to septic shock, meningoencephalitis and death on the 6th day of illness. Indirect immunofluorescence assay (IFA) for spotted fever group rickettsia (SFGR) was non-reactive. Diagnosis was confirmed by the polymerase chain reaction (PCR) and the nucleotide sequencing of a fragment of the ompA gene showed 100 percent homology to Rickettsia rickettsii. BSF has not been reported in the city of Rio de Janeiro in the last three decades, and the present description should alert the clinicians to its presence in urban Rio de Janeiro, and to the differential diagnosis with dengue fever, gastroenteritis, leptospirosis and bacterial septic shock, among others.

Host defenses to Rickettsia rickettsii infection contribute to increased microvascular permeability in human cerebral endothelial cells.

Rickettsiae are arthropod-borne intracellular bacterial pathogens that primarily infect the microvascular endothelium leading to systemic spread of the organisms and the major pathophysiological effect, increased microvascular permeability, and edema in vital organs such as the lung and brain. Much work has been done on mechanisms of immunity to rickettsiae, as well as the responses of endothelial cells to rickettsial invasion. However, to date, no one has described the mechanisms of increased microvascular permeability during acute rickettsiosis. We sought to establish an in vitro model of human endothelial-target rickettsial infection using the etiological agent of Rocky Mountain spotted fever, Rickettsia rickettsii, and human cerebral microvascular endothelial cells. Endothelial cells infected with R. rickettsii exhibited a dose-dependent decrease in trans-endothelial electrical resistance, which translates into increased monolayer permeability. Additionally, we showed that the addition of pro-inflammatory stimuli essential to rickettsial immunity dramatically enhanced this effect. This increase in permeability correlates with dissociation of adherens junctions between endothelial cells and is not dependent on the presence of nitric oxide. Taken together, these results demonstrate for the first time that increased microvascular permeability associated with rickettsial infection is partly attributable to intracellular rickettsiae and partly attributable to the immune defenses that have evolved to protect the host from rickettsial spread.

Rickettsia rickettsii infection of cultured human endothelial cells induces heme oxygenase 1 expression.

Existing evidence suggests that oxidative insults and antioxidant defense mechanisms play a critical role in the host cell response during infection of endothelial cells by Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever. Heme oxygenase (HO), a rate-limiting enzyme in the pathway for heme catabolism, protects against oxidant damage in a variety of stress situations. Here, we report on the expression of the inducible and constitutive HO isozymes, HO-1 and HO-2, during R. rickettsii infection of endothelial cells. Steady-state levels for HO-1 mRNA were increased two- to threefold, as early as 4 h postinfection, whereas HO-2 mRNA was not affected. Induction of HO-1 mRNA was dependent on the dose of infection and occurred in a time-dependent manner, reaching maximal levels at 4 to 7 h. The increase in HO-1 mRNA occurred at the level of trancription as it was blocked by the transcriptional inhibitors, actinomycin D and alpha-amanitin. The eukaryotic protein synthesis inhibitor, cycloheximide, caused a >50% reduction in the infection-induced increase in HO-1 mRNA level, suggesting its dependence on de novo protein synthesis of host cell. The uptake of viable organisms appeared to be necessary, since inactivation of R. rickettsii by heat or formalin fixation, or incubation of cells with cytochalasin B to prevent entry resulted in marked inhibition of HO-1 response. N-Acetyl-L-cysteine, a known oxidant scavenger, inhibited the HO-1 induction by R. rickettsii. Finally, Western analysis with a specific monoclonal antibody revealed higher levels of HO-1 protein ( approximately 32 kDa), confirming that changes in HO-1 mRNA levels were followed by increases in the levels of protein. The findings indicate that R. rickettsii infection induces HO-1 expression in host endothelial cells and suggest an important role for this enzyme in cellular response to infection, possibly by serving a protective function against oxidative injury.

Evidence of rickettsial spotted fever and ehrlichial infections in a subtropical territory of Jujuy, Argentina.

Between November 1993 and March 1994, a cluster 6 pediatric patients with acute febrile illnesses associated with rashes was identified in Jujuy Province, Argentina. Immunohistochemical staining of tissues confirmed spotted fever group rickettsial infection in a patient with fatal disease, and testing of serum of a patient convalescing from the illness by using an indirect immunofluorescence assay (IFA) demonstrated antibodies reactive with spotted fever group rickettsiae. A serosurvey was conducted among 16 households in proximity to the index case. Of 105 healthy subjects evaluated by IFA, 19 (18%) demonstrated antibodies reactive with rickettsiae or ehrlichiae: 4 had antibodies reactive with Rickettsia rickettsii, 15 with Ehrlichia chaffeensis, and 1 with R. typhi. Amblyomma cajennense, a known vector of R. rickettsii in South America, was collected from pets and horses in the area. These results are the first to document rickettsial spotted fever and ehrlichial infections in Argentina.