Potential anthelmintic effect of chitosan on Syphacia muris infecting Wistar rats: biochemical, immunological, and histopathological studies.

Natural products extracted from animal sources have many biological activities, such as chitosan, which is being researched for its medicinal or therapeutic potential. Syphacia muris is the most well-known intestinal nematode, infecting laboratory rats and influencing their immune systems. In this study, we looked at the anthelminthic activity of chitosan particles against S. muris infection using biochemical, immunological, and histopathological methods. Chitosan particles were characterized using Fourier-transform infrared spectroscopy (FTIR). Rats were separated into four groups, each consisting of seven individuals (n = 7). The first group was the control (non-infected), the second group was infected, and both groups received 0.5 ml of 1% glacial acetic acid orally. The third group was the infected group (treated), and the fourth group (normal) received 0.5 ml of 30 mg/kg/day chitosan dissolved in 1% glacial acetic acid for 14 days using gavage. Liver and kidney parameters, oxidative stress markers, serum levels of cytokines (IFN-γ, IL-5, IL-13, IL-33, and IL-10), as well as immunoglobulins (total IgE and IgG), were assessed. Histological examinations of host tissues (intestine, liver, kidney, and spleen) were also performed. Following chitosan treatment, a significant decrease in worm count (P < 0.05) was indicated; this was associated with an enhancement of biochemical and oxidative stress biomarkers, which were altered due to infection. Moreover, immunological analysis revealed a significant drop in INF-γ, IL-5, IL-13, and IL-33 levels and total immunoglobulins (IgE and IgG) as well as an improvement in rat tissues. Conclusively, this study showed the anthelminthic effect of chitosan against S. muris infection.

MOLECULAR PHYLOGENETIC ANALYSIS OF GYRINICOLA BATRACHIENSIS (NEMATODA: OXYUROIDEA) LENDS SUPPORT TO THE MONOPHYLY OF THE GENUS AND THE RESURRECTION OF GYRINICOLINAE.

Gyrinicola Yamaguti, 1938, includes 6 species of oxyurid found within the intestinal tract of numerous, larval, anuran species in Europe, Asia, South America, and North America. The systematic placement and hierarchical treatment of the genus has shifted at least 5 times since its discovery; the group was first considered as its own family (Gyrinicolidae), then treated as a subfamily (Gyrinicolinae) of Cosmocercidae, then as a member of the Pharyngodonidae, followed by movement back to the Cosmocercidae, and finally a recent proposal suggested the resurrection of the Gyrinicolidae. Species in the genus vary widely in the morphology of the uterine tract, a characteristic often used to indicate membership in the genus, as it is tied to the reproductive mode. However, until recently very few genetic data were available to aid in the placement of this unique group of worms, and before this study to the best of our knowledge none existed for the North American species. To examine the monophyly and placement of the Gyrinicola we sampled populations of Gyrinicola batrachiensis across North America and screened them for genetic diversity using nuclear markers 18S and 28S. Phylogenies suggest at least 3 clades exist among the nematodes from North America and that these clades, alongside Gyrinicola japonica, form a well-supported group within Oxyuroidea. Further representation of Pharyngodonidae from other vertebrate classes may help clarify the relationship of this historical grouping to other members of the Oxyuroidea.

Molecular data aids pinworm diagnosis in night monkeys (Aotus spp., Primates: Aotidae) with the resurrection of a Trypanoxyuris species (Nematoda: Oxyuridae).

Neotropical primates (Platyrrhines) are commonly parasitized by pinworm nematodes of the genus Trypanoxyuris Vevers, 1923. The taxonomic identity of Trypanoxyuris sampled in night monkeys (Aotus Iliger) has been rather controversial. Two species have been described, namely T. microon (Linstow, 1907) and T. interlabiata (Sandosham, 1950). The latter was synonymized with T. microon considering that the observed morphological differences corresponded to different developmental stages of the nematode rather than to differences between both species. Here, we used an integrative taxonomy approach, based on morphological and molecular data along with host identity, in order to assess the validity of both species. Our results evidenced that these different morphotypes correspond to different and reciprocally monophyletic groups; thus, we propose the resurrection of T. interlabiata. We redescribe both pinworm species using specimens sampled in Aotus monkeys from Colombia and discuss the advantages of combining molecular and morphological data to uncover pinworm diversity, and to understand the potential forces determining the diversification process in pinworms from platyrrhine primates.

