Larval stages of pentastomids were collected from different organs of small mammals from the Peruvian Amazon. These parasitized mammals included: a western Amazonian oryzomys (Hylaeamys perenensis), an elegant oryzomys (Euryoryzomys nitidus), a lowland paca (Cuniculus paca), two kinkajous (Potos flavus), two silvery woolly monkeys (Lagothrix poeppigii) and a brown-mantled tamarin (Leontocebus fuscicollis). Pentastomids were found in the mesentery and parenchyma of the liver and lungs of these animals. All pentastomids were morphologically identified as nymphs of Porocephalus spp. Only the nymphs collected from select animals (the western Amazonian oryzomys, the elegant oryzomys and the brown-mantled tamarin) were analysed molecularly. Molecular analysis was performed amplifying the mitochondrial cytochrome c oxidase subunit I gene from select nymphs collected from the western Amazonian oryzomys, the elegant oryzomys and the brown-mantled tamarin. The nucleotide sequences exhibited 95.8–97.7% similarity between them. Also, these sequences showed an identity of 95.8–97.9% to Porocephalus crotali (GenBank accession numbers MG559647–MG559655). Molecular analysis indicated the presence of at least two Porocephalus species. These findings represent the first record of Porocephalus in these mammals, thus adding new intermediate hosts for this pentastomid genus. This work represents the first molecular data of Porocephalus in a Neotropical climate.

Whipworm infection or trichuriasis caused by Trichuris trichiura is of major public health concern in sub-Saharan Africa, particularly among pre-school and school-going children. It is among the neglected tropical diseases targeted for elimination through mass drug administration (MDA). One of the outcomes of MDA is a rapid decline in levels of infection intensity, making it difficult to monitor effectiveness of control measures using the conventional Kato–Katz procedure, which relies on the microscopic detection of parasite ova in faecal samples. In the present study, a loop-mediated isothermal amplification (LAMP) test was developed for the detection of T. trichiura infection in faecal samples. LAMP technology offers greater sensitivity and specificity than the microscopy-based tests. A set of four specific primers targeting the internal transcribed spacer 2 region of the ribosomal DNA were designed using Primer Explorer software. DNA was extracted from faecal samples using the alkaline lysis method (HotSHOT) and the LAMP reaction performed at 63°C for 1 h. The amplicons were visualized by both gel electrophoresis and with the naked eye following staining with SYBR green dye. Sensitivity and specificity tests were determined using the standard Kato–Katz diagnostic procedure as a reference test. The developed LAMP assay reliably detected T. trichiura DNA in faecal samples, with a specificity and sensitivity of 88% and 77%, respectively. No cross-reactivity was observed with several common helminth parasites. The developed LAMP assay is an appropriate diagnostic method for the detection of T. trichiura DNA in human faecal samples due to its simplicity, low cost, high sensitivity and specificity.

Parasites can have strong effects on invertebrate host behaviour, fecundity and survival in marine ecosystems. However, parasites are often poorly documented and still rarely integrated into marine ecological modelling; comprehensive surveys of infection in marine invertebrates are sporadic at best. For example, rock crabs are an important part of Californian coastal ecosystems, both as regulators of mussel populations and non-native species, and as prey items for predators like sea otters, but their parasite communities and potential effects on crab population dynamics are seldom studied or understood. Here, we present the first report of infection by the trematode Helicometrina nimia in the economically and ecologically important red rock crab (Cancer productus) and Pacific rock crab (Romaleon antennarium). As intermediate hosts, they are a missing link for infection by H. nimia in Californian fish that was unreported until now. Based on these findings, we advocate for further research into parasite diversity and their potential effects on ecologically and commercially important species.

Here, we report the first record of pleroceroids of the Nearctic tapeworm Proteocephalus ambloplitis (Leidy, 1887) in European fresh waters. The plerocercoids were found encapsulated in the internal organs of the invasive pumpkinseed Lepomis gibbosus (Linnaeus, 1758) in a pond in Bègles Plage, Bordeaux, France. This parasite species was not observed in L. gibbosus collected from two nearby localities, Bordeaux Lac and a pond in the Parc de Fontaudin. Species identification was confirmed using molecular data and morphological characteristics.

