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.

Baylisascaris procyonis Roundworm Infection in Child with Autism Spectrum Disorder, Washington, USA, 2022.

We describe a case of Baylisascaris procyonis roundworm infection in a child in Washington, USA, with autism spectrum disorder. Environmental assessment confirmed nearby raccoon habitation and B. procyonis eggs. B. procyonis infections should be considered a potential cause of human eosinophilic meningitis, particularly among young children and persons with developmental delays.

Geographic Distribution of Raccoon Roundworm, Baylisascaris procyonis, Germany and Luxembourg.

Infestation with Baylisascaris procyonis, a gastrointestinal nematode of the raccoon, can cause fatal disease in humans. We found that the parasite is widespread in central Germany and can pose a public health risk. The spread of B. procyonis roundworms into nematode-free raccoon populations needs to be monitored.

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 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.

Morphological and molecular evidence for a new species of the genus Cosmocercoides Wilkie, 1930 (Ascaridida: Cosmocercidae) from the Asiatic toad Bufo gargarizans Cantor (Amphibia: Anura).

A new cosmocercid species, Cosmocercoides qingtianensis sp. n., collected from the intestine of the Asiatic toad Bufo gargarizans Cantor (Amphibia: Anura) is described using integrated approaches, including light and scanning electron microscopy, and sequencing and analyzing the ribosomal [small ribosomal DNA (18S) and internal transcribed spacer (ITS)] and mitochondrial [cytochrome c oxidase subunit 1 (cox1)] target regions, respectively. The new species can be distinguished from its congeners by the combination of the following morphological characters, including the large body size, the presence of lateral alae and somatic papillae in both sexes, the length of spicules, the particular morphology and length of gubernaculum, the number, arrangement and morphology of caudal rosettes, the presence of large medioventral precloacal papilla and the long tail. Our molecular analysis revealed the level of intraspecific genetic variation of C. qingtianensis sp. n. distinctly lower than that of the interspecific genetic variation in the ITS and cox1 regions. However, there are some overlaps in the range of intra- and interspecific 18S sequence divergence between the new species and some closely related species. The results of molecular analysis supported the validity of the new species based on the morphological observations. The 18S, ITS, and cox1 regions of C. pulcher collected from Bufo japonicus formosus in Japan were also sequenced and analyzed. The results showed a low level of intraspecific genetic variation in 18S and ITS regions (0-0.12% and 0-0.23% nucleotide differences, respectively), but a relatively high level of intraspecific genetic variation in cox1 region (0.78-4.69% nucleotide differences). In addition, it seems more powerful and practical to use the cox1 region as a genetic marker for the accurate identification and differentiation of species of Cosmocercoides than the 18S and ITS regions, especially for the closely related species.

Morphological and molecular characterization of Megalobatrachonema hainanensis sp. nov. (Nematoda: Ascaridida), with phylogenetic position of Megalobatrachonema in Cosmocercoidea.

The genus Megalobatrachonema is a rare group of nematode parasites within Ascaridida. The systematic status of Megalobatrachonema in the superfamily Cosmocercoidea (Ascaridida) has long been controversial. The relationship of Megalobatrachonema and Chabaudgolvania remains unsolved. In the present study, a new species of Megalobatrachonema, M. hainanensis sp. nov., was described based on specimens collected in Amolops hainanensis (Boulenger) and Hylarana spinulosa (Smith) (Amphibia: Anura) from Hainan Island, China. The large ribosomal DNA (28S) and internal transcribed spacer (ITS1-5.8S-ITS2) were also sequenced for molecular identification and phylogenetic study. Phylogenetic analyses using maximum likelihood (ML) inference and Bayesian inference (BI) based on 28S and ITS1 sequence data, respectively, supported that Megalobatrachonema is a member of the family Kathlaniidae. In addition, the genetic comparison and phylogenetic results based on ITS1 sequence data also supported that the genus Chabaudgolvania should be considered as a synonym of Megalobatrachonema.

