Digenea Carus, 1863 represent a highly diverse group of parasitic platyhelminths that infect all major vertebrate groups as definitive hosts. Morphology is the cornerstone of digenean systematics, but molecular markers have been instrumental in searching for a stable classification system of the subclass and in establishing more accurate species limits. The first comprehensive molecular phylogenetic tree of Digenea published in 2003 used two nuclear rRNA genes (ssrDNA = 18S rDNA and lsrDNA = 28S rDNA) and was based on 163 taxa representing 77 nominal families, resulting in a widely accepted phylogenetic classification. The genetic library for the 28S rRNA gene has increased steadily over the last 15 years because this marker possesses a strong phylogenetic signal to resolve sister-group relationships among species and to infer phylogenetic relationships at higher levels of the taxonomic hierarchy. Here, we have updated the database of 18S and 28S rRNA genes until December 2017, we have added newly generated 28S rDNA sequences and we have reassessed phylogenetic relationships to test the current higher-level classification of digeneans (at the subordinal and subfamilial levels). The new dataset consisted of 1077 digenean taxa allocated to 106 nominal families for 28S and 419 taxa in 98 families for 18S. Overall, the results were consistent with previous higher-level classification schemes, and most superfamilies and suborders were recovered as monophyletic assemblages. With the advancement of next-generation sequencing (NGS) technologies, new phylogenetic hypotheses from complete mitochondrial genomes have been proposed, although their power to resolve deep levels of trees remains controversial. Since data from NGS methods are replacing other widely used markers for phylogenetic analyses, it is timely to reassess the phylogenetic relationships of digeneans with conventional nuclear rRNA genes, and to use the new analysis to test the performance of genomic information gathered from NGS, e.g. mitogenomes, to infer higher-level relationships of this group of parasitic platyhelminths.

The aim of this study was to evaluate the pattern of local immune cell infiltration in human cystic echinococcosis (CE) by identifying the subtypes of immune cells using immunohistochemistry (IHC). Fifty surgically removed hydatid cyst samples and surrounding tissues were collected from patients referred to Al-Zahra Hospital, Isfahan, Iran. IHC was performed on the surrounding host tissue of hydatid cysts using anti-human CD3, CD19, CD8, CD4, CD68, CD56, Ki-67 and Foxp3 (forkhead box P3) antibodies. The results were then compared to hepatocellular carcinoma and chronic hepatitis. In the host-tissue reaction site of liver hydatid cysts, a distinct pattern of local immune cell response, which outwardly consisted of a pack of the fibrous elements, a layer of palisading macrophages, an eosinophil-containing layer and a layer of accumulated lymphocytes, was observed. However, in some cases there were no positive cells for CD56+ natural killer cells and Foxp3+ regulatory T cells. The CD3+ T cells were the predominant inflammatory cells in all groups, followed by CD19+ B cells. It can be concluded that different immune cells are involved in the local response to human hydatid cysts.

Despite the seriousness of schistosomiasis, its treatment depends only on praziquantel (PZQ), which has begun to lose its efficacy against the emergent Schistosoma mansoni-resistant strains. Therefore, the discovery of a novel schistosomicidal drug is an urgent priority. This study was designed to evaluate treatment with Cucurbita pepo L. (pumpkin) seed oil (PSO) alone and combined with PZQ against S. mansoni in experimentally infected mice. The study involved five groups: GI was the normal control; GII was the infected control; GIII was treated with an oral dose of PZQ of 500 mg/kg/day for two successive days, starting in the sixth week post infection; GIV was treated with an oral dose of PSO of 50 mg/kg/day for four weeks, starting in the fourth week post infection; and GV was treated with combined PSO–PZQ. Worm burden, tissue egg load and oogram pattern were estimated, and the ultrastructure alterations were examined. Histopathological examination of granuloma diameters, collagen deposition (Picro Sirius red stain), and angiogenesis (immunohistochemical expression of CD34+) was conducted and serum liver enzymes were measured to assess the liver condition. Moreover, the oxidative stress was evaluated by determining the amounts of malondialdehyde and superoxide dismutase in liver homogenates. The results revealed significant changes in all the assessed parameters with PSO administration. However, PZQ was significantly more effective as an antiparasitic agent, whereas PSO was better in terms of fibrosis and oxidative stress. The most significant results were obtained in group V, which may be attributed to a synergy between PZQ and PSO, with antiparasitic, antioxidant and antifibrotic properties.

