Phylogenetic inferences based on distinct molecular markers reveals a novel Babesia (Babesia pantanalensis nov. sp.) and a Hepatozoon americanum-related genotype in crab-eating foxes (Cerdocyon thous).

Piroplasmids and Hepatozoon spp. Are apicomplexan protozoa that may cause disease in several canid species. The present study aimed to expand the knowledge on the diversity of piroplasmids and Hepatozoon in crab-eating foxes (Cerdocyon thous; n = 12) sampled in the Pantanal of Mato Grosso do Sul State, central-western Brazil. PCR assays based on the 18S rRNA were used as screening. Three (25%) and 11 (91.7%) were positive for piroplasmids and Hepatozoon spp., respectively. Co-infection was found in three C. thous. Phylogenetic analyses based on the near-complete 18S rRNA, cox-1 and hsp70 genes evidenced the occurrence of a novel of Babesia spp. (namely Babesia pantanalensis nov. sp.) closely related to Rangelia vitalii and Babesia sp. 'Coco'. This finding was supported by the genetic divergence analysis which showed (i) high divergence, ranging from 4.17 to 5.62% for 18 S rRNA, 6.16% for hps70 and 4.91-9.25% for cox-1 and (ii) the genotype network (which displayed sequences separated from the previously described Piroplasmida species by median vectors and several mutational events). Also, phylogenetic analysis based on the 18S rRNA gene of Hepatozoon spp. positioned the sequences obtained herein in a clade phylogenetically related to Hepatozoon sp. 'Curupira 2', Hepatozoon sp. detected in domestic and wild canids from Uruguay and Hepatozoon americanum. The present study described Babesia pantanalensis nov sp. and Hepatozoon closely related to H. americanum in crab-eating foxes from Brazil. Moreover, the coinfection by piroplasmids and Hepatozoon sp. for the first time in crab-eating foxes strongly suggesting that this wild canid species potentially acts as a bio-accumulate of hemoprotozoan in wild environment.

Crithidia mellificae infection in different mammalian species in Brazil.

Crithidia mellificae, a monoxenous trypanosomatid considered restricted to insects, was recently reported to infect a bat. Herein, C. mellificae has been demonstrated to have a wider range of vertebrate hosts and distribution in Brazilian biomes than once thought. Parasites isolated from haemocultures were characterized using V7V8 SSU rDNA and glyceraldehyde 3-phosphate dehydrogenase genes. Coatis (Nasua nasua) in the Cerrado; marmosets (Callithrix sp.) and bats (Carollia perspicillata, Myotis lavali, M. izecksohni, Artibeus lituratus) in the Atlantic Forest; crab-eating foxes (Cerdocyon thous) and ocelot (Leopardus pardalis) in the Pantanal biomes were infected by trypanosomatids that displayed choanomastigote forms in haemoculture in Giemsa-stained slide smears. Molecular characterization and phylogenetic inference confirmed the infection of C. mellificae in these animals. Moreover, slight differences in C. mellificae sequences were observed. Crithidia mellificae growth curves were counted at 27°C, 36°C and 37°C, and the morphotypes were able to grow and survive for up to 16 days. Serological titers for C. mellificae were observed in nonhuman primates, demonstrating that this parasite is able to induce a humoral immune response in an infected mammal. These results showed that host specificity in trypanosomatids is complex and far from understood.

Uncovering Trypanosoma spp. diversity of wild mammals by the use of DNA from blood clots.

Trypanosoma spp. infection in wild mammals is detected mainly through parasitological tests that usually display low sensitivity. We propose the use of DNA extracted directly from blood clots (BC), which are neglected sources of DNA for diagnosis and identification of Trypanosoma spp. This approach followed by nested PCR targeting the 18S SSU rDNA demonstrated to be sensitive and suitable to evaluate the diversity of trypanosomes infecting sylvatic mammals, including subpatent and mixed infections. Infection was detected in 95/120 (79.2%) samples from bats, carnivores and marsupials that included negative serological and hemoculture testing mammals. Thirteen Trypanosoma spp. or Molecular Operational Taxonomic Units (MOTUs) were identified, including two new MOTUs. The high diversity of trypanosomes species and MOTUs infecting bats and marsupials showed that these hosts can be considered as bio-accumulators of Trypanosoma spp., with specimens of Didelphis spp. displaying the highest trypanosome diversity. The use of blood clots allowed direct access to non-culturable parasites, mixed infections, besides bypassing the selective pressure on the parasites inherent to cultivation procedures. Trypanosoma cruzi was the species found infecting the highest number of individuals, followed by T. lainsoni. Positive PCR for T. cruzi was observed in 16 seronegative individuals and 30 individuals with negative hemocultures. Also, T. lainsoni, previously found only in rodents, showed to be capable of infecting bats and marsupials. This finding makes it clear that some species of Trypanosoma are more generalist than previously thought. Molecular diagnosis using nested PCR from DNA extracted from BC allowed the increase of the knowledge about host-spectrum and distribution of Trypanosoma spp. and allowed the identification of new MOTUs.

