Ticks infesting terrestrial small mammals in a rural settlement in the Amazonas state, Brazil.

There is limited knowledge about tick diversity in the Amazon region. Here, we survey small terrestrial mammals for tick infestation at the Rio Pardo settlement, Amazonas State, Brazil. Sampling included rainy and dry seasons and four ecotones (primary forest, forest in regeneration, field crops and households). Each animal was inspected for ticks, which, if present, were placed in 70% alcohol and identified. Parasitological indexes were calculated and the presence/absence of ticks on hosts was tested for possible associations with independent variables (ecotone, host sex, host order, host family, host age and season). A total of 208 small mammals were captured, 47 individuals (10 species) in the primary forest, 124 (15 species) in the forest in regeneration, 11 (7 species) in the field crops, and 26 (4 species) in the households. A total of 14 small mammals were infested by ticks (overall prevalence: 6.7%; 95% CI: 3.72 - 11.04%), which consisted of 51 specimens that were identified into four species, as follows: Amblyomma humerale (32 nymphs); Ixodes luciae (6 females); Amblyomma coelebs (1 nymph); and Ornithodoros mimon (1 larva). In addition, 11 larvae were retained as Amblyomma spp. Only host order showed association (P = 0.002) with tick infestation, with marsupials 5.5 times more infested than rodents. Our record of O. mimon on D. marsupialis is the first on this host species, and the first record of a Argasidae tick in the Brazilian state of Amazonas. To the best of our knowledge, this is the first study that actively screened free-living terrestrial small mammals and provided data on prevalence, mean intensity and mean abundance of tick infestations in the Brazilian Amazonas state.

Isolation and molecular characterization of a novel relapsing fever group Borrelia from the white-eared opossum Didelphis albiventris in Brazil.

This study aimed to detect, isolate and to characterize by molecular methods a relapsing fever group (RFG) Borrelia in white-eared opossums (Didelphis albiventris) from Brazil. During 2015-2018, when opossums (Didelphis spp.) were captured in six municipalities of the state of São Paulo, Brazil, molecular analyses revealed the presence of a novel RFG Borrelia sp. in the blood of seven opossums (Didelphis albiventris), out of 142 sampled opossums (4.9% infection rate). All seven infected opossums were from a single location (Ribeirão Preto municipality). In a subsequent field study in Ribeirão Preto during 2021, two new opossums (D. albiventris) were captured, of which one contained borrelial DNA in its blood. Macerated tissues from this infected opossum were inoculated into laboratory animals (rodents and rabbits) and two big-eared opossums (Didelphis aurita), which had blood samples examined daily via dark-field microscopy. No spirochetes were visualized in the blood of the laboratory animals. Contrastingly, spirochetes were visualized in the blood of the two D. aurita opossums between 12 and 25 days after inoculation. Blood samples from these opossums were used for a multi-locus sequencing typing (MLST) based on six borrelial loci. Phylogenies inferred from MLST genes positioned the sequenced Borrelia genotype into the RFG borreliae clade basally to borreliae of the Asian-African group, forming a monophyletic group with another Brazilian isolate, "Candidatus B. caatinga". Based on this concatenated phylogenetic analysis, which supports that the new borrelial isolate corresponds to a putative new species, we propose the name "Candidatus Borrelia mimona".

One Health approach to Coxiella burnetii in Brazilian indigenous communities.

Indigenous health has posted complex challenges worldwide, particularly due to historical economic, territorial, social and environmental processes, which may lead to emergence and reemergence of pathogens. In addition to few Coxiella burnetii serosurveys in vulnerable populations, especially in developing tropical countries, no comprehensive One Health approach has focused on human-animal infection along with potential environmental determinants. Accordingly, this study aimed to assess the seroprevalence of anti-C. burnetii antibodies in indigenous populations and their dogs from 10 indigenous communities distributed in southern and southeastern Brazil, along with the correspondent healthcare professionals. In overall, 8/893 (0.90%; 95% CI 0.45-1.76) indigenous and 1/406 (0.25%) dog samples were seropositive, with 7/343 (2.04%) individuals the 1/144 (0.69%) dog from the Ocoy community, located in the city of São Miguel do Iguaçu, bordering Argentina at south, and far 10 km at west from Paraguay. All 84 healthcare professionals tested seronegative.

