Ticks (Acari: Ixodidae) and rickettsiae associated with wild boars in a rural area of Minas Gerais, Brazil.

Wild boars or feral pigs are classified by the Brazilian Institute for the Environment and Renewable Resources (IBAMA) in "Category I of invasive exotic species". They cause economic losses, harm the environment, serve as hosts and reservoirs for several zoonotic disease agents, and provide a blood meal for tick species that act as vectors for zoonotic diseases. The objective of this study was to identify tick species on wild boars, assess host-seeking ticks in the related environment, and identify other potential tick hosts coexisting with wild boars on a farm located in the state of Minas Gerais, southeastern Brazil. Additionally, the study aimed to determine the presence of rickettsiae in these arthropods and assess the exposure of wild boars to rickettsiae species from the Spotted Fever Group and Rickettsia bellii through serology. A total of 3585 host-seeking ticks from three species (Amblyomma sculptum - 41.58%; Amblyomma dubitatum - 0.39% and Rhipicephalus microplus - 0.05%) were collected in the environment and A. sculptum was the most abundant species. Thirty-one wild boars were evaluated, resulting in the collection of 415 ticks, all of which were A. sculptum. Rickettsia DNA was not detected in samples of A. sculptum and R. microplus from the environment or in A. sculptum ticks from wild boars. However, all A. dubitatum ticks (n = 14) had Rickettsia bellii DNA confirmed by the species-specific PCR protocol. Out of the 31 serum samples from wild boars, 24 reacted with at least one Rickettsia antigen. Among these, seven individuals exhibited a reaction to a probable homologous antigen (PHA) of three rickettsiae species: R. rickettsii (n = 3), R. amblyommatis (n = 3) and R. rhipicephali (n = 1). Despite the high prevalence of seroreactivity, titers were low, indicating limited exposure to Rickettsia spp. Camera traps generated 874 animal records, capturing a total of 1688 individuals. At least 11 species of birds and 14 species of mammals (12 wild and two domestic) shared the environment with wild boars and potentially shared ticks with them. These findings provide baseline information for understanding the sharing of ticks and tick-borne pathogens between wild boars and other animals within the Cerrado biome. Further studies are necessary to monitor the potential and actual risk of wild boars to harbor infected ticks and their role in the transmission and maintenance cycle of Rickettsia spp.

Density and behavior of capybara (Hydrochoerus hydrochaeris) ticks (Acari: Ixodidae) Amblyomma sculptum and Amblyomma dubitatum with notes on Rickettsia bellii infection: Assessing human exposure risk.

