Densidade parasitária e potencial ornitocórico de aves silvestres: resultados preliminares de um projeto de pesquisa e extensão realizado em área de reflorestamento no Campus IFRJ, Pinheiral, Rio de Janeiro, Brasil

This publication aims to disseminate to the scientific community and society in general, the project of the same title of this publication that is under development by the collaboration of the Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ) and the Universidade Federal Rural do Rio de aneiro (UFRRJ). Briefly, this project aims to quantify and identify parasites of the body surface (ectoparasites) and intestinal parasites (enteroparasites) of wild birds, which should be related to their biological and ecological conditions, and to evaluate the dispersal seed process performed by birds (ornithocoria), the viability and development of these seeds and thus establish the degree of importance of ornithocoria in reforestation at the IFRJ campus of Pinheiral, RJ. The project includes the integration of five doctoral professors, a laboratory technician doctor, as well as a postdoctoral student, four doctoral students, one master student, two undergraduate students and four technical high school students. To date, six expeditions have been made in the reforestation area of the Espaço Ecológico Educativo (EEcoE) of the IFRJ campus in Pinheiral, RJ, where a total of 196 wild birds have been captured, evaluated, ringed and recorded at the Centro Nacional de Pesquisa e Conservação de Aves Silvestres (CEMAVE), of which 29 were recaptured for revaluations. Four articles were published on the themes of parasitology and ornithochory in wild birds. Two practical short courses in the area of study were taught in September 2018 and February 2019 to IFRJ students, but open to the scientific community and society at large. Currently the project is not expected to end, since there are plenty of unpublished and relevant results in both the area of parasitology and ornithochory of wild birds.A presente publicação tem por objetivo divulgar, à comunidade científica e à sociedade em geral, o projeto de mesmo título desta publicação que está em desenvolvimento em colaboração dos Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ) e Universidade Federal Rural do Rio de Janeiro (UFRRJ). Resumidamente, este projeto objetiva quantificar e identificar parasitas da superfície corporal (ectoparasitas) e parasitas intestinais (enteroparasitas) de aves silvestres, os quais devem estar relacionados às suas condições biológicas e ecológicas, além de avaliar o processo de dispersão de sementes desempenhado pelas aves (ornitocoria), a viabilidade e desenvolvimento destas sementes e, desta forma, estabelecer o grau de importância da ornitocoria no reflorestamento no campus IFRJ de Pinheiral, RJ. O projeto conta com a integração de cinco professores doutores, uma doutora técnica de laboratório, além de um pós-doutorando, quatro doutorandos, um mestrando, dois estudantes de iniciação científica e quatro estudantes de ensino médio-técnico. Até o presente momento, foram realizadas seis expedições na área em reflorestamento do Espaço Ecológico Educativo (EEcoE) do campus IFRJ em Pinheiral, RJ, onde o total de 196 aves silvestres foram capturadas, avaliadas, anilhadas e registradas no Centro Nacional de Pesquisa e Conservação de Aves Silvestres (CEMAVE), das quais 29 foram recapturadas para reavaliações. Quatro artigos foram publicados nos temas de parasitologia e ornitocoria em aves silvestres. Dois minicursos práticos, na área de estudo, foram ministrados em setembro de 2018 e fevereiro de 2019 para alunos do IFRJ, porém aberto para a comunidade científica e sociedade em geral. Atualmente o projeto não tem previsão para término, desde que há abundância de resultados inéditos e relevantes tanto na área de parasitologia, quanto ornitocoria em aves silvestres