The complete mitochondrial genome of Wellcomia compar (Spirurina: Oxyuridae) and its genome characterization and phylogenetic analysis.

Wellcomia compar (Spirurina: Oxyuridae) is a pinworm that infects wild and captive porcupines. Despite clear records of its morphological structure, its genetics, systematics, and biology are poorly understood. This study aimed to determine the complete mitochondrial (mt) genome of W. compar and reconstruct its phylogenetic relationship with other nematodes. We sequenced the complete mt genome of W. comparand conducted phylogenetic analyses using concatenated coding sequences of 12 protein-coding genes (PCGs) by maximum likelihood and Bayesian inference. The complete mt genome is 14,373 bp in size and comprises 36 genes, including 12 protein-coding, two rRNA and 22 tRNA genes. Apart from 28 intergenic regions, one non-coding region and one overlapping region also occur. A comparison of the gene arrangements of Oxyuridomorpha revealed relatively similar features in W. compar and Wellcomia siamensis. Phylogenetic analysis also showed that W. compar and W. siamensis formed a sister group. In Oxyuridomorpha the genetic distance between W. compar and W. siamensis was 0.0805. This study reports, for the first time, the complete W. compar mt genome sequence obtained from Chinese porcupines. It provides genetic markers for investigating the taxonomy, population genetics, and phylogenetics of pinworms from different hosts and has implications for the diagnosis, prevention, and control of parasitic diseases in porcupines and other animals.

Ozolaimus megatyphlon and Ozolaimus cirratus parasitizing the Iguana iguana (Linnaeus, 1758) from Marajó Island, Pará, Brasil: new occurrence and morphological redescription.

This study aimed to redescribe two species of Ozolaimus, parasites of free-living green iguanas native to Marajó Island. The gastrointestinal system of four iguana specimens was evaluated for the presence of helminths. Altogether, 12,028 nematodes were found, with a prevalence of 100%, an infection range of 780 to 7,736 nematodes, an infection intensity of 3.007, and a mean abundance of 3,007. Light microscopy and scanning electron microscopy were used to determine the species of nematodes found. The cecum was the site of infection that had the highest parasitic load. Morphologically, the nematodes were compatible with the genus Ozolaimus Dujardin, 1844, with the species Ozolaimus megatyphlon (Rudolphi, 1819) Dujardin, 1845, and Ozolaimus cirratus Linstow, 1906. Scanning electron microscopy showed the presence of small structures (serrated in Ozolaimus cirratus and rounded in Ozolaimus megatyphlon) located below the esophageal leaves. We also evidenced the phasmids in both species; this is the first record of these structures in nematodes of the genus Ozolaimus. In addition, this work expands the records on the geographic distribution of these parasites.

Diagnosis of endoparasite species and subtypes of Cryptosporidium spp. with one health importance, in feces from captive snakes in Rio de Janeiro, Brazil.

In captivity, snakes may present chronic infections with high mortality, such as those caused by Cryptosporidium serpentis, or they may be pseudoparasitized by species that present zoonotic potential. Thus, the aim of this study was to evaluate the frequency of helminths and protozoa in the feces of captive snakes, characterize the species and subtypes of Cryptosporidium spp. and correlate the parasites detected with other information obtained from these animals. Feces were collected from 189 snakes kept at the Vital Brazil Institute, Rio de Janeiro, including samples from Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops atrox, Bothrops leucurus, Crotalus durissus and Lachesis muta. All the samples were subjected to microscopy techniques and the polymerase chain reaction (PCR) in association with sequencing, to identify Cryptosporidium spp.. Forms of parasites infecting the snakes were identified through microscopy in 50.8% of the samples. Helminths were detected more often than protozoa in the feces of these animals, mainly comprising eggs resembling Kalicephalus sp. and oocysts of Eimeria sp.. Pseudoparasites such as Syphacia sp., Aspiculuris sp. and Hymenolepis nana were also detected. Through correlating the results obtained from parasitological staining techniques and PCR, the total frequency of Cryptosporidium sp. was found to be 19%. The species C. tyzzeri and C. parvum were identified. Characterization using the target gp60 showed subtypes with high potential for zoonotic transmission, especially IIaA15G2R1 and IIaA14G2R1 of C. parvum and IXbA8 of C. tyzzeri. This study highlighted the need for more intensive health management in the Institute's serpentarium and, especially, in its bioterium where rodents are reared as a food source for these snakes.