Morphology and molecular characterization hold hands: clarifying the taxonomy of Hysterothylacium (Nematoda: Anisakidae) larval forms.

During the last decade, many morphotypes of larval forms of Hysterothylacium have been proposed; however, their boundaries are taxonomically unclear. We compare newly collected material with data available for Hysterothylacium larvae, based on genetics and morphology, in order to evaluate their relationships and simplify their classification. The previously described Hysterothylacium larval type IV, type V, and type X were found in several marine fishes from Brazil. A new Hysterothylacium larval type XV, parasitic in Elops saurus from State of Sergipe, north-eastern Brazil, was proposed based on its genetic singularity and the presence of numerous spines throughout the body. Based upon phylogenetic reconstruction using the ribosomal DNA (rDNA) ITS2 sequences along with morphological traits, the following third stage larvae were considered valid (synonyms are inside parenthesis) type III (= types VII and VIII), type V (= types VI, XII, and XIII), and type X (= type XIV). Despite the lack of molecular data, Hysterothylacium larval type II is considered as synonym of type I because of their identical morphology. The morphometry of Hysterothylacium third stage larvae proved to be weak as a diagnostic tool due to its high rates of variation. Phylogenetic reconstruction using the concatenated sequences of rDNA ITS1-5.8S-ITS2 and the mitochondrion DNA (mtDNA) cox2 revealed that the sampled Hysterothylacium larval types V may represent three different species and H. deardorffovestreetorum may represent two different taxa. Furthermore, H. deardorffovestreetorum is considered species inquirenda due to its problematic description and diagnosis which are based only on larvae. A key to Hysterothylacium larvae from marine fishes is provided.

Detection of parasitic forms in feces of Chelonoidis carbonarius (Linnaeus, 1766) from Brazilian Cerrado.

Chelonoidis carbonarius is host to a great diversity of endoparasites. This research aimed to identify and taxonomically classify the parasitic forms of endoparasites found in the feces of C. carbonarius using coproparasitological methods. In total, 38 fecal samples were collected from animals captured in the municipalities of Cuiabá (n = 23) and Jataí (n = 15) in the Brazilian Ceraado. Fecal samples were submitted to at least one of the following techniques: Willis-Mollay flotation technique, zinc sulfate centrifugal flotation technique, and spontaneous sedimentation of Hoffman, and Pons, and Janer. Overall, 86.85% (33/38) of animals were positive in at least one of the diagnostic techniques. Of the 38 samples used, 50% of the animals were captive and 50% of the animals were free-living. Cysts, oocysts, and parasitic eggs were identified, as follows: Chapiniella spp. (65.78%; n = 24), Sauricola spp. (31.57%; n = 12), Ascaroidea (15.78%; n = 6), Oxyuridae (2.63%; n = 1), Trematoda (2.63%; n = 1), Acanthocephala (2.63%; n = 1), Balantidium spp. (39.47%; n = 15), Eimeria spp. (21.05%; n = 8), and Nyctotherus spp. (2.63%; n = 1). The study showed that both captive and free-living C. carbonarius are often parasitized by different endoparasite species, with nematode eggs being the most common.

Morphology, genetic characterization and molecular phylogeny of the poorly known nematode parasite Cissophyllus leytensis Tubangui & Villaamil, 1933 (Nematoda: Ascaridida) from the Philippine sailfin lizard Hydrosaurus pustulatus (Eschscholtz, 1829) (Reptilia: Squamata).

