Serologic evidence of rickettsial diseases associated with tick bites in workers of urban veterinary clinics.

BACKGROUND & OBJECTIVES: Rickettsial and other zoonotic diseases are a latent risk for workers of veterinary clinics. The objective of this study was to evaluate the prevalence and the associated risk factors of parasitosis caused by Rhipicephalus sanguineus, and to estimate the seroprevalence of rickettsial diseases in workers of urban veterinary clinics of Juárez city, México. METHODS: The participants of the study were recruited from 63 private veterinary clinics and hospitals. The serological analysis of the blood samples collected was carried out using immunofluorescence assay (IFA). The statistical analysis for prevalences, risk factors, and correlation was performed with the SAS program. RESULTS: In total, 167 veterinary workers were included in the study. The prevalence of tick bites was 40% (67/167), and the risk factors associated with the occurrence of bites included the activities performed in the clinic and the number of labour hours spent per week. About 21% (35/167) of participants were seropositive to R. rickettsii, 28% (47/167) to Ehrlichia chaffeensis, and 24% (40/167) to Anaplasma phagocytophilum. A correlation was observed between: the number of workers in the clinics and the proportion of tick bites (r2 = 0.865); the prevalence of bites and the seropositivity of the participants to at least one pathogen (r2 = 0.924); and the number of bites per individual and infection to pathogens (r2 = 0.838). INTERPRETATION & CONCLUSION: Workers in urban veterinary clinics are highly exposed to tick bites and, therefore, to the diseases they transmit. Hence, it is important to implement prevention measures and perform constant monitoring of these diseases.

The rice rat Euryoryzomys russatus, a competent amplifying host of Rickettsia parkeri strain Atlantic rainforest for the tick Amblyomma ovale.

Rickettsia parkeri strain Atlantic rainforest (SAR) is the etiological agent of a spotted fever group rickettsiosis in Brazil, where it is transmitted to humans by the tick Amblyomma ovale. A previous study demonstrated that R. parkeri SAR was successfully maintained in A. ovale ticks by transstadial and transovarial passages; however, because this agent induced lower reproduction rates in A. ovale, the participation of a vertebrate amplifier host, yet to be determined, was speculated. Since the rice rat Euryoryzomys russatus was demonstrated to be the most important host for immature stages of A. ovale in a focus of R. parkeri SAR transmission, the present study evaluated the competence of rice rats to act as amplifying host of R. parkeri SAR for A. ovale ticks. Rice rats were infested with R. parkeri SAR-infected A. ovale nymphs, and four days later with uninfected A. ovale larvae. Rickettsial transmission to rats was confirmed by seroconversion to R. parkeri antigens. Detached engorged larvae were allowed to molt to nymphs, in which rickettsial DNA was detected in up to 60% (mean: 20%) of the specimens. When part of these nymphs was allowed to feed on susceptible rice rats, rickettsial transmission was confirmed by seroconversion, indicating that there was successful horizontal transmission of R. parkeri SAR from infected nymphs to uninfected larvae in the previous acquisition infestations. Because we used naïve, susceptible rats, we infer that this horizontal transmission occurred via a systemic infection (rickettsemia) in the rat. Our results, coupled with previous epidemiological studies, suggest that under natural conditions rice rats could be acting as amplifying hosts of R. parkeri SAR to A. ovale ticks.

Eco-epidemiological analysis of rickettsial seropositivity in rural areas of Colombia: A multilevel approach.

Rickettsiosis is a re-emergent infectious disease without epidemiological surveillance in Colombia. This disease is generally undiagnosed and several deadly outbreaks have been reported in the country in the last decade. The aim of this study is to analyze the eco-epidemiological aspects of rickettsial seropositivity in rural areas of Colombia where outbreaks of the disease were previously reported. A cross-sectional study, which included 597 people living in 246 households from nine hamlets in two municipalities of Colombia, was conducted from November 2015 to January 2016. The survey was conducted to collect sociodemographic and household characteristics (exposure) data. Blood samples were collected to determine the rickettsial seropositivity in humans, horses and dogs (IFA, cut-off = 1/128). In addition, infections by rickettsiae were detected in ticks from humans and animals by real-time PCR targeting gltA and ompA genes. Data was analyzed by weighted multilevel clog-log regression model using three levels (person, household and hamlets) and rickettsial seropositivity in humans was the main outcome. Overall prevalence of rickettsial seropositivity in humans was 25.62% (95%CI 22.11-29.12). Age in years (PR = 1.01 95%CI 1.01-1.02) and male sex (PR = 1.65 95%CI 1.43-1.90) were risk markers for rickettsial seropositivity. Working outdoors (PR = 1.20 95%CI 1.02-1.41), deforestation and forest fragmentation for agriculture use (PR = 1.75 95%CI 1.51-2.02), opossum in peridomiciliary area (PR = 1.56 95%CI 1.37-1.79) and a high proportion of seropositive domestic animals in the home (PR20-40% vs <20% = 2.28 95%CI 1.59-3.23 and PR>40% vs <20% = 3.14 95%CI 2.43-4.04) were associated with rickettsial seropositivity in humans. This study showed the presence of Rickettsia antibodies in human populations and domestic animals. In addition, different species of rickettsiae were detected in ticks collected from humans and animals. Our results highlighted the role of domestic animals as sentinels of rickettsial infection to identify areas at risk of transmission, and the importance of preventive measures aimed at curtailing deforestation and the fragmentation of forests as a way of reducing the risk of transmission of emergent and re-emergent pathogens.

