A new entomopathogenic nematode species of Heterorhabditis, described as H. pakistanense n. sp., was isolated from soil samples around the roots of grass at Malir, Karachi, Sindh, Pakistan. The new species is characterized morphologically by features of males: body size 819 μm (720–1013 μm), D% ((distance from anterior end to excretory pore divided by pharynx length) × 100) 119 (110–126), SW% ((spicule length divided by anal body diameter) × 100) 156 (144–191), GS% ((gubernaculum length divided by spicule length) × 100) 58 (48–65) and variations in the number of bursal papillae of the terminal group: 8th and 9th papillae sometimes absent on both sides, sometimes eight papillae present on the right side whereas six papillae present on the left side. On the right side the arrangement of papillae is 1 + 2 + 3 + 2 whereas on the left side it is 1 + 2 + 3. The hermaphrodite has a prominent post-anal swelling and a conoid tail 82 μm (64–95 μm) long with a pointed terminus. Hermaphrodites of H. pakistanense n. sp. can be distinguished from all species of Heterorhabditis except H. downesi by having a mucronate tail. Infective juveniles have a medium-sized body (581 μm (558–624 μm)), long pharynx (117 μm (113–125 μm)), ensheathed tail (99 μm (95–110 μm)) and E% ((distance from anterior end to excretory pore divided by tail length) × 100) 100 (95–107). The new species can be distinguished from all species of Heterorhabditis by the absence of the 7th, 8th and 9th bursal papillae. Heterorhabditis pakistanense is further characterized by the internal transcribed spacer (ITS) and the D2D3 region of the 28S rDNA gene. The closest species H. indica, H. gerrardi, H. amazonensis and H. noenieputensis being separated by 9, 7, 66 and 15 bp, respectively, in the ITS region. Molecular phylogenetic trees based on sequences of ITS rDNA, D2D3 regions and the mitochondrial 12S rRNA gene support the description of H. pakistanense as a new species.

Studies of amphibian macroparasites are relevant for the investigation of parasite community ecology and disease dynamics. Here, the parasite communities of five anuran species (Hypsiboas raniceps, Phyllomeduza azurea, Pseudis paradoxa, Leptodactylus fuscus and Leptodactylus podicipinus) are described from two habitats with different levels of preservation (pasture versus nature reserve). Specifically, we used mixed-effect models to test whether helminth species richness, prevalence and abundance differ between the two host collection sites. A total of 120 anuran individuals and 25 helminth parasite taxa were collected. Helminth communities differed between collection sites and among host species. In general, helminth species richness, prevalence and abundance were higher in hosts collected in the pond from the nature reserve. In all, these data help fill the gap in parasite biodiversity knowledge in a changing area, within a sensitive group of vertebrate hosts.

Resolving complex life cycles of parasites is a major goal of parasitological research. The aim of this study was to analyse the life cycle of two species of the genus Profilicollis, the taxonomy of which is still unstable and life cycles unclear. We extracted individuals of Profilicollis from two species of crustaceans (intermediate hosts) and four species of seagulls (definitive hosts) from sandy-shore and estuarine habitats along the south-east Pacific coast of Chile. Mitochondrial DNA analyses showed that two species of Profilicollis infected intermediate hosts from segregated habitats: while P. altmani larvae infected exclusively molecrabs of the genus Emerita from fully marine habitats, P. antarcticus larvae infected the crab Hemigrapsus crenulatus from estuarine habitats. Moreover, P. altmani completed its life cycle in four seagulls, Chroicocephalus maculipennis, Leucopheus pipixcan, Larus modestus and L. dominicanus, while P. antarcticus, on the other hand, completed its life cycle in the kelp gull L. dominicanus. Accordingly, our results show that two congeneric parasites use different and spatially segregated species as intermediate hosts, and both are capable of infecting one species of definitive hosts. As such, our analyses allow us to shed light on a complex interaction network.

