GENETIC AND TRAIT VARIABILITY OF GYRINICOLA REVEALS THE EXISTENCE OF AT LEAST FOUR SPECIES WITHIN THE UNITED STATES.

The tadpole-dwelling pinworm, Gyrinicola batrachiensis (Walton, 1929) Adamson, 1981 was recognized as the sole representative of the genus across Canada and the United States. However, evaluation of the morphology of these parasites across their range revealed considerable morphological variability that suggested diagnosable morphotypes. These morphotypes were associated with different species of anurans, several of which occurred in sympatry. Herein we use an extensive geographic sampling across the United States to obtain the morphotypes, screen their genetic diversity, and analyze this information using an integrative approach. We reconstructed their phylogeny using nuclear ribosomal partial genes 18S and 28S, ITS1, 5.8S, and ITS2, as well as 5 mitochondrial genes generated with Next-Generation sequencing technology. This phylogeny reveals 3 well-resolved lineages, which upon the use of a statistical approach (bPTP [Bayesian implementation of the Poisson tree processes]) supports the delimitation of 4 distinct groups equivalent to species. These putative species groups were tested using morphological characteristics paired with a MANOVA and canonical variate analysis. Results suggest that at least 4 species of Gyrinicola are present within North America, resulting in the resurrection of G. armatus (Walton, 1933) and the description of 2 new species.

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.

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.

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.

Morphological description and phylogenetic assessment of 28S rRNA for Thelandros chalcidiae sp. nov. from Chalcides ocellatus.

Thelandros is a genus of oxyurid nematodes which parasitize both omnivorous and herbivorous hosts. Thelandros chalcidiae sp. nov. is a new taxa described from the large intestine of the ocellated skink, Chalcides ocellatus, from the South Sinai Desert in Egypt. The recovered parasite species was examined at both morphological and molecular levels in order to determine the exact taxonomic position within Pharyngodonidae family. The current pharyngodonid species is characterized by a mouth opening bounded by three bilobed lips; male worms are characterized by the presence of two pairs of cloacal papillae (pre- and adcloacal) and one single caudal papillae (postcloacal) and caudal alae in an auricular form and females with post-equatorial vulva, amphidelphic ovary in which anterior ovary extended to level of excretory pore and posterior ovary extended posteriorly up anal opening. The recovered nematodes were compared with other known species from different hosts and it was found to be morphologically different from them. Molecular characterization based on the partial 28S rRNA nuclear ribosomal gene sequence showed sequence identities ≥ 83.15% with taxa under family Pharyngodonidae, 74.84-87.37% with Oxyuridae, 80.54% with Heteroxynematidae, and 75.98-77.72% with Thelastomatidea. Phylogenetic analysis showed that parasite sequence in conjunction with existing data facilitate placement of this species within Oxyurida. The present species is deeply embedded in genus Thelandros with close relationships to previously described Thelandros sp. and T. galloti in same taxon. This study highlighted importance of combining morphological and genetic data with taxonomy in pharyngodonid species.

Morphological redescription and phylogenetic assessment of Spauligodon aspiculus (Oxyuroidea: Pharyngodonidae) infecting the white-spotted gecko, Tarentola annularis (Squamata: Gekkonidae).

Thirty white-spotted geckos, Tarentola annularis, from the South Sinai desert in Egypt, were examined for helminth parasites. Spauligodon aspiculus was observed to infect 19 geckos with 63.33% as a prevalence of parasitic infection. The present nematode species is separated from congeners by morphological and metrical characteristics such as lateral alae, aspinose filamentous tail, and no spicule, and three pairs of caudal papillae with posterior pair excluded from envelopment by the caudal alae in the male worms, and knobbed eggs, and postbulbar vulva in females. It compared morphometrically with other Spauligodon species described previously and showed few differences in measurements. Molecular characterization based on the partial 28S rRNA nuclear ribosomal gene sequence showed that there was a close identity, up to 72%, with other sequences retrieved from GenBank. Phylogenetic analysis showed that the parasite sequence in conjunction with existing data facilitates the investigation of the placement of this pharyngodonid species within Oxyuridae. The present species is deeply embedded in the genus Spauligodon with close relationships to previously described Spauligodon nicolauensis (gb| JN619349.1, and JF829243.1) as more related sister taxa. This study highlights the importance of combining genetic and morphological data with taxonomy in pharyngodonid species.

