RESUMEN
BACKGROUND: The tick Amblyomma sculptum is the major vector of Rickettsia rickettsii, the causative agent of the highly lethal Brazilian spotted fever. It has been shown that R. rickettsii inhibits apoptosis in both human endothelial cells and tick cells. Apoptosis is regulated by different factors, among which inhibitors of apoptosis proteins (IAPs) play a central role. In the study reported here, we selected an IAP of A. sculptum that has not yet been characterized to assess its role in cell death and to determine the effects of its gene silencing on tick fitness and R. rickettsii infection. METHODS: An A. sculptum cell line (IBU/ASE-16) was treated with specific double-stranded RNA (dsRNA) for either IAP (dsIAP) or green fluorescent protein (dsGFP; as a control). The activity of caspase-3 and the exposure of phosphatidylserine were determined in both groups. In addition, unfed adult ticks, infected or not infected with R. rickettsii, were treated with either dsIAP or dsGFP and allowed to feed on noninfected rabbits. In parallel, noninfected ticks were allowed to feed on an R. rickettsii-infected rabbit. Ticks (infected or not with R. rickettsii) that remained unfed were used as a control. RESULTS: Caspase-3 activity and the externalization of phosphatidylserine were significantly higher in IBU/ASE-16 cells treated with dsIAP than in those treated with dsGFP. The mortality rates of ticks in the dsIAP group were much higher than those in the dsGFP group when they were allowed to feed on rabbits, independent of the presence of R. rickettsii. Conversely, lower mortality rates were recorded in unfed ticks. CONCLUSIONS: Our results show that IAP negatively regulates apoptosis in A. sculptum cells. Moreover, IAP-silenced ticks experienced higher mortality rates following the acquisition of a blood meal, suggesting that feeding may trigger the activation of apoptosis in the absence of this physiological regulator. These findings indicate that IAP is a potential antigen for an anti-tick vaccine.
Asunto(s)
Ixodidae , Fiebre Maculosa de las Montañas Rocosas , Garrapatas , Animales , Humanos , Conejos , Garrapatas/microbiología , Amblyomma , Caspasa 3/metabolismo , Ixodidae/genética , Proteínas Inhibidoras de la Apoptosis/metabolismo , Células Endoteliales , Fosfatidilserinas/metabolismo , Rickettsia rickettsii/fisiología , BrasilRESUMEN
Brazilian spotted fever (BSF) is an acute infectious disease caused by the bacterium Rickettsia rickettsii, which is transmitted by different tick species. Due to deleterious effects caused on ticks, the horizontal transmission of R. rickettsii through amplifying hosts is crucial for its maintenance in tick populations among BSF-endemic areas. The tick Amblyomma aureolatum is the main vector of R. rickettsii in the São Paulo metropolitan area; nevertheless, it is not known which vertebrate could act as an amplifying host for this tick species. Herein, we evaluated the potential of domestic dogs - primary hosts for A. aureolatum adults in BSF-endemic areas - to act as amplifying hosts. For this purpose, A. aureolatum non-infected adults were allowed to feed on two groups of dogs: the control group (G1), composed of one dog not exposed to R. rickettsii; and, the infected group (G2), composed of three dogs infected with R. rickettsii via tick parasitism. All G2-dogs became ill, seroconverted to R. rickettsii, and rickettsial DNA was detected in 87% of the engorged females that fed on them. Transovarial transmission rate was estimated to be 25% and infected larvae successfully transmitted R. rickettsii to guinea-pigs, confirming transovarial transmission and vector competence. No rickettsial DNA was detected in individual samples of eggs or larvae, which precluded the estimation of filial infection rate, but implies that it was low. Our results suggest that domestic dogs act as amplifying hosts of R. rickettsii for A. aureolatum ticks in BSF-endemic areas in Brazil.
