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The species in the genus Neotrichodectes (Phthiraptera: Ischnocera) infest carnivores. Neotrichodectes (Nasuicola) pallidus (Piaget, 1880), which has been primarily found parasitizing Procyonidae mammals, has been recorded in ring-tailed coatis (Nasua nasua) in the Brazilian states of Minas Gerais, Pernambuco, Santa Catarina, Rio Grande do Sul and Pernambuco. We report a new record of N. pallidus in coatis in the state of Mato Grosso do Sul, central-western Brazil, using morphological (Light and Scanning Electronic Microscopy) and molecular approaches (PCR, sequencing and phylogenetic analysis). Coatis were sampled in two peri-urban areas of Campo Grande city, Mato Grosso do Sul state, Brazil, between March 2018 and March 2019, as well as in November 2021. Lice were collected and examined under light and Scanning Electron Microscopy. DNA was also extracted from nymphs and adults and submitted to PCR assays based on the 18S rRNA and cox-1 genes for molecular characterization. One hundred and one coatis were sampled from 2018 to 2019 and 20 coatis in 2021 [when the intensity of infestation (II) was not accessed]. Twenty-six coatis (26/101-25.7%) were infested with at least one louse, with a total of 59 lice collected in 2018-2019. The II ranged from one to seven lice (mean 2.2 ± SD 1.7). The louse species was confirmed based on the following morphological characteristics: female gonapophyses rounded with the setae along anterior region but not in the medial margin; the male genitalia with a parameral arch not extending beyond the endometrial plate. The same ornamentation was observed on the abdomen of the females, males, and nymphs. The nymphs and the eggs were described in detail for the first time. The obtained 18S rRNA and cox1 sequences from N. pallidus clustered in a clade with other sequences of Ischnocera species. In the present study, a new record of the louse N. pallidus in central-western Brazil was provided, along with new insights into the morphological features of this species, with the first morphology contribution of nymphal and eggs stages., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

White-eared opossums (Didelphis albiventris) are well adapted to anthropized areas. The increased contact with domestic animals and humans mediates the transmission of arthropod-borne pathogens. Despite the worldwide occurrence of tick-borne Anaplasmataceae and Hepatozoidae species in a variety of vertebrates, few studies reported serological evidence or molecular detection of theses agentes in marsupials. Up to now, while Ehrlichia/Anaplasma spp. have only been detected in marsupials from Brazil, Hepatozoon spp. have been reported in marsupials from Chile, Australia and Brazil. The present work aimed to investigate, using molecular techniques and blood smear analysis, the presence of Ehrlichia spp., Anaplasma spp., and Hepatozoon sp. in the blood and ticks collected from D. albiventris in urban forest fragments from midwestern Brazil. Between May and December 2017, 43 D. albiventris (27 males and 16 females) were captured for blood and tick collection in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil. Ticks (46 Amblyomma dubitatum nymphs and 24 Amblyomma spp. larvae) were collected from 14 out 43 (32.5%) of the white-eared opossums. Panoptic-stained blood smears were performed using peripheral blood (tail tip) of the captured opossums. DNA extracted from blood and tick samples were subjected to PCR/qPCR assays for Anaplasmataceae agents (rrs, gltA, groEL, sodB, and dsb genes, and 23S-5S intergenic region) and Hepatozoon spp. (18S rRNA gene), followed by Sanger sequencing, BLASTn and phylogenetic analyses. An inclusion resembling Ehrlichia morulae was found in a white-eared opossum's monocyte from a blood smear stained with Panoptic. Five (11.63% [5/43]) white-eared opossums' blood samples and 7 (25% [7/28]) tick samples (2 pools of Amblyomma spp. larvae and 5 pools of A. dubitatum nymphs) were positive for Anaplasmataceae via a PCR assay targeting the conserved rrs gene. Phylogenetic analysis based on the rrs gene positioned three sequences obtained from opossums and ticks together as a subclade within the Ehrlichia canis clade. However, all samples were negative in a qPCR assay specific for E. canis based on the dsb gene. Phylogenetic analyses positioned the gltA and 23S-5S ITS sequences obtained from opossums' blood samples in a separate clade from the other validated Ehrlichia species. One (2.3% [1/43]) opossum blood sample was positive for the 18S rRNA gene of Hepatozoon sp. The phylogenetic analysis positioned the Hepatozoon sp. sequence obtained from a D. albiventris specimen in a clade with a sequence previously detected in a black storm petrel (Oceanodroma melania) from Mexico. All the other sequences of Hepatozoon sp. previously detected in marsupials from Brazil were positioned in a separated clade. The present work showed the occurrence of putative novel genotypes of Ehrlichia sp. and Hepatozoon sp. in white-eared opossums and associated A. dubitatum ticks from midwestern Brazil., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