Redescription of Gyrinicola japonica, a Tadpole-Endoparasitic Nematode from Japan, with Resurrection of the Family Gyrinicolidae (Nematoda: Oxyurina).

The taxonomic account of the tadpole-parasitic nematode Gyrinicola japonica Yamaguti, 1938, which is the type species of the genus, was reassessed based on syntypes and newly-collected specimens from the type locality. Our redescription of G. japonica addresses the erroneous original description of a spicule in this nematode, and emends the diagnosis of the species. Additionally, molecular phylogenetic trees based on nuclear 18S and 28S rDNA sequences revealed that G. japonica forms a distinctive lineage within the suborder Oxyurina, and this tadpole-specialist is phylogenetically close to the lizard-parasitic nematodes that belong to the family Pharyngodonidae. The results of morphological examination with the aid of molecular phylogenetic trees highlight the systematic uniqueness of this tadpole-parasitic group within Oxyurina, and Gyrinicolidae is accordingly resurrected as a distinctive oxyurinan family, with redefinition of the family and the genus Gyrinicola.

Unveiling patterns of genetic variation in parasite-host associations: an example with pinworms and Neotropical primates.

Patterns of genetic variation among populations can reveal the evolutionary history of species. Pinworm parasites are highly host specific and form strong co-evolutionary associations with their primate hosts. Here, we describe the genetic variation observed in four Trypanoxyuris species infecting different howler and spider monkey subspecies in Central America to determine if historical dispersal processes and speciation in the host could explain the genetic patterns observed in the parasites. Mitochondrial (cox1) and ribosomal (28S) DNA were analysed to assess genetic divergence and phylogenetic history of these parasites. Sequences of the 28S gene were identical within pinworms species regardless of host subspecies. However, phylogenetic analyses, haplotype relationships and genetic divergence with cox1 showed differentiation between pinworm populations according to host subspecies in three of the four Trypanoxyuris species analysed. Haplotype separation between host subspecies was not observed in Trypanoxyuris minutus, nor in Trypanoxyuris atelis from Ateles geoffoyi vellerosus and Ateles geoffoyi yucatanensis. Levels of genetic diversity and divergence in these parasites relate with such estimates reported for their hosts. This study shows how genetic patterns uncovered in parasitic organisms can reflect the host phylogenetic and biogeographic histories.

Integrative approach on Pharyngodonidae (Nematoda: Oxyuroidea) parasitic in reptiles: Relationship among its genera, importance of their diagnostic features, and new data on Parapharyngodon bainae.

The first integrative approach using sequences of two genes (18S and 28S rRNA) plus morphological and life history traits, was explored in Pharyngodonidae nematodes parasitic in reptiles. Additionally, first genetic characterization of Parapharyngodon bainae and new data on its morphology are given. This approach evaluated the phylogenetic relationships among genera within Pharyngodonidae, as well as the importance of their diagnostic morphological features. Specimens of P. bainae were collected from faecal pellets of the lizard Tropidurus torquatus in the State of Minas Gerais, Brazil. Nematodes were fixed for scanning electron microscopy and molecular procedures. Morphological observations revealed the accurate structures of cephalic end, of cloacal region in males, of vulva and eggs. Phylogenetic reconstructions were based upon four datasets: aligned sequences of the 18S, of the 28S, of both concatenated genes and of combined morphological and molecular datasets. Bayesian inference and maximum likelihood were performed to infer the phylogenies of molecular datasets and maximum parsimony to infer that of all-combined data. Pharyngodonid parasites of reptiles seem to configure two general monophyletic lineages, as previously assertions. Results also showed the monophyly of Spauligodon, Skrjabinodon and Parapharyngodon, as well as the clear separation between the latter and Thelandros. Combination of datasets improved nodal supports. Analysis of the all-combined datasets revealed the importance of vulval position and egg morphology as phylogenetic informative traits. However, characters of male caudal morphology appear as are highly homoplastic, and seem to be product of convergent evolution or multiple losses of ancestral traits. The closely-related Thelandros and Parapharyngodon are kept valid and their diagnosis should be based upon the position of the operculum in eggs (terminal or subterminal, respectively). Some inconsistencies in the scarce molecular and morphological databases were noted. Thus, new genetic data is required for further conclusions and current database must be evaluated with attention.

