Distribution modeling of Amblyomma rotundatum and Amblyomma dissimile in Brazil: estimates of environmental suitability.

The number of reports of tick parasitism in amphibians and reptiles has increased over the past few years, including new host and location records for Amblyomma rotundatum and Amblyomma dissimile. However, knowledge of the geographical distribution remains incomplete, and in many regions of Brazil, the presence of these vectors has not been investigated. Several candidate models were built using a correlative maximum entropy approach, and best-fitting models were selected based on statistical significance, predictive ability, and complexity based on current climatic trends and future projected climate changes. Final models showed a good ability to discriminate A. rotundatum and A. dissimile current and future potential distributions. The entire country had higher predicted suitability for A. rotundatum while A. dissimile was mainly restricted to the Amazon and Pantanal biomes. A. rotundatum is a species with enormous potential for dissemination in the next decades, potentially through the legal and illegal transport of reptiles and amphibians. The proposed model is useful for targeting surveillance efforts increasing the efficiency and accuracy of future ecological research and tick management efforts.

"Candidatus Borrelia ibitipoquensis," a Borrelia valaisiana-Related Genospecies Characterized from Ixodes paranaensis in Brazil.

Borrelia burgdorferi sensu lato (Bbsl) spirochetes include the agents of Lyme borreliosis in temperate regions of the Northern Hemisphere, and merge their transmission cycles mainly with ticks of the Ixodes ricinus complex. Twenty genospecies compose Bbsl currently, and with the exception of Borrelia chilensis, and Borrelia garinii, all have been described only for North America, Europe, North Africa, and Asia. Here, we collected specimens of Ixodes paranaensis, a tick associated with swifts in a Brazilian natural park from the state of Minas Gerais, and performed a molecular characterization of 11 borrelial genes. Based on comparisons of inter and intraspecific genetic divergences, and Bayesian phylogenetic trees inferred for 16S rRNA, flaB, p66, and concatenated clpA, clpX, pepX, pyrG, recG, nifS rlpB, and uvrA genes, we demonstrate the occurrence of a new genospecies of Bbsl. "Candidatus Borrelia ibitipoquensis" Ip37 is closely related to Borrelia sp. Am501, and Borrelia valaisiana, a spirochete transmitted by ticks of the I. ricinus complex in Eurasia that uses birds as reservoirs. In a similar ecological scenario involving ticks and avian hosts, the migratory swift Streptoprocne biscutata is the sole-documented bird associated with I. paranaensis, and, although not assessed in this study, could correspond to the vertebrate reservoir of this newly described genospecies in Brazil. Pathogenic roles of "Ca. B. ibitipoquensis" are still unknown. However, its possible vector I. paranaensis is not an anthropophilic tick, so human infections would be unlikely to occur. Our finding enhances the knowledge on Bbsl in South America, highlights the occurrence of ecologically and genetically related genospecies with vastly separated geographical distributions, and calls for the attention to explore a barely known diversity of spirochetes of this group in the region.

Immature ticks on wild birds and the molecular detection of a novel Rickettsia strain in the Ibitipoca State Park, southeastern Brazil.

Birds are recognized hosts of ticks, especially for the immature stages which may harbor various species and strains of Rickettsia. To explore landscapes inhabited by birds and their ticks would expand the knowledge on host-parasite relationships and the rickettsiae. The aim of this paper was to record the diversity of ticks collected on wild birds and assess the phylogenetic position of a novel Rickettsia strain detected in immature ticks. Birds were captured in the Ibitipoca State Park, located in the Minas Gerais state, southeastern Brazil, as part of a long-term research project on the ecology of ticks, birds and Rickettsia. We found three tick species parasitizing birds: Amblyomma aureolatum (63 larvae, 10 nymphs), Haemaphysalis leporispalustris (28 larvae, seven nymphs) and Amblyomma romarioi (27 larvae). Among these, A. aureolatum was the most abundant species including 54% (73/135) of the collected ticks. New tick-host records were: A. romarioi on Turdus amaurochalinus and H. leporispalustris on Thamnophilus caerulescens, Saltator similis and Zonotrichia capensis. Of the 82 ticks tested for Rickettsia spp. by PCR, two larvae (2.5%) of A. romarioi were infected with 'Candidatus Rickettsia paranaensis', a novel putative Rickettsia species closely related to Rickettsia africae, Rickettsia sibirica and Rickettsia parkeri, as corroborated by our phylogenetic analysis. Finally, we present a list of all records of immature stages of H. leporispalustris on passerine birds in Brazil.