It has been estimated that 75% of emerging infectious diseases comprise zoonoses, whose majority have free-living animals as reservoirs and are mainly transmitted by arthropod vectors. Although rodents represent important Bartonella reservoirs, there are few studies on the genotypic characterization of Bartonella species commonly found in this taxon and from different Brazilian biomes. Therefore, the present study aimed to investigate the occurrence, isolate and molecularly, morphologically and phenotypically characterize a new Bartonella species infecting free-living rodents sampled in the Brazilian Pantanal, the largest wetland in South America. For this purpose, 129 free-living rodents (79 Thrichomys fosteri, 4 Clyomys laticeps, and Oecomys mamorae) were captured. While blood samples were collected from 57 T. fosteri, 4 C. laticeps and 32 O. mamorae; spleen samples were collected from 22 T. fosteri and 14 O. mamorae. Blood and spleen samples were submitted to a qPCR for Bartonella spp. targeting the nuoG gene, using DNA samples extracted directly from blood/spleen, after passage in pre-enrichment liquid culture, and from colonies obtained from solid culture on chocolate agar. Combining all techniques, occurrence of 24.8% for Bartonella sp. was found among the sampled rodents. One Bartonella isolate (strain 56A) obtained from a T. fosteri's blood sample was closely related to the Bartonella vinsonii complex and selected for Whole Genome Sequencing (WGS) hybrid approach using Illumina NovaSeq and Nanopore sequencing platforms. This strain exhibits a circular 2.7 Mbp genome with an average C+G content of 39% and encoding to 2239 genes. In the phylogenomics based on 291 shared protein-coding genes, this strain was positioned in a unique clade, closely related to Bartonella vinsonii subsp. vinsonii, B. vinsonii subsp. berkhoffii and B. visonii subsp. arupensis. An Average Nucleotide Identity of 85% was found between the obtained isolate and Bartonella species belonging to B. vinsonii complex. These findings supported the separation of this strain, now formally named as Bartonella machadoae sp. nov., from the Bartonella vinsonii complex. In addition, Bartonella machadoae sp. nov. was characterized by capnophilic, microaerophilic and aerobic small rods with absence of pili and flagella. Phylogenetic and distance analyses based on five concatenated molecular markers suggest that Bartonella machadoae may parasite rodents from different Brazilian biomes. In conclusion, we described biochemical, phenotypic and genomic characteristics of Bartonella machadoae nov. sp. isolated from blood samples of T. fosteri rodents from the Brazilian Pantanal., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Hemoplasmas have already been detected in bats in the United States of America, Spain, Australia, Chile, Brazil, Peru, Belize, Nigeria, Costa Rica, Germany, Switzerland and New Caledonia. The recent detection of hemoplasmas closely related to Mycoplasma haematohominis, an agent causing disease in humans, emphasizes the need for additional studies on the diversity of hemoplasmas in bats. The present work aimed to investigate the occurrence and assess the phylogenetic positioning and genetic diversity of hemoplasmas in bats and associated ectoparasites sampled in central-western Brazil. Overall, 43% (58/135) sampled bats and 1.56% (1/64) bat flies (Megistopoda aranea) were positive for hemoplasmas, however, twenty-four and two hemoplasma sequences were obtained from PCR assays targeting 16S and 23S rRNA genes, respectively, since the majority of the obtained amplicons showed faint bands in agarose gel electrophoresis. The obtained 16S rRNA sequences showed to be broadly distributed along the phylogenetic tree, albeit positioned within the 'Haemofelis group' and clustering with other bat-associated hemoplasmas. Twelve 16S rRNA hemoplasma genotypes were found among the 24 obtained sequences. When compared to other bat-related hemoplasmas sequences retrieved from the Genbank, 52 genotypes were found. The two 23S rRNA sequences obtained were positioned as a sister clade to "Candidatus Mycoplasma haematohydrochaerus", M. haemofelis and M. haemocanis. High genetic diversity was found among 16S rRNA hemoplasma sequences detected in non-hematophagous bats from central-western Brazil and previously detected in other regions of the world. Even though the genotype analysis showed that hemoplasmas from the same genus tend to group together, the results from the unipartite and bipartite analyses did not robustly support the hypothesis. Further studies addressing the specificity of hemoplasma genotypes according to bat species and genera should be performed., (Copyright © 2021. Published by Elsevier B.V.)

