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.

Morphological and molecular biology identification of Cystoisospora sp. in the blue fox, Alopex lagopus (Linnaeus, 1758).

To investigate the prevalence and molecular characteristics of Cystoisospora sp. in blue fox (Alopex lagopus), Sheather's sugar floatation method was conducted to detect coccidia in 423 fresh fecal samples randomly collected from blue fox farms from three cities in China. The overall prevalence of coccidia was 1.4% (6/423), and three Cystoisospora sp. (Cystoisospora fennechi, Cystoisospora sp. I and Cystoisospora vulpina) were identified by their morphological characteristics. The 18S ribosomal RNA (rRNA) and cytochrome c oxidase subunit I (COI) locus sequences were sequenced for molecular biological identification, homology comparison, and phylogenetic analysis of Cystoisospora sp. by single-oocyst selection technology and multi-locus-nested PCR amplification. At the 18S rRNA and COI loci, C. vulpina had 99.48% and 99.59% homology, respectively, with Cystoisospora canis and Cystoisospora ohioensis from canines. Phylogenetic analysis indicated that C. vulpina was clustered in a clade with Cystoisospora sp. from Canidae, which the relatives are consistent with the hosts. To our knowledge, this is the first report on molecular identification and evolutionary analysis of C. vulpina at two different loci.

Molecular identification of Isospora coerebae Berto, Flausino, Luz, Ferreira amp; Lopes, 2010 (Chromista: Miozoa: Eimeriidae) from the bananaquit Coereba flaveola (Linnaeus, 1758) (Passeriformes: Thraupidae: Coerebinae) from Brazil.

Isospora coerebae Berto, Flausino, Luz, Ferreira Lopes, 2010 is a coccidian protozoan described from bananaquits Coereba flaveola (Linnaeus, 1758), on Marambaia Island, which is located on the southeastern Brazilian coast. In this current work, I. coerebae is identified from C. flaveola in a protected area close to Marambaia Island, but on the mainland, establishing a new location of parasitism, in addition to providing a preliminary genotypic characterization via sequencing of two regions of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene. Its oocysts are sub-spherical, 24.4 22.9 m, with smooth, bilayered wall, ~1.7 m thick. Micropyle, polar granules and oocyst residuum are absent. Sporocysts are elongate ovoidal, 17.6 10.5 m. Stieda body prominent and rounded and sub-Stieda body short and wide. Sporocyst residuum is composed of scattered granules of different sizes. Sporozoites are vermiform with a prominent posterior refractile body. The oocysts of the current work are morphologically equivalent to the original description of I. coerebae, which have a typical and easily identifiable morphology, mainly in the Stieda and Sub-Stieda bodies. The two sequenced gene regions of the COI gene approximated I. coerebae to Isospora spp. from Southeastern Brazil, but also from Isospora spp. from passerines of North America, Europe and Asia. Although there is a small overlap between the two genic regions sequenced in the current work, it is estimated that the longer COI sequence, which was recently designed and still not widespread, should show better phylogenetic results in the future.

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.

The transcriptome from asexual to sexual in vitro development of Cystoisospora suis (Apicomplexa: Coccidia).

The apicomplexan parasite Cystoisospora suis is an enteropathogen of suckling piglets with woldwide distribution. As with all coccidian parasites, its lifecycle is characterized by asexual multiplication followed by sexual development with two morphologically distinct cell types that presumably fuse to form a zygote from which the oocyst arises. However, knowledge of the sexual development of C. suis is still limited. To complement previous in vitro studies, we analysed transcriptional profiles at three different time points of development (corresponding to asexual, immature and mature sexual stages) in vitro via RNASeq. Overall, transcription of genes encoding proteins with important roles in gametes biology, oocyst wall biosynthesis, DNA replication and axonema formation as well as proteins with important roles in merozoite biology was identified. A homologue of an oocyst wall tyrosine rich protein of Toxoplasma gondii was expressed in macrogametes and oocysts of C. suis. We evaluated inhibition of sexual development in a host-free culture for C. suis by antiserum specific to this protein to evaluate whether it could be exploited as a candidate for control strategies against C. suis. Based on these data, targets can be defined for future strategies to interrupt parasite transmission during sexual development.

Prevalence and genetic characteristics of Blastocystis hominis and Cystoisospora belli in HIV/AIDS patients in Guangxi Zhuang Autonomous Region, China.

