Rickettsial infection in ticks from a natural area of Atlantic Forest biome in southern Brazil.

From June 2013 to January 2014, blood sera samples and ticks were collected from domestic dogs and wild small mammals, and ticks from the vegetation in a preservation area of the Atlantic Forest biome (Turvo State Park), and the rural area surrounding the Park in Derrubadas municipality, state of Rio Grande do Sul, southern Brazil. Dogs were infested by Amblyomma ovale and Amblyomma aureolatum adult ticks, whereas small mammals were infested by immature stages of A. ovale, Amblyomma yucumense, Amblyomma brasiliense, Ixodes loricatus, and adults of I. loricatus. Ticks collected on vegetation were A. brasiliense, A. ovale, A. yucumense, Amblyomma incisum, and Haemaphysalis juxtakochi. Three Rickettsia species were molecularly detected in ticks: Rickettsia bellii in I. loricatus (also isolated through cell culture inoculation), Rickettsia amblyommatis in A. brasiliense, and Rickettsia rhipicephali in A. yucumense. The latter two are tick-rickettsia associations reported for the first time. Seroreactivity to Rickettsia antigens were detected in 33.5% (55/164) small mammals and 8.3% (3/36) canine sera. The present study reveals a richness of ticks and associated-rickettsiae in the largest Atlantic Forest Reserve of the state of Rio Grande do Sul, which is characterized by a rich fauna of wild mammals, typical of more preserved areas of this biome. Noteworthy, none of the detected Rickettsia species have been associated to human or animal diseases. This result contrasts to other areas of this biome in Brazil, which are endemic for tick-borne spotted fever caused by Rickettsia rickettsii or Rickettsia parkeri.

Seasonal dynamics of Amblyomma sculptum in two areas of the Cerrado biome midwestern Brazil, where human cases of rickettsiosis have been reported.

Amblyomma sculptum is a common human-biting tick in Brazil, where it plays an important role as a vector of Rickettsia rickettsii, the agent of the Brazilian spotted fever. Herein, we studied the seasonal dynamics of A. sculptum in an urban area of the Cerrado biome in midwestern Brazil, where human rickettsiosis is endemic. Ticks were collected in two sites located within the campus of Federal University of Goiás. The collections were done by dragging, flagging and visual search. In total, 117,685 ticks were collected, including 100,627 Amblyomma spp. larvae, 10,055 nymphs and 6977 adults of A. sculptum, and one nymph and 25 adults of Amblyomma dubitatum. The highest peak of larvae occurred in June 2018 and in July 2019, whereas nymphs peaked in July 2018 and September 2019. Adults reached their highest numbers in March 2018 and November 2019. These data suggest that A. sculptum develops one generation per year in this urban area of the Cerrado biome in midwestern Brazil. Interestingly, the peak of nymphs occurred during the same period of all confirmed cases of rickettsiosis in Goiás, suggesting a possible relationship between the seasonal dynamics of this tick stage and rickettsiosis transmission in this state.

One Health Approach on Ehrlichia canis: Serosurvey of Owners and Dogs, Molecular Detection in Ticks, and Associated Risk Factors in Tick-Infested Households of Southern Brazil.