Biology of the non-parasitic phase of the cattle tick Rhipicephalus (Boophilus) microplus in an area of Amazon influence.

BACKGROUND: Rhipicephalus (Boophilus) microplus is the most important tick species affecting cattle in the world. Under field conditions, the non-parasitic phase of R. (B.) microplus is unknown in the Amazon biome, including Brazil. The present study aimed to evaluate the non-parasitic phase of R. (B.) microplus in field (grass plots) and laboratory conditions. METHODS: The study was conducted from September 2020 to April 2022 in an Amazonian region (Maranhão State, Brazil). We evaluated the biological parameters of R. (B.) microplus under laboratory and field conditions. Engorged females were exposed to experimental conditions every 14 days, totaling 20 months of study. The following biological parameters were observed: pre-oviposition period, egg mass incubation period, and maximum larval survival period. RESULTS: Abiotic data (e.g., temperature and humidity) varied little throughout the year. Precipitation was the factor that varied the most throughout the year (dry ~ 30 mm3 and rain 400 mm3), and the parameters of pre-oviposition and pre-hatching are longer during the rainy season. A possible negative effect of the dry season on the percentage of hatched eggs was observed. Larval longevity in the plots of both control and free females was short (mean ~ 50-60 days), below that recorded for larvae under controlled conditions (mean ~ 95 days). CONCLUSIONS: Rhipicephalus (Boophilus) microplus was able to complete its non-parasitic phase by producing host-seeking larvae in the pasture during all months of the study. The results indicate that R. (B.) microplus can complete up to six generations per year in biome Amazon. To our knowledge, this is the highest number of annual generations for R. (B.) microplus in Latin America.

New records of soft ticks (Acari: Argasidae) from caves in Brazil, with a morphological study of Ornithodoros fonsecai and an analysis of the taxonomic status of Antricola inexpectata.

In this study, we report soft ticks from bat-inhabiting caves in different areas of Brazil. From 2010 to 2019, we collected 807 tick specimens from nine caves located in four Brazilian states among two biomes. Ticks were morphologically identified as Antricola guglielmonei (282 specimens), Ornithodoros cavernicolous (260 specimens), and Ornithodoros fonsecai (265 specimens). Whereas A. guglielmonei was collected on bat guano in hot caves, O. cavernicolous and O. fonsecai were collected in cracks and crevices on the walls of cold caves, sometimes in the same chamber. Morphological identifications were corroborated by molecular and phylogenetic analyses inferred from tick mitochondrial 16S rRNA gene partial sequences. The sequences of A. guglielmonei, O. cavernicolous and O. fonsecai collected in this study clustered with conspecific GenBank sequences from different localities of Brazil. Remarkably, a clade containing 12 sequences of O. fonsecai was clearly bifurcated, denoting a degree of genetic divergence (up to 5 %) of specimens from Cerrado/Atlantic Forest biomes with the specimens from the Caatinga biome. To further evaluate this divergence, we performed morphometric analysis of the larval stage of different O. fonsencai populations by principal component analysis, which indicated that the larvae from Caatinga populations were generally smaller than the larvae from other biomes. Some of the present A. guglielmonei specimens were collected from the type locality of Antricola inexpectata. Comparisons of these specimens with the type specimens of A. inexpectata and A. guglielmonei indicated that they could not be separated by their external morphology. Hence, we are relegating A. inexpectata to a synonym of A. guglielmonei. This proposal is corroborated by our phylogenetic analysis.

Natural infection of free-ranging capybaras (Hydrochoerus hydrochaeris) with Anaplasmataecea and Rickettsiaceae bacteria in the Iberá wetlands ecoregion, Argentina.