Detection of Rangelia vitalii (Piroplasmida: Babesiidae) in asymptomatic free-ranging wild canids from the Pampa biome, Brazil.

Canine rangeliosis is a tick-borne disease caused by the protozoan Rangelia vitalii, which has only been reported in South America. With this knowledge, we hypothesized that neotropical foxes could act as asymptomatic natural carriers of R. vitalii. To test this, we captured 44 free-ranging foxes and investigated the presence of R. vitalii DNA, and whether the infected animals presented any clinical findings or hematological changes. Eight foxes (18%), seven Cerdocyon thous (7/27-25%), and one Lycalopex gymnocercus (1/17-5%) were positive for R. vitalii. All foxes were clinically healthy and showed no hematological abnormalities. Thus, we propose that neotropical canids, particularly C. thous, could be the natural carriers of R. vitalii.

Rickettsia spp. among wild mammals and their respective ectoparasites in Pantanal wetland, Brazil.

The genus Rickettsia comprises obligatory intracellular bacteria, well known to cause zoonotic diseases around the world. The present work aimed to investigate the occurrence of Rickettsia spp. in wild animals, domestic dogs and their respective ectoparasites in southern Pantanal region, central-western Brazil, by molecular and serological techniques. Between August 2013 and March 2015, serum, whole blood and/or spleen samples were collected from 31 coatis, 78 crab-eating foxes, seven ocelots, 42 dogs, 110 wild rodents, and 30 marsupials. Serum samples from canids, felids, rodents and marsupials were individually tested by indirect fluorescent antibody test (IFAT) in order to detect IgG antibodies to Rickettsia rickettsii, Rickettsia parkeri and Rickettsia amblyommatis. DNA samples from mammals and ectoparasites were submitted to a multiplex qPCR assay in order to detect and quantify spotted fever group (SFG) and typhus group (TG) rickettsiae and Orientia tsutsugamushi. Positive samples in qPCR assays were submitted to conventional PCR assays targeting gltA, ompA, ompB and htrA genes, followed by sequencing and phylogenetic analyses. The ticks collected (1582) from animals belonged to the species Amblyomma sculptum, Amblyomma parvum, Amblyomma ovale, Amblyomma tigrinum, Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus sensu lato and Amblyomma auricularium. Overall, 27 (64.2%) dogs, 59 (75.6%) crab-eating foxes and six (85.7%) ocelots were seroreactive (titer≥64) to at least one Rickettsia species. For 17 (40.4%) dogs, 33 (42.3%) crab-eating foxes, and two (33.3%) ocelots, homologous reactions to R. amblyommatis or a closely related organism were suggested. One hundred and sixteen (23.5%) tick samples and one (1.2%) crab-eating fox blood sample showed positivity in qPCR assays for SFG Rickettsia spp. Among SFG Rickettsia-positive ticks samples, 93 (80.2%) belonged to A. parvum, 14 (12%) belonged to A. sculptum species, three (2.5%) belonged to A. auricularim, and six (5.2%) were Amblyomma larval pools. Thirty samples out of 117 qPCR positive samples for SFG Rickettsia spp. also showed positivity in cPCR assays based on gltA, htrA and/or ompB genes. The Blast analyses showed 100% identity with 'Candidatus Rickettsia andeanae' in all 30 sequences obtained from gltA, htrA and/or ompB genes. The concatenated phylogenetic analysis based on gltA and 17-kDa htrA genes grouped the Rickettsia sequences obtained from tick samples in the same clade of 'Candidatus Rickettsia andeanae'. The present study revealed that wild and domestic animals in southern Pantanal region, Brazil, are exposed to SFG rickettsiae agents. Future studies regarding the pathogenicity of these agents are necessary in order to prevent human cases of rickettsiosis in Brazilian southern Pantanal.

