In Argentina, the family Diplostomidae is composed of eight genera: Austrodiplostomum Szidat & Nani; Diplostomum von Nordmann; Dolichorchis Dubois; Hysteromorpha Lutz; Neodiplostomum Railliet; Posthodiplostomum Dubois; Sphincterodiplostomum Dubois; and Tylodelphys Diesing. During a parasitological survey of fishes from the Iguazú National Park we detected diplostomid metacercariae in the brain of Erythrinus cf. erythrinus. Fish were caught using crab traps, transported alive to the field laboratory, cold-anaesthetized and euthanized by cervical dissection. Some metacercariae were heat-killed in water and fixed in 10% formalin and others were preserved in alcohol 96% for DNA extraction. They were sequenced for the partial segment of the 28S rDNA, internal transcribed spacer (ITS) rDNA and cytochrome c oxidase subunit I (COI) mtDNA genes. Phylogenetic reconstruction was carried out using Bayesian inference and the proportion (p) of absolute nucleotide sites (p-distance) was obtained. In the 28S rDNA tree, the metacercaria sequenced grouped as Dolichorchis sp. The COI mtDNA p-distance between the metacercariae with Dolichorchis lacombeensis was 0.01. There is a small number of ITS sequences for the Diplostomidae family deposited in the GenBank. The oral sucker, ventral sucker, holdfast organ and the distance between oral and ventral suckers are larger in the adult compared with the metacercariae. Additionally, hind-body length and width are larger in the adult due to the development of the genital complex. Further studies using an integrative approach will help confirm the affiliation of other species to the genus Dolichorchis.

In the present study, we aimed to describe the composition of endoparasites associated with anurans from an altitudinal rainforest enclave in northeastern Brazil. Additionally, we tested if microhabitat use influences endoparasite abundance and richness, as well as the hypothesis that larger frogs tend to be more parasitized. We sampled 306 individuals from 25 anuran species that were necropsied and analysed using a stereomicroscope. The total endoparasite prevalence was 79.08%, with a parasitic community consisting of 46 taxa. Overall, we found the common pattern described for Neotropical amphibians, which is the predominance of generalist and direct-cycle parasites. Twenty new host records and two possible new parasite species were found, highlighting the importance of this type of inventory. We also observed that microhabitat use was associated with a significant difference in parasite richness between groups, in which arboreal and terrestrial species, and aquatic and arboreal species contributed to these differences. Moreover, larger frogs tended to be more parasitized regarding only an interspecific view. Our results suggest that parasite richness is directly related to infection cycle and how the host exploits its habitat.

Rodentolepis (sensu lato) asymmetrica (Janicki, 1904) is redescribed on the basis of materials from Microtus agrestis, Microtus arvalis and Myodes glareolus from Eastern Europe (Belarus, Lithuania, Latvia, Estonia, Ukraine and the north-eastern part of Russia). A new genus, Kontrimavichusia n. g., is proposed for it based on morphological and molecular evidence. The unique morphological diagnostic characters of this currently monotypic genus include the presence of an armature on the suckers, ventral osmoregulatory canals connected with irregularly spaced transverse anastomoses, an internal seminal vesicle with circular musculature and uterus with numerous diverticula, situated dorsally to the genital ducts, extending bilaterally beyond the longitudinal osmoregulatory canals. In addition, the new genus differs from morphologically related genera of the Rodentolepis clade by the structure of its vagina. The copulatory part of the vagina in specimens of Kontrimavichusia n. g. is surrounded by circular musculature and covered externally by a dense layer of intensely-stained cells; the conductive part of the vagina is clearly distinguishable from the seminal receptacle. Molecular analysis of the partial 28S rRNA gene fully supports the erection of Kontrimavichusia n. g. as a distinct lineage.

Cosmocerca spp. are common nematode parasites of amphibians. We provide herein molecular data for two species of Cosmocerca and describe a new species, Cosmocerca albopunctata n. sp., using light microscopy and molecular data (cytochrome c oxidase subunit 1 – COI mtDNA). Cosmocerca albopunctata n. sp. can be easily distinguished from other congeneric species by the combination of characteristics such as body size, length of spicules and gubernaculum, and the arrangements and number of caudal papillae (7 + 1:1 + 1:6). The phylogenetic results based on the partial COI mtDNA sequences clustered the new species in a monophyletic clade along with the other sequences of Cosmocerca spp. Therefore, our results contribute to the knowledge about the species diversity and genetic data for Cosmocerca spp. in the Neotropical region.

