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French Guiana, located in the Guiana Shield, is a natural reservoir for many zoonotic pathogens that are of considerable medical or veterinary importance. Until now, there has been limited data available on the description of parasites circulating in this area, especially on protozoan belonging to the phylum Apicomplexa; conversely, the neighbouring countries describe a high parasitic prevalence in animals and humans. Epidemiological surveillance is necessary, as new potentially virulent strains may emerge from these forest ecosystems, such as Amazonian toxoplasmosis. However, there is no standard tool for detecting protozoa in wildlife. In this study, we developed Meat-Borne-Parasite, a high-throughput meta-barcoding workflow for detecting Apicomplexa based on the Oxford Nanopore Technologies sequencing platform using the 18S gene of 14 Apicomplexa positive samples collected in French Guiana. Sequencing reads were then analysed with MetONTIIME pipeline. Thanks to a scoring rule, we were able to classify 10 samples out of 14 as Apicomplexa positive and reveal the presence of co-carriages. The same samples were also sequenced with the Illumina platform for validation purposes. For samples identified as Apicomplexa positive by both platforms, a strong positive correlation at up to the genus level was reported. Overall, the presented workflow represents a reliable method for Apicomplexa detection, which may pave the way for more comprehensive biomonitoring of zoonotic pathogens.

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The objective of this study was to evaluate the occurrence of Clostridium perfringens in stool samples and swabs collected from wild mammals in the Amazon biome. Sixty-five faecal and swab samples were collected in situ and ex situ from 16 species and three genera of wild mammals, some of which were in good health and some of which had diarrhoea. After pre-enrichment, the samples were plated on selective agar for C. perfringens. Characteristic colonies were subjected to multiplex PCR for the detection of genes encoding the main C. perfringens toxins (alpha, beta, epsilon, and iota toxin and enterotoxin). Among the 65 samples, 40 (61.5%) were positive for the gene encoding the alpha toxin and were classified as type A, 36 of which were asymptomatic animals and four were diarrheal. No other toxinotypes were found. The findings of this study suggest that C. perfringens type A is commonly found in mammal species of the Amazon biome. This seems to be the first study to identify C. perfringens type A in species such as B. variegatus (common ground sloth), C. didactylus (two-toed sloth), P. flavus (Jupará), T. tetradactyla (anteater), S. collinsi (squirrel monkey), S. niger (black marmoset), and S. apella (Guyana capuchin) and in the genus Didelphis sp. (opossum).

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Trypanosoma cruzi hosts can serve as a source of infection for animals, vectors, and humans, contributing to the establishment of Chagas disease (CD) in a given area. Traditionally, the Department of Córdoba has not been considered a transmission area for CD; however, the report of several acute cases of Chagas disease highlights the importance of studying the dynamics of disease transmission in this region. This study aimed to detect T. cruzi in domestic and wild mammals in the department of Córdoba. In 2017, a cross-sectional descriptive study was conducted in six villages in two municipalities in the department of Córdoba. Blood samples from dogs living in the zones were collected in EDTA vacutainer tubes for domestic mammals. Wild mammals were collected using Sherman and Tomahawk traps and mist nets in crops and peridomiciles. T. cruzi DNA was detected using the kinetoplast DNA (kDNA) variable region and the tandem repeat satellite region of T. cruzi as molecular targets. We sampled 168 dogs and 146 wild mammals. The detected prevalence of T. cruzi was 6.37%; the TcI lineage was found in D. marsupialis, H. anomalus, and one canine. A specimen of D. marsupialis with TcI and TcII lineages was also identified. T. cruzi DNA was detected in domestic and wild animals in the study area, indicating the circulation of the parasite in peridomestic environments. D. marsupialis may represent an important host in maintaining this region's wild and domestic cycle.

