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This study was carried out from February 2020 to September 2021 in Parque Nacional das Emas (PNE), a national park located in the Cerrado biome, midwestern Brazil, as well as in surrounding rural properties. Serum and tick samples were collected from dogs, terrestrial small mammals, and humans. Ticks were also collected from the environment. Dogs were infested with Rhipicephalus linnaei adults, whereas small mammals were infested by immature stages of Amblyomma spp., Amblyomma triste, Amblyomma dubitatum, and Amblyomma coelebs. Ticks collected from vegetation belonged to several species of the genus Amblyomma, including A. coelebs, A. dubitatum, Amblyomma naponense, Amblyomma sculptum, and A. triste. Two Rickettsia species were molecularly detected in ticks: Rickettsia parkeri in A. triste from the vegetation and a Rickettsia sp. (designated Rickettsia sp. strain PNE) in A. sculptum and A. triste collected from lowland tapirs (Tapirus terrestris). Based on short gltA gene fragments, this rickettsial organism showed 99.7–100% to Rickettsia tillamookensis. Seroreactivity to Rickettsia antigens was detected in 21.9% of dogs, 15.4% of small mammals, and 23.5% of humans. The present study reveals the richness of ticks and demonstrates the circulation of rickettsial agents in one of the largest conservation units in the Cerrado biome in Brazil. To our knowledge, this is the first report of a rickettsial phylogenetically related to R. tillamookensis in Brazil. [ABSTRACT FROM AUTHOR]

Amblyomma sculptum is a species of public health interest because it is associated with the transmission of the bacteria that causes Brazilian Spotted Fever (BSF). The use of repellents on humans is a prophylactic measure widely used to provide protection against a series of arthropod vectors, including mosquitoes and ticks. However, in Brazil, the effectiveness of commercial repellents against A. sculptum is little known. Therefore, it is necessary to carry out specific studies to evaluate the repellency of these commercial products, registered for use against mosquitoes, against the star tick. The main goal of the present work was to evaluate the effectiveness of six commercial repellents against A. sculptum. Unfed nymphs, aged between two and eight weeks, were tested against products based on DEET (OFF!® and Repelex®), Icaridin (Exposis® and SBP®), and IR3535 (Johnsons and Henlau). Three bioassays were conducted to evaluate tick behavior: (i) filter paper, (ii) fingertip, and (iii) field. All bases tested showed high repellent activity, differing significantly (p < 0.05) from the control. It was observed the formulation with DEET resulted in the best results in the filter paper bioassay. In the fingertip bioassay, the DEET base repelled a greater number of ticks compared to Icaridin. In the field bioassay, there was no significant difference between the Icaridin base and DEET, and both formulations differed from the control (p < 0.05). The six formulations tested showed significant percentages of repellency against ticks; however, for the fingertip and field bioassays, the products OFF!®, Repelex®, and Exposis® were tested as they showed better performance in the filter paper test. OFF!® showed the best percentage of repellency (100%), followed by Repelex® (96.8%), and Exposis® (93.1%), considering the two-hour period of the bioassay-field-test. Proving the effectiveness of repellents on the market against A. sculptum presented in this study is crucial, since this is the main ectoparasite of humans that can transmit Rickettsia rickettsii when infected. The effectiveness of commercial insect repellents against other tick species that parasitize humans can also be explored. [ABSTRACT FROM AUTHOR]

This work aimed to determine if the tick species, Amblyomma sculptum and Amblyomma tonelliae, hybridize along their contact zones in Argentina. Free‐living adults and nymphs of A. sculptum and A. tonelliae were collected in seven sampling locations of northern Argentina. In four of them, the two species occur in parapatry (possible hybrid zone) whereas in the other three sites, only one species is known to occur. A total of 65 A. sculptum and 65 A. tonelliae from both, allopatric and parapatric populations, were analysed. The nuclear (ITS2) and mitochondrial (COI and 12SrDNA) gene sequences of each tick were amplified and analysed to verify whether or not they could reveal the presence of hybrids among the parapatric samples. No morphological and molecular evidence was found to support the hypothesis of ongoing natural hybridization. Intrinsic postzygotic barriers may be the cause of lack of gene flow between the two species in areas of co‐ocurrence. The results can be explained by the length of time the two lineages spent in allopatry since the middle of the Miocene and before their respective distribution range expanded again reaching a narrow secondary contact zone. [ABSTRACT FROM AUTHOR]