Blastocystis hominis and Cystoisospora belli are considered to be common opportunistic intestinal protozoa in HIV/AIDS patients. In order to investigate the prevalence and genetic characteristics of B. hominis and C. belli in HIV/AIDS patients, a total of 285 faecal samples were individually collected from HIV/AIDS patients in Guangxi, China. B. hominis and C. belli were investigated by amplifying the barcode region of the SSU rRNA gene and the internal transcribed spacer 1 (ITS-1) region of the rRNA gene, respectively. Chi-square test or Fisher's exact test were conducted to assess the risk factors related to B. hominis and C. belli infection. The prevalence of B. hominis and C. belli was 6.0% (17/285) and 1.1% (3/285) respectively. Four genotypes of B. hominis were detected, with ST3 (n = 8) and ST1 (n = 6) being predominant, followed by ST6 (n = 2) and ST7 (n = 1). Females had a statistically higher prevalence of B. hominis (11.6%) than males (4.2%). The statistical analysis also showed that the prevalence of B. hominis was significantly associated with age group and educational level. Our study provides convincing evidence for the genetic diversity of B. hominis, which indicates its potential zoonotic transmission and is the first report on the molecular characteristics of C. belli in HIV/AIDS patients in China.

Progression of asexual to sexual stages of Cystoisospora suis in a host cell-free environment as a model for Coccidia.

Coccidia display a characteristic life cycle, where the parasites switch between asexual and sexual development, resulting in an environmental stage, the oocyst. The entero-pathogenic Cystoisospora suis, a coccidian parasite of swine and close relative to Toxoplasma gondii, undergoes development in one host-cycle. Despite the well-described intracellular development of Coccidia, the C. suis life cycle can progress in an in vitro, host cell-free system after initial intracellular development of merozoites. A novel host cell-free cultivation method was developed by transferring purified merozoites from cell culture supernatant (dpi 6) to culture medium and incubating them for 5 days to induce their progression to sexually differentiated stages. The development of sexual stages in the absence of host cells was verified by morphological studies, flow cytometry and the transcription analysis of three genes linked to sexual stages (HAP2, OWP and TyRP). The host cell-free culture permits the sexual development (and with this, the complete life cycle progression from sporozoites to oocysts) of C. suis in vitro and provides a new tool for detailed research on the development of C. suis and possibly other Coccidia. This will also be useful for the evaluation of novel drug or vaccine targets in these parasites.

Phylogenetic characterization of Isospora jaracimrmani oocysts from a veiled chameleon (family Chamaeleonidae; Chamaeleo calyptratus) reared at a zoo in Ishikawa, Japan.

Oocysts of Isospora sp. were detected in the feces of a veiled chameleon (family Chamaeleonidae; Chamaeleo calyptratus) kept at a zoo in Ishikawa, Japan. Phylogenetic analysis placed the sequence in the cluster of Isospora spp. isolated from reptiles. Based on a comparison of morphological data of ten previously reported Isospora species from the Chamaeleonidae family, this isolate was morphologically similar to I. jaracimrmani, which has been considered to be a virulent species. This case study suggests the possibility that species of Isospora might not always cause disease because the animal that shed these oocysts showed no symptoms for more than two months.

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.

Morphological and molecular characterization of Isospora amphiboluri (Apicomplexa: Eimeriidae), a coccidian parasite, in a central netted dragon (Ctenophorus nuchalis) (De Vis, 1884) in Australia.

An Isospora species, Isospora amphiboluri, originally described by Canon in 1967 and later by McAllister et al. (1995), was isolated from a central netted dragon (Ctenophorus nuchalis) housed at a wildlife rehabilitation centre in Perth, Western Australia. Sporulated oocysts of Isospora amphiboluri (n = 30) are spherical, 24.2 (26.5-23.0) µm in length and 23.9 (22.4-25.9) µm in width, with a shape index of 1.01. The bilayered oocyst wall is smooth and light-yellow in color. Polar granule, oocyst residuum and micropyle are absent. The sporocysts are lemon-shaped, 15.7 (15.2-18.0) × 10.2 (8.9-11.2) µm, with a shape index (length/width) of 1.53. Stieda and substieda bodies are present, the Stieda body being small and hemidome-shaped and the substieda half-moon-shaped. Each sporocyst contains four vermiform sporozoites arranged head to tail. The sporozoites are 11.7 (9.9-16.2) × 3.0 (2.4-3.5) µm, with a shape index (length/width) of 3.87. A sporocyst residuum is present. Sporozoites contain a central nucleus with a finely distributed granular residuum. Comparison of oocyst measurements and their features with other valid Isospora species from hosts in the Agamid family confirmed that this Isospora species is Isospora amphiboluri. Molecular characterization of I. amphiboluri at the 18S rRNA and MTCOI loci showed the highest similarity with I. amphiboluri from the central bearded dragon, 99.8% and 99.7% respectively. This is the first report of I. amphiboluri from a central netted dragon in Australia.