Isospora sepetibensis Berto, Flausino, Luz, Ferreira, Lopes 2008

Isospora sepetibensis Berto, Flausino, Luz, Ferreira, Lopes, 2008 (Fig. 1) Description of sporulated oocyst: Oocyst shape (n = 18) sub-spherical to elongate ovoidal; 22–29 × 19–22 (25.9 × 20.7) length/width (L/W) ratio 1.1–1.4 (1.26). Wall bi-layered, 1.1–2.0 (1.3) thick, outer layer smooth, c. 2/3 of total thickness. Micropyle and oocyst residuum absent, but one or two polar granules are present. Description of sporocyst and sporozoites: Sporocysts (n = 18), ellipsoidal, 15–18 × 9–11 (16.8 × 10.3); L/W ratio 1.5–1.8 (1.65). Stieda body present, knob-like, 1.1–1.3 (1.2) high, 2.0–2.2 (2.1) wide; sub-Stieda body present, large to trapezoidal or, occasionally, irregular, 1.2–2.3 (1.7) high, 2.7–3.5 (3.1) wide; para-Stieda body absent. Sporocyst residuum present, consisting of numerous small granules dispersed between the sporozoites or as a distinctly sub-spherical body that appear to be membrane-bounded, 5.5–6.7 (6.1). Sporozoite vermiform with one posterior refractile body, centrally located nucleus and striations. Host: Trichothraupis melanops (Vieillot, 1818) (Passeriformes: Thraupidae: Tachyphoninae). Locality: Itatiaia National Park (22°26’17.00”S, 44°37’33.00”W), Southeastern Brazil. Specimens: Photomicrographs and oocysts in 2.5% K 2 Cr 2 O 7 solution (Williams et al. 2010) are deposited at the Museu de Zoologia at the Universidade Federal Ru- ral do Rio de Janeiro, Brazil, under accession number MZURPTZ2019014. Photomicrographs are also de- posited and available (http://r1.ufrrj.br/labicoc/colecao. html) in the Parasitology Collection of the Laboratório de Biologia de Coccídios, at UFRRJ, under repository number 92/2019. Photographs of the host specimens are deposited in the same collection. Site in host: Unknown. Prevalence: 67% (15 out of 10 birds infected). Representative DNA sequence: Representative cox1 sequence is deposited in the GenBank database under the accession number MK682606. Phylogenetic analysis: The amplification of the DNA from the fifteen oocysts of I. sepetibensis recovered from T. melanops showed a clear band of c. 250 bp. The phylogenetic analysis was constructed with 33 sequences of Isospora spp. closest to I. sepetibensis available on GenBank (Fig. 2). Eimeria tenella (Railliet, Lucet, 1891) was used as the outgroup. Isospora sepetibensis sat in a large group with the highest similarity of 98.1% with an Isospora sp. reported as pseudoparasite of bank voles Myodes glareolus (Schreber, 1780) in Czech Republic (Trefancová et al. 2019). It was close to the clade with Isospora isolates from the spectacled warbler Sylvia conspicillata Temminck, 1820 in Macaronesia (Illera et al. 2015); and also was close to the clade with Isospora spp. recently sequenced from Neotropical passerines (Silva-Carvalho et al. 2018a, 2018b; Rodrigues et al. 2019). Subsequently, a subset with only 215 bp long cox1 gene sequences was constructed (Fig. 3). In this analysis, Isospora sepetibensis sat in the same clade of Isospora spp. from Neotropical passerines, with the highest similarity of 98.5% with Isospora lopesi Silva-Carvalho & Berto, 2018 (Silva-Carvalho et al. 2018a).Published as part of Genovez-Oliveira, Jhon Lennon, Cardozo, Sergian V., De Oliveira, Águida A., De Lima, Viviane M., Ferreira, Ildemar & Berto, Bruno P., 2019, Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America, pp. 17-23 in Acta Protozoologica 58 (1) on pages 18-19, DOI: 10.4467/16890027AP.19.007.10838, http://zenodo.org/record/835693

Fig. 2. Maximum likelihood tree estimated from the cox1 in Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America