Ticks have saliva rich in immunoregulatory molecules that interfere with the host's physiology in order to feed. This study aimed to evaluate the concentration of acute phase proteins and circulating oxidative stress in response to infestation by Amblyomma sculptum and Dermacentor nitens in two breed horses, Mangalarga Marchador and Breton Postier, to define resistance or susceptibility to ticks. Among the oxidative stress markers, we observed lower malondialdehyde and nitric oxide in horses with tick infestation, consequently not altering the antioxidant enzymes. Breton Postier with tick infestation showed a reduction in the ferric reducing ability of plasma (FRAP), which may be due to lower feeding of the host due to the stress caused by the infestation or even to sequestration of components induced by the tick during blood feeding. The alpha-1-antitrypsin, an acute phase protein, showed an increase in Mangalarga Marchador with tick infestation; curiously it is related to a protective action against tissue damage, pathogens and parasites. We could assume that Mangalarga Marchador showed a better response to ticks when compared to Breton Postier. However, it is still early to define the resistance or susceptibility to ticks, as we did not observe significant changes in most of the analyzed variables. Further studies are needed to understand the compounds and mechanisms of action of the tick saliva in the acute phase proteins and the possible relationships of oxidative stress in the host and the tick during blood feeding. [ABSTRACT FROM AUTHOR]

Procyonids are reservoirs of many zoonotic infectious diseases, including tick-borne pathogens. The role of coatis (Nasua nasua) in the epidemiology of piroplasmids and Rickettsia has not been fully addressed in Brazil. To molecularly study these agents in coatis and associated ticks, animals were sampled in two urban areas in Midwestern Brazil. Blood (n = 163) and tick (n = 248) DNA samples were screened by PCR assays targeting the 18S rRNA and gltA genes of piroplasmids and Rickettsia spp., respectively. Positive samples were further molecularly tested targeting cox-1, cox-3, β-tubulin, cytB, and hsp70 (piroplasmid) and ompA, ompB, and htrA 17-kDa (Rickettsia spp.) genes, sequenced and phylogenetically analyzed. All coatis' blood samples were negative for piroplasmids, whereas five pools of ticks (2%) were positive for two different sequences of Babesia spp.. The first from Amblyomma sculptum nymphs was close (i.e., ≥ 99% nucleotide identity) to a Babesia sp. previously found in capybaras (Hydrochoerus hydrochaeris); the second from Amblyomma dubitatum nymphs and Amblyomma spp. larvae was identical (100% nucleotide identity) to a Babesia sp. detected in opossums (Didelphis albiventris) and associated ticks. Four samples (0.8%) were positive by PCR to two different Rickettsia spp. sequences, being the first from Amblyomma sp. larva identical to Rickettsia belli and the second from A. dubitatum nymph identical to Rickettsia species from Spotted Fever Group (SFG). The detection of piroplasmids and SFG Rickettsia sp. highlights the importance of Amblyomma spp. in the maintenance of tick-borne agents in urban parks where humans and wild and domestic animals are living in sympatry. [ABSTRACT FROM AUTHOR]

