Isospora similisi recovered from a new host, Saltator aurantiirostris, with supplementary molecular data and notes on its taxonomy and distribution in the Neotropical region.

This article reports on a golden-billed saltator Saltator aurantiirostris Vieillot, 1817, kept in captivity outside its natural distribution area, in the proximities of the Itatiaia National Park, as a new host for Isospora similisi Coelho, Berto, Neves, Oliveira, Flausino & Lopes, 2013. Additionally, a supplementary molecular identification is provided through the sequencing of three non-overlapping loci of mitochondrial DNA and one locus of the 18S small subunit ribosomal RNA (18S) gene. All the taxonomic features of the I. similisi oocysts shed by S. aurantiirostris were equivalent to those originally described from Saltator similis d'Orbigny & Lafresnaye, 1837. The new sequenced loci were identical, or showed 99.9% similarity, to the samples of I. similisi from S. similis and S. aurantiirostris, confirming the same species from both hosts. Lastly, I. similisi is estimated as a junior synonym of Isospora formarum McQuistion & Capparella, 1992, due to the morphological similarities and wide distribution of its hosts in the Neotropical region. Therefore, this study encourages future taxonomic inquiries into I. similisi collected from other Saltator spp. in order to establish this synonymization of I. formarum with I. similisi, and hence, its wide distribution and dispersion in the Neotropical region, including across the Andes mountains.

Coccidia of Guinea fowls: Validity of recorded Eimeria spp. (Apicomplexa: Eimeriidae) and first molecular identification of Eimeria grenieri Yvoré & Aycardi, 1967.

Guinea fowls, Numida meleagris (L., 1758), are galliform birds native to sub-Saharan Africa, but introduced in several countries around the world for domestic breeding and/or animal production. This species is considered more resistant to disease by Eimeria spp. than other domestic galliform birds. Here we review the Eimeria spp. known to infect species of Numididae and provide the first molecular identification of an Eimeria sp. from Guinea fowls. There are currently 3 named eimerians from Guinea fowls; Eimeria numidae Pellerdy, 1962; Eimeria grenieri Yvoré and Aycardi, 1967; and Eimeria gorakhpuri Bhatia & Pande, 1967. We reviewed each of these species descriptions and documented their taxonomic shortcomings. From that, we suggest that E. gorakhpuri is a junior synonym of E. numidae. In conclusion, we have morphologically redescribed in detail E. grenieri from N. meleagris from Rio de Janeiro and provided molecular supplementation through sequencing of three non-overlapping loci in cox1 and cox3 genes and fragments of small and large subunit mitochondrial rDNA.

Isospora pichororei n. sp. (Chromista: Apicomplexa: Eimeriidae) from rufous-capped spinetails Synallaxis ruficapilla Vieillot, 1819 (Passeriformes: Furnariidae: Synallaxiinae) in South America.

Spinetails are a suboscine passerines of the genus Synallaxis Vieillot, 1818 which have great interest for ornithology, given the wide diversity of 37 species that are distributed throughout the Neotropical region. Despite this wide diversity and distribution, Synallaxis spp. have never been recorded as hosts of coccidian parasites. In this context, the current study describes a new species of Isospora Schneider, 1881 from rufous-capped spinetails Synallaxis ruficapilla Vieillot, 1819 captured in the Itatiaia National Park, which is a federal conservation unit in Southeastern Brazil. The oocysts of Isospora pichororei Genovez-Oliveira & Berto n. sp. are subspheroidal to ovoidal, measuring on average 25 by 21 µm. Micropyle is present, but discrete. Oocyst residuum absent, but one or two polar granules are present. Sporocysts are ellipsoidal with slightly pointed posterior end, measuring on average 17 by 10 µm. Stieda and sub-Stieda bodies are present. Sporocyst residuum is clustered among the vermiform sporozoites, which have striations, refractile bodies and nucleus. This morphology was different from the other Isospora spp. recorded in the host family Furnariidae. Molecular identification was targeted by the amplification and sequencing of a locus of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. This sequence had the highest similarity of 99.5% with a sequence deposited for Isospora oliveirai Ortúzar-Ferreira & Berto, 2020, which is a coccidian species that parasitizes suboscine tityrids Schiffornis virescens (Lafresnaye, 1838), also in the Itatiaia National Park. Phylogenetic analysis grouped some species in subclades, including I. pichororei with I. oliveirai; however, it was inconclusive in an expectation of parasite-host coevolution. Finally, I. pichororei is established as new to science, being the first description from Synallaxinae and the third description from Furnariidae. Furthermore, this is the first Isospora sp. from the host family Furnariidae to have a molecular supplementation by sequencing a locus of the cox1 gene of the mitochondrial genome.

