Morphological and molecular characterization of Cosmocercoides amapari n. sp. (Nematoda: Cosmocercidae), parasitic in hylid frogs from the Brazilian Amazon.

Cosmocercoides Wilkie, 1930 are gastrointestinal parasites commonly found in amphibians and reptiles, with 4 species reported from the Neotropical region. In the present study, a new species of Cosmocercoides, namely Cosmocercoides amapari n. sp. is described using integrated approaches such as light and scanning microscopy, and DNA sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene. The specimens were collected from the large intestine of 3 species of hylid frogs in Amapá, Brazil. The new species can be distinguished from its congeners by morphological traits, including the pattern of caudal papillae, absence of the gubernaculum, 2 poorly sclerotized spicules, presence of lateral alae and somatic papillae along the body. In addition, our molecular analyses and phylogenetic reconstructions strongly support the status of the new taxon, which clustered poorly with a large clade of Cosmocerca spp. Cosmocercoides amapari n. sp. is the 29th species of the genus, the 5th from the Neotropical region, the third reported in Brazil, the second described from the Amazon region and the first belonging to the Neotropical region with molecular data.

Vogtnema asymmetrica n. gen., n. sp. (Nematoda: Cosmocercoidea: Atractidae) parasite of Rhinoclemmys punctularia (Daudin) (Testudines: Geoemydidae) from eastern Amazon, Brazil.

Nematodes collected from the stomach and large intestine of Rhinoclemmys punctularia (Daudin) from the eastern Amazon, Brazil, are assigned to a new genus, Vogtnema n. gen., allocated in the family Atractidae (Cosmocerdoidea). Vogtnema n. gen. differs from all other genera of Atractidae based on the combination of the following morphological characters: four lips V-shaped (two subdorsal and two subventral) without sclerotized structures associated, absence of sclerotized formations and/or pieces surrounding the oral opening, oral aperture surrounded by flange internal membranous structures; in males the larger spicule is alate. The type species Vogtnema asymmetrica n. gen., n. sp. has the five precloacal pairs of papillae situated on the right side of the body that are more prominent than those on the left side. We also provide an identification key to the genera of the family Atractidae.

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 Rhabdias (Nematoda: Rhabdiasidae), a lung parasite of Pristimantis chiastonotus (Anura: Strabomantidae) from the Brazilian Amazon: description and phylogenetic analyses.

Rhabdias Stiles and Hassal, 1905 comprises approximately 90 species of parasitic lung nematodes of amphibians and reptiles that have a wide distribution, with 21 species occurring in the Neotropics. In the present study, we describe Rhabdias waiapi n. sp. found parasitizing the lungs of the anuran species Pristimantis chiastonotus from the Amazon Biome in the Amapá State, Northern Brazil. The new species is characterized by having an elongated body, expansions of the cuticular inflation in the anterior end that become more discrete along the body, an anterior end with a slight constriction at the level of the esophageal apex with four rounded subapical elevations of the body wall, six lips, four near the edge of the oral opening and two more distant lateral ones, and a gradually tapering elongated tail. In addition, molecular analyses and phylogenetic reconstructions were made, with sequences from the coding region of the mitochondrial cytochrome c oxidase subunit I gene. Those results strongly support the status of the new taxon, which formed a poorly supported clade with Rhabdias sp. 5 from Anolis brasiliensis from Northeast Brazil. Rhabdias waiapi n. sp. is the 19th species of the genus described in the Neotropics for amphibians, the 10th in Brazil, the second described from hosts of the family Strabomantidae from the Neotropical region, and the first amphibian nematode species described in the Amapá State.

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.

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.

First report of filarial nematodes in free-living pitheciid primates.