Background: Ehrlichia canis has been the main hemopathogen affecting domestic dogs in Brazil. Even though tick-infested dogs may lead to household infestation and predispose human exposure and public health concern, no comprehensive study has surveyed humans, dogs, and environmental ticks altogether. Materials and Methods: Accordingly, the present study aimed to assess tick-infested households, identify tick species, perform serological (immunofluorescence assay) and molecular (PCR and q-PCR) detection of Ehrlichia in ticks, in the eighth biggest metropolitan area of Brazil. Results: Between 2007 and 2020, 233/5973 (3.9%) out of all complaints were from tick-infested households of 200 different addresses. Overall, 370/552 (67.0%) ticks were collected and identified as adult and 182/552 (33.0%) as immature forms of Rhipicephalus sanguineus s.l. complex; a single tick from one owner, a female tick of Amblyomma sculptum; and 395 ticks from dogs, 319/395 (80.8%) adult and 72/395 (18.2%) immature forms of Rhipicephalus spp., and 4/395 (1.01%) female Amblyomma aureolatum. Overall, 2/135 (1.5%) owners and 13/136 (9.6%) dogs were seropositive for E. canis. The DNA of Anaplasmataceae family was molecularly detected in 16/50 (32.0%) R. sanguineus s.l. As expected, the number of monthly tick infestation complaints were directly associated, and mean (p = 0.01), maximum (p = 0.011), and minimum (p = 0.008) temperature were statistically significant and had a low positive correlation (0.24, 0.23, and 0.24, respectively). In addition, complaints were highly associated to all socioeconomic variables (p < 0.001), with the exception of the presence of vacant lots. Conclusions: Despite low samplings and human negative results, areas with low-income with adequate temperature and urban agglomerations have been shown to be associated risks for tick infestations, predisposing tick-borne diseases. In conclusion, monitoring should always be conducted in such areas, including One Health approach with serosurvey of owners and dogs, along with identification and molecular screening of ticks.

Sensitivity of PCR in conjunctival swab samples for the diagnosis of canine visceral leishmaniasis: a systematic review and meta-analysis.

To compare the sensitivity of conjunctival swab (CS) and conventional samples (blood, spleen, liver, lymphoid and cutaneous tissue) in the diagnosis of canine visceral leishmaniasis (CVL) by polymerase chain reaction (PCR), a systematic review and meta-analysis was carried out using PubMed, Science Direct, Scopus, Web of Science, VHL/BVS (Virtual Health Library), CAPES, and Scielo databases. Articles published from 2002 to 2022 were considered and the review was updated in Jul 2023. From the total of 371 identified studies, 8 met all the eligibility criteria and were included in this review. Data from 658 CVL-positive dogs and 2541 PCR results were considered. Using a random effect model, data on the sensitivity of the test was compared between intervention (CS samples) and comparison (all the other samples) groups. Overall, the use of CS in the PCR diagnosis of CVL produced 12% higher sensitivity (p=0.013) in the test than all the other samples in combination. The animals' clinical condition did not influence (p>0.142) this overall result. However, when CS was individually compared to each of the conventional samples, the consistent result was observed (p=0.012) only in the CS versus bone marrow comparison. Given their rapid acquisition, minimal invasiveness, and lower cost relative to conventional samples, CS samples present a promising alternative for the molecular diagnosis of CVL.

First molecular detection of Borrelia theileri subclinical infection in a cow from Brazil.

Borrelia theileri is a relapsing fever group Borrelia that is transmitted to cattle by ticks of the genus Rhipicephalus. In this study, we describe the first molecular detection of B. theileri subclinical infection in a cow in Brazil. During the examination of stained blood smears of 10 cows from a farm with a recent history of fatal Trypanosoma vivax trypanosomiasis, spirochete-like structures were incidentally detected in one of the cows. The animal presented good body score, normal hematocrit and normal-colored ocular mucosa. Temperature, heart rate and respiratory rate were all normal. The animal was infested by ticks, which were morphologically identified as Rhipicephalus microplus. The diagnosis was confirmed by testing DNA extracted from a blood sample using a PCR targeting a ≈ 650 bp fragment of the flagellin B (flaB) gene of Borrelia spp. The partial flaB sequence obtained showed 99.83% similarity with B. theileri. Phylogenetically, the flaB partial sequence generated herein clustered with other B. theileri sequences, being separated from B. lonestari. This is the first molecular detection of B. theileri subclinical infection in a cow in Brazil. The possible implications of this finding are discussed.

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

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

Phylogenetic Inferences Based on Distinct Molecular Markers Confirm a Novel Babesia Species (Babesia goianiaensis nov. sp.) in Capybaras (Hydrochoerus hydrochaeris) and Associated Ticks.

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.

Rickettsial Infection in Ticks from a National Park in the Cerrado Biome, 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.