Distribution, redescription, and molecular identification of Isospora striata McQuistion et al. 1997 (Eimeriidae), from woodcreepers (Dendrocolaptidae) in South America.

Woodcreepers are passerines of the family Dendrocolaptidae, which have a high forest dependency. The current work aimed to redescribe Isospora striata McQuistion et al. 1997, from two new hosts in protected areas in Brazil, revealing new localities of parasitism, in addition to providing preliminary genotypic identifications via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene from both host species. Isospora striata has oocysts that are subspheroidal to ovoidal, 19.4 × 16.8 µm with smooth wall. Oocyst residuum is absent, but micropyle and polar granules are present. Sporocysts are ovoidal, 13.6 × 8.3 µm, with both Stieda and sub-Stieda bodies. Sporocyst residuum is present and sporozoites with refractile body, nucleus, and striations. The morphological study and the 100% similarity in sequencing of the COI gene between samples of different dendrocolaptid species confirmed the identification of a single species, supporting the identification of I. striata in the Brazilian Atlantic forest and consequently the wide distribution of this coccidian species in the Neotropical Region.

Morphological and molecular description of a new species of Isospora (Apicomplexa) from a New Holland honeyeater (Phylidonyris novaehollandiae).

A new Isospora species is described from New Holland honeyeaters (Phylidonyris novaehollandiae). Sporulated oocysts (n = 25) were characterised as subspheroidal, 29-32 × 28-31 (29.8 × 29.4); length/width (L/W) ratio 1.01-1.02 (1.01). Wall bi-layered, 1.3-1.6 (1.5) thick, outer layer smooth, c.2/3 of total thickness. Micropyle and oocyst residuum absent, but usually two polar granules are present. Sporocysts (n = 25) ovoidal, 18-19 × 12-14 (18.4 × 12.3); L/W ratio 1.42-1.53 (1.50). Stieda body present, flattened, c.0.5 deep × 2.5 wide; sub-Stieda present, rounded, c.2.5 deep × 3.5 wide; para-Stieda body absent; sporocyst residuum present, usually a distinctly irregular body consisting of numerous small granules that appear to be membrane-bound. Sporozoites vermiform, with robust anterior and posterior refractile bodies. Molecular characterization was conducted at the 18S and 28S ribosomal RNA and the mitochondrial (mt) cytochrome oxidase (COI) loci. Phylogenetic analysis of genomic 18S and mt COI sequences indicated that Isospora phylidonyrisae n. sp. was genetically similar to Isospora coronoideae, isolated from an Australian raven (Corvus coronoides) in Western Australia, with a 99.3% and 98.4% homology, respectively. The 28S rRNA sequence was most similar to Isospora anthochaerae (KF766053) and Isospora manorinae (KT224381), both with a 98.2% genetic similarity. Based on morphological and genetic data, this isolate is a new species of Isospora, which is named Isospora phylidonyrisae n. sp. after its host.

Morphological and genetic characterization of the first Isospora species (I. lugensae n. sp.) from a Kerguelen petrel (Lugensa brevirostris).

A new coccidian species, Isospora lugensae n. sp., was described from a single Kerguelen petrel (Lugensa brevirostris). Sporulated oocysts (n = 25) were characterized as subspheroidal to ellipsoidal measuring 24-25 µm × 21-23 µm (24.8 × 22.2 µm) in length/width (L/W), respectively, with a ratio of 1.07-1.14 µm (1.12). They contained a bi-layered wall with a thickness of 0.8-1.2 µm (1.0) and the outer layer smooth, with c.2/3 of total thickness. The oocyst contained two polar granules with both micropyle and oocyst residuum absent. Ovoidal sporocysts (n = 25) measured 15-16 µm × 10-11 µm (15.7 × 10.8 µm) in L/W, with a ratio of 1.41-1.49 µm (1.46). A flattened to knob-like Stieda body was present (c.0.5 µm deep × 2.5 µm wide) as well as a rounded to trapezoidal sub-Stieda (c.1.5 µm deep × 3.0 µm wide); however, no para-Stieda body was detected. The sporocyst residuum was composed of scattered spherules of different sizes, while vermiform sporozoites contained a refractile body, nucleus and visible striations. Analysis of the full-length mitochrondrial (mtDNA) genome revealed 3 protein-coding genes, (CytB, COI and COIII), 18 LSU and 14 small subunit (SSU) rDNA fragments, without transfer RNA genes with a total length of 6257 bp. Phylogenetic analysis of genomic SSU ribosomal sequences indicated that Isospora lugensae n. sp. is genetically similar to Eimeria reichenowi, isolated from a red-crowned crane (Grus japonensis) from Japan, with a 96.6% homology. The mtDNA sequence is most similar to Isospora serinuse with a 95.8% genetic similarity. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that to date has only been found in a Kerguelen petrel.