Knowledge of endemic helminths in a resource-limited country such as Nigeria is essential for their diagnosis, treatment and cost-effective control. In the present study, the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guideline was employed to determine the prevalence and geographical distribution of zoonotic helminths in food animals slaughtered in Nigerian abattoirs between 1970 and 2016. Pooled prevalence estimate (PPE) was determined by the random-effects model while heterogeneity was evaluated using the Cochran's Q-test. Results from 42 eligible studies reported across 19 Nigerian states revealed 85,466 cases of zoonotic helminths from 3,771,832 slaughtered food animals. Overall PPE was 2.27% (95% confidence interval (CI): 2.25, 2.28). PPEs for sub-groups ranged between 0.51% (95% CI: 0.46, 0.56) and 18.05% (95% CI: 17.12, 19.01) across regions, hosts, study periods and diagnostic methods. Ascaris suum had the highest pooled prevalence of 25.46% (95% CI: 24.04, 26.92). Overall prevalence estimates for cestodes, nematodes and trematodes were 0.60% (95% CI: 0.59, 0.61), 21.51% (95% CI: 20.73, 22.30) and 1.86% (95% CI: 1.84, 1.87), respectively. A high degree of heterogeneity 99.97% (95% CI: 2.25, 2.28, P: 0.000) was observed. Zoonotic helminths were prevalent in slaughtered food animals, with higher prevalence estimates in the north–central region, pigs and during the last decade reviewed. Ascaris suum was the most prevalent helminth, while Fasciola gigantica had the widest geographical distribution. It is envisaged that the present information will help in the formulation of disease-control policies, encourage on-farm good agricultural practices, and adequate hygiene and sanitation in abattoirs and meat-processing plants, with the aim of protecting public health.

Human toxocariasis, a worldwide parasitic disease, is caused by the larval stage of intestinal nematodes of dogs and cats, namely Toxocara canis and Toxocara cati. Human infection occurs by the accidental ingestion of embryonated eggs present in the soil, vegetables or on other contaminated surfaces, as well as via consumption of uncooked paratenic hosts, such as bird meat and giblets. The objective of this study was to evaluate the contamination of soil in public parks and playgrounds in Shiraz using microscopy and molecular methods. A total of 150 soil samples were collected from public parks and playgrounds in various areas of Shiraz, southern Iran. The samples were treated with saturated zinc sulphate solution, and Toxocara spp. eggs were detected by microscopic observation followed by nested polymerase chain reaction (PCR). To differentiate T. canis and T. cati eggs from each other, PCR restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS)-rDNA region by SalI endonuclease enzyme was used. PCR-sequencing was performed to confirm the results of the PCR-RFLP method. Based on the flotation results of the 150 soil samples, six (4%) were found to be positive for Toxocara spp. eggs, whereas nested-PCR showed 24 samples to be positive (16%). Based on the PCR-RFLP method and the sequence of the ITS-rDNA region, a total of 23 out of 24 isolates were confirmed as T. cati and one out of 24 as T. canis. The results showed a higher number of soil samples to be positive for Toxocara by the molecular method than microscopy, and higher T. cati infection in soil samples, which could have an important role in human infection with toxocariasis in this region.