Although little studied, infections with nematodes of the Onchocercidae Leiper, 1911, predominated by the genera Dipetalonema Diesing, 1861 and Mansonella Faust, 1929, are frequent in wild primates and human populations in the Neotropical forest areas. This study reports natural infections with Dipetalonema freitasi Bain, Diagne & Muller, 1987 and D. gracile (Rudolphi, 1809) in two free-living species of pitheciid primates, extending the known geographical distribution of these species to the forest of the Peruvian Amazon. Adult worms were recovered from the thoracic and abdominal cavities of two species of monkeys, Pithecia monachus monachus (É. Geoffroy Saint-Hilaire) and Cacajao calvus ucayalii (Thomas) (Primates: Pitheciidae), collected along the Yavari-Mirin River basin and analysed via light and scanning electron microscopy. Both host species represent new host records for D. freitasi and D. gracile. Morphometric data are also presented for the sampled filarial worms in addition to morphological details obtained through light and electron microscopy examination of D. freitasi specimens.

A new species of Procamallanus Baylis, 1923 (Nematoda, Camallanidae) from Astronotusocellatus (Agassiz, 1831) (Perciformes, Cichlidae) in Brazil.

A new species of Procamallanus Baylis, 1923 was found as a parasite of the fish Astronotusocellatus (Agassiz, 1831) from a lake in the Jardim Botânico Bosque Rodrigues Alves, Belém, Brazil. Procamallanusspiculastriatus sp. n. has a smooth buccal capsule and a well-developed basal ring that is armed with four sclerotized tooth-like structures. The male of the new species is similar to the two species that are known from Brazilian fish, P.peraccuratus Pinto, Fábio, Noronha & Rolas, 1976, and P.annipetterae Kohn & Fernandes, 1988, by the absence of the gubernaculum. It differs from these two by the morphology of the buccal capsule, the number are arrangement of the caudal papillae in males, the size and morphology of the spicules and the shape of the tail of both sexes. Procamallanusspiculastriatus sp. n. is the third species discovered in fish from Brazil. This finding extends the geographical distribution of the genus into the Brazilian Amazon.

Canine filariasis in the Amazon: Species diversity and epidemiology of these emergent and neglected zoonoses.

Dirofilaria immitis and Acanthocheilonema reconditum are common parasites in dogs but have also been reported parasitizing humans. The differential diagnosis and epidemiological evaluation of these zoonoses are important to the development of efficient public health policies and control strategies. The purpose of this study was to carry out an epidemiological survey by using molecular methods for the specific identification of filarid parasites of domestic dogs in the Marajó mesoregion, State of Pará (PA), Brazil. A total of 418 canine blood samples from Marajó mesoregion (Northern Brazil) were collected, submitted to DNA extraction, polymerase chain reaction (PCR) with "pan filarial" primer, subsequent sequencing and sequence analysis using BLASTn software comparison with previously deposited sequences in GenBank. After that, a phylogenetic analysis by Maximum Parsimony was performed to aid the specific diagnosis. The obtained sequences showed the occurrence of 9 (2.15%) dogs infected with D. immitis and 30 (7.18%) by A. reconditum, with a confidence interval of 95%, there were no cases of co-infection. We observed that male dogs were more likely to D. immits and A. reconditum infection. However, age was not significant to both infections. This study reports for the first time the occurrence of A. reconditum in the northern region of Brazil and confirmed the presence of D. immitis in the Marajó mesoregion.

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.

A new species of Neocosmocercella Baker & Vaucher, 1983 (Nematoda: Cosmocercidae), a parasite of Phyllomedusa vaillantii Boulenger (Anura: Phyllomedusidae) in the Caxiuanã National Forest, eastern Amazon, Brazil.