High Triatoma brasiliensis Densities and Trypanosoma cruzi Prevalence in Domestic and Peridomestic Habitats in the State of Rio Grande do Norte, Brazil: The Source for Chagas Disease Outbreaks?

AbstractA total of 2,431 Triatoma brasiliensis were collected from 39 populations of Paraíba (PB) and Rio Grande do Norte (RN) states, Brazil. In PB, Trypanosoma cruzi infection was not detected in either peridomestic or domestic vector populations. In contrast, in RN, T. brasiliensis was detected with high parasite prevalence in these ecotopes (30.7-40.0%). Moreover, peridomicile insect population densities were more than double the average densities of all other settings evaluated (19.17 versus < 8.94 triatomine/man-hour). Genotyped parasites evidenced a mix of T. cruzi lineages circulating in both peridomestic and sylvatic populations. Although vector control efforts have dramatically decreased Chagas disease transmission to humans, recent outbreaks have been detected in four municipalities of RN state. Our results clearly evidence a worrisome proximity between infected vectors and humans in RN. Indeed, finding of infected T. brasiliensis inside homes is routinely recorded by local vector control surveillance staff around the outbreak area, challenging the current and conventional view that vector transmissions are controlled in northeastern Brazil. This scenario calls for strengthening vector control surveillance and interventions to prevent further Chagas transmission, especially in RN State.

Occurrence and molecular characterization of hemoplasmas in domestic dogs and wild mammals in a Brazilian wetland.

Hemotropic mycoplasmas are known to cause anemia in several mammalian species. The present work aimed to investigate the occurrence of Mycoplasma spp. in wild mammals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil. Between August 2013 and March 2015, 31 Nasua nasua, 78 Cerdocyon thous, seven Leopardus pardalis, 42 dogs, 110 wild rodents, and 30 marsupials were trapped and ectoparasites (ticks and fleas) found parasitizing the animals were collected. Mammals and ectoparasites DNA samples were submitted to conventional PCR assays for Mycoplasma spp. targeting 16S rRNA and RnaseP genes. Twenty-four N. nasua, three C. thous, two domestic dogs, one L. pardalis and one wild rodent were positive for 16S rRNA PCR protocols. Fourteen N. nasua samples were also positive in RnaseP PCR. No marsupial or arthropod showed positivity for Mycoplasma spp. The phylogenetic analyses based on 16S rRNA gene showed that all sequences obtained from dogs, two sequences obtained from C. thous and ten sequences obtained from N. nasua showed to be closely related to Mycoplasma haemocanis/Mycoplasma haemofelis species. Genotypes closely related to 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemomuris were detected in the L. pardalis and in the wild rodent, respectively. Probably a novel Mycoplasma genotype, closely related to a sequence obtained from a Brazilian capybara was detected in 14 N. nasua, based on a concatenated phylogenetic analysis of 16S rRNA and RnaseP genes. The present study revealed that wild animals in southern Pantanal region, Brazil, are exposed to different species of hemoplasmas.

Molecular detection of Hepatozoon spp. in domestic dogs and wild mammals in southern Pantanal, Brazil with implications in the transmission route.

Hepatozoon parasites comprise intracellular apicomplexan parasites transmitted to vertebrate animals by ingestion of arthropods definitive hosts. The present work aimed to investigate the occurrence of Hepatozoon spp. in wild animals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil, by molecular techniques. Between August 2013 and March 2015, 31 coatis (Nasua nasua), 78 crab-eating foxes (Cerdocyon thous), seven ocelots (Leopardus pardalis), 42 dogs (Canis lupus familiaris), 110 wild rodents (77 Thichomys fosteri, 25 Oecomys mamorae, and 8 Clyomys laticeps), 30 marsupials (14 Thylamys macrurus, 11 Gracilinanus agilis, 4 Monodelphis domestica and 1 Didelphis albiventris), and 1582 ticks and 80 fleas collected from the sampled animals were investigated. DNA samples were submitted to PCR assays for Hepatozoon spp. targeting 18S rRNA gene. Purified amplicons were directly sequenced and submitted to phylogenetic analysis. A high prevalence of Hepatozoon among carnivores (C. thous [91.02%], dogs [45.23%], N. nasua [41.9%] and L. pardalis [71.4%]) was found. However, ticks and fleas were negative to Hepatozoon PCR assays. By phylogenetic analysis based on 18S rRNA sequences, Hepatozoon sequences amplified from crab-eating foxes, dogs, coatis and ocelots clustered with sequences of H. canis, H. americanum and H. felis. The closely related positioning of Hepatozoon sequences amplified from wild rodents and T. macrurus marsupial to Hepatozoon from reptiles and amphibians suggest a possible transmission of those Hepatozoon species between hosts by ectoparasites or by predation. Hepatozoon haplotypes found circulating in wild rodents seem to present a higher degree of polymorphism when compared to those found in other groups of animals. Although rodents seem not to participate as source of Hepatozoon infection to wild carnivores and domestic dogs, they may play an important role in the transmission of Hepatozoon to reptiles and amphibians in Pantanal biome.

