Health of Holochilus chacarius (Rodentia: Cricetidae) in rice agroecosystem in a neotropical wetland assessed by histopathology.

Small mammals have a short lifetime and are strictly associated with their environment. This work aimed to use histopathology to assess the health of Holochilus chacarius in a rice agroecosystem in the Pantanal of Mato Grosso do Sul. During necropsy, fragments of the lung, kidney, skin, liver, and reproductive system of 33 animals were collected and submitted to histological processing. Tissue damages were evaluated as mild, moderate, and severe and arranged in a matrix for further statistical analysis. Furthermore, we used generalized linear models to verify the influence of tissue changes on the body condition, obtained by a regression between body mass and length. In the lungs, we found an intense inflammatory infiltrate associated with anthracosis that had a negative influence on the body's condition. Also, we observed degenerative and inflammatory changes in the liver, kidneys, skin, and reproductive system that ranged from mild to moderate. The histopathological lesions observed in this study may be associated with environmental alterations of anthropic origin such as the exposure to soot from wildfires and heavy metals, evidenced by lesions in the lung, kidney, and liver. The present study provided a histopathological matrix as a new approach that allows to classify and quantify the tissue alterations. Tissue changes when associated with body condition demonstrated to be an effective tool to assess the health of small free-living mammals, showing that these animals can be used as bioindicators of environmental condition.

Exploring interactions between parasites and their hosts in the Pantanal floodplain using an ecological network approach.

The study of host-parasite interactions is essential to understand the role of each host species in the parasitic transmission cycles in a given community. The use of ecological network highlights the patterns of interactions between hosts and parasites, allowing us to evaluate the underlying structural features and epidemiological roles of different species within this context. Through network analysis, we aimed to understand the epidemiological roles of mammalian hosts species (n = 67) and their parasites (n = 257) in the Pantanal biome. Our analysis revealed a modular pattern within the network, characterized by 14 distinct modules, as well as nestedness patterns within these modules. Some key nodes, such as the multi-host parasites Trypanosoma cruzi and T. evansi, connect different modules and species. These central nodes showed us that various hosts species, including those with high local abundances, contribute to parasite maintenance. Ectoparasites, such as ticks and fleas, exhibit connections that reflect their roles as vectors of certain parasites. Overall, our findings contribute to a comprehensive understanding of the structure of host-parasite interactions in the Pantanal ecosystem, highlighting the importance of network analysis as a tool to identifying the main transmission routes and maintenance of parasites pathways. Such insights are valuable for parasitic disease control and prevention strategies and shed light on the broader complexities of ecological communities.

Trypanosomatid diversity in a bat community of an urban area in Campo Grande, Mato Grosso do Sul, Brazil.

Bats have a long evolutionary history with trypanosomatids, but the role of these flying mammals on parasite transmission cycles in urban areas, especially for Trypanosoma and Leishmania species, remains poorly known. The objective of this study was to evaluate the species richness of trypanosomatids parasitizing a bat community in Campo Grande (CG), a state capital within the Cerrado of the Brazilian Midwest. We evaluated 237 bats of 13 species by means of hemoculture and molecular detection in spleen samples. The bat community of CG appears to participate in the transmission cycles of various species of trypanosomatids. We report an overall trypanosomatid detection rate of 34.2% (n = 81), involving 11 out of 13 sampled bat species. We identified six species of trypanosomatids from 61 bats by analyzing SSU rRNA and/or kDNA: Trypanosoma cruzi DTU TcI, T. c. marinkellei, T. dionisii, Leishmania infantum, L. amazonensis, and T. janseni, with this latter being detected by hemoculture for the first time in a bat species. We also detected a Molecular Operational Taxonomic Unit, Trypanosoma sp. DID, in the phyllostomids Glossophaga soricina and Platyrrhinus lineatus. The highest trypanosomatid richness was observed for Sturnira lilium, which hosted three species: L. infantum, T. dionisii and T. janseni. Given that visceral leishmaniasis is endemic in CG, special focus should be placed on L. infantum. Moreover, L. amazonensis and T. cruzi warrant attention, since these are zoonotic parasites responsible for human cases of tegumentary leishmaniasis and Chagas disease, respectively. In this respect, we discuss how bat communities may influence the Leishmania spp. transmission in endemic areas.

The relationships among Leishmania infantum and phyllostomid bats assessed by histopathological and molecular assays.

