Revisiting the taxonomy of Rhabdias fuelleborni Travassos, 1928 (Nematoda, Rhabdiasidae) with approaches to delimitation of species and notes on molecular phylogeny.

Rhabdias fuelleborni was described by Travassos (1926), who identified it in the lungs of Rhinella gr. marina (=Bufo marinus) from São Paulo. Later, Kloss (1971) added new information on the morphology and diagnosis of the species and new data on host and localities. However, its taxonomic status remains uncertain, due in particular to the morphological uniformity of Rhabdias spp., while the original description lacks important morphological data. In the present study, we found and reevaluated the type series of R. fuelleborni, as well as examining fresh material obtained from the type host and type locality, using light and scanning electron microscopy and molecular tools. The type series was deposited by Travassos (1926), and according to museum records the type locality is Paraty, Rio de Janeiro, Brazil. Our morphological and molecular analyses reinforce the taxonomic validity of this species, while molecular phylogeny identified a close relationship between R. fuelleborni and Rhabdias sp.4 and Rhabdias sp. from other studies, and from bufonids. Also R. fuelleborni are sister taxa with R. cf. stenocephala from Leptodactylidae from the northeast of Brazil and R. vencesi from Boophis madagascarariensis, Madagascar, according to mitochondrial COI gene.

Phylogenetic relationships of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) with descriptions of two new species.

Ochoterenatrema is a small genus of lecithodendriid digeneans that includes six species parasitic in Neotropical bats in North and South America. Members of this genus are characterized by the presence of a pseudogonotyl formed by thickened tegument on the left side of the ventral sucker. We examined morphology of specimens belonging to five species of Ochoterenatrema from Brazil, Ecuador and the USA and describe two new Ochoterenatrema spp. (Ochoterenatrema sphaerula n. sp. and Ochoterenatrema gracilis n. sp.). These new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles and the body shape. We have provided the first DNA sequences for Ochoterenatrema. Newly generated partial nuclear 28S rDNA and mitochondrial cox1 gene sequences were used to differentiate among species and study the phylogenetic affinities of Ochoterenatrema spp., including one of the new species, O. gracilis n. sp., as well as Ochoterenatrema diminutum, Ochoterenatrema fraternum and Ochoterenatrema cf. labda. The phylogeny of the Microphalloidea based on 28S had well-supported topology, particularly at the family level. The Ochoterenatrema clade was strongly supported; however, the internal topology of the clade was weakly supported. Comparison of sequences revealed 0.4-1.3% interspecific divergence in 28S and 9.1-19.7% interspecific divergence in cox1 among Ochoterenatrema spp. We hypothesize that extremely diverse fauna of bats in South and Central America harbors multiple undescribed species of Ochoterenatrema. Several lecithodendriid genera from bats and other hosts are yet to be included in future molecular phylogenetic analyses to test the monophyly of this extremely diverse digenean family and analyze evolutionary affinities of its constituent taxa.

A NEW SPECIES OF ALLINTOSHIUS (NEMATODA: HELIGMOSOMOIDEA) FROM TWO SPECIES OF BATS IN BRAZIL.

Allintoshius Chitwood, 1937 is the only genus of the family Ornithostrongylidae (Travassos, 1937) Durette-Desset and Chabaud, 1981 that parasitizes bats. Currently, there are 10 valid species in the genus, of which 3 were described from Brazil. This study describes a new species of Allintoshius and records the first occurrence of a nematode of this genus parasitizing Artibeus lituratus (Olfers). Allintoshius gomesae n. sp. is characterized by having anterior region coiled, cephalic vesicle with cuticular dilation striated transversely, and claviform esophagus. Synlophe in females consists of 16 cuticular ridges at the mid-body. Males have large caudal bursa, and conic and small spicules, and the gubernaculum is absent. Females have uterus didelphic, amphidelphic, tail tip tapered, and ovijector divided into 2 divergent branches, subequal in length. The new species differs from its congeners especially by the shape of the tail tip, vulvar opening, and size of spicules. Allintoshius gomesae is the fourth species of Allintoshius from Brazil and the first report in Ar. lituratus, increasing the number of species recognized of the genus.

HELMINTH FAUNA OF THECADACTYLUS RAPICAUDA (HOUTTUYN, 1782) (SQUAMATA, PHYLLODACTYLIDAE) IN THE CAXIUANÃ NATIONAL FOREST, FROM THE BRAZILIAN EASTERN AMAZON.