Epidemiological studies of Echinococcus multilocularis infections in definitive hosts require a reliable and economic diagnostic method. In this study, the current copro-DNA examination technique was modified by increasing the faecal amounts tested and adding a step to neutralize the faeces before DNA extraction. Reliability of the modified method was evaluated using rectal faecal samples from red foxes and comparing them with intestinal worms detected using the sedimentation and counting technique (SCT) following necropsy. The modified copro-DNA examination method demonstrated 93.9% sensitivity (138/147) on the SCT. Its detectability increased depending on the worm burden, and the sensitivity was 100% in cases harbouring over 1000 worms. From 111 SCT-negative cases, six (5.4%) were copro-DNA-positive, and all were confirmed as E. multilocularis via sequencing analysis. Five of the remaining 105 SCT-negative cases (4.8%) retained polymerase chain reaction (PCR) inhibitors in the extracted solution, suggesting that approximately 5% of the red fox faeces retained these inhibitors after treatment with the present copro-DNA extraction method. Although further evaluation is needed for faeces deposited in the wild, the present copro-DNA examination technique will help monitor the E. multilocularis prevalence in definitive hosts. When used for detailed evaluations of endemicity (e.g. changes in infection pressure or spread in non-endemic areas), the absence of PCR inhibitors should be confirmed, and multiple trials on faecal subsamples are recommended.

Knowledge of the parasite fauna of Australian freshwater fish is fragmentary and incomplete. An understanding of fish hosts and their associated parasites is vital for the successful management of aquatic ecosystems. In this study, we surveyed the parasite fauna of carp gudgeons (Hypseleotris spp.), a complex of species of Australian freshwater fishes, using morphology and molecular data for the 18S and 28S ribosomal RNA genes. We examined 137 individuals of three different taxa in the carp gudgeon species complex and found 16 parasitic taxa of the Digenea, Cestoda, Nematoda and Arthropoda (five adults and 11 larvae). Eleven parasites are reported for the first time from the carp gudgeons (Pseudodactylogyrus sp., Gyrodactylus sp., Clinostomum sp., Paradilepis patriciae, P. cf. kempi, two unidentified species of Paradilepis, Dendrouterina sp., Parvitaenia sp., two lineages of Cyclophyllidea gen. sp., Procamallanus sp., larvae of a spirurine nematode and Lernaea sp.), in addition to Apatemon cf. hypseleotris Negm-Eldin & Davies, 2001 and the invasive tapeworm Schyzocotyle acheilognathi (Yamaguti, 1934), which were previously reported from these fish hosts. Parasite species richness was double in Lake's and Midgley's carp gudgeons relative to western carp gudgeon. These findings highlight the key role of carp gudgeons as intermediate hosts for multiple parasites with complex life cycles using native birds as definitive hosts and the usefulness of DNA data for the identification of parasite larvae.

The lack of anthelmintic products licensed for donkeys and the rising number of small donkey milk farms in the countries of Western Europe and Italy have led to an increased interest in the study of reliable and safe plant-derived treatment alternatives. In this study, the aqueous extracts of Achillea millefolium L. (flowers), Artemisia absinthium L. (aerial parts), Centaurium erythraea Rafn. (flowers), Gentiana asclepiadea L. (rhizomes and roots), Inula helenium L. (rhizomes and roots) and Tanacetum vulgare L. (aerial parts), have been tested in vitro for their potential ovicidal and larvicidal activity against donkey nematodes. An egg-hatching assay (EHA) and larval development assay (LDA) were performed for the in vitro study, and median lethal concentration (LC-50) values for both EHA and LDA were calculated using probit analysis. All tested plant extracts showed strong anthelmintic activity against strongyle eggs and larvae at concentrations ranging between 125 and 1.95 mg/ml, except for C. erythraea, which exhibited very little or no effect at all at the tested concentrations. A strong ovicidal effect was observed in A. absinthium, with an LC-50 value of 0.486 mg/ml (95% confidence interval (CI) 0.21–1.09). Gentiana asclepiadea showed high efficacy against strongyle larvae, with an LC-50 value of 0.041 mg/ml (95% CI 0.01–0.16). The most significant (P < 0.01) anthelmintic activity was exhibited by I. helenium, with an LC-50 value of 0.041 mg/ml (95% CI 0.01–0.16) for EHA and 0.41 mg/ml (95% CI 0.27–0.62) for LDA. The results proved the anthelmintic efficacy of the tested plant extracts, highlighting the need for further research into plant bioactive molecules both in vitro and in vivo.

