Morphology and ASAP analysis of the important zoonotic nematode parasite Baylisascaris procyonis (Stefahski and Zarnowski, 1951), with molecular phylogenetic relationships of Baylisascaris species (Nematoda: Ascaridida).

Species of Baylisascaris (Nematoda: Ascarididae) are of great veterinary and zoonotic significance, owing to cause Baylisascariosis or Baylisascariasis in wildlife, captive animals and humans. However, the phylogenetic relationships of the current 10 Baylisascaris species remain unclear. Moreover, our current knowledge of the detailed morphology and morphometrics of the important zoonotic species B. procyonis is still insufficient. The taxonomical status of B. procyonis and B. columnaris remains under debate. In the present study, the detailed morphology of B. procyonis was studied using light and scanning electron microscopy based on newly collected specimens from the raccoon Procyon lotor (Linnaeus) in China. The results of the ASAP analysis and Bayesian inference (BI) using the 28S, ITS, cox1 and cox2 genetic markers did not support that B. procyonis and B. columnaris represent two distinct species. Integrative morphological and molecular assessment challenged the validity of B. procyonis, and suggested that B. procyonis seems to represent a synonym of B. columnaris. Molecular phylogenetic results indicated that the species of Baylisascaris were grouped into 4 clades according to their host specificity. The present study provided new insights into the taxonomic status of B. procyonis and preliminarily clarified the phylogenetic relationships of Baylisascaris species.

Oxysomatium brevispiculum n. sp. (Nematoda: Cosmocercidae) from Amphisbaena alba Linnaeus (Squamata, Amphisbaenidae) in the Cerrado Biome from Brazil.

In the present study, we describe Oxysomatium brevispiculum n. sp. (Ascaridida: Cosmocercidae) parasitizing Amphisbaena alba Linnaeus (Squamata: Amphisbaenidae) in the municipality of Uberlândia, Cerrado Biome, Minas Gerais, Brazil. Oxysomatium brevispiculum n. sp. differs from its congeners by having shorter spicules and by the number and arrangement of caudal papillae. The males of the new species have a precloacal unpaired papilla and can be easily distinguished from O. caucasicum in which this morphological trait is absent. Oxysomatium brevispiculum n. sp. differs from the other three species of the genus by the number and arrangement of caudal papillae, with 13 pairs + 1 unpaired precloacal papilla, arrangement 8+1:2:3, while O. brevicaudatum has 14-16 pairs +1 unpaired papilla, and 7-9+1:1:6 arrangement, O. petrolinensis has 16 pairs +1 unpaired papilla, and 8+1:1:7 arrangement, and O. dollfusi with 15-16 pairs +1 unpaired papilla, and 8-9+1:1:6 arrangement. Oxysomatium brevispiculum n. sp. is the fifth species of this genus, the second species in a Neotropical host, and the first species of this genus described in amphisbaenid hosts. In addition, the present study provides an identification key for the species of this genus.

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.

Austraplectana extranes sp. n. (Nematoda: Cosmocercoidea) from Australian amphibians Mixophyes carbinensis Mahony, Donnellan, Richards & McDonald (Anura: Myobatrachidae) and Rhinella marina (Linnaeus) (Anura: Bufonidae).

The genus Austraplectana Baker, 1981 is a poorly known group of cosmocercoid nematodes. In the present study, a new species of Austraplectana, A. extranes sp. n., was described using both light and scanning electron microscopy, based on specimens collected from the carbine barred frog Mixophyes carbinensis Mahony, Donnellan, Richards & McDonald (Anura: Myobatrachidae) and the cane toad Rhinella marina (Linnaeus) (Anura: Bufonidae) in Australia. Austraplectana extranes sp. n. can be easily distinguished from its congener by the much longer and different morphology of tails in both sexes, the different number and arrangement of caudal papillae, the presence of single precloacal median heart-shaped papilla and large posterior protuberance with cuticular comb-like fringe in male.

Cosmocerca goroensis n. sp. (Nematoda: Cosmocercidae) from South Africa and its phylogenetic relationships with other cosmocercids based on partial 28S sequences.