Skrjabinodon castillensis n. sp. (Nematoda:Pharyngodonidae) from the Homonota horrida and H.darwinii (Squamata: Phyllodactylidae) from Argentina and key for the Neotropical species of the genus Skrjabinodon.

Skrjabinodon castillensis n. sp. is described and illustrated here, based on specimens found in the large intestines of Homonota horrida (province San Juan) and Homonota darwinii (province Neuquén) (Squamata: Phyllodactylidae) from Argentina. The new species is assigned to Skrjabinodon based lateral alae present in males, absent in females. Lateral alae beginning midway between lips and nerve ring and ending just posterior to first pair of caudal papillae. Females with vulva near esophageal bulb. In males, caudal alae absent, paired caudal papillae present. Skrjabinodon castillensis n. sp. represents the 9th species from the Neotropical realm. The new species differs from all other species assigned to Skrjabinodon by morphology of tail filament and number of tail filament spines. Skrjabinodon castillensis n. sp. is the only species of this genus known from Argentina. A key to the species of Skrjabinodon in the Neotropical realm is provided.

New host and locality records of helminths' infection of seven lizards from Morocco.

Morocco has a great diversity of reptiles; more than 26% of the herpetofauna species are considered to be endemic. Nevertheless, there is little information available on helminth parasites of Moroccan lizards. The purpose of this article is to establish a helminth list using non-invasive methods for natural populations of seven lizard species: Acanthodactylus maculatus, Chalcides mionecton, Chalcides montanus, Chalcides polylepis, Quedenfeldtia moerens, Quedenfeldtia trachyblepharus, and Tarentola mauritanica. For each species, prevalence and intensity of infection were given as well as their relationship to some determinant parameters (sex, age class, size, altitude, and seasons). Four species of nematodes, Parapharyngodon micipsae, Pharyngodon mamillatus, Spauligodon auziensis, Thelandros alatus, and unassigned Spauligodon were found. Mean helminth intensity for the seven lizard species was 7.3 ± 16.2 SD (range = 1-92 in infected lizards), with total infection prevalence of 5.6%. Mixed infections were observed in Chalcides montanus and Chalcides polylepis co-infected by Pharyngodon mamillatus and Spauligodon sp. Furthermore, the intensity of infection by nematodes were significantly different among the seven studied species. The nematode infection in Chalcides mionecton were related to sex, host size, and altitude; in Chalcides polylepis to the host age; in Quedenfeldtia trachyblepharus to elevation, age, and host size; and in Tarentola Mauritanica to age, body size, seasons, and altitude. In conclusion, our study provides data about nine new host and locality records and the effect of some determinant factors on host parasitism.

New species of Parapharyngodon (Nematoda: Pharyngodonidae) parasite of Osteocephalus taurinus (Anura: Hylidae) from Northern Brazilian Amazon Region.

Parapharyngodon spp. are nematodes parasites of amphibian and reptiles worldwide distributed. Of 55 valid species, only 11 are known for the Neotropical realm, and six were found parasitizing hylid frogs in the world. In the present work, we describe a new species of Parapharyngodon from the large intestine of Osteocephalus taurinus from Reserva de Desenvolvimento Mamirauá, Amazonas, Brazil, using light and scanning electron microscopy. Parapharyngodon curupira n. sp. is allocated to the group of species with three pairs of cloacal papillae. However, it shows a set of different morphological characters compared to its congeners. In the new species, males have gubernaculum and smooth cloacal lip, females have pre-bulbar ovaries, and we also observed some morphometric differences. Among Parapharyngodon spp. from the Neotropical realm, the new taxon resembles P. alverangai and P. politoedi. The last species was described from the same host and biome (Amazon); however, they can be easily distinguished from new species by the presence of gubernaculum and v-shape structure at the lower caudal lip. Thus, we characterize and propose a new species of Parapharyngodon from the western amazon.