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BACKGROUND: The study of the ecology of Trypanosoma cruzi is challenging due to its extreme adaptive plasticity, resulting in the parasitism of hundreds of mammal species and dozens of triatomine species. The genetic analysis of blood meal sources (BMS) from the triatomine vector is an accurate and practical approach for gathering information on which wild mammal species participate in a local transmission network. South American coatis, Nasua nasua, act as important reservoir host species of T. cruzi in the Pantanal biome because of their high rate of infection and elevated parasitemia, with the main discrete typing unit (DTU) lineages (TcI and TcII). Moreover, the carnivore coati is the only mammal species to build high arboreal nests for breeding and resting that can be shared by various vertebrate and invertebrate species. Herein, we applied the sensitive and specific methodology of DNA barcoding and molecular cloning to study triatomines found in a coati nest to access the diversity of mammal species that explore this structure, and therefore, may be involved in the parasite transmission network. METHODS: Twenty-three Triatoma sordida were collected in one coati's nest in the subregion of Nhecolândia, Pantanal. The DNA isolated from the gut of insects was subjected to BMS detection by PCR using universal primers that flank variable regions of the cytochrome b (cytb) and 12S rDNA mitochondrial genes from vertebrates. The Trypanosoma spp. diagnosis and DTU genotyping were based on an 18S rDNA molecular marker and also using new cytb gene primers designed in this study. Phylogenetic analyses and chord diagrams were constructed to visualize BMS haplotypes, DTU lineages detected on vectors, and their interconnections. RESULTS: Twenty of 23 triatomines analyzed were PCR-positive (86.95%) showing lineages T. cruzi DTU TcI (n = 2), TcII (n = 6), and a predominance of TcI/TcII (n = 12) mixed infection. Intra-DTU diversity was observed mainly from different TcI haplotypes. Genetic analyses revealed that the southern anteater, Tamandua tetradactyla, was the unique species detected as the BMS of triatomines collected from the coati's nest. At least three different individuals of T. tetradactyla served as BMS of 21/23 bugs studied, as indicated by the cytb and 12S rDNA haplotypes identified. CONCLUSIONS: The identification of multiple BMS, and importantly, different individuals of the same species, was achieved by the methodology applied. The study demonstrated that the southern anteaters can occupy the South American coati's nest, serving as the BMS of T. sordida specimens. Since anteaters have an individualist nonsocial behavior, the three individuals detected as BMS stayed at the coati's nest at different times, which added a temporal character to BMS detection. The TcI and TcII infection, and significantly, a predominance of TcI/TcII mixed infection profile with different TcI and TcII haplotypes was observed, due to the discriminatory capacity of the methodology applied. Tamandua tetradactyla, a host which has been little studied, may have an important role in the T. cruzi transmission in that Pantanal subregion. The data from the present study indicate the sharing of coatis' nests by other mammal species, expanding the possibilities for T. cruzi transmission in the canopy strata. We propose that coatis' nests can act as the true hubs of the T. cruzi transmission web in Pantanal, instead of the coatis themselves, as previously suggested.

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PURPOSE: Leishmaniasis are infectious and zoonotic diseases and present in cutaneous and visceral forms. Cutaneous leishmaniasis is endemic and widely distributed throughout the state of Espírito Santo, Brazil. Several cases of cutaneous leishmaniasis in humans and dogs associated with Leishmania (Viannia) braziliensis have been reported in the state. This study aimed to investigate the occurrence of natural infection by Leishmania(Viannia) braziliensis in wild mammals found dead (by trampling or natural death) in the Sooretama Biological Reserve, Espírito Santo State. METHODS: From January 2018 to December 2019, 60 animals were collected. Of these, 47 animals from 12 different species were analyzed. The results were demonstrated using descriptive analysis of the observations to calculate the absolute and relative frequencies of the data. RESULTS: In the PCR, using specific primers for the genus Leishmania (D1, D2, and D3) and the species Leishmania (Viannia) braziliensis (ISVB/ISVC), 4 positive animals (8.5%) were detected: 1 Cuniculus paca (paca) (25%) and 3 Callithrix geoffroyi (white-faced marmoset) (25%). In the histopathological analysis, the parasitic amastigote form was not observed. CONCLUSIONS: The natural infection, detected by PCR, by Leishmania (Viannia) braziliensis in Cuniculus paca (paca) and Callithrix geoffroyi (white-faced marmoset) constitutes the first report of infection of this rodent and primate species in the literature. Despite the confirmation of Leishmania (Viannia) braziliensis infection in rodents and primates, the role of these species in the transmission of this zoonosis still needs further observational studies to identify their seasonal variation, transmissibility, infection stability, and the effects of a given parasite on the population and/or individual.