Bats have been reported as reservoir host of Leishmania spp. worldwide, mostly by molecular detection. However, it is still unclear whether bats act as reservoirs of Leishmania infantum to sandflies vectors. In this sense, the investigation of amastigotes forms in the target organs, and the characterization of their associated inflammation, may help to clarify the epidemiological importance of bats in endemic areas for leishmaniasis. The aim of this work was to investigate the host-parasite relationships under microscopic evaluation and predict the epidemiological role of two phyllostomid bats species naturally infected by L. infantum in an endemic area for human leishmaniasis. Fragments of skin, liver and spleen of L. infantum positive and negative bats (Artibeus planirostris and Carollia perspicillata) by qPCR, were studied by histological and immunohistochemical techniques. Both groups, positive and negative, did not show differences in the histopathological study, presenting only discrete tissue changes. Liver and skin showed mild inflammatory reactions. Findings on spleen consisted of reactivity of the lymphoid follicles, expressive presence of apoptotic cells and macrophages containing abundant phagocytic cells debris. We did not find amastigote forms in tissues by histological and IHC techniques in positive qPCR bats. Our results allow us to hypothesize that phyllostomid bats seem to have an important role in reducing the risk of transmission, possibly acting as dead-end host.

Exploring the Hidden World of Vectors of Chagas Disease: A Fascinating Look at the Taxonomic Aspects of the Psammolestes Genus (Hemiptera, Triatominae).

Chagas disease (CD) is a neglected illness affecting approximately seven million individuals, with vector transmission occurring via triatomine bugs. The Rhodniini tribe comprises 24 species, grouped into the Rhodnius and Psammolestes genera. Given the importance of accurately identifying CD vectors, the taxonomy of Psammolestes spp. was revisited using morphological and morphometric data. Specimens of P. tertius, P. coreodes, and P. arthuri were collected, and the morphological characteristics of the head, thorax, abdomen, and eggs were analyzed. Morphometric studies of eggs were also conducted. Dichotomous keys allowing for the differentiation of Psammolestes spp. were elaborated based on adult insect and egg morphological characteristics. Through these studies, it was possible to differentiate the three Psammolestes species and confirm that this genus should not be classified under the Rhodnius genus, contributing to Rhodniini taxonomy.

Molecular Survey of Hemotropic Mycoplasma spp. and Bartonella spp. in Coatis (Nasua nasua) from Central-Western Brazil.

Even though previous works showed molecular evidence of hemotropic Mycoplasma spp. (hemoplasmas) in ring-tailed coatis (Nasua nasua) from Brazil, Bartonella sp. has not been reported in these mammals so far. The present study aimed to detect the above-mentioned agents in coatis' blood and associated ectoparasites, assessing the association between these infections and red blood parameters. Between March 2018 and January 2019, coati (n = 97) blood samples, Amblyomma sp. ticks (2242 individual ticks, resulting in 265 pools), and Neotrichodectes pallidus louse (n = 59) were collected in forested urban areas from midwestern Brazil. DNA extracted from coatis' blood, and ectoparasite samples were submitted to quantitative PCR (qPCR) (16S rRNA) and conventional PCR (cPCR) (16S rRNA and 23S rRNA) for hemoplasmas and qPCR (nuoG gene) and culturing (only blood) for Bartonella spp. Two different hemoplasma genotypes were detected in blood samples: 71% coatis positive for myc1 and 17% positive for myc2. While 10% of ticks were positive for hemoplasmas (myc1), no louse was positive. The estimated bacterial load of hemoplasmas showed no association with anemia indicators. All coatis were negative for Bartonella sp. in qPCR assay and culturing, albeit two Amblyomma sp. larvae pools, and 2 A. dubitatum nymph pools were positive in the qPCR. The present work showed a high occurrence of hemoplasmas, with two distinct hemoplasma genotypes, in coatis from forested urban areas in midwestern Brazil.

Corrigendum: Leishmania infantum infecting the carnivore Nasua nasua from urban forest fragments in an endemic area of visceral leishmaniasis in Brazilian Midwest.

[This corrects the article DOI: 10.3389/fcimb.2022.1050339.].

Molecular diversity of Sarcocystis spp. in opossums (Didelphis spp.) from Southeastern and Midwestern Brazil.

