RESUMO

Adult Amblyomma sculptum and Amblyomma aureolatum ticks are partially refractory to Rickettsia rickettsii when fed on infected hosts, hindering the functional characterization of potentially protective targets in the bacterial acquisition. In the current study, we used the anal pore route to infect adult A. sculptum and A. aureolatum ticks with R. rickettsii and to assess the effects of the knockdown of microplusin in infection control. The anal pore route was efficient to infect both species, resulting in a prevalence of around 100% of infected ticks. Higher loads of R. rickettsii were detected in microplusin-silenced A. aureolatum in relation to the control, as previously obtained when microplusin-silenced ticks were fed on R. rickettsii-infected rabbits. This is the first report showing R. rickettsii infection through the anal pore in Amblyomma ticks, highlighting this route as a powerful tool to assess the role played by additional targets in the control of pathogens.

Assuntos

RESUMO

Rocky Mountain spotted fever (RMSF), caused by the bacterium Rickettsia rickettsii, is a re-emerging tick-borne zoonosis in North America, with hundreds of human fatalities in multiple outbreaks in northern Mexico and the southwestern US in the past few decades. Free-roaming dogs are key because they are reservoirs for the pathogen and the main hosts of the brown dog tick (Rhipicephalus sanguineus), which vectors RMSF in this region. Because coyotes (Canis latrans) can be infected with R. rickettsii and infested with Rh. sanguineus, we hypothesized that space sharing among dogs and coyotes could enhance disease risks. In summer 2021, we captured and sampled 11 coyotes at two sites in Baja California, Mexico, near population centers with human cases of RMSF, and fitted seven individuals with GPS logging collars. We also tested tissue samples, sera, and ectoparasites for DNA of R. rickettsii with PCR and used serology to detect antibodies to R. rickettsii. Finally, we deployed an array of cameras to document dog-coyote interactions. Mean home range size was 40.37 km2. Both GPS and camera data showed considerable home range overlap both between individual coyotes and between coyotes and dogs. Coyotes were active in areas where dogs occur including the domestic interface surrounding human settlements. Although none of our samples were positive for R. rickettsii on PCR, 72.7% (8/11) of the samples were seropositive with titers ≥64. Our data confirm shared space use and risk of shared parasites and disease between coyotes and dogs.

Assuntos

RESUMO

We found serologic evidence of spotted fever group Rickettsia in humans and dogs and typhus group Rickettsia in dogs in Reynosa, Mexico. Our investigation revealed serologic samples reactive to spotted fever group Rickettsia in 5 community members, which highlights a potential rickettsial transmission scenario in this region.

Assuntos

RESUMO

Twelve tick species have been reported in El Salvador; however, information regarding ticks infesting domestic dogs is lacking, and pathogenic tick-borne Rickettsia species have never been reported in El Salvador. This work evaluated ticks infesting 230 dogs from ten municipalities in El Salvador from July 2019 to August 2020. A total of 1,264 ticks were collected and identified into five species: Rhipicephalus sanguineus sensu lato (s.l.), Rhipicephalus microplus, Amblyomma mixtum, Amblyomma ovale, and Amblyoma cf. parvum. The tick R. sanguineus s.l. was the most frequent species in all localities (81.3% of sampled dogs), followed by Amblyomma mixtum (13.0%), Amblyomma ovale (10.9%) and Amblyomma cf. parvum (10.4%). The overall mean intensity of tick infestation was 5.5 ticks/dog. The highest specific mean intensity value was for R. sanguineus s.l. (4.8 ticks/dog), varying from 1.6 to 2.7 ticks/dog for the three Amblyomma species. From a random sample of 288 tick specimens tested molecularly for the presence of rickettsial agents, three spotted fever group Rickettsia were detected: Rickettsia amblyommatis in 90% (36/40) A. mixtum, 46% (11/24) A. cf. parvum, 4% (7/186) R. sanguineus s.l., and 17% Amblyomma spp.; Rickettsia parkeri strain Atlantic rainforest in 4% (1/25) A. ovale; and an unnamed rickettsia agent, designated as 'Rickettsia sp. ES-A.cf.parvum', in 4% (1/24) A. cf. parvum. Our finding of R. parkeri strain Atlantic rainforest in A. ovale is highly relevant because this agent has been associated to spotted fever illness in other Latin American countries, where A. ovale is implicated as its main vector. These findings suggest that spotted fever cases caused by R. parkeri strain Atlantic rainforest could be occurring in El Salvador.

