RESUMO
Coatis (Nasua nasua) are wild carnivorous well adapted to anthropized environments especially important because they act as reservoirs hosts for many arthropod-borne zoonotic pathogens. Information about filarioids from coatis and associated Wolbachia spp. in Brazil is scant. To investigate the diversity of filarial nematodes, blood samples (n = 100 animals) were obtained from two urban areas in midwestern Brazil and analyzed using blood smears and buffy coats and cPCR assays based on the cox1, 12S rRNA, 18S rRNA, hsp70 and myoHC genes for nematodes and 16S rRNA for Wolbachia. When analyzing coati blood smears and buffy coats, 30% and 80% of the samples presented at least one microfilaria, respectively. Twenty-five cox1 sequences were obtained showing 89% nucleotide identity with Mansonella ozzardi. Phylogenetic analyses clustered cox1 sequences herein obtained within the Mansonella spp. clade. Sequences of both myoHC and two hsp70 genes showed 99.8% nucleotide identity with Mansonella sp. and clustered into a clade within Mansonella sp., previously detected in coatis from Brazil. Two blood samples were positive for Wolbachia, with a 99% nucleotide identity with Wolbachia previously found in Mansonella perstans, Mansonella ozzardi and Mansonella atelensis and in ectoparasites of the genus Pseudolynchia, Melophagus and Cimex. The study showed a high prevalence of Mansonella sp. in the coati population examined, suggesting that this animal species play a role as reservoirs of a novel, yet to be described, species within the Onchocercidae family.
RESUMO
Procyonids are reservoirs of many zoonotic infectious diseases, including tick-borne pathogens. The role of coatis (Nasua nasua) in the epidemiology of piroplasmids and Rickettsia has not been fully addressed in Brazil. To molecularly study these agents in coatis and associated ticks, animals were sampled in two urban areas in Midwestern Brazil. Blood (n = 163) and tick (n = 248) DNA samples were screened by PCR assays targeting the 18S rRNA and gltA genes of piroplasmids and Rickettsia spp., respectively. Positive samples were further molecularly tested targeting cox-1, cox-3, ß-tubulin, cytB, and hsp70 (piroplasmid) and ompA, ompB, and htrA 17-kDa (Rickettsia spp.) genes, sequenced and phylogenetically analyzed. All coatis' blood samples were negative for piroplasmids, whereas five pools of ticks (2%) were positive for two different sequences of Babesia spp.. The first from Amblyomma sculptum nymphs was close (i.e., ≥ 99% nucleotide identity) to a Babesia sp. previously found in capybaras (Hydrochoerus hydrochaeris); the second from Amblyomma dubitatum nymphs and Amblyomma spp. larvae was identical (100% nucleotide identity) to a Babesia sp. detected in opossums (Didelphis albiventris) and associated ticks. Four samples (0.8%) were positive by PCR to two different Rickettsia spp. sequences, being the first from Amblyomma sp. larva identical to Rickettsia belli and the second from A. dubitatum nymph identical to Rickettsia species from Spotted Fever Group (SFG). The detection of piroplasmids and SFG Rickettsia sp. highlights the importance of Amblyomma spp. in the maintenance of tick-borne agents in urban parks where humans and wild and domestic animals are living in sympatry.
