Molecular diversity and polyparasitism of avian trypanosomes in the Brazilian Atlantic Rainforest.

The current study proposes to investigate the diversity and phylogeny of trypanosomes parasitizing wild birds from the Brazilian Atlantic Forest. Cytological examination was carried out by light microscopy of blood smears and positive birds were selected for amplification of the 18S rDNA sequence through PCR. The resulting amplicons were subjected to purification, cloning, and sequencing analysis. Phylogenetic reconstruction was conducted, including all avian trypanosomes representative's lineages. A total of ten bird samples from species of Turdus flavipes (N=1/12), T. albicollis (N=1/8), Tachyphonus coronatus (N=6/121), Thamnophilus caerulescens (N=1/22) and Synallaxis spixi (N=1/8) were positive for Trypanosoma spp. In the six specimens of T. coronatus, five distinct lineages of Trypanosoma spp. 18S-rRNA were observed in ninety sequences obtained, and using the strategy of cloning independent PCR, it was possible to observe that two of them were related to T. avium (JB01/JB02), and three were closed related to T. bennetti (JB03/ JB04/JB05). Addionaly, all fifteen sequences obtained from T. caerulescens/ S. spixi/T. flavipes/T. albicollis were identical. The present research is the first study to access molecular diversity and polyparasitism by avian trypanosomes in Brazil. The current research exhibits the wide genetic variability in avian trypanosomes and its non-specific relationship with its avian hosts.

High infestation and phylogenetic position of Epistylis sp. (Ciliophora, Peritrichia) on Aegla serrana Buckup & Rossi (Crustacea, Anomura) from southern Brazil.

High infestations of epibiont ciliates on vertebrates or invertebrates are normally related to aquaculture tanks or similar environments, and the importance of this relationship in natural habitats is often disregarded. Here, we describe the first record of high infestation of ciliates on Aegla serrana in South America and conduct a brief morphological and phylogenetic characterization of these ciliates. Our findings confirm that cases of high infestation of ciliates on metazoans can indeed occur in natural environments.

Insights into the systematics of the family Ophryoscolecidae (Ciliophora, Entodiniomorphida).

The family Ophryoscolecidae currently comprises 225 species of trichostomatid ciliates, subdivided into three subfamilies (Entodiniinae, Diplodiniinae, and Ophryoscolecinae). The last taxonomic review of the family was performed 55 years ago, but recent morphological and molecular studies indicate the need for a profound review of the systematics of this taxon, since its current taxonomy is insufficient to organize the diversity of the group. Here, we briefly review the systematics of the family Ophryoscolecidae based on information recovered from the literature and new morphological and molecular data. We add four new 18S rDNA sequences of ophryoscolecids to molecular databases, which contributed to improving the comprehension of intrafamily relationships within this group. Finally, we discuss some systematic problems and suggest approaches to resolve such inconsistencies in the future.

New Finds on the Systematics of Cycloposthiid Ciliates (Ciliophora: Entodiniomorphida: Cycloposthiidae) Based on New 18S-rDNA Sequences from a Brazilian Capybara.

The family Cycloposthiidae (Entodiniomorphida) comprises ciliated protists that are symbionts of the gastrointestinal tract of several herbivore mammals, such as rodents, elephants, equids, primates, hippopotamus, marsupials, rhinoceros and tapir, where they contribute to the digestion of their host's plant-based diet. Despite the significance of these ciliates to the evolution of their hosts, many characters used in the taxonomy of the group are homoplastic and most of the valid species do not have molecular data available. For these reasons the systematics of this family is poorly understood. Here, we sequenced the 18S-rDNA of ten cycloposthiids, including nine Cycloposthium spp. and Monoposthium cynodontum, all of them isolated from the cecum of a Brazilian capybara. Our phylogenetic analyses indicate that the family Cycloposthiidae might be polyphyletic, while M. cynodontum and Cycloposthium spp. constitute a single monophyletic group. Given the great morphological and molecular similarities between members of M. cynodontum and Cycloposthium ciliates, it is possible that this species, although it has been described in the genus Monoposthium, is actually a Cycloposthium ciliate.

Rediscovery of Plasmodium (Huffia) huffi (Apicomplexa, Haemosporida): a lost lineage from toucans.

