Immature ticks on wild birds and the molecular detection of a novel Rickettsia strain in the Ibitipoca State Park, southeastern Brazil.

Birds are recognized hosts of ticks, especially for the immature stages which may harbor various species and strains of Rickettsia. To explore landscapes inhabited by birds and their ticks would expand the knowledge on host-parasite relationships and the rickettsiae. The aim of this paper was to record the diversity of ticks collected on wild birds and assess the phylogenetic position of a novel Rickettsia strain detected in immature ticks. Birds were captured in the Ibitipoca State Park, located in the Minas Gerais state, southeastern Brazil, as part of a long-term research project on the ecology of ticks, birds and Rickettsia. We found three tick species parasitizing birds: Amblyomma aureolatum (63 larvae, 10 nymphs), Haemaphysalis leporispalustris (28 larvae, seven nymphs) and Amblyomma romarioi (27 larvae). Among these, A. aureolatum was the most abundant species including 54% (73/135) of the collected ticks. New tick-host records were: A. romarioi on Turdus amaurochalinus and H. leporispalustris on Thamnophilus caerulescens, Saltator similis and Zonotrichia capensis. Of the 82 ticks tested for Rickettsia spp. by PCR, two larvae (2.5%) of A. romarioi were infected with 'Candidatus Rickettsia paranaensis', a novel putative Rickettsia species closely related to Rickettsia africae, Rickettsia sibirica and Rickettsia parkeri, as corroborated by our phylogenetic analysis. Finally, we present a list of all records of immature stages of H. leporispalustris on passerine birds in Brazil.

Ticks (Acari: Ixodidae) and tick-borne agents associated with domestic dogs in an environmental protection area in Brazil, with molecular evidence of Rhipicephalus linnaei (Audouin, 1826).

Wild animals and domestic dogs living in human dwellings near forested areas can share ectoparasites, including ticks. In this study, we surveyed ticks associated with dogs which tutors living in the Palmares Environmental Protection Area (EPA Palmares). Dogs were classified into three categories, domiciled, semi-domiciled and wandering dogs according to dog care/ type of dwelling. Ticks were collected monthly from January to December, 2020. Overall, 60 (33.9%) out of 177 examined dogs were infested by ticks. Six species of ticks were identified: Rhipicephalus linnaei, Amblyomma aureolatum, Amblyomma sculptum, Amblyomma ovale, Amblyomma dubitatum and Rhipicephalus microplus. The overall prevalence and presence in semi-domicilied+wandering dogs was higher for A. aureolatum than for R. linnaei by the Chi-square statistic tests. A random sample of 50 ticks, collected from 22 different dogs, were processed through molecular analyses. Ticks were submitted to DNA extraction and also by PCR, using specific primers in order to pathogens monitoring. Four males of A. aureolatum yielded DNA sequences (350 bp) that were 100% identical to the type strain of Rickettsia bellii in GenBank (CP000087).

A new species of Caryospora Léger, 1904 (Apicomplexa: Eimeriidae) from the snake Philodryas olfersii Lichtenstein (Colubridae) from a coastal habitat in Brazil.

A new coccidian species of the genus Caryospora Léger, 1904 (Protozoa, Apicomplexa, Eimeriidae) is reported from the colubrid snake host Philodryas olfersii Lichtenstein at a coastal area in the State of Rio de Janeiro, south-eastern Brazil. Oöcysts of Caryospora olfersii n. sp. are spherical to sub-spherical, 33.1 × 31.2 µm, with smooth, colourless, three-layered wall, c.1.4; middle layer lightly striated. Micropyle, oöcyst residuum and polar granule are all absent. Sporocysts are ovoid, 22.9 × 17.4 µm on average, with one extremity in the shape of a short neck. Stieda body present, 3.2 × 1.3 µm, sub-Stieda body present, homogeneous, larger than Stieda body, 4.5 × 1.7 µm. Sporozoites are inserted in a bulky sporocyst residuum.

Isospora canaria Box, 1975 (Apicomplexa: Eimeriidae) from canaries Serinus canaria Linnaeus (Passeriformes: Fringillidae) in Brazil.