In several urban and peri­urban areas of Brazil, populations of Amblyomma sculptum and Amblyomma dubitatum ticks are maintained by capybaras (Hydrochoerus hydrochaeris). In some of these areas, this host and these tick species are associated with Brazilian spotted fever (BSF), a lethal human disease caused by the bacterium Rickettsia rickettsii. In this work, we evaluated the risk of human exposure to these tick species using four collection techniques to discern host-seeking behavior. The study was carried out in 10 urban sites inhabited by capybaras in Uberlândia, a BSF-free municipality in southeastern Brazil. Ticks were collected in areas of 400 m2 at each site and at three seasons. Within the same municipality, the distance and speed of A. sculptum nymphs moving towards the CO2 traps were evaluated. In a sample of ticks Rickettsia DNA was investigated. During the study period, 52,953 ticks were collected. Among these, 83.4 % were A. sculptum (1,523 adults, 10,545 nymphs and 32,104 larvae) and 16.6 % were A. dubitatum (464 adults, 2,153 nymphs and 6,164 larvae). An average annual questing tick density of 4.4/m² was observed, with the highest density recorded at one site in autumn (31.8/m²) and the lowest in summer at another site (0.03/m²). The visual search yielded the highest proportion of A. sculptum larvae, constituting 47 % of the total and 63.6 % of all A. sculptum larvae. In contrast, CO2 traps collected a greater proportion of nymphs and adults of A. sculptum ticks. In the case of A. dubitatum, the CO2 trap was the most efficient technique with 57.7 % of captures of this species, especially of nymphs (94.5 % of captures) and adults (97.8 % of captures). Ticks' ambush height on vegetation (9 to 77 cm), observed by visual search 30 times, yielded a total of 20,771 ticks. Of these, 28 (93 %) were A. sculptum ticks, with only two (7 %) identified as A. dubitatum ticks. Among A. sculptum ticks, the nymph was the most attracted stage to humans and larva in the case of A. dubitatum. Amblyomma sculptum adults and nymphs were significantly more attracted to humans than those of A. dubitatum, but A. dubitatum larvae were significantly more attracted than the same stage of A. sculptum. The maximum distance and speed of horizontal displacement for A. sculptum nymphs were five meters and 2.0 m/h, respectively. The only species of Rickettsia detected in ticks, exclusively in A. dubitatum, was R. bellii. Importantly, it was observed that the higher the proportion of A. sculptum in the community of ticks, the lower the rate of infection of A. dubitatum by R. bellii. In conclusion, host-seeking behavior differed between the two tick species, as well as between stages of the same species. A greater restriction of A. dubitatum ticks to the soil was observed, while larvae and nymphs of A. sculptum dispersed higher in the vegetation. The behavior presented by A. sculptum provides greater opportunities for contact with the hosts, while A. dubitatum depends more on an active search for a host, the hunter behavior. Taken together, these observations show that a human being crossing an area infested with A. sculptum and A. dubitatum ticks will have almost exclusive contact with A. sculptum larvae and/or nymphs. Humans in a stationary position (sitting, lying or immobile) are exposed to both tick species, but they are more attractive to adults and mainly nymphs of A. sculptum compared to the corresponding stages of the tick A. dubitatum. The negative effect of A. sculptum on A. dubitatum infection by R. bellii deserves further studies.

Solid epithelioid peritoneal mesothelioma with pulmonary metastasis in feline.

Mesothelioma is a rare malignant neoplasm that affects the mesothelial cells lining the thoracic and abdominal cavities, such as the pleura, peritoneum, and pericardium. It is most prevalent in dogs and cattle, but the causes of this disease in animals are uncertain. In felines, it mainly affects the pleura, with an unfavorable prognosis. This paper explores a rare case of metastatic peritoneal mesothelioma in a 2-year-old female mixed breed cat, emphasizing its uniqueness due to the feline's age. The patient, previously treated at a private clinic, presented moderate abdominal distension as the only clinical sign. Abdominal ultrasound and peritoneal fluid cytology led to the provisional diagnosis of mesothelioma/carcinomatosis. One day after exploratory laparotomy, the animal died and was subsequently sent for necropsy. During macroscopic analysis, nodules were observed in the peritoneum, diaphragm, omentum, stomach serosa, and large intestine, and the diagnosis of solid epithelioid peritoneal mesothelioma with lung metastasis was confirmed after microscopic analysis. The diagnosis of mesothelioma is challenging, and the importance of immunohistochemical panels with specific markers such as cytokeratin AE1/AE3 and calretinin is highlighted. Considering that mesothelioma is a pathology with a poor prognosis, it is essential to include this disease in the list of differential diagnoses within veterinary oncology.

O mesotelioma é uma neoplasia maligna rara que afeta as células mesoteliais que revestem as cavidades torácica e abdominal, como a pleura, o peritônio e o pericárdio. É mais prevalente em cães e bovinos, mas as causas desta doença em animais são incertas. Nos felinos acomete principalmente a pleura, com prognóstico desfavorável. Este artigo explora um caso raro de mesotelioma peritoneal metastático em uma gata sem raça definida de 2 anos de idade, enfatizando sua singularidade devido à idade do felino. O paciente, previamente atendido em clínica particular, apresentava distensão abdominal moderado como único sinal clínico. A ultrassonografia abdominal e a citologia do líquido peritoneal levaram ao diagnóstico provisório de mesotelioma/carcinomatose. Um dia após a laparotomia exploratória, o animal veio a óbito e posteriormente encaminhado para necropsia. Durante a análise macroscópica, foram observados nódulos no peritônio, diafragma, omento, serosa estomacal e intestino grosso e o diagnóstico de mesotelioma peritoneal epitelioide sólido com metástase pulmonar foi confirmado após análise microscópica. O diagnóstico do mesotelioma é desafiador, sendo destacada a importância de painéis imunohistoquímicos com marcadores específicos como citoqueratina AE1/AE3 e calretinina. Considerando que o mesotelioma é uma patologia de prognóstico ruim, é fundamental incluir esta doença na lista de diagnósticos diferenciais dentro da oncologia veterinária.