In this study, we report the first complete mitochondrial genome of the tapeworm Nippotaenia mogurndae in the order Nippotaeniidea Yamaguti, 1939. This mitogenome, which is 14,307 base pairs (bp) long with an A + T content of 72.2%, consists of 12 protein-coding genes, 22 transfer RNA (tRNA) genes, two rRNA genes, and two non-coding regions. Most tRNAs have a conventional cloverleaf structure, but trnS1 and trnR lack dihydrouridine arms of tRNA. The two largest non-coding regions, NCR1 (220 bp) and NCR2 (817 bp), are located between trnY and trnS2 and between nad5 and trnG, respectively. Phylogenetic analyses of mitogenomic data indicate that N. mogurndae is closely related to tapeworms in the order Cyclophyllidea.

A new species of the genus Paurodontella, Paurodontella minora n. sp., collected from Alborz Province, Iran, is described and illustrated based on morphological and molecular characters. The new species is characterized by its body length of 393 (350–438) μm and 380 (n = 1) μm in female and male, respectively, 6–7 incisures in lateral field, lip region annulated and continuous with body contour, and total stylet 6.1 (5.5–7.0) μm long. Basal pharyngeal bulb with small posterior extension projecting reaching to the intestine. Excretory pore situated at the level of basal pharyngeal bulb region, no post-uterine sac, conical tail, narrowing to a rounded tip, and rare male with slender tylenchoid spicules and adanal bursa. The new species comes close in morphology and morphometrics to four known species of the genus, namely Paurodontella asymmetrica, Paurodontella balochistanica, Paurodontella densa and Paurodontella niger. In molecular phylogenetic analyses using D2–D3 expansion segments of the large subunit rDNA gene sequence, P. minora n. sp. formed a major clade with species of the genera in the family Sphaerulariidae (Paurodontella, Paurodontoides, Veleshkinema and Sphaerularia) and a sister relation with the members in the families Neotylenchidae and Anguinidae with the same clade support values in Bayesian inference.

Genetic markers, DNA sequences and karyotypes, of some European lissorchiid species from their intermediate and final hosts were obtained to clarify controversial data about their life cycles and taxonomy, and to reveal phylogenetic affinities. The life cycles of three species have been confirmed for the first time based on molecular data. Comparative analysis of internal transcribed spacer 2 (ITS2) and partial 28S rDNA sequences has undoubtedly proven that cercariaeum of type-species of the genus Asymphylodora, Asymphylodora tincae, develops in pulmonate snails, Anisus vortex and Stagnicola palustris, but not in the genus Bithynia. The faucet snail, Bithynia tentaculata, serves as the first intermediate host for Parasymphylodora (=Asymphylodora) markewitschi and Parasymphylodora parasquamosa; adults of both species were isolated from the common rudd, Scardinius erythrophthalmus. It has also been confirmed that B. tentaculata serves as the second intermediate host for P. parasquamosa. Phylogenetic analysis supports the validity of the genus Parasymphylodora. Two species, Parasymphylodora markewitschi and P. parasquamosa, with cercariaeum belonging to the Parasquamosum group, are closely related and are being recovered as a well-defined evolutionary lineage in phylogenetic trees. A significant divergence between Parasymphylodora spp. and Asymphylodora spp. was revealed. The diploid chromosome set of P. markewitschi is composed of 14 chromosomes and does not show similarities with karyotypes of other lissorchiid species. Asymphylodora progenetica and Asymphylodora tincae share the basal diploid value of the family, 2n = 20, and reveal very close morphology of the corresponding chromosome pairs. Karyotypic similarities of these species are in accordance with molecular phylogenetic data. Thus, the available molecular and cytogenetic data support the assignment of P. markewitschi and P. parasquamosa to a separate genus, meanwhile, the assignment of A. progenetica to the genus Parasymphylodora was not justified.