Literature data on toucans haemosporidians are scarce and all reports come from investigations in Brazil. Muniz et al. (Rev Bras Malariol 3: 339-356, Muniz et al., Rev Bras Malariol 3:339-356, 1951) and Muniz and Soares (Rev Bras Malar 611-617, Muniz J, Soares R de RL (1954) Nota sôbre um parasita do gênero Plasmodium encontrado no Ramphastos toco Müller, 1776, "Tucano-Açu", e diferente do Plasmodium huffi: Plasmodium pinottii n. sp. Rev Bras Malar 611 - 617.) described two Plasmodium species, P. huffi and P. pinottii, in Ramphastos toco. Later, Manwell and Sessler (J Protozol 18: 570-574, Manwell and Sessler, Malaria Parasites of Toucans J Protozol 18:570-574, 1971) established a new subspecies, P. nucleophilum toucani. In the last review on avian haemosporidians, Valkiunas (Valkiunas, Avian malaria parasites and other haemosporidia, CRC Press, New York, 2005) highlighted that P. huffi was insufficiently characterized, considering it a lost lineage. Also, the original description of P. huffi was considered insufficiently clear, due to a possible co-infection of the toucan hosts with a Novyella-like species. Here, we redescribed the species Plasmodium (Huffia) huffi based on morphological and molecular data, which were found in two toucan species, Ramphastos toco and Pteroglossus aracari from Brazil. Morphological features of the specimens are markedly the same as the original description. In R. toco, we observe two individuals infected, one infected only with P. huffi and one co-infected with P. huffi and the Novyella-like species, as observed in the original description. Also, we observe one R. toco infected only with the Novyella-like species, identified by morphological and molecular data as Plasmodium (Novyella) nucleophilum nucleophilum. In this way, it was possible to redescribe Plasmodium huffi in detail, without the doubt characters observed in the original description. Moreover, by applying species delimitation algorithms to a large Plasmodium phylogeny, we were able to identify new possible hosts for P. huffi and extend its geographic distribution to include North America.

New morphological and molecular data for Haemoproteus (H.) paramultipigmentatus in the Atlantic Forest of Brazil.

Parasites of the genus Haemoproteus have been reported in almost all avian clades and zoogeographic regions, except Antarctica. However, despite the large number of reports worldwide, they are poorly studied in the Neotropical region, which includes the Atlantic Forest, a biodiversity hotspot with more than 890 bird species, of which 24% are endemic. Haemoproteus (Haemoproteus) paramultipigmentatus was described by morphological and molecular analyses in 2013 infecting Columbiform birds in Mexico. However, since the original description this parasite has not been studied in detail. Here, we investigate the prevalence of Haemoproteus spp. in Brazilian Columbiformes and conducted a taxonomic integrative study of the species Haemoproteus (Haemoproteus) paramultipigmentatus, including new morphological and molecular data from a Brazilian population. Moreover, we provide discussions about the geographic distribution and phylogenetic relationships between different lineages of this parasite. Our findings demonstrated a high prevalence of Haemoproteus spp. infection in Brazilian Columbiformes, which is in accordance with previous studies. Morphological characterization of H. paramultipigmentatus revealed minor differences from the original description. Through molecular and phylogenetic analyses we identified a new lineage of H. paramultipigmentatus that was added to the genetic databases. Our findings also suggest a new geographical distribution for this hemoparasite, including South American countries, and raise discussions about its current distribution.

Ciliates as bioindicators of water quality: A case study in the neotropical region and evidence of phylogenetic signals (18S-rDNA).