Bats in urban areas of Brazil: roosts, food resources and parasites in disturbed environments.

Urbanization is a widespread intense land use that generally results in biodiversity decline. Among the taxa capable to adapt to urban landscapes, bats are particularly ubiquitous. Brazil has one of the world's largest diversity of bat species and one of the highest urbanization rates of the world. Yet, few studies have synthesized the biology of bats in urban environments, especially in Brazil. To fill this gap, we systematically reviewed the published scientific literature on the bat fauna found in urban areas of Brazil. The knowledge of urban bats is still incipient and heterogeneously spatially distributed, mostly concentrated in the southeastern region of the country. The assembled list of 84 urban species, of which nineteen are new species records for urban areas (including one new family), represents 47% of the bat richness registered in the country. Thirty-one bat species (37%) were captured exclusively inside forest fragments. Moreover, we provide information on the resources used within the urban matrix by summarizing the roosting sites for 38 bat species, as well as 31 plants consumed by at least twelve bat species. Regarding parasitological aspects, we listed eleven zoonotic parasites hosted by 27 bat species and discussed their potential to become a public health threat. Likewise, we considered the different features linked to urbanization, including impacts on immunity, body condition and susceptibility to acquiring parasites, as possible bat conservation issues. Finally, we defined an agenda for bat studies in urban areas of Brazil.

Complexity and multi-factoriality of Trypanosoma cruzi sylvatic cycle in coatis, Nasua nasua (Procyonidae), and triatomine bugs in the Brazilian Pantanal.

BACKGROUND: Trypanosoma cruzi is dispersed in nature through many transmission mechanisms among a high diversity of vectors and mammalian species, representing particular behaviors and habitats. Thus, each locality has a unique set of conditions underlying the maintenance of this parasite in the wild. The aim of the present study was to evaluate the life-cycle of T. cruzi in free-ranging coatis from the central region of the Brazilian Pantanal using a multi-factorial approach. METHODS: Three methodological blocks were used in the present study: (i) We evaluated T. cruzi infection using serological (ELISA) and parasitological (hemoculture) tests in free-ranging coatis captured from October 2010 to March 2012. In addition, we characterized T. cruzi isolates as DTUs (Discrete Typing Units); (ii) We evaluated Trypanosoma infection in species of Triatoma and Rhodnius inhabiting coati arboreal nests from May to September 2012 using parasitological and molecular assays; and (iii) We analyzed a set of longitudinal data (from 2005 to 2012) concerning the effects of T. cruzi infection on this coati population. Herein, we investigated whether seasonality, host sex, and host age influence T. cruzi prevalence and patterns of infection. RESULTS: The 2010-2012 period presented high seroprevalence on coatis (72.0 % ELISA) and a high percentage of individuals with infectivity competence (20.5 % positive hemoculture). All isolates presented TcI band patterns, occurring in single (n = 3) and mixed infections (1 TcI/T. rangeli; 4 with undefined characterization). Both male and female individuals presented the same transmission potential, expressed as positive hemoculture, which was only detected in the summer. However, overall, the data (2005-2012) highlighted the importance of females for T. cruzi maintenance in the winter. Moreover, twenty-three (67.6 %) bugs from five coati nests (71.4 %) were infected with flagellated forms. Seventeen samples were characterized as T. cruzi (TcI and TcIII genotypes). CONCLUSION: In the Pantanal region, T. cruzi is transmitted in a complex, multifactorial, dynamic and non-linear transmission web. The coati nests might be inserted in this web, acting as important transmission foci at the arboreal stratum to different mammal species with arboreal or scansorial behavior.

Infestation of arboreal nests of coatis by triatomine species, vectors of Trypanosoma cruzi, in a large Neotropical wetland.