The current work assessed the infection with Ehrlichia and Anaplasma species, and exposure to Rickettsia spp. in free-ranging capybaras in the Iberá wetlands ecoregion in Argentina. By indirect immunofluorescence assay, 37 out of 51 (73%) capybara sera were seropositive to Rickettsia spp., with 23.5% and 4% samples considered homologous to Rickettsia parkeri and Rickettsia bellii, respectively (or very closely related serotypes). Anaplasmataceae DNA was found to be highly prevalent in capybaras, with 33 out of 62 samples positive for Anaplasma sp. with Ct values of 28.64 ± 0.35 (average ± standard error), and 12 samples positive for Ehrlichia sp. with Ct values of 31.74 ± 0.87. Anaplasma sp. from capybaras was closely related to Anaplasma sp. reported to infect Amblyomma dubitatum in Iberá wetlands and to Anaplasma odocoilei, while the detected Ehrlichia sp. was closely related to "Candidatus Ehrlichia hydrochoerus" previously reported to infect capybaras in Brazil and A. dubitatum in Iberá wetlands. Structures compatible with Anaplasma morulae were observed in the cytoplasm of platelets from Anaplasma-positive capybaras. Our findings show that capybaras from the Iberá wetlands were exposed to Rickettsia species related to R. bellii and to the pathogen R. parkeri, and were infected with "Ca. Ehrlichia hydrochoerus" and a novel Anaplasma species, herein named "Candidatus Anaplasma capybara".

New tick records with notes on rickettsial infection from the wildlife of the state of Espírito Santo, southeastern Brazil.

This work aimed to report ticks infesting the wildlife among 15 municipalities of the state of Espírito Santo between 2016 and 2021, within the Atlantic Forest biome of southeastern Brazil. A total of 576 tick specimens (187 males, 56 females, 149 nymphs, and 184 larvae) was collected from 41 species of wild vertebrates (two reptiles, nine mammals, and 30 birds). Ticks were identified by morphological or molecular methods into 18 species, being 12, four, one and one of the genera Amblyomma, Ixodes, Rhipicephalus and Ornithodoros, respectively. Amblyomma rotundatum was the only species collected from reptiles. Ticks collected from mammals were identified as Amblyomma brasiliense, Amblyomma calcaratum, Amblyomma dubitatum, Amblyomma longirostre, Amblyomma nodosum, Amblyomma pacae, Amblyomma sculptum, Amblyomma varium and Rhipicephalus microplus. Amblyomma sculptum was the species found on the widest variety of hosts, collected from four mammal orders and five bird orders. Passeriformes birds were infested by Amblyomma fuscum, A. longirostre (also found on non- passerine birds), A. nodosum, Amblyomma parkeri, Amblyomma romarioi, A. varium and Ixodes loricatus. An adult female of Ixodes rio was collected from a Piciformes bird. Seabirds of the order Procellariiformes were infested by Ixodes percavatus sensu lato and Ixodes uriae. The argasid Ornithodoros capensis was collected from an offshore metallic platform that was used by Suliformes seabirds. Rickettsial agents of the spotted fever group, Rickettsia amblyommatis and Rickettsia sp. strain Pampulha, were detected in the ticks A. longirostre [from the Paraguayan hairy dwarf porcupine (Coendou spinosus)] and A. dubitatum [from the capybara (Hydrochoerus hydrochaeris)], respectively. The following nine tick species are reported for the first time in Espírito Santo state: A. calcaratum, A. fuscum, A. pacae, A. parkeri, A. romarioi, I. loricatus, I. rio, I. uriae, and O. capensis. Although it is also the first report of I. uriae in Brazil, we do not consider it established in the country. Multiple new tick-host associations are reported in the present study.

The anal pore route is efficient to infect Amblyomma spp. ticks with Rickettsia rickettsii and allows the assessment of the role played by infection control targets.

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.

Rickettsia communities and their relationship with tick species within and around the national park of Iguaçu, Brazil.