Survey of Rickettsia spp. in ticks (Acari: Ixodidae) infesting opossums (Didelphis albiventris) and capybaras (Hydrochoerus hydrochaeris) from an urban park in southeastern Brazil.

Ticks are hematophagous arthropods and, during feeding, may transmit pathogens to vertebrate hosts, including humans. This study aimed to investigate the presence of Rickettsia spp. in ticks collected between 2010 and 2013 from free-ranging capybaras (Hydrochoerus hydrochaeris) and opossums (Didelphis albiventris) that inhabit Sabiá Park in Uberlândia, Brazil. Overall, 1,860 ticks were collected: 1,272 (68.4%) from capybaras (487 of the species Amblyomma sculptum, 475 adults and 12 nymphs; 778 Amblyomma dubitatum, 727 adults and 51 nymphs; and seven larva clusters of the genus Amblyomma); and 588 (31.6%) from opossums (21 A. sculptum, one adult and 20 nymphs; 79 A. dubitatum, all nymphs; 15 Ixodes loricatus, 12 adults and three nymphs; 457 Amblyomma sp. larva clusters; 15 Ixodes sp. larva clusters; and one Argasidae larva cluster). Out of 201 DNA samples tested for the presence of Rickettsia spp. DNA using polymerase chain reaction (PCR) 12 showed amplification of a gtlA gene segment that was specific to Rickettsia bellii, a bacterium non-pathogenic to humans. As there has been a report showing serological evidence of infections caused by Rickettsia species of the spotted fever group (SFG) in capybaras and opossums in the park, including Rickettsia rickettsii, the etiological agent of Brazilian spotted fever, and considering the presence of A. sculptum ticks, which are aggressive to humans, as well as these vertebrate hosts, which are amplifiers of R. rickettsii, it is important to monitor the presence of SFG rickettsiae in the Sabiá Park, which is visited daily by thousands of people.

Molecular detection of Borrelia sp. in Ornithodoros cavernicolous (Acari: Argasidae) in midwestern Brazil.

Ticks are obligate hematophagous parasites that can transmit to vertebrate hosts several pathogens, including viruses, bacteria, protozoa and helminths. Among these agents, some Borrelia species some Borrelia species cause disease in humans and other vertebrate hosts; therefore, they have medical and veterinary health importance. To gather additional information on Borrelia species in Brazil, the current study aimed to detect the presence of these species in Ornithodoros cavernicolous ticks collected in September 2019 from cement pipes that are used by bats as shelter in a farm located in the midwestern region of Brazil. DNA samples obtained from 18 specimens of O. cavernicolous were subjected of two polymerase chain reactions, targeting a segment of the Borrelia fla B gene. Of the samples tested, only one (6 %, 1/18) showed amplification. The nucleotide sequence of the amplified DNA showed more than 97 % (293/300) identity with a sequence of a Borrelia sp. detected in blood collected from a bat from Macaregua Cave, Colombia, and more than 97 % (292/300) detected in lungs from vampire bats from northeastern Brazil. The deduced amino acid sequences were identical to each other. Phylogenetic analysis indicated that these sequences formed a group of Borrelia species (putatively associated with bats) that is closely related to sequences of Borrelia species of the Lyme borreliosis group. Further investigations should be carried out in order to determine whether the sequence of the Borrelia sp. we found belongs to a new taxon. It will also be of great importance to determine which vertebrate hosts, besides bats, O. cavernicolous ticks can parasitize in order to investigate whether the Borrelia sp. we found may be transmitted and cause disease to the other vertebrate hosts.

Low Genetic Diversity of the Only Clade of the Tick Rhipicephalus microplus in the Neotropics.