The coati (Nasua nasua, Carnivora) is a medium-sized mammal common in the Pantanal of Brazil. Unlike most mammals, coatis construct arboreal nests used for resting and reproduction. In this region, the coati is an important host of Trypanosoma cruzi, the causative agent of Chagas disease. There are two possible routes through coatis can be infected by T. cruzi: the oral route or the vectorial route. However, the relative importance of each of these routes in the infection of coatis and its role in the sylvatic cycle of the parasite are unknown. Our objectives were to investigate: (i) whether coati nests were infested by triatomine bugs, (ii) what species were frequent in the nests, (iii) whether the triatomines in nests were infected by T. cruzi, and (iv) what were the food resources of these triatomines. Eight of the 24 nests sampled were infested with triatomines, a total of 37 specimens of at least two species (Rhodnius stali and Triatoma sordida). In one nest, R. stali and T. sordida co-occurred and both fed on multiple resources, including coatis. This is the first report of triatomines occurring in arboreal nests of coatis. The co-occurrence of two different genera of triatomine vectors and coatis within the limited space of the coati nests provide multiple opportunities for the exchange of the protozoan parasite through both the vectorial and oral transmission routes.

Carrapatos (Acari: Ixodidae) em mamíferos silvestres do Parque Nacional da Serra da Canastra e arredores, Minas Gerais, Brasil / Ticks (Acari: Ixodidae) on wild mammals in the Serra da Canastra National Park and surrounding areas, Minas Gerais, Brazil

O presente estudo relata ixodídeos em mamíferos silvestres no Parque Nacional da Serra da Canastra e arredores, no estado de Minas Gerais, Brasil. De julho de 2005 a junho de 2014, foram coletadas 58 larvas, 133 ninfas e 43 carrapatos adultos em 40 eventos de captura de mamíferos de vida livre do Parque e arredores. Nos hospedeiros da ordem Carnivora, foram identificados Amblyomma ovale Koch, 1844, Amblyomma sculptum Berlese, 1888, Amblyomma tigrinum Koch, 1844, Dermacentor nitens Neumann, 1897 e Rhipicephalus microplus (Canestrini, 1888). Em hospedeiros da ordem Xenarthra, foram identificados espécimes de Amblyomma pseudoconcolor Aragão, 1908 e A. sculptum. Este estudo relata os primeiros registros dos ixodídeos D. nitens e R. microplus parasitando Lycalopex vetulus, e A. ovale em Leopardus tigrinus no país. Os achados do presente estudo indicam a proximidade entre os animais domésticos e silvestres causada pela expansão agropecuária no território brasileiro.(AU)

This research­­­­­ reports ticks on wild mammals from the Serra da Canastra National Park and surrounding areas, in Minas Gerais state, Brazil. From July 2005 to June 2014, 58 larvae, 133 nymphs and 43 adult ticks were collected in 40 capture events of free-living mammals at the Park and surrounding areas. The tick species Amblyomma ovale Koch, 1844, Amblyomma sculptum Berlese, 1888, Amblyomma tigrinum Koch, 1844, Dermacentor nitens Neumann, 1897, and Rhipicephalus microplus (Canestrini, 1888) were identified on hosts of the order Carnivora. Specimens of Amblyomma pseudoconcolor Aragão, 1908 and A. sculptum were identified on the hosts of the order Xenarthra. We provide the first records of D. nitens and R. microplus ticks parasitizing Lycalopex vetulus, and A. ovale on Leopardus tigrinus in Brazil. The findings of this research indicate the sharing of environment by domestic and wild animals caused by agricultural expansion in the Brazilian landscape.(AU)

Carrapatos (Acari: Ixodidae) em mamíferos silvestres do Parque Nacional da Serra da Canastra e arredores, Minas Gerais, Brasil / Ticks (Acari: Ixodidae) on wild mammals in the Serra da Canastra National Park and surrounding areas, Minas Gerais, Brazil

O presente estudo relata ixodídeos em mamíferos silvestres no Parque Nacional da Serra da Canastra e arredores, no estado de Minas Gerais, Brasil. De julho de 2005 a junho de 2014, foram coletadas 58 larvas, 133 ninfas e 43 carrapatos adultos em 40 eventos de captura de mamíferos de vida livre do Parque e arredores. Nos hospedeiros da ordem Carnivora, foram identificados Amblyomma ovale Koch, 1844, Amblyomma sculptum Berlese, 1888, Amblyomma tigrinum Koch, 1844, Dermacentor nitens Neumann, 1897 e Rhipicephalus microplus (Canestrini, 1888). Em hospedeiros da ordem Xenarthra, foram identificados espécimes de Amblyomma pseudoconcolor Aragão, 1908 e A. sculptum. Este estudo relata os primeiros registros dos ixodídeos D. nitens e R. microplus parasitando Lycalopex vetulus, e A. ovale em Leopardus tigrinus no país. Os achados do presente estudo indicam a proximidade entre os animais domésticos e silvestres causada pela expansão agropecuária no território brasileiro.