We report Rickettsia species from 2,334 ticks collected from environment (1,939 ticks) and animals (395 ticks) in the largest inland fragment of the Atlantic rainforest of southern Brazil and its fragments. Additionally, the DNA infection rates of Amblyomma ovale tick populations in the Neotropics with Rickettsia bellii and Rickettsia parkeri strain Atlantic rainforest were calculated using data from scientific publications, and their correlation was evaluated. From 11 tick species Rickettsia DNA was detected in seven (Amblyomma brasiliense, Amblyomma coelebs, Amblyomma incisum, Amblyomma longirostre, A. ovale, Haemaphysalis juxtakochi, Ixodes fuscipes) and was not detected in four species (Amblyomma dubitatum, Ixodes loricatus, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato). DNA of five Rickettsia species was detected (R. bellii, Rickettsia amblyommatis, Rickettsia rhipicephali, Rickettsia felis and Rickettsia sp. Aragaoi). To determine the prevalence of Rickettsia DNA positivity according to vector species, ticks were processed individually or in pools of 2-10 individuals (samples). The most prevalent Rickettsia species was R. bellii, found in 112 samples, followed by R. amblyommatis, R. rhipicephali, R. felis and Rickettsia sp. Aragaoi, found in 16, five, two and one sample, respectively. Rickettsia bellii DNA was found in five tick species with the highest infection rate in A. ovale and A. brasiliense. Absence of R. parkeri strain Atlantic rainforest in A. ovale ticks was an unexpected result. Furthermore, a negative correlation was identified between the infection rates (DNA) of R. bellii and/or R. parkeri strain Atlantic rainforest within A. ovale tick populations in the Neotropics. Putting together current knowledge, it can be proposed that, within natural settings, the diversity of rickettsiae and ticks creates a buffering effect on the overgrowth of rickettsiae and episodes of bacteremia in the hosts.

Molecular Detection of Tick-Borne Pathogens in Dogs from Indigenous Communities, Amazon, Brazil.

Background: There are few reports of tick-borne pathogens infecting dogs living in indigenous communities of Brazil. Herein, we aimed to molecularly detect vector-borne pathogens in dogs from two indigenous communities in the Brazilian Amazon. Materials and Methods: We surveyed 327 dogs raised in Amazon region at 2 distinct indigenous ethnicities for the molecular detection of tick-borne pathogens (114 from Tapirapé and 213 from Karajá indigenous ethnicity). Whole blood samples were subjected to PCR and sequencing for Ehrlichia, Babesia, and Hepatozoon. Univariate and multivariate analyses were used to investigate the factors affecting the pathogen infection patterns in dogs. Results: Among the 327 blood samples, 40 were positive for Ehrlichia canis (12.2%), 2 for Anaplasma platys (0.61%), and 204 were positive for Hepatozoon canis (66.5%). Binary Logistic Regression showed association between E. canis infection and ethnicity (p = 0.010) and tick attachment (p = 0.041). Karajá dogs were 3.4 times (95% CI 1.3-8.5) more likely to be positive for E. canis than Tapirapé dogs. Dogs with ticks were 2.5 times more likely (95% CI 1.0-7.6) to be positive for E. canis than dogs without ticks. Conclusions: Our survey expands the knowledge regarding the presence of vector-borne pathogens in dogs from indigenous communities in the Amazon region.

Description of Amblyomma monteiroae n. sp. (Acari: Ixodidae), a parasite of the great horned owl (Strigiformes: Strigidae) in southern Brazil.

