Tracheophilus cymbius (Trematoda: Cyclocoelidae) is a common tracheal fluke of waterfowl, causing serious loss in the poultry industry. However, taxonomic identification of T. cymbius remains controversial and confused. Mitochondrial (mt) genomes can provide genetic markers for the identification of closely related species. We determined the mt genome of T. cymbius and reconstructed phylogenies with other trematodes. The T. cymbius mt genome is 13,760 bp in size, and contains 12 protein-coding genes (cox 1–3, nad 1–6, nad 4L, cyt b and atp 6), 22 transfer RNA (tRNA) genes, two ribosomal RNA genes and one non-coding region. All are transcribed in the same direction. The A + T content is 62.82%. ATG and TAG are the most common initiation and termination codons, respectively. Phylogenetic analyses of concatenated nucleotide sequences show T. cymbius grouping in suborder Echinostomata, and clustering together, with high statistical support, as a sister taxon with Echinochasmus japonicus (Echinochasmidae), the two forming a distinct branch rooted to the ancestor of all Echinostomatidae and Fasciolidae species. This is the first report of the T. cymbius mt genome, and the first reported mt genome within the family Cyclocoelidae. These data will provide a significant resource of molecular markers for studying the taxonomy, population genetics and systematics of trematodes.

The production of tambaqui Colossoma macropomum has been undergoing financial losses due to parasitic infection by the acanthocephalan Neoechinorhynchus buttnerae, raising an alert for aquaculture in South America. The lack of adequate treatment and use of unlicensed chemicals encourages research for alternative solutions with minimal side effects. The objectives of this study were to evaluate the in vitro antiparasitic potential of commercial nutraceutical products (Natumix® and BioFish®) against N. buttnerae and to assess the respective in vivo toxic effects on the host tambaqui. For in vitro assays, parasitized fish were necropsied for acanthocephalans sampling. The parasites were exposed to three concentrations (0.078, 0.313 and 1.25 mg/ml) of each product, as well as controls (one without product and another with a solubilizer). For the in vivo acute toxicity test, juvenile fish (<0.1 g) were exposed to five increasing concentrations of each product. Mortality of tambaqui was recorded at 24, 48, 72 and 96 h. The estimated lethal concentration (LC) for 10, 50, 90 and 99% of fish was determined to classify the toxicity of the products on the target species. After in vitro efficacy tests, the highest concentrations (1.25 mg/ml) caused 100% mortality of the parasites in both products, but only Natumix® caused 100% mortality using the intermediate concentration (0.313 mg/ml) after 24 h. According to the acute toxicity result, the LC50 classified the nutraceutical products as slightly toxic for tambaqui. The tested products had a parasiticidal effect on N. buttnerae, and the toxicity test showed that both products have therapeutic potential when added to the diet.

Helminth and protozoan infections are responsible for important diseases in grazing sheep, which can be especially threatening in an autochthonous breed at risk of extinction like the Churra Galega Mirandesa Portuguese sheep breed. The aim of the present study was to determine the diversity, prevalence and burden of gastrointestinal parasites in these sheep and to assess the effects of deworming practices, cohabiting animals on the farm and feed management. Coprological qualitative and quantitative analysis (flotation, natural sedimentation and McMaster method) were used to identify and quantify gastrointestinal parasites and a questionnaire was designed and applied. A total of 512 faecal samples were collected from 49 flocks, and 49 replies to the questionnaire were received. Parasites were identified in 100% of the flocks, and in 97% of the samples. The genera or species that have been morphologically identified were: strongyle-type, Nematodirus spp., Skrjabinema spp., Moniezia expansa, Moniezia benedeni, Trichuris spp., Capillaria spp., Eimeria spp., Dicrocoelium spp. and Fasciola hepatica. This is the first report in Portugal of Skrjabinema spp. The burden of parasites’ oocysts and eggs per gram in faecal samples ranged, respectively, from 50 to 17,550 for Eimeria spp., and from 50 to 6250 for strongyle-type eggs. Factors affecting parasitic infections were evaluated using a multivariate logistic regression. Grazing time and a lack of anthelmintic treatment were positively associated with Nematodirus spp. infection. This study showed that there is a high prevalence and diversity of gastrointestinal parasites in the Churra Galega Mirandesa sheep breed.