In the Laboratory of Parasites of Fishes, Crustaceans and Mollusks (CEPAVE), we undertook a parasitological study on three species of fish from the Espinal and Esteros del Iberá ecoregions of Argentina. Clinostomid metacercariae were found parasitizing Characidium rachovii, Crenicichla vittata and Gymnogeophagus balzanii. In this study, we analysed the damage that these parasites inflict on their hosts through the evaluation of histological sections. In addition, Clinostomum metacercariae were identified using morphological characters and DNA barcoding. In the pathological analysis, we observed that muscle tissue was the most affected. The inflammatory response showed vascular congestion areas and infiltration of numerous inflammatory cells, mainly lymphocytes. The molecular and morphological approach supports the presence of three new lineages of clinostomid metacercariae in Argentina. This could lead to the discovery of a high number of lineages or species of Clinostomum from South America.

Thada populus n. sp. was found in the rhizosphere of Populus euphratica in the city of Dezful, south-western Iran. The new species is characterized by its 365–453 μm body length, cuticle with transverse annuli 0.9–1.2 μm wide, lateral fields with four lines, lip region low with one or two annuli, 7.0–8.5 μm wide at base, longitudinal or slightly sigmoid amphidial slit, delicate stylet 8.4–9.8 μm long with posteriorly directed knobs, dorsal pharyngeal gland opening at 1.0–1.5 μm posterior to the stylet knobs, non-muscular and valve-less median bulb, pyriform and offset basal bulb, almost oval spermatheca, posterior position of vulva (V = 75–79%) without lateral membrane, short post-vulval uterine sac and conical tail with finely to broadly rounded terminus. Morphological differences of the new species with those of the type species, Thada striata, are discussed. Molecular phylogenetic studies of the new species using partial sequences of small subunit ribosomal DNA revealed that the new species formed a clade with Malenchus spp., Filenchus facultativus, F. fungivorus and Filenchus sp. in Bayesian inference. Morphological and molecular phylogenetic studies were also performed on Tenunemellus indicus, the second recovered species, the latter analysis using partial sequences of large subunit ribosomal DNA. In the inferred Bayesian tree, T. indicus formed a clade with Labrys fuzhouensis, Lelenchus leptosoma from the Netherlands, Malenchus spp. and Filenchus discrepans.

Many temperate freshwater habitats are at risk for contamination by run-off associated with the application of road de-icing salts. Elevated salinity can have various detrimental effects on freshwater organisms, including greater susceptibility to infection by parasites and pathogens. However, to better understand the net effects of road salt exposure on host–parasite dynamics, it is necessary to consider the impacts on free-living parasite infectious stages, such as the motile aquatic cercariae of trematodes. Here, we examined the longevity and activity of cercariae from four different freshwater trematodes (Ribeiroia ondatrae, Echinostoma sp., Cephalogonimus sp. and an unidentified strigeid-type) that were exposed to road salt at five different environmentally relevant concentrations (160, 360, 560, 760 and 960 mg/ml of sodium chloride). Exposure to road salt had minimal detrimental effects, with cercariae activity and survival often greatest at intermediate concentrations. Only the cercariae of Cephalogonimus sp. showed reduced longevity at the highest salt concentration, with those of both R. ondatrae and the unidentified strigeid-type exhibiting diminished activity, indicating interspecific variation in response. Importantly, cercariae seem to be relatively unaffected by salt concentrations known to increase infection susceptibility in some of their hosts. More studies will be needed to examine this potential dichotomy in road salt effects between hosts and trematodes, including influences on parasite infectivity.