Simple Summary: Capybaras are known hosts for various tick species, but there are limited data regarding the tick-borne pathogens they can carry. We assessed the presence of piroplasmids and Ehrlichia spp. in capybaras and their associated ticks in Goiás state, central-western Brazil. Neither capybaras nor ticks were positive for Ehrlichia spp. However, we detected an undescribed species of protozoan in both the capybaras and ticks. Further research is required for a formal delineation of this protozoan species, as well as to investigate the role of these ticks as vectors and the possible pathogenicity of this parasite to other animals, including horses. Capybaras (Hydrochoerus hydrochaeris) are the largest rodents on Earth. While capybaras are hosts for various tick species, there is limited information regarding the tick-borne pathogens they can carry. We investigated the presence of piroplasmids and Ehrlichia spp. in capybaras and their associated ticks in two peri-urban areas in Goiás state, central-western Brazil. Blood samples collected from 23 capybaras were used to investigate the presence of piroplasmids and Ehrlichia spp. in stained-blood smears and by PCR. Ticks collected from the capybaras were identified morphologically and also tested using PCR for the same pathogens. A total of 955 ticks were collected, including 822 (86.1%) Amblyomma sculptum, 132 (13.8%) Amblyomma dubitatum, and one (0.1%) unidentified larva of Amblyomma sp. Neither the capybaras nor ticks were positive for Ehrlichia spp. However, a stained-blood smear examination revealed the presence of ring-stage and pyriform-shaped merozoites in the erythrocytes of one (4.4%) capybara. In the same way, 47.8% (11/23) and 19.9% (36/181) of blood samples and ticks, respectively, were positive for piroplasmids in the PCR. We successfully sequenced a partial 18S rRNA gene fragment of four samples (two capybaras, one A. sculptum, and one A. dubitatum), and the phylogenetic reconstruction disclosed that the organism reported in the present study clusters within the genus Babesia. Further research is required for a formal delineation of this species (designated as Babesia sp. strain Capybara) and to investigate the hypothesis of A. dubitatum and A. sculptum ticks being vectors. [ABSTRACT FROM AUTHOR]

Rickettsia rickettsii infection causes the highest human fatality rate among all Rickettsia species of the world and is endemic in Southeast Brazil. In this part of the country most human spotted fever cases are related to unnaturally high populations of ticks, usually a mix of two species, Amblyomma dubitatum Neumann and Amblyomma sculptum Berlese and their local host, capybara (Hydrochoerus hydrochaeris). At the same time, an increase in the number of SF notifications as well as its urbanization was observed, and a better characterization of disease epidemiology is mandatory for control measures and to halt its expansion. It was recently noticed in southeast Brazil that SF endemic areas were characterized by overgrowth populations of A. sculptum and decreased populations of A. dubitatum. Hence, characterization of areas with potential to endemicity, eco-epidemiological studies and control measures may rely on the evaluation of A. sculptum/A. dubitatum rate. However, in potentially endemic scenarios, discriminating the two tick species, particularly the nymphs considered the main vector stage, is a challenge in face of several hundreds to thousands of ticks that should be examined for quantitative studies. We herein present additional morphological features to an existing identification key for Amblyomma nymphs that considerably diminishes the labor to distinguish nymphs of these two tick species. Specifically, the oval-shaped scutum of A. dubitatum and a conspicuous scapula of A. sculptum are prominent discriminating features. [ABSTRACT FROM AUTHOR]

Amblyomma sculptum is the main tick associated with human bites in Brazil and the main vector of Rickettsia rickettsii , the causative agent of the most severe form of Brazilian spotted fever. Molecules produced in the salivary glands are directly related to feeding success and vector competence. In the present study, we identified sequences of A. sculptum salivary proteins that may be involved in hematophagy and selected three proteins that underwent functional characterization and evaluation as vaccine antigens. Among the three proteins selected, one contained a Kunitz_bovine pancreatic trypsin inhibitor domain (named AsKunitz) and the other two belonged to the 8.9 kDa and basic tail families of tick salivary proteins (named As8.9kDa and AsBasicTail). Expression of the messenger RNA (mRNA) encoding all three proteins was detected in the larvae, nymphs, and females at basal levels in unfed ticks and the expression levels increased after the start of feeding. Recombinant proteins rAs8.9kDa and rAsBasicTail inhibited the enzymatic activity of factor Xa, thrombin, and trypsin, whereas rAsKunitz inhibited only thrombin activity. All three recombinant proteins inhibited the hemolysis of both the classical and alternative pathways; this is the first description of tick members of the Kunitz and 8.9kDa families being inhibitors of the classical complement pathway. Mice immunization with recombinant proteins caused efficacies against A. sculptum females from 59.4% with rAsBasicTail immunization to more than 85% by immunization with rAsKunitz and rAs8.9kDa. The mortality of nymphs fed on immunized mice reached 70–100%. Therefore, all three proteins are potential antigens with the possibility of becoming a new tool in the control of A. sculptum. [ABSTRACT FROM AUTHOR]