The human–animal bond is beneficial for human health, but companion animals also pose a potential threat as vectors of zoonotic parasites, especially in urban areas where both human and dog densities are high. However, the knowledge about parasitic spillover in the urban environment is relatively scarce. The aim of the present study was to reveal which factors determine parasitic contamination in Estonian towns and provide up-to-date information about intestinal parasites of the Estonian dog population. In total, 657 samples of dog excrement was collected over one year of investigation from five towns in Estonia. Generalized linear mixed models were used to evaluate factors predicting infection risk in urban areas. In general, infection risk and intensity models predicted higher infection with endoparasites for small dogs in smaller towns, especially in apartment-house districts and in potential hazard zones. Helminth eggs and Giardia/Cystoisospora oocysts were detected in 64 samples, with an overall prevalence of 9.8%.

The superfamily Pronocephaloidea Looss, 1899 comprises digeneans occurring in the gut and respiratory organs of fishes, turtles, marine iguanas, birds and mammals. Although many life cycles are known for species of the Notocotylidae Lühe, 1909 maturing in birds and mammals, relatively few are known for the remaining pronocephaloid lineages. We report the cercariae of five pronocephaloid species from marine gastropods of the Queensland coast, Australia. From Lizard Island, northern Great Barrier Reef, we report three cercariae, two from Rhinoclavis vertagus (Cerithiidae) and one from Nassarius coronatus (Nassariidae). From Moreton Bay, southern Queensland, an additional two cercariae are reported from two genotypes of the gastropod worm shell Thylacodes sp. (Vermetidae). Phylogenetic analysis using 28S rRNA gene sequences shows all five species are nested within the Pronocephaloidea, but not matching or particularly close to any previously sequenced taxon. In combination, phylogenetic and ecological evidence suggests that most of these species will prove to be pronocephalids parasitic in marine turtles. The Vermetidae is a new host family for the Pronocephaloidea.

The association between eosinophil count and cholelithiasis among people with Clonorchis sinensis infection is still uncertain. We conducted a cross-sectional study to investigate the associations among Clonorchis sinensis infection, eosinophil count and cholelithiasis. The study included 4628 participants from January to December 2018. The levels of eosinophil count were divided into four groups according to the quartiles of eosinophil count. Spearman's rank correlation was performed to assess the association between eosinophil counts and Clonorchis sinensis egg counts. Multiple regression analysis was performed to evaluate the relationships among C. sinensis infection, eosinophil count and cholelithiasis after adjusting for three models. The prevalence of C. sinensis infection was 38.72% (1792/4628), and the prevalence of cholelithiasis was 6.03% (279/4628). The infection rate of C. sinensis was higher in the cholelithiasis group than in the non-cholelithiasis group (63.08% vs. 37.16%, P < 0.001). Significant differences were found among various eosinophil count quartiles for C. sinensis infection, body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyltranspeptidase (γ-GT), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), creatinine (CRE), blood urea nitrogen (BUN), uric acid (UA) and non-alcoholic fatty liver disease (NAFLD) (all P < 0.001). A significant positive correlation was found between eosinophil count and log-transformed C. sinensis egg count (r = 0.9477, P < 0.001). Multiple logistic regression analysis revealed that light and moderate intensities of C. sinensis infection were associated with cholelithiasis (P < 0.01 and P < 0.001, respectively), and C. sinensis infection with eosinophil count ranging from 0.05 to 0.5 × 109/l were associated with cholelithiasis (P < 0.05). In conclusion, our findings suggest that the light and moderate infections of C. sinensis with eosinophil count ranging from 0.05 to 0.5 × 109/l may be associated with a higher risk of cholelithiasis.