Asunto(s)
Amblyomma/microbiología , Zoonosis Bacterianas/transmisión , Enfermedades de los Perros/transmisión , Interacciones Huésped-Patógeno , Infecciones por Rickettsia/veterinaria , Rickettsia rickettsii/fisiología , Animales , Zoonosis Bacterianas/microbiología , Brasil , Enfermedades de los Perros/microbiología , Perros , Femenino , Larva/crecimiento & desarrollo , Larva/fisiología , Masculino , Ninfa/crecimiento & desarrollo , Ninfa/fisiología , Infecciones por Rickettsia/microbiología , Infecciones por Rickettsia/transmisiónRESUMEN
BACKGROUND: Rickettsia rickettsii is a tick-borne obligate intracellular bacterium that causes Rocky Mountain spotted fever, a life-threatening illness. To obtain an insight into the vector-pathogen interactions, we assessed the effects of infection with R. rickettsii on the proteome cells of the tick embryonic cell line BME26. METHODS: The proteome of BME26 cells was determined by label-free high-performance liquid chromatography coupled with tandem mass spectrometry analysis. Also evaluated were the effects of infection on the activity of caspase-3, assessed by the hydrolysis of a synthetic fluorogenic substrate in enzymatic assays, and on the exposition of phosphatidyserine, evaluated by live-cell fluorescence microscopy after labeling with annexin-V. Finally, the effects of activation or inhibition of caspase-3 activity on the growth of R. rickettsii in BME26 cells was determined. RESULTS: Tick proteins of different functional classes were modulated in a time-dependent manner by R. rickettsii infection. Regarding proteins involved in apoptosis, certain negative regulators were downregulated at the initial phase of the infection (6 h) but upregulated in the middle of the exponential phase of the bacterial growth (48 h). Microorganisms are known to be able to inhibit apoptosis of the host cell to ensure their survival and proliferation. We therefore evaluated the effects of infection on classic features of apoptotic cells and observed DNA fragmentation exclusively in noninfected cells. Moreover, both caspase-3 activity and phosphatidylserine exposition were lower in infected than in noninfected cells. Importantly, while the activation of caspase-3 exerted a detrimental effect on rickettsial proliferation, its inhibition increased bacterial growth. CONCLUSIONS: Taken together, these results show that R. rickettsii modulates the proteome and exerts an inhibitory effect on apoptosis in tick cellsthat seems to be important to ensure cell colonization.
Asunto(s)
Apoptosis , Rickettsia rickettsii/fisiología , Garrapatas/citología , Garrapatas/microbiología , Animales , Caspasa 3/genética , Caspasa 3/metabolismo , Interacciones Huésped-Patógeno , Garrapatas/genética , Garrapatas/metabolismoRESUMEN
Although the ticks Amblyomma sculptum and Amblyomma aureolatum are important vectors of Rickettsia rickettsii, causative agent of the life-threatening Rocky Mountain spotted fever, A. aureolatum is considerably more susceptible to infection than A. sculptum. As the microbiota can interfere with the colonization of arthropod midgut (MG) by pathogens, in the current study we analyzed the MG microbiota of both tick species. Our results revealed that the MG of A. aureolatum harbors a prominent microbiota, while A. sculptum does not. Remarkably, a significant reduction of the bacterial load was recorded in R. rickettsii-infected A. aureolatum. In addition, the taxonomy analysis of the MG bacterial community of A. aureolatum revealed a dominance of the genus Francisella, suggesting an endosymbiosis. This study is the first step in getting insights into the mechanisms underlying the interactions among Amblyomma species, their microbiota and R. rickettsii. Additional studies to better understand these mechanisms are required and may help the development of novel alternatives to block rickettsial transmission.
Asunto(s)
Amblyomma/microbiología , Vectores Arácnidos/microbiología , Francisella/fisiología , Tracto Gastrointestinal/microbiología , Microbiota/fisiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Animales , Susceptibilidad a Enfermedades , Vectores de Enfermedades , Interacciones Huésped-Patógeno , Humanos , SimbiosisRESUMEN
Brazilian spotted fever (BSF), caused by the bacterium Rickettsia rickettsii, is transmitted to humans mainly by the tick Amblyomma sculptum in southeastern Brazil. In most BSF-endemic areas A. sculptum populations are sustained chiefly by capybaras (Hydrochoerus hydrochaeris), which are also the host of another tick species, Amblyomma dubitatum, not implicated in R. rickettsii transmission. Herein, we evaluated the effects of relative humidity (RH), temperature, and water immersion on the successful development of the engorged stages of A. dubitatum and compared them with recently published data under the same experimental conditions for A. sculptum. We showed that free-living developmental stages (engorged larvae, nymphs and females, and eggs) of A. dubitatum had higher survival rates when these stages were submerged in water for different periods of times (24-72â¯h). On the other hand, higher survival rates were observed for A. sculptum when ticks were incubated at lower RH values, 65 or 78 %, regardless of summer or winter mean temperatures. These results suggest that A. dubitatum is more adapted to humid habitats and seasonally flooding soils than A. sculptum, whereas this latter species should be more adapted than A. dubitatum to drier habitats. The implications of these results for the spatial distribution of A. dubitatum and A. sculptum, in relation to BSF epidemiology, are discussed.