Nematodes of the genus Cosmocerca are commonly found in various amphibians in South Africa and in most cases are identified as C. ornata. However, after detailed morphological studies and molecular approaches, three new species of the genus were recently described from three different frogs in South Africa. In present study, we describe another new species - Cosmocerca goroensis parasitising the Northern Pygmy Toad Poyntonophrynus fenoulheti in Soutpansberg mountains, Limpopo province, South Africa. The new species is characterised by prominent sex dimorphism, wide lateral alae, numerous somatic papillae in both sexes, and wide triangularly shaped gubernaculum and simple prominent spicules in males. Cosmocerca goroensis n. sp. distinguished from congeners, previously reported in Southern Africa by the shape of the gubernaculum and arrangement of somatic papillae in males. Morphological differences were confirmed by molecular analysis based on fragments of the 28S gene. Phylogenetic analysis based on the 28S gene fragments, including C. goroensis n. sp. and newly obtained sequence of C. ornata from Pelophylax lessonae from Ukraine, supported previously known data of closer relationships between species of Cosmocerca and Aplectana and more distant with Cosmocercoides spp.

Orientatractis bowseri n. sp. (Nematoda: Cosmocercoidea: Atractidae) parasite of Peltocephalus dumerilianus (Schweigger) (Testudines: Podocnemididae) from the Brazilian Amazon.

Peltocephalus dumerilianus (Schweigger) is a diurnal freshwater turtle widely distributed in the Orinoco and Amazon River basins in Brazil, Colombia, Peru, Ecuador, Venezuela, and French Guiana. During a helminthological survey of freshwater turtles from the Brazilian Amazon, numerous nematodes with characteristics of the genus Orientatractis Petter, 1966 were recovered from the stomach of Peltocephalus dumerilianus. These nematodes were different from all congeneric species, and we describe herein. Orientatractis bowseri n. sp. differs from all known species of the genus by having dorsal cuticular projections on the posterior region of the females, distribution of caudal papillae and by having papilla-like ornamentations on the cuticle distributed from the mid-esophagus to the mid portion of the body. This study represents the fourth specie of the genus Orientatractis reported in Brazil, the seventh species of Orientatractis in the Neotropical region, and the third found infecting freshwater turtles in Brazil.

Further studies on the morphology of Baylisascaris transfuga (Rudolphi, 1819) (Nematoda: Ascaridomorpha: Ascarididae) from the polar bear Ursus maritimus Phipps (Carnivora: Ursidae).

Baylisascaris transfuga (Rudolphi, 1819) is a common parasitic nematode in the digestive tract of various species of bears worldwide, with great veterinary significance. However, our present knowledge on the morphology of B. transfuga remains insufficient. In the present study, the detailed morphology of B. transfuga was studied using light and scanning electron microscopy (SEM), based on specimens collected from the polar bear Ursus maritimus Phipps (Carnivora: Ursidae) in the Shijiazhuang Zoo, China. The results revealed some morphological and morphometric variation between the present specimens and some of those from previous studies, including oesophageal length of female, number and morphology of postcloacal papillae and morphology of tail of males. Present SEM observations clearly showed the detailed morphology of lips, cervical alae, cloacal ornamentation, precloacal medioventral papilla, phasmids and tail tip. These supplementary morphological and morphometric data enable us to identify this ascaridid nematode more accurately.

Two new species of Dichelyne Jägerskiöld, 1902 (Nematoda: Cucullanidae) parasitising marine fishes in Iraq.

Recent examinations of some marine fishes from off the southern coast of Iraq revealed the presence of two new species of Dichelyne (Dichelyne) Jägerskiöld, 1902 (Nematoda: Cucullanidae), both intestinal parasites, namely: D. (D.) lethrini n. sp. from the smalltooth emperor Lethrinus microdon Valenciennes (Lethrinidae, Acanthuriformes) and D. (D.) iraqiensis n. sp. from the humphead snapper Lutjanus sanguineus (Cuvier) (Lutjanidae, Acanthuriformes). Specimens are described and illustrated based on light and scanning electron microscopical examinations. Dichelyne (D.) lethrini n. sp. is mainly characterised by the length of spicules (522 µm), presence of a dorsal caecum, location of deirids at the level of the posterior end of oesophagus and the post-oesophageal position of the excretory pore, number (10 pairs) and distribution of caudal papillae in the male, and the presence of two minute terminal spines on the female tail tip. Dichelyne (D.) iraqiensis n. sp. differs from the species of the subgenus Dichelyne mainly based on the absence of caudal spines, the presence of a dorsal caecum, the length of spicules (789-884 µm), the location of deirids and the excretory pore approximately at the same level, and the size of eggs (72-78 × 42-51 µm). These new species infecting marine fishes in Iraq represent new geographic records of cucullanids of the subgenus Dichelyne from fishes in marine waters off Iraq.

Population genetic structure and morphological diversity of Cruzia tentaculata (Nematoda: Ascaridida), a parasite of marsupials (Didelphinae), along the Atlantic Forest on the eastern coast of South America.