The presence and distribution of various species of canine hookworms in Africa are poorly known. The main objective of this study, therefore, was to identify the hookworm species present in canine faecal samples from Morogoro, Tanzania, using molecular techniques. Faecal samples from 160 local dogs were collected and hookworm positive samples processed to recover larvae for further molecular characterization. DNA was extracted from pools of larvae from individual samples (n = 66), which were analysed subsequently using two different molecular approaches, polymerase chain reaction-linked restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR coupled with Sanger sequencing. The PCR-RFLP technique detected only the presence of the ubiquitous Ancylostoma caninum in the 66 samples. However, by species-specific PCR coupled with Sanger sequencing we identified ten samples with A. braziliense, two with Uncinaria stenocephala and five with A. ceylanicum. Thus, all four known species of canine hookworms were identified in Morogoro, Tanzania. To our knowledge this is the first report of the detection of the presence of U. stenocephala and A. ceylanicum in Africa using molecular techniques. In addition to their veterinary importance, canine hookworms have zoonotic potential and are of public health concern.

The genetic mechanisms of how free-living nematodes evolved into parasites are unknown. Current genetic model nematodes (e.g. Caenorhabditis elegans) are not well suited to provide the answer, and mammalian parasites are expensive and logistically difficult to maintain. Here we propose the terrestrial gastropod parasite Phasmarhabditis hermaphrodita as a new alternative to study the evolution of parasitism, and outline the methodology of how to keep P. hermaphrodita in the lab for genetic experiments. We show that P. hermaphrodita (and several other Phasmarhabditis species) are easy to isolate and identify from slugs and snails from around the UK. We outline how to make isogenic lines using ‘semi-natural’ conditions to reduce in-lab evolution, and how to optimize growth using nematode growth media (NGM) agar and naturally isolated bacteria. We show that P. hermaphrodita is amenable to forward genetics and that unc and sma mutants can be generated using formaldehyde mutagenesis. We also detail the procedures needed to carry out genetic crosses. Furthermore, we show natural variation within our Phasmarhabditis collection, with isolates displaying differences in survival when exposed to high temperatures and pH, which facilitates micro and macro evolutionary studies. In summary, we believe that this genetically amenable parasite that shares many attributes with C. elegans as well as being in Clade 5, which contains many animal, plant and arthropod parasites, could be an excellent model to understand the genetic basis of parasitism in the Nematoda.

The helminth endoparasites of many European amphibian species are often known exclusively from morphological descriptions. A molecular library of DNA sequence data linked to morphological identifications is still in its infancy. In this paper, we aim to contribute to such a library on the smooth newt Lissotriton vulgaris, the intermediate and definitive host of 31 helminth parasites, according to evidence published so far. Newts (n = 69) were collected at two study sites in western Germany and examined for the presence of helminths. A total of five helminth species were detected in 56 (81%) of the newts, but only one or two species infected a single host. Four out of five helminth species were identified morphologically and based on DNA sequences as Parastrigea robusta (metacercariae), Oswaldocruzia filiformis, Megalobatrachonema terdentatum (adults and larvae) and Cosmocerca longicauda, and the corresponding sequences were provided subsequently. Oswaldocruzia molgeta was confirmed to be a junior synonym of O. filiformis. Molecular data on a fifth species (a cosmocercid nematode) that could not be identified at species level were added to GenBank. These findings increased the molecular library on morphologically identified smooth newt parasites significantly, from 12 to 15 entries.

Larval trematodes are the main parasites of snails, and they play a crucial role because they usually castrate their snail hosts and can thus alter their population and community dynamics. This study involved a survey of seven gastropod species (Crepipatella dilatata, Fissurella radiosa, Nacella magellanica, Pareuthria fuscata, Siphonaria lessonii, S. lateralis and Trophon geversianus) parasitized by 12 trematode species (one hemiurid, one gymnophallid, two lepocreadiids, two microphallids, one notocotylid, two renicolids, one philophtalmid, one schistosomatid and one zoogonid) from southern Patagonia (47°S, 65°W), Argentina. Only F. radiosa was free of parasites. The study included the description of five new larvae, based on morphological and molecular information, and a comparison of the parasite diversity with that of a northern locality (42°S, 64°W), characterized by a lower mollusc diversity. Species richness and diversity of parasites were higher in the southern site. This suggests a correlation between the level of parasitism and the diversity of molluscs (first intermediate hosts), which is higher at the high-latitude site and seems to attract shorebirds, which disperse the digenean eggs and facilitate the completion of their life cycles. These results support the notion that parasitism is influenced by large-scale factors such as biogeographical patterns, and small-scale factors such as diversity or abundance of intermediate and definitive hosts.