Detecção de proteínas imunorreativas de Rickettsia sp. cepa Mata Atlântica / Detection of immunoreactive proteins of Rickettsia sp. Atlantic Forest strain

A Febre Maculosa Brasileira (FMB) é uma doença infecciosa, transmitida por carrapatos ao homem. Uma nova riquetsiose humana foi descrita como causadora de Febre Maculosa no Estado de São Paulo, sendo denominada de Rickettsia sp. cepa Mata Atlântica. O presente trabalho teve como objetivo detectar e identificar proteínas com potencial de estimular o sistema imune de hospedeiro mamífero, desta nova cepa descrita. Para tanto, foi realizado a extração proteica total de Rickettsia sp. cepa Mata Atlântica. As proteínas extraídas foram fracionadas por eletroforese. As bandas proteicas foram transferidas para membranas de nitrocelulose por migração elétrica e submetidas à técnica de Western-blot, para detecção proteica. Ao todo sete proteínas imunorreativas foram detectadas. Duas proteínas apresentaram maior abundancia, com peso molecular, de 200 e 130 kDa respectivamente. Através da comparação de mapas proteômicos existentes e pelo peso molecular que estas proteínas apresentaram, sugere-se que as duas proteínas detectadas representem rOmpA (200 kDa) e rOmpB (130 kDa). As demais proteínas detectadas apresentaram menor ocorrência e peso molecular inferior a 78 kDa, podendo representar membros da família de antígenos de superfície celular (Sca - Surface cell antigen). As proteínas detectadas poderão servir como base de estudo na elaboração de métodos diagnósticos sensíveis e específicos, no desenvolvimento de vacinas, além de possibilitarem novos estudos para terapias mais eficazes.(AU)

Brazilian Spotted Fever (BSF) is an infectious disease transmitted by ticks to humans. A new human rickettsial infection was reported to cause spotted fever in the State of São Paulo and was named Rickettsia sp. Strain Atlantic Forest. This study aimed to detect and identify proteins with potential to stimulate the immune system of mammalian host of this new strain described. Therefore, we performed total protein extraction Rickettsia sp. Strain Atlantic Forest. The extracted proteins were fractionated by electrophoresis. The protein bands were transferred to nitrocellulose membrane by electrical migration and subjected to Western blot for protein detection. In all, seven immunoreactive proteins were detected. Two proteins showed higher abundance, with molecular weight of 200 and 130 kDa respectively. By comparing existing proteomic maps and the molecular weight of these proteins showed that, it is suggested that the two proteins detected representing rOmpA (200 kDa) and rOmpB (130 kDa). The other detected proteins had lower occurrence and molecular weight less than 78 kDa, which may represent members of the cell surface antigens Family (Sca - Surface cell antigen). The detected proteins may serve as a study based on the development of sensitive and specific diagnostic methods in the development of vaccines and they enable further studies to more effective therapies.(AU)

First Identification and Description of Rickettsioses and Q Fever as Causes of Acute Febrile Illness in Nicaragua.

BACKGROUND: Rickettsial infections and Q fever present similarly to other acute febrile illnesses, but are infrequently diagnosed because of limited diagnostic tools. Despite sporadic reports, rickettsial infections and Q fever have not been prospectively studied in Central America. METHODOLOGY/PRINCIPAL FINDINGS: We enrolled consecutive patients presenting with undifferentiated fever in western Nicaragua and collected epidemiologic and clinical data and acute and convalescent sera. We used ELISA for screening and paired sera to confirm acute (≥4-fold rise in titer) spotted fever and typhus group rickettsial infections and Q fever as well as past (stable titer) infections. Characteristics associated with both acute and past infection were assessed. CONCLUSIONS/SIGNIFICANCE: We enrolled 825 patients and identified acute rickettsial infections and acute Q fever in 0.9% and 1.3%, respectively. Clinical features were non-specific and neither rickettsial infections nor Q fever were considered or treated. Further study is warranted to define the burden of these infections in Central America.