Isospora canaria Box, 1975 (Protozoa, Apicomplexa, Eimeriidae) is reported and described from canaries Serinus canaria (Linnaeus) in southeast Brazil. Its oöcysts are subspheroidal to ellipsoidal, 24.4 × 22.2 µm, with smooth, bilayered wall, ~1.2 µm. The micropyle and the oöcyst residuum are absent, but a polar granule is present. The sporocysts are lemon-shaped, 17.6 × 10.6 µm. The Stieda body is nipple-like, and substieda body is prominent and homogeneous. The sporocyst residuum is composed of scattered granules. The sporozoite has anterior and posterior refractile bodies and a nucleus. The report of this coccidium recovered from exotic canaries in South America is relevant to native passerines, mainly to Carduelis yarrellii, which are listed as vulnerable species by IUCN (International Union for Conservation of Nature and Natural Resources).

Isolation and characterization of Cyniclomyces guttulatus (Robin) Van Der Walt and Scott, 1971 in dogs in Brazil.

Vegetative cells of an ascomycetous yeast, morphologically consistent with published descriptions of Cyniclomyces guttulatus, were observed in large numbers in the feces and stomach washes of three dogs with a recurrent medical history characterized by vomiting and diarrhea. Nucleotide sequence analysis of an approximately 600 base pair fragment of the variable D1/D2 domain of large subunit (26S) ribosomal DNA of a pure culture, isolated from a Siberian Husky, revealed 98-99 % homology to sequences deposited in the GenBank as C. guttulatus. These data represent the first observation of C. guttulatus in association with canine gastrointestinal illness in the southern hemisphere and add weight to the hypothesis that this yeast may act as an opportunistic pathogen of dogs. An extended examination of wet mounts and smears prepared from feces collected from 63 dogs with no clinical symptoms of gastrointestinal illness, identified C. gluttulatus in 14 (22.2 %) of the animals, albeit at lower numbers than in diseased dogs, indicating that this yeast species is widely distributed as a component of the normal microflora of the canine gastrointestinal tract.

Isospora bocamontensis n. sp. (Apicomplexa: Eimeriidae) from the yellow cardinal Gubernatrix cristata (Vieillot) (Passeriformes: Emberizidae) in South America.

A new coccidian species (Protozoa: Apicomplexa: Eimeriidae) is reported from the endangered yellow cardinal Gubernatrix cristata (Vieillot) in southern Brazil. Isospora bocamontensis n. sp. has oöcysts which are subspheroidal, measure 32.1 × 28.9 µm and have a smooth, bilayered wall c.1.5 µm thick. The micropyle and the oöcyst residuum are absent, but a polar granule is sometimes present. Its sporocysts are ellipsoidal and 17.3 × 12.2 µm in size and contain a half-moon-shaped Stieda body, a prominent, homogeneous substieda body; and a sporocyst residuum composed of a compact mass of granules. The sporozoites have one refractile body and a nucleus.

Coccidia of New World passerine birds (Aves: Passeriformes): a review of Eimeria Schneider, 1875 and Isospora Schneider, 1881 (Apicomplexa: Eimeriidae).

In the New World, the avian order Passeriformes comprises 47 families and 2,453 species, yet to date only 21 (45%) of the families and 58 (2%) of the species have been examined for coccidia, and from these only two species of Eimeria Schneider, 1875 and 81 species of Isospora Schneider, 1881 have been described. This review contributes to our understanding of the morphology and systematics of coccidian parasites of passeriforms, providing a scientific basis for the identification of sporulated oöcysts recovered from the faeces of passerine birds from North, Central and South America. To this end, the coccidia were organised and grouped according to the family of the host, following the widely recognised concept of family-specificity and the updated systematics of the class Aves. Details of 83 eimeriid species are presented along with an illustration and tabulated data.

Isospora piacobrai n. sp. (Apicomplexa: Eimeriidae) from the masked yellowthroat Geothlypis aequinoctialis (Gmelin) (Passeriformes: Parulidae) in South America.