Adult forms of members of the Callodistomidae always parasitize the gallbladder of freshwater fishes and occur in Africa and America. This study provides a description of a new South American species belonging in Prosthenhystera from the gallbladder of a characid fish (Bryconamericus ikaa), and ribosomal gene sequences (28S rDNA and ITS1-5.8S-ITS2) are used to demonstrate molecular differences between the new species and congeners as well as explore interrelationships among congeners. Additionally, the first cytological analysis is conducted for a member of the family to determine chromosome number and arrangement. Prosthenhystera gattii n. sp. most closely resembles Prosthenhystera caballeroi in morphology, but the vitellarium is more extensive reaching anterior to the caecal bifurcation in the new species and the uterus is confined to the hindbody in P. gattii n. sp., whereas it extends to the level of the pharynx in P. caballeroi. Also, the testes, cirrus sac, seminal receptacle and the ratio of body length to width are larger in P. gattii n. sp. Independent Bayesian inference analyses of 28S rDNA and ITS1-5.8S-ITS2 sequence fragments produced phylograms that showed P. gattii n. sp. is more similar to Prosthenhystera obesa + Prosthenhystera oonastica than P. caballeroi + two unidentified species of Prosthenhystera, but with poor posterior probability support for the node in the ITS1-5.8S-ITS2-based phylogram. Further, the genetic distance between P. oonastica and P. gattii n. sp. are the largest among Prosthenhystera spp. Cytological analysis revealed ten metacentric chromosomes, which is fewer than the 12–18 chromosomes present in species from the closely related Gorgoderidae.

Comparative analysis using complete ITS2 and partial 28S rDNA sequence data revealed that cercariaeum developing in rediae in Lithoglyphus naticoides represent two different lissorchiid species. One morphotype of cercariaeum is conspecific with adult Palaeorchis incognitus from European roach, Rutilus rutilus. The other cercariaeum is attributable to the genus Asymphylodora, but the species identity is not yet determined. We also generate the first rDNA sequences for Asymphylodora progenetica based on new collections from Bithynia tentaculata from Lithuania. Phylogenetic analyses of the newly generated sequences, together with information for other lissorchiids available on GenBank, showed that all representatives of Lissorchiidae form a strongly supported clade. Three monophyletic lineages, Asymphylodora, Palaeorchis and Lissorchis, were recognized at the generic level. Karyological analysis of the chromosome set of larval P. incognitus revealed a diploid number of 2n = 20. Its karyotype with subtelocentric chromosomes prevailing can be regarded as comparatively ‘primitive’, which is consistent with the basal position of P. incognitus in the 28S tree relative to the representatives of the genus Asymphylodora. The present study adds significant new information for establishing species-specific markers for the confident characterization of different developmental stages of lissorchiid species and clarification of their life cycles.

Helminthological studies may contribute with valuable information on host biology and conservation. Herein, we provide new data on helminths infecting the lizard Norops fuscoauratus, testing one of the factors considered most important in parasitic ecology: host size. We analysed 25 specimens of N. fuscoauratus from three highland marshes in the Brazilian semi-arid. Eight taxa of helminths belonging to Nematoda, Trematoda and Acanthocephala were found. Physaloptera sp. showed the higher prevalence (40%), with a mean intensity of infection of 3.3 ± 1.46 (1–16) and mean abundance 1.32 ± 0.65 (0–16). Norops fuscoauratus represents four new host records for the helminths Cyrtosomum sp., Pharyngodon travassosi, Strongyloides sp. and Centrorhynchus sp. There is no relationship of host body size (P = 0.79) and mass (P = 0.50) with parasite richness. In addition, the present study contributes to the knowledge of the parasitic fauna of N. fuscoauratus and the Neotropical region.

A description and the molecular characterization of two new species in the Haploporidae and Haplosplanchnidae families are provided herein. Parasaccocoelium armatum n. sp. was collected from the intestine of a Mugil cephalus Linnaeus, 1758 from the Primorsky region, Russia, and Pseudohaplosplanchnus catbaensis n. g. n. sp. was collected from Moolgarda seheli (Forsskål, 1775) in the coastal waters of Cat Ba Island, Vietnam. The morphological features of P. armatum n. sp. closely resemble those of Parasaccocoelium polyovum, but these species differ from one another by hermaphroditic sac and vitellaria area length and by maximal egg size. The main difference between P. armatum n. sp. and P. polyovum is the presence of an armed hermaphroditic duct in the new species. Molecular data support the case for inclusion of the studied trematodes in P. armatum n. sp. Worms P. catbaensis n. g. n. sp. from the mullet from Vietnam are morphologically close to Haplosplanchnus (Haplosplosplanchninae). The only difference between P. catbaensis n. g. n. sp. and species of Haplosplanchnus is the presence of few (1–7) large eggs, measuring 135–142 × 92–104 μm, versus numerous small eggs with a maximal size of 75 × 50 μm. Phylogenetic analysis showed that there is a contradiction between the morphological similarity of the worms and their position in the Haplosplanchnidae system, based on the genetic data. Results of this study indicate that P. catbaensis n. g. n. sp. is genetically distant from other representatives of Haplosplanchnus, despite their morphological similarity. According to the molecular data, P. catbaensis n. g. n. sp. is close to Hymenocotta mulli Manter, 1961 (Hymenocottinae). However, these species are considerably different to each other morphologically. Molecular data argue for the possibility of establishing a new subfamily for P. catbaensis n. g. n. sp. However, considering earlier studies of Haplosplanchnidae, we support the view that creating new subfamilies within this family is unreasonable because of the lack of molecular data for most haplosplanchnid species, which are necessary to resolve the problematic systematics and phylogeny of this family.