This study addresses the variability of the mitochondrial cytochrome oxidase subunit I (COI) and 16S rDNA (16S), and nuclear internal transcriber spacer ITS2 (ITS2) genes in a set of field-collected samples of the cattle tick, Rhipicephalus microplus (Canestrini, 1888), and in geo-referenced sequences obtained from GenBank. Since the tick is currently considered to be a complex of cryptic taxa in several regions of the world, the main aims of the study are (i) to provide evidence of the clades of the tick present in the Neotropics, (ii) to explore if there is an effect of climate traits on the divergence rates of the target genes, and (iii) to check for a relationship between geographical and genetic distance among populations (the closest, the most similar, meaning for slow spread). We included published sequences of Rhipicephalus annulatus (Nearctic, Afrotropical, and Mediterranean) and R. microplus (Afrotropical, Indomalayan) to fully characterize the Neotropical populations (total: 74 16S, 44 COI, and 49 ITS2 sequences included in the analysis). Only the clade A of R. microplus spread in the Nearctic-Neotropics. Both the K and Lambda's statistics, two measures of phylogenetic signal, support low divergence rates of the tested genes in populations of R. microplus in the Neotropics. These tests demonstrate that genetic diversity of the continental populations does not correlate either with the geographic distance among samples or with environmental variables. The low variability of these genes may be due to a combination of factors like (i) the recent introduction of the tick in the Neotropics, (ii) a large, effective, and fast exchange of populations, and (iii) a low effect of climate on the evolution rates of the target genes. These results have implications for the ecological studies and control of cattle tick infestations.

Perception of Ticks and Tick-Borne Diseases Worldwide.

In this comprehensive review study, we addressed the challenge posed by ticks and tick-borne diseases (TBDs) with growing incidence affecting human and animal health worldwide. Data and perspectives were collected from different countries and regions worldwide, including America, Europe, Africa, Asia, and Oceania. The results updated the current situation with ticks and TBD and how it is perceived by society with information bias and gaps. The study reinforces the importance of multidisciplinary and international collaborations to advance in the surveillance, communication and proposed future directions to address these challenges.

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.

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.

Use of selected samples to diagnose a tricky feline viral disease in a cat with uveitis and neurological signs.

This case involved a 2-year-old neutered male domestic mixed-breed cat that was rescued from the street eight months earlier. The animal presented with weakness, hyporexia, progressive weight loss, fatigue, uveitis, pale mucous membranes, dehydration (7%), and pelvic limb paresis. Aqueous humor was collected for molecular analysis for the differential diagnosis of potential etiological agents [Feline coronavirus (FCoV), Feline leukemia virus (FeLV), Feline immunodeficiency virus (FIV), Toxoplasma gondii, Cryptococcus spp., Felid herpesvirus-1 (FHV-1) and Bartonella spp.] of feline uveitis. The sample was positive by real-time reverse transcription-polymerase chain reaction (RT-qPCR) for FCoV and RT-qPCR and real-time polymerase chain reaction (qPCR) for FeLV and qPCR FIV. The cat was euthanized due to poor clinical outcomes and prognosis. A cerebrospinal fluid (CSF) sample was collected and tested, and the same pathogens were found in the aqueous humor. Small-cell follicular multicenter lymphoma and multifocal pyogranulomatous meningoencephalitis were observed upon histopathological analysis. In this study, aqueous humor and cerebrospinal fluid samples were efficient for the detection of coinfection with FIV, FeLV, and FCoV.