Explanatum explanatum flukes, liver amphistomes of ruminants, cause significant economic loss in the livestock industry by inducing severe liver damage. A total of 66 flukes from 26 buffaloes and 7 cattle in four different geographic areas of Bangladesh and 20 flukes from 10 buffaloes in the Chitwan district of Nepal were subjected for analysis. The sequences (442 bp) of the second internal transcribed spacer (ITS2) of ribosomal DNA and the variable fragments (657 bp) of mitochondrial nicotinamide dehydrogenase subunit 1 (nad1) of E. explanatum flukes from Bangladesh and Nepal were analysed. The aim of this study was molecular characterization of the flukes and to elucidate their origin and biogeography. In the ITS2 region, two genotypes were detected among the flukes from Bangladesh, while flukes from Nepal were of only one genotype. Phylogenetic analyses inferred from the nad1 gene revealed that at least four divergent populations (groups I–IV) are distributed in Bangladesh, whereas two divergent populations were found to be distributed in Nepal. Fst values (pairwise fixation index) suggest that Bangladeshi and Nepalese populations of group I to IV are significantly different from each other; but within groups III and IV, the populations from Bangladesh and Nepal were genetically close. This divergence in the nad1 gene indicates that each lineage of E. explanatum from diverse geography was co-adapted during the multiple domestication events of ruminants. This study, for the first time, provides molecular characterization of E. explanatum in Bangladesh and Nepal, and may provide useful information for elucidating its origin and dispersal route in Asia.

Molecular and morphological characterization of two species of Cucullanidae from freshwater fish in Brazil are provided, one of which represented an undescribed taxon. Cucullanus opisthoporus n. sp. was collected in Cichla melaniae from River Xingu, State of Pará, and in C. pinima from River Jamarí, State of Rondônia. Nematodes referable to Cucullanus grandistomis were collected in Oxydoras niger from River Xingu. The new species has an appendage in the tail tip, ventrally covered by small spines, which is an exclusive feature of Cucullanus tucunarensis. However, C. tucunarensis differs from C. opisthoporus n. sp. based upon the relative position of deirids and the excretory pore, which are more posterior from the oesophageal end in the new species. Observations of C. tucunarensis type specimens also revealed features that were wrongly or not reported in the original description. Type specimens of C. grandistomis were observed, although they were poorly preserved. After evaluation of newly collected specimens of C. grandistomis, features unreported in the original description were observed for the first time, including the presence of an intestinal caecum. Thus, C. grandistomis was transferred to Dichelyne. Sequences of the 18S and 28S rRNA genes revealed high genetic similarity between the specimens of C. opisthoporus n. sp. from the two different hosts as well as their genetic distance from Dichelyne grandistomis n. comb. Phylogenetic reconstructions using representatives of Cucullanidae suggested the artificiality of the current morphological system adopted to separate the genera, since most genera were not monophyletic, although the availability of genetic data is still fragmented.

There is little information on the phylogenetic position and life cycle of family Kathlaniidae. Falcaustra araxiana is a member of this family which infects the large intestine of the European pond turtle (Emys orbicularis). In the present study, morphological data and molecular analyses based on the 18S rDNA were performed on different types of F. araxiana originating from the large intestine and gastric nodules in the turtle. Morphological data revealed both larvae and adult stages in the gastric nodules. In addition, all nematodes recovered in the large intestine were adult worms. GenBank accession numbers KM200715 and KM200716 were provided for adult F. araxiana located in the intestine and stomach, respectively, of E. orbicularis. The results of sequencing proved that these two types are completely similar. Accordingly, it can be hypothesized that nodule formation is a part of the life cycle of the parasite or a survival strategy. Furthermore, F. araxiana develops to the adult stage in the gastric mucosa prior to migrating to the large intestine. Phylogenetic analysis revealed that F. araxiana unexpectedly branched away from other members of the superfamily Seuratoidea (Truttaedacnitis truttae, Cucullanus robustus and C. baylisi) and showed a closer relationship with Paraquimperia africana, a member of the Ascaridoidea. It seems that phylogenetic reconstruction for the present parasite needs more detailed morphology, life cycle and molecular studies.

Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) is described from the lungs of the ‘Brazilian lancehead’ Bothrops moojeni (Hoge, 1966) from the savannah in São Paulo State, Brazil. The new species is the eighth species of Serpentirhabdias described in the Neotropical region, and differs from other species mainly by a combination of characters: lips slightly notable, presence of fine striations at posterior ends, presence of two parallel lines with intercalated pores, a pore-shaped phasmid situated at the level of the anal aperture and another two in the posterior half of the tail. It is the first species of Serpentirhabdias reported in this snake host and the second species of this genus found parasitizing South American viperidian snakes. Molecular phylogenetic analysis using ribosomal (ITS and 28S partial) genes confirms Serpentirhabdias viperidicus n. sp. as a new species that clustered in the Serpentirhabdias clade, sister taxon to Serpentirhabdias fuscovenosa and Serpentirhabdias elaphe. This is the first description of Serpentirhabdias species from Brazil using molecular approaches and morphological characters to confirm the monophyly of this recent genus.

The present investigation aimed to evaluate the extent to which maternal filarial infection influences IgG subclass immune responses in the cord blood of neonates. Prevalence of antigenaemia was detected using an Og4C3 assay. Filaria-specific IgG subclasses against excretory/secretory antigens were measured by ELISA. Transplacental transfer of circulating filarial antigen (CFA) was observed from 34.8% of CFA-positive mothers to their respective cord bloods. Filaria-specific IgG1, IgG2 and IgG4 responses were significantly higher among cord bloods of infected mothers compared to cord bloods of uninfected mothers. In contrast, the IgG3 response was significantly higher among cord bloods of uninfected mothers. The study shows that transplacental transfer of filarial antigens and filaria-specific IgG4 occurs more in mothers having high worm burdens, and transfer of filaria-specific IgG3 occurs more in the cord blood of uninfected mothers. The findings of the study provide evidence for the development of prenatal sensitization to filarial antigens in utero, and high filaria-specific IgG4 in cord blood may serve as a marker for in-utero sensitization.

Saccocoelioides olmecae n. sp. is described from specimens recovered from the intestine of the fat sleeper Dormitator maculatus (Bloch) (Perciformes: Eleotridae) collected in six localities along the coast of the Gulf of Mexico. The new species is mainly distinguished from the other three described species of Saccocoelioides Szidat, 1954 from North and Middle America (i.e. S. sogandaresi Lumsden, 1963, S. chauhani Lamothe-Argumedo, 1974 and S. lamothei Aguirre-Macedo & Violante-González, 2008) by having an elongated body, a sac-like caecum, a uterus that extends to the first third of body and by having vitelline follicles longitudinally elongated reaching the posterior end of the body. Sequences of the large subunit (LSU) of the ribosomal DNA, including the domain D1–D3, and the internal transcribed spacer 2 (ITS2) were used independently and concatenated to corroborate the morphological distinction among S. olmecae n. sp., S. chauhani and S. lamothei from freshwater and brackish-water fish from Middle America. The genetic divergence estimated among the three species of Saccocoelioides was very low: 1% for LSU and from 1 to 4% for ITS2. Maximum likelihood and Bayesian inference analyses for each dataset and both datasets combined revealed that S. olmecae n. sp. represents an independent clade with moderate bootstrap support and posterior probabilities. This is the third species of Saccocoelioides described in Mexico, and the 17th species from the Americas.