Assuntos
Babesia , Ixodidae , Procyonidae , Rickettsia , Carrapatos , Humanos , Animais , Rickettsia/genética , Babesia/genética , Brasil/epidemiologia , Roedores , Gambás , Amblyomma , Ixodidae/microbiologiaRESUMO
The present study aimed to detect molecularly the presence of co-infections by vector-borne agents (VBA) in ring-tailed coatis' (Nasua nasua) blood samples from Iguaçu National Park (INP), southern Brazil, and assess the phylogenetic positioning of the detected agents. DNA blood samples were submitted to molecular screening and characterization for Anaplasmataceae agents, Piroplasmids, Hepatozoon sp., hemotropic mycoplasmas, and Bartonella spp. In total, 42 (85.7%) coatis were positive for hemotropic Mycoplasma sp., 12 (24.5%) for Bartonella machadoae, 7 (14.3%) for Anaplasma sp. closely related to 'Candidatus Anaplasma brasiliensis', and 3 (6%) for Hepatozoon procyonis. The most prevalent co-infections observed was from bacterial VBA: while 18.3% were co-infected by hemotropic Mycoplasma sp. and Bartonella sp., 12.2% were co-infected by Anaplasma sp. and hemotropic Mycoplasma sp. Only two animals (4%) presented co-infections by three VBA (Bartonella sp., Anaplasma sp. and hemotropic Mycoplasma sp.). The coati is a wild carnivore found in INP, mainly in areas visited by tourists. These animals are frequently seen searching for food in garbage dumps or in tourists' belongings. The present study expands the host specificity range of B. machadoae, which has been isolated only from rodents until the present moment. Since the zoonotic potential and transmission routes of the detected VBA are not yet known, surveillance in this area is much needed.
Assuntos
Bartonella , Coinfecção , Mycoplasma , Procyonidae , Animais , Procyonidae/microbiologia , Brasil/epidemiologia , Filogenia , Coinfecção/epidemiologia , Parques Recreativos , Bartonella/genética , Anaplasma/genéticaRESUMO
A tripanossomíase bovina é causada pelo protozoário Trypanosoma vivax. A transmissão biológica ocorre apenas no continente africano pela mosca Tsé-tsé, de forma mecânica por dípteros hematófagos em todos os continentes, ou pelo compartilhamento de agulhas e por práticas associadas. O estudo teve como objetivo relatar o primeiro diagnóstico parasitológico, sorológico e molecular de T. vivax em bovinos leiteiros provenientes de cinco propriedades do município de Unai, Minas Gerais, Brasil. Cento e quinze animais selecionados por conveniência apresentavam sinais clínicos ou pertenciam a lotes de animais suspeitos. Foram detectados positivos pelos testes parasitológico (técnica de Woo), sorológico (ELISA) e molecular (LAMP). A maior prevalência global para T. vivax foi de 11,11% na propriedade A. O único sinal clínico dos animais positivos estudados foi baixa taxa de concepção. O primeiro diagnóstico de tripanossomíase no noroeste mineiro é extremamente importante, haja vista o tamanho do rebanho leiteiro da região e as possíveis perdas econômicas provocadas pela enfermidade. Ademais, faz-se necessário maior controle sanitário na região, uma vez que a transmissão no Brasil é intimamente ligada às práticas de compartilhamento de agulhas no manejo dos animais e ao parasitismo de moscas hematófagas.
Assuntos
Animais , Bovinos , Tripanossomíase Bovina/diagnóstico , Trypanosoma vivax/isolamento & purificação , Brasil , Ensaio de Imunoadsorção Enzimática/veterinária , Transcrição ReversaRESUMO
