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Abstract Background In 2022, fluralaner was launched on the market for use in the control of the cattle tick Rhipicephalus microplus after showing 100% efficacy in registration trials against the causative agents of cattle tick fever (TFAs). The aim of the present study was to determine whether a strategic control regimen against R. microplus using fluralaner (FLU) in Holstein calves grazing in a tropical region would alter the enzootic stability status of cattle tick fever, triggering outbreaks in these animals up to 22 months age. Methods In this study, a group of calves treated with FLU was compared with a control group treated with the regimen currently being used on the farm, which consisted of the fipronil + fluazuron formulation (FIFLUA). In the first experiment, the efficacy of the FIFLUA pour-on formulation was evaluated in a field study. In the second experiment, which lasted 550 days, two experimental groups (n = 30/group) of Holstein calves naturally infested with R. microplus were analyzed. Calves aged 4 to 10 months received either a specific treatment regimen with FLU (experimental group) or FIFLUA (control group). During this period, tick counts, animal weight measurement, feces collection (to determine eggs and oocysts per gram of feces), tick fever monitoring, blood smears (to ascertain enzootic stability of the herd), PCR testing for TFAs and serology (indirect enzyme-linked immunosorbent assay [iELISA]) were performed. All calves were evaluated for signs of tick fever between ages 11 and 22 months. Results FIFLUA showed an acaricidal efficacy of > 90% from post-treatment days 14 to 35. Regarding treatments against the TFAs, the average number of treatments was similar between groups, but animals treated with FLU had a smaller reduction in packed cell volume on some of the evaluation dates of the second and third treatment against TFAs. In calves aged 10 months in the FLU group, B. bovis was not detected by PCR (0/15 samples), 40% of the samples had antibody titers and 33% (10/30) of the samples had positive blood smears. Regarding B. bigemina, > 86% of the samples in both groups tested positive for B. bigemina DNA and antibodies; there was no difference in the antibody titers between the groups. There were no clinical cases of cattle tick fever in calves aged 11 to 22 months. Conclusions In comparison with the control treatment, the strategic control regimen against R. microplus with FLU that was implemented in the present study did not negatively affect the enzootic stability status of A. marginale and B. bigemina in the herd up to 22 months of age. The enzootic stability status of B. bovis was not reached by either group. These results likely represent a characteristic of the local tick population, so further studies should be performed. Graphical Abstract

Wild boars (Sus scrofa L.) are considered among the most harmful invasive species worldwide, causing irreversible ecosystem damage, acting as zoonotic spreaders and reservoirs, threatening human and animal health, and having an important economic impact. Accordingly, the present study has assessed the rickettsial exposure, tick infestation of wild boars, and rickettsial DNA presence in ticks from infested animals from the Cerrado biome in midwestern Brazil. Anti-Rickettsia spp. antibodies were detected in serum samples of wild boars by immunofluorescence assay. Overall, 106/285 (37.2%) wild boar serum samples from 13 to 18 (72.2%) municipalities showed seroreactivity to at least one of the four Rickettsia spp. antigens tested, the largest number of wild boars serologically tested to Rickettsia spp. in this type of study. Among the 106 seroreactive animals, 34 showed possible homologous reactions between R. parkeri, R. amblyommatis, and R. bellii, with endpoint titers between 128 and 512. A sample of 45 ticks collected from four culled wild boars was identified as Amblyomma sculptum, and all tested negative for rickettsial DNA presence. In conclusion, this study has provided a reliable sampling seroprevalence and indicated high exposure of wild boars to rickettsial agents, with a potential interaction with Rickettsia spp. from the spotted fever group within the Cerrado biome from midwestern Brazil.

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.

Amblyomma sculptum is a relevant tick species from a One Health perspective, playing an important role as a vector of Rickettsia rickettsii, the main agent of spotted fever rickettsiosis in Brazil. In this study, we evaluated the susceptibility of two A. sculptum populations from Goiás state (midwestern Brazil) to different acaricides. The first tick population (GYN strain) originated from an experimental farm, where the ticks are annually exposed to acaricides. The second (PNE strain) was collected in a national park (Emas National Park), where the ticks had not been exposed to acaricides. Immersion tests were conducted with 21-day-old laboratory-reared larvae and nymphs originating from adult ticks collected in the areas mentioned above. The chosen acaricides were two synthetic pyrethroids (cypermethrin and deltamethrin), one organophosphate (chlorfenvinphos), one formamidine (amitraz), and two combinations of pyrethroids and organophosphates (cypermethrin, chlorpyrifos and citronellal; cypermethrin, fenthion and chlorpyrifos). Mortality data were used to determine the lethal concentration (LC) values at which 50%, 90%, and 99% of the ticks died (LC50, LC90, and LC99, respectively), and resistance ratios (RR) were calculated based on the LC values. The RR revealed differences between the acaricide-exposed (GYN) and unexposed (PNE) tick strains. The PNE strain larvae and nymphs were susceptible to all the tested acaricides. The GYN strain larvae were tolerant to cypermethrin, whereas the nymphs were tolerant to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal (2 < RR ≤ 10). The GYN strain nymphs were resistant to amitraz (RR > 10). This is the first report of A. sculptum nymphs with resistance to amitraz and tolerance to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal.