The adult and metacercaria life stages of a new species of the microphallid genus Atriophallophorus Deblock & Rosé, 1964 are described from specimens collected at Lake Alexandrina (South Island, New Zealand). In addition to molecular analyses of ribosomal and mitochondrial genes, metacercariae of Atriophallophorus winterbourni n. sp. from the snail host Potamopyrgus antipodarum (Gray) were grown in vitro to characterize internal and external morphology of adults using light and scanning electron microscopy and histological techniques. Atriophallophorus winterbourni n. sp. is readily distinguishable from Atriophallophorus coxiellae Smith, 1973 by having a different structure of the prostatic chamber, sub-circular and dorsal to genital atrium, rather than cylindrical, fibrous, elongate and placed between the seminal vesicle and the genital atrium. The new species is most similar to Atriophallophorus minutus (Price, 1934) with regards to the prostatic chamber and the morphometric data, but possesses elongate-oval testes and subtriangular ovary rather than oval and transversely oval in A. minutus. Phylogenetic analyses including sequence data for A. winterbourni n. sp. suggested a congeneric relationship of the new species to a hitherto undescribed metacercariae reported from Australia, both forming a strongly supported clade closely related to Microphallus and Levinseniella. In addition, we provide an amended diagnosis of Atriophallophorus to accommodate the new species and confirm the sinistral interruption of the outer rim of the ventral sucker caused by the protrusion of the dextral parietal atrial scale at the base of the phallus.

Echinococcus granulosus sensu stricto is regarded to have the highest zoonotic potential of all Echinococcus taxa. Globally, human infection due to this species constitutes over 88.44% of the total cystic echinococcosis (CE) burden. Here, we report a CE infection in a Nigerian camel caused by E. granulosus G1 genotype. To the best of our knowledge, this report is the first encounter of the G1 genotype in the West Africa sub-region where the G6 genotype is reportedly prevalent, suggesting that the epidemiology of this highly zoonotic group could have a wider host range and distribution in the sub-region, and emphasizes the need for further investigation into the genetic diversity of Echinococcus spp. in Nigeria and across the sub-region.

Human strongyloidiasis is a deleterious gastrointestinal disease mainly caused by Strongyloides stercoralis infection. We aimed to study the possible transmission of S. stercoralis between humans and pet animals. We isolated Strongyloides from humans and domestic dogs in the same rural community in north-east Thailand and compared the nucleotide sequences of derived worms using portions of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and 18S ribosomal RNA (18S rRNA) genes. Twenty-eight sequences from the 18S rRNA gene were obtained from worms derived from humans (n = 23) and dogs (n = 5), and were identical with S. stercoralis sequences (from Thailand, Cambodia, Lao PDR and Myanmar) published in the GenBank database. The 28 cox1 sequences from humans and dogs showed high similarity to each other. The available published cox1 sequences (n = 150), in combination with our 28 sequences, represented 68 haplotypes distributed among four clusters. The 28 samples from the present study represented eight haplotypes including four new haplotypes. Dogs and humans shared the same haplotypes, suggesting the possibility of zoonotic transmission from pet dogs to humans. This is of concern since dogs and humans live in close association with each other.

The efficacy of eight essential oils (EOs) (Solidago canadensis, Eucalyptus globulus, Pelargonium asperum, Ocimum basilicum, Thymus vulgaris, Mentha piperita, Cymbopogon citratus and Cymbopogon martinii) against gastrointestinal nematodes (GINs) was evaluated using eggs collected from naturally infected cattle and cultured infective larvae (L3). The larvae species cultured from the faecal samples and subjected to two in vitro tests were Haemonchus spp. (55.5%), Trichostrongylus spp. (28.0%), Cooperia spp. (15.0%) and Oesophagostomum spp. (1.5%). The genus of EO Cymbopogon (C. citratus and C. martinii) showed the highest anthelmintic activity at the dose of 8.75 mg/ml, for the egg hatch, the larval migration and mortality assays. All of the EOs tested reduced egg hatching to rates <19.0%, compared to the controls (water and water + Tween 20) that had rates >92.0%. Cymbopogon citratus and C. martinii treatments resulted in 11.6 and 8.1% egg hatch, had the lowest migration of larvae through sieves, 60.5 and 54.9%, and the highest mortality rates, 63.3 and 56.3%, respectively. Dose–response tests showed that EO from C. citratus had the lowest larval LC50 and migration inhibition concentration (IC50) values of 3.89 and 7.19 mg/ml, respectively, compared to two other EOs (C. martinii and O. basilicum). The results suggest that EOs from the genus Cymbopogon can be interesting candidates for nematode control in cattle, although it may prove challenging to deliver concentrations to the gastrointestinal tract sufficient to effectively manage GINs.