Molecular characterization of cosmopolitan and potentially co-invasive helminths of commensal, murid rodents in Gauteng Province, South Africa.

Concurrent studies of helminth parasites of introduced and native rodent species are few and miss the opportunity to identify potential co-invasive parasite species. This study employed molecular tools to infer the phylogeny and elucidate the origin of potentially co-invasive parasites of commensal, murid rodents by assessing introduced Rattus norvegicus, Rattus rattus, Rattus tanezumi, and native Mastomys coucha in Gauteng Province, South Africa. Genotypes of Nippostrongylus brasiliensis recovered from R. norvegicus are nearly identical to those recovered from elsewhere in the world. The pinworms, Aspiculurus tetraptera, recovered from introduced R. tanezumi and R. rattus, Syphacia muris recovered from R. tanezumi, and Syphacia obvelata recovered from indigenous M. coucha have affiliations to those recovered of laboratory rodents from the USA and China. Syphacia obvelata was previously only known as a commensal endoparasite of laboratory rodents, and the S. muris genotype recovered from R. tanezumi in this study shows an affiliation to a genotype recovered from the same host species in Indonesia which is part of the native range. The study emphasizes the need for surveillance of potential co-invasive species and contributes in documenting genetic diversity of endoparasites of well-known hosts.

Parasitic nematodes of the genus Syphacia Seurat, 1916 infecting Muridae in the British Isles, and the peculiar case of Syphacia frederici.

Syphacia stroma (von Linstow, 1884) Morgan, 1932 and Syphacia frederici Roman, 1945 are oxyurid nematodes that parasitize two murid rodents, Apodemus sylvaticus and Apodemus flavicollis, on the European mainland. Only S. stroma has been recorded previously in Apodemus spp. from the British Isles. Despite the paucity of earlier reports, we identified S. frederici in four disparate British sites, two in Nottinghamshire, one each in Berkshire and Anglesey, Wales. Identification was based on their site in the host (caecum and not small intestine), on key morphological criteria that differentiate this species from S. stroma (in particular the tail of female worms) and by sequencing two genetic loci (cytochrome C oxidase 1 gene and a section of ribosomal DNA). Sequences derived from both genetic loci of putative British S. frederici isolates formed a tight clade with sequences from continental worms known to be S. frederici, clearly distinguishing these isolates from S. stroma which formed a tight clade of its own, distinct from clades representative of Syphacia obvelata from Mus and S. muris from Rattus. The data in this paper therefore constitute the first record of S. frederici from British wood mice, and confirm the status of this species as distinct from both S. obvelata and S. stroma.

Getting there and around: Host range oscillations during colonization of the Canary Islands by the parasitic nematode Spauligodon.