The Amazon region is an important region for biodiversity, and many new species are described for this region each year. However, the diversity of parasites infecting Amazonian reptiles is still poorly known, and there are few studies about how parasite communities of these hosts are structured. Our study was designed to elucidate the parasite community structure of Thecadactylus rapicauda, one of the largest species of gekkonid lizards in the world, and to discover the relative utility of size, sex, and body mass of Thecadactylus rapicauda as predictive factors of the abundance of helminth parasites. We analyzed 40 individuals of T. rapicauda, of which 34 had parasites (85% of prevalence), with 269 specimens allocated to 10 species. Body size of the host showed a positive influence on the abundance of helminths. Host sex is another important factor related to the abundance of nematodes: females were usually heavily infected. We conclude that T. rapicauda has a high richness of the helminth species and that most of the species found infecting this host were nematodes with a direct life cycle. Additionally, we found new helminth species, which should be described formally in the future.

Interrelationships of Anenterotrema (Digenea: Dicrocoeliidae) from Neotropical bats (Mammalia: Chiroptera) with description of a new species from Molossus molossus in Brazil.

Anenterotrema is a small genus of dicrocoeliids (Digenea: Dicrocoeliidae) containing 6 species found in Neotropical bats. Members of this genus are characterized by the lack of digestive system organs and, unlike the majority of dicrocoeliids, parasitize the intestine of their definitive hosts. In this study, we have morphologically examined newly collected specimens belonging to four species of Anenterotrema from Brazil, Ecuador, and Panama. Based on the data in original descriptions and our analysis of quality new specimens, we resurrected Anenterotrema freitasi, previously synonymized with A. eduardocaballeroi. We also described a new species of Anenterotrema from Molossus molossus in the Brazilian Amazon. The new species differs from congeners in several morphological features, most prominently in the size and structure of its highly muscular, protuberant ventral sucker. It is also characterized by the lack of the semi-circular thickening of the tegument around the oral sucker typical for some members of the genus. We used partial DNA sequences of the large ribosomal subunit gene (28S) and mitochondrial cytochrome c oxidase subunit 1 gene (cox1) to test the monophyly of Anenterotrema, and study the interrelationships and determine the inter- and intraspecific variation of three Anenterotrema spp. collected from different bat species in Brazil, Ecuador and Panama.

A New Species of Cosmocercoides (Ascaridida: Cosmocercidae) Parasitic in Tree Frogs from Southern Amazonia.

Cosmocercoides meridionalis n. sp. is described from the large and small intestines of 5 species of tree frogs belonging to the families Hylidae and Phyllomedusidae collected from 2 localities in Southern Amazonia. The new species is allocated to the genus Cosmocercoides Wilkie, 1930, due to the presence of papillae in rosettes, which are complex caudal papillae surrounded by punctuations. Cosmocercoides meridionalis n. sp. differs from its congeners by a combination of the following characters: the absence of the gubernaculum, the total length of the female (twice the size of the male), and the presence of 26 rosette papillae with a unique arrangement and distribution pattern: 11 pre-cloacal pairs, 1 ad-cloacal pair, and 1 post-cloacal pair. This is the first species of the genus described from the Amazonia region.

Phylogeny and systematics of the Proterodiplostomidae Dubois, 1936 (Digenea: Diplostomoidea) reflect the complex evolutionary history of the ancient digenean group.

The Proterodiplostomidae Dubois, 1936 is a relatively small family of diplostomoidean digeneans parasitising the intestines of reptilian hosts associated with freshwater environments in tropical and subtropical regions. The greatest diversity of proterodiplostomids is found in crocodilians, although some parasitise snakes and turtles. According to the most recent revision, the Proterodiplostomidae included 17 genera within 5 subfamilies. Despite the complex taxonomic structure of the family, availability of testable morphology-based phylogenetic hypotheses and ancient hosts, molecular phylogenetic analyses of the group were practically lacking. Herein, we use novel DNA sequence data of the nuclear lsrRNA gene and mitochondrial cox1 gene from a broad range of proterodiplostomid taxa obtained from crocodilian, fish, and snake hosts on four continents to test the monophyly of the family and evaluate the present morphology-based classification system of the Proterodiplostomidae in comparison with the molecular phylogeny. This first detailed phylogeny for the Proterodiplostomidae challenges the current systematic framework. Combination of molecular phylogenetic data with examination of freshly collected quality specimens and re-evaluation of morphological criteria resulted in a number of systematic and nomenclatural changes along with a new phylogeny-based classification of the Proterodiplostomidae. As the result of our molecular and morphological analyses: (i) the current subfamily structure of the Proterodiplostomidae is abolished; (ii) three new genera, Paraproterodiplostomum n. g., Neocrocodilicola n. g. and Proteroduboisia n. g., are described and Pseudoneodiplostomoides Yamaguti, 1954 is restored and elevated from subgenus to genus level; (iii) two new species, Paraproterodiplostomum currani n. g., n. sp. and Archaeodiplostomum overstreeti n. sp., are described from the American alligator in Mississippi, USA. Comparison of the structure of terminal ducts of the reproductive system in all proterodiplostomid genera did not support the use of these structures for differentiation among subfamilies (or major clades) within the family, although they proved to be useful for distinguishing among genera and species. Our study includes the first report of proterodiplostomids from Australia and the first evidence of a snake acting as a paratenic host for a proterodiplostomid. A key to proterodiplostomid genera is provided. Questions of proterodiplostomid-host associations parasitic in crocodilians are discussed in connection with their historical biogeography. Our molecular phylogeny of the Proterodiplostomidae closely matches the current molecular phylogeny of crocodilians. Directions for future studies of the Proterodiplostomidae are outlined.