The aim of our study was to evaluate the water quality of an urban stream in southeastern Brazil by analyzing epibenthic ciliates, and to investigate the existence of phylogenetic signal for saprobity in ciliates. However, before conducting this type of phylogenetic study, it is necessary to evaluate if the saprobic classification used frequently in the northern Hemisphere is suitable for neotropical ecosystems. Sediment samples were collected from five sampling stations: two in rural areas and three in urban areas. During the one-year study, with monthly collections, 39 ciliates species were found, of which 32 are included in the saprobic system. Physical, chemical and biological parameters of water confirm the spatial heterogeneity of the sampling stations, with a clear influence of organic pollution on the composition and structure of ciliates taxocenosis. The saprobic index and the saprobic valence index were used to evaluate the water quality of the sampling stations and demonstrated clear heterogeneity between the stations and high degree of pollution of the urban area. These sampling stations were dominated by ciliates indicators of polysaprobric environments. Since we were able to successfully use the saprobic index in a limnic ecosystem in Brazil, we applied the phylogenetic signal validation as a tool for saprobity prediction of the limnic ciliate species not yet analyzed. A phylogenetic tree containing only 18S-rDNA nominal sequences of freshwater ciliates was estimated and used to explore the existence of the phylogenetic signal, which showed that the sensitivity/tolerance of ciliates to organic pollution reflected evolutionary divergence. The results confirm the existence of phylogenetic signal for the saprobrity in Ciliophora. Also, our results suggest that evolutionary analysis is a potential method to predict lineages of ciliates not yet classified for saprobity.

Haemoproteus syrnii (Haemosporida: Haemoproteidae) in owls from Brazil: morphological and molecular characterization, potential cryptic species, and exo-erythrocytic stages.

Haemoproteus syrnii is a haemosporidian parasite found in owls. Although morphological and molecular data on the species is available, its exo-erythrocytic development was never researched. In this study, we provide the morphological, morphometric, and molecular characterization of H. syrnii populations found in owl species from Minas Gerais, southeast Brazil. We also characterized the coalescent species delimitation based on the molecular and histopathology data. Samples from 54 owls from six different species were analyzed, generating 11 sequences of the cyt b gene, from which six were new sequences. The overall prevalence of infection was high (72.22%). The H. syrnii sequences were grouped into two well-supported independent clades, which included other Haemoproteus (Parahaemoproteus) species. This was supported by both the coalescent species delimitation analysis and by the genetic divergence between lineages of these distinct clades. There were small morphological and morphometric differences within the population presented in this study. However, when compared with other studies, the molecular analysis demonstrated considerable intraspecific variation and suggests potential cryptic species. The histopathological analysis revealed, for the first time, that lungs and skeletal muscle are exo-erythrocytic stage location of H. syrnii, and that the parasite is linked to the histopathological changes found in owls. This study brings new data from Haemoproteus species biology and host infection, and improves host-parasite relationship understanding under an owl conservation perspective.

Molecular diversity and coalescent species delimitation of avian haemosporidian parasites in an endemic bird species of South America.

Haemoproteus spp. and Plasmodium spp. are blood parasites that occur in birds worldwide. Identifying the species within this group is complex, especially in wild birds that present low parasitemia when captured, making morphological identification very difficult. Thus, the use of alternative tools to identify species may be useful in the elucidation of the distribution of parasites that circulate in bird populations. The objectives of this study were to determine the prevalence and parasitemia of the genera Plasmodium and Haemoproteus in Tachyphonus coronatus in the Atlantic Forest, Brazil, and to evaluate the molecular diversity, geographic distribution, and specificity of these parasites based on coalescent species delimitation methods. Microscopic analysis, PCR, cyt b gene sequencing, phylogenetic analysis and coalescent species delimitation using single-locus algorithms were performed (Poisson tree process (PTP) and multi-rate Poisson tree process (MPTP) methods). The analyses were performed in 117 avian host individuals. The prevalence was 55.5% for Plasmodium and 1.7% for Haemoproteus, with a mean parasitemia of 0.06%. Twenty-five Plasmodium and two Haemoproteus lineages were recovered. The MPTP method recovered seven different evolutionarily significant units (ESUs) of Plasmodium and one of Haemoproteus, whereas PTP presented fourteen ESUs of Plasmodium and one of Haemoproteus. The MPTP was more consistent with current taxonomy, while PTP overestimated the number of lineages. These ESUs are widely distributed and have already been found in 22 orders of birds that, all together, inhabit every continent, except Antarctica. The computational methods of species delimitation proved to be effective in cases where the classification of Haemosporida based just on morphology is insufficient.

Morphological and molecular characterization of Haemoproteus coatneyi and Haemoproteus erythrogravidus (Haemosporida: Haemoproteidae) in Passeriformes in Brazil's Atlantic Forest.