Serologic and molecular survey of Rickettsia spp. in dogs, horses and ticks from the Atlantic rainforest of the state of Bahia, Brazil.

Tick-borne spotted fever in Brazil is known to be caused by two agents, Rickettsia rickettsii and Rickettsia parkeri. Nothing was known about these agents in one area of the Atlantic rainforest biome of Bahia state, where during March to June 2016, 356 rural dogs and 69 horses were sampled and their sera were processed through indirect immunofluorescence assay against antigens of R. rickettsii, R. parkeri, Rickettsia amblyommatis and Rickettsia bellii. Ticks collected from these dogs and horses were molecularly tested for the presence of rickettsial DNA. Overall, 16.4% (58/356) dogs and 24.6% (17/69) horses were seroreactive to Rickettsia spp. Five tick species, Rhipicephalus sanguineus sensu lato (s.l.), Amblyomma ovale, A. sculptum, R. microplus, and A. naponense, were collected from dogs, whereas horses were infested by A. sculptum and Dermacentor nitens. A total of 242 ticks from dogs and 62 from horses were analyzed by PCR targeting rickettsiae, which were detected in only 4/27 (14.8%) A. ovale. Fragments of the rickettsial gltA and ompA genes from these four ticks were 100% identical to the Atlantic rainforest strain of R. parkeri. The presence of A. ovale on dogs was positively associated with local canine seroreactivity to R. parkeri. Our results provide evidence for the transmission of R. parkeri strain Atlantic rainforest from A. ovale to domestic dogs within the rural area of Ilhéus, similarly to other areas of the Atlantic rainforest biome of Brazil, where human cases of R. parkeri-caused spotted fever have been reported.

Additional information about tick parasitism in Passeriformes birds in an Atlantic Forest in southeastern Brazil.

The habits of birds make them more or less susceptible to parasitism by certain tick species. Therefore, while some bird species are typically found to be intensely infested, others are relatively unaffected. This study investigated the occurrence of ticks in Passeriformes inhabiting an Atlantic Forest fragment in southeastern Brazil, during the dry and rainy seasons, by means of parasitological indexes and multiple correspondence analysis, to determine the factors that influence tick parasitism in these birds. Data were collected on 2391 ticks, all classified in the Amblyomma genus, from 589 birds. The ticks identified to the species level were A. longirostre, A. nodosum, A. calcaratum, A. parkeri, and A. ovale. Thamnophilidae, Conopophagidae, Thraupidae, Dendrocolaptidae, and Platyrinchidae were the families with the highest prevalence. In terms of parasite intensity, the families Conopophagidae, Thamnophilidae, Thraupidae, Furnariidae, and Pipridae stood out with the highest values. Bird species that are generalists regarding eating habits and habitat occupation tended to have higher parasite loads, as did larger species and those inhabiting the understory. The tick prevalence was higher in the dry season than in the rainy season. The majority of the ticks were collected from the head region, mainly around the eyes and in the nape. Also, this work reports 22 new bird-parasite relations.

Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae), the Cayenne tick: phylogeography and evidence for allopatric speciation.