Serological evidence of Rickettsia and Coxiella burnetii in humans of Buenos Aires, Argentina.

In Buenos Aires city (Argentina), the circulation of these agents has been detected mainly in vectors and animals, few human cases having been described. The aim of our study was to determine the seroprevalence of Rickettsia (spotted fever--SFG--and typhus--TG--groups) and Coxiella burnetii (Q fever agent) in residents of Buenos Aires city. The study involved 99 participants. Rickettsia IgG antibodies against SFG and TG were detected by IFA in 28.3% and 16.2% of serum samples, respectively. SFG titers were mostly 1/64 (53.6%) with a maximum of 1/512 (3.5%) whereas TG titers ranged between 1/64 (62.5%) and 1/256 (6.3%). Only one sample showed a titer of 1/32 for C. burnetii (phases I and II). The circulation of these pathogens in urban areas such as the city of Buenos Aires should be considered by health services, especially at the primary care level.

Pathogenic potential of a Costa Rican strain of 'Candidatus Rickettsia amblyommii' in guinea pigs (Cavia porcellus) and protective immunity against Rickettsia rickettsii.

'Candidatus Rickettsia amblyommii' is a spotted fever group rickettsia that is not considered pathogenic, although there is serologic evidence of possible infection in animals and humans. The aim of this study was to evaluate the pathogenic potential of a Costa Rican strain of 'Candidatus R. amblyommii' in guinea pigs and determine its capacity to generate protective immunity against a subsequent infection with a local strain of Rickettsia rickettsii isolated from a human case. Six guinea pigs were inoculated with 'Candidatus R. amblyommii' strain 9-CC-3-1 and two controls with cell culture medium. Health status was evaluated, and necropsies were executed at days 2, 4, and 13. Blood and tissues were processed by PCR to detect the gltA gene, and end titers of anti-'Candidatus R. amblyommii' IgG were determined by indirect immunofluorescence. To evaluate protective immunity, another 5 guinea pigs were infected with 'Candidatus R. amblyommii' (IGPs). After 4 weeks, these 5 IGPs and 3 controls (CGPs) were inoculated with pathogenic R. rickettsii. Clinical signs and titers of anti-Rickettsia IgG were determined. IgG titers reached 1:512 at day 13 post-infection with 'Candidatus R. amblyommii'. On day 2 after inoculation, two guinea pigs had enlarged testicles and 'Candidatus R. amblyommii' DNA was detected in testicles. Histopathology confirmed piogranulomatous orchitis with perivascular inflammatory infiltrate in the epididymis. In the protective immunity assay, anti-Rickettsia IgG end titers after R. rickettsii infection were lower in IGPs than in CGPs. IGPs exhibited only transient fever, while CGP showed signs of severe disease and mortality. R. rickettsii was detected in testicles and blood of CGPs. Results show that the strain 9-CC-3-1 of 'Candidatus R. amblyommii' was able to generate pathology and an antibody response in guinea pigs. Moreover, its capacity to generate protective immunity against R. rickettsii may modulate the epidemiology and severity of Rocky Mountain spotted fever in areas where both species circulate.

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.

Serologic evidence of human Rickettsia infection found in three locations in 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.

Anticorpos anti-Rickettsia spp. em capivaras de vida livre e de cativeiro no Sul do Brasil / Anti-Rickettsia spp. antibodies in free-ranging and captive capybaras from southern Brazil

