Redescription and molecular identification of Isospora feroxis Berto, Luz, Flausino, Ferreira & Lopes, 2009 (Eimeriidae) from tyrant-flycatchers (Tyrannoidea) in South America.

In the present study Isospora feroxis Berto, Luz, Flausino, Ferreira & Lopes, 2009 is redescribed from the photosyntypes and from new samples from a short-crested flycatcher Myiarchus ferox (Gmelin), which is the type-host in the type-locality, the Marambaia Island in Southeastern Brazil. In addition, the yellow-olive flycatcher Tolmomyias sulphurescens Spix is recorded as a new host for this species, in a new locality, the Itatiaia National Park, in the interior of Southeastern Brazil, providing a preliminary genotypic characterization via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene. Micropyle and rough oöcyst wall are added to the description of I. feroxis, in addition to other details. This is the sixth species identified from suboscine birds (Tyranni) to have a COI gene sequence deposited in GenBank and, although it is not yet possible to make conclusions on the phylogeny of Isospora spp. from Passeriformes by the COI gene, the molecular analysis confirmed the differences between coccidian species from tyrant-flycatchers.

Eimeria ferreirai n. sp. (Chromista: Miozoa: Eimeriidae) from doves Leptotila spp. (Columbiformes: Columbidae) from Brazil.

The doves and pigeons constitute a taxonomic group (Columbiformes: Columbidae) of granivorous and frugivorous birds with a worldwide distribution. The current work aims to describe morphologically and molecularly a new protozoan from white-tipped doves Leptotila verreauxi Bonaparte, 1855 and grey-fronted doves Leptotila rufaxilla (Richard Bernard, 1792) in Southeastern Brazil. Eimeria ferreirai n. sp. has oocysts that are sub-spherical to ellipsoidal, 21.4 × 18.8 µm, with smooth, bilayered wall, ~1.6 µm thick. Micropyle present. Oocyst residuum absent, but one to two polar granules are present. Sporocysts are elongate ovoidal to boomerang-shaped, 13.4 × 6.9 µm. Stieda body triangular to lozengal. Sporocyst residuum is composed of granules of different sizes. Sporozoites are vermiform with refractile body and nucleus. Sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene and the subsequent phylogenetic molecular comparisons supported the description of the new species, since the maximum similarity was 90-95% with eimeriid species of Columbiformes, Anseriformes, Galliformes and Passeriformes. Thus, this is the first coccidian species reported from Leptotila spp. and the twentieth description of an eimerian from Columbiformes in the World.

Isospora oliveirai n. sp. (Chromista: Miozoa: Eimeriidae) from the Greenish Schiffornis Schiffornis virescens (Lafresnaye, 1838) (Passeriformes: Tyranni: Tityridae) in South America.

BACKGROUND: Coccidia are obligatory intracellular parasites with at least one intestinal phase in their life cycles, being Isospora Schneider, 1881 the main coccidian genus related to the order Passeriformes. However, there is no record of isosporans from the passerine family Tityridae, which is the family of the greenish schiffornis Schiffornis virescens (Lafresnaye, 1838). PURPOSE: This study aimed to examine the faeces from a greenish schiffornis S. virescens captured in the Itatiaia National Park, State of Rio de Janeiro, Southeastern Brazil, to determine what coccidian parasites were present. METHODS: Only one specimen of Schiffornis virescens was captured with mist nets. Coccidian oocysts were recovered from the fecal samples by flotation in Sheather's saturated solution. Morphological observations, line drawings, photomicrographs and measurements were made in optical microscopy and digitally edited. The molecular analysis included the study of the sequence of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene, with phylogenetic reconstructions based on the Neighbor-Joining and Maximum Likelihood analysis. RESULTS: An Isospora sp. considered as new to science is described and identified from Schiffornis virescens (Lafresnaye, 1838). Isospora oliveirai n. sp. has oocysts that are subspheroidal, 26.0 × 24.8 µm, with rough, bilayered wall, c.2.5 µm thick. Micropyle and oocyst residuum absent, but one to six polar granules are present. Sporocysts lemon-shaped, 18.1 × 10.9 µm. The Stieda body is knob-like to half-moon-shaped and sub-Stieda is rounded. Sporocyst residuum is present, composed of scattered spherules of different sizes. Sporozoites are vermiform, with refractile bodies and nucleus. Molecular analysis at the COI gene exhibited similarity of 97% with Isospora serinuse Yang, Brice, Elliot et Ryan, 2015 from island canaries Serinus canaria (Linnaeus, 1758), and Isospora spp. from great tits Parus major (Linnaeus, 1758) and European robins Erithacus rubecula (Linnaeus, 1758). CONCLUSION: Based on the morphological and molecular features, I. oliveirai is considered as new to science and the first coccidian species recorded from Tityridae.