O caso refere-se a um gato de dois anos de idade, sem raça definida, resgatado da rua há oito meses. O animal apresentava fraqueza, hiporexia, emagrecimento progressivo, cansaço fácil, uveíte, mucosas pálidas, desidratação (7%) e paresia de membros pélvicos. O humor aquoso foi coletado para o diagnóstico molecular diferencial de potenciais agentes etiológicos [coronavírus felino (FCoV), vírus da leucemia felina (FeLV), vírus da imunodeficiência felina (FIV), Toxoplasma gondii, Cryptococcus spp., herpesvírus felino tipo 1 (FHV-1) and Bartonella spp.] causadores de uveíte felina. A amostra foi positiva na reação em cadeia da polimerase precedida por transcrição reversa em tempo real (RT-qPCR) para FCoV, RT-qPCR e reação em cadeia da polimerase em tempo real (qPCR) para FeLV e qPCR para FIV. O animal foi submetido à eutanásia - devido ao quadro clínico e prognóstico desfavorável. Amostra de líquido cefalorraquidiano (LCR) foi coletada e testada, confirmando a identificação dos mesmos patógenos encontrados no humor aquoso. Linfoma multicêntrico folicular de pequenas células e meningoencefalite piogranulomatosa multifocal foram observados na análise histopatológica. Neste relato, as amostras de humor aquoso e líquido cefalorraquidiano foram eficientes para a detecção de coinfecção por FIV, FeLV e FCoV.

Evaluation of a mimotope of the Rickettsia outer membrane protein A (OmpA) as an antigen in enzyme-linked immunosorbent assay to detect rickettsiosis in capybaras (Hydrochoerus hydrochaeris), horses (Equus caballus), and opossums (Didelphis sp.).

Rickettsia rickettsii is the etiological agent of Rocky Mountain spotted fever, which is an important tick-borne zoonosis and, in Brazil, it causes Brazilian spotted fever, which has high lethality rate. This study aimed to evaluate a synthetic peptide corresponding to a segment of the outer membrane protein A (OmpA) as an antigen in a serological test for the diagnosis of rickettsial infections. The amino acid sequence of the peptide was selected by predicting B cell epitopes using B Cell Epitope Prediction (Immune Epitope Database and Analysis Resource) and Epitopia and OmpA sequences of Rickettsia rickettsii strain 'Brazil' and Rickettsia parkeri strains 'Maculatum 20' and 'Portsmouth'. A peptide with amino acid sequence common to both Rickettsia species was synthesized and arbitrarily named OmpA-pLMC. To evaluate this peptide in enzyme-linked immunosorbent assay (ELISA), serum samples of capybara (Hydrochoerus hydrochaeris), horse (Equus caballus), and opossum (Didelphis albiventris) that had been previously tested by indirect immunofluorescence assay (IFA) for rickettsial infection were separated into IFA-positive and IFA-negative groups and used in the assay. There were no significant differences in ELISA optical density (OD) values between IFA-positive and IFA-negative groups with horse samples. The mean OD values were significantly higher in the IFA-positive capybara serum samples (IFA-pos vs. IFA-neg = 2.389 ± 0.761 vs. 1.760 ± 0.840). However, receiver operating characteristic (ROC) curve analysis did not show significant diagnostic parameters. On the other hand, 12 out of 14 (85.7%) opossum samples of the IFA-positive group showed reactivity in ELISA, and this was significantly higher than of the IFA-negative group (0.7196 ± 0.440 vs. 0.2318 ± 0.098, respectively; 85.7% sensitivity, 100% specificity). Therefore, our results show that OmpA-pLMC has a potential to be used in immunodiagnostic assays to detect spotted fever group rickettsial infections.

The inoculation eschar of Rickettsia parkeri rickettsiosis in Brazil: Importance and cautions.

Two well characterized tick-borne rickettsioses occur in Brazil. Rickettsia rickettsii caused spotted-fever, transmitted by Amblyomma sculptum and Amblyomma aureolatum, is a severe disease with a high case-fatality rate in the southeastern region of the country. Rickettsia parkeri strain Atlantic rainforest infections transmitted by adult Amblyomma ovale ticks cause a milder non-lethal febrile disease with an eschar (necrosis) at the tick bite site. Clinical diagnosis of rickettsiosis is challenging, particularly during the early stages of the illness when signs and symptoms are non-specific. Since eschar at the tick bite site has emerged as the main clinical feature of mild R. parkeri infections and used to differentiate it from severe R. rickettsii infection, its proper recognition, distinction from other tick bite lesions, and boundaries as a clinical tool must be highlighted. Of importance, eschars induced by Rickettsia must be differentiated from dermatoses caused by other tick-borne skin infections as well from lesions caused by the tick bite itself. We herein highlight information on eschar in rickettsial diseases in Brazil and discuss the need for further research on its clinical relevance and application in the diagnosis of spotted fever caused by R. parkeri strain Atlantic rainforest. In particular, we draw attention to diagnosis of other febrile diseases in the presence of concomitant tick bites.