South American opossums (Didelphis spp.) are definitive hosts of Sarcocystis neurona, Sarcocystis speeri, Sarcocystis lindsayi and Sarcocystis falcatula. In Brazil, diverse studies have demonstrated a high frequency of Sarcocystis falcatula-like in sporocysts derived from opossums, and high genetic diversity has been observed in surface antigen-encoding genes (SAGs). In this study, genetic diversity of Sarcocystis spp. derived from Didelphis albiventris and Didelphis aurita from the cities of Campo Grande and São Paulo, was accessed by sequencing SAG2, SAG3, SAG4, the first internal transcribed spacer (ITS-1) and cytochrome c oxidase subunit I (cox1). Molecular identification was performed for 16 DNA samples obtained from sporocyst or culture-derived merozoites. The ITS-1, cox1, and SAG3 fragments were cloned, whereas SAG2 and SAG4 were sequenced directly from PCR products. Four alleles variants were found for SAG2, 13 for SAG3 and seven for SAG4, from which four, 13 and four, respectively, were novel. Twenty-seven allele variants were found for ITS-1, all phylogenetically related to S. falcatula-like previously described in Brazil. Sarcocystis sp. phylogenetically related to Sarcocystis rileyi was evidenced by cox1 in three opossums. Further studies are needed to clarify the role of Didelphis spp. as definitive hosts of Sarcocystis spp. other than that previous described.

Retrospective molecular investigation of Mayaro and Oropouche viruses at the human-animal interface in West-central Brazil, 2016-2018.

Mayaro virus (MAYV, Togaviridae) and Oropouche orthobunyavirus (OROV, Peribunyaviridae) are emerging enzootic arboviruses in Latin America. Outbreaks of febrile illness associated with MAYV and OROV have been reported among humans mainly in the northern region of Brazil since the 1980s, and recent data suggest these viruses have circulated also in more populated areas of western Brazil. MAYV shares mosquito vectors with yellow fever virus and it has been historically detected during yellow fever epidemics. Aiming to investigate the transmission of OROV and MAYV at the human-animal interface during a yellow fever, chikungunya and Zika outbreaks in Brazil, we conducted a retrospective molecular investigation in 810 wild and domestic animals, 106 febrile patients, and 22.931 vectors collected from 2016 to 2018 in Cuiaba and Campo Grande metropolitan regions, western Brazil. All samples tested negative for OROV and MAYV RNA by RT-qPCR. Findings presented here suggest no active circulation of MAYV and OROV in the sampled hosts. Active surveillance and retrospective investigations are instrumental approaches for the detection of cryptic and subclinical activity of enzootic arboviruses and together serve as a warning system to implement appropriate actions to prevent outbreaks.

Kinetoplastid Species Maintained by a Small Mammal Community in the Pantanal Biome.

Kinetoplastids include species economically important in agriculture, livestock, and human health. We evaluated the richness of kinetoplastids that infect small mammals in patches of unflooded forests in the Pantanal biome, an area where we hypothesize that its diversity is higher than currently recognized. Hemocultures (HC) and Next Generation Sequencing (NGS) targeting the 18S rDNA gene were employed for the detection of kinetoplastids. We grouped the positive samples into pools for each small mammal species (Monodelphis domestica, Thylamys macrurus, Oecomys mamorae, Thrichomys fosteri, Clyomys laticeps, and Holochilus chacarius). Eight parasite species were identified: Leishmania amazonensis, L. infantum; Trypanosoma cascavelli (HC + NGS), T. cruzi, T. lainsoni, T. rangeli (HC + NGS), Trypanosoma sp. DID, and Neobodo sp. The use of a tool as sensitive as NGS has increased our awareness of the diversity of kinetoplastids, as well as their host range, with emphasis on the species O. mamorae (seven kinetoplastid species, excepting T. cascavelli in a pool of nine individuals) and T. macrurus (four kinetoplastid species in a single individual). Furthermore, L. infantum and L. amazonensis infections were described in small mammals from this region for the first time. These findings make it mandatory to revisit the kinetoplastids/host associations proposed so far.

Brucellosis in the Brazilian Pantanal wetland: threat to animal production and wildlife conservation.