Episodes of expansion and isolation in geographic range over space and time, during which parasites have the opportunity to expand their host range, are linked to the development of host-parasite mosaic assemblages and parasite diversification. In this study, we investigated whether island colonization events lead to host range oscillations in a taxon of host-specific parasitic nematodes of the genus Spauligodon in the Canary Islands. We further investigated whether range oscillations also resulted in shifts in host breadth (i.e., specialization), as expected for parasites on islands. Parasite phylogeny and divergence time estimates were inferred from molecular data with Bayesian methods. Host divergence times were set as calibration priors after a priori evaluation with a global-fit method of which individual host-parasite associations likely represent cospeciation links. Parasite colonization history was reconstructed, followed by an estimation of oscillation events and specificity level. The results indicate the presence of four Spauligodon clades in the Canary Islands, which originated from at least three different colonization events. We found evidence of host range oscillations to truly novel hosts, which in one case led to higher diversification. Contemporary host-parasite associations show strong host specificity, suggesting that changes in host breadth were limited to the shift period. Lineages with more frequent and wider taxonomic host range oscillations prior to the initial colonization event showed wider range oscillations during colonization and diversification within the archipelago. Our results suggest that a lineage's evolutionary past may be the best indicator of a parasite's potential for future range expansions.

PCR Testing of IVC Filter Tops as a Method for Detecting Murine Pinworms and Fur Mites.

We evaluated PCR testing of filter tops from cages maintained on an IVC system through which exhaust air is filtered at the cage level as a method for detecting parasite-infected and -infested cages. Cages containing 4 naïve Swiss Webster mice received 360 mL of uncontaminated aspen chip or α-cellulose bedding (n = 18 cages each) and 60 mL of the same type of bedding weekly from each of the following 4 groups of cages housing mice infected or infested with Syphacia obvelata (SO), Aspiculuris tetraptera (AT), Myocoptes musculinus (MC), or Myobia musculi (MB) and Radfordia affinis (RA; 240 mL bedding total). Detection rates were compared at 30, 60, and 90 d after initiating bedding exposure, by using PCR analysis of filter tops (media extract and swabs) and testing of mouse samples (fur swab [direct] PCR testing, fecal flotation, anal tape test, direct examination of intestinal contents, and skin scrape). PCR testing of filter media extract detected 100% of all parasites at 30 d (both bedding types) except for AT (α-cellulose bedding, 67% detection rate); identified more cages with fur mites (MB and MC) than direct PCR when cellulose bedding was used; and was better at detecting parasites than all nonmolecular methods evaluated. PCR analysis of filter media extract was superior to swab and direct PCR for all parasites cumulatively for each bedding type. Direct PCR more effectively detected MC and all parasites combined for aspen chip compared with cellulose bedding. PCR analysis of filter media extract for IVC systems in which exhaust air is filtered at the cage level was shown to be a highly effective environmental testing method.

Lack of genetic structure in pinworm populations from New World primates in forest fragments.

Microevolutionary processes in parasites are driven by factors related to parasite biology, host abundance and dispersal, and environmental conditions. Here, we test the prediction that isolation of host populations results in reduced genetic diversity and high differentiation among parasite populations. We conducted a population genetic analysis of two pinworms, Trypanoxyuris minutus and Trypanoxyuris atelis, commonly found parasitizing howler and spider monkeys in tropical rainforests across south-eastern Mexico, whose populations are currently isolated due to anthropogenic habitat loss and fragmentation. Mitochondrial DNA was employed to assess parasite genetic patterns, as well as to analyse their demography and population history. Both pinworm species showed high haplotype diversity but, unexpectedly, lower nucleotide diversity than that reported for other parasites. No genetic differentiation or population structure was detected in either pinworm species despite habitat loss, fragmentation and host isolation. Several scenarios are discussed that could help to explain the genetic panmixia found in both pinworm species, including higher than expected primate inter-fragment dispersal movements, and passive dispersal facilitating gene flow between parasite populations. The results suggest that large population sizes of parasites could be helping them to cope with the isolation and fragmentation of populations, delaying the effects of genetic drift. The present study highlights the complexity of the drivers that intervene in the evolutionary processes of parasites. Detailed genetic studies are needed, both in host and parasite populations, to assess the effects that habitat perturbation and environmental changes could have on the evolutionary dynamics of pinworms and primates.