One Health Approach to Identify Research Needs on Rhipicephalus microplus Ticks in the Americas.

We aim to provide a harmonized view of the factors that affect the survival and promote the spread of R. microplus in the Neotropics, approaching its different facets of biology, ecology, distribution, and control. We review the interactions among environmental niche, landscape fragmentation, vegetal coverage (abiotic traits), and the biotic aspects of its ecology (abundance of domesticated or wild competent hosts), proposing emerging areas of research. We emphasize a holistic view integrating an economically and ecologically sustainable control of infestations and transmitted pathogens by R. microplus in the Neotropics. Examples of research link the trends of climate, the composition of the community of hosts, the landscape features, and a tailored management based on ecological grounds. Our view is that factors driving the spread of R. microplus are complex and deeply interrelated, something that has been seldom considered in control strategies. The effects of climate may affect the dynamics of wildlife or the landscape composition, promoting new patterns of seasonal activity of the tick, or its spread into currently free areas. In this paper we encourage a One Health approach highlighting the main aspects governing the components of the tick's life cycle and its interactions with livestock and wild animals.

Seasonal dynamics of Amblyomma sculptum: a review.

BACKGROUND: Amblyomma sculptum is a hard tick that is associated with domestic animals and the transmission of Brazilian spotted fever. This association has motivated several field studies on this ixodid tick within its distribution area in South America. Thorough knowledge of the seasonal dynamics of A. sculptum in different ecological scenarios is required in order to better understand the biological characteristics of this tick and develop techniques for the control and prevention of diseases transmitted by this vector. In this article, we systematically review the seasonal dynamics of A. sculptum and tick collection methodology. METHODS: A systematic search of the Scopus, Web of Science, PubMed and Scielo databases was carried out for articles (including dissertations and theses) on the population dynamics of A. sculptum. The inclusion criterion was the report of seasonal dynamic studies on A. sculptum through surveys carried out for at least 1 year with, as methodology, tick collection in the environment and/or tick count/collection on A. sculptum primary hosts (horses or capybaras). Studies carried out before the reclassification of Amblyomma cajennense sensu lato in 2014, which referred to Amblyomma cajennense in areas where it is currently known that only A. sculptum occurs, were also included. Articles meeting the inclusion criterion, but not available in online databases, were also added based on the authors' experience on the subject. Sixteen articles and one thesis were selected for inclusion in this systematic review. RESULTS: Most of the studies were carried out in the southeastern region of Brazil, with a few also carried out in the northeast, center-west and south of Brazil and northwest of Argentina. Five techniques/methods were applied across these studies: CO2 traps, dragging, flagging, visual searches and tick counting on animals, used alone or in combination. CONCLUSION: Seasonal dynamics of A. sculptum was found to be similar in almost all of the areas studied, with larvae predominating during the autumn, nymphs in the winter and adults in the spring and summer.

Immunophenotypical and pathological changes in dogs experimentally infected with Ehrlichia canis.

Canine monocytic ehrlichiosis (CME) is one of the most important tick-borne diseases worldwide, with multisystemic presentations. Immune dysregulation has been proposed as the primary mechanism involved in its pathogenesis and in tissue injury in dogs with CME. Experimental infection of German Shepherd dogs in the present study demonstrated that CME caused marked pathological changes in their lymph nodes and spleen, and also gave rise to mononuclear infiltration in organs and tissues. Immunophenotyping of cells in lymph nodes, spleen and injured tissues highlighted differences in lymphocyte subsets, local expression of immunoglobulin subclasses and MHCII molecules between infected and control dogs. These findings suggest that the immunophenotypic and immunopathological changes in dogs with acute experimental CME are related to Th1 bias and compartmentalized immune response.

Comparing scapular morphology of Amblyomma sculptum and Amblyomma dubitatum nymphs allows a fast and practical differential diagnosis of ticks in highly infested areas with dominance of these two species.