A new coccidian species (Protozoa: Apicomplexa: Eimeriidae) obtained from masked yellowthroat Geothlypis aequinoctialis (Gmelin) is reported from Brazil. Isospora piacobrai n. sp. has oöcysts which are subspheroidal to ovoidal, measure 15.8 x 10.5 microm and have a smooth, bilayered wall c. 1.2 microm thick. A micropyle and oöcyst residuum are absent, but a polar granule is present. Its sporocysts are ovoidal and measure 15.8 x 10.5 microm. The Stieda body is knob-like and prominent, the substieda body large, trapezoidal and homogeneous, and the sporocyst residuum composed of granules of different sizes. Sporozoites are vermiform with one refractile body and a nucleus. This is the first description of an isosporoid coccidian infecting a bird of the family Parulidae.

Two new Isospora species from Brazilian tanager (Ramphocelus bresilius dorsalis) of South America.

Two new coccidian (Protozoa: Apicomplexa: Eimeriidae) species from the Brazilian tanager Ramphocelus bresilius dorsalis are reported in the current study. Isospora cadimi n. sp. oocysts are spheroidal to sub-spheroidal, 24.2 x 22.9 microm, with a smooth and bi-layered wall, approximately 1.1 microm. Micropyle, oocyst residuum, and polar granule are absent. Sporocysts are broadly ovoidal, 16.9 x 11.6 microm. Stieda and substieda bodies are present. Sporocyst residuum is present and sporozoites have refractile body and nucleus. Isospora navarroi n. sp. oocysts are spheroidal to sub-spheroidal, 21.4 x 20.6 microm, with a smooth and bi-layered wall, approximately 1.1 microm. Micropyle, oocyst residuum, and polar granule are absent. Sporocysts are ellipsoidal, 16.1 x 10.2 microm. Stieda and substieda bodies are present. Sporocyst residuum is present and sporozoites have a robust posterior refractile body.

New species of Eimeria Schneider, 1875 and Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) from the short-crested flycatcher Myiarchus ferox (Gmelin) (Passeriformes: Tyrannidae) in South America.

In the current study, two new coccidian species (Protozoa: Apicomplexa: Eimeriidae) obtained from short-crested flycatcher Myiarchus ferox (Gmelin) are reported from Brazil. Isospora feroxis n. sp. has oöcysts which are spheroidal to sub-spheroidal, 18.7 x 18.0 microm, with a smooth and bi-layered wall, c. 1.2 microm. The micropyle and oöcyst residuum are absent, but two polar granules are present. Its sporocysts are broadly ellipsoidal and 11.7 x 8.5 microm. Stieda and substieda bodies are present. A sporocyst residuum is present and the sporozoites have a refractile body and nucleus. Oöcysts of Eimeria sicki n. sp. are spheroidal to sub-spheroidal, 30.3 x 28.5 microm, with a smooth and bi-layered wall, c. 1.3 microm. The micropyle, oöcyst residuum and polar granule are absent. Its sporocysts are ellipsoidal, 18.4 x 10.0 microm. Stieda and substieda bodies are present. A sporocyst residuum is present and sporozoites have a refractile body and nucleus.

A new species of Amblyomma (Acari: Ixodidae) associated with monkeys and passerines of the Atlantic rainforest biome, Southeastern Brazil.

Recent studies have reported several larvae of an unidentified Amblyomma species on passerine birds in Atlantic rainforest fragments in southeastern Brazil. These larvae yielded a unique 16S rRNA haplotype designated as Amblyomma sp. haplotype Nazaré, which showed nucleotide identity levels of 91% to Amblyomma parkeri Fonseca & Aragão, 1952 and 88% to Amblyomma longirostre (Koch, 1844). Herein, we describe Amblyomma sp. haplotype Nazaré as a new species, Amblyomma romarioi n. sp. Martins, Luz & Labruna, through a formal description of the male and female adult stages. Amblyomma romarioi is morphologically and genetically most closely related to A. parkeri, A. longirostre and Amblyomma geayi Neumann, 1899. Among males, the rectangular basis capituli and rounded coxa I spurs separates A. romarioi from A. parkeri, A. longirostre, and A. geayi, which have basis capituli triangular or slightly hexagonal, and pointed coxa I spurs. Among females, the V-shaped genital aperture and coxa I rounded spurs of A. romarioi contrasts to the U-shaped genital aperture and coxa I pointed spurs in A. parkeri, A. longirostre, and A. geayi. Larvae of A. romarioi have been collected on 24 species of passerines. The few records of nymphs and adults were on the black-fronted titi monkey Callicebus nigrifrons (Spix, 1823). The current distribution of A. romarioi is restricted to the Brazilian Atlantic rainforest, southeastern Brazil, in areas with altitude between 363 and 1600 m, within the distribution of C. nigrifrons. We discuss ecological features of Amblyomma romarioi, comparatively to A. parkeri, A. longirostre and A. geayi. The present study increases the Brazilian tick fauna to 74 species.