Susceptibility of Amblyomma sculptum, Vector of Rickettsia rickettsii, Ticks from a National Park and an Experimental Farm to Different Synthetic Acaricides.

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.

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.

Epidemiology of Rickettsia spp. in Atlantic rainforest areas of island and seashore mainland, southern Brazil.

Non-fatal cases of rickettsial infection with different clinical features than the classic BSF (Brazilian Spotted Fever) have been reported in seashore areas of Paraná state, southern Brazil. In addition, Amblyomma ovale tick infected by Rickettsia parkeri strain Atlantic rainforest has been also described in this area. Accordingly, the aim of this study was to investigate the occurrence of anti-Rickettsia spp. antibodies in human and dog populations, and Rickettsia spp. infection in ticks from oceanic islands and seashore mainland cities of southern Brazil. Serum samples were collected from 328 persons and their 282 dogs from three islands and two seashore mainland cities. A total of 211 ticks were collected from dogs, identified as A. ovale and R. sanguineus. In overall, 40 of 328 (12.2%) human samples were seropositive for Rickettsia spp., including 21 of 190 (11.1%) on islands and 19 of 138 (13.7%) on seashore mainland, and 62 of 282 (22.0%) dog samples, including 31 of 153 (20.3%) on islands and 31 of 129 (24.0%) in seashore mainland areas. In overall, nine of 82 (11.0%) ticks were positive to real-time PCR assay targeting a fragment of the rickettsial gltA gene, including two of 64 (3.1%) Rickettsia sanguineus and seven of 18 (38.9%) A. ovale, of which four were infected with the R. parkeri strain Atlantic rainforest. Despite no association between risk factors and Rickettsia spp. seropositivity was found in human beings, access to natural areas (p = .011) and tick infestation (p = .004) was significantly associated to dog seropositivity. The serological and molecular findings herein have confirmed previous tick and clinical case reports and enlarged the geographical occurrence of A. ovale infected by R. parkeri strain Atlantic rainforest in oceanic islands and seashore mainland cities of Paraná State, indicating a new likely transmission area of this new rickettsial infection in human beings and dogs of southern Brazil.

Detection of an Undescribed Babesia sp. in Capybaras and Amblyomma Ticks in Central-Western Brazil.

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.

Serosurvey on rickettsiae of the spotted fever group and Rickettsia bellii among dogs in the state of Goiás, Brazil.

The purpose of this study was to do a serological survey on three rickettsial species: Rickettsia rickettsii and Rickettsia parkeri, two species of the spotted fever group (SFG) that are considered to be great importance for public health; and Rickettsia bellii, a species of unknown pathogenicity that infects a variety of human-biting ticks. Serum samples from 273 dogs were tested using the indirect immunofluorescence assay (IFA). A total of 52 samples (19.04%) were seropositive for at least one of the three Rickettsia spp. antigens. Thirty-eight (73.07%), twelve (23.07%) and one (1.92%) of these dogs showed homologous reactions to R. bellii, R. rickettsii and R. parkeri, respectively. Our results showed that the seroprevalence of Rickettsia spp. was relatively low. However, the positive serological tests indicated that these dogs had become infected by these agents at some point in their lives. Lastly, our study adds to the previous knowledge on the epidemiology of rickettsiosis in the state of Goiás by doing the first record of detection of anti-R. rickettsii, R. parkeri and R. bellii antibodies by IFA among dogs, thus indicating that these agents may be circulating in the dog population analyzed.

Toxoplasma gondii in cattle in Brazil: a review.

Toxoplasma gondii is an apicomplexan protozoan that is frequently found in both humans and animals worldwide. The aim of this review was to list important aspects of Toxoplasma gondii infection in cattle in Brazil. The frequency of occurrence of T. gondii antibodies in Brazilian cattle ranges from 1 to 89.1%, depending on the region evaluated, based on data from 1978 to 2018. However, some characteristics of T. gondii infection in cattle remain uncertain, such as the role of meat intake in transmitting the parasite to humans. Most information regarding T. gondii infection among Brazilian cattle is limited to evaluations of the frequency of occurrence of antibodies. About 70% of the diagnoses of infection in these ruminants in Brazil are made via the indirect fluorescence antibody test (IFAT). Nevertheless, little is known about the population structure of this protozoan in cattle. It is necessary to expand the studies on toxoplasmosis in cattle, in order to better understand T. gondii infection in these animals and its implications for Brazilian public health.