Haemoproteus spp. are protozoan parasites found in birds around the world. These parasites are identified through the morphology of gametocytes, phylogenetic analysis based on the mitochondrial cytb gene, and the parasite's geographic distribution. The absence of erythrocytic merogony, high intraspecific genetic variation and low parasitemia in wild birds makes it essential to use integrative approaches that assist in the identification of these parasites. Thus, microscopic and molecular analyses, combined with spatial distribution, were carried out to verify the presence of Haemoproteus spp. in wild birds in Brazil. Light microscopy revealed one Tangara sayaca bird was parasitized by Haemoproteus coatneyi and, two specimens of Zonotrichia capensis presented Haemoproteus erythrogravidus. The morphology of the gametocytes of these two parasitic species showed high similarity. The molecular analysis revealed the presence of one lineage of H. coatneyi and two lineages of H. erythrogravidus, one of which is considered a new lineage. These lineages were grouped phylogenetically in separate clades, with low genetic divergence, and the H. erythrogravidus lineage emerged as an internal group of the lineages of H. coatneyi. The geographic distribution demonstrated that the two species occur in the American continent. This is the first report of H. erythrogravidus in Brazil.

Helmet-shaped Body of Entodiniomorphid Ciliates (Ciliophora, Entodiniomorphida), a Synapomorphy or a Homoplasy?

Triadinium was created to include Triadinium caudatum. Further, four other species were included, T. minimum, T. galea, T. elongatum, and T. magnum, all sharing a characteristic helmet-shaped body. Wolska and Grain argued that the inclusion of T. minimum and T. galea into Triadinium was done based on superficial morphological aspects, and established two new genera to accommodate these species: Circodinium and Gassovskiella. Although the phylogenetic relationships within Entodiniomorphida have been investigated by multiple authors, none of them discussed the evolutionary relationship of helmet-shaped entodiniomorphids. We performed molecular phylogenetics and revisited old literature digging for morphological data to explain our results. According to our analyses, the helmet-shaped body is homoplastic and may have evolved from at least three different entodiniomorphid ancestors. Circodinium minimum is phylogenetically related to members of Blepharocorythidae, T. caudatum emerged within Spirodiniidae and G. galea within Polydiniellidae. This phylogenetic hypothesis is partially supported by information on infraciliature and ultrastructure of C. minimum and T. caudatum. However, such morphological information is not available for polydiniellids. In order to shed some light into the evolution of the helmet-shaped ciliates, future works should focus to collect information on the infraciliature and the ultrastructure of Polydiniella mysorea and of other Triadinium species.

Intestinal ciliates (Alveolata, Ciliophora) in Brazilian domestic horses (Equus caballus L.) and a review on the ciliate communities associated with horses around the world.

The aims of this study were to identify and quantify the gastrointestinal ciliates from Brazilian domestic horses, to compare the data obtained with previous studies and to review data on ciliate communities associated with horses around the world. We used eight domestic horses, kept under extensive management. Fecal samples were obtained immediately after defecation of animals and fixed in 18.5% formaldehyde. We identify 24 species of endosymbiotic ciliates, distributed in two subclasses, two orders, five families and 13 genera. The ciliate composition was similar among the animals analyzed. The family Blepharocorythidae presented the higher prevalence and the family Buetschliidae was observed in greater abundance and density. Among the species identified, Cycloposthium dentiferum was observed for the first time in Central and South America and Bundleia elongata, B. inflata, B. piriformis, B. vorax and Cycloposthium psicauda, were observed for the first time in the American continent. In addition, the species Buissonella tapiri, which was originally discovered in fecal contents of the American tapir (Tapirus terrestris) in Brazil was observed for the first time associated to Equus caballus. The review on the ciliate communities associated with horses around the world demonstrated that the equine ciliates are a very diversified group with representatives in two subclasses, Suctoria and Trichostomatia, totaling 78 species distributed in three orders and six families, having been registered in different equine hosts (Equus asinus, Equus caballus and Equus quagga) and geographic locations (USA, China, Russia, Taiwan, Mexico, Cyprus, Turkey, Kyrgyzstan and Brazil).