First record of Ornithodoros faccinii (Acari: Argasidae) on toads of genus Rhinella (Anura: Bufonidae) in Brazil.

Although a group of soft ticks (Argasidae) associated with amphibians was recently discovered in Brazilian rainforests, parasitism by these ticks on cold-blooded animals remains less common than on mammal and bird species. In this study, we identified ticks that were collected from toads that had been caught in December 2016 and January 2017, at Itinguçú waterfall (22°54'05" S; 43°53'30" W) in the municipality of Itaguaí, state of Rio de Janeiro. Tick specimens were identified using a morphological and molecular approach. In total, twelve larvae of Ornithodoros ticks were collected from three individuals of Rhinella ornata and were identified as Ornithodoros faccinii. Our results include a longer 16S rRNA mitochondrial sequence for O. faccinii that supports its phylogenetic relatedness to Ornithodoros saraivai, and we report this tick species parasitizing Rhinella toads for the first time in Brazil.

Infection of Amblyomma ovale with Rickettsia species Atlantic rainforest in Serra do Mar, São Paulo State, Brazil.

Rickettsia rickettsii and Rickettsia sp. strain Atlantic rainforest, that is considered to represent a genetic variant of Rickettsia parkeri, are confirmed as being capable of infecting humans in Brazil. This study reports the detection and characterization, by PCR and nucleotide sequencing, of Rickettsia sp. strain Atlantic rain forest in Amblyomma ovale parasitizing a human, in ticks infesting dogs and in free-living ticks collected from the environment where the human infestation was recorded. The data contribute to our knowledge of infection rates in A. ovale with Rickettsia sp. strain Atlantic rainforest and identified an additional location in the state of São Paulo populated with ticks infected with this emerging pathogen.

Oocyst shedding by green-winged-saltator (Saltator similis) in the diagnostic of coccidiosis and Isospora similisi n. sp. (Apicomplexa: Eimeriidae).

Diurnal periodicity is a phenomenon that has been observed in coccidian of Isospora parasites of passerines, which have been eliminated great number of oocysts at dusk. The objective of this study was to evaluate the occurrence of periodicity of oocysts presence in the green-winged-saltator Saltator similis, and its use in the diagnosis of coccidiosis in wild birds in captivity. A total of 220 fecal samples were collected from birds, apprehended from illegal trading and kept in quarantine in CETAS∕IBAMA, in the morning and late afternoon, from May to November 2010. It was observed that 1.82% of the samples collected in the morning were positive, while 31.36% of samples were positive in the late afternoon. In addition, the number of oocysts shed was greater in the afternoon. Therefore, it was concluded that the sampling in the late afternoon provided greater reliability for the diagnosis of coccidiosis in green-winged-saltators. Moreover, in this study a new isosporoid coccidian parasite from the green-winged-saltator S. similis was observed and is herein described. Isospora similisi n. sp. oocysts are spheroidal to sub-spheroidal, 27.5 × 25.9 µm, with a smooth and bi-layered wall, ∼1.2 mm. Micropyle and oocyst residuum are absent, but splinter-like or comma-like granules are present. Sporocysts are ellipsoidal or slightly ovoidal, 17.4 × 12.2 mm. A stieda body and substieda body are present. The sporocyst residuum is composed of granules of different sizes. Sporozoites are vermiform with a single refractile body and a nucleus. This is the fourth description of an isosporoid coccidium infecting S. similis and the sixth description from Cardinalidae.