Capybaras (Hydrochaeris hydrochaeris) are among the main hosts of Amblyomma spp. ticks, which is able to transmit Rickettsia species to human beings and animals. Since they are often infested with potential vector ticks, capybaras may be used as sentinels for rickettsiosis, such as the Brazilian Spotted Fever. The aim of the present study was to determine the prevalence of antibodies against Rickettsia spp. using the indirect immunofluorescence assay (IFA) in 21 free-ranging and 10 captive animals from the Zoological Park of the 'Bela Vista Biological Sanctuary' (BVBS), Itaipu Binational, Foz do Iguaçu, Southern Brazil. Antigens of six rickettsial species already identified in Brazil (Rickettsia rickettsii, R. parkeri, R. bellii, R. rhipicephali, R. amblyommii and R. felis) were used for IFA. Ticks from each capybara were collected for posterior taxonomic identification. A total of 19 (61.3 percent) samples reacted to at least one of tested species. Seropositivity was found in 14 (45.2 percent), 12 (38.7 percent), 5 (16.1 percent), 4 (12.9 percent), 3 (9.7 percent) and 3 (9.7 percent) animals for R. rickettsii, R. bellii, R. parkeri, R. amblyommii, R. felis and R. rhipicephali, respectively. Two captive capybaras presented suggestive titers of R. rickettsii infection and one sample showed homologous reaction to R. parkeri. Only one free-ranging capybara presented evidence R. bellii infection. Ticks collected on capybaras were identified as Amblyomma dubitatum e Amblyomma sp. Results evidenced the rickettsial circulation in the area, suggesting a potential role of capybaras on bacterial life cycle.(AU)

As capivaras (Hydrochaeris hydrochaeris) estão entre os principais hospedeiros do carrapato Amblyomma spp., o qual pode transmitir algumas espécies de riquétsias para seres humanos e animais. Como são frequentemente infestadas por carrapatos vetores potenciais, as capivaras podem ser usadas como sentinelas para riquetsioses, como a Febre Maculosa Brasileira. O objetivo do presente estudo foi determinar a soroprevalência de Rickettsia spp. por meio da reação de Imunofluorescência Indireta (RIFI) em 21 capivaras de vida livre e 10 capivaras de cativeiro do Zoológico do Refúgio Biológico Bela Vista, Itaipu Binacional, Foz do Iguaçu, Brasil. Antígenos de seis espécies de riquétsias já identi[1]icadas no Brasil (Rickettsia rickettsii, R. parkeri, R. bellii, R. rhipicephali, R. amblyommii e R. felis) foram utilizados para a RIFI. Carrapatos de cada capivara foram coletados para posterior identi[1]icação taxonômica. Um total de 19 (61,3 por cento) amostras reagiu a pelo menos uma das espécies testadas. Foi encontrada soropositividade em 14 (45,2 por cento), 12 (38,7 por cento), 5 (16,1 por cento), 4 (12,9 por cento), 3 (9,7 por cento) e 3 (9,7 por cento) animais para R. rickettsii, R. bellii, R. parkeri, R. amblyommii, R. felis e R. rhipicephali, respectivamente. Duas capivaras de cativeiro apresentaram títulos sugestivos de infecção por R. rickettsii e uma amostra apresentou reação homóloga frente à R. parkeri. Apenas uma capivara de vida livre apresentou evidência de infecção por R. bellii. Os carrapatos coletados sobre as capivaras foram identificados como Amblyomma dubitatum e Amblyomma sp. Os resultados evidenciam a circulação de riquétsias na região, sugerindo uma potencial participação da capivara no ciclo de vida desta bactéria.(AU)

[Spotted fever Rickettsiae. Their immunodominant proteins and immune response]. / Las Rickettsias del grupo de las fiebres manchadas: respuesta inmune y sus proteínas inmunodominantes.

The organisms of Rickettsia species are gram (-) bacteria that cause severe illnesses in humans and are an important health problem in several countries around the world, including Mexico. The presence of different antigens between species and also in different strains of the same specie is an obstacle for vaccine development and serological diagnosis. There are important advances in the knowledge of the rickettsial antigenic structure and the resulting immune response in infected hosts, including humans. This review covers these topics and provides an overview about the development of vaccines and accessible diagnostic methods for diseases caused by Rickettsia.

Las Rickettsias del grupo de las fiebres manchadas: respuesta inmune y sus proteínas inmunodominantes / Spotted fever Rickettsiae: their immunodominant proteins and immune response

The organisms of Rickettsia species are Gram (-) bacteria that cause severe illnesses in humans and are an important health problem in several countries around the world, including Mexico. The presence of different antigens between species and also in different strains of the same specie is an obstacle for vaccine development and serological diagnosis. There are important advances in the knowledge of the rickettsial antigenic structure and the resulting immune response in infected hosts, including humans. This review covers these topics and provides an overview about the development of vaccines and accessible diagnostic methods for diseases caused by Rickettsia (Rev Méd Chile 2004; 132: 381-7).

An apparent outbreak of rickettsial illness in Costa Rica, 1974.

A focal outbreak of highly fatal disease occurred in eastern Costa Rica in 1974. No rickettsial spotted fevers have previously been reported in Costa Rica. Nevertheless, the available evidence points to an unidentified rickettsial pathogen, probably of the spotted fever group, as the agent responsible for this outbreak.