Morphological and morphometric characterization of trypanosomes in Leptodactylus lineatus and Osteocephalus sp. (Anura) from Brazilian Midwest / Caracterização morfológica e morfométrica dos tripanossomas de anuros do Centro-Oeste brasileiro

Este estudo teve por objetivo avaliar o parasitismo causado por tripanossomas em anuros naturalmente infectados do Centro-Oeste brasileiro e realizar a caracterização morfológica e morfométrica dos tripanossomas. Os anuros foram capturados às margens do Rio Guaporé, na transição entre os biomas Cerrado e Floresta Amazônica. Amostras de sangue de 16 espécies de anuros foram obtidas por punção cardíaca e analisadas para a presença de hemoparasitos. Tripanossomas foram encontrados infectando duas espécies de anuros, Leptodactylus lineatus (Leptodactylidae) e Osteocephalus sp. (Hylidae). Foi observada alta prevalência dos tripanossomas (100%; N = 3 hospedeiros) em L. lineatus, com intensidade da infecção calculada em 9.9 x 102 parasitos/ml. Em Osteocephalus sp., a prevalência foi comparativamente menor (36%; N = 4 hospedeiros infectados de 11 analisados), no entanto, a intensidade de infecção foi maior 2.16 x 103 parasitos/ml. Variações morfológicas relacionadas ao comprimento e largura do corpo foram observadas entre as diferentes espécies de hospedeiros e também em indivíduos da mesma espécie. Os tripanossomas polimórficos foram separados em morfotipos classificados como corpo alongado ou curto. Os tripanossomas que infectam L. lineatus possuem o corpo alongado que pode ser delgado ou largo e aqueles que infectam Osteocephalus sp. apresentam o corpo curto com a extremidade posterior alargada. A análise parasitológica e a caracterização morfológica apresentada neste estudo contribuem para o conhecimento da diversidade dos tripanossomas de anuros neotropicais.

The objective of this study was to evaluate parasitism caused by trypanosomes in naturally infected anurans from Brazilian Midwest and characterize trypanosomes by morphology and morphometry. Anurans were captured from margins of the Guaporé River in transition areas between Cerrado and Rain Forest. Blood samples of 16 anurans species were obtained by cardiac puncture and analyzed for the presence of hemoparasites. Trypanosomes were found infecting two anuran species, Leptodactylus lineatus (Leptodactylidae) and Osteocephalus sp. (Hylidae). It was observed high prevalence of trypanosomes (100%; N = 3) in L. lineatus, with intensity of infection of 9.9 × 102 parasites/ml. In Osteocephalus sp. the prevalence of trypanosomes was comparatively lower (36%; N = 4 from 11), however, the intensity of infection was higher 2.16 × 103 parasites/ml. Morphological variation related to body length and width of bloodstream trypomastigotes were observed in different host species and between individual hosts of the same species. Polymorphic trypanosomes were separated into morphotypes and classified as elongated or short trypanosomes. Trypanosomes infecting L. lineatus are elongated with slender or broad body and trypanosomes infecting Osteocephalus sp. presented a short body with broad posterior extremity. The parasitological analyses and morphological characterizations presented in this study contribute to the knowledge of diversity of trypanosomes from neotropical anurans.

Polymorphism evidence in Plasmodium (Haemamoeba) lutzi Lucena, 1939 (Apicomplexa, Haemosporida) isolated from Brazilian wild birds.

Plasmodium parasites can infect great variety of bird species around the world inflicting the so called avian malaria, an illness that could be fatal in some cases and consequently, should be monitored and widely included into conservation programs. The aim of this study was to characterize two lineages of Plasmodium (Haemamoeba) lutzi found in some birds in the Atlantic Forest of Minas Gerais - Brazil, that were morphologically identified after blood smears analyses under light microscopy and molecularly by sequencing the mitochondrial cytochrome b gene (cyt b). Besides these two lineages could be clearly morphologically identified as P.(H.) lutzi, some variations in comparison with its original description were noticed: absence of meronts and gametocytes (early and fully grown) in polychromatic erythrocytes, the larger size of pigment granules in meronts and gametocytes, and the presence of small vacuoles between pigment accumulation in fully grow macrogametocytes. Moreover, a certain degree of genetic intraspecific diversity was also observed across the lineages of P. (H.) lutzi, indicating the existence of polymorphisms within this taxon, which is uncommon in Haemosporida. These results allow discussion about species boundaries within avian hemosporidians and highlight the importance of multidisciplinary approaches for a more efficient species identification and characterization.