Morphological analysis of lungworms collected among Caprinae from Uzbekistan resulted in the identification of four species of Protostrongylidae: Protostrongylus rufescens, Protostrongylus hobmaieri, Spiculocaulus leuckarti and Cystocaulus ocreatus. The following species were recorded as definitive hosts: Ovis aries, Ovis ammon, Ovis vignei, Capra hircus, Capra falconeri and Capra sibirica. The prevalence of P. rufescens reached 45.3%, followed by S. leuckarti and C. ocreatus with 31.7% and P. hobmaieri with 16.9%. The sex ratio ranged between 1:3.1 and 1:6.2, with P. hobmaieri showing the strongest predominance of females over males. The prevalence of infection of small ruminants with protostrongylid nematodes increased with the age of the hosts. Protostrongyles use terrestrial gastropods as intermediate hosts, and infective larvae were found in the species Vallonia costata, Gibbulinopsis signata, Pupilla muscorum, Pseudonapaeus albiplicata, Pseudonapaeus sogdiana, Leucozonella ferghanica, Xeropicta candacharica, Candaharia levanderi and Macrochlamys sogdiana. Xeropicta candacharica was the most abundant gastropod and had the highest prevalence of infection with protostrongylids. Adult X. candacharica had a significantly higher infection intensity than juveniles. The epidemiology of protostrongylid infections is dynamic and subject to considerable changes. Further characterization of the interaction of protostrongylid parasites with their terrestrial gastropods as intermediate hosts and Caprinae as definitive hosts is required to understand these processes and to monitor the effects of changing ecological contexts.

Haplorchis taichui is an intestinal heterophyid fluke that is pathogenic to humans. It is widely distributed in Asia, with a particularly high prevalence in Indochina. Previous work revealed that the lack of gene flow between three distinct populations of Vietnamese H. taichui can be attributed to their geographic isolation with no interconnected river basins. To test the hypothesis that interconnected river basins allow gene flow between otherwise isolated populations of H. taichui, as previously demonstrated for another trematode, Opisthorchis viverrini, we compared the genetic structures of seven populations of H. taichui from various localities in the lower Mekong Basin, in Thailand and Laos, with those in Vietnam, using the mitochondrial cytochrome c oxidase subunit 1 (COX1) gene. To determine the gene flow between these H. taichui populations, we calculated their phylogenetic relationships, genetic distances and haplotype diversity. Each population showed very low nucleotide diversity at this locus. However, high levels of genetic differentiation between the populations indicated very little gene flow. A phylogenetic analysis divided the populations into four clusters that correlated with the country of origin. The negligible gene flow between the Thai and Laos populations, despite sharing the Mekong Basin, caused us to reject our hypothesis. Our data suggest that the distribution of H. taichui populations was incidentally associated with national borders.

The family Deletrocephalidae consists of three species, from two genera: Deletrocephalus dimidiatus, D. cesarpintoi and Paradeletrocephalus minor, which differ from one another in terms of the buccal capsule structure, the dorsal ray from the male copulatory bursa and the length of the vagina in the female. All these species are parasites of the gastrointestinal tract of the rhea, Rhea americana. Only D. dimidiatus was reported to be a parasite of the other rhea species, Darwin's rhea, R. pennata. Currently, there are no studies on the ultrastructure and biology of these parasites or their pathogenicity in these birds. Thus, the present study aimed to examine the ultrastructure and add more details of the morphology of D. dimidiatus from R. americana. Four adult rheas were necropsied, and the gastrointestinal tract was collected and examined for the presence of parasites. Nematodes were analysed by optical and scanning electron microscopy (SEM). All birds were parasitized by D. dimidiatus. Some morphological and morphometric characters were observed that differ from reports of other studies of this species by other authors. In addition, new details were added, well described by SEM, and a key to the species of the subfamily Deletrocephalinae is given.