In 2020, adult hard ticks (males and females) were collected from great horned owls [Bubo virginianus (Gmelin, 1788)] in the coastal region in southern Brazil. The engorged females were allowed to oviposit in the laboratory and hatched larvae could be obtained. Analyses of the external morphology of the adult ticks revealed that they represent a new species, which was named Amblyomma monteiroae n. sp. Partial sequences of the mitochondrial 16S rRNA gene and the nuclear second internal transcribed spacer (ITS2) were generated from a male and a female. Their 16S rRNA haplotypes were identical to each other and closest (96% identity) to corresponding sequences of Amblyomma parvitarsum Neumann, 1901, and 90% identical to Amblyomma neumanni Ribaga, 1902. Their ITS2 haplotypes were 95.8 to 96.0 identical to the single ITS-2 partial sequence of A. parvitarsum available in GenBank. In the phylogenetic trees inferred by both 16S rRNA and ITS2 partial sequences, A. monteiroae n. sp. formed a clade with A. parvitarsum, with A. neumanni branching sister to this clade. Amblyomma monteiroae n. sp. is genetically and morphologically related to A. parvitarsum. Both tick species are unique in combining the following morphological characters: scutum extensively ornate; eyes rounded and bulging; coxa I with two moderate pointed spurs, the external longer than the internal; a single triangular short spur on coxae II-III; presence of two spines on the tibia of legs II-IV; hypostomal dentition 3/3, trochanters without spurs. However, the males of the two species can be separated by specific features in palps and festoons, whereas the females differ in specific features of the coxal spurs. The larva of A. monteiroae n. sp. can be morphologically distinguished from A. parvitarsum only by morphometry, with the former species being slightly smaller. Currently, A. monteiroae n. sp. is restricted to southern Brazil, and the only known host is B. virginianus (Strigiformes: Strigidae). The present study increases the Amblyomma Brazilian fauna to 34 species.

Sharing of living area and exposure of domestic and wild canids to ticks, Rickettsia parkeri and Rangelia vitalii in the Atlantic Forest of southern Brazil.

Spotted fever caused by the bacterium Rickettsia parkeri, and canine rangeliosis caused by the protozoan Rangelia vitalii, are emerging or re-emerging tick-borne diseases in Brazil, where the main tick vectors are Amblyomma ovale and Amblyomma aureolatum, respectively. In the present study, we evaluated exposure to R. parkeri and R. vitalii infection among domestic (Canis lupus familiaris) and wild (Cerdocyon thous) canids sharing living areas within an Atlantic Forest area of southern Brazil. Four different domestic dogs and three different wild canids were captured. Most of the ticks collected from either domestic or wild canids were identified as A. ovale and A. aureolatum. DNA of R. parkeri was amplified from 14% of the A. ovale specimens. Two domestic dogs and two wild canids were seroreactive to R. parkeri antigens with high endpoint titers (>1024). Rangelia vitalii DNA was detected in two wild canids; one of them was resampled 93 days later, again infected by R. vitalii. We report exposure/infection of domestic dogs and wild canids to R. vitalii and/or R. parkeri-infected ticks in an Atlantic Forest area shared by both canid species, indicating that they also shared the same populations of the tick vectors, A. aureolatum and A. ovale. While A. ovale, A. aureolatum, R. parkeri, R. vitalii and C. thous, are all native to the Atlantic Forest, further studies are needed to evaluate the impact of an exotic canine species - C. lupus familiaris - on the enzootic cycles of R. vitalii and R. parkeri, as well as the possible role of domestic dogs in emergence and re-emergence of R. parkeri-spotted fever in humans and canine rangeliosis, respectively.

Description of a new Pavlovskyella species (Acari: Argasidae) from Chile.

Soft ticks (Argasidae) of the Pavlovskyella Pospelova-Shtrom subgenus are important vectors of relapsing fever spirochetes, which are agents of disease globally. South American representatives of the Pavlovskyella subgenus include 3 species: Ornithodoros (Pavlovskyella) brasiliensis Aragão, Ornithodoros (Pavlovskyella) furcosus Neumann, and Ornithodoros (Pavlovskyella) rostratus Aragão. Here, we describe a fourth species based on morphological and mitogenomic evidence of ticks collected in burrows of unknown hosts in central Chile. The larva of the new species separates from other South American soft ticks by the following combination of characters: 13 pairs of dorsolateral setae, dorsal plate hexagonal, hypostome blunt with denticles from apex almost to the base. Adults of this new species lack cheeks, possess a dorsoventral groove, and have humps, similar to O. (P.) brasiliensis; however, they lack bulging structures on the flanks of idiosoma. Moreover, females and males differ from O. (P.) rostratus by having 3 humps instead of spurs in tarsi I and from O. (P.) furcosus because of their smaller size and thinner anterior lip of the genital aperture in females. The phylogenetic analysis performed with mitogenomes of the Argasidae family depicts the new Pavlovskyella species from Chile in a monophyletic clade with other South American species in the subgenus, confirming a regional group.