BACKGROUND: Amblyomma cajennense F. is one of the best known and studied ticks in the New World because of its very wide distribution, its economical importance as pest of domestic ungulates, and its association with a variety of animal and human pathogens. Recent observations, however, have challenged the taxonomic status of this tick and indicated that intraspecific cryptic speciation might be occurring. In the present study, we investigate the evolutionary and demographic history of this tick and examine its genetic structure based on the analyses of three mitochondrial (12SrDNA, d-loop, and COII) and one nuclear (ITS2) genes. Because A. cajennense is characterized by a typical trans-Amazonian distribution, lineage divergence dating is also performed to establish whether genetic diversity can be linked to dated vicariant events which shaped the topology of the Neotropics. RESULTS: Total evidence analyses of the concatenated mtDNA and nuclear + mtDNA datasets resulted in well-resolved and fully congruent reconstructions of the relationships within A. cajennense. The phylogenetic analyses consistently found A. cajennense to be monophyletic and to be separated into six genetic units defined by mutually exclusive haplotype compositions and habitat associations. Also, genetic divergence values showed that these lineages are as distinct from each other as recognized separate species of the same genus. The six clades are deeply split and node dating indicates that they started diverging in the middle-late Miocene. CONCLUSIONS: Behavioral differences and the results of laboratory cross-breeding experiments had already indicated that A. cajennense might be a complex of distinct taxonomic units. The combined and congruent mitochondrial and nuclear genetic evidence from this study reveals that A. cajennense is an assembly of six distinct species which have evolved separately from each other since at least 13.2 million years ago (Mya) in the earliest and 3.3 Mya in the latest lineages. The temporal and spatial diversification modes of the six lineages overlap the phylogeographical history of other organisms with similar extant trans-Amazonian distributions and are consistent with the present prevailing hypothesis that Neotropical diversity often finds its origins in the Miocene, after the Andean uplift changed the topology and consequently the climate and ecology of the Neotropics.

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.

Records and altitudinal assessment of Amblyomma aureolatum and Amblyomma ovale (Acari: Ixodidae) in the State of Rio de Janeiro, southeast Brazil.

Information on the altitudinal distribution of the hard ticks Amblyomma aureolatum and Amblyomma ovale in Brazil is scarce and mainly limited to occasional records. In this study we report our evaluation of records on the altitudinal distribution of A. aureolatum and A. ovale collected from dogs and humans and directly from the environment (host-questing ticks), based on active or passive procedures. The collections were conducted in rural areas of municipalities in the state of Rio de Janeiro, Brazil between 2013 and 2017. Active procedures consisted of dragging or flagging, visual examination of vegetation and removal of ticks present on the authors' clothing or on infested dogs. Overall, 222 ticks were collected. The altitudes at the collection sites ranged from 98 to 1220 m a.s.l. We noted a significant difference in the altitudinal distribution of A. aureolatum and A. ovale (Mann-Whitney U-test, U = 518.5, P < 0.001). The overlap of these two species occurred at altitudes of between 650 and 900 m a.s.l. The results indicated that the higher the altitude, the greater the probability for the occurrence of A. aureolatum and, conversely, the lower the likelihood for the occurrence of A. ovale. The findings of this study improve currrent knowledge on the bioecology of these tick species and have implications for studies on the epidemiology of spotted fever in Brazil.

Molecular Detection of Rickettsia parkeri Strain Atlantic Rainforest in Ticks Parasitizing Small Mammals in Northeastern Brazil.

PURPOSE: Small mammals are important reservoirs of ticks and their pathogens in nature. However, studies reporting these associations are still rare in Brazil. In the present study, we investigated the presence of Rickettsia DNA in ticks parasitizing rodents and marsupials captured in different areas throughout the Atlantic rainforest biome, Bahia, Northeastern (NE), Brazil. METHODS: The study was conducted in five municipalities within of the Atlantic Forest biome, Bahia state, in NE Brazil. Two campaigns were done in each municipality. For host captures Sherman and Tomahawk traps were used, and pitfall traps. After being captured, the hosts were anesthetized and their entire body examined for ticks. When ticks were detected, they were manually removed and stored in eppendorf tubes (1.5 ml) containing absolute PA ethanol for future laboratory analysis (identification of ticks and detection of Rickettsia spp.). RESULTS: A total of 609 mammals were captured. Overall, 208 ticks of the genus Amblyomma and Ixodes were collected: A. ovale, I. loricatus and A. varium. Rickettsia DNA was detected in A. ovale and it was 99-100% of identity to the sequence deposited in GenBank as Rickettsia parkeri strain Atlantica rainforest. CONCLUSION: These results suggest that R. parkeri strain Atlantic rainforest occurs in the region, and A. ovale is likely the vector.