Morphological and molecular identification of Isospora massardi Lopes, Berto, Luz, Galvão, Ferreira & Lopes, 2014 (Chromista: Miozoa: Eimeriidae) from thrushes Turdus spp. (Passeriformes: Turdidae) in South America.

In the current work, Isospora massardi Lopes, Berto, Luz, Galvão, Ferreira & Lopes, [10] is identified from white-necked thrushes Turdus albicollis Vieillot, 1818, rufous-bellied thrushes Turdus rufiventris Vieillot, 1818 and from a new host, the yellow-legged thrush Turdus flavipes (Vieillot, 1818) in a new locality, the Itatiaia National Park, in Southeastern Brazil, providing a preliminary genotypic characterization via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) and 18S small subunit ribosomal RNA genes. The oocysts and sporocysts of I. massardi of the current study are morphologically similar to the original description and are uniform in the proportionality of width on length, but exhibited different patterns of size associated with each host species. Furthermore, a genotypic difference of 3% was found in the COI sequences from T. flavipes and T. albicollis. Therefore, an ecological discussion is introduced aimed at associating these morphometric and genotypic differences with an ongoing speciation process.

Coccidia of Columbiformes: a taxonomic review of its Eimeriidae species and Eimeria columbinae n. sp. from Columbina talpacoti (Temminck, 1809) from Brazil.

Coccidia (Chromista: Miozoa: Eimeriidae) of columbiform birds (Aves: Columbiformes) have been described since the end of the nineteenth century; however, some of these descriptions were poorly detailed or inconclusive. In this sense, the current work makes a detailed taxonomic revision reconsidering and organizing 18 Eimeria spp. and two Isospora spp. previously described or reported of Columbiformes. Along with this, a new species of Eimeria is morphologically and molecularly identified by the mitochondrial cytochrome c oxidase subunit 1 (COI) gene and by the 18S small subunit ribosomal RNA (18S) gene from the ruddy ground-dove Columbina talpacoti (Temminck, 1809) in the Médio Paraíba region of the State of Rio de Janeiro, southeastern Brazil. Eimeria columbinae n. sp. has subspheroidal oocysts, 14.7 × 13.2 µm, with smooth, bi-layered wall, ~ 1.1 µm and length/width ratio of 1.1. Micropyle and oocyst residuum are present, but polar granule is absent. Sporocysts are ellipsoidal to slightly asymmetrical, 9.0 × 5.1 µm, with both Stieda and sub-Stieda bodies. Sporocyst residuum present and sporozoites with refractile body and nucleus. This is the 19th description of an eimerian from Columbiformes in the World, and the second to have a molecular identification of the COI and 18S genes.

Isospora borbai n. sp. (Chromista: Apicomplexa: Eimeriidae) from gnateaters Conopophaga spp. (Passeriformes: Tyranni: Conopophagidae) in South America.

BACKGROUND: The gnateaters Conopophaga spp. are insectivorous passerines commonly observed in high and humid forests, where they remain lodged in thin branches and, sometimes, they fly to the ground to catch insects. The insectivorous feeding habit is related to low prevalence and density of coccidians in passerines; however, several coccidian species are recorded for families of insectivorous passerines. PURPOSE: This study aimed to examine the feces from gnateaters Conopophaga spp. captured in the municipality of Barra Mansa and in the Itatiaia National Park, State of Rio de Janeiro, Southeastern Brazil, to determine what coccidian parasites were present. METHODS: Nine gnateaters were captured with mist nets. Coccidian oocysts were recovered from the fecal samples by flotation in Sheather's saturated solution. Morphological observations, line drawings, photomicrographs and measurements were made in optical microscopy and digitally edited. The molecular analysis included the study of the sequence of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene, with phylogenetic reconstructions based on the neighbor-joining and maximum likelihood analysis. RESULTS: Four Conopophaga spp. were positive for oocysts. An Isospora sp. considered as new to science is described and identified from Conopophaga melanops (Vieillot, 1818) and Conopophaga lineata (Wied, 1831). Isospora borbai n. sp. has oocysts that are subspheroidal, 17-22 × 15-22 (20.2 × 19.1) µm, with rough, bilayered wall, c.1.7 µm thick. Micropyle present, but without micropyle cap. Oocyst residuum absent, but one or two polar granules are present. Sporocysts are ellipsoidal, 12-15 × 8-11 (14.1 × 9.1) µm. The Stieda body is knob-like to half-moon-shaped and sub-Stieda body is rounded. Sporocyst residuum is present, composed of scattered spherules of different sizes. Sporozoites are vermiform with refractile body and nucleus. Molecular analysis at the cox1 gene exhibited similarity greater than 99% with Isospora spp. isolates from other Neotropical passerine birds. CONCLUSION: Based on the morphological and molecular features, I. borbai is considered as new to science and the first coccidian species recorded from Conopophagidae.