BACKGROUND: The genus Cissophyllus (Cosmocercoidea: Kathlaniidae) is a rare group of nematodes parasitic in turtles and lizards. To date, only four species have been reported in Asia and North America. However, most of them are inadequately described. The species Cissophyllus leytensis has never been reported since it was originally described by Tubangui and Villaamil in 1933 from the Philippine sailfin lizard Hydrosaurus pustulatus (Eschscholtz, 1829) (Reptilia: Squamata). Furthermore, the systematic status of Cissophyllus/Cissophyllinae in the family Kathlaniidae of the superfamily Cosmocercoidea remains under debate. METHODS: The detailed morphology of C. leytensis was studied using light microscopy (LM) and, for the first time, scanning electron microscopy (SEM), based on newly collected specimens from the type host H. pustulatus. Six different genetic markers, including nuclear sequences [small ribosomal subunit (18S), internal transcribed spacer (ITS) and large ribosomal subunit (28S)], plus mitochondrial genes [cytochrome c oxidase subunit 1 (cox1), cytochrome c oxidase subunit 2 (cox2) and 12S small subunit ribosomal RNA gene] of C. leytensis were sequenced. Additionally, in order to test the validity of the subfamily Cissophyllinae and clarify the phylogenetic relationships of Cissophyllus and the other genera in the family Kathlaniidae, phylogenetic analyses based on 18S + 28S and ITS sequence data were performed using maximum likelihood (ML) and Bayesian inference (BI) analyses, respectively. RESULTS: Our observations using LM and SEM revealed some previously unreported morphological features, necessitating the redescription of this poorly known species. The presence of remarkable morphological variation in the isthmus and the position of excretory pore among different individuals was found. Molecular analysis showed no intraspecific nucleotide divergence detected in the 18S, ITS, 28S, cox2 and 12S regions among different individuals of C. leytensis, but a low level of intraspecific genetic variation was found in the cox1 (0.52%). Our phylogenetic results showed the representatives of the Cosmocercoidea divided into four large clades (Cosmocerca + Aplectana + Cosmocercoides representing the family Cosmocercidae, Cruzia representing the subfamily Cruzinae of Kathlaniidae, Falcaustra + Cissophyllus + Megalobatrachonema representing the subfamily Kathlaniinae of Kathlaniidae, and Orientatractis + Rondonia representing the family Atractidae). The genus Cissophyllus clustered together with the genus Megalobatrachonema in both the ML and BI trees using ITS sequence data, but displayed a sister relationship to the genus Falcaustra in the ML tree and to the genera Falcaustra + Megalobatrachonema in the BI tree using 18S + 28S sequence data. CONCLUSIONS: Molecular phylogenetic results further confirmed that the family Kathlaniidae is not a monophyletic group. The subfamily Cruziinae should be moved from the hitherto-defined family Kathlaniidae and elevated as a separate family Cruziidae. The present phylogenetic results also negated the validity of the subfamily Cissophyllinae and supported the genus Cissophyllus assigned in the subfamily Kathlaniinae. Molecular analysis indicated that the morphological variation in the isthmus and position of excretory pore among different individuals should be considered as intraspecific variation. Moreover, some characters important for the specific diagnosis of C. leytensis are reported for the first time: the number of acuminate denticles (lamellae) on each lip, the chitinized pharynx with three flabellate pharyngeal plates, the presence of single medioventral precloacal papilla and the detailed morphology of caudal papillae. The present study is only the second record of C. leytensis.

Molecular phylogeny of Megalobatrachonema (Nematoda: Ascaridida), with description of a new species based on morphological and molecular evidence.