We suggest helminthological investigations of cod as a supplement to traditional biological and hydrographical methods for elucidation of ecological changes in the Baltic Sea. It is under discussion if oxygen deficit or seal abundance should explain the present critical situation of Baltic cod. A comparative investigation of endoparasitic helminths in Baltic cod (Gadus morhua), captured in the same marine habitat with an interval of 35 years (1983/2018) recorded 11 species of helminths comprising trematodes (Hemiurus luehei, Podocotyle atomon, Lepidapedon elongatum), nematodes (Contracaecum osculatum, Hysterothylacium aduncum, Capillaria gracilis, Cucullanus cirratus), cestodes (Bothriocephalus sp.) and acanthocephalans (Echinorhynchus gadi, Pomphorhynchus laevis, Corynosoma semerme). Significant prevalence and intensity increases were recorded for third-stage larvae of the nematode C. osculatum (liver location) and larvae of C. semerme (encapsulated in viscera). Both parasite species use grey seal as their final host, indicating the recent expansion of the Baltic seal population. A lower E. gadi intensity and an increased prevalence of L. elongatum of small cod (31–40 cm body length) suggest a lowered intake of amphipods (intermediate host) and elevated ingestion of polychaetes, respectively, but no significant changes were seen for other helminths.

A new species of the genus Lyperosomum Looss, 1899, from the intestine of the golden-fronted woodpecker (Melanerpes aurifrons) from northern Mexico is described. Lyperosomum cuauhxinqui sp. n. is morphologically distinguished from other congeneric species from the Americas by a higher oral/ventral sucker ratio and its body length and width. The sequences of domains D1–D3 of the large subunit (LSU) of nuclear ribosomal DNA and cytochrome c oxidase subunit 1 (cox 1) from the mitochondrial DNA of the new species were obtained and compared with available sequences from GenBank. The genetic divergence estimated between the new species and other congeneric species ranged from 2 to 6% and 13.4 to 17.3% for LSU and cox 1, respectively. Phylogenetic analyses based on the two (LSU and cox 1) molecular markers consistently showed that L. cuauhxinqui sp. n. was nested within the genus Lyperosomum, with strong bootstrap support (100%) and Bayesian posterior probabilities (1.0). In particular, the LSU tree indicated that the sequence of the new species is closely related to sequences from Zonorchis alveyi, Zonorchis delectans and Zonorchis sp. from Central America, suggesting that these sequences should be transferred to the genus Lyperosomum. The new species represents the first record from Mexico and the fifth species identified in the Americas. Our study also revealed that the taxonomy of the genus Lyperosomum should be re-examined by combining molecular, morphological and ecological characteristics.