This research­­­­­ reports ticks on wild mammals from the Serra da Canastra National Park and surrounding areas, in Minas Gerais state, Brazil. From July 2005 to June 2014, 58 larvae, 133 nymphs and 43 adult ticks were collected in 40 capture events of free-living mammals at the Park and surrounding areas. The tick species Amblyomma ovale Koch, 1844, Amblyomma sculptum Berlese, 1888, Amblyomma tigrinum Koch, 1844, Dermacentor nitens Neumann, 1897, and Rhipicephalus microplus (Canestrini, 1888) were identified on hosts of the order Carnivora. Specimens of Amblyomma pseudoconcolor Aragão, 1908 and A. sculptum were identified on the hosts of the order Xenarthra. We provide the first records of D. nitens and R. microplus ticks parasitizing Lycalopex vetulus, and A. ovale on Leopardus tigrinus in Brazil. The findings of this research indicate the sharing of environment by domestic and wild animals caused by agricultural expansion in the Brazilian landscape.

Serology for brucellosis in free-ranging crab-eating foxes (Cerdocyon thous) and brown-nosed coatis (Nasua nasua) from Brazilian Pantanal / Sorologia para brucelose em lobinhos (Cerdocyon thous) e coatis (Nasua nasua) de vida livre do Pantanal brasileiro

A serological survey in free-ranging crab-eating foxes (Canidae: Cerdocyon thous) and brown-nosed coatis (Procyonidae: Nasua nasua) was performed in the Nhecolândia sub-region of the Brazilian Pantanal to evaluate the presence of anti-smooth Brucella antibodies on those wild populations. The detection of anti-smooth Brucella antibodies was performed by the Rose Bengal Test (RBT) as screening test and the Fluorescence Polarization Assay (FPA) as a confirmatory test. The frequency of smooth Brucella seropositive animals were 13.2% (5/38, 95% CI: 4.4% - 28.1%) for crab-eating foxes and 8.8% (3/34, 95% CI: 1.9% -23.7%) for brown-nosed coatis. No association was found between seropositivity for brucellosis and gender or age. The results of this study suggest exposure to or infection of crab-eating fox and brown-nosed coati from the Brazilian Pantanal by Brucella spp.(AU)

Um inquérito sorológico em quatis (Procyonidae: Nasua nasua) e lobinhos (Canidae: Cerdocyon thous) de vida livre foi conduzido na sub-região da Nhecolândia do Pantanal Sulmatogrossense para avaliar a presença de anticorpos anti-Brucella (fenótipo liso). A pesquisa de anticorpos anti-Brucella foi realizada pelo teste do antígeno acidificado tamponado, utilizado como teste de triagem e pelo teste de polarização fluorescente, utilizado como teste confirmatório. A frequência de animais soropositivos para anticorpos anti-Brucella foi de 13,2% (5/38, IC 95%: 4,4% - 28,1%) para lobinhos e de 8,8% (3/34, IC 95%: 1,9% - 23,7%) para coatis. Não foi encontrada associação entre a soropositividade para brucelose e gênero ou idade. Os resultados deste estudo sugerem exposição a ou infecção de lobinhos e quatis do Pantanal Brasileiro por Brucella spp.(AU)

Serology for brucellosis in free-ranging crab-eating foxes (Cerdocyon thous) and brown-nosed coatis (Nasua nasua) from Brazilian Pantanal / Sorologia para brucelose em lobinhos (Cerdocyon thous) e coatis (Nasua nasua) de vida livre do Pantanal brasileiro

A serological survey in free-ranging crab-eating foxes (Canidae: Cerdocyon thous) and brown-nosed coatis (Procyonidae: Nasua nasua) was performed in the Nhecolândia sub-region of the Brazilian Pantanal to evaluate the presence of anti-smooth Brucella antibodies on those wild populations. The detection of anti-smooth Brucella antibodies was performed by the Rose Bengal Test (RBT) as screening test and the Fluorescence Polarization Assay (FPA) as a confirmatory test. The frequency of smooth Brucella seropositive animals were 13.2% (5/38, 95% CI: 4.4% - 28.1%) for crab-eating foxes and 8.8% (3/34, 95% CI: 1.9% -23.7%) for brown-nosed coatis. No association was found between seropositivity for brucellosis and gender or age. The results of this study suggest exposure to or infection of crab-eating fox and brown-nosed coati from the Brazilian Pantanal by Brucella spp.