Species of MegalobatrachonemaYamaguti, 1941 (Ascaridida: Cosmocercoidea) are important nematode parasites in amphibians and reptiles. However, the phylogenetic relationship of its included two subgenera Megalobatrachonema and Chabaudgolvania remains unclear. In the present study, a new species of Megalobatrachonema, M. (Chabaudgolvania) wangi sp. nov., was described based on the specimens collected from the lesser spiny frog Quasipaa exilispinosa (Liu & Hu) (Amphibia: Anura) in China. The ribosomal [large ribosomal DNA (28S) and internal transcribed spacer (ITS1-5.8S-ITS2)] and mitochondrial [12S small subunit ribosomal DNA and cytochrome c oxidase subunit 1 (cox1)] target regions of the new species and M. (Chabaudgolvania) terdentatum, together with the 12S region of M. (Megalobatrachonema) hainanensis, were amplified and sequenced for molecular identification and phylogeny. Moreover, in order to clarify the systematic position of the new species and the phylogenetic relationship of the two subgenera Megalobatrachonema and Chabaudgolvania, phylogenetic analyses based on 28S + ITS1-5.8S-ITS2 + 12S sequence data were performed using maximum likelihood (ML) inference and Bayesian inference (BI). The molecular phylogenetic results conflicted with the current classification and challenged the validity of the subgenus Chabaudgolvania, that should be a synonym of the subgenus Megalobatrachonema. The presence or absence of valves in the oesophageal bulb as a key criterion for delimitation of the two subgenera Megalobatrachonema and Chabaudgolvania seems to be unreliable.

A new species of Cosmocerca (Nematoda, Ascaridomorpha) from the marine toad Rhinella marina (Linnaeus) (Anura, Bufonidae) in Australia.

The marine toad Rhinella marina (Linnaeus) (Anura, Bufonidae) is a notorious, exotic amphibian species in Australia. However, our present knowledge of the composition of the nematode fauna of R. marina is still not complete. In the present study, a new cosmocercid nematode, Cosmocerca multipapillata sp. nov., was described using both light and scanning electron microscopy, based on specimens collected from R. marina in Australia. Cosmocerca multipapillata sp. nov. can be easily distinguished from its congeners by the body size, the presence of lateral alae and well sclerotized gubernaculum, the number and arrangement of plectanes and rosettes and the length of spicules, oesophagus and tail.

Redescription of Porrocaecum semiteres (Zeder, 1800) (Nematoda: Ascaridida) from the Song Thrush Turdus philomelos (Passeriformes: Turdidae).

INTRODUCTION: Porrocaecum semiteres (Zeder, 1800) (Ascaridida: Ascaridoidea) occurs frequently in various birds in Europe. However, the knowledge of the morphology of this species remains insufficient. METHODS: Light and scanning electron microscopy were used to study the detailed morphology of adults of P. semiteres based on newly collected specimens from Turdus philomelos in the Czech Republic. RESULTS: Some previously unreported morphological features were revealed, including the presence of a medio-apical notch, 70-80 small conical denticles and one pair of submedial pores on each lip, the absence of caudal alae, the presence of four pairs of postcloacal papillae, and a single, medio-ventral precloacal papilla.

Morphological variability, ultrastructure and molecular characterisation of Hysterothylacium reliquens (Norris & Overstreet, 1975) (Nematoda: Raphidascarididae) from the oriental sole Brachirus orientalis (Bloch & Schneider) (Pleuronectiformes: Soleidae).

Hysterothylacium reliquens (Norris & Overstreet, 1975), a common ascaridoid nematode parasitic in the marine fishes in the Indian and Atlantic Oceans, has been reported to exhibit remarkable morphological variability in the lengths of body, oesophagus and spicules, and the number and arrangement of postcloacal papillae. In order to determine whether H. reliquens with a broad range of morphological variability in the above-mentioned respects, represents a complex of sibling species or a single species, and to evaluate if the specimens of H. reliquens collected from the Indian and Atlantic Oceans are the same species, numbers of H. reliquens collected from the oriental sole Brachirus orientalis (Bloch & Schneider) (Pleuronectiformes: Soleidae) in the Arabian Gulf (off Iraq) were characterised using molecular approaches by sequencing and analysing the ribosomal [large ribosomal DNA (28S) and internal transcribed spacer (ITS)] and mitochondrial [cytochrome c oxidase subunit 2 (cox2)] target regions, respectively. The results of molecular analysis strongly supported that: (i) the broad ranges of morphological variability in the above-mentioned respects in the newly collected specimens of H. reliquens, should be considered as intraspecific variation and the nematode material collected from B. orientalis in the Arabian Gulf represented a single species; (ii) the specimens of H. reliquens collected from the Indian and Atlantic Oceans belong to the same species, but they possibly represent different geographical populations. Moreover, the detailed morphology of the labial papillae, lateral alae, excretory pore, distal end of spicules, midventral precloacal papilla, phasmids and caudal papillae was revealed by SEM for the first time. The molecular and morphological data of H. reliquens obtained herein contribute to a more accurate diagnosis of this little known ascaridoid nematode.