Asunto(s)
Amblyomma/fisiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/epidemiología , Amblyomma/crecimiento & desarrollo , Animales , Brasil/epidemiología , Femenino , Inmersión , Laboratorios , Larva/crecimiento & desarrollo , Larva/fisiología , Ninfa/crecimiento & desarrollo , Ninfa/fisiología , Óvulo/crecimiento & desarrollo , Óvulo/fisiología , Roedores/microbiología , Estaciones del Año , Especificidad de la EspecieRESUMEN
Amblyomma aureolatum ticks are vectors of Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever in Brazil. Maintenance of R. rickettsii in nature depends on horizontal transmission along tick generations. Although such transmission is known to occur when uninfected and infected ticks feed simultaneously on susceptible animals (co-feeding systemic transmission), we investigated co-feeding nonsystemic transmission, which was based on R. rickettsii-infected and -uninfected A. aureolatum ticks feeding simultaneously on guinea pigs immune to R. rickettsii. Our acquisition and transmission infestations demonstrated that horizontal transmission of R. rickettsii by co-feeding ticks on immune hosts with no systemic infection did not occur when uninfected larvae fed distantly from infected nymphs but did occur in a few cases when uninfected larvae fed side-by-side with infected nymphs, suggesting that they shared the same feeding site. The co-feeding nonsystemic transmission type might have no epidemiologic importance for Rocky Mountain spotted fever.
Asunto(s)
Vectores Arácnidos/microbiología , Ixodidae/microbiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Animales , Brasil , Susceptibilidad a Enfermedades , Femenino , Cobayas , Humanos , Larva , Ninfa , Fiebre Maculosa de las Montañas Rocosas/microbiologíaRESUMEN
The clinical and pathologic characterisation of two fatal cases of tick-borne rickettsiosis in rural (El Valle) and urban (City of Panama) Panama are described. Clinical and autopsy findings were non-specific, but the molecular analysis was used to identify Rickettsia rickettsii in both cases. No ticks were collected in El Valle, while in the urban case, R. rickettsii was detected in Rhipicephalus sanguineus s.l., representing the first molecular finding in this tick in Panama and Central America.
Asunto(s)
Rhipicephalus sanguineus/microbiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/diagnóstico , Adolescente , Animales , Niño , Resultado Fatal , Humanos , Masculino , Panamá , Fiebre Maculosa de las Montañas Rocosas/microbiologíaRESUMEN
Rickettsia rickettsii is a tick-borne obligate intracellular bacterium that causes Rocky Mountain Spotted Fever (RMSF). In Brazil, two species of ticks in the genus Amblyomma, A. sculptum and A. aureolatum, are incriminated as vectors of this bacterium. Importantly, these two species present remarkable differences in susceptibility to R. rickettsii infection, where A. aureolatum is more susceptible than A. sculptum. In the current study, A. aureolatum and A. sculptum ticks were fed on suitable hosts previously inoculated with R. rickettsii, mimicking a natural infection. As control, ticks were fed on non-infected animals. Both midgut and salivary glands of all positively infected ticks were colonized by R. rickettsii. We did not observe ticks with infection restricted to midgut, suggesting that important factors for controlling rickettsial colonization were produced in this organ. In order to identify such factors, the total RNA extracted from the midgut (MG) was submitted to next generation RNA sequencing (RNA-seq). The majority of the coding sequences (CDSs) of A. sculptum differentially expressed by infection were upregulated, whereas most of modulated CDSs of A. aureolatum were downregulated. The functional categories that comprise upregulated CDSs of A. sculptum, for instance, metabolism, signal transduction, protein modification, extracellular matrix, and immunity also include CDSs of A. aureolatum that were downregulated by infection. This is the first study that reports the effects of an experimental infection with the highly virulent R. rickettsii on the gene expression of two natural tick vectors. The distinct transcriptional profiles of MG of A. sculptum and A. aureolatum upon infection stimulus strongly suggest that molecular factors in this organ are responsible for delineating the susceptibility to R. rickettsii. Functional studies to determine the role played by proteins encoded by differentially expressed CDSs in the acquisition of R. rickettsii are warranted and may be considered as targets for the development of strategies to control the tick-borne pathogens as well as to control the tick vectors.