A new species of Nippostrongylinae (Nematoda: Heligmonellidae), Stilestrongylus rolandoi n. sp., is described from specimens collected from the small intestine of the rodent Euryoryzomys russatus in the Atlantic Forest (Santo Amaro da Imperatriz, Santa Catarina state, southern Brazil). The genus Stilestrongylus includes 23 species, which parasitize rodents occurring in the Neotropical region. Stilestrongylus aureus (Durette-Desset & Sutton, 1985) from Argentina, S. azarai (Durette-Desset & Sutton, 1985) from Argentina, S. flavescens (Sutton & Durette-Desset, 1991) from Uruguay, S. franciscanus (Digiani & Durette-Desset, 2002) from Argentina, S. gracielae (Digiani & Durette-Desset, 2006) from Argentina, and S. oryzomysi (Sutton & Durette-Desset, 1991) from Argentina are closely related to Stilestrongylus rolandoi n. sp., all having caudal bursa patterns of types 1–4 in one of the lobes. Stilestrongylus rolandoi n. sp. is distinguished from the aforementioned species by its ray 6 being short in relation to rays 4 and 5, which are long and robust, and by having caudal bursa patterns of types 1–4 in both lobes. The new species has 27 ridges in the mid-body in males, and 24 in females, and has one of the highest ratios of spicule length to body length (21−33%) in this genus.

Four strains of entomopathogenic nematodes were isolated with a live trap method in southern Poland. The isolates were identified as Steinernema silvaticum based on morphological, morphometric and molecular data. Infective juveniles of Polish S. silvaticum isolates differ in body length from S. weiseri (951 vs 740 μm, respectively), and in the hyaline tail portion from S. kraussei (48 vs 38%, respectively). First-generation males of S. silvaticum are longer than those of S. kraussei, S. weiseri and S. ichnusae (1829 vs 1400, 1180 and 1341 μm, respectively). Males of S. silvaticum and a sister species S. kraussei can be distinguished by the distance from the anterior end to the nerve ring (142 vs 105 μm), spicule (66 vs 49 μm) and gubernaculum length (45 vs 33 μm), and the presence of a mucron. The analysis of internal transcribed spacer (ITS), D2-D3 and cox1 sequences of the tested nematodes revealed differences of 3–5%, 3% and 12–13%, respectively, from S. kraussei strains. The phylogeny of both nuclear and mitochondrial genes indicated close relationships of the Polish S. silvaticum isolates with S. kraussei, S. oregonense and S. cholashanense. The reproductive isolation of the studied isolates was confirmed by hybridization tests with other European feltiae–kraussei group representatives. This study has supplemented the original description of S. silvaticum with morphological and morphometric characterization of the first-generation males and females. This is also the first molecular study of this species based on a multi-gene approach.

An abattoir study was carried out between May and October 2016 to determine the parasitic causes of organ condemnation during meat inspection and to evaluate the attendant financial losses in Fako abattoirs, in the South-West region of Cameroon. Organs (liver, lungs, heart, tongue, kidney, spleen and intestine) were examined at meat inspection following standard procedures and the financial loss was estimated by considering the total weight of condemned organs and the price per kilogram of marketable organs, obtained from the local market. The organs of 1472 cattle were examined, of which 357 (24.38%) were condemned. The organs condemned because of parasitic infestations were the liver (333) and small intestine (24), and the infections were caused by flukes of Fasciola sp. and proglottids of Moniezia sp., respectively. Hydatid cysts and cysticerci were absent. The prevalence of fasciolosis and monieziosis was 22.62% and 1.63%, respectively. Condemnation deprived the region of 665.457 kg of meat, with an associated financial loss of CFA 1,330,902 (USD 2505), during the study period. Parasitic diseases worsen the food insecurity situation as they result in the withdrawal of a considerable amount of meat from the food chain. Fasciolosis, the leading parasitic cause of meat condemnation in Fako, is also zoonotic. It is therefore important that effective control measures are implemented countrywide against this parasitosis.