First record of Ornithodoros faccinii (Acari: Argasidae) on toads of genus Rhinella (Anura: Bufonidae) in Brazil / Primeiro registro de Ornithodoros faccinii (Acari: Argasidae) em sapos do gênero Rhinella no Brasil

Abstract Although a group of soft ticks (Argasidae) associated with amphibians was recently discovered in Brazilian rainforests, parasitism by these ticks on cold-blooded animals remains less common than on mammal and bird species. In this study, we identified ticks that were collected from toads that had been caught in December 2016 and January 2017, at Itinguçú waterfall (22°54'05" S; 43°53'30" W) in the municipality of Itaguaí, state of Rio de Janeiro. Tick specimens were identified using a morphological and molecular approach. In total, twelve larvae of Ornithodoros ticks were collected from three individuals of Rhinella ornata and were identified as Ornithodoros faccinii. Our results include a longer 16S rRNA mitochondrial sequence for O. faccinii that supports its phylogenetic relatedness to Ornithodoros saraivai, and we report this tick species parasitizing Rhinella toads for the first time in Brazil.

Resumo Embora um grupo de carrapatos moles (Argasidae) associado a anfíbios tenha sido recentemente descoberto nas florestas brasileiras, o parasitismo por esses carrapatos em animais de sangue frio permanece menos comum do que nas espécies de mamíferos e aves. Neste estudo, identificamos carrapatos que foram coletados de sapos capturados em dezembro de 2016 e janeiro de 2017, na cachoeira de Itinguçú (22°54'05" S; 43°53'30" W) no município de Itaguaí, estado do Rio de Janeiro. Os espécimes de carrapatos foram identificados usando uma abordagem morfológica e molecular. No total, doze larvas de carrapatos Ornithodoros foram coletadas de três indivíduos de Rhinella ornata e foram identificadas como Ornithodoros faccinii. Nossos resultados incluem uma maior seqüência mitocondrial 16S rRNA para O. faccinii que suporta sua relação filogenética com Ornithodoros saraivai e relatamos esta espécie de carrapato parasitando sapos Rhinella pela primeira vez no Brasil.

Isospora coerebae n. sp. (Apicomplexa: Eimeriidae) from the bananaquit Coereba flaveola (Passeriformes: Coerebidae) in South America.

The present study describes a new isosporoid coccidian parasite from the bananaquit Coereba flaveola, in Brazil. This new species is similar to I. cagasebi, but it can be distinguished by the size and shape of Stieda and susbstieda bodies. Isospora coerebae n. sp. oocysts are spheroidal to sub-spheroidal, 24.8 × 23.3 µm, with a smooth and bi-layered wall, ~1.2 µm. Micropyle, oocyst residuum and polar granule are absent. Sporocysts are elongate ovoidal, 17.9 × 10.9 µm. Stieda and substieda bodies are present. Sporocyst residuum is present and sporozoites have a posterior refractile body.

Three new species of Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) from the buffy-fronted seedeater Sporophila frontalis Verreaux (Passeriformes: Emberizidae) in South America.

Three new coccidian (Apicomplexa: Eimeriidae) species from the buffy-fronted seedeater Sporophila frontalis Verraux in Brazil are reported in the current study. Isospora frontalis n. sp. oöcysts are spherical to sub-spherical, 27.9 x 26.9 mum, with a smooth, bi-layered wall c.1.4 microm thick. A micropyle and an oöcyst residuum are absent, but polar granules are present. Sporocysts are elongate ellipsoidal, 19.6 x 11.1 microm. The Stieda body is knob-like and the substieda body is slight. The sporocyst residuum is composed of scattered granules, and the sporozoites are vermiform with a refractile body and a nucleus. Isospora teresopoliensis n. sp. oöcysts are spherical to sub-spherical, 25.7 x 24.3 microm, with a smooth, bi-layered wall 1.3 microm thick. A micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ovoid, 18.8 x 11.2 microm. The Stieda body is nipple-shaped and the substieda body is large and prominent. The sporocyst residuum is composed of scattered granules, and the sporozoites are large and elongate, with a refractile body and nucleus. Oöcysts of Isospora chanchaoi n. sp. are spherical to sub-spherical or ovoid, 24.2 x 22.0 microm, with a smooth, bi-layered wall c.1.2 microm thick. Both a micropyle and an oöcyst residuum are absent, but one or two polar granules are present. The sporocysts are ellipsoidal, 16.1 x 10.3 microm. The Stieda body is nipple-shaped and the substieda body is small but prominent. The sporocyst residuum forms a compact mass of granules, and the sporozoites are large and elongate, with a refractile body and a nucleus.