Cruzia tentaculata is a helminth parasite of marsupials and has a wide geographic distribution from Mexico to Argentina. The aim of this study was to analyse the genetic population structure of this nematode along the Atlantic Forest biome. Cruzia tentaculata specimens were recovered from Didelphis aurita, Didelphis albiventris and Philander quica in 9 localities. Morphological and morphometric data were investigated for phenotypic diversity among localities and hosts using multivariate discriminant analysis of principal components. Phylogenetic relationships of C. tentaculata were determined using maximum likelihood and Bayesian inference. The population structure was analysed by fixation indices, molecular variance analysis, Tajima's D and Fu's Fs neutrality tests, Mantel tests and Bayesian clustering analysis. A higher significant morphometric difference for males was observed between localities. In the haplogroup networks, 2 groups were recovered, separating locations from the north and from the south/southeast. The morphometric variation in C. tentaculata between different localities was compatible with this north and southeast/south pattern, suggesting adaptation to different ecological conditions. Population genetic analyses suggested a pattern of evolutionary processes driven by Pleistocene glacial refugia in the northeast and southeast of the Atlantic Forest based on the distribution of genetic diversity.

Three new species of Cosmocerca Diesing, 1861 (Nematoda: Cosmocercidae) parasitising frogs Cacosternum boettgeri Boulenger, 1882, Kassina senegalensis Dumeril and Bibron, 1841 and Phrynomantis bifasciatus Smith, 1847 from South Africa.

Cosmocercid nematodes have been documented with much criticism due to the numerous inaccurate descriptions, redescriptions and synonymisation of found species. This is due to indistinguishable characters of females and the lack of male specimens found. Consequently, the species C. ornata is the most commonly found species worldwide and the only species of the genus reported in South Africa. In the present study, we found Cosmocerca in three different amphibian species, namely Cacosternum boettgeri, Kassina senegalensis and Phrynomantis bifasciatus. Based on differences in the shape of the gubernaculum and number of papillae, the found nematodes were assigned to three new species namely C. daly n. sp., C. monicae n. sp. and C. makhadoensis n. sp. Descriptions of species are followed by pairwise and phylogenetic analysis of partial ITS-28S sequences. All three species were found only in their host types from distant localities. Therefore, we hypothesise that host specificity of Cosmocerca from South African amphibians might be rather high and that the presence of C. ornata throughout South Africa is rather doubtful.

Subulura eliseae sp. n. (Ascaridida: Subuluroidea), a parasite of Marmosa spp. from Amazon rainforest, Brazil.

The parasite biodiversity of mouse opossums in Brazil remains incompletely explored. We describe a new species of Subulura (Ascaridida: Subuluroidea) from the large intestine of the white-bellied woolly mouse opossum, Marmosa constantiae, based on the results of light and scanning electron microscopy (SEM). We also partially sequenced the mitochondrial cytochrome c oxidase I (MT-CO1) gene of the new species, using molecular phylogenetic analyses to determine its relationships within the Subuluroidea superfamily. As molecular data on subuluroid species are extremely limited, few inferences could be drawn from our phylogenies. Our SEM observations showed the detailed morphology of the cephalic extremity, precloacal pseudo-sucker, caudal papillae, phasmids and vulva. Subulura eliseae sp. n. differs from the other four Subulura parasites species of marsupials by the number of caudal papillae and the structure dimensions, and size of the spicule. Moreover, S. eliseae sp. n. has ten pairs of caudal papillae, which is unique compared to other species. We present morphometric and molecular data on this new species, contributing to future studies on subuluroids.

First molecular assessment on Cosmocerca spp. from Brazilian anurans and description of a new species of Cosmocerca (Ascaridomorpha: Cosmocercoidea) from the white-spotted humming frog Chiasmocleis albopunctata (Boettger, 1885) (Anura: Microhylidae).

Cosmocerca spp. are common nematode parasites of amphibians. We provide herein molecular data for two species of Cosmocerca and describe a new species, Cosmocerca albopunctata n. sp., using light microscopy and molecular data (cytochrome c oxidase subunit 1 - COI mtDNA). Cosmocerca albopunctata n. sp. can be easily distinguished from other congeneric species by the combination of characteristics such as body size, length of spicules and gubernaculum, and the arrangements and number of caudal papillae (7 + 1:1 + 1:6). The phylogenetic results based on the partial COI mtDNA sequences clustered the new species in a monophyletic clade along with the other sequences of Cosmocerca spp. Therefore, our results contribute to the knowledge about the species diversity and genetic data for Cosmocerca spp. in the Neotropical region.

Endoparasite community structure of an anuran assemblage in the Caatinga, Northeastern Neotropical Region.