Assuntos

RESUMO

Ticks parasitizing 102 wild animals in the states of Mato Grosso and Goiás, Brazil were collected between 2015 and 2018. A total of 2338 ticks (865 males, 541 females, 823 nymphs, and 109 larvae) belonging to four genera (Amblyomma, Dermacentor, Haemaphysalis, and Rhipicephalus) and at least 21 species were identified. DNA extraction and a molecular survey for rickettsial agents were performed on 650 ticks. The results revealed parasitism by the following species: Rickettsia amblyommatis in Amblyomma cajennense s.s., A. cajennense s.l., Amblyomma coelebs, Amblyomma humerale, Amblyomma longirostre, Amblyomma nodosum, Amblyomma scalpturatum, Amblyomma sculptum, and Amblyomma romitii; Rickettsia parkeri in Amblyomma nodosum, Amblyomma ovale, Amblyomma scalpturatum, and Amblyomma triste; Rickettsia rhipicephali in Haemaphysalis juxtakochi; Rickettsia sp. in A. cajennense s.s., A. nodosum, and A. sculptum, and lastly, 'Candidatus Rickettsia andeanae' in Amblyomma parvum and Rhipicephalus microplus. This study expands the body of knowledge about tick parasitism among wild animals, including new data concerning tick-host associations, and provides information about the epidemiology of tick-borne pathogens in the Center-West region of Brazil.(AU)

Foram coletados carrapatos parasitando 102 animais silvestres nos estados de Mato Grosso e Goiás, Brasil, entre 2015 e 2018. Um total de 2338 carrapatos (865 machos, 541 fêmeas, 823 ninfas e 109 larvas) pertencentes a quatro gêneros (Amblyomma, Dermacentor, Haemaphysalis e Rhipicephalus) e pelo menos 21 espécies foram identificadas. A extração de DNA e pesquisa molecular para agentes riquétsiais foram realizadas em 650 carrapatos. Os resultados revelaram parasitismo pelas seguintes espécies: Rickettsia amblyommatis em Amblyomma cajennense s.s., A. cajennense s.l., Amblyomma coelebs, Amblyomma humerale, Amblyomma longirostre, Amblyomma nodosum, Amblyomma scalpturatum, Amblyomma sculptum e Amblyomma romitii; Rickettsia parkeri em Amblyomma nodosum, Amblyomma ovale, Amblyomma scalpturatum e Amblyomma triste; Rickettsia rhipicephali em Haemaphysalis juxtakochi; Rickettsia sp. em A. cajennense s.s., A. nodosum e A. sculptum e, por último, 'Candidatus Rickettsia andeanae' em Amblyomma parvum e Rhipicephalus microplus. Este estudo amplia o conhecimento sobre o parasitismo de carrapatos em animais silvestres, incluindo-se novos dados sobre associações carrapato-hospedeiro. Fornece, ainda, informações sobre a epidemiologia de patógenos transmitidos por carrapatos na região Centro-Oeste do Brasil.(AU)

Assuntos

RESUMO

We conducted enhanced acute febrile illness surveillance in an urban slum community in Salvador, Brazil. We found that rickettsial infection accounted for 3.5% of urgent care visits for acute fever. Our results suggest that rickettsiae might be an underrecognized, treatable cause of acute febrile illness in impoverished urban populations in Brazil.