Multi-locus phylogenetic analysis groups the New World bacterium Rickettsia sp. strain ApPR with the Old World species R. africae; proposal of "Candidatus Rickettsia paranaensis".

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".

Seasonal dynamics of ticks (Acari: Ixodidae) on horses in the state of São Paulo, Brazil.

Natural tick infestations were assessed every 14 days on horses over a 2-year period. Amblyomma cajennense adult ticks were counted individually, without detachment from the horses. Larvae and nymphs of A. cajennense were collected using a rubber scraper that scratched engorged immature ticks from the host. Adult females of Anocentor nitens larger than 4mm length were counted on the horses. Blood samples were also obtained from the horses every 14 days and macroclimatic data were obtained for the study period. Infestations of A. cajennense demonstrated distinct peaks of activity for each of the three parasitic stages over each 12-month period, showing a 1-year generation pattern. Larvae predominated from April to July and nymphs from June to October. Adults predominated from October to March with a greater number of adult males than females. Although other studies on seasonal dynamics in the states of Rio de Janeiro and Minas Gerais were performed with the free-living stages of A. cajennense on pastures, the present study in the state of São Paulo, performed with the parasitic stages of A. cajennense on horses, showed similar results to those observed in other states. Infestations by A. nitens demonstrated distinct peaks of activity of adult females (>4 mm), suggesting different tick generations during the year. Infestation with A. nitens was much higher in the first year than the second year which may have been related to horse nutritional status and stocking rate. Although several climatic variables showed statistical significant correlation (r) with tick counts, the determination coefficients (R(2)) were always lower than 0.40, suggesting that any single significant variable (i.e. mean temperature) would not explain the tick distribution pattern over the year. The highest peaks of A. nitens females (>4 mm) were significantly associated with decrease in horse packed cell volumes (R(2)=0.603). The ears and the perineum, tail and groin region accounted for around 70% of all A. nitens females counted on the horses.

Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil.

BACKGROUND: Anaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil. METHODS: We tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum. RESULTS: Among the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum infection in dogs. CONCLUSIONS: This is the first report of A. phagocytophilum in ixodid ticks from Brazil. The detection of A. phagocitophylum in A. cajennense, an aggressive feeder on a wide variety of hosts, including humans, is considered a public health concern.

Detection of Anaplasma phagocytophilum in Brazilian dogs by real-time polymerase chain reaction.

Anaplasma phagocytophilum was detected in dogs from Brazil in the municipalities of Seropédica and Itaguaí, Rio de Janeiro state, by real-time polymerase chain reaction (PCR) using SYBR Green to detect the amplification. Of 253 samples, 18 (7.11%) were positive, with a threshold cycle (Ct) ranging from 31 to 35 cycles. The PCR product from a positive sample was cloned and sequenced. The sequence obtained demonstrated 100% identity with other A. phagocytophilum sequences published in the GenBank database. The analytical sensitivity of RT-PCR using SYBR Green system was able to detect 3 plasmid copies when defined numbers of plasmid copies containing 122 base pairs from the msp2 gene were used. The assay was considered specific when DNA from bacteria (Anaplasma platys, Anaplasma marginale, Ehrlichia canis, Neorickettsia risticii, Rickettsia rickettsii) closely related to A. phagocytophilum was placed in the reaction. These results demonstrate that the canine granulocytic anaplasmosis agent is present in regions in which dogs could be a source of infection for tick vectors. The current study reports the detection of A. phagocytophilum, a zoonotic agent responsible for Human granulocytic anaplasmosis, in Brazilian dogs.

First record of Amblyomma scalpturatum Neumann (Acari: Ixodidae) in the states of Paraná and Roraima, Brazil.

Amblyomma scalpturatum Neumann has been reported in Brazil in the northern (States of Amazonas, Pará and Rondônia) and mid-western regions (States of Mato Grosso and Mato Grosso do Sul). It has been found in association with large and medium sized mammals, as Artiodactyla, Perissodactyla, Xenarthra and Carnivora. In this paper, this tick species is reported for the first time in the Brazilian states of Paraná and Roraima, southern and northern regions, respectively.

Topographical and numerical study of the idiosomal integumentary structures of the larva of four Neotropical species of Amblyomma Koch, 1844 (Acari: Ixodidae).