Frequency of anti-Leptospira spp. antibodies in dogs and wild small mammals from rural properties and conservation units in southern Brazil.

Leptospirosis is a disease of worldwide distribution that affects man and several animal species. Domestic and wild animals can behave as reservoirs of the agent. The aim of this study was to evaluate the frequency of anti-Leptospira spp. antibodies in dogs and wild small mammals from rural properties and conservation units of three municipalities of Rio Grande do Sul State, Brazil. Sera were collected from 192 domestic dogs and 132 small mammals, namely rodents of the Cricetidae and Caviidae families and marsupials of the Didelphidae family. The study used Microscopic Agglutination Test (MAT) against 23 serovars of Leptospira spp. Overall, 9.90% (19/192) dogs, and 1.51% (2/132) small mammals were seropositive. Leptospira interrogans serovar Australis was the most frequent serovar in dogs and antibodies against L. interrogans serovar Pomona were detected in a dog and a small mammal belonging to the same municipality. We conclude that in the regions studied the frequency of seropositivity in dogs is low, and leptospires do not seem to be circulating in small mammals, yet, further research is necessary to assess the real role that these animals may have for leptospirosis in the studied areas. This is the first serological survey in small mammals, including wild rodents, in the area that encompasses conservation units of great importance to Rio Grande do Sul state.

Occurrence of Ehrlichia canis and Hepatozoon canis and probable exposure to Rickettsia amblyommatis in dogs and cats in Natal, RN.

Occurrence of infection or exposure to Ehrlichia canis, Hepatozoon canis and Rickettsia spp. was detected in feral cats living in two fragments from Atlantic rainforest, in Natal, RN, Brazil, and in dogs living around the parks. While serum samples were collected from 155 animals (53 cats living in the parks; 29 dogs living in human homes around the parks; and 73 dogs living at an animal control center - ACC), spleen samples were collected from 20 dogs that were euthanized at ACC. Serum samples were analyzed to Rickettsia spp. and E. canis antibodies using the indirect immunofluorescence assay. Seventeen of the 102 dogs (17%) had E. canis antibodies and 13% (20/155) of all dogs and cats (i.e. 3% (3/102) of the dogs and 32% (17/53) of the cats) were seropositive for Rickettsia spp. antigens. The animals were therefore been exposed to R. amblyommatis or by a very closely related genotype. Among the 20 dog spleen samples analyzed, eight were PCR positive for E. canis and two for H. canis (GenBank accession number MG772657 and MG772658, respectively). In none of the spleen samples were obtained amplicons for Babesia spp. through PCR. This study provided the first evidence that Rickettsia of the spotted fever group is circulating among dogs and cats in Natal.

Tick-borne infections in dogs and horses in the state of Espírito Santo, Southeast Brazil.