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.

Novel genotypes of Hepatozoon spp. in small mammals, Brazil.

BACKGROUND: Small mammals (rodents and marsupials) have been poorly explored for the occurrence of apicomplexan (genus Hepatozoon and genera of the order Piroplasmorida) and Anaplasmataceae agents in Brazil. Thus, this study investigated the occurrence of Hepatozoon spp., Piroplasmorida, and Anaplasmataceae agents in small mammals in seven forest fragments in Brazil. METHODS: During 2015-2018, small mammals were captured in six forest fragments in the State of São Paulo (Cerrado and Atlantic Forest biomes) and one fragment in the State of Mato Grosso do Sul (Pantanal biome). Mammal blood, liver, spleen, and lung samples were tested molecularly for the presence of DNA of Hepatozoon, Piroplasmorida, and Anaplasmataceae agents. RESULTS: A total of 524 mammals were captured, comprising seven species of marsupials, 14 rodents, two carnivores, and one Cingulata. Four novel haplotypes (1, 2, 3, 4) of Hepatozoon spp. were detected in small mammals from different biomes. In São Paulo state, haplotype 1 was detected in rodents from Cerrado and a transition area of Cerrado and Atlantic Forest biomes, whereas haplotype 2 was detected in rodents from the Atlantic Forest biome. On the other hand, haplotypes 3 and 4 were restricted to rodents and marsupials, respectively, from the Pantanal biome of Mato Grosso do Sul. No host species shared more than one haplotype. Despite these distinct geographical and host associations, our phylogenetic analyses indicated that the four Hepatozoon haplotypes belonged to the same clade that contained nearly all haplotypes previously reported on rodents and marsupials, in addition to several reptile-associated haplotypes from different parts of the world. No mammal samples yielded detectable DNA of Piroplasmorida agents. On the other hand, the Anaplasmataceae-targeted polymerase chain reaction (PCR) assay amplified a sequence 100% identical to the Wolbachia pipientis endosymbiont of the rodent filarid Litomosoides galizai. CONCLUSIONS: We report a variety of Hepatozoon haplotypes associated with small mammals in three Brazilian biomes: Cerrado, Atlantic Forest, and Pantanal. Through phylogenetic analyses, the Hepatozoon agents grouped in the rodent-marsupial-reptile large clade of Hepatozoon spp. from the world. The detection of a W. pipientis associated with the rodent filarid L. galizai indicates that the rodent was infected by filarial nematodes.

Ticks on humans in an Atlantic rainforest preserved ecosystem in Brazil: Species, life stages, attachment sites, and temporal pattern of infestation.

We herein describe exposure and tick attachment to researchers and field staff within the greatest preserved inland Atlantic rainforest, located in Brazil, over a non-consecutive period of five years. Overall, 433 ticks from five species were collected, Amblyomma coelebs (n = 281), Amblyomma brasiliense (n = 79), Amblyomma incisum (n = 43), Amblyomma ovale (n = 4), Haemaphysalis juxtakochi (n = 4). Clusters of larvae belonged to the genus Amblyomma (n = 21) and Haemaphysalis (n = 1). Both Haemaphysalis and Amblyomma species were observed crawling on humans but only ticks from the latter genus were found attached. Most recovered ticks (59%) were found attached and, whereas A. brasiliense was the main host-seeking tick species in the forest, A. coelebs nymphs were the ones that overwhelmingly attached to humans (70% of all attachments). At the same time only 0.4% of human bites were of A. ovale, the Rickettsia parkeri strain Atlantic rainforest vector tick in Brazil. Tick attachment sites were recorded for 132 ticks and lower/upper limbs and the trunk were more parasitized than the head/neck. Tick bites were recorded throughout the year; while larvae did not show an evident seasonal pattern, nymphs attached to humans mainly in autumn and winter and the few adult bites were recorded in spring. Our results highlighted A. coelebs nymphs as major human-biters and evidenced the necessity of studies that discern human tick bites that occur after exposure to host-seeking ticks from those that occur after tick displacement from hosts that carry ticks to human dwellings.