Integumentary structures of the larvae of Amblyomma longirostre (Koch, 1844), A. parvum Aragão, 1908, A. rotundatum Koch, 1844 and from three populations of A. cajennense (Fabricius, 1787) were studied using light microscopy. A new nomenclature for the localisation of the integumentary structures is proposed. Three types of integumentary structures were identified in the larval idiosoma of the four Amblyomma species: lyrifissures, small glands and large wax glands. These structures were observed isolated or associated over the entire idiosoma, except in the scutum, which lacked lyrifisures and large wax glands. Large wax glands were the most stable within and between the tick species, followed by lyrifissures and small glands. Small glands, although relatively stable, showed the highest number of numerical variations within and between the tick species. Even though there were intra-population variations in the topographical and numerical pattern of some integumentary structures of A. cajennense larvae, there was a definitive pattern for most of the specimens, as showed by the similar modal and mean numbers of integumentary structures per tick side. The patterns of lyrifissures, small glands and large wax glands showed little differences when compared between the four Amblyomma species; however, a few differences were well evident. These differences were sufficient to differentiate larvae of the four species. Thus, we expect that the study of integumentary structures on the larvae of other Amblyomma species will be useful in future taxonomic keys for the identification of Amblyomma larvae from the Neotropical region.

Description of the immature stages and redescription of the female of Ixodes schulzei Aragão & Fonseca, 1951 (Acari: Ixodidae), an endemic tick species of Brazil.

Ixodes schulzei Aragão & Fonseca, 1951 is a tick endemic to Brazil, where nine species of Ixodes Latreille, 1796 are currently known to occur. Larvae, nymphs and females of I. schulzei were obtained from a laboratory colony originating from an engorged female collected on a free-living water rat Nectomys squamipes from the Santa Branca municipality, São Paulo State. Only female ticks were obtained from engorged nymphs. Unfed immature and female adult specimens were measured and the descriptions were based on optical and scanning electron microscopy, as were drawings of some features of the larva. Both immature stages present the very long palpi and basis capituli, and the female has large, contiguous porose areas. However, the basis capituli is triangular, with a slight central elevation in the larva and nymph, whereas in the female this area is depressed. The I. schulzei types deposited at the FIOCRUZ (Instituto Oswaldo Cruz) were also examined, as was other material from collections, such as the IBSP (Coleção Acarológica do Instituto Butantan), CNC-FMVZ/USP (Coleção Nacional de Carrapatos da Faculdade de Medicina Veterinária e Zootecnia da USP) and USNTC (United States National Tick Collection). In addition, the relationship between I. schulzei and other immature neotropical species of Ixodes is discussed.

The role of ear mites of the genus Raillietia (Acari: Raillietiidae) in otitis of domestic ruminants / O papel dos ácaros do gênero Raillietia (Acari: Raillietiidae) na otite de ruminantes domésticos

This paper presents a hypothesis on the involvement of species of the genus Raillietia Trouessart in the pathology of parasitic otitis in domestic ruminants. The chelicerae and claws of the tarsi are structures that contribute significantly towards producing primary lesions. The movable digits of the chelicerae accidentally injure the epithelium that lines the ear canal, while the claws of the tarsi perforate the epithelium, allowing the mites to affix themselves while feeding. The lesions in the epithelium caused by the chelicerae and the claws favor multiplication of the bacteria that inhabit the ear canal, resulting in typical cases of otitis.

Neste trabalho apresentamos uma hipótese para explicar o envolvimento dos ácaros do gênero Raillietia na patogênese da otite parasitária em ruminantes domésticos. As quelíceras e unhas dos tarsos são estruturas que contribuem significativamente para produzir as lesões primárias no epitélio. Os dígitos móveis das quelíceras acidentalmente causam injúria no epitélio que reveste o canal auditivo, enquanto as unhas dos tarsos perfuram o epitélio para permitir a fixação dos ácaros durante sua alimentação. As lesões no epitélio causadas pelas quelíceras e unhas dos tarsos favorecem a multiplicação das bactérias que habitam o canal auditivo, resultando em casos típicos de otite.