Haemonchus contortus and Haemonchus placei are among the major parasites of small ruminants and cattle. Although infection with these nematodes is host-specific, with H. placei predominating in cattle and H. contortus in sheep, cross-infections are observed in areas where both parasites are sympatric, and hybrid offspring can occur. Therefore, a fast and precise method is required for differentiating the parasites. Identification based on spicule morphometry is the most common technique for differentiating Haemonchus species. However, because these measurements overlap between species, morphological analysis is insufficient for differentiating between helminth species. In this work, we present a reliable, conventional polymerase chain reaction (PCR)-based method that uses two species-specific primer pairs to differentiate between H. contortus and H. placei specimens and their hybrids. Each primer pair produces one single and distinct amplification band for each species, which enables the detection of hybrid specimens. These primer pairs were validated by testing eight different populations of H. contortus, H. placei and hybrids.

Oncicola venezuelensis Marteau, 1977 was found parasitizing adults of Leopardus pardalis (Linnaeus) found dead in Serra da Capivara National Park, Piauí state, Brazil, a new geographical locality reported for the species. The diversity of Oncicola Travassos, 1916 species is large, but genetic data are scarce. This article presents the results of genetic, morphological and ultrastructural studies carried out for taxonomic purposes. The first ultrastructural view showed a globular, short proboscis with 36 hooks, divided into six longitudinal rows of six hooks each. Hooks differ in size and shape: hooks I, II and III have a ‘chisel-shaped’ tip. The genetic profile included new sequences of ribosomal DNA ITS1, 5.8S and ITS2, and partial 28S rRNA regions. The results of maximum-likelihood tree analyses for each region showed Oligacanthorhynchidae Southwell et Macfie, 1925 close to Gigantorhynchidae Hamann, 1892 (supported >91%). Both use mammals and birds as definitive hosts. Morphological and ultrastructural studies combined with genetic analysis shed more light on the diversity of Oncicola species.

Thorny-headed acanthocephalan worms of the genus Profilicollis are widely distributed in the oceans of the world and present complex life cycles with intermediate and definitive hosts. The genus is still poorly known, with an unstable taxonomy and, for most species, incompletely characterized geographical distributions. In this study, based on molecular and morphological evidence, we report that the species Profilicollis altmani is also distributed along the South American Atlantic coast, using the mole crab Emerita brasiliensis as an intermediate host. As such, our record shows that P. altmani has a Pan-American distribution where five species of Emerita are utilized as intermediate hosts.

Three new species of Hysterothylacium Ward & Magath, 1917, including H. australe, H. kajikiae and H. brucei, from Australian marine fish are described and illustrated by light microscopy followed by genetic characterization of their first and second internal transcribed spacers (ITS-1 and ITS-2, respectively). This is the first study reporting ITS sequence data for adult Hysterothylacium spp. in Australia, which provides an insight into the identification of some of the Hysterothylacium larval types. Alignment of ITS sequences of these species with Hysterothylacium larval types previously reported in Australia showed that fourth-stage Hysterothylacium larval type XI from Seriola lalandi and third-stage Hysterothylacium larval type X from Sphyraene novae-hollandiae are identical with ITS sequences of H. australe, suggesting that these fish are natural intermediate/paratenic hosts of H. australe.

Although the efficacy of triclabendazole treatment against Fasciola gigantica has been shown before in cattle and buffaloes, there appears to be no published report on the efficacy of triclabendazole in impala or other antelope species. As part of a health monitoring programme at Mulungushi International Conference Centre, Lusaka, Zambia, a coprological examination was undertaken to investigate the helminthological status of captive impala (Aepyceros melampus). Of 39 fresh coprological samples, 46% contained F. gigantica. The source of infection was identified to be a fountain within the grounds of the conference centre. Lymnaea natalensis, collected from the study site, were induced to shed cercariae, and were thus confirmed as the snail intermediate host. In managing this disease, triclabendazole at 6 mg/kg was administered together with the feed bait. Water from the pond was drained; vegetation within it cleared and the pond allowed to dry for 1 week before water was replaced. Three weeks post-treatment, faecal examination revealed that the Fasciola had been cleared. To the best of our knowledge, this is a first case of Fasciola spp. infection reported in captive wild impala in Zambia and provides evidence that triclabendazole may be delivered to free-ranging antelope using medicated bait.