Trichostomatid Ciliates (Alveolata, Ciliophora, Trichostomatia) Systematics and Diversity: Past, Present, and Future.

The gastrointestinal tracts of most herbivorous mammals are colonized by symbiotic ciliates of the subclass Trichostomatia, which form a well-supported monophyletic group, currently composed by ∼1,000 species, 129 genera, and 21 families, distributed into three orders, Entodiniomorphida, Macropodiniida, and Vestibuliferida. In recent years, trichostomatid ciliates have been playing a part in many relevant functional studies, such as those focusing in host feeding efficiency optimization and those investigating their role in the gastrointestinal methanogenesis, as many trichostomatids are known to establish endosymbiotic associations with methanogenic Archaea. However, the systematics of trichostomatids presents many inconsistencies. Here, we stress the importance of more taxonomic works, to improve classification schemes of this group of organisms, preparing the ground to proper development of such relevant applied works. We will present a historical review of the systematics of the subclass Trichostomatia highlighting taxonomic problems and inconsistencies. Further on, we will discuss possible solutions to these issues and propose future directions to leverage our comprehension about taxonomy and evolution of these symbiotic microeukaryotes.

Rhabdostylid Ciliates (Ciliophora, Peritrichia, Epistylididae) as Epibionts on Chironomid Larvae: Evidence of High Specificity and Association with Organic Pollution.

Although epibiont ciliates make up a significant part of the biomass in aquatic ecosystems and may cause perceptible alterations in the population dynamics of their hosts, studies on the extrinsic and intrinsic factors that control the abundance of these microorganisms are scarce in literature. In the present study, we investigated the colonization site and intensity of rhabdostylid epibiont upon chironomid larvae and assessed the influence of organic pollution and chironomid communities on the prevalence and abundance of epibiont ciliates at five sampling stations (two in rural areas and three in urban areas) along a neotropical urban stream over a period of 12 months. Among the 24 genera of chironomids found and the 31,976 larvae analyzed, 96.12% belonged to the Chironomus genus, of which 16.95% (5212) were colonized by Rhabdostyla aff. chironomi. The infestation intensity varied from one to 67 individuals per host with an average intensity of 4.86 (± 33.45). Ciliates were only found colonizing the chironomids' ventral tubules. The high number of chironomid larvae, high host- and site-specificity, low infestation intensity, and absence of apparent structural damage to hosts evidence an intimate relationship between epibiont and basibiont as well as a possibly long coevolutionary history. Both prevalence and abundance of epibiont ciliates were correlated to the pollution rate of the studied stream. There was an alteration in the composition and structure of the chironomid community along the sampling stations. Numeric dominance of Chironomus-tolerant chironomids and its direct correlation to infestation prevalence and to ciliates abundance highlights the predilection of both peritrich ciliates and Chironomus larvae for organically-enriched environments.

Morphology and description of infraciliary bands pattern in four Metadinium Awerinzew Mutafowa, 1914 species (Ciliophora, Entodiniomorphida, Ophryoscolecidae) with taxonomic notes on the genus.

The genus Metadinium includes ophryoscolecid ciliates, with two retractile ciliary zones in the anterior body portion, two broad skeletal plates, a lobed macronucleus and two contractile vacuoles. Species belonging to this genus were recorded in several geographical locations and in different host species. However, they were mostly described based on a single ciliatological technique, and until now, there is only one description of the infraciliary pattern in Metadinium ciliates (Metadinium medium). The present study performs the morphological and infraciliature characterization of four species belonging to Metadinium: M. esalqum, M. minorum, M. rotundatum and M. ypsilon; and presents taxonomic notes on the genus. The oral infraciliature in these four Metadinium species corresponds to the Diplodinium-type, pattern shared with genera Diplodinium, Diploplastron, Eudiplodinium, Eremoplastron and Polyplastron. A brief review conducted highlights the difficulty of delimiting Metadinium species due to the morphological similarity among them.

Diversity and Distribution of Peritrich Ciliates on the Snail Physa acuta Draparnaud, 1805 (Gastropoda: Physidae) in a Eutrophic Lotic System.