Nematoda of the genus Echinocephalus Molin, 1858 include species from the Gnathostomatidae family, the adult stages of which parasitize the stomach and spiral intestine of elasmobranchs as their final hosts. In the present study, we describe Echinocephalus spinosus n. sp. found parasitizing the spiral valve of the freshwater stingray Potamotrygon motoro from the Tapajós River, in the Amazon Basin, in the state of Pará, Brazil. In the study we performed morphological (light and scanning electron microscopy) and molecular (small subunit ribosomal ribonucleic acid (SSU rDNA) and mitochondrial cytochrome c oxidase I sequencing) analyses. E. spinosus n. sp. is only the second species of the Echinocephalus genus described from a strictly freshwater environment. The SSU rDNA based phylogenetic analyses showed Echinocephalus clade as a sister lineage of Gnathostoma, and that the new species arises as a sister to Echinocephalus cf. pseudouncinatus. Time-calibrated phylogenetic analysis revealed that the origin of the freshwater Echinocephalus coincides with the recently proposed origin of the freshwater host potamotrygonin stingray, namely the late Oligocene to early Miocene, when the western Amazon was dominated by the Pebas wetlands, an epicontinental marine/freshwater system.

A new species of the genus Oscheius, Oscheius cyrus n. sp., collected in the moist soils taken from forest heights in the north of Iran, is recorded. A comprehensive description, comprising molecular (internal transcribed spacer (ITS), 18S, and 28S rDNA genes) information, morphometrics data, light microscope and scanning electron microscope images, is supplied. The species resembles Oscheius myriophilus. However, the highest ranges for female body length, female tail, infective juvenile tail length, median bulb, absence of epiptygma and lateral field incisures number vary. The new species was distinguished from Oscheius insectivorus by the general lip region. The male was not found. Molecular analysis showed that the new species has the most similarity to O. myriophilus both in the ITS and 18S regions. Morphological and molecular data confirmed its belonging to the Insectivora-group. Furthermore, the species of Ochrobactrum pseudogrignonense was reported as a dominant associated bacterium of the new Oscheius species. Finally, the mortality of the host after seven days varied from 20% to 82.5%, depending on nematodes’ concentration.

Plerocerci of the trypanorhynch cestode Progrillotia dasyatidis Beveridge, Neifar & Euzet, 2004 (Progrillotiidae) were isolated from the gallbladder of the three-spined stickleback Gasterosteus aculeatus from a small freshwater marsh in the Lake Atanasovsko Wetlands, a coastal area adjacent to the Black Sea coast, Bulgaria. The parasite was recorded in five out of 134 fish individuals studied (prevalence 3.73%, intensity 1–7, mean intensity 2.40 ± 1.17 and mean abundance 0.09 ± 0.06). A description of the plerocerci is presented, expanding data on intraspecific variation. The present report is the first record of P. dasyatidis from G. aculeatus (new host record) and from Bulgaria (new geographical record). Recording only plerocerci with evaginated scoleces in the sticklebacks is consistent with the hypothesis that teleosts are paratenic hosts and not intermediate hosts of P. dasyatidis.

Trichinellosis is an important worldwide foodborne zoonosis. The gold standard test to detect Trichinella spp. larvae in muscle samples of animals intended for human consumption is the artificial digestion method. Handling and dispensing of conventional pepsin powder present significant safety risks for analysts. The use of pepsin powder that is resistant to aerosolization should alleviate these safety concerns. The aim of this study was to compare the efficacy of an aerosol-resistant pepsin powder to conventional pepsin powder in the artificial digestion method. Proficiency samples of pork diaphragm containing specific numbers of viable Trichinella spiralis larvae were tested in two laboratories. The results revealed that aerosol-resistant pepsin was simple, effective and convenient to use, and showed good solubility and larval recovery that met the requirements of the European Union regulation EU 2015/1375. Overall, the efficacy of the aerosol-resistant pepsin was comparable to the conventional pepsin and safer for analysts.