Fig. 2. Maximum likelihood tree estimated from the cox1 sequences. Numbers at nodes represent bootstrap support (1,000 replicates; only values> 50% shown) for Neighbor-Joining and Maximum Likelihood, respectively. The scale-bar represents the number of nucleotide substitutions per site.Published as part of Genovez-Oliveira, Jhon Lennon, Cardozo, Sergian V., De Oliveira, Águida A., De Lima, Viviane M., Ferreira, Ildemar & Berto, Bruno P., 2019, Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America, pp. 17-23 in Acta Protozoologica 58 (1) on page 20, DOI: 10.4467/16890027AP.19.007.10838, http://zenodo.org/record/835693

Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America

Genovez-Oliveira, Jhon Lennon, Cardozo, Sergian V., De Oliveira, Águida A., De Lima, Viviane M., Ferreira, Ildemar, Berto, Bruno P. (2019): Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America. Acta Protozoologica 58 (1): 17-23, DOI: 10.4467/16890027AP.19.007.10838, URL: http://dx.doi.org/10.4467/16890027ap.19.007.1083

Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America

Isospora sepetibensis Berto, Flausino, Luz, Ferreira and Lopes, 2008 is a protozoan coccidian parasite (Chromista: Miozoa: Coccidiomorphea: Coccidia) that was originally described from Brazilian tanagers Ramphocelus bresilius (Linnaeus, 1766) in the Marambaia Island in the Coast of the State of Rio de Janeiro. In the current  work, this species was identified from black-goggled tanagers Trichothraupis melanops (Vieillot, 1818) in the Itatiaia National Park, which is a protected area with a high degree of vulnerability in the interior of the State of Rio de Janeiro, distant in more than 100 km of the type-locality. Its oocysts are sub-spherical to elongate  ovoidal, 25.9 × 20.7 μm with smooth, bi-layered wall, ~ 1.3 μm and length/width ratio of 1.1–1.4 (1.26). Micropyle and oocyst residuum absent, but one or two polar granules are present. Sporocysts are ellipsoidal, 16.8 × 10.3 μm, with both Stieda and sub-Stieda bodies. Sporocyst residuum present  and sporozoites with refractile body and nucleus. Molecular analysis was conducted at the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. This new isolate exhibited similarity greater than 98% with Isospora spp. isolates from other Neotropical passerines and with an Isospora sp. pseudoparasite of voles of Eurasia. This is the first coccidian  parasite from a New World tanager to have a molecular identification of the cox1 gene

Fig. 3. Maximum likelihood tree estimated from the 215 in Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America

Fig. 3. Maximum likelihood tree estimated from the 215 bp long cox1 sequences. Numbers at nodes represent bootstrap support (1,000 replicates; only values> 50% shown) for Neighbor-Joining and Maximum Likelihood, respectively. The scale-bar represents the number of nucleotide substitutions per site.Published as part of Genovez-Oliveira, Jhon Lennon, Cardozo, Sergian V., De Oliveira, Águida A., De Lima, Viviane M., Ferreira, Ildemar & Berto, Bruno P., 2019, Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America, pp. 17-23 in Acta Protozoologica 58 (1) on page 21, DOI: 10.4467/16890027AP.19.007.10838, http://zenodo.org/record/835693

Fig. 1 in Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America

Fig. 1. Photomicrographs of sporulated oocysts of Isospora sepetibensis, a coccidium species recovered from the black-goggled tanager Trichothraupis melanops. Note the inner (il) and outer (ol) layer of the oocyst wall, nucleus (n), polar granule (pg), Stieda body (sb), sub- Stieda body (ssb), sporocyst residuum (sr), striations (str) and the refractile body (rb). Sheather's sugar solution. Scale-bar: 10 µm.Published as part of Genovez-Oliveira, Jhon Lennon, Cardozo, Sergian V., De Oliveira, Águida A., De Lima, Viviane M., Ferreira, Ildemar & Berto, Bruno P., 2019, Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America, pp. 17-23 in Acta Protozoologica 58 (1) on page 19, DOI: 10.4467/16890027AP.19.007.10838, http://zenodo.org/record/835693