The bacterium Rickettsia rickettsii causes Brazilian spotted fever (BSF), a highly lethal disease that is transmitted by Amblyomma sculptum ticks in areas where capybaras (Hydrochoerus hydrochaeris) are the tick's major hosts. In this study, we evaluated the expansion of a capybara population in a residential park in São Paulo state, and the implications of such expansion to the occurrence of ticks and BSF. The capybara population was quantified during 2004–2013. In 2012, there was a BSF human case in the area, culminating in the complete fencing of the residential park and the official culling of all capybaras. Quantification of ticks in the environment was performed by dry ice traps from 2005 to 2018. Domestic dogs in 2006–2011 and capybaras in 2012 were serologically tested for the presence of anti-R. rickettsii antibodies. Our results show that capybara numbers increased ≈5 times from 2004 (41 capybaras) to 2012 (230 capybaras). Dry ice traps collected A. sculptum and Amblyomma dubitatum. The number of A. dubitatum adult ticks was generally higher than A. sculptum adults during 2005–2006; however, during 2012–2013, A. sculptum outnumbered A. dubitatum by a large difference. During 2016–2018 (after capybara culling), the number of both species fell close to zero. The low numbers of A. sculptum adult ticks during 2005–2006 coincided with relatively low capybara numbers (<80). Thereafter, in 2012, we counted the highest numbers of both A. sculptum ticks and capybaras (230 animals). All 40 canine blood samples were seronegative to R. rickettsii, in contrast to the 48.3% seropositivity (83/172) among capybaras. Our results support that the emergence of BSF in the residential park was a consequence of the increase of the local capybara population, which in turn, provided the increment of the A. sculptum population. Culling the entire capybara population eliminated the risks of new BSF cases. [ABSTRACT FROM AUTHOR]

Amblyomma sculptum is an important vector of Rickettsia rickettsii , causative agent of Rocky Mountain spotted fever and the most lethal tick-borne pathogen affecting humans. To feed on the vertebrate host's blood, A. sculptum secretes a salivary mixture, which may interact with skin resident dendritic cells (DCs) and modulate their function. The present work was aimed at depicting the A. sculptum saliva-host DC network and the biochemical nature of the immunomodulatory component(s) involved in this interface. A. sculptum saliva inhibits the production of inflammatory cytokines by murine DCs stimulated with LPS. The fractionation of the low molecular weight salivary content by reversed-phase chromatography revealed active fractions eluting from 49 to 55% of the acetonitrile gradient. Previous studies suggested that this pattern of elution matches with that observed for prostaglandin E2 (PGE2) and the molecular identity of this lipid mediator was unambiguously confirmed by a new high-resolution mass spectrometry methodology. A productive infection of murine DCs by R. rickettsii was demonstrated for the first time leading to proinflammatory cytokine production that was inhibited by both A. sculptum saliva and PGE2, a result also achieved with human DCs. The adoptive transfer of murine DCs incubated with R. rickettsii followed by treatment with A. sculptum saliva or PGE2 did not change the cytokine profile associated to cellular recall responses while IgG2a-specific antibodies were decreased in the serum of these mice. Together, these findings emphasize the role of PGE2 as a universal immunomodulator of tick saliva. In addition, it contributes to new approaches to explore R. rickettsii -DC interactions both in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Ticks endure stressful off-host periods and perform as vectors of a diversity of infectious agents, thus engaging pathways that expectedly demand for autophagy. Little is known of ticks' autophagy, a conserved eukaryotic machinery assisting in homeostasis processes that also participates in tissue-dependent metabolic functions. Here, the autophagy-related ATG4 (autophagin-1), ATG6 (beclin-1) and ATG8 (LC3) mRNAs from the human diseases vector Amblyomma sculptum and the cattle-tick Rhipicephalus microplus were identified. Comparative qPCR quantifications evidenced different transcriptional status for the ATG genes in the salivary glands (SG), ovaries and intestines of actively feeding ticks. These ATGs had increased relative transcription under nutrient-deprivation, as determined by validation tests with R. microplus embryo-derivative cells BME26 and A. sculptum SG explants incubations in HBSS. Starvation lead to 4-31.8× and ~ 60-500× increments on the ATGs mRNA loads in BME26 and A. sculptum SG explants, respectively. PI3K inhibitor 3MA treatment also affected ATGs expression in BME26. Some ATGs were more transcribed in the SGs than in the ovaries of cattle-ticks. Amblyomma sculptum/ R. microplus interspecific comparisons showed that ATG4 and ATG6 were 0.18× less expressed in A. sculptum SGs, but ~ 10-100× more expressed in their ovaries when compared to R. microplus organs. ATG4 and ATG8a transcript loads were ~ 120× and ~ 40× higher, respectively, in A. sculptum intestines when compared to cattle-ticks of similar weight category. ATGs expression in A. sculptum intestines increased with tick weight, indicating Atgs contribution to intracellular blood digestion. Possible roles of the autophagy machinery and their organ-specific expression profile on vector biology are discussed. [ABSTRACT FROM AUTHOR]