Although there have been occasional reports of rare and low-level trichinellae infestation in beavers, no human cases of beaver-associated trichinellosis have been described. This report presents a possible case of human trichinellosis linked to beaver meat. Increasing consumption of beaver meat necessitates raising awareness of this potential source of trichinellosis.

Dipetalonema caudispina (Molin, 1858) and D. gracile (Rudolphi, 1809) (Filarioidea: Onchocercidae) are two of six known species of filarial nematodes that parasitize Neotropical non-human primates. Adult filariae were collected from the thoracic and abdominal cavities of 38 of 44 specimens of Sapajus macrocephalus (Spix, 1823) and nine of ten specimens of Cebus albifrons (Humboldt, 1812) (Primates: Cebidae), distributed in the Yavarí-Mirín river basin and used locally for human consumption. Co-occurrence of D. caudispina and D. gracile is reported for the first time, with a prevalence of 18.5% (10 of 54 hosts examined). Our finding of D. caudispina and D. gracile in cebids from the Peruvian Amazon constitutes a new geographical record for both filariae, two new host records for D. caudispina, and the first report of D. gracile in S. macrocephalus. In addition, we provide morphometric data for D. caudispina, complementing the original description, as well as scanning electron microscopy details on the structure of the area rugosa and number of caudal papillae in males.

Susceptibility and tolerance to parasite infection vary greatly between host species. The congeneric isopod species Austridotea annectens and Austridotea lacustris often occur in sympatry and can both serve as intermediate hosts for the trematode Maritrema poulini. However, the intensity and prevalence of infections vary greatly in natural populations, with A. annectens often being heavily infected and A. lacustris rarely being infected. To shed light on the factors that may be involved in infection avoidance in A. lacustris, 100 isopods were collected and experimentally exposed to M. poulini cercariae. To examine for potential behavioural mechanisms, we used carbonated water as a paralysing agent to temporarily stop any movement by the isopods, and exposed paralysed isopods to cercariae. In the experiments, none of the individuals that were exposed to the parasite were found to be infected, although some cercariae seemed to have penetrated isopod hosts. Behavioural defence mechanisms do not seem to explain the different infection levels between A. lacustris and A. annectens, suggesting a physiological, possibly immunological, factor may be present. At the ecosystem level, this suggests a potential dilution effect caused by this low-competency host, and its effects on parasite transmission dynamics should be investigated.

Parascarosis is caused mainly by parasitic infections with Parascaris equorum and Parascaris univalens, the most common ascarid nematodes, in the small intestine of equines. Parascarosis often causes severe illness and even death in foals and yearlings. In this study, we obtained the complete sequence of the P. equorum mitochondrial (mt) genome and compared its organization and structure with that of P. equorum Japan isolate (nearly complete), and the complete mtDNA sequences of P. univalens Switzerland and USA isolates. The complete mtDNA genome of P. equorum China isolate is 13,899 base pairs (bp), making it the smallest of the four genomes. All four Parascaris mt genomes are circular, and all genes are transcribed in the same direction. The P. equorum mtDNA genome consists of 12 protein-coding genes, two ribosomal RNA genes, 22 transfer (t) RNA genes and one non-coding region, which is consistent with P. equorum Japan isolate and P. univalens Switzerland isolate but distinct from P. univalens USA isolate, which has 20 tRNA genes. Differences in nucleotide sequences of the four entire mt genomes range from 0.1–0.9%, and differences in total amino acid sequences of protein-coding genes are 0.2–2.1%. Phylogenetic analyses showed that the four Parascaris species clustered in a clade, indicating that P. equorum and P. univalens are very closely related. These mt genome datasets provide genetic evidence that P. equorum and P. univalens may represent the same species, which will be of use in further studies of the taxonomy, systematics and population genetics of ascarids and other nematodes.