Worldwide, Bartonella species are known to infect a wide range of mammalian and arthropod hosts, including humans. The current study aimed to investigate the prevalence of Bartonella spp. in synanthropic mammals captured in peri-urban areas from Central-Western and Southern Brazil and their ectoparasites. For this aim, 160 mammals belonging to four species, and 218 associated arthropods were sampled. DNA was extracted and subjected to different Bartonella screening assays. Additionally, blood samples from 48 small rodents were submitted to liquid BAPGM culture followed by qPCR assay and solid culture. Two out of 55 Rattus captured in Santa Catarina state were PCR-positive for Bartonella when targeting the nuoG, 16S, and ITS loci. Sequences showed high homology with Bartonella coopersplainsensis. Conversely, all 48 small rodents, 14 capybaras and 43 opossum DNA samples from animals trapped in Mato Grosso do Sul were Bartonella negative in the HRM real time PCR assays targeting the ITS locus and gltA gene. Additionally, all mammal-associated ectoparasites showed negativity results based on HRM real time PCR assays. The present study showed, for the first time, the occurrence of B. coopersplainsensis in Brazil, shedding some light on the distribution of rats-related Bartonella in South America. In addition, the majority of rodents and marsupials were negative for Bartonella spp. Since B. coopersplainsensis reservoirs - Rattus spp. - are widely dispersed around the globe, their zoonotic potential should be further investigated., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The Anaplasmataceae family is composed of obligatory intracellular Gram-negative bacteria transmitted by arthropod vectors. In Brazil, with the exception of Anaplasma marginale, little is known about the occurrence of other Anaplasma and Ehrlichia species infecting cattle. The present study aimed at investigating the occurrence of Anaplasma spp. and Ehrlichia spp. in beef cattle (Bos indicus) sampled in the Brazilian Pantanal, an area prone to periodic flooding and endemic for bovine trypanosomiasis. Blood samples from 400 cattle were collected and screened by PCR assays based on rrs and dsb genes from Anaplasma spp. and Ehrlichia spp., respectively. Positive samples for Anaplasma spp. were subjected to qPCR assays based on the msp-2 gene and nPCR based on the groEL gene. As a result, 4.75% (19/400) and 48.12% (167/347) were positive for Anaplasma platys and Ehrlichia minasensis, respectively. Besides, positivity of 56.75% (227/400) for A. marginale and seropositivity of 90.75% (363/400) for Trypanosoma vivax were found. A high rate of co-infection was observed (67.25%), from which the co-infection by A. marginale and E. minasensis was more frequently found in calves than cows. Interestingly, none of the animals presenting co-infection showed anemia or other clinical signs. The present study showed, for the first time, the occurrence of A. platys and E. minasensis in beef cattle in the southern Pantanal, as well as a high rate of co-infection by A. marginale, E. minasensis and T. vivax in the sampled animals., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Even though Hepatozoon spp. has been molecularly detected in several wild animals in Brazil, there is no report on the occurrence of Hepatozoon spp. DNA in bats in Brazil. This study aimed at detecting Hepatozoon, in addition to ectoparasites, in non-hematophagous bats sampled in central-western Brazil using blood smears, hematoxylin-eosin (HE)-staining liver/spleen preparations and molecular and phylogenetic techniques. A total of 135 spleen, 127 liver, and 133 blood samples were collected from 135 non-hematophagous bats from 12 different species in two different sites in Campo Grande, Mato Grosso do Sul state, in the Brazilian Cerrado region. Spleen and blood DNA samples were submitted to two conventional PCR protocols for Hepatozoon spp. based on 18S rRNA. No Hepatozoon spp. gamonts or meronts were observed in blood smears and HE-stained-liver preparations, respectively. While none of the spleen samples was positive for Hepatozoon spp. in the PCR assays, 5 (3 %) blood samples contained 18S rRNA Hepatozoon DNA, including 2/37 (5 %) Artibeus lituratus, 2/32 (6 %) A. planirostris, and 1/23 (4 %) Platyrrhinus lineatus. Out of 5 bats positive for Hepatozoon spp., 3 were parasitized by either Macronyssidae/Spinturnicidae mites or Streblidae flies. BLAST analysis showed that the sequences detected in bats had >99 % identity with Hepatozoon sequences detected in amphibians and reptiles from Brazil, including Hepatozoon caimani detected in Caiman crocodilus. The phylogenetic inferences estimated by the Maximum Likelihood and Bayesian methods clustered the Hepatozoon sequences detected in Brazilian bats with those detected in reptiles and amphibians., Competing Interests: Declaration of Competing Interest None, (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Globally, hemotropic mycoplasmas (hemoplasmas) comprise an emerging or remerging bacteria group that attaches to red blood cells of several mammal's species and in some cases, causing hemolytic anemia. Herein, we assessed the occurrence, genetic diversity, the factors coupled to mammals infection, and the phylogeographic distribution of hemoplasmas in sylvatic and synanthropic mammals and their associated ectoparasites from Brazil. We collected spleen and/or blood samples from synanthropic rodents (Rattus rattus [N = 39] and Mus musculus [N = 9]), sylvatic rodents (Hydrochoerus hydrochaeris [N = 14]) and opossums (Didelphis albiventris [N = 43]). In addition, ticks (Amblyomma spp. [N = 270] and lice (Polyplax spinulosa [N = 6]) specimens were also sampled. Using a PCR targeting the 16S rRNA region, out of 48 small rodents, 14 capybaras and 43 opossums DNA samples, hemoplasma DNA was found in 25%, 50%, and 32.5% animals, respectively. Besides, we reported hemoplasma DNA in Amblyomma sp. (22.2% [2/9]) and lice (100% [2/2]) pools samples from rats, and one female A. sculptum DNA sample (3% [1/33]) obtained from a capybara. Additionally, and in agreement with ML analysis, the network analyses showed a clear phylogenetic separation among the hemoplasmas genotypes found in the different host species sampled, thus, suggesting the absence of cross-species hemoplasmas transmission between the mammals trapped. Finally, using the NTC network analysis, we reported the same 16S rRNA Mycoplasma genotype circulating in Rattus sampled in Brazil, Hungary, and Japan., Competing Interests: Declaration of competing interest None., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Distinct species of Trypanosoma have been documented sharing the same hosts in different environments in intricate transmission networks. Knowing this, this study investigated the role of different hosts in the transmission cycles of Trypanosoma species in the Pantanal biome. The mammals were sampled from November 2015 to October 2016. We sampled a total of 272 wild mammals from 27 species belonging to six orders and 15 families, and three species of triatomines (n = 7). We found high parasitemias by Hemoculture test for Trypanosoma cruzi (TcI), Trypanosoma rangeli, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, and high parasitemias by Microhematocrit Centrifuge Technique for Trypanosoma evansi. The carnivore Nasua nasua is a key host in the transmission cycles since it displayed high parasitemias for T. cruzi, T. evansi and T. rangeli. This is the first report of high parasitemias in Tamandua tetradactyla and cryptic infection in Dasypus novemcinctus by T. cruzi; cryptic infection by T. evansi in Eira barbara, Euphractus sexcinctus and Dasyprocta azarae. The collection of Panstrongylus geniculatus increased the geographic distribution of this vector species in the South America. Our results indicate that Trypanosoma species circulate in a complex reservoir system including different host species with different infective competences., (Copyright © 2019 Elsevier B.V. All rights reserved.)