Amphibians are a widespread Chordata taxon and are important for maintaining the balance of both terrestrial and aquatic ecosystems. Brazil has a rich amphibian fauna; however, little is known about the role of their ecology and phylogenetic relationships during the assembly processes of associated endoparasite communities. Herein, we describe an endoparasite community in an anuran assemblage in the Caatinga, a unique biome of dry forests in north-eastern Brazil. We studied endoparasite diversity, as well as the effects of body length, body mass, body volume and sex on parasite abundance. We also investigated the influence of ecological and historical factors and anuran microhabitat use on endoparasite composition. We analysed individuals from 13 anuran species distributed across five families: Odontophrynidae (Proceratophrys cristiceps); Leptodactylidae (Leptodactylus fuscus, Leptodactylus vastus, Leptodactylus macrosternum, Leptodactylus troglodytes and Physalaemus cuvieri); Hylidae (Pithecopus gonzagai, Scinax x-signatus, Boana raniceps and Dendropsophus nanus); Bufonidae (Rhinella diptycha and Rhinella granulosa); and Microhylidae (Dermatonotus muelleri). We found nine species of endoparasites, including seven nematodes (Aplectana membranosa, Cosmocerca sp., Oswaldocruzia mazzai, Raillietnema spectans, Rhabdias fuelleborni, Schrankiana sp. and Physaloptera sp.), one species of Trematoda (Glypthelmins pseudium) and one non-identified cestode. There was no significant relationship between endoparasite abundance and host body length, body mass, body volume and sex. A phylogenetic principal component analysis showed that ecological factors had a greater influence on endoparasite assemblage than historical factors. Similarly, our results showed that ecological factors had a greater influence on anuran microhabitat use compared to historical factors, which contributed to the generalist characteristics presented by most of the sampled endoparasite species.

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.

A new species of nematode parasite, Cucullanus tunisiensis sp. nov. (Nematoda: Cucullanidae) from Epinephelus aeneus (Perciformes: Serranidae) off Tunisia.

Cucullanus tunisiensis sp. nov., (Nematoda: Cucullanidae), collected from the intestine of the white grouper Epinephelus aeneus from waters off the coast of Tunisia is described based on light and scanning electron microscopic observations. The new species is characterized by the presence of lateral alae, ventral sucker, long unequal spicules (left spicule 2474-2789 µm long, right spicule 2357-2518 µm long). This is the sixth nominal species of the genus Cucullanus Müller, 1777 and the first representative of this genus infecting fishes of Serranidae family reported from Tunisian waters.

Cucullanus bulbosus (Lane, 1916) Barreto, 1918 (Nematoda, Cucullanidae) from the common ponyfish Leiognathus equulus (Leiognathidae): Morphology and molecular study.

The common ponyfish Leiognathus equulus is a marine fish species with very high commercial value. Little information is available about its parasitic infections. Based on light and scanning electron microscopy, as well as sequencing and analysis of the partial regions of the ITS-1, 18S rRNA, COX1 genes, were employed for the systematic evaluation of a nematode parasite, which it first isolated from L. equulus in Jeddah Province, Red Sea, Saudi Arabia. Results revealed that this nematode parasite closely resembles the previously described Cucullanus bulbosus. Microscopic examination showed that it distinguished from congeners by the unique structure of hemispherical elevation at pseudobuccal capsule level, the ratio of esophagus/body length, spicules size, presence of pre-cloacal sucker, rod-shaped gubernaculum, and the arrangement of caudal papillae in males. Phylogenetic analyses based on ITS-1, 18S rRNA, and COX1 gene regions were constructed to investigate phylogenetic relationships between this parasite species and other related taxa. Results supported that Cucullanus bulbosus resembles a sister of Cucullanus genypteri, Cucullanus pulcherrimus, Cucullanus bourdini, Cucullanus extraneus, and Cucullanus hainanensis by using different genetic markers. This study provides more information about combining morphological and molecular data to identify Cucullanus species with the first natural occurrence in the common ponyfish inhabited in Saudi Arabia.

Integrated evidence reveals a new species of Cosmocerca (Ascaridomorpha: Cosmocercoidea) from the Asiatic toad Bufo gargarizans Cantor (Amphibia: Anura).