Asunto(s)
Susceptibilidad a Enfermedades , Interacciones Huésped-Patógeno/genética , Ixodidae/genética , Ixodidae/microbiología , Rickettsia rickettsii/fisiología , Rickettsia rickettsii/patogenicidad , Garrapatas/genética , Garrapatas/microbiología , Animales , Brasil , Vectores de Enfermedades , Conducta Alimentaria , Femenino , Tracto Gastrointestinal/microbiología , Regulación de la Expresión Génica , Ixodidae/inmunología , Ixodidae/metabolismo , Sistemas de Lectura Abierta/genética , Rickettsia rickettsii/crecimiento & desarrollo , Fiebre Maculosa de las Montañas Rocosas/microbiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Glándulas Salivales/microbiología , Análisis de Secuencia de ARN , Transducción de Señal , Garrapatas/metabolismo , Transcriptoma/genética , VirulenciaRESUMEN
The aim of this work was to test the vectorial competence of Amblyomma tonelliae (Ixodida: Ixodidae) to transmit Rickettsia rickettsii (Rickettsiales: Rickettsiaceae), the agent of Rocky Mountain spotted fever (RMSF). All parasitic stages of A. tonelliae were exposed to R. rickettsii by allowing each stage to feed on hosts inoculated with this pathogen. Thereafter, ticks were fed on uninfected hosts. All stages of A. tonelliae were able to acquire the R. rickettsii infection and maintain it by transstadial and transovarial transmission. When infected ticks fed on uninfected hosts, the hosts developed rickettsiosis disease. This study demonstrates the vectorial competence of A. tonelliae to transmit R. rickettsii. These results have epidemiological relevance because A. tonelliae is one of the tick species most likely to infest humans in Argentina, including in areas in which RMSF has been reported.
Asunto(s)
Vectores Arácnidos/fisiología , Ixodidae/fisiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Animales , Vectores Arácnidos/crecimiento & desarrollo , Vectores Arácnidos/microbiología , Argentina , Conducta Alimentaria , Femenino , Ixodidae/crecimiento & desarrollo , Ixodidae/microbiología , Larva/crecimiento & desarrollo , Larva/microbiología , Larva/fisiología , Ninfa/crecimiento & desarrollo , Ninfa/microbiología , Ninfa/fisiologíaRESUMEN
BACKGROUND: Rickettsia rickettsii, the etiological agent of Rocky Mountain spotted fever, is transmitted to humans by ticks. During tick feeding, R. rickettsii is exposed to both temperature elevation and components of the blood meal, which have previously been associated with the reactivation of its virulence. These environmental stimuli were also reported to modulate virulence genes of R. rickettsii infecting a set of organs of adult females of its natural vector, Amblyomma aureolatum. METHODS: In this study, we determined the effects of a temperature upshift, blood-feeding, and both stimuli simultaneously on the expression of 85 selected genes of R. rickettsii infecting either the midgut (MG) or salivary glands (SG) of male and female A. aureolatum by microfluidic high-throughput RT-qPCR. These two organs are key for acquisition of this bacterium by the tick and transmission to the vertebrate host, respectively. RESULTS: Data showed that these environmental stimuli exert distinct effects on rickettsial transcription depending on the colonized organ and gender of the vector. Temperature upshift induced the majority of differentially expressed genes of R. rickettsii in tick SG, including tRNA synthetases encoding genes. On the contrary, blood-feeding downregulated most of differentially expressed genes in both organs, but induced type IV secretion system components and OmpB in tick MG. The combined effects of both stimuli resulted in a merged gene expression profile representing features of each stimulus analyzed independently, but was more similar to the profile induced by blood-feeding. CONCLUSION: The upregulation of the majority of differentially expressed genes in tick SG by temperature upshift suggests that this stimulus is important to prepare R. rickettsii for transmission to the vertebrate host. Blood-feeding, on the other hand, induced important virulence genes in the tick MG, which might be associated with colonization of the tick and transmission to the vertebrate host. The role of the proteins identified in this study must be addressed and might help to define future targets to block tick infection, thereby preventing RMSF. To our knowledge, this is the first transcriptional tissue-specific study of a virulent strain of R. rickettsii infecting a natural tick vector.