Eimeria divinolimai sp. n. (Apicomplexa: Eimeriidae) in the rufous casiornis Casiornis rufus Vieillot, 1816 (Passeriformes: Tyrannidae) in Brazil.

Eimeria divinolimai sp. n. from the rufous casiornis, Casiornis rufus (Passeriformes: Tyrannidae) was described in Brazil. Oocysts are subspherical 17.84 +/- 1.52 by 15.90 +/- 0.99microm (15.61-20.00 x 14.15-17.80). Shape-index (length/ width) of 1.12 +/- 0.05 (1.01-1.20). Wall smooth and bilayered, being yellowish outer and darker inner, 2.13 +/- 0.16 microm (2.00-2.38) thick. Micropyle and residuum are absent, but one subspherical polar granule is present. Sporocysts are ovoid ranging from 14.98 +/- 0.85 by 7.50 +/- 0.44 microm (13.81-1619 x 6.76-8.09), with smooth, thin and single-layered wall. Stieda body prominent, without substiedal body and with residuum granulated. Sporozoites with refractile body at one end.

First report of Tyzzeria parvula (Kotlán, 1933) Klimes, 1963 (Apicomplexa: Eimeriidae), in swan goose Anser cygnoides L., 1758, in Brazil.

This is the first description of Tyzzeria parvula from the swan goose (Anser cygnoides) in the State of Rio de Janeiro, Brazil. Fecal samples were collected from 15 swan geese, but only four of them shed oocysts in feces. After sporulation, the oocysts were spherical to sub-spherical, and measured 12.4 +/- 1.0 (11-15) x 10.4 +/- 0.8 mum (8-12). They presented a shape index of 1.2 (1.0-1.4), with a bi-layered wall that was 0.7 +/- 0.1 mum (0.6-0.8) thick. The outer side was smooth and colorless, while the inner side was pale greenish. Micropyles and polar granules were absent. The residuum sometimes formed a mass of spherules and granules and, at other times, was dispersed. Sporozoites had one rounded end and the other end was fine and slightly curved.

Contribution on the study of Isospora hemidactyli Carini, 1936 and a report of an adeleid pseudoparasite of the house gecko Hemidactylus mabouia, from the Rio de Janeiro Metropolitan Region, Brazil.

A description of the coccidium Isospora hemidactyli from the house gecko Hemidactylus mabouia, a very common at dwellings in Rio de Janeiro Metropolitan Region, was made in this study. Histograms and linear regression were made for this species and determined the homogeneity of these oocysts despite of large range. Besides it, polysporocystid oocysts also were recovered from feces of the H. mabouia house gecko and they were similar to those described previously as the genus Adelina. This species should be parasitizing an invertebrate ingested by house gecko, and for that reason, is a pseudoparasite. Oocysts of I. hemidactyli were subspherical to ellipsoidal, 24.4 x 22.3 microm, with single-layered wall and one polar granule. Sporocysts were subspherical to ellipsoidal, 11.8 x 9.8 microm with Stieda and substieda bodies, residuum and sporozoites with refractile body. Oocysts of the pseudoparasite Adelina sp. were ellipsoidal, 36.3 x 30.9 microm, with bi-layered wall and without micropyle, residuum and polar granule. Eight to 15 sporocysts were presents and were subspherical to broadly ellipsoidal, 12.4 x 11.2 microm. Stieda and substieda bodies were absent. Sporozoites present refractile bodies at both ends.