On the validity and distribution of Myxobolus curemae Vieira, Agostinho, Negrelli, Silva, Azevedo and Abdallah, 2022 (Myxozoa: Bivalvulida) from a new record on the coast of southeastern Brazil.

Myxobolus spp. are parasites of wide diversity and distribution in fish, both in the natural environment and in freshwater or marine farming systems around the world and are sometimes associated with severe disease in their hosts (Schmahl et al. 1989; Lom and Dyková, 1995; Eiras et al. 2021). The white mullet Mugil curema Valenciennes, 1836 is a pelagic fish of the Mugilidae family widely distributed in the Atlantic Ocean (Froese & Pauly 2022). Recently, Myxobolus curemae Vieira, Agostinho, Negrelli, Silva, Azevedo and Abdallah, 2022 was described from white mullets Mugil curema Valenciennes, 1836 in Brazil (Vieira et al. 2022). In the current work, Myxobolus sp. was identified with the same biological and morphological characteristic features of M. curemae from white mullets from the Jacarepaguá Lagoon Complex in the State of Rio de Janeiro, Brazil, in addition to being 100% identical for a genic region of the nuclear small subunit ribosomal DNA. However, this Myxobolus sp. was also similar to Myxobolus hani Faye, Kpatcha, Diebakate, Fall and Toguebaye, 1999, which was described from white mullets in Senegal, Africa, without molecular identification (Faye et al. 1999). Therefore, this current work makes observations on these Myxobolus spp. which are morphologically and biologically similar, and that could establish M. curemae as a junior synonymy of M. hani.

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.

Isospora leptopogoni n. sp. (Apicomplexa: Eimeriidae) from the sepia-capped flycatcher Leptopogon amaurocephalus Tschudi, 1846 (Passeriformes: Rhynchocyclidae) in South America.

Coccidian protozoan species recorded from flycatchers are few, but they have been described with a certain frequency in recent years. In this context, the present study describes a new Isospora sp. from sepia-capped flycatchers Leptopogon amaurocephalus Tschudi, 1846 captured in the Itatiaia National Park and in a reforestation area which is about 60 km away from the park boundaries, in addition to providing a molecular identification via sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene. Isospora leptopogoni n. sp. has oöcysts that are subspheroidal to ovoidal, measuring on average 22.0 × 19.7 µm, with a smooth, bi-layered wall, c.1.7 µm thick. The micropyle is delicate or inconspicuous. Oöcyst residuum is absent, but one to three polar granules are present. Sporocysts are lemon-shaped, measuring on average 14.7 × 9.3 µm, with a knob-like Stieda body and a rectangular to rounded sub-Stieda body. Sporocyst residuum is present, consisting of compactly bounded granules. Sporozoites are vermiform, with refractile bodies and nucleus. Isospora leptopogoni is different from other Isospora spp. mainly due to its lemon-shaped sporocysts, the presence of micropyle and details of Stieda and sub-Stieda bodies. Phylogenetic analysis placed I. leptopogoni close to other Isospora spp. recorded from phylogenetically related hosts and from the same biogeographic region. Finally, the recurrent finding of this coccidian species in the same L. amaurocephalus specimen in a specific locality in the Itatiaia National Park suggests that the dispersion of I. leptopogoni needs continuous transmissions between susceptible passerines as the area of movement of each L. amaurocephalus specimen appears to be quite small.

Observations on an Eimeria sp. (Apicomplexa) from the green kingfisher Chloroceryle americana (Gmelin, 1788) (Coraciiformes) in Southeastern Brazil: an example of how the ecological aspects of the host can be essential for the identification of its coccidians.

Parasitism in kingfishers is very little reported and predominantly related to hemoparasites, helminths, and ectoparasites. The present study provided a morphological and genotypic study of an Eimeria sp. recovered from a green kingfisher Chloroceryle americana (Gmelin, 1788) captured in the Marambaia Island, on the coast of the state of Rio de Janeiro, Southeastern Brazil. The coccidial density, some morphological aspects of its oocysts, the molecular results, and, mainly, the ecological niche of C. americana in the mangrove of the Marambaia Island suggest that this coccidian species is a pseudoparasite.

Isospora spp. (Eimeriidae) from green-winged saltators Saltator similis d'Orbigny & Lafresnaye, 1837 (Thraupidae) from captivity near the Conservation Unit of the Itatiaia National Park in Southeastern Brazil.

The present study identifies three Isospora spp. recorded from faecal samples of green-winged saltators Saltator similis d'Orbigny & Lafresnaye kept in captivity in the surroundings of the Itatiaia National Park, which is a Conservation Unit with a high degree of vulnerability located in the Serra da Mantiqueira on the border of the States of Rio de Janeiro and Minas Gerais. Isospora saltatori Berto, Balthazar, Flausino & Lopes, 2008, Isospora trincaferri Berto, Balthazar, Flausino & Lopes, 2008 and Isospora similisi Coelho, Berto, Neves, Oliveira, Flausino & Lopes, 2013 were compatible in all characteristic features with their respective original descriptions, despite some divergences that are discussed in this study. In addition to the preliminary morphological identification, this study provided a preliminary genotypic identification of these three Isospora spp. via sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene, which was suitable for the genotypic differentiation of these three coccidians, but was inconclusive in the phylogenetic analysis. Finally, this study discusses the environmental risks of these coccidians in captivity of green-winged saltators in the surroundings of the Itatiaia National Park.