This study aimed to morphologically and molecularly detect Hepatozoon procyonis in ring-tailed coatis' (Nasua nasua) blood and associated ticks from central-western Brazil, Campo Grande, Mato Grosso do Sul state and also evaluate the impact of the protozoa in blood parameters and coati´s health. Samplings were performed in a conservation area Parque Estadual do Prosa (PEP) and in a Brazilian Air Force Private Area namely Vila da Base Aérea (VBA), between March 2018 and April 2019. We collected 165 blood samples, 61 from recaptured coatis. Peripheral blood smears were stained with Romanovsky-type stain for H. procyonis parasitemia assessment. DNA extracted from blood samples and ticks (Amblyomma spp.) were submitted to a nested PCR (nPCR) assay based on the 18S rRNA gene for Hepatozoon spp. Out of 104 individuals sampled, 80 (77%) were positive for H. procyonis in at least one capture. Overall, 67/165 (40.6%) blood smears showed H. procyonis gametocytes (PEP: 41/63 - 65%; VBA: 26/102 - 25.5%). Parasitemia based on 500 assessed leucocytes ranged from 1 (0.2%) to 50 (10%) and 1 (0.2%) to 25 (5%), from animals sampled in PEP and VBA, respectively. Fluctuation on the parasitemia was observed during recaptures. nPCR results showed higher positivity when compared to blood smears, i.e. 112/165 (68%) positive blood samples [PEP: 41/63 (65%), VBA: 26/102 (25.5%)]. In total, 63/248 (25.4%) tick DNA samples were positive at nPCR for Hepatozoon sp., including 32/87 (37%) pools (1 to 10 larvae) of Amblyomma larvae, 21/105 (20%) pools (1 to 5 nymphs) of Amblyomma sculptum nymphs, 9/43 (21%) pools (1 to 5 nymphs) of Amblyomma dubitatumnymphs, and 1/12 (8%) A. sculptum adult female. The partial 18S rRNA sequence from one coati's blood sample and one representative of each positive tick species randomly selected from each area for sequencing (1,000 bp) showed 100% identity with sequences of H. procyonis from GenBank previously detected in coatis. Regarding H. procyonis infection, no statistical differences were obtained when comparing males vs. females (p-value 0.67), immature animals vs. adults (p-value 0.31), rainy vs. dry season (p-value 0.51) and sampling location (p-value 0.42). No noticeable alteration in blood parameters or heath status was observed in parasite animals. H. procyonis circulates in a high prevalence in coatis from central-western Brazil. Parasitemia fluctuates among different coatis' recaptures and apparently the infection has no influence in coatis' hematological and clinical parameters.
Assuntos
Apicomplexa , Carnívoros , Eucoccidiida , Procyonidae , Carrapatos , Animais , Apicomplexa/genética , Brasil/epidemiologia , Eucoccidiida/genética , Feminino , Masculino , Parasitemia/epidemiologia , Parasitemia/veterinária , Procyonidae/parasitologia , Carrapatos/parasitologiaRESUMO
Hepatozoon spp. are Apicomplexan protozoa that parasitize a wide diversity of vertebrate hosts. In Brazil, few studies have reported the occurrence of Hepatozoon spp. in rodent species. Additionally, an evaluation of the population structure and distribution of Hepatozoon species over several Brazilian biomes has not yet been performed. The present work aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from 31 genera sampled in five Brazilian biomes. Samples were submitted to PCR assays for Hepatozoon spp. targeting two regions of the 18S rRNA gene. Infection by Hepatozoon spp. was detected in 195 (42.2%) rodents comprising 24 genera. Phylogenetic analyses of 18S rRNA sequences grouped all sequences in the clade of Hepatozoon spp. previously detected in rodents and reptiles, apart from those detected in domestic/wild carnivores. These data raise two non-exclusive hypotheses: (i) rodents play an important role as intermediate or paratenic hosts for Hepatozoon infections in reptiles; and (ii) rodents do not seem to participate in the epidemiology of Hepatozoon infections of domestic/wild canids and felids in Brazil. TCS analyses performed with available 18S rRNA Hepatozoon sequences detected in rodents from Brazil showed the occurrence of six haplotypes, which were distributed in two large groups: one from rodents inhabiting the coastal region of Brazil and Mato Grosso state, and another from rodents from the central region of the country. A wide survey of the South American territory will help to elucidate the evolutionary history of Hepatozoon spp. parasitizing Rodentia in the American continent.