This review was performed to gather knowledge about brucellosis in livestock and wildlife in the Brazilian Pantanal, a biome with a huge biodiversity and extensive livestock production. Following the preferred reporting items for narrative review guidelines and using the terms "Brucella" and "Pantanal," we explored the PubMed, SciELO, Jstor, Science Direct, and Scholar Google databases. Information on host species, diagnostic test, number of positive animals, and positivity rates were acquired. Articles dating from 1998 to 2022 registered 14 studies including cattle, dogs, and the following wildlife species: Ozotoceros bezoarticus, Sus scrofa, Tayassu peccari, Nasua nasua, Cerdocyon thous, Panthera onca, Dasypus novemcintus, Cabassous unicinctus, Euphractus sexcinctus, Priodontes maximus, Myrmecophaga tridactyla and Hydrochoerus hydrochaeris. Brucella occurrence in cattle was demonstrated through the serological confirmatory test 2-mercaptoetanol. Molecular diagnosis detected Brucella abortus in dogs, smooth Brucella in O. beoarticus, and Brucella spp. in T. peccari. Cattle may have a pivotal importance in maintenance and spreading of Brucella spp. due to their high population density, environmental contamination from abortion of infected cows, and eventual excretion of B. abortus S19 strain from vaccinated heifers. The occurrence of Brucella spp. in O. bezoarticus and T. peccari represent a risk for wildlife conservation. These data indicate that Brucella spp. are enzootic in the Pantanal wetland, sustained by a reservoir system including domestic and wild mammals. Due to marked seasonality and large populations of wildlife species sharing same environments with livestock, brucellosis acquires great complexity in Pantanal and, therefore, must be analyzed considering both animal production and conservation.

Novel Ehrlichia and Hepatozoon genotypes in white-eared opossums (Didelphis albiventris) and associated ticks from Brazil.

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.

Trypanosomatid Richness Among Rats, Opossums, and Dogs in the Caatinga Biome, Northeast Brazil, a Former Endemic Area of Chagas Disease.

Parasites are important components of the immense n-dimensional trophic network that connects all living beings because they, among others, forge biodiversity and deeply influence ecological evolution and host behavior. In this sense, the influence of Trypanosomatidae remains unknown. The aim of this study was to determine trypanosomatid infection and richness in rats, opossums, and dogs in the semiarid Caatinga biome. We submitted DNA samples from trypanosomatids obtained through axenic cultures of the blood of these mammals to mini exon multiplex-PCR, Sanger, and next-generation sequencing targeting the 18S rDNA gene. Phylogenetic analyses were performed to identify genetic diversity in the Trypanosomatidae family. Shannon, Simpson, equability, and beta-diversity indices were calculated per location and per mammalian host. Dogs were surveyed for trypanosomatid infection through hemocultures and serological assays. The examined mammal species of this area of the Caatinga biome exhibited an enormous trypanosomatid species/genotypes richness. Ten denoised Operational Taxonomic Units (ZOTUs), including three species (Trypanosoma cruzi, Trypanosoma rangeli and Crithidia mellificae) and one Trypanosoma sp. five genotypes/lineages (T. cruzi DTU TcI, TcII, and TcIV; T. rangeli A and B) and four DTU TcI haplotypes (ZOTU1, ZOTU2, ZOTU5, and ZOTU10 merged), as well as 13 Amplicon Sequence Variants (ASVs), including five species (T. cruzi, T. rangeli, C. mellificae, Trypanosoma dionisii, and Trypanosoma lainsoni), five genotypes/lineages (same as the ZOTUs) and six DTU TcI haplotypes (ASV, ASV1, ASV2, ASV3, ASV5 and ASV13), were identified in single and mixed infections. We observed that trypanosomatids present a broad host spectrum given that species related to a single host are found in other mammals from different taxa. Concomitant infections between trypanosomatids and new host-parasite relationships have been reported, and this immense diversity in mammals raised questions, such as how this can influence the course of the infection in these animals and its transmissibility. Dogs demonstrated a high infection rate by T. cruzi as observed by positive serological results (92% in 2005 and 76% in 2007). The absence of positive parasitological tests confirmed their poor infectivity potential but their importance as sentinel hosts of T. cruzi transmission.

The outcomes of polyparasitism in stray cats from Brazilian Midwest assessed by epidemiological, hematological and pathological data.

We evaluated the epidemiological, hematological, and pathological data of Leishmania spp., Toxoplasma gondii, Platynosomum illiciens, feline immunodeficiency virus (FIV), and feline leukemia virus (FeLV) infections and the coinfections in stray cats of an endemic area for leishmaniasis. The diagnosis was performed by serological tests and necropsy. We described gross lesions and histopathological findings. We used immunohistochemistry and chromogenic in situ hybridization for L. infantum detection. We found infection in 27 out of 50 sampled cats, among them, 14 presented coinfections. A strong correlation between splenomegaly and lymphadenomegaly with FeLV, and an association between hepatic lesions and cachexia with parasitism due to P. illiciens were observed. Moreover, we found a significant increase in the monocyte count in the FeLV-infected and a decrease in the red blood cell count in the FIV-infected animals. Amastigote forms of Leishmania spp. and tissue changes were detected in lymphoid organs of an animal coinfected with P. illiciens, T. gondii, and FIV. Polyparasitism recorded in stray cats of the Brazilian Midwest should be considered in effective control strategies for public health diseases. Moreover, stray cats of Campo Grande may be a source of infection of FIV, FeLV and P. illiciens for populations of domiciled cats.