Isospora juruviarae n. sp. (Apicomplexa: Eimeriidae) from chivi vireos Vireo chivi (Vieillot, 1817) (Passeriformes: Vireonidae) in South America.

Chivi vireos Vireo chivi (Vieillot, 1817) are passerine birds widely distributed throughout Brazil, but mainly observed in the Atlantic Forest of the South and Southeast regions of the country. In this context, the current study identifies a new species of Isospora Schneider, 1881 from V. chivi captured in the Marambaia Island, on the coast of the State of Rio de Janeiro, Southeastern Brazil. The oocysts of Isospora juruviarae Andrade & Berto n. sp. are subspheroidal to ovoidal, measuring on average 26 by 24 µm. Micropyle is absent or inconspicuous. Oocyst residuum absent, but polar granules are present. Sporocysts are ellipsoidal with pointed posterior end, measuring on average 17 by × 11 µm. Stieda and Sub-Stieda bodies are present. Sporocyst residuum is present among the vermiform sporozoites, which have refractile bodies and nucleus. This morphology was different from the other Isospora spp. recorded in the same family, superfamily and parvorder as the host. Molecular identification was targeted by the amplification and sequencing of two different loci of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and one locus of the 18S small subunit ribosomal RNA (18S) gene. Phylogenetic analyses were not very efficient in forming monophyletic groups associated with host taxon, zoogeographical region or taxonomic character; however, they confirmed the identification as a new species through comparison with sequences from Isospora spp. of wild passerines. Finally, based on the morphological and molecular analyses of the oocysts recovered from the chivi vireo V. chivi in the current work, I. juruviarae is considered new to science, being the second species recorded in the host family Vireonidae and the first to have a supplementation by molecular identification.

Molecular and statistical approaches to the delimitation of Eimeriidae species: a case of extreme polymorphism in eimerian oocysts from the plumbeous pigeon Patagioenas plumbea (Vieillot, 1818) (Columbiformes) in South America.

The current work aimed to analyze, morphologically, statistically, and molecularly, oocysts shed from plumbeous pigeons, Patagioenas plumbea (Vieillot, 1818), from a locality at 2197 m of altitude near the Agulhas Negras peak, the highest point of the State of Rio de Janeiro, southeastern Brazil. The oocysts were extremely polymorphic, being subspheroidal, ovoidal, or ellipsoidal, in addition to having the random presence/absence of characteristic features associated with the oocyst wall, such as micropyle, micropyle cap, lateral micropyle, and outer veil/rough wall. Linear regression confirmed the extreme polymorphism of oocysts, showing that if all combinations of taxonomic characters in oocysts (morphotypes) were overestimated, 19 different species could be identified/described. In contrast, the means comparison analysis between oocysts with the presence/absence of characteristic features and the histograms showed equivalences and regularity in the distribution in the classes of measures, which indicate the presence of a single species in the measured oocysts. Molecular analyses were performed from the isolation of individual oocysts of different morphotypes, which had their genetic material extracted, amplified, and sequenced in 4 non-overlapping loci in the cox1 and cox3 genes and fragments of the small and large subunit rDNA of mitochondrial DNA. The sequences were 100% identical between the morphotypes, with the exception of a very small divergence observed at the locus that partially covers the cox3 gene. The phylogenetic analysis was inconclusive for the locus within the cox1 gene traditionally used for eimeriid coccidians; however, the other loci should have a promising future for phylogenetic studies when more sequences for the same genic regions are deposited in GenBank. Finally, the multifactorial analysis of the current work supported that the polymorphic oocysts shed from P. plumbea are a single species, which was named Eimeria patagioenasae, making this the twenty-second eimerian description from Columbiformes.