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Background: Until recently, Amblyomma cajennense (Fabricius, 1787) was considered to represent a single tick species in the New World. Recent studies have split this taxon into six species. While the A. cajennense species complex or A. cajennense (sensu lato) (s.l.) is currently represented by two species in Brazil, A. cajennense (sensu stricto) (s.s.) and Amblyomma sculptum Berlese, 1888, their geographical distribution is poorly known. Methods: The distribution of the A. cajennense (s.l.) in Brazil was determined by morphological examination of all lots of A. cajennense (s.l.) in two large tick collections of Brazil, and by collecting new material during three field expeditions in the possible transition areas between the distribution ranges of A. cajennense (s.s.) and A. sculptum. Phylogenetic analysis inferred from the ITS2 rRNA gene was used to validate morphological results. Morphological description of the nymphal stage of A. cajennense (s.s.) is provided based on laboratory-reared specimens. Results: From the tick collections, a total 12,512 adult ticks were examined and identified as 312 A. cajennense (s.s.), 6,252 A. sculptum and 5,948 A. cajennense (s.l.). A total of 1,746 ticks from 77 localities were collected during field expeditions, and were identified as 249 A. cajennense (s.s.), 443 A. sculptum, and 1,054 A. cajennense (s.l.) [these A. cajennense (s.l.) ticks were considered to be males of either A. cajennense (s.s.) or A. sculptum]. At least 23 localities contained the presence of both A. cajennense (s.s.) and A. sculptum in sympatry. DNA sequences of the ITS2 gene of 50 ticks from 30 localities confirmed the results of the morphological analyses. The nymph of A. cajennense (s.s.) is morphologically very similar to A. sculptum. Conclusion: Our results confirmed that A. cajennense (s.l.) is currently represented in Brazil by only two species, A. cajennense (s.s.) and A. sculptum. While these species have distinct distribution areas in the country, they are found in sympatry in some transition areas. The current distribution of A. cajennense (s.l.) has important implications to public health, since in Brazil A. sculptum is the most important vector of the bacterium Rickettsia rickettsii, the etiological agent of Brazilian spotted fever. [ABSTRACT FROM AUTHOR]

Pantanal is a huge floodplain mostly in Brazil, and its main economic activity is extensive cattle raising, in farms characterized by an extremely wildlife-rich environment. We herein describe tick infestations of cattle and of the natural environment in Pantanal of Nhecolândia in Brazil, at areas with and without cattle during both dry and wet seasons. Environmental sampling resulted in three tick species: Amblyomma sculptum (423 nymphs and 518 adults), Amblyomma parvum (7 nymphs and 129 adults), Amblyomma ovale (3 adults) as well as three clusters and two individuals of Amblyomma sp. larvae. A significantly higher number of adult A. sculptum ticks was found in areas with cattle in the wet season. From 106 examinations of bovines 1710 ticks from three species were collected: Rhipicephalus microplus (55.7 % of the total), A. sculptum (38 %) and A. parvum (4.1 %), as well as 32 Amblyomma sp. larvae. A significant similarity was found between Amblyomma tick fauna from environment and on cattle during both seasons. All A. sculptum females on bovines were flat whereas many of A. parvum females and A. sculptum nymphs were engorging. Although R. microplus was the most abundant tick species on cattle, overall highest tick prevalence on bovines in the dry season was of A. sculptum nymphs. Lack of R. microplus in environmental sampling, relationship between cattle and increase in adult A. sculptum numbers in the environment as well as suitability of bovine for the various tick species are discussed. [ABSTRACT FROM AUTHOR]