Assuntos

RESUMO

The pivotal role of amphibians in food webs and their value as indicators of disequilibrium in ecosystem health have long been recognized by wildlife biologists. However, massive pathogen-induced declines in global amphibian populations reported during the last 30 years served to alert the scientific community that knowledge of amphibian disease ecology, including parasitic and vector-borne conditions, was and remains incipient. Herein, we report the detection of a Rickettsia bacterium infecting larvae of the argasid tick Ornithodoros faccinii, collected from the toad Rhinella ornata, in Southeastern Brazil. Fragments of the genes 16S rDNA, gltA, htrA, sca1, sca4, and ompB were amplified by polymerase chain reaction (PCR), but the sequence encoding the ompA antigen was not detected. Nucleotide sequencing and multi-locus (gltA, htrA, sca1, and sca4) phylogenetic analyses characterized the bacterium, designated Rickettsia sp. strain Itinguçú, as a novel member of the spotted fever group (SFG) of the Rickettsia, closely related to the Rickettsia massiliae and to a lesser extent the Rickettsia helvetica subgroups. The apparent absence of the ompA protein together with limited levels of nucleotide (90.5 %) and amino acid (82-83 %) sequence identity, relative to the ompB gene of other species in the R. massiliae subgroup, were unusual features that may reflect adaptation to selective pressures exerted by the tick and/or amphibian immune systems. The ompB sequence was exploited to develop a low-cost method for differential identification of Rickettsia sp. strain Itinguçú, based on restriction fragment length polymorphism analysis of amplicons (PCR-RFLP). The characterization of this novel bacterium provided an unprecedented record of infection by an SFG Rickettsia in a member of the family Argasidae infesting a cold-blooded animal and raised the number of tick-associated Rickettsia reported in Brazil to sixteen. Moreover, it highlighted the value of and the requirement for continued and extended surveillance of wildlife as potential sources of emerging tick-borne pathogens.

Assuntos

RESUMO

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.

Assuntos

RESUMO

The present study was carried out to identify Rickettsia species with zoonotic potential in ticks collected from dogs in a rural area in Tabasco, Mexico. In total 197 Amblyomma maculatum ticks were collected from 40 domestic dogs. The collected specimens were pooled and subjected to DNA extraction. A fragment (380 bp) of citrate synthase gene (gltA) was amplified by polymerase chain reaction (PCR) using universal primers for Rickettsia. A second PCR was later performed to amplify a fragment (420 bp) of the outer membrane protein B gene (ompB). The PCR products were purified, sequenced and compared using the basic local alignment search tool (BLAST). Twenty out of 40 (50%) tick pools assayed were positive for rickettsial DNA using both primer pairs. The consensus sequence obtained from the ompB gene fragments showed 99.5-100% of identity with strains of Rickettsia parkeri. This study provides the first molecular evidence of the presence of R. parkeri in A. maculatum ticks infesting domestic dogs from southeastern Mexico. Close contact between dogs and humans should lead to consider the infection caused by this species of Rickettsia among the differential diagnoses for people of Tabasco, Mexico, who show acute febrile syndrome associated to inoculation eschar and have a clinical history of tick exposure.

Assuntos

RESUMO

Introduction of exotic tick vectors of bacteria, protozoa, viruses, and filarial parasites into the United States has accelerated in recent years, primarily because of globalization, increased frequency of travel, and a rise in legal and illegal animal trades. We herein report introduction of a live specimen of Amblyomma oblongoguttatum on a human into the United States from Central America, and we review 4 previous similar incidents. This tick species occurs widely in the neotropics, from western and southern Mexico, southwards through Central America, to the northern half of South America. It is a potential vector of bacterial agents of spotted fever group rickettsioses, raising concern that if A. oblongoguttatum ticks become established in this country, they might also be able to carry pathogens of human and veterinary concern. Given the potential for exotic ticks as vectors of numerous pathogens, proper surveillance, interception, and identification of these ticks are vital to protecting human and veterinary health. Rigorous governmental inspections of imported livestock and pet animals at ports of entry and educating human travelers and medical practitioners about the risks should be part of an overall national tick program.

Assuntos

RESUMO

Q fever and rickettsial diseases occur throughout the world and appear to be emergent zoonoses in Chile. The diagnosis of these diseases is currently uncommon in Chile, as their clinical presentations are non-specific and appropriate diagnostic laboratory assays are of limited availability. During a recent outbreak of undiagnosed human atypical pneumonia, we serologically investigated a series of 357 cases from three regions of southern Chile. The aim was to identify those caused by Coxiella burnetii and/or Rickettsia spp. Serological analysis was performed by ELISA and an immunofluorescence assay (IFA) for acute and convalescence sera of patients. Our results, including data from two international reference laboratories, demonstrate that 71 (20%) of the cases were Q fever, and 44 (15%) were a likely rickettsial infection, although the rickettsial species could not be confirmed by serology. This study is the first report of endemic Q fever and rickettsial disease affecting humans in Chile.

RESUMO

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.