Asunto(s)
Tracto Gastrointestinal/microbiología , Regulación Bacteriana de la Expresión Génica/fisiología , Ixodidae/microbiología , Rickettsia rickettsii/fisiología , Temperatura , Animales , Conducta Alimentaria , Femenino , Interacciones Huésped-Patógeno , Ixodidae/fisiología , Masculino , Rickettsia rickettsii/genética , Virulencia/genéticaRESUMEN
The hard tick Amblyomma aureolatum (Pallas) is a vector of the bacterium Rickettsia rickettsii, the etiologic agent of Brazilian spotted fever (BSF) in parts of Brazil. Despite its wide distribution in southeastern South America and its public health importance, there is no information about genetic variation of this species that might help to understand the epidemiology of BSF. Using data from eight microsatellite markers and ticks from six localities, we used a population genetics approach to test the hypothesis that tick populations from areas with the presence of R. rickettsii are genetically different from ticks from areas without R. rickettsii Contrary to expectations, we found low genetic structure between studied regions. Thus, the presence of R. rickettsii in the specific area is more likely correlated with ecological and the environmental conditions or due to unknown gene coding regions of A. aureolatum genome that would be related to R. rickettsii infection resistance.
Asunto(s)
Variación Genética , Ixodidae/fisiología , Repeticiones de Microsatélite , Animales , Brasil , Ixodidae/genética , Ixodidae/microbiología , Dinámica Poblacional , Rickettsia rickettsii/fisiologíaRESUMEN
Amblyomma aureolatum (Pallas) and Amblyomma ovale Koch are common ectoparasites of domestic dogs in São Paulo state, southeastern Brazil, where they are vectors of distinct spotted fever group rickettsioses, one caused by Rickettsia rickettsii (transmitted by A. aureolatum), and the other caused by Rickettsia sp. strain Atlantic rainforest (transmitted by A. ovale). For the present study, we performed an altitudinal assessment of all 1992-2012 records of A. aureolatum and A. ovale retrieved from a tick collection. The municipalities with A. ovale records presented significantly (P < 0.05) lower altitude than the ones with A. aureolatum records; the higher the altitude, the lower the chances for the occurrence of A. ovale and the greater the likelihood for the occurrence of A. aureolatum. Regarding A. aureolatum, the chances of finding it in municipalities between 101 and 700 m are nine times higher than in municipalities at ≤ 100 m, or 31.5 times higher in municipalities above 700 m, when compared with municipalities at ≤ 100 m. The reverse was observed for A. ovale, which had its odds ratio diminishing at higher altitudes. These findings have a major role to public health, as A. aureolatum is associated with the transmission of a highly lethal spotted fever (caused by R. rickettsii), whereas A. ovale is associated with the transmission of a milder spotted fever (caused by Rickettsia sp. strain Atlantic rainforest, a R. parkeri-like agent), both in the state of São Paulo.
Asunto(s)
Distribución Animal , Vectores Arácnidos/fisiología , Ixodidae/fisiología , Rickettsia/fisiología , Altitud , Animales , Brasil , Ecosistema , Infecciones por Rickettsia/microbiología , Infecciones por Rickettsia/transmisión , Rickettsia rickettsii/fisiología , Especificidad de la EspecieRESUMEN
Amblyomma dubitatum engorged females, naturally infected by Rickettsia bellii, were used to establish a laboratory colony. Larvae, nymphs, and adults were exposed to two strains of Rickettsia rickettsii by feeding on needle-inoculated guinea pigs, and thereafter reared on uninfected guinea pigs. After acquisition feeding, engorged larvae and nymphs molted to nymphs and adults, respectively, which were shown to be infected (confirming transstadial perpetuation), and were able to transmit both strains of R. rickettsii to uninfected animals, as demonstrated by clinical, serological, and molecular analyses. However, the larval, nymphal, and adult stages of A. dubitatum showed to be only partially susceptible to R. rickettsii infection, since in all cases, only part of the ticks became infected by this agent, after being exposed to rickettsemic animals. While transovarial transmission of R. rickettsii was inefficient in the A. dubitatum engorged females of the present study, 100% of these females passed R. bellii transovarially. Because it has been reported that a primary infection by a Rickettsia species would preclude transovarial transmission of a second Rickettsia species, it is likely that the ineffectiveness of A. dubitatum to perpetuate R. rickettsii by transovarial transmission was related to its primary infection by R. bellii; however, it could also be related to unknown factors inherent to A. dubitatum. The relevance of A. dubitatum as a natural vector of R. rickettsii to humans or animals is discussed.