Assuntos
Apicomplexa/genética , Variação Genética , Roedores/parasitologia , Animais , Apicomplexa/patogenicidade , Brasil , Carnívoros/parasitologia , Haplótipos , Filogenia , Infecções Protozoárias em Animais/parasitologia , RNA Ribossômico 18SRESUMO
This study aimed to molecularly survey Bartonella in dogs from Chile. Quantitative real-time PCR (qPCR) for Bartonella spp. based on nuoG gene was performed in 139 blood samples taken from dogs belonging to rural localities of the Valdivia Province, Los Ríos region, southern Chile. nuoG qPCR-positive samples were submitted to conventional PCR assays for ftsZ, gltA, rpoB and nuoG genes and sequencing for speciation and phylogenetic analysis. Based upon qPCR results, Bartonella spp. occurrence in dogs was 4.3% (6/139). Out of six nuoG qPCR-positive samples, six, three, two and none showed positive results in cPCR assays based on gltA, ftsZ, rpoB and nuoG genes, respectively. Consistent sequencing results were obtained only for the ftsZ gene from sample #1532 (GeneBank accession number: MG252491), and gltA gene from samples #1535 (MG252490) and #1532 (148 bp fragment that was not deposited in GenBank). Phylogenetic analysis of ftsZ and gltA genes allowed speciation of two nuoG-positive samples, one as Bartonella vinsonii subsp. berkhoffii and the other as B. henselae. Bartonella vinsonii subsp. berkhoffii and B. henselae are detected for the first time in dogs from Chile, highlighting the importance of the canine population as a source of zoonotic agents and potential infection risk to humans.
Assuntos
Infecções por Bartonella/veterinária , Bartonella/classificação , Doenças do Cão/microbiologia , Animais , Bartonella/genética , Bartonella/isolamento & purificação , Infecções por Bartonella/diagnóstico , Infecções por Bartonella/epidemiologia , Infecções por Bartonella/microbiologia , Bartonella henselae/genética , Bartonella henselae/isolamento & purificação , Chile/epidemiologia , DNA Bacteriano/análise , Doenças do Cão/diagnóstico , Doenças do Cão/epidemiologia , Cães , FilogeniaRESUMO
Anaplasmataceae agents comprise obligate intracellular bacteria that can cause disease in humans and animals. Between August 2013 and March 2015, 31 Nasua nasua (coati), 78 Cerdocyon thous (crab-eating fox), seven Leopardus pardalis (ocelot), 110 wild rodents, 30 marsupials, and 42 dogs were sampled in the Pantanal wetland, Brazil. In addition, ectoparasites found parasitizing the animals were collected and identified. The present work aimed to investigate the occurrence of Anaplasmataceae agents in wild mammals, domestic dogs and ectoparasites, by molecular and serological techniques. Overall, 14 (17·9%) C. thous, seven (16·6%) dogs and one (3·2%) N. nasua were seroreactive to Ehrlichia canis. Nine dogs, two C. thous, one N. nasua, eight wild rodents, five marsupials, eight Amblyomma sculptum, four Amblyomma parvum, 13 A. sculptum nymphal pools, two Amblyomma larvae pools and one Polygenis (Polygenis) bohlsi bohlsi flea pool were positive for Ehrlichia spp. closely related to E. canis. Seven N. nasua, two dogs, one C. thous, one L. pardalis, four wild rodents, three marsupials, 15 A. sculptum, two Amblyomma ovale, two A. parvum and one Amblyomma spp. larval pools were positive for Anaplasma spp. closely related to A. phagocytophilum or A. bovis. The present study provided evidence that wild animals from Brazilian Pantanal are exposed to Anaplasmataceae agents.