The type species of the genus Epacrolaimus, Epacrolaimus declinatoaculeatus, is studied from the re-examination of type material of Aporcelaimus vorax, its junior synonym, and the observation of several Iberian populations and a few Iranian specimens. Morphologically, it displays a recognizable morphological pattern characterized by, among other features, the incurved nature of its odontostyle aperture, presence of perioral liplets or lobes, lip region 24–31 μm wide, odontostyle 21–25 μm long and comparatively anterior location of S2N pharyngeal gland nuclei. Nevertheless, variations in some morphological traits (vagina shape and tail shape) and in several morphometrics (body length, uterus length, vulva position, tail length and spicule length) are also noted. Sequences of D2–D3 domains of the 28S rDNA, 18S rDNA and COI mtDNA were obtained from several Iberian populations. Their analyses, in particular those from D2–D3 sequences, revealed the existence of a highly supported clade ((Epacrolaimus + Sectonema) + Metaporcelaimus), with a closer relationship between Epacrolaimus and Palaearctic populations of Sectonema, whereas the remaining aporcelaimid genera occupied placements in other clades. These results are discussed, with especial emphasis on the intricate separation of Epacrolaimus and Sectonema, which display significantly different protruding stomatal structure in spite of their close evolutionary relationship as derived from molecular trees.

Parasitic ‘turbellarians’ are known from various animals such as echinoderms, crustaceans, annelids, bivalve and gastropod molluscs. So far, however, no ‘turbellarians’ have been reported from cephalopods. In this paper we report a parasitic ‘turbellarian’ from the giant Antarctic octopus, Megaleledone setebos. We dissected two specimens of M. setebos caught in the Ross Sea (Antarctica) and found numerous worms in their intestine and liver. The worms were spherical or oblong and had two morphologically different poles. The frontal pole bears a small conical protrusion containing large elongated pear-shaped frontal glands and large polygonal cells. The ducts of the frontal glands open terminally to form the frontal organ. The caudal pole has an opening shaped as a folded tube connected by the genital pore with a common genital atrium, which continues into a canal with a muscular sheath. The worms were identified as ‘turbellarians’ from the family Notenteridae (Fecampiida). This family contains only one species, Notentera ivanovi, reported from the gut of a polychaete at the White Sea. The worms that we found in the gastrointestinal tract of the octopuses were morphologically similar to N. ivanovi but differed from it in several important respects. Phylogenetic analysis based on 28S rDNA gene showed that the newly found worm clustered together with other fecampiids in a highly supported clade and was closely related to N. ivanovi. On the basis of these morphological and molecular data, we described a new species, Octopoxenus antarcticus gen. nov., sp. nov. (Fecampiida: Notenteridae), establishing a new genus to accommodate it and provided an updated diagnosis of the family Notenteridae. This is the first report of a parasitic ‘turbellarian’ from a cephalopod mollusc.

Most gyrodactylids have a haptor armed with a pair of hamuli, two connecting bars and 16 marginal hooks. In some gyrodactylids, however, the haptor is disc-shaped and reinforced by additional sclerites. The genus Polyclithrum has arguably the most elaborate haptor in this group. This study aimed to gain better understanding of the anatomy of Polyclithrum by examining neuromusculature and haptoral armament of Polyclithrum ponticum, a species parasitizing Mugil cephalus in the Black Sea, with emphasis on haptoral sclerites and musculature in connection with host-attachment mechanisms. Musculature was stained by phalloidin, the nervous system by anti-serotonin and anti-FMRFamide antibodies, and haptoral sclerites were visualized in reflected light. The study provided new information on sclerites: in addition to previously described supplementary sclerites (A1–6), ear-shaped sclerites (ESSs) and two paired groups of ribs, reflected light revealed a rod-shaped process on the ESSs and a pair of small posterior sclerites. The sclerites were shown to be operated by 16 muscles, the most prominent of which were two transverse muscles connecting the hamular roots, three muscles attached to sclerite A2, the muscle fibres of anterior ribs and a set of extrinsic muscles. The nervous system consists of a pair of cerebral ganglia connected by a commissure and three pairs of nerve cords that unite in the haptor to form a loop between the opposite cords. The arrangement of sclerites and muscles suggests that Polyclithrum initiates the attachment by clamping a host's surface with longitudinally folded haptor and then secures its position with marginal hooks.