Asunto(s)
Vectores Arácnidos/microbiología , Ixodidae/microbiología , Rickettsia rickettsii/fisiología , Rickettsia/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Animales , Vectores Arácnidos/fisiología , Femenino , Cobayas , Humanos , Ixodidae/fisiología , Larva , Muda , Ninfa , Fiebre Maculosa de las Montañas Rocosas/microbiologíaRESUMEN
Rickettsia rickettsii is an obligate intracellular tick-borne bacterium that causes Rocky Mountain Spotted Fever (RMSF), the most lethal spotted fever rickettsiosis. When an infected starving tick begins blood feeding from a vertebrate host, R. rickettsii is exposed to a temperature elevation and to components in the blood meal. These two environmental stimuli have been previously associated with the reactivation of rickettsial virulence in ticks, but the factors responsible for this phenotype conversion have not been completely elucidated. Using customized oligonucleotide microarrays and high-throughput microfluidic qRT-PCR, we analyzed the effects of a 10°C temperature elevation and of a blood meal on the transcriptional profile of R. rickettsii infecting the tick Amblyomma aureolatum. This is the first study of the transcriptome of a bacterium in the genus Rickettsia infecting a natural tick vector. Although both stimuli significantly increased bacterial load, blood feeding had a greater effect, modulating five-fold more genes than the temperature upshift. Certain components of the Type IV Secretion System (T4SS) were up-regulated by blood feeding. This suggests that this important bacterial transport system may be utilized to secrete effectors during the tick vector's blood meal. Blood feeding also up-regulated the expression of antioxidant enzymes, which might correspond to an attempt by R. rickettsii to protect itself against the deleterious effects of free radicals produced by fed ticks. The modulated genes identified in this study, including those encoding hypothetical proteins, require further functional analysis and may have potential as future targets for vaccine development.
Asunto(s)
Conducta Alimentaria/fisiología , Perfilación de la Expresión Génica , Insectos Vectores/microbiología , Rickettsia rickettsii/genética , Rickettsia rickettsii/fisiología , Temperatura , Garrapatas/microbiología , Animales , Sistemas de Secreción Bacterianos/genética , Simulación por Computador , Femenino , Genes Bacterianos/genética , Cobayas , Microfluídica , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reproducibilidad de los Resultados , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Infecciones por Rickettsia/genética , Infecciones por Rickettsia/microbiologíaRESUMEN
The tick-borne bacterium Rickettsia rickettsii is the aetiological agent of Brazilian spotted fever (BSF). The present study evaluated tick infestations on wild and domestic animals, and the rickettsial infection in these animals and their ticks in 7 forest areas adjacent to human communities in the São Paulo Metropolitan Area (SPMA). The results were compared to ecological traits of each sampled area. Two main tick species, Amblyomma aureolatum and Rhipicephalus sanguineus, were collected from dogs. The major ticks found on small mammals and birds were Ixodes loricatus and Amblyomma longirostre, respectively. Both anti-R. rickettsii antibodies and R. rickettsii-infected ticks were detected on dogs from only 2 areas in the southern part of the SPMA, which were considered to be endemic for BSF; the remaining 5 areas were considered to be non-endemic. Ecologically, the BSF-endemic areas clearly differed from the non-endemic areas by the presence of significantly more degraded forest patches in the former. The present results corroborate historical observations that have indicated that all human cases of BSF in the SPMA were contracted in the southern part of this metropolitan area. However, not all forest patches in the southern part of the SPMA were shown to be associated with BSF endemism.
Asunto(s)
Enfermedades de las Aves/epidemiología , Enfermedades de los Perros/epidemiología , Fiebre Maculosa de las Montañas Rocosas/epidemiología , Árboles , Animales , Animales Salvajes , Anticuerpos Antibacterianos/sangre , Biodiversidad , Aves , Brasil/epidemiología , Enfermedades de los Perros/microbiología , Perros , Ixodidae/microbiología , Mamíferos , Datos de Secuencia Molecular , Enfermedades Parasitarias en Animales/epidemiología , Enfermedades Parasitarias en Animales/microbiología , Prevalencia , ARN Ribosómico 16S , Rhipicephalus sanguineus/microbiología , Rickettsia rickettsii/genética , Rickettsia rickettsii/aislamiento & purificación , Rickettsia rickettsii/fisiología , Estudios Seroepidemiológicos , Infestaciones por Garrapatas/epidemiología , Infestaciones por Garrapatas/microbiologíaRESUMEN
In the laboratory, Amblyomma cajennense (Acari: Ixodidae) (Fabricius) larvae, nymphs and adults were exposed to Rickettsia rickettsii by feeding on needle-inoculated animals, and thereafter reared on uninfected guinea pigs or rabbits. Regardless of the tick stage that acquired the infection, subsequent tick stages were shown to be infected (confirming transstadial and transovarial transmissions) and were able to transmit R. rickettsii to uninfected animals, as demonstrated by serological and molecular analyses. However, the larval, nymphal and adult stages of A. cajennense were shown to be partially refractory to R. rickettsii infection, as in all cases, only part of the ticks became infected by this agent, after being exposed to rickettsemic animals. In addition, less than 50% of the infected engorged females transmitted rickettsiae transovarially, and when they did so, only part of the offspring became infected, indicating that vertical transmission alone is not enough to maintain R. rickettsii in A. cajennense for multiple generations. Finally, the R. rickettsii-infected tick groups had lower reproductive performance than the uninfected control group. Our results indicate that A. cajennense have a low efficiency to maintain R. rickettsii for successive generations, as R. rickettsii-infection rates should decline drastically throughout the successive tick generations.