Cholinergic signaling plasticity maintains viscerosensory responses during Aspiculuris tetraptera infection in mice small intestine.

Intestinal parasites alter gastrointestinal (GI) functions like the cholinergic function. Aspiculuris tetraptera is a pinworm frequently observed in laboratory facilities, which infests the mice cecum and proximal colon. However, little is known about the impact of this infection on the GI sensitivity. Here, we investigated possible changes in spontaneous mesenteric nerve activity and on the mechanosensitivity function of worm-free regions of naturally infected mice with A. tetraptera. Infection increased the basal firing of mesenteric afferent nerves in jejunum. Our findings indicate that nicotinic but not muscarinic receptors, similarly affect spontaneous nerve firing in control and infected animals; these axons are mainly vagal. No difference between groups was observed on spontaneous activity after nicotinic receptor inhibition. However, and contrary to the control group, during infection, the muscarinic signaling was shown to be elevated during mechanosensory experiments. In conclusion, we showed for the first time that alterations induced by infection of the basal afferent activity were independent of the cholinergic function but changes in mechanosensitivity were mediated by muscarinic, but not nicotinic, receptors and specifically by high threshold nerve fibers (activated above 20mmHg), known to play a role in nociception. These plastic changes within the muscarinic signaling would function as a compensatory mechanism to maintain a full mechanosensory response and the excitability of nociceptors during infection. These changes indicate that pinworm colonic infection can target other tissues away from the colon.

The Missing Fellow: First Description of the Trypanoxyuris pigrae Male (Nematoda: Oxyuridae), a Parasite of the Black Howler Monkey (Alouatta pigra) in Mexico.

The first morphological description of the male of Trypanoxyuris pigrae Solórzano-García, Nadler, and Pérez-Ponce de León, 2016 , is presented in this study. Morphological data are supported by molecular data. Specimens of T. pigrae were recovered after the necropsy of a roadkill black howler monkey (Alouatta pigra) in southeastern Mexico. Males of T. pigrae are characterized by having 3 notched lips and a long esophagus with a posterior bulb; they also show a single crested lateral alae, a single spicule, and 4 caudal papillae. Morphological features coincide with those of the previously described T. pigrae females, and molecular profiles confirmed species identification. Males of T. pigrae are very similar to those of Trypanoxyuris minutus, another species of pinworm that also parasitizes the black howler monkey, A. pigra; however, the shape of the lips represents a very reliable diagnostic feature. Because of this, detailed en face observations are recommended to discriminate between these pinworm species.

The diversity and evolution of nematodes (Pharyngodonidae) infecting New Zealand lizards.

Host-parasite co-evolutionary studies can shed light on diversity and the processes that shape it. Molecular methods have proven to be an indispensable tool in this task, often uncovering unseen diversity. This study used two nuclear markers (18S rRNA and 28S rRNA) and one mitochondrial (cytochrome oxidase subunit I) marker to investigate the diversity of nematodes of the family Pharyngodonidae parasitizing New Zealand (NZ) lizards (lygosomine skinks and diplodactylid geckos) and to explore their co-evolutionary history. A Bayesian approach was used to infer phylogenetic relationships of the parasitic nematodes. Analyses revealed that nematodes parasitizing skinks, currently classified as Skrjabinodon, are more closely related to Spauligodon than to Skrjabinodon infecting NZ geckos. Genetic analyses also uncovered previously undetected diversity within NZ gecko nematodes and provided evidence for several provisionally cryptic species. We also examined the level of host-parasite phylogenetic congruence using a global-fit approach. Significant congruence was detected between gecko-Skrjabinodon phylogenies, but our results indicated that strict co-speciation is not the main co-evolutionary process shaping the associations between NZ skinks and geckos and their parasitic nematodes. However, further sampling is required to fully resolve co-phylogenetic patterns of diversification in this host-parasite system.