A new species of Metadelphis Travassos, 1944 (Digenea: Dicrocoeliidae) from the gall-bladder of Molossus molossus (Pallas) (Chiroptera: Molossidae) in the Brazilian eastern Amazon.

Metadelphis tkachi n. sp. is described based on material from the gall-bladder of Molossus molossus (Pallas) (Chiroptera: Molossidae) in the Brazilian eastern Amazon. The main differentiating characters of the new species include lobed, tandem testes and elongated clusters of vitelline follicles commencing at the level of the seminal receptacle. The new species can be distinguished from Metadelphis lenti (Santos & Gibson, 1998) and Metadelphis apharyngotrema (Marshall & Miller, 1979) by the shape and position of the testes; from Metadelphis evandroi Travassos, 1944 by having tandem, lobed testes and shorter clusters of vitelline follicles; from Metadelphis compactus Travassos, 1955 by having tandem testes and more elongated clusters of vitelline follicles, and from Metadelphis alverangai Travassos, 1955 by having smaller testes and body, and vitellarium with large follicles. New morphological data are reported for M. lenti based on light and scanning electron microscopy; intraspecific variation was observed.

Oswaldocruzia lanfrediae n. sp. (Strongylida: Molineidae), a parasite of Leptodactylus paraensis Heyer (Anura: Leptodactylidae) in Brazil.

Leptodactylus paraensis Heyer, is a Neotropical anuran species that inhabits Rainforest habitats in the eastern Amazon, but because it has only been recently separated from the Leptodactylus pentadactylus (Laurenti) species group, little is known about its helminth fauna. This study describes a new species of Oswaldocruzia Travassos, 1917 and records the first occurrence of this genus parasitising L. paraensis and the second species for the Caxiuanã National Forest in the eastern Amazon, Brazil. Oswaldocruzia lanfrediae n. sp. is characterised by having an anterior extremity with a smooth cephalic vesicle divided into two portions, a claviform oesophagus, well-developed cuticular longitudinal ridges and lateral alae. Females have a well-developed ovojector, with didelphic and amphidelphic uteri. Males show complex robust spicules divided into a slightly curved shoe, a bifurcated fork and a blade terminating in 2-3 processes. The new species differs from its congeners especially regarding the lateral alae and the morphology of the spicules, in addition to morphometric characters such as body size, oesophagus length, deirid position, nerve-ring position and relative position of the vulva in females.

Description, biology and molecular characterisation of Serpentirhabdias moi n. sp. (Nematoda: Rhabdiasidae) from Chironius exoletus (Serpentes: Colubridae) in Brazil.

A new species of the genus Serpentirhabdias Tkach, Kuzmin et Snyder, 2014, S. moi n. sp., is described from a colubroid snake Chironius exoletus from Caxiuanã National Forest, State of Pará, Brazil. The species is characterised by having a triangular oral opening, absence of the buccal capsule, presence of six minute onchia in the oesophastome, and excretory glands of approximately the same length as the oesophagus. These qualitative morphological characters, as well as some measurements, differentiate the new species from other Neotropical and Nearctic Serpentirhabdias spp. The morphological description of parasitic adults of S. moi n. sp. is complemented by the description of free-living stages including males, females, and infective larvae. Comparative analysis of partial sequences of cox1 and 12S mitochondrial genes strongly supported the status of S. moi n. sp. as a new species. Molecular phylogeny based on sequences of the nuclear DNA region spanning the 3' end of the 18S nuclear rRNA gene, ITS region (ITS1 + 5.8S + ITS2) and 5' end of the 28S gene supported monophyly of all rhabdiasid genera included in the analysis and placed the new species into the Serpentirhabdias clade as sister taxon to S. fuscovenosa.

Neocosmocercella fisherae n. sp. (Nematoda: Cosmocercidae), a parasite of the large intestine of Phyllomedusa bicolor (Boddaert) (Anura: Phyllomedusidae) from the Brazilian Amazon.

