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Piroplasmoses are one of the most prevalent arthropod-borne diseases of animals. The present work aimed to investigate the occurrence of piroplasmid in wild mammals, domestic dogs and ectoparasites in southern Pantanal region, central-western Brazil. For that purpose, blood or tissue samples from 31 Nasua nasua, 78 Cerdocyon thous, 7 Leopardus pardalis, 42 dogs, 110 wild rodents, and 30 marsupials, and 1582 ticks were submitted to PCR assays for piroplasmid targeting 18SrRNA and hps70 genes. Seven dogs, one C. thous,five L. pardalis, three N. nasua, six wild rodents, eight Amblyomma parvum, two Amblyomma sculptumand one Amblyomma ovalewere positive for piroplasmid-PCR assays. Genotypes closely related to Babesia vogeliwere detected in six dogs and five wild rodents. While genotypes closely related to Babesia caballiwere detected in one C. thous, one dog, one A. ovaleand one A. sculptum,genotypes closely related to Babesia bigeminaand Babesia boviswere detected in four A. parvumticks. Four sequences obtained from A. parvum, three coatis and one wild rodent were closely related to Theileria equi. Cytauxzoonspp. was detected in four ocelots. The present study revealed that wild and domestic animals in Brazilian southern Pantanal are exposed to different piroplasmid species.

The genus Rickettsiacomprises obligatory intracellular bacteria, well known to cause zoonotic diseases around the world. The present work aimed to investigate the occurrence of Rickettsiaspp. 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 parkeriand 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, ompBand htrAgenes, 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 sanguineussensu 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 Rickettsiaspecies. For 17 (40.4%) dogs, 33 (42.3%) crab-eating foxes, and two (33.3%) ocelots, homologous reactions to R. amblyommatisor 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 Rickettsiaspp. Among SFG Rickettsia-positive ticks samples, 93 (80.2%) belonged to A. parvum, 14 (12%) belonged to A. sculptumspecies, three (2.5%) belonged to A. auricularim, and six (5.2%) were Amblyommalarval pools. Thirty samples out of 117 qPCR positive samples for SFG Rickettsiaspp. also showed positivity in cPCR assays based on gltA, htrAand/or ompBgenes. The Blast analyses showed 100% identity with ‘CandidatusRickettsia andeanae’ in all 30 sequences obtained from gltA, htrAand/or ompBgenes. The concatenated phylogenetic analysis based on gltAand 17-kDa htrAgenes grouped the Rickettsiasequences obtained from tick samples in the same clade of ‘CandidatusRickettsia 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.

New genotypes of Anaplasmataceae agents have been detected in wild carnivores, birds and deer in Brazil. The present work aimed to investigate the presence of Ehrlichiaand Anaplasmaspecies in rodents sampled in Brazil. Additionally, a newly designed quantitative 5′ nuclease real-time multiplex PCR for Ehrlichiaand Anaplasmaspp. detection based on groELgene amplification was designed, showing high specificity and sensitivity (10 groELfragment copy/μL). Between 2000 and 2011, different rodent species [n=60] were trapped in 5 Brazilian biomes. Among 458 rodent spleen samples, 0.4% (2/458) and 2.4% (11/458) were positive for Ehrlichiaand Anaplasmaspp., respectively. Of 458 samples, 2.0% (9/458) and 1.1% (5/458) were positive for Anaplasmasp. and Ehrlichiasp., respectively, using conventional 16S rRNA PCR assays. Maximum Likelihood phylogenetic analyse based on a small region of 16S rRNA genes positioned the Anaplasmagenotypes in rodents near Anaplasma phagocytophilumor Anaplasma marginaleand Anaplasma odocoileiisolates. Ehrlichiagenotypes were closely related to E. canis. There was a low occurrence of Anaplasmaand Ehrlichiain wild and synanthropic rodents in Brazil, suggesting the circulation of new genotypes of these agents in rodents in the studied areas.

Anaplasmataceae agents, piroplasmids and Hepatozoonspp. have emerged as important pathogens among domestic and wild felines. The present work aimed to detect the presence of species belonging to the Anaplasmataceae family, piroplasmas and Hepatozoonspp. DNA in blood samples of domesticated and stray cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil. Between January and April 2013, whole blood samples were collected from 151 cats (54 males, 95 females and two without gender registration) in the city of Campo Grande, state of Mato Grosso do Sul, Brazil. DNA extracted from cat blood samples was submitted to conventional PCR assays for Theileria/Babesia/Cytauxzoonspp. (18S rRNA, ITS-1), Ehrlichiaspp. (16S rRNA, dsb, groESL), Anaplasmaspp. (16S rRNA, groESL) and Hepatozoonspp. (18S rRNA) followed by phylogenetic reconstructions. Out of 151 sampled cats, 13 (8.5%) were positive for Ehrlichiaspp. closely related to Ehrlichia canis, 1 (0.66%) for Hepatozoonspp. closely related to Hepatozoon americanumand Hepatozoonspp. isolate from a wild felid, 1 (0.66%) for Cytauxzoonsp. closely related do Cytauxzoon felis, and 18 (11.9%) for Babesia/Theileria(one sequence was closely related to Babesia bigemina, eight for Babesia vogeli, five to Theileriaspp. from ruminants [Theileria ovis, Theileria lestoquardi] and four to Theileriasp. recently detected in a cat). The present study showed that Ehrlichiaspp., piroplasmids (B. vogeli, Theileriaspp. and Cytauxzoonspp.) and, more rarely, Hepatozoonspp. circulate among stray and domesticated cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil.

Recently, tick and flea-borne pathogens have been detected in wild carnivores maintained in captivity in Brazilian zoos. Since free-roaming cats are frequently found in Brazilian zoos, they could act as reservoirs for arthropod-borne pathogens, which could be transmitted to endangered wild carnivores maintained in captivity in these institutions. On the other hand, stray cats in zoos may play a role as sentinels to pathogens that circulate among wild animals in captivity. The present work aimed to detect the presence of Anaplasmataceae agents, hemoplasmas, Bartonellaspecies, piroplasmas, and Hepatozoonsp. DNA in blood samples of 37 free-roaming cats in a Brazilian zoo. Three (8%) cats were positive for Anaplasmaspp. closed related to Anaplasma phagocytophilum; 12 (32%) cats were positive for hemoplasmas [two (5%) for Mycoplasma haemofelis, five (13.5%) for CandidatusMycoplasma haemominutum, and five (13.5%) for CandidatusMycoplasma turicensis]; 11 (30%) were positive for Bartonellaspp., six (16%) were positive Babesiavogeli and one (3%) for Theileriasp. Coinfection with multiple arthropod-borne agentes was observed in sampled cats. None of sampled cats were positive for Ehrlichiaspp., Cytauxzoonspp., or Hepatozoonspp. in PCR. This is the first molecular detection of Babesia vogeliand Theileriasp. in domestic cats in Brazil. The control of the population of free-roaming cats in these conservation institutions is much needed aiming to prevent the potential transmission to endangered wild animals maintained in captivity, such as wild neotropical wild felids, as well as to human beings visiting zoos.