There are few studies on the genetic diversity of Anaplasma marginale in Brazilian cattle herds, especially about beef cattle. The objective of this study was to evaluate the genetic diversity of A. marginale, based on the msp1α gene in Bos taurus indicus sampled from the Brazilian Pantanal. Aliquots of blood with and without EDTA were taken from 400 cattle (200 cows and 200 calves) across five extensive farms. The samples were submitted to the indirect immunoenzymatic assay (iELISA), quantitative real-time PCR (qPCR) for the msp1β gene and to the semi-nested (sn) PCR for the msp1α gene. Positive samples were sequenced by the Sanger method and subjected to diversity analysis using the RepeatAnalyser software. The percentage of positive animals by iELISA, qPCR and (sn) PCR was 72.2% (289/400), 56.7% (227/400) and 23% (52/227), respectively. Cows (154/200) showed to be significantly more seropositive than calves (135/200). In qPCR, the number of calves and average quantification value (138/200; 1.3 × 10 6 ) A. marginale msp1α copies per μL proved to be higher when compared to that found for the cows (89/200; 3.9 × 10 4 ). The microsatellite analysis of the 26 sequences obtained from the msp1α gene revealed the presence of E (77%), C (15.4%) and B (7.7%) genotypes. Fourteen A. marginale strains were identified in the studied region, with eight that have never before been described in the literature (τ-10-13-13-18; τ-27-18; EV8-EV8-17; α-β-β-β-100; EV7-11-10-15; τ-11-11-27-18; τ-11-10-15; τ-27-13-18). Beef cattle are highly exposed to A. marginale in the Brazilian Pantanal. Moreover, a high genetic diversity of A. marginale, with eight new strains, was found in the studied region. While cows may act as chronic carriers, perpetuating the pathogen within the herd, male beef calves sold to other regions may disperse these strains., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

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 sculptum and one Amblyomma ovale were positive for piroplasmid-PCR assays. Genotypes closely related to Babesia vogeli were detected in six dogs and five wild rodents. While genotypes closely related to Babesia caballi were detected in one C. thous, one dog, one A. ovale and one A. sculptum, genotypes closely related to Babesia bigemina and Babesia bovis were detected in four A. parvum ticks. Four sequences obtained from A. parvum, three coatis and one wild rodent were closely related to Theileria equi. Cytauxzoon spp. was detected in four ocelots. The present study revealed that wild and domestic animals in Brazilian southern Pantanal are exposed to different piroplasmid species., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

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., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

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., (Copyright © 2017 Elsevier B.V. All rights reserved.)