Neocosmocercella fisherae n. sp. is the first nematode species found parasitising Phyllomedusa bicolor from the Brazilian Amazon Region. The new species has a triangular oral opening, with bi-lobed lips, and is distinguished from N. bakeri (triangular oral opening with simple lips), and from N. paraguayensis (hexagonal oral opening with bi-lobed lips). Additionally, the new species has ciliated cephalic papillae, which are absent in the other species of the genus. The reduced uterine sac and the presence of a single egg in the uterus in females are the main morphological characters that differentiate the new species from its congeners N. bakeri (8-10 eggs) and N. paraguayensis (10 eggs, based on the allotype). Additionally, the new species differs from the other two species of the genus by morphometric characters such as the size of spicules and gubernaculum in males and the vagina in females. Until now, phyllomedusid anurans are the only known hosts for the nematodes of this genus. The present work describes the third species of the genus and the first species of nematode parasitising P. bicolor.

Gongylonema Parasites of Rodents: A Key to Species and New Data on Gongylonema neoplasticum.

Specimens of the genus Gongylonema were collected from the gastric mucosa of rodents of Rattus rattus Linnaeus, 1758, and Rattus norvegicus Berkenhout, 1769, collected in urban areas in Belém, Pará, in the eastern Brazilian Amazon. The helminths were processed for analysis using light microscopy and scanning electron microscopy (SEM) techniques and presented taxonomic characteristics of the species Gongylonema neoplasticum. The SEM analyses revealed the presence of 2 developed buccal plates (1 dorsal, 1 ventral), also called interlabia, with a prominent and bifurcated ventral plaque. The occurrence of the bifurcated ventral interlabium had not yet been identified by any other author from G. neoplasticum. As a result of our extensive research on published data on Gongylonema spp., we propose a taxonomic key for species of this genus that parasitize rodents. This is the first record of G. neoplasticum in urban areas of the Brazilian Amazon.

Redescription of Oswaldocruzia chambrieri (Strongylida: Molineidae) from Rhinella margaritifera (Anura: Bufonidae) in Caxiuanã National Forest, Brazil.

Oswaldocruzia chambrieri Ben Slimane et Durette-Desset, 1993 is redescribed from specimens collected from the small intestine of the South American common toad, Rhinella margaritifera, from Caxiuanã National Forest in Pará, Brazil, using light and scanning microscopy and molecular analysis of Cytochrome Oxidase I (COI) - coding regions of DNA. The discovered nematodes are characterized by a type III caudal bursa with two papillae, rays 4 with a median groove, and spicules divided into a blade, a shoe and a fork. Cervical alae are absent, the cephalic vesicle is divided into two portions, and the synlophe has low ridges without chitinous supports. The present study establishes the Caxiuanã National Forest as a new location for O. chambrieri, which had previously been reported as a parasite of R. margaritifera in Ecuador, uses light microscopy and scanning electron microscopy (SEM) to identify new morphological characters of the species and represents the second molecular sequence deposited for the Oswaldocruzia genus.

Rhinella marina (Amphibia: Bufonidae) Versus Rhabdias paraensis (Nematoda: Rhabdiasidae): Expanding the View on a Natural Infection.

Amphibian and reptile lungs are frequently infected with Rhabdias parasites, and this condition ultimately leads to reduced survival, performance, and growth because of granulomatous inflammation, nodule formation, and nematodal pneumonia onset. Here we investigate the histopathological features of naturally infected Rhinella marina by the lung nematode Rhabdias paraensis. A total of 10 host animals were captured in peridomiciliar areas in the eastern Brazilian Amazon, and anatomic-histological analyses were performed on both the infected and non-infected lungs of these amphibians. Helminths were usually found within the secondary and primary septa of infected lungs whereas parasites were not detected within vessels or adhering to tissues. In addition, we observed discrete erythrocytes, diapedesis foci, few granulocytes and erythrocytes in the interseptal spaces, discrete cell infiltration, and a small number of melanomacrophages, and no granulomas or cysts were observed. New aspects related to changes in tissue and helminth-host interactions are discussed for the relationship of R. paraensis × Rhi. marina from the Amazon region.

Strongyluris amazonicus n. sp. (Nematoda: Heterakidae): a parasite of Tropidurus oreadicus from the Brazilian Amazon.

Tropidurus oreadicus Rodrigues 1987 is a recently described species, thus there are no reports of helminthes parasites for this species. In this study, a morphological characterisation was performed of a nematode species parasite of the large intestine of T. oreadicus captured in an urban area. This urban area is similar to the estuarine dale from Guamá river and the "igarapés" of Belém, Pará State, Brazil. Morphological analysis suggested that the parasite is a new species of nematode, based mainly on the number and distribution pattern of the caudal papillae in males, which is unique for this species. In the present work, we describe for the first time the structure, number, and disposition of the cervical papillae. The morphological data were supported by scanning electron microscopy, which served as an important tool for distinguishing these nematodes from other species of the genus.