Bianca Sartini, Roberto Marchesini, Sthefane D ´ávila, Marta D'Agosto, and Roberto Júnio Pedroso Dias (2018) Freshwater gastropods represent good models for the investigation of epibiotic relationships because their shells act as hard substrates, offering a range of microhabitats that peritrich ciliates can occupy. In the present study we analyzed the community composition and structure of peritrich epibionts on the basibiont freshwater gastropod Physa acuta. We also investigated the spatial distribution of these ciliates on the shells of the basibionts, assuming the premise that the shell is a topologically complex substrate. Among the 140 analyzed snails, 60.7% were colonized by peritrichs. We observed seven peritrich species: Epistylis plicatilis and Epistylis sp. (Epistylididae); Opercularia articulata (Operculariidae); Carchesium polypinum, Vorticella campanula and Vorticella sp. (Vorticellidae) and Thuricola kellicottiana (Vaginicollidae). We observed a high prevalence of epibiosis (> 60%) when all species of ciliates were considered conjunctly. However, the prevalence was low (1-58%) when each species was considered separately, reflecting their aggregate distribution pattern. The most prevalent species were Epistylis sp. (58.60%), Vorticella sp. (14.30%) and O. articulata (13.60%). Although the epibionts were distributed through the shell's entire extension, we observed greater values of abundance, density, diversity and dominance on the dorsal surface. Only Epistylis sp. was widely distributed on the ventral surface. We also observed that the peritrichs predominantly occupied the areas of the shell delimited by the sutures. We interpreted these distribution patterns considering that the peritrichs select their attachment sites under pressures related to basibiont behavior, physical forces that may dislocate them and protective characteristics of the shell's areas.

Molecular and Morphological Characterization of a Brazilian Lineage of Plasmodium ( Novyella) Unalis in Turdus Spp. (Passeriformes) of the Atlantic Forest, with Remarks on New Hosts and High Genetic Variation.

Plasmodium spp. are haemosporidian protozoans that alternate their live cycles between bloodsucking Culicidae dipterans and vertebrate hosts (mammals, reptiles, and birds). In birds, these parasites are the causative agents of the so-called avian malaria, a disease associated with considerable declines and extinctions in the avifauna in different geographical regions. In this work, we applied a multidisciplinary approach, light microscopy and cytochrome oxidase b (cyt b) gene sequence analysis, for characterization of Plasmodium spp. found in association with wild birds of the genus Turdus, collected in Atlantic forest fragments of southeastern Brazil. From the total 90 analyzed birds, 58 (47 Turdus rufiventris, 9 Turdus leucomelas, 1 Turdus albicollis, and 1 Turdus flavipes) were positively infected with Plasmodium unalis, a haemosporidian that was previously detected in Turdus fuscater in Colombia and in penguins in Brazil, but has never been found in association with these Turdus species of this present work. Moreover, all 7 new sequences of P. unalis cyt b gene clustered into a monophyletic clade with previously characterized P. unalis sequences with a mean genetic divergence of 1.6% and with a maximum divergence of 3.1%, indicating for a high degree of intraspecific polymorphism within this parasitic species. Together, our data highlight the existence a high degree of intraspecific variation within P. unalis and highlight the importance of integrative taxonomy to an accurate identification and characterization of avian haemosporidian parasites.

Description of Diploplastron dehorityi sp. nov. (Entodiniomorphida, Ophryoscolecidae), a new rumen ciliate from Brazilian sheep (Ovis aries).

To date the genus Diploplastron comprised only one species of ophryoscolecid ciliate, Diploplastron affine, which is characterized by having two retractable ciliary zones in the anterior end of the body, two slender and juxtaposed skeletal plates on the right side, a rod shape macronucleus, and two contractile vacuoles. During study on the characterization of rumen ciliate community composition in Brazilian domestic sheep, we observed ciliates with atypical morphology but with diagnostic features of genus Diploplastron. This study describes Diploplastron dehorityi, a new species of ophryoscolecid ciliate, that differs from D. affine, primarily, in the morphology of skeletal plates, morphology of nuclear apparatus and body shape. In addition to the similarities between the new species and congener species, D. dehorityi has some morphological similarities to species of genus Eremoplastron.