This work aims to identify and quantify the percentage of Rickettsia spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. positive pet dogs, and to identify ticks collected on these animals in the state of Espírito Santo, in the Southeast region of Brazil. The study included 378 dogs, 226 females and 152 males, of various breeds and ages (mean age of 4.1 years). All animals were examined for ticks, and whole blood was collected and processed by conventional PCR protocols for Babesia spp., Anaplasmataceae, Hepatozoon spp. and by real-time PCR for Ehrlichia canis. Of the 378 dogs examined, 157 (41.53%) had ticks at the time of the study, which were identified as R. sanguineus s.l. in 154 animals (98.10%), Amblyomma ovale in one animal (0.63%), and Amblyomma sculptum in 2 animals (1.27%). In the PCR for Babesia spp., 5 animals (1.32%) were positive, producing DNA sequences 100% identical to Babesia vogeli. For Anaplasmataceae, 34 animals (9%) were positive, 10 of which generated DNA sequences 100% similar to Ehrlichia canis. The other 24 samples generated fragments 100% identical to Anaplasma platys. In the PCR for Hepatozoon spp, 39 animals (10.31%) were positive, producing sequences 100% identical to Hepatozoon canis. Finally, in the real-time PCR specific for E. canis, 28 animals (7.40%) were positive. Coinfection with 2 or 3 agents was observed in 20 animals (5.29%). Of the 378 dogs sampled, 312 were analyzed by indirect immunofluorescence assay (IFA) for E. canis and five species of Rickettsia (R. rickettsii, R. parkeri, R. amblyommatis, R. rhipicephali and R. bellii). Among them, 71 dogs (22.75%) had a positive reaction for E. canis and 16 dogs (5.13%) had antibody titers higher than 64 to at least one Rickettsia species, 5 of them (1.60%) to R. rickettsii. Samples of blood were collected from 10 equines in the regions where dogs were found with positive serology for any one of the Rickettsia sp. tested. In the municipality of Vila Velha, two equines were positive for R. bellii with inverse titers of 64 and 128. These results indicate that tick-transmitted agents, such as E. canis, A. platys, B. vogeli, H. canis, and several spotted fever group Rickettsia are circulating in the canine population of the Brazilian state of Espirito Santo.

Litomosoides silvai (Nematoda: Onchocercidae) parasitizing Akodon montensis (Rodentia: Cricetidae) in the southern region of Brazil.

In the present study, Litomosoides silvai parasitizing Akodon montensis in the southern region of Brazil is reported for the first time. New morphological information is provided for some structures of this nematode species, such as a flattened cephalic extremity, presence of two dorsal cephalic papillae, female tail with a constriction at its tip, "s" shaped vagina, spicules characteristic of the carinii species group and microfilaria tail constricted at the tip. This nematode was found parasitizing the thoracic cavity with a prevalence of 10% (2/20), mean intensity of 4 (6/2), mean abundance of 0.4 (8/20) and range of infection of 2-6 specimens per host, in southern Brazil. This occurrence of L. silvai in A. montensis is a new geographical record for southern Brazil, in the Upper Paraná Atlantic Forest ecoregion of the northwestern region of Rio Grande do Sul, which is part of the Atlantic Forest biome.

Ehrlichia canis and Rickettsia spp. in dogs from urban areas in Paraiba state, northeastern Brazil.

The aims of our study was to identify Ehrlichia canis and antibodies against Rickettsia spp. belonging to the spotted fever group (SFG) in dogs sampled from Paraiba state, northeastern Brazil. Blood and serum samples collected by convenience from dogs in urban areas of five municipalities were analyzed by real-time PCR for the detection of E. canis DNA and by immunofluorescence assay test (IFAT) for the identification of antibodies against Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii and R. rhipicephali antigens. E. canis DNA was detected in 8.9% (64/719) of the blood samples, whereas 5.63% (43/763) of the serum samples were positive for at least one of the Rickettsia antigens tested by IFAT. This study showed for the first time the occurrence of E. canis and suggested the circulation of SFG Rickettsia in dogs in the study region of Paraiba state, northeastern Brazil.

Negative serosurvey of Toxoplasma gondii antibodies in Golden-headed Lion Tamarin (Leontopithecus chrysomelas) from Niterói/RJ, Brazil.

New World Nonhuman Primates are highly susceptible to clinical toxoplasmosis. Serum samples from 126 recently captured Leontopithecus chrysomelas, from an exotic and invasive population, were tested for Toxoplasma gondii antibodies by the modified agglutination test (MAT, cut-off 1:25); all were seronegative. The MAT is highly specific and is not species-specific. This is the first report of T. gondii survey in this tamarin in the wild. This result is consistent with prior reports that showed the high susceptibility of the species to infection by T. gondii usually with high mortality rates.