Third-stage larvae of the anisakid nematode Contracaecum osculatum infecting cod (Gadus morhua) liver elicit a host immune response involving both innate and adaptive factors, but the reactions differ between liver and spleen. Inflammatory reactions occur in both liver and spleen, but a series of immune effector genes are downregulated in liver infected with nematodes whereas these genes in spleen from the same fish are upregulated. A series of novel primer and probe sets targeting cod immune responses were developed and applied in a real-time quantitative polymerase chain reaction set-up to measure the expression of immune-relevant genes in liver and spleen of infected and uninfected cod. In infected liver, 12 of 23 genes were regulated. Genes encoding cytokines associated with inflammatory reactions (IL-1β, IL-6, IL-8) were significantly upregulated, whereas genes encoding effector molecules, assisting the elimination of pathogens, C-reactive protein (CRP)-PII, hepcidin, lysozyme G1, lysozyme G2, C3 and IgDm, were significantly downregulated. The number of downregulated genes increased with the parasite burden. In spleen, 14 of 23 immune genes showed significant regulation and nine of these were upregulated, including genes encoding CRPI, CRPII, C3, hepcidin and transferrin. The general gene expression level was higher in spleen compared to liver, and although inflammation was induced in nematode-infected liver, the effector molecule genes were depressed, which suggests a worm-induced immune suppression locally in the liver.

Aim of the study was to get in-depth information on Aelurostrongylus abstrusus first-stage larvae (L1s) survival at different temperatures and to assess the capability of these larvae to develop into the third infective stage (L3s). Faeces of a naturally infected cat were split into two aliquots: the first was divided in subsamples assigned to four groups (F1–F4); from the second aliquot, L1s were extracted by Baermann technique, suspended in water and divided into four groups (W1–W4). Groups were stored at different temperatures (F1/W1 −20 ± 1°C; F2/W2 +4 ± 1°C; F3/W3 +14 ± 1°C; and F4/W4 +28 ± 1°C) and L1s vitality assessed every seven days. The capability of L1s stored in water to develop into L3s in snails was evaluated at the beginning and every 21 days. The L1s of W2 and F2 groups remained viable for a longer period (231 and 56 days, respectively) compared to those of other groups. The capability of L1s to moult into L3s in snails showed a decreasing trend; the group W2's L1s maintained the capability to moult into L3s for the longest time (day 189) compared to the other groups. The time of survival of A. abstrusus L1s is influenced by temperature. However, the species seems to be more resistant to temperature variations than other feline lungworms, and this may explain its wider distribution across Europe.

The gastrointestinal helminth parasites of 170 common wallaroos or euros, Osphranter robustus (Gould), collected from all mainland states in which the species occurs as well as the Northern Territory, are presented, including previously published data. A total of 65 species of helminths were encountered, including four species of anoplocephalid cestodes found in the bile ducts and small intestine, and 61 species of strongylid nematodes, all but two of which occurring in the stomach, and with the remainder occurring in the terminal ileum, caecum and colon. Among the mainland subspecies of O. robustus, 52 species of helminths were encountered in O. r. robustus, compared with 30 species in O. r. woodwardi and 35 species in O. r. erubescens. Of the parasite species encountered, only 17 were specific to O. robustus, the remaining being shared with sympatric host species. Host-specific species or species occurring in O. robustus at a high prevalence can be classified as follows: widely distributed; restricted to northern Australia; restricted to the northern wallaroo, O. r. woodwardi; found only in the euro, O. r. erubescens; found essentially along the eastern coast of Australia, primarily in O. r. robustus; and species with highly limited regional distributions. The data currently available suggest that the acquisition of a significant number of parasites is due to co-grazing with other macropodids, while subspeciation in wallaroos as well as climatic variables may have influenced the diversification of the parasite fauna.