Assuntos
Infecções por Anaplasmataceae , Anaplasmataceae , Animais Selvagens/microbiologia , Sifonápteros/microbiologia , Carrapatos/microbiologia , Anaplasmataceae/classificação , Anaplasmataceae/genética , Anaplasmataceae/imunologia , Anaplasmataceae/isolamento & purificação , Infecções por Anaplasmataceae/epidemiologia , Infecções por Anaplasmataceae/microbiologia , Infecções por Anaplasmataceae/veterinária , Animais , Animais Selvagens/imunologia , Anticorpos Antibacterianos/sangue , Brasil/epidemiologia , Cães/imunologia , Cães/microbiologia , Raposas/microbiologiaRESUMO
The order Chiroptera is considered the second largest group of mammals in the world, hosting important zoonotic virus and bacteria. Bartonella and hemotropic mycoplasmas are bacteria that parasite different mammals' species, including humans, causing different clinical manifestations. The present work aimed investigating the occurrence and assessing the phylogenetic positioning of Bartonella spp. and Mycoplasma spp. in neotropical bats sampled from Brazil. Between December 2015 and April 2016, 325 blood and/or tissues samples were collected from 162 bats comprising 19 different species sampled in five states of Brazil. Out of 322 bat samples collected, while 17 (5·28%) were positive to quantitative PCR for Bartonella spp. based on nuoG gene, 45 samples (13·97%) were positive to cPCR assays for hemoplasmas based on 16S rRNA gene. While seven sequences were obtained for Bartonella (nuoG) (n = 3), gltA (n = 2), rpoB (n = 1), ftsZ (n = 1), five 16S rRNA sequences were obtained for hemoplasmas. In the phylogenetic analysis, the Bartonella sequences clustered with Bartonella genotypes detected in bats sampled in Latin America countries. All five hemoplasmas sequences clustered together as a monophyletic group by Maximum Likelihood and Bayesian Inference analyses. The present work showed the first evidence of circulation of Bartonella spp. and hemoplasmas among bats in Brazil.
Assuntos
Infecções por Bartonella/veterinária , Bartonella/genética , Quirópteros , Infecções por Mycoplasma/veterinária , Mycoplasma/genética , Animais , Proteínas de Bactérias/genética , Bartonella/isolamento & purificação , Infecções por Bartonella/epidemiologia , Infecções por Bartonella/microbiologia , Brasil/epidemiologia , DNA Bacteriano/genética , Mycoplasma/isolamento & purificação , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Filogenia , Análise de Sequência de DNA/veterináriaRESUMO
Dogs and cats are often infected with vector-borne pathogens and play a crucial role as reservoirs and hosts in their life cycles. The aim of the present study was to investigate the occurrence of vector-borne pathogens among dogs and cats in the northwestern region of Rio Grande do Sul (RS) State, Brazil. One hundred and ten blood samples were collected from dogs (n=80) and cats (n=30). Laboratory analysis were carried out through stained blood smears, indirect enzyme-linked immunosorbent assay (ELISA) for Babesia vogeli and Ehrlichia canis (only for dogs) and polymerase chain reaction (PCR) aiming the detection of pathogens. The following pathogens were screened by PCR among dogs and cats: Babesia spp. and Hepatozoon spp. (18S rRNA gene), Anaplasma spp. (16S rRNA gene), and Ehrlichia spp. (dsb gene for dogs and 16S rRNA gene for cats) and Bartonella spp. (nuoG gene only for cats). Using blood smears structures morphologically compatible with piroplasms were found in 5.45% (6/110) of the samples. Anti-B. vogeli and anti-E. canis antibodies were detected in 91% (73/80) and 9% (7/80) of the dogs, respectively. All the seropositive dogs to E. canis were also to B. vogeli. Nineteen (17.3%) animals were positive to hemoparasites by PCR. After sequencing Rangelia vitalii 6/80 (7.5%), B. vogeli 3/80 (4%), Hepatozoon spp. 1/80 (1%), and Anaplasma spp. 1/80 (1%) were found in the dogs, and B. vogeli 2/30 (7%) and Bartonella spp. 6/30 (20%) were detected in the screened cats. No sample was positive for genes dsb and 16S rRNA of Ehrlichia spp. Only those animals which were positive for R. vitalii showed findings compatible with rangeliosis, such as anemia (100%), thrombocytopenia (67%), jaundice (50%), external bleeding (50%), and anorexia (50%). This is the first time that B. vogeli detected among cats in Southern Brazil.