Morphological and Molecular Identification of Isospora sepetibensis (Chromista: Miozoa: Eimeriidae) from a New Host, Trichothraupis melanops (Passeriformes: Thraupidae: Tachyphoninae) in South America Author Genovez-Oliveira, Jhon Lennon Author Cardozo, Sergian V. Author De Oliveira, Águida A. Author De Lima, Viviane M. Author Ferreira, Ildemar Author Berto, Bruno P. text Acta Protozoologica 2019 58 1 17 23 http://dx.doi.org/10.4467/16890027ap.19.007.10838 journal article 10.4467/16890027AP.19.007.10838 1689-0027 8356932 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. 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. 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 ).