Drug resistance to helminth parasites is one of the most serious problems to threaten the livestock industry. The problem also poses a major threat to public health. Therefore, novel and safe agents should urgently be investigated to control parasitic infections. The current study was conducted to evaluate the possible antiparasitic effects of zinc oxide nanoparticles (ZnO-NPs) on one of the most prevalent gastrointestinal nematodes, Teladorsagia circumcincta. The worms were incubated with various concentrations of ZnO-NPs: 1, 4, 8, 12 and 16 ppm for 24 hours. Mobility and mortality of the parasites were recorded at four-hour intervals. At the endpoint, several biomarkers of oxidative/nitrosative stress, including superoxide dismutase, glutathione peroxidase and catalase, as well as lipid peroxidation, protein carbonylation, total antioxidant status, nitric oxide contents and DNA damage, were measured in the homogenized samples. ZnO-NPs showed significant anthelminthic effects, depending on time and concentration. Furthermore, the nanoparticle induced severe oxidative/nitrosative stress and DNA damage. ZnO-NPs could be considered as a novel and potent anthelminthic agent.

The interaction of the rat tapeworm, Hymenolepis diminuta, with its intermediate beetle host, Tribolium confusum, is a well-studied model system. However, there is so much variation in the methods and designs of the studies that it is difficult to draw comparisons. This study simultaneously compared several aspects of the infection protocol including beetle age, sex, density and mating status; parasite egg condition, infection environment humidity and the times for the three steps of infection: starvation, feeding and post-feeding development. Although statistically limited by low prevalence and intensity levels, we were able to detect the relative effects of the variables. The effects of these variables on prevalence (percent infection) and intensity (mean number of cysticercoids) do not necessarily correlate with each other. Egg condition, reduced starvation times, higher beetle density and longer development times reduced prevalence. However, differences in intensity were only detected with older beetles. When coupled with survivorship data, our study found that our current infection protocol is optimal for infection success. However, the results suggest extending the study to other intermediate hosts and the inclusion of additional variables.

Metacercariae of various species within the genus Holostephanus Szidat, 1936 (Trematoda: Digenea: Cyathocotylidae) occur in muscles of both farmed and wild fish, including common carp (Cyprinus carpio Linnaeus, 1758). The life cycle includes a snail as first intermediate host, fish as second intermediate host and birds or mammals as final hosts. We studied the zoonotic potential and the viability of Holostephanus metacercariae from common carp following exposure to various physical and chemical treatments. Muscle tissue samples of common carp specimens from a fish farm in the north-eastern part of Hungary were examined and metacercariae recovered. The zoonotic potential was evaluated experimentally by using small mammals as models (albino mice, n = 2; and Syrian hamsters, n = 4) infected per os with Holostephanus cysts. Parallelly, Metagonimus metacercariae were used as positive controls. We could not confirm the zoonotic potential of Holostephanus metacercariae as they did not survive in the mammalian intestine whereas Metagonimus metacercariae developed to the adult stage. We assessed the viability of metacercariae isolated from common carp specimens during exposure to different physical treatments (temperatures of −18°C, +20°C, +40°C and +60°C) and chemical agents (5% and 10% acetic acid and 10% sodium chloride (NaCl)). Metacercariae lost viability by freezing at −18°C (2 h), heating at 60°C (20 min), incubation in 5% and 10% acetic acid (5 min) and 10% NaCl (2 h). These methods served as models to investigate the effectiveness of food preparation techniques (such as cold and hot smoking, freezing, salting and pickling) on the survival of metacercariae.