Um inquérito sorológico em quatis (Procyonidae: Nasua nasua) e lobinhos (Canidae: Cerdocyon thous) de vida livre foi conduzido na sub-região da Nhecolândia do Pantanal Sulmatogrossense para avaliar a presença de anticorpos anti-Brucella (fenótipo liso). A pesquisa de anticorpos anti-Brucella foi realizada pelo teste do antígeno acidificado tamponado, utilizado como teste de triagem e pelo teste de polarização fluorescente, utilizado como teste confirmatório. A frequência de animais soropositivos para anticorpos anti-Brucella foi de 13,2% (5/38, IC 95%: 4,4% - 28,1%) para lobinhos e de 8,8% (3/34, IC 95%: 1,9% - 23,7%) para coatis. Não foi encontrada associação entre a soropositividade para brucelose e gênero ou idade. Os resultados deste estudo sugerem exposição a ou infecção de lobinhos e quatis do Pantanal Brasileiro por Brucella spp.

Identificação de mamíferos silvestres do Pantanal sul-mato-grossense portadores de Leptospira spp / Identification of wild mammals from Pantanal sul-matogrossense carriers of Leptospira spp

Foi realizado um levantamento da infecção por Leptospira spp. em mamíferos silvestres do Pantanal sul-mato-grossense com o emprego da reação de soroaglutinação microscópica (SAM) e da reação em cadeia da polimerase (PCR). Os sorovares de maior frequência nos animais investigados foram Hardjobovis (28%), Icterohemorhagiae (12%), M-110/2006 (isolado de Cerdocyon thous; 16%), Canicola (L014 isolada de Bos taurus, 4%), Whitcombi (4%), Pomona (20%), Autumnalis (12%) e Copenhageni (M9/99 isolada de Rattus norvegicus, 4%). Das 79 amostras examinadas pela PCR, 21 (26,58%) foram positivas, com a amplificação de um fragmento de aproximadamente 331pb. Dois fragmentos amplificados obtidos de amostras de C. thous foram clonados, sequenciados e identificados como L. interrogans por análise filogenética.

A survey of Leptospira spp. in wild mammals from the southern Pantanal of Mato Grosso do Sul was performed by microscopic agglutination test (MAT) and polymerase chain reaction (PCR). The serovars most frequently found were Hardjobovis (28%), Icterohemorhagiae (12%), M110/2006 strain (isolated from Cerdocyon thous, 16%), Canicola (L014 isolated from Bos Taurus, 4%), Whitcombi (4%), Pomona (20%), Autumnalis (12%) and Copenhageni (M9/99 isolated from Rattus norvegicus, 4%). From the 79 samples tested by PCR, 21 (26.58%) were positive, resulting in the amplification fragment of approximately 331pb. Two amplified fragments from C. thous were cloned, sequenced and identified as L. interrogans by phylogenetic analysis.

Identificação de mamíferos silvestres do Pantanal sul-mato-grossense portadores de Leptospira spp / Identification of wild mammals from Pantanal sul-matogrossense carriers of Leptospira spp

Foi realizado um levantamento da infecção por Leptospira spp. em mamíferos silvestres do Pantanal sul-mato-grossense com o emprego da reação de soroaglutinação microscópica (SAM) e da reação em cadeia da polimerase (PCR). Os sorovares de maior frequência nos animais investigados foram Hardjobovis (28%), Icterohemorhagiae (12%), M-110/2006 (isolado de Cerdocyon thous; 16%), Canicola (L014 isolada de Bos taurus, 4%), Whitcombi (4%), Pomona (20%), Autumnalis (12%) e Copenhageni (M9/99 isolada de Rattus norvegicus, 4%). Das 79 amostras examinadas pela PCR, 21 (26,58%) foram positivas, com a amplificação de um fragmento de aproximadamente 331pb. Dois fragmentos amplificados obtidos de amostras de C. thous foram clonados, sequenciados e identificados como L. interrogans por análise filogenética.(AU)