Piroplasmids (order Piroplasmida) are a diverse group of tick-borne protozoa that may cause disease in animals and occasionally in humans. Novel Piroplasmida clades and species have been found in wild animals from Brazil based on the phylogenetic assessment of near-complete 18S rRNA, mitochondrial and heat-shock protein genes. For instance, a putative novel Babesia species has been detected in capybaras and Amblyomma ticks in three Brazilian states. The present work aimed to describe, using phylogenetic assessments based on distinct molecular markers, this novel Babesia species in capybaras and associated ticks (Amblyomma sculptum and Amblyomma dubitatum) sampled in Goiânia city, Goiás state, midwestern Brazil. While the phylogenetic analysis based on both near-complete 18S rRNA and hsp-70 genes positioned the sequences obtained from capybara blood samples into a new clade sister to the Babesia sensu stricto clade, the phylogenetic inference based on the COX-3 amino acid positioned the obtained sequences from capybara blood samples and A. sculptum ticks also into a clade sister to the Theileria sensu stricto clade, highlighting the inappropriateness of this marker inferring evolutionary relationships among piroplasmids. Pairwise distance analysis demonstrated that the divergence rates between the 18S rRNA sequences detected in capybaras and other Piroplasmida already described were very high and ranged from 9.4 to 12.9%. Genotype analysis based on the near-full 18S rRNA sequences of the Piroplasmida detected in capybaras and associated ticks demonstrated the occurrence of high genotype diversity at an intra-species level. In conclusion, phylogenetic analyses based on distinct molecular markers supported the description of Babesia goianiaensis nov. sp. in capybaras and associated Amblyomma ticks. Additionally, a novel phylogenetic clade, apart from the previously described ones, was described in the present study and contributed to untangling the complex evolutionary history of the Piroplasmida.

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

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: Ticks are significant parasites of dogs in the tropics, where tick-borne pathogens are highly prevalent, especially in areas where tick control measures are frequently neglected. This study investigated the seroprevalence and hematological abnormalities associated with Ehrlichia canis in dogs referred to a veterinary teaching hospital in Central-western Brazil. Out of 264 dogs tested for anti-Ehrlichia canis antibodies by an indirect immunofluorescence assay (IFA), 59.1% (156/264) were positive. Seropositivity was significantly associated to anemia and thrombocytopenia, alone or in combination, and to leukopenia. Conversely, there were no differences in terms of seroprevalence according to sex, breed and age. This study demonstrated that dogs referred to a veterinary teaching hospital in Central-western Brazil are highly exposed to E. canis and that seropositive dogs are more likely to present hematological abnormalities, particularly anemia, thrombocytopenia and leukopenia. To our knowledge, this is the first study on detection of anti-E. canis antibodies by means of IFA among dogs in the state of Goiás. These findings highlighted the need for increasing awareness among dog owners regarding tick control measures in Central-western Brazil, ultimately to reduce the risk of exposure to E. canis and other tick-borne pathogens.

O objetivo deste estudo foi descrever a distribuição dos nervos do membro pélvico do T. tetradactyla. Para tanto, foram utilizados quatro cadáveres de animais adultos cedidos pelo IBAMA-GO. Cada animal teve o membro pélvico de ambos os antímeros dissecados após fixação em solução de formaldeído a 10%. Os nervos genitofemoral e cutâneo femoral lateral distribuíram-se na pele medial e craniomedial e craniolateral da coxa, respectivamente. O nervo femoral emitiu ramos para os músculos psoas menor, iliopsoas, sartório e pectíneo; terminando por penetrar no músculo quadríceps femoral. Emitiu o nervo safeno, que em seu trajeto, inervou a pele da face medial da coxa e perna e região dorsomedial do pé. O nervo obturador emitiu ramos para músculos mediais da coxa. Os nervos glúteos cranial e caudal inervaram a musculatura glútea. O nervo isquiático inervou os músculos gêmeos, quadrado femoral, semi-membranoso, semitendinoso, bíceps femoral longo e curto e terminou dividindo-se nos nervos cutâneo lateral da sura, tibial, e fibular comum. O nervo cutâneo lateral da sura inerva a região cutânea craniolateral do joelho e perna, enquanto os nervos fibular comum e tibial inerva a musculatura da perna e terminam distribuindo-se no pé.