Pharyngodon mamillatus (Nematoda: Pharyngodonidae) in the ocellated skink Chalcides ocellatus (Squamata: Scincidae) from Egypt: Light and scanning electron microscopic studies.

Twenty-one Pharyngodon mamillatus were recovered from the large intestine of 4 out of 12 ocellated skink Chalcides ocellatus collected from the South Sinai desert, Egypt. The prevalence of infection was 33.33% (intensity 2-4). The recovered pharyngodonid was re-described based on light and scanning electron microscopy. According to the morphology, it was characterized by regular transverse annulations extending from the posterior margin of the lips to the level of the anus, triangular mouth opening was surrounded by three large bilobed lips possessing three papillae, oval-shaped excretory pore, slit-like vulva opening, and slit-like anal ventral opening at the posterior end which terminates with a tail filament. Here, the morphometric measurements of adult female P. mamillatus were given in this study. It has no intraspecific morphological variations from the previously described P. mamillatus, but it appeared with small range on biometric features. These variations may be accounted by host/environment attributes since they exhibit wide host and geographical spectra. The present study was applied for detailed morphological knowledge of the recovered P. mamillatus for strengthening their taxonomic validity. HIGHLIGHTS: The Pharyngodon mamillatus naturally parasite of the intestinal region of Chalcides ocellatus. The present study clarifies the morphological key for the description of P. mamillatus.

Parasitic nematodes of the genus Syphacia Seurat, 1916 infecting Cricetidae in the British Isles: the enigmatic status of Syphacia nigeriana.

Oxyurid nematodes (Syphacia spp.) from bank (Myodes glareolus) and field/common (Microtus spp.) voles, from disparate geographical sites in the British Isles, were examined morphologically and genetically. The genetic signatures of 118 new isolates are provided, based primarily on the rDNA internal transcribed spacers (ITS1-5.8S-ITS2) region and for representative isolates also on the small subunit 18S rDNA region and cytochrome c oxidase subunit 1 (cox-1) gene locus. Genetic data on worms recovered from Microtus spp. from the European mainland and from other rodent genera from the Palaearctic, North America and West Africa are also included. We test historical hypotheses indicating that S. nigeriana is a generalist species, infecting a range of different rodent genera. Our results establish that S. nigeriana is a parasite of both bank and field voles in the British Isles. An identical genotype was also recorded from Hubert's multimammate mouse (Mastomys huberti) from Senegal, but Mastomys spp. from West Africa were additionally parasitized by a related, although genetically distinct Syphacia species. We found no evidence for S. petrusewiczi in voles from the British Isles but isolates from Russia and North America were genetically distinct and formed their own separate deep branch in maximum likelihood molecular phylogenetic trees.

Co-structure analysis and genetic associations reveal insights into pinworms (Trypanoxyuris) and primates (Alouatta palliata) microevolutionary dynamics.