A survey of Leptospira spp. in wild mammals from the southern Pantanal of Mato Grosso do Sul was performed by microscopic agglutination test (MAT) and polymerase chain reaction (PCR). The serovars most frequently found were Hardjobovis (28%), Icterohemorhagiae (12%), M110/2006 strain (isolated from Cerdocyon thous, 16%), Canicola (L014 isolated from Bos Taurus, 4%), Whitcombi (4%), Pomona (20%), Autumnalis (12%) and Copenhageni (M9/99 isolated from Rattus norvegicus, 4%). From the 79 samples tested by PCR, 21 (26.58%) were positive, resulting in the amplification fragment of approximately 331pb. Two amplified fragments from C. thous were cloned, sequenced and identified as L. interrogans by phylogenetic analysis.(AU)

Trypanosoma cruzi infection in neotropical wild carnivores (Mammalia: Carnivora): at the top of the T. cruzi transmission chain.

Little is known on the role played by Neotropical wild carnivores in the Trypanosoma cruzi transmission cycles. We investigated T. cruzi infection in wild carnivores from three sites in Brazil through parasitological and serological tests. The seven carnivore species examined were infected by T. cruzi, but high parasitemias detectable by hemoculture were found only in two Procyonidae species. Genotyping by Mini-exon gene, PCR-RFLP (1f8/Akw21I) and kDNA genomic targets revealed that the raccoon (Procyon cancrivorus) harbored TcI and the coatis (Nasua nasua) harbored TcI, TcII, TcIII-IV and Trypanosoma rangeli, in single and mixed infections, besides four T. cruzi isolates that displayed odd band patterns in the Mini-exon assay. These findings corroborate the coati can be a bioaccumulator of T. cruzi Discrete Typing Units (DTU) and may act as a transmission hub, a connection point joining sylvatic transmission cycles within terrestrial and arboreal mammals and vectors. Also, the odd band patterns observed in coatis' isolates reinforce that T. cruzi diversity might be much higher than currently acknowledged. Additionally, we assembled our data with T. cruzi infection on Neotropical carnivores' literature records to provide a comprehensive analysis of the infection patterns among distinct carnivore species, especially considering their ecological traits and phylogeny. Altogether, fifteen Neotropical carnivore species were found naturally infected by T. cruzi. Species diet was associated with T. cruzi infection rates, supporting the hypothesis that predator-prey links are important mechanisms for T. cruzi maintenance and dispersion in the wild. Distinct T. cruzi infection patterns across carnivore species and study sites were notable. Musteloidea species consistently exhibit high parasitemias in different studies which indicate their high infectivity potential. Mesocarnivores that feed on both invertebrates and mammals, including the coati, a host that can be bioaccumulator of T. cruzi DTU's, seem to take place at the top of the T. cruzi transmission chain.

Trypanosoma cruzi TcI and TcII transmission among wild carnivores, small mammals and dogs in a conservation unit and surrounding areas, Brazil.

Aiming to better understand the ecological aspects of Trypanosoma cruzi transmission cycles, wild carnivores, small mammals and dogs were examined for T. cruzi infection in the Serra da Canastra National Park region, Brazil. Isolates were genotyped using mini-exon gene and PCR-RFLP (1f8 and H3) genomic targets. Trypanosoma cruzi transmission was well established in the area and occurred in both wild and peridomestic environments. Dog seroprevalence was 29·4% (63/214) and TcI and TcII genotypes, besides mixed infections were observed. Only TcI was detected in wild mammals. Marsupials displayed lower relative abundance, but a high prevalence of positive haemocultures (4/22), whereas rodents displayed positive haemocultures (9/113) mainly in the abundant Akodon montensis and Cerradomys subflavus species. The felid Leopardus pardalis was the only carnivore to display positive haemoculture and was captured in the same region where the small mammal prevalence of T. cruzi infection was high. Two canid species, Chrysocyon brachyurus and Cerdocyon thous, were serologically positive for T. cruzi infection (4/8 and 8/39, respectively), probably related to their capacity to exploit different ecological niches. Herein, dog infection not only signals T. cruzi transmission but also the genotypes present. Distinct transmission strategies of the T. cruzi genotypes are discussed.