The influence of abiotic and biotic variables on the patent parasitemias of Trypanosoma spp. in Thrichomys fosteri (Rodentia: Echimyidae) in the southern Pantanal.

Parasitism is a dynamic ecological phenomenon that is constantly influenced by the environment and intrinsic factors of the host. We aimed to evaluate the influence of vegetation, environmental temperature, reproductive conditions, sex, and body condition (BC) on the detection of Trypanosoma spp. in the blood of Thrichomys fosteri in the Pantanal region, an enzootic area for trypanosomiasis. Whole blood was collected from the tip of the tail, and nPCR was performed for Trypanosoma spp. detection from the DNA extracted from the resultant blood clot. Statistical analyses were performed using generalized linear models. Our results showed that there is a greater probability of detection of Trypanosoma spp. in the bloodstream of animals with the highest BC values in periods with mild temperatures. Since T. fosteri is an abundant and common prey for carnivores, even in periods with low temperatures and consequent decrease in the reproduction and activities of the blood-sucking arthropod vectors, the maintenance of Trypanosoma spp. in the studied area would be guaranteed via predation (trophic network) of T. fosteri individuals with good BC and patent parasitemia. Furthermore, T. fosteri, which displays Trypanosoma spp. in the bloodstream, would be reproducing adequately because we found no influence between the reproductive condition and the detection of Trypanosoma spp. in T. fosteri. The caviomorph rodent T. fostei is an important species for the maintenance of Trypanosoma spp. in the Pantanal biome.

Leishmania infantum infecting the carnivore Nasua nasua from urban forest fragments in an endemic area of visceral leishmaniasis in Brazilian Midwest.

Introduction: The aim of the present study was to investigate the occurrence of Leishmania infantum in South American coatis inhabiting two forest fragments in Campo Grande, Mato Grosso do Sul, Midwest region of Brazil, an endemic area of human and canine visceral leishmaniasis (VL). Material and methods: A total of 110 South American coatis were sampled in the conservation unit "Parque Estadual do Prosa" (PEP) and in the residential area "Vila da Base Aérea" (VBA) from March 2018 to April 2019. As a longitudinal study that include up to six recaptures of the same individual, a total of 190 capture events were obtained. Blood, bone marrow and skin samples were obtained for parasitological (axenic culture), serological (Enzyme Linked Immunosorbent Assay - ELISA and Dual-path Platform immunoassay - DPP® CVL) and molecular diagnostic assays (targeting kDNA for Leishmania spp. and L. infantum; and HSP70 followed by sequence analysis). Results: Seropositivity for L. infantum was found in 33 individuals, six in PEP and 27 in VBA. Furthermore, L. infantum was detected by molecular analysis in 16 individuals, seven from PEP and nine from VBA. We also isolated L. infantum from bone marrow of one individual and detected a single positive skin sample in molecular assay from other individual, both from VBA. Discussion: An overall infection rate of 36.4% (40/110) was observed, significantly higher in the VBA (49.1%) than in the PEP (21.6%), probably because VBA presents: (i) a large number of resident dogs and chickens that would be attracting sandflies; (ii) a denser population of this wild mammal species; and (iii) physical barriers and a lack of functional connectivity in the surroundings, preventing these animals to disperse out. We conclude that South American coati populations living in urban forest fragments of Campo Grande are affected by the epidemiological scenario of VL, known to involve dogs, vectors and humans. We highlight the importance of investigate the parasitism by L. infantum in this and other potential L. infantum reservoirs that inhabit urbanized regions endemic to VL.

Molecular detection of piroplasmids in synanthropic rodents, marsupials, and associated ticks from Brazil, with phylogenetic inference of a putative novel Babesia sp. from white-eared opossum (Didelphis albiventris).