Species of Cosmocerca Diesing, 1861 (Ascaridomorpha: Cosmocercoidea), are common nematode parasites of amphibians. In the present study, a new species of Cosmocerca, namely C. simile n. sp., was described using light and scanning electron microscopy, and sequencing different nuclear and mitochondrial genetic markers (i.e. small ribosomal DNA (18S), large ribosomal DNA (28S), internal transcribed spacer (ITS) and cytochrome c oxidase subunit 1 (cox1)). Cosmocerca simile n. sp. differs from its congeners based on body size, morphology and number of plectanes, relative length of spicules and gubernaculum and spicules to total body length and morphology and length of tail. Molecular analysis showed no nucleotide polymorphisms among different individuals of the new species regarding nuclear DNA. Very low intraspecific nucleotide variation (0.52-0.78%) was detected in cox1 mtDNA. In contrast, the level of interspecific nucleotide variation between C. simile n. sp. and its congeners were distinctly higher (2.74-18.1% in the partial ITS region and 10.2-13.5% in the partial cox1 region, respectively) than that of intraspecific variation. Phylogenetic analyses using maximum likelihood (ML) inference based on the partial ITS and cox1 sequence data both supported the new species to be a member of the genus Cosmocerca, and formed a sister relationship to C. japonica. The newly obtained genetic data are important for further studies of DNA-based taxonomy, population genetics and phylogenetics of the Cosmocercoidea.

A new species of Cosmocercoides (Nematoda; Cosmocercidae) and other helminths in Leptodactylus latrans (Anura; Leptodactylidae) from Argentina.

Cosmocercoides latrans n. sp. (Cosmocercidae) from the small intestine of Leptodactylus latrans (Anura: Leptodactylidae) from Northeastern Province of Buenos Aires, Argentina is described. The new species can be distinguished from their congeners by a combination of the characters, among which stands out the number of rosette papillae, the lack of gubernaculum and the presence of lateral alae in both sexes. There are over 20 species in the genus Cosmocercoides, and Cosmocercoides latrans n. sp. represents the third species from the Neotropical realm and the second for Argentina. Additionally, seven previously known taxa are reported; Pseudoacanthocephalus cf. lutzi, Catadiscus uruguayensis, Rauschiella palmipedis, Aplectana hylambatis, Cosmocerca parva, Schrankiana sp. and Rhabdias elegans; providing literature records and information on distribution and host-parasite relationships.

A new Cosmocercoides species (Ascaridida: Cosmocercidae), C. wuyiensis n. sp., from the Asiatic frog Amolops wuyiensis (Amphibia: Anura).

We identified and characterized a new cosmocercid nematode species, Cosmocercoides wuyiensis n. sp., through microscopic examination and sequencing of the partial small ribosomal RNA gene (18S rDNA), internal transcribed spacer (ITS) and mitochondrial cytochrome c oxidase subunit 1 (COI) genes. The new species was isolated from the intestine of the Asiatic frog Amolops wuyiensis Liu and Hu, 1975 captured from four localities of the Anhui province in south-east China. Among the 25 recorded species of the Cosmocercoides genus, the morphology of C. wuyiensis n. sp. is closest to that of C. kiliwai and C. malayensis, which were isolated from various Mexican frog and Malaysian lizard species, respectively. However, C. wuyiensis n. sp. displayed several distinguishing features, such as small size of the male body, two spicules of unequal lengths in the male, small gubernaculum, pre-, ad- and post-cloacal caudal rosette papillae in the ratio of 18-24:2:6 and simple papillae in the ratio of 14:multiple:4, circle and number of punctation in each rosette at 1:11-16, sharply conical tail-end and the presence of lateral alae and somatic papillae in both sexes. BLAST and the phylogenetic analyses of the 18S rDNA and ITS sequences indicated that C. wuyiensis n. sp. belonged to the genus Cosmocercoides, while that of the COI gene sequence of C. wuyiensis n. sp. showed 16.36% nucleotide divergence with C. pulcher and 47.99% nucleotide divergence with C. qingtianensis. The morphological and molecular characterization of C. wuyiensis n. sp. provides new taxonomic data for this genus.

Molecular identification of potential intermediate hosts of Aulonocephalus pennula from the order Orthoptera.

Aulonocephalus pennula is a heteroxenous nematode that commonly infects a declining game bird, the northern bobwhite quail (Colinus virginianus). There is a lack of information on the life cycle of A. pennula and the potential effects of infection on bobwhites. In order to better understand the life cycle of this parasite, various species from the order Orthoptera were collected from a field site in Mitchell County, Texas. Using polymerase chain reaction (PCR), nine potential intermediate hosts were identified from the 35 orthopteran species collected. Later, ten live specimens were collected to identify larvae within the potential intermediate hosts. Larvae were present in three of these and were sent for sequencing. Similarly, the presence of larvae was confirmed from extra tissues of samples identified as positive with PCR. This was the first study to document potential intermediate hosts, but future studies are needed to confirm that these species are capable of transmitting infection to bobwhite. However, this study demonstrates that PCR has increased sensitivity and may be a valuable tool when determining intermediate hosts.