BACKGROUND: In parasitism arm race processes and red queen dynamics between host and parasites reciprocally mold many aspects of their genetics and evolution. We performed a parallel assessment of population genetics and demography of two species of pinworms with different degrees of host specificity (Trypanoxyuris multilabiatus, species-specific; and T. minutus, genus-specific) and their host, the mantled howler monkey (Alouatta palliata), based on mitochondrial DNA sequences and microsatellite loci (these only for the host). Given that pinworms and primates have a close co-evolutionary history, covariation in several genetic aspects of their populations is expected. RESULTS: Mitochondrial DNA revealed two genetic clusters (West and East) in both pinworm species and howler monkeys, although population structure and genetic differentiation were stronger in the host, while genetic diversity was higher in pinworms than howler populations. Co-divergence tests showed no congruence between host and parasite phylogenies; nonetheless, a significant correlation was found between both pinworms and A. palliata genetic pairwise distances suggesting that the parasites' gene flow is mediated by the host dispersal. Moreover, the parasite most infective and the host most susceptible haplotypes were also the most frequent, whereas the less divergent haplotypes tended to be either more infective (for pinworms) or more susceptible (for howlers). Finally, a positive correlation was found between pairwise p-distance of host haplotypes and that of their associated pinworm haplotypes. CONCLUSION: The genetic configuration of pinworm populations appears to be molded by their own demography and life history traits in conjunction with the biology and evolutionary history of their hosts, including host genetic variation, social interactions, dispersal and biogeography. Similarity in patterns of genetic structure, differentiation and diversity is higher between howler monkeys and T. multilabiatus in comparison with T. minutus, highlighting the role of host-specificity in coevolving processes. Trypanoxyuris minutus exhibits genetic specificity towards the most frequent host haplotype as well as geographic specificity. Results suggest signals of potential local adaptation in pinworms and further support the notion of correlated evolution between pinworms and their primate hosts.

Anti-pinworm activity of novel coumarin-based trisubstituted methanes in Syphacia obvelata-infected mice.

The control of pinworms mainly relies on use of anthelmintic drugs. At present, there exists only few medications against pinworms, and their repeated use pose a serious risk of resistance development. Therefore, new anti-pinworm drugs are required to overcome the risk of resistance. This study reports the anti-pinworm activity of three novel coumarin-based trisubstituted methanes (TRSMs), i.e., 6-Amino-5-((4-hydroxy-2-oxo-2H-chromen-3-yl)(4-fluoro-phenyl)methyl)-1,3-dimethyl-pyrimidine-2,4(1H,3H)-dione (1), 6-Amino-5-((4-hydroxy-2-oxo-2H-chromen-3-yl)(4-chlor-ophenyl)methyl)-1,3-dimethyl-pyrimidine-2,4(1H,3H)-dione (2) and 6-Amino-5-((4-hydroxy-2-oxo-2H-chromen-3-yl)(4-bromophenyl)methyl)-1,3-dimethyl-pyrimidine-2,4(1H,3H)-dione (3) in Syphacia obvelata-infected mice. The oral acute toxicity of compounds was examined using the OECD guidelines. The findings of this study reveal that TRSM analogues 1 and 2, at a single 80 mg/kg dose given for 5 days, can reduce about 90% of pinworm worm burden in mice, compared to 98% worm reduction shown by 20 mg/kg dose of albendazole, the reference drug, on the 12 day of infection. In particular, the fluoro-and bromo-substituents in the phenyl ring of synthesized derivatives greatly influence the efficacy of candidates. The oral acute toxicity of TRSMs was observed to be greater than 2000 mg/kg body weight for mice. Taken together, our study suggests that studied novel coumarin-based trisubstituted methanes could serve as suitable candidates for the development of new anti-pinworm drugs.

Nematodes from the Tasmanian devil (Sarcophilus harrisii (Boitard)), with the description of Sarcophiloxyuris longus n. gen. and n. sp. (Oxyuridae).

The Tasmanian devil (Sarcophilus harrisii (Boitard)) is an endangered carnivorous marsupial, limited to the islands of Tasmania in southern Australia. The parasites of the Tasmanian devil are understudied. This study aimed to increase the knowledge of the nematode fauna of Tasmanian devils. Ten Tasmanian devils were examined for parasites from northern and southern Tasmania. Nematodes that were collected were morphologically characterized as two separate species. Molecular sequencing was undertaken to verify the identity of these species. A new genus and species of oxyurid nematode was collected from a single Tasmanian devil from the northern part of Tasmania. The nematode is differentiated from oxyurids described from other Australian amphibians, reptiles and marsupials by the characters of the male posterior end - that is, in having three pairs of caudal papillae, two pairs peri-cloacal, one large pair post-cloacal, a long tapering tail, a stout spicule and a gubernaculum and accessory piece, as well as its much larger overall size. Molecular sequencing was unsuccessful. The remaining nematodes collected from the Tasmanian devil in this study were all identified as Baylisascaris tasmaniensis Sprent, 1970, through morphology and molecular sequencing. This paper presents the first description of a new genus and species of oxyurid nematode from the Tasmanian devil, Sarcophiloxyuris longus n. gen., n. sp. The need to undertake more sampling of the parasites of endangered hosts, such as the Tasmanian devil, to assist with a better understanding of their conservation management, is discussed.