Calodium hepaticum is a zoonotic nematode with a worldwide distribution. Although the host range of C. hepaticum includes a wide spectrum of mammals (including humans), this parasite is predominantly associated with the families Muridae and Cricetidae. Several Sigmodontinae species from Argentina were found to be infected by C. hepaticum, with a high prevalence in Akodon azarae. The present study focuses on C. hepaticum eggs from natural infection of three species of sigmodontine rodents from Argentina. Eggs were genetically characterized (intergenic 18S rRNA region). The objectives of this work are: (i) to propose a new analytical methodology; and (ii) to morphologically characterize C. hepaticum eggs, from three Sigmodontinae species (A. azarae, Calomys callidus and Oligoryzomys flavescens). Analyses were made by the Computer Image Analysis System based on the new standardized measurements and geometric morphometric tools. The resulting factor maps clearly illustrate global size differences in the parasite eggs from the three Sigmodontinae species analysed. The degree of similarity between egg populations was assessed through pairwise Mahalanobis distances, showing that the largest distances were detected between parasite eggs from C. callidus and O. flavescens. Herein, the phenotypical plasticity of C. hepaticum eggs is shown. Significant positive correlations were obtained between each egg parasite principal component 1 and rodent corporal characteristics: weight; liver weight; rodent length; and rodent body condition. The usefulness of the geometric morphometric analysis in studies of the relationship between C. hepaticum and its host must be highlighted. The high prevalence observed in A. azarae, associated with the wide size range of the parasite eggs evidenced by principal component analysis, suggests A. azarae to be the Sigmodontinae host species that plays the most important role as reservoir host for C. hepaticum in the New World.

The helminth infection caused by Strongyloides stercoralis is widespread in tropical regions, but rare in European countries. Unfamiliarity with the disease and diagnostic obstacles could contribute to its lethal outcome. Frequent use of corticosteroids during the COVID-19 pandemic could increase its significance. The aim of this retrospective descriptive study was to explore disease patterns and discuss clinical dilemmas in patients with S. stercoralis hyperinfection treated at the University Hospital for Infectious Diseases ‘Dr. Fran Mihaljević’ in Zagreb, Croatia, between 2010 and 2021. Five out of 22 (22.7%) immunosuppressed patients treated due to strongyloidiasis developed hyperinfection. All patients were male, median 64 years; four were immunosuppressed by corticosteroids (although ileum resection could have been the trigger in one) and one by rituximab. The diagnosis was established after a median of 1.5 months of symptom duration, accidentally in all patients, by visualizing the parasite in the gastric/duodenal mucosa in four cases, and bronchial aspirate in one. All patients were cachectic, four out of five had severe hypoalbuminemia and all suffered secondary bacterial/fungal infection. Despite combined antibiotic, antifungal and antihelmintic therapy, three out of five of the patients died, after failing to clear living parasites from stool samples. We can conclude that significant delays in diagnosis and lack of clinical suspicion were observed among our patients with the most severe clinical presentations of strongyloidiasis. Although being beyond diagnostic recommendations for strongyloidiasis, an early upper gastrointestinal endoscopy with mucosal sample analysis could expedite diagnosis in severe, immunosuppressed patients. The persistence of viable parasites in the stool despite antihelmintic therapy should be further investigated.

In order to identify types of forage that inhibit pasture contamination, an evaluation was performed of the effect of the forage legumes Trifolium repens (white clover), Trifolium pratense (red clover) and Lotus corniculatus (bird's-foot-trefoil) on the survival and migration of infective larvae (L3) of gastrointestinal nematodes (GIN) of sheep. An experimental area of 441 m2 was divided into four blocks, subdivided into areas of 1.20 × 1.20 in which the three forage legumes were separately overseeded. After growth of the forage in each subdivision, experimental units were established that were later artificially contaminated with sheep faeces containing GIN eggs. Between October and December 2018, pasture, faecal and soil samples were collected on four occasions during weeks 1, 2, 4 and 8 after the deposition of faeces. In week 6, the forage legumes in all the experimental units were mown to simulate grazing. The number of L3 was quantified to determine their survival in the pasture, faeces and soil. In addition, the horizontal migration of L3 was measured at two distances from the faecal pellets (10 and 30 cm), as well as their vertical migration at two heights of the plant stems, that is, lower half and upper half. Larvae vertical migration was affected by the forage species (P < 0.001), in that bird's-foot-trefoil contained fewer larvae in the upper stratum. Bird's-foot-trefoil restricted the migration of L3 to the upper stratum of the plant, which could potentially decrease the risk of infection by intestinal nematodes in grazing sheep.