Worldwide Dispersion of Coccidia from Migratory Birds: First Report of Eimeria bazi Chauhan et Bhatia, 1970 (Eimeriidae) Outside Asia from Buff-Necked Ibises Theristicus caudatus (Boddaert, 1783) (Threskiornithidae) in South America.

BACKGROUND: Eimeria spp. are coccidian protozoan parasites of domestic and wild animals. Pelecaniform birds are hosts of some Eimeria spp., however, from the family Threskiornithidae only one eimerian species is recorded, Eimeria bazi Chauhan et Bhatia, 1970 which was described from red-naped ibises Pseudibis papillosa (Temminck, 1824) in India. In this study, in turn, this species is morphologically and molecularly identified from buff-necked ibises Theristicus caudatus (Boddaert, 1783) in Brazil. PURPOSE: This study aimed to report E. bazi from buff-necked ibises T. caudatus in southeastern Brazil, revealing the worldwide distribution of this coccidian species, in addition to providing preliminary genotypic identification via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene. METHODS: A total of 73 fecal samples were collected from a flock of buff-necked ibises, which remained on the campus of the Federal Rural University of Rio de Janeiro (Universidade Federal Rural do Rio de Janeiro-UFRRJ) from March 2019 to August 2020. Fecal samples were processed by the Sheather's method to recover oocysts. The morphological and morphometrical studies of the oocysts were performed using an optical microscope and graphic editing software. Molecular analysis was performed by sequencing of the COI gene, and the phylogenetic analysis was based in the neighbor-joining and maximum likelihood estimates. RESULTS: Forty-five fecal samples were positive for oocysts identified as E. bazi. This oocysts are ovoidal, 26.2 × 18.9 µm, with smooth to slightly rough wall, c.1.7 µm thick. Micropyle robust and protruding, sometimes with a polar body attached. Oocyst residuum absent, but one or two small polar granules are present. Sporocysts ovoidal to lemon-shaped, 14.2 × 8.7 µm. The Stieda body is knob-like to rounded and sub-Stieda body is absent or vestigial. Sporocyst residuum is composed of granules often membrane-bound. Sporozoites are vermiform, with refractile bodies. This morphology was consistent with the original description of E. bazi from P. papillosa in India. Molecular analysis at the COI gene exhibited low similarity with coccidians sequenced for the same genic region deposited in GenBank, sitting E. bazi separately on the cladogram. CONCLUSIONS: The morphological and molecular studies support the identification of E. bazi from T. caudatus in South America, thus revealing the wide distribution of this eimerian species in the world provided by migratory birds and/or with intercontinental distribution.

Territorial expansion of Zenaida auriculata (Aves: Columbidae) in the Brazilian Amazon / Expansão territorial de Zenaida auriculata (Aves: Columbidae) na Amazônia brasileira

Birds are considered bioindicators of environmental quality because they are sensitive to environmental changes. The eared dove (Zenaida auriculata) is associated with open areas, has low sensitivity to environmental changes and is widely distributed in Brazil, although it is not typically found in the Amazon region. We analyzed the biogeographic expansion of this species in the Brazilian Amazon through searches carried out on digital citizen science platforms. The records of Z. auriculata were related with the area of accumulated deforestation in the biome. We identified 804 records of Z. auriculata in the Brazilian Amazon, of which 259 were in areas where its occurrence had been previously unknown. We show that the species has been expanding its geographic distribution, clearly overlapping with deforested areas.(AU)

As aves são consideradas bioindicadores de qualidade ambiental por serem sensíveis às mudanças ambientais. A avoante (Zenaida auriculata) é associada a áreas abertas, possui baixa sensibilidade às mudanças ambientais e é amplamente distribuída no Brasil, apesar de não ser tipicamente encontrada na Amazônia. Nós analisamos a expansão biogeográfica desta espécie na Amazônia brasileira por meio de buscas em plataformas digitais de ciência cidadã. Os registros obtidos de Z. auriculata foram relacionados com a área de desmatamento acumulado no bioma. Identificamos 804 registros de Z. auriculata na Amazônia brasileira, dos quais 259 em regiões onde sua ocorrência era desconhecida. Nós mostramos que a espécie vem ampliando sua distribuição geográfica em nítida sobreposição com áreas desmatadas.(AU)

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.

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.