The Pantanal in Brazil is the largest floodplain of the world. This ecosystem, rich in wildlife, has a large feral pig population. Such a large host biomass must have a strong influence on the parasite fauna. In this work, we evaluated the role of feral pigs in the maintenance of Amblyomma sculptum (formerly Amblyomma cajennense), the most prevalent tick species in the Pantanal. Tick infestations were evaluated on 243 feral pigs and their environment. The suitability of domestic pigs, representing their feral relatives, to A. sculptum adults and nymphs was assessed experimentally. Tick infestation of feral pigs was strongly associated with that of the environment: 96 and 97 % of the ticks, respectively, were A. sculptum. The infestation prevalence on this host species was close to 90 % in the dry season and 100 % in the wet season and mean infestation intensity was above 30 ticks in both seasons. Suitability of pigs as hosts for A. sculptum was shown by the high proportion of nymphs and female ticks found engorging on captured feral pigs and adequate biological parameters displayed by ticks from experimental infestations of domestic pigs. Other tick species on feral pigs, albeit in much lower numbers, were Amblyomma parvum and Ornithodorus rostratus. Results show that feral pigs feed a high proportion of the A. sculptum adults and nymphs in their territories and should be a target for tick-borne diseases studies. This is particularly relevant to public health because all the main tick species found on feral pigs are aggressive to humans as well. [ABSTRACT FROM AUTHOR]

Simple Summary: The knowledge of the dynamics of ticks in wild animals is essential for surveillance of tick-borne diseases. Coatis (Nasua nasua) are mammals that easily adapt to anthropized areas, favoring close contact with domestic animals and humans, favoring the exchange of ticks and tick-borne agents. The present study aimed to investigate the tick diversity on coatis from forest urban areas of midwestern Brazil, as well as the dynamics of ticks during the seasons of the year and the correlation between tick species and gender and age of the sampled coatis. Three tick species were identified parasitizing coatis from forested urban fragments, namely A. dubitatum nymphs, A. sculptum adults and nymphs, and A. ovale adults. After analyzing the obtained results, it is likely that coatis from anthropized areas present tick species diversity lower than those from natural landscapes. The mean intensity and prevalence of Amblyomma larvae and nymphs is similar among males and females as well as in immature and mature animals, which might reflect the gregarious behavior of coatis, since adult males live together with females and offspring outside and inside the mating season, forming large groups of individuals. Understanding the diversity and ecology of ectoparasites in wild animals is essential for surveillance of vector-borne diseases. Coatis (Nasua nasua) easily adapt to anthropized areas, favoring close contact with domestic animals and humans, with the possibility of exchange of ectoparasites and pathogens. The present study aimed to identify the diversity of ticks parasitizing coatis from forest urban areas of midwestern Brazil, to evaluate the seasonal dynamics of ticks during the seasons of the year, and to assess the correlation between tick species and gender and age of the sampled coatis. For this purpose, 103 coatis were captured in two Conservation areas, both located in Campo Grande city, Mato Grosso do Sul state, Midwestern Brazil. The animals' entire body was inspected for the presence of ectoparasites, and ticks were removed for taxonomic identification. In total, 168 captures were performed in both areas during the observational study considering the first capture and recaptures. In total, 2242 ticks were collected: 838 Amblyomma larvae, 1241 A. sculptum nymphs, and 150 A. dubitatum nymphs. Thirteen adult ticks were identified as three males and five females of A. sculptum and two males and three females of A. ovale. While a quantity of Amblyomma larvae was observed in the first months of the year (January, April and May), Amblyomma nymphs showed a higher quantity during the months of July, August, October and November. No statistical difference was observed when comparing mean intensity and prevalence of Amblyomma larvae, nymphs of A. sculptum and A. dubitatum between the two sampled areas, males vs. females and immature vs. mature animals. In conclusion, three tick species were identified parasitizing coatis from forested urban fragments in midwestern Brazil, namely A. dubitatum nymphs, A. sculptum adults and nymphs, and A. ovale adults. Coatis from anthropized areas seem to present tick species diversity lower than those from natural areas. The lack of statistical difference regarding mean intensity and prevalence of Amblyomma larvae and nymphs between males vs. females and immature vs. mature animals might have reflected the gregarious behavior of coatis, since adult males live together with females and offspring outside and inside the mating season, forming large groups of individuals. [ABSTRACT FROM AUTHOR]

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