Parapharyngodon hispidus n. sp. (Nematoda: Pharyngodonidae) in Tropidurus hispidus (Spix) (Squamata: Tropiduridae) from Caatinga Biome of the Vale do São Francisco, state of Pernambuco, Brazil with a key for the Neotropical species of the genus Parapharyngodon Chatterji.

One species of Parapharyngodon Chatterji (Nematoda: Pharyngodonidae), parasitizing the large intestine of Tropidurus hispidus (Spix) (Squamata: Tropiduridae) in Caatinga biome from Brazil is described and illustrated . This new species differs from a part of its congeners by a set of characters in males: the presence of four pairs of caudal papillae, unpaired post cloacal papilla absent, and smooth anterior cloacal lip. Among the Parapharyngodon species with four pairs caudal papillae, unpaired post clocal papilla absent, and smooth cloacal lips, only P. hispidus n. sp. and P. jairaipurii Rizvi Bursey (Oriental realm), features the same characteristics including sharp pointed tip of spicule, stout terminus or spike of tail of females, and punctate ornamentation of the eggshell. However, these two species are different by the size of spicules (spicules of P. jairaipuri are around 55% of the size spicules of P. hispidus n. sp.); and the arrangement of the caudal papillae (two pairs of adcloacal papillae in P. jaraipuri which are absent in P. hispidus n. sp.). Parapharyngodon hispidus n. sp. is the 55th species assigned to the genus and the 11th species of this genus described in Neotropical hosts.

Genetic diversity of Syphacia Seurat, 1916 (Nematoda: Oxyuridae) across the hybrid zone of their rodent hosts in Russia.

The genetic diversity of Syphacia nematodes (intestinal parasites of rodents) was studied in the hybrid zone of two sister species of common voles, Microtus arvalis and Microtus obscurus, in the Oka River valley, east of Moscow. Syphacia nematodes of other rodent species (Microtus rossiaemeridionalis, Alexandromys oeconomus, Sylvaemus uralensis, and Apodemus agrarius) that inhabit the area were also studied. Phylogenetic trees for the studied nematodes were inferred from the analysis of nuclear ITS1+5.8S+ITS2, LSU rDNA, and mitochondrial CO1 gene partial sequences. Syphacia nematodes of the studied area form three well-defined clades in the phylogenetic tree of this genus. Morphological analysis revealed similarities between the obtained sequences with those of known Syphacia species from the GenBank database, which enabled identifying these three clades up to the species level: S. montana, S. agraria, and S. frederici. Russian haplotypes of Syphacia are different from West European and East Asian haplotypes with pronounced genetic distances. A high level of specificity was reported for two of these three species (S. frederici, only in Sylvaemus uralensis; S. agraria, only in Apodemus agrarius). S. montana was found in different species of voles. Remarkably, S. montana specimens from M. arvalis and M. obscurus were genetically uniform, while S. montana, specimens from hybrids between these two species formed a separate clade distant from those originating from non-hybridised hosts.

A new species of Wellcomia (Nematoda: Oxyuridae) in the plains viscacha (Rodentia: Chinchillidae) from Argentina, an emended diagnosis and an update of the genus Wellcomia.

Wellcomia species (Oxyuridae, Syphaciinae) parasitise several families of rodents from Africa, Asia and America with diverse ecological characteristics. A new species of Wellcomia is described based on specimens obtained from the plains viscacha Lagostomus maximus (Chinchillidae) from Argentina. The description includes morphometric and ecological aspects and an emended diagnosis of the genus Wellcomia. In addition, a molecular characterisation (18S rRNA) and an exploratory analysis of the genetic distances of the species included in this genus are provided. The new species, Wellcomia hugoti n. sp., differs from other species of the genus in the morphology of the cephalic plate, the ventral rugose area in males and the vulva in females. The new species showed a low prevalence (7.7%) and a high mean abundance (73.8). The genetic distances detected do not allow a comprehensive assessment of the monophyly of the genus. This is the seventh record of this genus in rodents from the Americas, the second in rodents from Argentina and the first record for the family Chinchillidae.