Morphological anomalies in hard ticks (Acari: Ixodidae) from Brazil.

Tick abnormalities have been unusual in nature, and they can be divided into local and general. In the present study, external morphological anomalies were described in 31 individual adult ticks of 15 different species of Ixodidae, which were collected on wild hosts (20 ticks), domestic hosts (7 ticks), and in the environment (4 ticks) in 11 states of Brazil from 1998 to 2022. Among the 31 tick specimens, 14 (45%) were categorized as local anomalies, and 17 (55%) as general anomalies. The ticks were taxonomically identified into 14 species of Amblyomma, and one species of Rhipicephalus. Local anomalies included malformations of scutum/alloscutum, ectromely, leg atrophy, and a third ectopic spiracular plate. General anomalies included opisthosoma duplication, no expansion of dorsal alloscutum in engorged females, and gynandromorphism; the latter is described for 13 tick specimens. Morphological anomalies in Amblyomma aureolatum, Amblyomma brasiliense, Amblyomma humerale and Amblyomma longirostre are reported for the first time. Although the results herein expand the list of anomalous tick species in the Neotropics, future studies should be conducted to clarify the origin of these anomalies.

Ticks and tick-borne rickettsiae from dogs in El Salvador, with report of the human pathogen Rickettsia parkeri.

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.

Detection of Rickettsia spp. in Animals and Ticks in Midwestern Brazil, Where Human Cases of Rickettsiosis Were Reported.

Brazilian spotted fever (BSF) is the most important tick-borne diseases affecting humans in Brazil. Cases of BSF have recently been reported in the Goiás state, midwestern Brazil. All cases have been confirmed by reference laboratories by seroconversion to Rickettsia rickettsii antigens. Because serological cross-reactions among different rickettsial species that belong to the spotted fever group (SFG) are common, the agent responsible for BSF cases in Goiás remains unknown. From March 2020 to April 2022, ticks and plasma were collected from dogs, horses and capybaras (Hydrochoerus hydrochaeris), and from the vegetation in an area where BSF cases have been reported and two areas under epidemiological surveillance in Goiás. Horses were infested by Amblyomma sculptum, Dermacentor nitens and Rhipicephalus microplus; dogs by Rhipicephalus sanguineus sensu lato (s.l.), Amblyomma ovale and A. sculptum, and capybaras by A. sculptum and Amblyomma dubitatum. Adults of A. sculptum, A. dubitatum, Amblyomma rotundatum and immature stages of A. sculptum and A. dubitatum, and Amblyomma spp. were collected from the vegetation. DNA of Rickettsia that did not belong to the SFG was detected in A. dubitatum, which was identified by DNA sequencing as Rickettsia bellii. Seroreactivity to SFG and Rickettsia bellii antigens was detected in 25.4% (42/165) of dogs, 22.7% (10/44) of horses and 41.2% (7/17) of capybaras, with higher titers for R. bellii in dogs and capybaras. The seropositivity of animals to SFG Rickettsia spp. antigens demonstrates the circulation of SFG rickettsiae in the region. Further research is needed to fully determine the agent responsible for rickettsiosis cases in this area.

'Candidatus Rickettsia andeanae' and Probable Exclusion of Rickettsia parkeri in Ticks from Dogs in a Natural Area of the Pampa Biome in Brazil.