Amphibians are a widespread Chordata taxon and are important for maintaining the balance of both terrestrial and aquatic ecosystems. Brazil has a rich amphibian fauna; however, little is known about the role of their ecology and phylogenetic relationships during the assembly processes of associated endoparasite communities. Herein, we describe an endoparasite community in an anuran assemblage in the Caatinga, a unique biome of dry forests in north-eastern Brazil. We studied endoparasite diversity, as well as the effects of body length, body mass, body volume and sex on parasite abundance. We also investigated the influence of ecological and historical factors and anuran microhabitat use on endoparasite composition. We analysed individuals from 13 anuran species distributed across five families: Odontophrynidae (Proceratophrys cristiceps); Leptodactylidae (Leptodactylus fuscus, Leptodactylus vastus, Leptodactylus macrosternum, Leptodactylus troglodytes and Physalaemus cuvieri); Hylidae (Pithecopus gonzagai, Scinax x-signatus, Boana raniceps and Dendropsophus nanus); Bufonidae (Rhinella diptycha and Rhinella granulosa); and Microhylidae (Dermatonotus muelleri). We found nine species of endoparasites, including seven nematodes (Aplectana membranosa, Cosmocerca sp., Oswaldocruzia mazzai, Raillietnema spectans, Rhabdias fuelleborni, Schrankiana sp. and Physaloptera sp.), one species of Trematoda (Glypthelmins pseudium) and one non-identified cestode. There was no significant relationship between endoparasite abundance and host body length, body mass, body volume and sex. A phylogenetic principal component analysis showed that ecological factors had a greater influence on endoparasite assemblage than historical factors. Similarly, our results showed that ecological factors had a greater influence on anuran microhabitat use compared to historical factors, which contributed to the generalist characteristics presented by most of the sampled endoparasite species.

As part of surveillance of snail-borne trematodiasis in Knowsley Safari (KS), Prescot, United Kingdom, a collection was made in July 2021 of various planorbid (n = 173) and lymnaeid (n = 218) snails. These were taken from 15 purposely selected freshwater habitats. In the laboratory emergent trematode cercariae, often from single snails, were identified by morphology with a sub-set, of those most accessible, later characterized by cytochrome oxidase subunit 1 (cox1) DNA barcoding. Two schistosomatid cercariae were of special note in the context of human cercarial dermatitis (HCD), Bilharziella polonica emergent from Planorbarius corneus and Trichobilharzia spp. emergent from Ampullacaena balthica. The former schistosomatid was last reported in the United Kingdom over 50 years ago. From cox1 analyses, the latter likely consisted of two taxa, Trichobilharzia anseri, a first report in the United Kingdom, and a hitherto unnamed genetic lineage having some affiliation with Trichobilharzia longicauda. The chronobiology of emergent cercariae from P. corneus was assessed, with the vertical swimming rate of B. polonica measured. We provide a brief risk appraisal of HCD for public activities typically undertaken within KS educational and recreational programmes.

The annual migration of birds involves a very large number of inter-continental and intra-continental movements in which thousands of bird species participate. These migrations have been associated with the spread of pathogens worldwide, including bacteria, viruses and parasites. This study describes the case of a black stork (Ciconia nigra) that was ringed at the nest in Latvia and died five months later in the south-east of the Iberian Peninsula. Post-mortem examination revealed that the cause of death was electrocution. In addition, a massive infection by the trematode Chaunocephalus ferox (Digenea: Echinostomatidae) causing severe granulomatous lesions throughout the small intestine was detected. This is the first report of C. ferox infection in a black stork in the Iberian Peninsula, a trematode that, due to the severe lesions it causes, can affect the health of C. ferox-infected wild birds, particularly in severely infected long-distance migrants. The dispersal of platyhelminths associated with migratory birds is discussed. After the ringing at the nest, the black stork was sighted in Central Europe one month before its capture, and the trematodes found by necropsy were mostly mature adults. Consequently, we estimate that this juvenile animal acquired the infection during its migration in a European area other than the Iberian Peninsula, evidencing a long-distance parasite spread through its migratory host. Our study highlights that bird ringing can be used to understand the epidemiological implications that bird migratory behaviour may have on the dispersal of parasites.