Asunto(s)
Vectores Arácnidos/microbiología , Insectos Vectores/microbiología , Ixodidae/microbiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Animales , Vectores Arácnidos/fisiología , Brasil , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Cobayas , Interacciones Huésped-Parásitos , Insectos Vectores/fisiología , Ixodidae/crecimiento & desarrollo , Larva/crecimiento & desarrollo , Larva/microbiología , Masculino , Ninfa/crecimiento & desarrollo , Ninfa/microbiología , Reacción en Cadena de la Polimerasa , Conejos , Fiebre Maculosa de las Montañas Rocosas/microbiologíaRESUMEN
We experimentally infected Amblyomma aureolatum ticks with the bacterium Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (RMSF). These ticks are a vector for RMSF in Brazil. R. rickettsii was efficiently conserved by both transstadial maintenance and vertical (transovarial) transmission to 100% of the ticks through 4 laboratory generations. However, lower reproductive performance and survival of infected females was attributed to R. rickettsii infection. Therefore, because of the high susceptibility of A. aureolatum ticks to R. rickettsii infection, the deleterious effect that the bacterium causes in these ticks may contribute to the low infection rates (<1%) usually reported among field populations of A. aureolatum ticks in RMSF-endemic areas of Brazil. Because the number of infected ticks would gradually decrease after each generation, it seems unlikely that A. aureolatum ticks could sustain R. rickettsii infection over multiple successive generations solely by vertical transmission.
Asunto(s)
Rickettsia rickettsii/fisiología , Garrapatas/microbiología , Animales , Perros , Femenino , Cobayas , Masculino , Conejos , Fiebre Maculosa de las Montañas Rocosas/microbiología , Fiebre Maculosa de las Montañas Rocosas/transmisiónRESUMEN
We evaluated if Rickettsia rickettsii-experimentally infected dogs could serve as amplifier hosts for hipicephalus sanguineus ticks. In addition, we checked if Rh. sanguineus ticks that acquired Ri. rickettsii from dogs could transmit the bacterium to susceptible hosts (vector competence), and if these ticks could maintain the bacterium by transstadial and transovarial transmissions. Uninfected larvae, nymphs, and adults of Rh. sanguineus were allowed to feed upon three groups of dogs: groups 1 (G1) and 2 (G2) composed of Ri. rickettsii-infected dogs, infected intraperitoneally and via tick bites, respectively, and group 3 composed of uninfected dogs. After larval and nymphal feeding on rickettsemic dogs, 7.1-15.2% and 35.8-37.9% of the molted nymphs and adults, respectively, were shown by polymerase chain reaction (PCR) to be infected by Ri. rickettsii, confirming that both G1 and G2 dogs were efficient sources of rickettsial infection (amplifier host), resulting in transstadial transmission of the agent. These infected nymphs and adults successfully transmitted Ri. rickettsii to guinea pigs, confirming vector competence after acquisition of the infection from rickettsemic dogs. Transovarial transmission of Ri. rickettsii was observed in engorged females that had been infected as nymphs by feeding on both G1 and G2 dogs, but not in engorged females that acquired the infection during adult feeding on these same dogs. In the first case, filial infection rates were generally <50%. No tick exposed to G3 dogs was infected by rickettsiae in this study. No substantial mortality difference was observed between Ri. rickettsii-infected tick groups (G1 and G2) and uninfected tick group (G3). Our results indicate that dogs can be amplifier hosts of Ri. rickettsii for Rh. sanguineus, although only a minority of immature ticks (<45%) should become infected. It appears that Rh. sanguineus, in the absence of horizontal transmission, would not maintain Ri. rickettsii through successive generations, possibly because of low filial infection rates.