The order Piroplasmida encompasses tick-borne pathogens of veterinary and medical importance positioned in two main families: Babesiidae and Theileriidae. Even though previous studies carried out in Brazil recorded the occurrence of piroplasmid species circulating in small mammals, 18S RNA gene sequences were only partially sequenced, preventing the assessment of their phylogenetic positioning. The current study aimed to detect and characterize, using morphological, molecular, and bioinformatic approaches, piroplasmids from wild mammals and associated ticks sampled in Central-Western Brazil. Out of 67 Didelphis albiventris sampled, 22 (16.4%) were positive for piroplasmids by PCR. In contrast, none of the 48 small rodents and 14 capybaras (Hydrochoerus hydrochaeris) was PCR-positive. Four Amblyomma dubitatum ticks-one from Rattus rattus, one from H. hydrochaeris, and two from D. albiventris-out of 114 Amblyomma spp. DNA samples were positive for piroplasmids by PCR. The phylogenetic inference performed using the near-complete 18S rRNA gene positioned the putative novel piroplasmid species detected in D. albiventris and associated A. dubitatum ticks near to Babesia sensu lato clade (Western group-cluster III) and distant from the Australian marsupial-associated piroplasms. Phylogenetic inferences based on two additional molecular markers, namely hsp-70 and cox-1, supported the near-complete 18S rRNA gene phylogenetic inference. Finally, the partial 18S rRNA gene sequences detected in ticks from rodents (R. rattus and H. hydrochaeris) showed 97.2-99.4% identity with the Piroplasmida previously detected in a capybara from Brazil, raising evidence that a still uncharacterized piroplasmid species has been identified in the capybara, the largest rodent species from South America.

Relationships between vector-borne parasites and free-living mammals at the Brazilian Pantanal.

In nature, parasitic infections must be addressed as complex systems involving parasite-host relationships on a temporal and spatial scale. Since the parasites cover a great biological diversity, we can expect that wildlife are exposed simultaneously to different parasites. In this sense, the objective of this work was to determine the relationships between free-living mammals and their associated hemoparasites in the Brazilian Pantanal. We used the data published during 2017 and 2018 by de Sousa et al. regarding the detection of vector-borne pathogens (VBP), namely Anaplasma, Babesia, Bartonella, Cytauxzoon, Ehrlichia, Hepatozoon, Mycoplasma, and Theileria, in nine species of free-living mammals belonging to orders Carnivora, Rodentia, and Didelphimorphia. We assume as infected an individual positive on any of parasitological, molecular, and/or serological tests. We observed a strong association between the wild felid Leopardus pardalis with Cytauxzoon, the wild canid Cerdocyon thous with Hepatozoon, the small rodent Thrichomys fosteri with Bartonella, and the procyonid Nasua nasua with Mycoplasma and Theileria. Therefore, N. nasua, C. thous, T. fosteri, and the small rodent Oecomys mamorae can be considered key species for the maintenance of selected VBP in the Pantanal region, because they showed a high number of single and coinfections. Together, our results highlighted the importance of coinfection as a common phenomenon in nature.

Trypanosomatid species in Didelphis albiventris from urban forest fragments.

Urbanization results in loss of natural habitats and, consequently, reduction of richness and abundance of specialist to the detriment of generalist species. We hypothesized that a greater richness of trypanosomatid in Didelphis albiventris would be found in fragments of urban forests in Campo Grande, Mato Grosso do Sul, Brazil, that presented a larger richness of small mammals. We used parasitological, molecular, and serological methods to detect Trypanosoma spp. infection in D. albiventris (n = 43) from forest fragments. PCR was performed with primers specific for 18S rDNA, 24Sα rDNA, mini-chromosome satellites, and mini-exon genes. IFAT was used to detect anti-Trypanosoma cruzi IgG. All hemoculture was negative. We detected trypanosomatid DNA in blood of 35% of opossum. Two opossums were seropositive for T. cruzi. The trypanosomatid species number infecting D. albiventris was higher in the areas with greater abundance, rather than richness of small mammals. We found D. albiventris parasitized by T. cruzi in single and co-infections with Leishmania spp., recently described molecular operational taxonomic unit (MOTU) named DID, and Trypanosoma lainsoni. We concluded that (i) trypanosome richness may be determined by small mammal abundance, (ii) D. albiventris confirmed to be bio-accumulators of trypanosomatids, and (iii) T. lainsoni demonstrated a higher host range than described up to the present.

Low occurrence of Bartonella in synanthropic mammals and associated ectoparasites in peri-urban areas from Central-Western and Southern Brazil.

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.