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., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Anaplasmataceae agents, piroplasmids and Hepatozoon spp. 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 Hepatozoon spp. 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/Cytauxzoon spp. (18S rRNA, ITS-1), Ehrlichia spp. (16S rRNA, dsb, groESL), Anaplasma spp. (16S rRNA, groESL) and Hepatozoon spp. (18S rRNA) followed by phylogenetic reconstructions. Out of 151 sampled cats, 13 (8.5%) were positive for Ehrlichia spp. closely related to Ehrlichia canis, 1 (0.66%) for Hepatozoon spp. closely related to Hepatozoon americanum and Hepatozoon spp. isolate from a wild felid, 1 (0.66%) for Cytauxzoon sp. 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 Theileria spp. from ruminants [Theileria ovis, Theileria lestoquardi] and four to Theileria sp. recently detected in a cat). The present study showed that Ehrlichia spp., piroplasmids (B. vogeli, Theileria spp. and Cytauxzoon spp.) and, more rarely, Hepatozoon spp. circulate among stray and domesticated cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Two main genotypes in Trypanosoma cruzi subpopulations can be distinguished by PCR amplification of sequences from the mini-exon gene non-transcribed spacer, respectively, T. cruzi I (TCI) and T. cruzi II (TCII). This technique is also capable of distinguishing a third assemblage of subpopulations that do not fit in these genotypes and that remain known as zymodeme Z3 (Z3). The distribution pattern as well as the mammalian host range of this latter T. cruzi sublineage still remains unclear. Thus, the intention of our study was to increase the information regarding these aspects. The mini-exon analysis of T. cruzi isolates obtained from sylvatic animals in the Amazon Forest, Atlantic Rainforest, Caatinga and Pantanal showed that prevalence of the Z3 subpopulation in nature was low (15 out of 225 isolates, corresponding to 7%). A higher prevalence of Z3 was observed in the Caatinga (15%) and the Pantanal (12%). Infection by Z3 was observed in mammalian hosts included in Carnivora, Chiroptera, Didelphimorphia, Rodentia and Xernathra. The T. cruzi Z3 subpopulation was observed also in mixed infections (33%) with TCI (n=2) and TCII (n=3). These results demonstrate that T. cruzi Z3 displays a wider distribution and host range than formerly understood as it has been demonstrated to be able infect species included in five orders of mammalian host species dispersed through all forest strata of the four Brazilian biomes evaluated.

Few studies have been conducted to investigate the role played by the order Chiroptera in the sylvatic transmission cycle of Trypanosoma cruzi or their putative association with the main genotypes of the parasite. Here, the purpose was to enlarge the knowledge of this issue, in this sense, 93 specimens of bats included in 4 families, respectively Molossidae, Noctilionidae, Phyllostomidae and Vespertilionidae collected in distinct regions of Brazil were submitted to fresh blood smears and hemocultures. No patent parasitemia was observed but positive hemocultures by T. cruzi were observed in 14% (13/93) of examined samples. The majority of the parasite isolates were obtained from Phyllostomus hastatus (80%) captured in one same buriti hollow palm tree in the Cerrado region. Multilocus enzyme electrophoresis (MLEE) analyses showed that the genetic distance among these isolates was 0.35, almost the same observed when all the isolates (excluding the reference strains) were analyzed (0.40). No correlation of zymodeme with bat genera, species or geographic region of its origin could be observed, moreover, correlation of zymodeme and genotype of the parasite was not strict. Ten out of 14 T. cruzi isolates obtained from bats corresponded to the TCII genotype. Chiropterans with TCI, TCII/TCIII mixed infection as well as Trypanosoma rangeli in single or mixed infections were observed. These results show that bats may harbor and are probably important maintainers of the main genotypes (TCI, TCII, TCIII/Z3) of T. cruzi. These results support the absence of an association of TCII with any mammal order and show that bats, mainly P. hastatus, may act as amplifier hosts of TCII subpopulations of T. cruzi.