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The Iguaçu National Park (INP) is the largest remnant of Atlantic Forest in southern Brazil, representing an ecological continuum with Argentina. The INP harbours a diverse fauna, with ring-tailed coatis (Nasua nasua Linnaeus, 1976, Carnivora: Procyonidae) in close contact with tourists either begging and/or snatching food from visitors. A potentially novel haemotropic Mycoplasma sp. has been previously detected in the ring-tailed coatis from central-western and southern Brazil. Therefore, the aims of this study were to investigate the occurrence of haemotropic Mycoplasma sp. and tick-borne pathogens in wild ring-tailed coatis from the INP, Foz do Iguaçu municipality, Paraná State, southern Brazil. Blood samples were collected from 18 wild ring-tailed coatis and evaluated by conventional PCR (cPCR) assays for haemotropic Mycoplasma spp. (16S and 23S rRNA), Theileria/Babesia spp. (18S rRNA) and Ehrlichia/Anaplasma spp. (16S rRNA, sodB, dsb and groEL). Eight out of 18 (44.44%; 95% CI: 24.56%-66.28%) animals were positive for haemotropic Mycoplasma spp. All ring-tailed coatis tested negative for Theileria/Babesia spp. and only one out of 18 (5.56%; 95% CI: 0.99%-25.76%) animals tested positive for Ehrlichia/Anaplasma spp. by the 16S rRNA cPCR. Unfortunately, multiple attempts to sequence the 16S rRNA gene of the Ehrlichia/Anaplasma-positive sample have failed. Phylogenetic and network analysis of the hemoplasma 16S and 23S rRNA gene fragments confirmed that animals were infected by a potentially novel haemotropic Mycoplasma sp. previously reported in ring-tailed coatis from Brazil. The name 'Candidatus Mycoplasma haematonasua' is proposed for this novel organism.

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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.

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Ovarian response of collared peccaries (Pecari tajacu), after hormonal stimulation with gonadotropin association (eCG/hCG), was accessed by both gene expression and follicular development. Thus, collared peccaries (n = 8) were treated with the dose used for sows (swine dose, SWD) or with dose adjusted for peccary's weight (allometric dose, ALD). The gene expression of receptors was evaluated for both gonadotropins (FSHR and LHCGR) and growth factors (proteins codified by TGFßR-1, BMPR1-A and BMPR2 genes) in antral follicles, cortex and corpora haemorrhagica (CH). Five days after gonadotropin injection, all females presented CH. The ovulation rate was similar (p > .05) between SWD (4.00 ± 1.17) and ALD (2.50 ± 0.43) group. The total number of follicles per animal and amounts of small (<3 mm), medium (3-5 mm) and large (>5 mm) follicles was similar among groups. However, SWD produced large follicles heavier than ALD group, as accessed by weight of follicular wall biopsies. Ovarian follicles expressed both gonadotropin and growth factor receptors at levels which are independent from gonadotropin dose. In conclusion, the two gonadotropin doses (SWD and ALD) can be used for ovarian stimulation of collared peccary. Additionally, FSH and growth factors (TGFßR-1, BMPR1-A and BMPR2) receptors are more expressed in the early follicle development, while LH receptor seems to be more important in the final of follicular growth.