Spotted fever illness caused by the tick-borne pathogen Rickettsia parkeri has emerged in the Pampa biome in southern Brazil, where the tick Amblyomma tigrinum is implicated as the main vector. Because domestic dogs are commonly parasitized by A. tigrinum, this canid is also a suitable sentinel for R. parkeri-associated spotted fever. Herein, we investigate rickettsial infection in ticks, domestic dogs and small mammals in a natural reserve of the Pampa biome in southern Brazil. The ticks A. tigrinum, Amblyomma aureolatum and Rhipicephalus sanguineus were collected from dogs. Molecular analyses of ticks did not detect R. parkeri; however, at least 34% (21/61) of the A. tigrinum ticks were infected by the non-pathogenic agent 'Candidatus Rickettsia andeanae'. Serological analyses revealed that only 14% and 3% of 36 dogs and 34 small mammals, respectively, were exposed to rickettsial antigens. These results indicate that the study area is not endemic for R. parkeri rickettsiosis. We tabulated 10 studies that reported rickettsial infection in A. tigrinum populations from South America. There was a strong negative correlation between the infection rates by R. parkeri and 'Candidatus R. andeanae' in A. tigrinum populations. We propose that high infection rates by 'Candidatus R. andeanae' might promote the exclusion of R. parkeri from A. tigrinum populations. The mechanisms for such exclusion are yet to be elucidated.

The survival of Amblyomma sculptum ticks upon blood-feeding depends on the expression of an inhibitor of apoptosis protein.

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.

A Novel Relapsing Fever Group Borrelia Isolated from Ornithodoros Ticks of the Brazilian Caatinga.

Tick-borne relapsing fever group (RFG) borreliosis remains neglected as a human disease and little is known on its maintenance in ticks and vertebrates, especially in South America. Therefore, this study investigated borrelial infection in Ornithodoros ticks collected in rodent-inhabited rock formations in the Brazilian semiarid region, within the Caatinga biome. Collected ticks (Ornithodoros rietcorreai and Ornithodoros cf. tabajara) were allowed to feed under laboratory conditions on guinea pigs, which had blood samples examined daily by dark-field microscopy. No spirochetes were visualized in the blood of any of four O. rietcorreai-infested guinea pigs. Contrastingly, spirochetes were visualized between 9 and 39 days after tick feeding in the blood of three guinea pigs, each infested with O. cf. tabajara ticks from a different locality. Guinea pig infection was confirmed by passages into experimental animals and by generating DNA sequences of Borrelia spp. from the blood of spirochetemic guinea pigs. Three O. cf. tabajara populations were infected by the same borrelial organism, which was characterized as a novel RFG agent (named as 'Candidatus Borrelia caatinga') based on 10 Borrelia loci (rrs, flaB, glpQ, gyrB, clpX, pepX, pyrG, recG, rplB and uvrA). We demonstrated that O. cf. tabajara is a competent vector of the novel Borrelia sp. isolates, although none of the infected rodents developed clinical illness.

Morphological and molecular confirmation of Ornithodoros hasei (Schulze, 1935) (Acari: Argasidae) in Colombia.

A large number of tick species are proven vectors for the transmission of bacteria, protozoa, and viruses. Soft ticks (Acari: Argasidae) in South America have been found to be the most frequent carriers of borreliae of the relapsing fever group (RFG); however, there are several information gaps specially on the taxonomy and distribution of some tick species. Here, we used light microscopy, scanning electron microscopy, and PCR amplification of a fragment of the mitochondrial 16S rRNA gene to evaluate 174 larvae of Ornithodoros (Argasidae) collected from three bat species (Eptesicus orinocensis, Molossus rufus and Noctilio albiventris) in the Orinoquia Region of Colombia. The morphological and molecular results confirmed that all the analyzed larvae corresponded to Ornithodoros hasei. Comparisons of mitochondrial 16S rDNA sequences showed low genetic divergence (0% - 0.3%) between larvae of the Department of Arauca in the Orinoquia Region and higher genetic divergence (3.4 - 4.7%) in sequences from other American countries. Our work represents the most recent collection of this species in Colombia and provides a molecular evaluation for the first time. Moreover, a new association of O. hasei with bats such as E. orinocensis is documented. Considering the wide distribution of O. hasei in the American Continent, and its putative role as vector for Borrelia, integrative studies that involve morphological, morphometric, molecular data and experimental crosses are needed to determine if the higher genetic distances are associated with cryptic speciation, as detected in other tick complexes, or represent genetic divergences among geographically different populations of O. hasei.