Ticks (Acari: Ixodidae) on marsh deer (Blastocerus dichotomus) at a conservation center: infestation and Rickettsia parkeri infection dynamics along nine years.

The objective of this work was to evaluate the relationship between the populations of the ticks Amblyomma triste Koch, 1844, Rhipicephalus microplus (Canestrini, 1888), Amblyomma sculptum Berlese, 1888, the pathogenic bacteria Rickettsia parkeri and a marsh deer (Blastocerus dichotomus) population after its removal from a pristine environment. For this purpose, ticks were collected from the cervical region of deer at the Marsh Deer Conservation Center in Promissão, São Paulo State, Brazil for nine consecutive years (2000 - 2008). Deer in captivity at the Center were kept in 2,000 m² paddocks surrounded by two-meter-high fences in the Tiete-river marsh. In total, 1,012 ticks of 26 deer were collected. Prevalence of the species A. triste among tick-infested hosts was the highest in the first triennium but decreased to the second and further to the third triennium. In contrast, the R. microplus prevalence amidst infested host population, increased from the first to the third triennium and was the species that attained the highest infestation intensity. Amblyomma sculptum was the tick with the lowest infestation prevalence and intensity throughout the period. The change in the proportion between the two most prevalent species was attributed to the new environment, specifically its restricted size and within it a dry area more suitable for R. microplus. DNA of 424 ticks processed in 276 pools was tested for Rickettsia genetic material by polymerase chain reactions (PCR). Twenty samples of the study were positive for the rickettsial gltA gene. Of these, 18 were from A. triste ticks and revealed the presence of the ompA spotted fever group gene as well. Eleven samples were sequenced and showed 100% identity with R. parkeri sensu stricto. Two samples from R. microplus did not amplify ompA gene neither yielded product in a PCR specific for Rickettsia bellii. Sequencing of the gltA gene in the DNA of these two ticks was also 100% identical with R. parkeri s.s. In conclusion, the changes in the deer environment modified the tick populations but maintained, at least temporary, R. parkeri bacteria in A. triste ticks. Rhipicephalus microplus was refractory to the R. parkeri infection and bacterial DNA in this tick species indicated DNA spill over from other tick species. It was demonstrated that captive marsh deer may sustain cattle tick populations on its own. This would hardly occur under pristine conditions because of the inadequacy the marsh deer´s naturally humid habitat for the cattle tick. However, deer transportations of R. microplus ticks from one farm to another may occur whenever habitat loss pushes this wild animal towards farms.

Small mammals, ticks and rickettsiae in natural and human-modified landscapes: Diversity and occurrence of Brazilian spotted fever in Brazil.

We studied communities of small mammals and their ticks in endemic (E) and non-endemic (NE) areas for Brazilian spotted fever (BSF), aiming to infer if diversity parameters of parasites and hosts could be related to occurrence and prevalence of rickettsial infection, especially Rickettsia rickettsii. We compared E and NE areas in human-modified landscapes (HMLs) and natural areas (BIO) with no report of BSF cases. Composition and equitability were important components of diversity explaining differences among areas. The marsupial Didelphis albiventris was dominant in HMLs, but not in natural areas, and this opossum was the main host for the tick Amblyomma sculptum, principal vector of R. rickettsii, especially in E areas. Communities of ticks were dominated by A. sculptum, followed by Amblyomma dubitatum in E areas. In NE areas, this dominance was inverted, with more A. dubitatum than A. sculptum infesting small mammals, but the numbers of ticks were much lower than in E areas. Composition and abundance of ticks in natural areas were very dissimilar from HMLs, with the lowest tick burdens. Didelphis albiventris in E areas presented higher seroprevalence and endpoint titres against R. rickettsii than in other areas. At least three Rickettsia species, non-pathogenic to humans, were detected in natural areas (Rickettsia bellii, Rickettsia amblyommatis and 'Candidatus Rickettsia andeanae'), and only one non-pathogenic species in HMLs (R. bellii). Our results suggest that higher diversity of ticks, hosts and rickettsiae could be relevant factors in buffering the effect in BSF occurrence. Particularly for D. albiventris, its importance has to be quantified in further studies considering the epidemiological scenario of BSF.