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ABSTRACT: Arboviruses are viruses that maintain their life cycle in the wild and are transmitted to vertebrate hosts by hematophagous diptera. They are zoonotic and can establish an enzootic cycle in the urban areas; in humans, the infection can manifest from being encephalitogenic to hemorrhagic. This study aimed to report the occurrence of arboviruses in mammals of the order Didelphimorphia and Rodentia, captured from the Amazon. Serum samples were subjected to hemagglutination inhibition test using a viral panel of 19 species of arboviruses that are known to occur in the Amazon. Altogether, 14 wild mammals, 12 of Philander opossum, 1 of Didelphis marsupialis, and 1 of Nectomys rattus were captured. Eight of these were reported to be seropositive for arboviruses (57.14%) with monotypic seroprevalence for the Eastern Equine Encephalitis Virus (n=1), the Ilheus Virus (n=2), and the Catu virus (n=4); 4 heterotypic responses were observed for Flavivirus and Orthobunyavirus. In conclusion, several arbovirus species are in active circulation and maintenance, exhibiting enzootic characteristics in the wild mammals of the Amazon region; these animals prove to be potential hosts in the transmission of diseases to humans.

RESUMO: Os arbovírus são vírus que mantêm o seu ciclo de vida em ambiente silvestre. Eles são transmitidos aos hospedeiros vertebrados por dípteros hematófagos, tem caráter zoonótico podendo estabelecer um ciclo enzoótico no meio urbano, sendo que em humanos a infecção pode ter caráter encefalomiogênico a hemorrágico. Este estudo teve como objetivo relatar a ocorrência de arbovírus em mamíferos da ordem Didelphimorphia e Rodentia capturados na Amazônia. Os soros dos animais foram submetidos a testes de Inibição da Hemaglutinação utilizando um painel viral com as 19 principais espécies de arbovírus que ocorrem na Amazônia. Foram capturados 14 espécimes de mamíferos silvestres, 12 Philander opossum, 1 Didelphis marsupialis e 1 Nectomys rattus. A soropositividade para arbovírus foi observada em 57,14% (8/14) dos espécimes estudados, com soroprevalência monotípica para o vírus da Encefalite Equina do Leste (n = 1), o vírus Ilheus (n = 2) e o vírus Catu (n = 4) e quatro reações heterotípicos para Flavivírus e Orthobyavírus. Conclui-se que há manutenção e circulação de espécies de arbovírus com características enzoóticas em mamíferos silvestres da região amazônica, podendo ser hospedeiros em potenciais na transmissão da doença para humanos.

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Distintas estratégias de conservação têm sido adotadas visando a manutenção e recuperação da biodiversidade, especialmente de espécies que se encontram em diferentes níveis de ameaça à extinção. Neste grupo de espécies, encontram-se os grandes felídeos, os quais em virtude das ações antrópicas, como a destruição e a fragmentação de habitat, necessitam de esforços voltados para a conservação de seu material genético. Uma estratégia empregada para essa finalidade consiste nos bancos de recursos somáticos, os quais são definidos como a criopreservação de tecidos e células somáticas oriundas de diferentes populações. Essas amostras biológicas, quando adequadamente conservadas, são elementoschave para a multiplicação das espécies por meio de seu emprego na clonagem por transferência nuclear e indução de células à pluripotência. Assim, o objetivo desta revisão é abordar os aspectos relevantes envolvidos na formação de bancos de recursos somáticos em grandes felídeos, destacando os estudos desenvolvidos pelo Laboratório de Biotecnologia Animal (LBA), da Universidade Federal Rural do Semi-Árido (UFERSA) em onças-pintadas e onças-pardas.

Different conservation strategies have been adopted to maintain and recover biodiversity, especially for species that are at different levels of threat to extinction. In this group of species are the large felids, which, due to anthropic actions, such as habitat destruction and fragmentation, require efforts aimed at the conservation of their genetic material. A strategy employed for this purpose consists of somatic resource banks, which are defined as the cryopreservation of tissues and somatic cells from different populations. These biological samples, when properly preserved, are key elements for the multiplication of species through their use in cloning by nuclear transfer and induction of cells to pluripotency. Thus, the aim of this review is to address the relevant aspects involved in the formation of somatic resource banks in large felids, highlighting the studies carried out by the Laboratory of Animal Biotechnology (LBA) of the Federal Rural University of the Semi-Arid (UFERSA) in jaguars and pumas.

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