Parasitism of molluscs by nematodes: types of associations and evolutionary trends.

Although there are no confirmed fossil records of mollusc parasitic nematodes, diverse associations of more than 108 described nematode species with slugs and snails provide a fertile ground for speculation of how mollusc parasitism evolved in nematodes. Current phylogenic resolution suggests that molluscs have been independently acquired as hosts on a number of occasions. However, molluscs are significant as hosts for only two major groups of nematodes: as intermediate hosts for metastrongyloids and as definitive hosts for a number of rhabditids. Of the 61 species of nematodes known to use molluscs as intermediate hosts, 49 belong to Metastrongyloidea (Order Strongylida); of the 47 species of nematodes that use molluscs as definitive hosts, 33 belong to the Order Rhabditida. Recent phylogenetic hypotheses have been unable to resolve whether metastrongyloids are sister taxa to those rhabditids that use molluscs as definitive hosts. Although most rhabditid nematodes have been reported not to kill their mollusc hosts prior to their reproduction, some species are pathogenic. In fact, infective juveniles of Phasmarhabditis hermaphrodita vector a lethal bacterium into the slug host in which they reproduce. This life cycle is remarkably similar to the entomopathogenic nematodes in the families Steinernematidae and Heterorhabditidae. Also, the discoveries of Alloionema and Pellioditis in slugs are interesting, as these species have been speculated to represent the ancestral forms of the entomopathogenic nematodes. Development of the infective stage appears to be an important step toward the acquisition of molluscs as definitive hosts, and the association with specific bacteria may have arisen in conjunction with the evolution of necromeny.

Primeros registros de Hysterothylacium pelagicum (Anisakidae) y Toxocara alienata (Toxocaridae) en el Perú - Fe de errata

Error en el artículo Primeros registros de Hysterothylacium pelagicum (Anisakidae) y Toxocara alienata (Toxocaridae) en el Perú publicado en la Revista Peruana de Biologia (2014) 21 (2): 171-174. DOI: http://dx.doi.org/10.15381/rpb.v21i2.9820. El error esta referido a la Tabla 1. Algunas medidas de Hysterothylacium pelagicum no han sido expesadas correctamente. Debe considerarse la tabla adjunta.

An error is in the article First records of Hysterothylacium pelagicum (Anisakidae) and Toxocara alienata (Toxocaridae) from Peru published in Revista Peruana de Biologia (2014) 21 (2): 171-174. DOI: http://dx.doi.org/10.15381/rpb.v21i2.9820. The error is referred to Table 1. Some measures of Hysterothylacium pelagicum are not reported correctly. Here, present the table that should be considered.

Primeros registros de Hysterothylacium pelagicum (Anisakidae) y Toxocara alienata (Toxocaridae) en el Perú

Dos nematodos ascarididos son registrados para el Perú: Hysterothylacium pelagicum parasitando al pez perico o dorado (Coryphaena hippurus) colectado en un mercado de Lima y a Toxocara alienata encontrado en un pecarí de collar (Pecari tajacu) procedente de Iquitos. Así mismo, se realiza una descripción morfológica de los nematodos mencionados.

Two ascaridoid nematodes are recorded by Peru: Hysterothylacium pelagicum parasitizing to common dolphinfish (Coryphaena hippurus) from a market of Lima and Toxocara alienata in a collared peccary (Pecari tajacu) from Iquitos. Likewise, it is made a morphological description of the nematodes.