Asunto(s)
Enfermedades de los Perros/transmisión , Rhipicephalus sanguineus/microbiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Animales , Vectores Arácnidos/microbiología , Brasil , Enfermedades de los Perros/microbiología , Perros , Femenino , Cobayas , Interacciones Huésped-Parásitos , Larva/microbiología , Ninfa/microbiología , Reacción en Cadena de la Polimerasa , Fiebre Maculosa de las Montañas Rocosas/microbiología , Fiebre Maculosa de las Montañas Rocosas/veterinariaRESUMEN
The present study evaluated the infection of capybaras (Hydrochoerus hydrochaeris) by Rickettsia rickettsii and their role as amplifier hosts for horizontal transmission of R. rickettsii to Amblyomma cajennense ticks. Two groups of two capybaras each were evaluated: on day 0, group 1 (G1) was infested by R. rickettsii-infected ticks, and group 2 (G2) was inoculated intraperitoneally with R. rickettsii. Two additional groups were control groups, not exposed to R. rickettsii, being CG1 group the control of G1, and CG2 group the control of G2. Capybara rectal temperature was measured daily. Blood samples were collected every 3 days during 30 days, and used to (i) inoculate guinea pigs intraperitoneally; (ii) DNA extraction followed by real-time PCR targeting the rickettsial gene gltA; (iii) hematology; (iv) detection of R. rickettsii-reactive antibodies by indirect immunofluorescence assay (IFA). Blood was also collected from G1 capybaras every approximately 10-30 days till the 146th day, to be tested by serology. Capybaras were infested by uninfected A. cajennense nymphs from the 3rd to the 18th day. Engorged nymphs were collected, allowed to molt to adults in an incubator. Thereafter, the subsequent flat ticks were tested by PCR. All G1 and G2 capybaras became infected by R. rickettsii, as demonstrated by guinea pig inoculation and seroconversion, but they showed no fever. Rickettsemia was continually detected from the 6th (G2 capybaras) or 9th (G1 capybaras) to the 18th day post inoculation or infestation with R. rickettsii-infected ticks. A total of 20-25% and 30-35% of the flat ticks previously fed on G1 and G2 capybaras, respectively, became infected by R. rickettsii. The study demonstrated that R. rickettsii was capable to infect capybaras without causing clinical illness, inducing rickettsemia capable to cause infection in guinea pigs and ticks. Our results indicate that capybaras act as amplifier host of R. rickettsii for A. cajennense ticks in Brazil.
Asunto(s)
Ixodidae/microbiología , Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/veterinaria , Roedores/microbiología , Animales , Cobayas , Ninfa/microbiología , Fiebre Maculosa de las Montañas Rocosas/microbiología , Fiebre Maculosa de las Montañas Rocosas/transmisiónRESUMEN
The present study evaluated the infection of opossums (Didelphis aurita) by Rickettsia rickettsii and their role as amplifier hosts for horizontal transmission of R. rickettsii to Amblyomma cajennense ticks. Three groups of opossums were evaluated: on day 0, group 1 (G1) was inoculated intraperitoneally with R. rickettsii; group 2 (G2) was infested by R. rickettsii-infected ticks; and group 3 (G3) was the uninfected control group. Opossum rectal temperature was measured daily. Blood samples were collected every 2 to 4 days during 30 days, and used to (1) inoculate guinea pigs intraperitoneally; (2) extract DNA followed by real-time polymerase chain reaction (PCR) targeting the rickettsial gene gltA; (3) study hematology; (4) detect R. rickettsii-reactive antibodies by indirect immunofluorescence assay (IFA). Blood was also collected every 10 days from days 30 to 180, to be tested by serology. Opossums were infested by uninfected A. cajennense larvae and nymphs from days 3 to 15. Engorged ticks were collected and allowed to molt in an incubator. Thereafter, the subsequent flat ticks were allowed to feed on uninfected rabbits, which were tested for seroconversion by IFA. Samples of flat ticks were also tested by real-time PCR. All G1 and G2 opossums became infected by R. rickettsii, as demonstrated by realtime PCR or/and guinea pig inoculation, but they showed no clinical abnormality. Rickettsemia was first detected at days 2 to 8, lasting intermittently till days 1 to 30. Approximately 18% and 5% of the flat ticks previously fed on G1 and G2 opossums, respectively, became infected by R. rickettsii, but only the rabbits infested with G1-derived ticks seroconverted. The study demonstrated that R. rickettsii was capable of infecting opossums without causing illness and developing rickettsemia capable of causing infection in guinea pigs and ticks, although the infection rate in ticks was low.