Assuntos

RESUMO

The bacterium Rickettsia bellii has been detected in 25 species of ticks in the American continents, but its pathogenic potential is considered as undetermined. A possible role for this species in the phenomenon of transovarial exclusion of pathogenic members of the spotted fever group (SFG) of Rickettsia has been suggested and co-infections with pathogenic species have been reported infrequently in both North and South America. Traditional methods for the molecular detection of rickettsial agents in ticks focus largely on the identification of sequences found in SFG Rickettsia, an approach that may overlook the presence of co-infections with R. bellii. Two novel, species-specific polymerase chain reaction (PCR) assays, targeting the genes encoding the surface cell antigen (Sca), autotransporter proteins sca9 and sca14, were developed and validated for the detection of R. bellii using 150 Amblyomma ticks collected from wild birds in Brazil. Co-infection of R. bellii infected ticks was evaluated using a novel PCR assay targeting the ompA sequence characteristic of SFG Rickettsia. Preliminary species-level identification was achieved by restriction fragment length polymorphism (RFLP) analysis and subsequently confirmed by sequencing of amplicons. Nine out of seventy-three Amblyomma longirostre and one of two Amblyomma calcaratum ticks were shown to be co-infected with R. bellii and Rickettsia amblyommatis, while two out of sixty-seven Amblyomma sp. haplotype Nazaré ticks were recorded as co-infected with R. bellii and the Rickettsia parkeri-like bacterium, strain ApPR. Interestingly, our data represent the first records of R. bellii in association with A. calcaratum and Amblyomma sp. haplotype Nazaré. The novel PCR-RFLP systems reported herein, provide an alternative, rapid and cost-efficient (relative to strategies based on sequencing or real-time PCR), approach to evaluate rickettsial co-infection of ticks, a potentially significant phenomenon that has most likely been underestimated to date.

Assuntos

RESUMO

Introduction of ticks into the United States that can carry disease-causing pathogens to humans, companion animals, and wildlife has accelerated in recent years, mostly due to globalization, frequency of travel, and a rise in legal and illegal animal trades. We hereby report for the first time introduction of a live fully engorged Amblyomma coelebs feeding on a human into the United States from Central America. Amblyomma coelebs is geographically distributed in the Neotropical region and reaches the southern states of Mexico. This species is capable of transmitting a number of pathogens of public health and veterinary importance including spotted fever group rickettsiae, raising concern that A. coelebs, if it became established in the United States, might also be able to carry these pathogens. Considering the risks of exotic ticks as vectors of numerous pathogens and their potential to establish new populations under conducive climatic and habitat conditions, rigorous inspection practices of imported livestock and pet animals at ports of entry are vital. It is also important for travelers and practitioners to develop a heightened awareness of the public health risks associated with the unintended importation of exotic ticks and the potential such parasites have for breaching United States biosecurity defenses.

Assuntos

RESUMO

Rickettsia parkeri sensu stricto (s.s.) is an emerging human pathogen in the Americas. Comprehension of the etiology of R. parkeri infections in South America is complicated by the existence of genetic variants (Atlantic rainforest, NOD and Parvitarsum) of this species that are associated with specific groups of Amblyomma ticks. The rickettsial bacterium strain ApPR was first reported in Amblyomma parkeri ticks in Southern Brazil in 2012 and was considered, based on sequencing of fragments of the gltA, htrA, ompA and ompB genes, to represent yet another genetic variant of R. parkeri. In the current work, a multi-locus phylogenetic analysis employing additional genes and intragenic regions was performed using DNA extracted from (a) larvae of A. parkeri and Amblyomma species haplotype Nazaré ticks collected from wild birds, (b) a nymph of Amblyomma sp. haplotype Nazaré recovered from a monkey (Callicebus nigrifons), representing the first report of that tick parasitizing a non-human primate and (c) from a cultured isolate of ApPR, isolated from colony-reared adults of Amblyomma geayi. Phylogenetic inference performed using Maximum-likelihood (ML), Maximum Parsimony (MP) and Bayesian (B) methods, consistently placed strain ApPR outside the New World R. parkeri complex and instead grouped it in proximity to the Old World species Rickettsia africae and Rickettsia sibirica. Estimates of evolutionary divergence provided additional support for the inferred phylogenetic relationship. Given the clear evolutionary distance between strain ApPR and R. parkeri we propose the recognition of "Candidatus Rickettsia paranaensis".