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.

Polymorphism and genetic diversity of Isospora parnaitatiaiensis Silva, Rodrigues, Lopes, Berto, Luz, Ferreira & Lopes, 2015 (Eimeriidae) from antbirds (Thamnophilidae) in Brazil.

Isospora parnaitatiaiensis Silva, Rodrigues, Lopes, Berto, Luz, Ferreira & Lopes, 2015 was identified from a new host, the plain antvireo Dysithamnus mentalis (Temminck), and also from the white-shouldered fire-eye Pyriglena leucoptera Vieillot, in its type-locality, the Itatiaia National Park in the southeastern Brazil, and a preliminary genotypic characterisation by sequencing the mitochondrial cytochrome c oxidase subunit 1 gene is provided. The oöcysts recovered from P. leucoptera and D. mentalis were polymorphic and have genotypic differences that were not considered sufficient for the description of new species, but only different genotypes and morphotypes of I. parnaitatiaiensis related to each host. These morphological and molecular variations were associated with a process of ongoing speciation and in adaptive development to their respective host species.

First report of Myxobolus episquamalis Egusa, Maeno & Sorimachi, 1990 (Myxozoa: Bivalvulida) in Lebranche mullet Mugil liza Valenciennes, 1836 (Teleostei: Mugiliformes) from Neotropical region.

In the current study, Myxobolus episquamalis Egusa, Maeno & Sorimachi, 1990 (Myxozoa: Bivalvulida) is reported from the Lebranche mullet Mugil liza Valenciennes, 1836 in the estuarine region of the Maricá Lagoon, State of the Rio de Janeiro, southeastern Brazil. To date, this myxozoan species was reported in mullets from Asia, Africa, Europe, and Oceania. The characteristics of M. episquamalis previously reported are similar to the findings of the present study. DNA sequences of the nuclear small subunit ribosomal DNA (SSU rDNA) had 99.7-100% similarity with the sequences of M. episquamalis from North Africa and Asia. Therefore, strong morphological and molecular similarities ensure the identification of M. episquamalis in the current study. Finally, this finding records a new host and locality, revealing the worldwide distribution of this myxozoan species.

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.

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.

The vulnerable Sporophila frontalis (Verreaux) and Haplospiza unicolor Cabanis as new hosts for Isospora sporophilae Carvalho-Filho, Meireles, Ribeiro & Lopes, 2005 (Eimeriidae) in Brazil.

Isospora sporophilae Carvalho-Filho, Meireles, Ribeiro & Lopes, 2005 was morphologically and molecularly identified from the double-collared seedeater Sporophila frontalis (Verreaux), which is categorised as 'vulnerable' by the International Union for Conservation of Nature and Natural Resources (IUCN), and from the uniform finch Haplospiza unicolor Cabanis in conserved and anthropomorphic/fragmented areas of Atlantic Forest in the southeastern Brazil. The oöcysts recovered from S. frontalis and H. unicolor had small morphological and genotypic differences that were not considered sufficient for the description of new species, but only different genotypes of I. sporophilae related to each host. This coccidian species was originally described from double-collared seedeaters Sporophila caerulescens (Vieillot) in a center screening of wild animals; therefore, this new report emphasises a potential occurrence of anthropomorphic dispersion of coccidia through illegal trade, seizures and reintroductions in the wild.

Isospora lopesi n. sp. (Apicomplexa: Eimeriidae) from the eastern white-throated spadebill Platyrinchus mystaceus Vieillot (Passeriformes: Tyranni: Tyrannidae) in South America.

A species of Isospora Schneider, 1881 (Protozoa: Apicomplexa: Eimeriidae) considered as new to science is described and characterised molecularly from the eastern white-throated spadebill Platyrinchus mystaceus Vieillot in the Parque Nacional do Itatiaia, southeastern Brazil. Isospora lopesi n. sp. has oöcysts that are subspheroidal to ovoidal, 18-24 × 18-22 (20.6 × 19.7) µm, with smooth, bilayered wall, c.1.5 µm thick. Micropyle and oöcyst residuum are absent, but one polar granule is present. Sporocysts are ellipsoidal, 12-16 × 8-11 (14.4 × 8.6) µm. The Stieda body is flattened to half-moon-shaped and sub-Stieda body is rounded. Sporocyst residuum is present, consisting of numerous spherules of different sizes. Sporozoites are vermiform with anterior and posterior refractile bodies 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 spectacled warblers Sylvia conspicillata Temminck, 1820. This is the fourth isosporoid coccidian described from New World tyrannid birds, but is the first to have a complementary molecular characterisation.