Standardization of an LNA-based TaqMan assay qPCR analysis for Aspiculuris tetraptera DNA in mouse faeces.

BACKGROUND: Aspiculuris tetraptera, as a parasitic pinworm, is most frequently detected in laboratory mice, and transmission is mediated by the eggs contained in the faeces of infected mice. A highly sensitive and quantitative faeces-based diagnostic tool would be useful for the early detection of A. tetraptera to inhibit the expansion of infection. In this study, we developed a quantitative assay that exhibits high sensitivity in detecting A. tetraptera in faeces using PCR techniques. RESULTS: Endpoint PCR demonstrated the detection of A. tetraptera DNA in 0.5 ng genomic DNA extracted from the faeces of infected mice. To quantitatively detect the small amount of A. tetraptera DNA, locked nucleic acid (LNA)-based primers and LNA-based TaqMan probes were used for the quantitative PCR assay (qPCR). The combination of LNA-based DNA increased detection sensitivity by more than 100-fold compared to using normal oligo DNAs. The copy number of the A. tetraptera DNA detected was positively related to the infected faeces-derived genomic DNA with a simple linearity regression in the range of 20 pg to 15 ng of the genomic DNA. To more conveniently detect infection using faeces, the LNA-based TaqMan assay was applied to the crude fraction of the faeces without DNA purification. An assay using ethanol precipitation of the faeces yielded results consistent with those of direct microscopic observation. CONCLUSION: The LNA-TaqMan assay developed in this study quantitatively detects A. tetraptera infection in mouse faeces.

Batracholandros salamandrae (Oxyuroidea: Pharyngodonidae) in Endemic Salamanders (Amphibia: Plethodontidae) of the Trans-Mexican Volcanic Belt: Host Range Wide Distribution or Cryptic Species Complex?

Salamanders of the tribe Bolitoglossini Hallowell are a highly diversified group of amphibians, and their helminth parasite fauna has been scarcely studied. Some species of plethodontid salamanders distributed along the Trans-Mexican Volcanic Belt, in central Mexico, were sampled, and their helminth parasites were recovered for taxonomic identification. Specimens of a pharyngodonid nematode from 2 species of bolitoglossines of the genus Pseudoeurycea Taylor were morphologically identified as Batracholandros salamandrae (Schad, 1960) Petter and Quentin, 1976. These specimens were studied in further detail through light and scanning electron microscopy and were sequenced for 2 ribosomal genes and 1 mitochondrial gene to test the hypothesis of whether B. salamandrae is a species widely distributed in salamanders across the Nearctic biogeographic region, or if it represents a cryptic species complex. Our molecular results revealed that these specimens consisted of 2 genetic lineages in concordance with host species, although with slight morphological differences among specimens in each of them. A thorough study, including the generation of molecular data from individuals from other areas of North America, and the examination of type specimens, is required to test the reliability of these morphological differences and to corroborate the species identity of the 2 genetic lineages.

Oxyuricassis ekstromi n. sp. (Oxyurida: Pharyngodonidae) from Lasiancistrus saetiger (Siluriform: Loricariidae) from the Eastern Amazon.

Oxyuricassis ekstromi n. sp. is the third species of the genus described from the intestine of Lasiancistrus saetiger Armbruster in Brazil. This species can be easily differentiated from its congeners by the presence of a tooth-like sclerotized piece that occupies half of the stoma and absence of spine projections on the tail of both males and females. Also, males of the new species lack caudal alae present in Oxyuricassis coronatus and Oxyuricassis hexaspinatus. Additionally, we observed a different number and pattern of the distribution of cloacal papilla; males of O. coronatus and O. hexaspinatus have 1 pair of pre-cloacal and 2 pairs of post-cloacal papillae; while the new species has 1 pre-cloacal and 3 post-cloacal pairs of papillae.