Echinococcus granulosus is the causative agent of cystic echinococcosis, which has serious impacts on human and/or animal health, resulting in significant economic losses. Echinococcus granulosus comprises a number of intra-specific variants or strains at the genetic level. In Saudi Arabia, few studies were performed on genetic variations in Echinococcus species. Therefore, the present study aimed to investigate the phenotypic and genetic characterization of hydatid cysts harboured by sheep and camels in Al-Madinah Al-Munawarah. Samples of hydatid cysts were collected from local sheep (n = 25) and camels (n = 8). The morphological criteria of protoscoleces were investigated. To investigate the molecular characterization, random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR), single-stranded conformation polymorphism (SSCP) were carried out. DNA was extracted from individual fertile cysts and subjected to RAPD-PCR analysis (using five arbitrary primers) and PCR amplification of cytochrome c oxidase I (cox1) and 12S ribosomal ribonucleic acid (12S rRNA) genes. The PCR products were subjected to SSCP analysis for genetic discrimination in E. granulosus isolates. In addition, partially sequencing of the mitochondrial DNA cox1 genes was achieved for assessing the phylogenetic positions of collected isolates using some global published sequence data of cox1 genes. The rostellar hooks of camel and local sheep isolates show remarkable variability in their dimensions. Five distinct SSCP patterns were identified in the 12S rRNA gene, showing intraspecific variations in E. granulosus of camels and local sheep. Sequencing of (cox1) genes of both local sheep and camels exhibit high similarity with those of the same gene (E. granulosus sensu stricto) published in NCBI BLAST.

Few digeneans of the family Fellodistomidae are known from the Russian Arctic seas. The taxonomic status of these species, their life cycles and host range raised recurrent questions, some of which remain unanswered. To revise the species composition and life cycles of fellodistomids in the White Sea, we searched for them in several known and suspected hosts: wolffish, flatfishes (definitive), gastropods of the family Buccinidae (second intermediate) and protobranch bivalves (first intermediate). Species identification was based both on morphology and 28S ribosomal RNA gene sequences. We found Fellodistomum agnotum in the White Sea for the first time. Buccinum undatum was proved to be intermediate host of both F. agnotum and Fellodistomum fellis, and metacercariae of F. fellis were registered from two more buccinid species: Buccinum scalariforme and Neptunea despecta. We also found metacercariae of F. agnotum and F. fellis producing eggs in the second intermediate host. Two fellodistomids were found in protobranch bivalves: sporocysts and cercariae of Steringophorus furciger in Nuculana pernula, and sporocysts with large furcocercous cercariae in Ennucula tenuis. The latter were identified as F. agnotum by molecular analysis; thus, the entire life cycle of this species was reconstructed.

Because of their continuing expansion, wildlife ruminant species that prosper in rural landscapes may be increasingly affected by and/or contribute to the circulation of certain generalist pathogens also infecting domestic ruminants, when they share common spaces or resources. In this study, we aimed to test the hypothesis that parasitism with gastrointestinal nematodes (GINs) of wild roe deer inhabiting different rural landscapes is correlated with livestock density. We used faecal egg counts of GINs and spatial data of 74 GPS-collared roe deer, inhabiting various landscapes from closed forests to open fields, together with weekly records of livestock abundances on pasture. We tested whether the excretion of GIN eggs in roe deer was influenced by the density of livestock in their home range over the grazing season. Our results showed that all of the roe deer home ranges, except four, contained pastures occupied by livestock. Excretion of GIN eggs occurred in 77% of the roe deer. The excretion of GIN eggs in roe deer tended to increase with livestock density in their home range. This result suggests, but does not prove, a higher risk of ingesting GIN larvae originating from livestock dung. In the context of increasing overlap between roe deer and livestock ranges, the exchange of pathogens between both hosts is plausible, although species identity of the parasites present was not determined. Assessing which GIN species are shared between wild and domestic ruminants, and how this may affect the health of both hosts, is a central question for future research in the context of interspecific pathogen circulation.

The aim of this study was to evaluate the combined use of different chemical (albendazole, ivermectin, glycerine and Vaseline) and biological (Monacrosporium thaumasium) compounds in the control of Ancylostoma caninum. Infective larvae of A. caninum were obtained from coprocultures of positive faeces from naturally infected dogs. We used 1% ivermectin, 1% albendazole, 100% glycerine, 100% Vaseline and an isolate of the nematophagous fungus M. thaumasium (NF34), alone or in combinations. Next, an experimental test was set up with 16 groups in microtubes, with a 24-h interaction. The groups (G1 to G15) that contained any chemical or biological compound (NF34) and/or their combined use (chemical + biological) showed a difference in relation to the control group, except G5 – Vaseline 100% without combinations. It was concluded that, even on an experimental basis, the combined use of anthelmintic drugs with biological control was efficient; however, more studies must be carried out in order to elucidate the synergistic action between chemical and biological compounds to be used in the effective control of hookworms in the future.