Parasites in a hotspot: diversity and specificity patterns of apicomplexans infecting reptiles from the Socotra Archipelago.

Although parasites represent a major component of biodiversity, they remain poorly assessed, especially in remote regions. In this study, we screened 461 reptiles from Socotra, the largest and most biologically diverse archipelago in Arabia. Using 18S rRNA primers, we detected various apicomplexan parasites, namely haemogregarines, sarcocystids and eimeriids. Haemogregarines were the most common and genetically diverse, followed by sarcocystids (genus Sarcocystis) and eimeriids (genera Isospora and Lankesterella). All were related to parasites of other reptiles, including species from Arabia, Northern Africa and Asia. Like their 29 endemic reptile hosts, almost all Socotran parasites presented high genetic divergence and ecological differences from those found elsewhere, and probably represent undescribed endemic species. Among hosts, skinks were the most parasitized, which contrasted with similar studies from other areas, probably due to their more generalist diet and habitat use. As expected due to its high species richness, geckos harboured the highest parasite diversity in the archipelago. Parasite diversity also seemed to be correlated to island size, as the largest island harboured most haplotypes. This study emphasizes the importance of screening parasites in wild hosts from remote regions and of considering host ecology to understand disease transmission across taxa.

Pitfalls in Morphologic Diagnosis of Pathogens: Lessons Learned From the Pseudo-Cystoisospora Epidemic.

Multiple groups have recently reported involvement of the gallbladder mucosa of immunocompetent patients by cystoisospora organisms. However, this has recently been disproved with the support of molecular and ultrastructural studies. Here we present a summary of these events, recounting how this pseudo-Cystoisospora epidemic began and ended. This review also highlights the important role played by ancillary techniques in supplementing the morphologic diagnosis of pathogens.

Detection and characterization of the Isospora lunaris infection from different finch hosts in southern Iran.

This study was conducted to investigate the Isosporoid protozoan infections in finch types. Fecal samples were collected from marketed domestic Java sparrows (Lonchura oryzivora), colored and white Zebra finch (Taeniopygia guttata), and European goldfinch (Carduelis carduelis) in southern Iran. The coccidial oocysts were recovered and investigated according to the morphological features and the ribosomal gene markers. Additionally, a challenge infection was conducted with 5 × 104 and 5 × 103 sporulated oocysts in four java sparrows to estimate the clinical manifestations. Based on the morphology, the oocysts of Isospora lunaris were identified in all sampled bird types; however, the molecular method revealed the isolates had considerable similarities with some of Isospora and systemic Isospora-like organisms named as Atoxoplasma. Phylogenetic data also constructed an Atoxoplasma/Isospora clade with high sequence identities. High dose of the challenge with the parasite led to severe depression and sudden death, but it did not coincide with remarkable lesions and parasitic invasion in visceral organs. Contrary to molecular results, this feature is consistent with the common Isospora infections in passerines and differs from those described for Atoxoplasma species. Because of the prevalence, possibility of transmission, and clinical consequences, preventive measures are necessary to avoid outbreaks of isosporoid infections among finch type birds.

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.

Morphological and molecular characterization of Cystoisospora laidlawi oocysts (Apicomplexa: Eimeriidae) in farmed American mink (Neovison vison) in Denmark.

From a longitudinal survey conducted on 30 Danish mink farms in 2016, 11.0% of faecal samples (456/4140) were positive for Cystoisospora laidlawi oocysts by microscopy, with 60% (189/315) of mink being positive at least once during the study period. Morphological analysis of sporulated oocysts identified Cystoisospora oocysts measuring 34.3 × 29.5 µm with an oocyst length/width (L/W) ratio of 1.2. The morphological features of the oocysts were identical to Isospora laidlawi previously morphological identified in farmed mink from Denmark and elsewhere. Phylogenetic analysis of 18S rDNA sequences (1221 bp) from three positive mink indicated that Cystoisospora from mink shared the highest genetic similarity to C. canis from a Canadian dog (99.6%). The phylogenetic analysis placed Cystoisospora from mink in a clade with other Cystoisospora isolates.

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.