Assuntos

RESUMO

The salivary glands (SG) of ixodid ticks play a pivotal role in blood feeding, producing both the cement and the saliva. The cement is an adhesive substance that helps the attachment of the tick to the host skin, while the saliva contains a rich mixture of antihemostatic, anti-inflammatory, and immunomodulatory substances that allow ticks to properly acquire the blood meal. The tick saliva is also a vehicle used by several pathogens to be transmitted to the vertebrate host, including various bacterial species from the genus Rickettsia. Rickettsia rickettsii is a tick-borne obligate intracellular bacterium that causes the severe Rocky Mountain spotted fever. In Brazil, the dog yellow tick Amblyomma aureolatum is a vector of R. rickettsii. In the current study, the effects of an experimental infection with R. rickettsii on the global gene expression profile of A. aureolatum SG was determined by next-generation RNA sequencing. A total of 260 coding sequences (CDSs) were modulated by infection, among which 161 were upregulated and 99 were downregulated. Regarding CDSs in the immunity category, we highlight one sequence encoding one microplusin-like antimicrobial peptide (AMP) (Ambaur-69859). AMPs are important effectors of the arthropod immune system, which lack the adaptive response of the immune system of vertebrates. The expression of microplusin was confirmed to be significantly upregulated in the SG as well as in the midgut (MG) of infected A. aureolatum by a quantitative polymerase chain reaction preceded by reverse transcription. The knockdown of the microplusin expression by RNA interference caused a significant increase in the prevalence of infected ticks in relation to the control. In addition, a higher rickettsial load of one order of magnitude was recorded in both the MG and SG of ticks that received microplusin-specific dsRNA. No effect of microplusin knockdown was observed on the R. rickettsii transmission to rabbits. Moreover, no significant differences in tick engorgement and oviposition were recorded in ticks that received dsMicroplusin, demonstrating that microplusin knockdown has no effect on tick fitness. Further studies must be performed to determine the mechanism of action of this AMP against R. rickettsii.

RESUMO

We report Rickettsia parkeri and Candidatus Rickettsia andeanae in ticks of the Amblyomma maculatum group collected from dogs in Sonora, Mexico. Molecular characterization of these bacteria was accomplished by DNA amplification and sequence analysis of portions of the rickettsial genes gltA, htrA, ompA, and ompB.

Assuntos

RESUMO

Spotted fever (SF) is a tick-borne disease associated with Rickettsia spp.. In the Pampa biome, Southern Brazil, cases of SF seem to be strongly linked with the practice of hunting wild animals. An investigation of rickettsiae in tick species found on wild animals could provide more information regarding the rickettsiosis enzootic cycle. The aim of this study is to describe the results of a molecular survey of Rickettsia spp. in the Neotropical deer tick, Haemaphysalis juxtakochi Cooley, 1946 (Acari: Ixodidae), from the Brazilian Pampa. Ticks were obtained from 14 road-killed gray brocket deer, Mazama gouazoubira (Artiodactyla: Cervidae), found in nine different municipalities of Rio Grande do Sul state, Southern Brazil. Ticks were processed individually to obtain genomic DNA, and then Rickettsia spp. was investigated using a set of PCR reactions that amplified the rickettsial fragments of the gltA, ompA, and htrA genes. Of the 24 tick samples tested, DNA of Rickettsia parkeri sensu stricto (s.s.) was found in 11 H. juxtakochi specimens collected in two different areas of the Brazilian Pampa. This is the first report of R. parkeri s.s. (the main agent associated with SF in the Uruguayan, Argentinian, and Brazilian Pampa) in H. juxtakochi ticks. These findings indicate that R. parkeri s.s. may be much more common and widely distributed in the Pampa biome than previously assumed. Moreover, H. juxtakochi ticks and gray brocket deer could participate in the potential spillover of R. parkeri s.s. from endemic to non-endemic areas in the South American Pampa.

Assuntos

RESUMO

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.

Assuntos

RESUMO

We report molecular detection of Rickettsia africae in Amblyomma ovale ticks from Nicaragua and a novel rickettsial strain in an A. triste tick. Of 146 ticks from dogs, 16.4% were Rickettsia PCR positive. The presence of Rickettsia spp. in human-biting ticks in Nicaragua may pose a public health concern.

Assuntos