Neocosmocercella bakeri n. sp. is described from the large intestine of Phyllomedusa vaillantii Boulenger collected in the Caxiuanã National Forest in the Eastern Brazilian Amazon. The new species is easily distinguished from the type-species of the genus, Neocosmocercella paraguayensis Baker & Vaucher, 1983 in possessing a triangular mouth opening with three simple lips (vs three bi-lobed lips and hexagonal mouth opening) and somatic papillae, which are absent in N. paraguayensis. The males of the new species are distinguished by the distribution of the sessile cloacal papillae and the dimensions of the gubernaculum, whereas the females are distinguished by their smaller size and opisthodelphic uterus. This study expands the diagnostic characters of Neocosmocercella Baker & Vaucher, 1983, reports the first species parasitising anurans of the Brazilian Amazon, a new host record for the genus, and the description of the second species of the genus.

Gene silencing of Dim-1, a member of the disorganized muscle family, in Haemonchus contortus.

RNA interference has been widely used in parasites. In Haemonchus contortus, reproducible silencing has been reported; however, in this species an altered phenotype has been observed for only a few genes silenced by RNA interference. The aim of this work was to evaluate the effect of silencing Dim-1, a member of the disorganized muscle family on third stage larvae (L3) of H. contortus. Effective silencing of Dim-1 in L3 led to reduced L3 migration and slowed larval development from L3 to early L4. To our knowledge, this is the first report to demonstrate Dim-1 silencing affecting phenotypic characteristics of this parasite.

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.

Acanthocephala Larvae parasitizing Ameiva ameiva ameiva (Linnaeus, 1758) (Squamata: Teiidae).

Knowledge concerning the taxonomy and biology of species of Acanthocephala, helminth parasites of the helminth species of the phylum Acanthocephala, parasites of lizards in Brazilian Amazonia, is still insufficient, but reports of Acanthocephala in reptiles are becoming increasingly common in the literature. Cystacanth-stage Acanthocephalan larvae have been found in the visceral peritoneum during necropsy of Ameiva ameiva ameivalizards from the "Osvaldo Rodrigues da Cunha" Herpetology Collection of the Emílio Goeldi Museum, Belém, Pará, Brazil. The aim of this study was to present the morphological study of the Acanthocephala larvae found in A. ameiva ameiva lizard.

The morphology of free-living stages and immature parasites of Rhabdias paraensis (Nematoda: Rhabdiasidae), a parasite of Rhinella marina (Anura: Bufonidae) in Brazil.

Rhabdias paraensis Santos, Melo, Nascimento, Nascimento, Giese et Furtado, 2011 was described based on fully gravid worms. Further investigations on the free-living stages, immature worms and young individuals were facilitated by cultivation in the laboratory, which allowed us to add new information about the morphology and development of the species. Observations on the free-living development of R. paraensis showed that the life cycle is typical of Rhabdias, with alternation of gonochoristic and hermaphroditic generations and without homogony. Males of the free-living generation were different from those in several species of the genus studied previously. In the original description, the excretory glands and duct were absent in gravid specimens of R. paraensis, while in this study, distinct excretory glands and a duct were observed in immature and young individuals. Additionally, we recognised the separation of the buccal capsule walls into anterior and posterior portions and described the specific shapes of these portions in lateral and apical view. Studies on the morphology and development of free-living stages of Rhabdias spp. from Neotropical regions may provide additional information for species determination.

Description of Serpentirhabdias atroxi n. sp. (Nematoda: Rhabdiasidae), a parasite of Bothrops atrox (Linnaeus) (Reptilia: Serpentes: Viperidae) in Brazilian Amazonia.

A new lung-dwelling nematode species is described from the common lancehead Bothrops atrox (Linnaeus) in the Brazilian Amazon Region. The species is assigned to the genus Serpentirhabdias Tkach, Kuzmin & Snyder, 2014 based on the presence of six lips arranged in two lateral groups, the absence of prominent cuticular inflations, and lung parasitism in snakes. Serpentirhabdias atroxi n. sp. differs from other species of the genus mainly by details of the morphology of the anterior end: cuticularised ring surrounding the anterior part of the buccal cavity and six minute onchia present in the oesophastome. Serpentirhabdias atroxi n. sp. is the seventh species of the genus known from the Neotropical Realm and the second species described from viperid snakes.