Glutathione S-transferases (GSTs) are a detoxifying enzyme family that is essential for parasite blood-feeding and survival, and represent potential targets for hookworm vaccine development. Multiple GST-encoding complementary DNAs (cDNAs) have been cloned from Ancylostoma caninum and Necator americanus, but there are no reports about the cloning of this enzyme from Ancylostoma ceylanicum, the animal-derived zoonotic hookworm. To study the molecular nature and tissue localization of GST of A. ceylanicum (Ace-GST), we designed primers based on the GST gene sequence of A. ceylanicum in GenBank, amplified the Ace-GST cDNA by reverse transcription polymerase chain reaction, and analysed its homology and genetic evolution relationship. The amplified product was cloned into the pET-32a vector and transformed into Escherichia coli BL21 (DE3) for expression. To prepare anti-GST polyclonal antibodies, the recombinant protein was purified and used to immunize Kunming mice. The level of immunoglobulin G (IgG) antibody in the serum of immunized mice was detected by indirect enzyme-linked immunosorbent assay, and the Ace-GST localization in adult worm was determined using the immunofluorescence method. The results showed that the full-length cDNA encoding Ace-GST was 468 bp, which had the highest homology with Ac-GST-1 (60.1%) and clustered into one branch (v-class) with Ac-GST-1 and Na-GST-1 in a phylogenetic tree. Mice immunized with recombinant Ace-GST showed specific IgG antibody response. Immunolocalization revealed that natural Ace-GST is mainly located in the epidermis, muscle and intestine of the adult. These results may lay a foundation for further studies on the biological function of Ace-GST.

Pseudoacanthocephalus goodmani n. sp. is described from faecal pellets collected from Sclerophrys gutturalis (Power, 1927), the guttural toad. The species is characterized by a suite of characters, including a proboscis armature of 14–18 longitudinal rows of 4–6 hooks with simple roots, lemnisci longer than the proboscis receptacle, equatorial testes, a cluster of elongated cement glands and eggs without polar prolongations of the middle membrane 72.6–85.8 long. The toad had been accidentally translocated from Mauritius to the UK in a tourist's luggage and survived a washing machine cycle. The guttural toad was introduced into Mauritius from South Africa in 1922 and the cane toad, Rhinella marina (Linneaus, 1758), from South America, between 1936 and 1938. It seems most likely, therefore, that P. goodmani was introduced, with the guttural toad, from South Africa. The cane toad is host to the similar species, Pseudoacanthocephalus lutzi, from the Americas, but P. lutzi has not been recorded from places where the cane toad has been introduced elsewhere. Clearly, the guttural toad is a hardy and adaptable species, although it seems unlikely that it could become established in Northern Europe. Nevertheless, any accidental translocation of hosts poses the potential risk of introducing unwanted pathogens into the environment and should be guarded against.

Bithynids snails are a widespread group of molluscs in European freshwater systems. However, not much information is available on trematode communities from molluscs of this family. Here, we investigate the trematode diversity of Bithynia tentaculata, based on molecular and morphological data. A total of 682 snails from the rivers Lippe and Rhine in North Rhine-Westphalia, Germany, and 121 B. tentaculata from Curonian Lagoon, Lithuania were screened for infections with digeneans. In total, B. tentaculata showed a trematode prevalence of 12.9% and 14%, respectively. The phylogenetic analyses based on 55 novel sequences for 36 isolates demonstrated a high diversity of digeneans. Analyses of the molecular and morphological data revealed a species-rich trematode fauna, comprising 20 species, belonging to ten families. Interestingly, the larval trematode community of B. tentaculata shows little overlap with the well-studied trematode fauna of lymnaeids and planorbids, and some of the detected species (Echinochasmus beleocephalus and E. coaxatus) constitute first records for B. tentaculata in Central Europe. Our study revealed an abundant, diverse and distinct trematode fauna in B. tentaculata, which highlights the need for further research on this so far understudied host–parasite system. Therefore, we might currently be underestimating the ecological roles of several parasite communities of non-pulmonate snail host families in European fresh waters.