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Infection of non-indigenous pumpkinseed (Lepomis gibbosus, Centrarchidae) with the bothriocephalidean cestode Bothriocephalus claviceps (Cestoda: Bothriocephalidae) was confirmed at several sites in the lower Oder river basin in Poland. The preferred host for this cestode species is the European eel (Anguilla anguilla), with a wide range of other fish species serving as paratenic hosts. The pumpkinseed showed a relatively high prevalence and abundance of larvae, along with several mature and gravid specimens, thereby confirming development of B. claviceps to the adult stage in an alternative host. As such, the pumpkinseed may represent an additional definitive host for this cestode, in addition to its role as a paratenic host, as previously recorded in other European regions. Our results indicate that inclusion of pumpkinseed as an additional definitive host in the cestode’s life cycle, high infection of pumpkinseed with B. claviceps larvae and inclusion of pumpkinseed in the eel’s diet could potentially lead to increased parasite pressure on native fish hosts. Further comparative studies or experimental testing will be required to confirm this.

Parasitic nematodes in the genus Dracunculus have a complex life cycle that requires more than one host species in both aquatic and terrestrial habitats. The most well-studied species, Dracunculus medinensis, is the causative agent of human Guinea worm disease (dracunculiasis). There are several other Dracunculus species that infect non-human animals, primarily wildlife (reptiles and mammals). The classic route of D. medinensis transmission to humans is through the ingestion of water containing the intermediate host, a cyclopoid copepod, infected with third-stage larvae (L3s). However, many animal hosts (e.g., terrestrial snakes, dogs) of other Dracunculus sp. appear unlikely to ingest a large number of copepods while drinking. Therefore, alternative routes of infection (e.g., paratenic or transport hosts) may facilitate Dracunculus transmission to these species. To better understand the role of paratenic and transport hosts in Dracunculus transmission to animal definitive hosts, we compared copepod ingestion rates for aquatic species (fish, frogs [tadpoles and adults], and newts) which may serve as paratenic or transport hosts. We hypothesized that fish would consume more copepods than amphibians. Our findings confirm that African clawed frogs (Xenopus laevis) and fish consume copepods, but that fish ingest, on average, significantly higher numbers (68% [34/50]) than adult African clawed frogs (36% [18/50]) during a 24-h time period. Our results suggest that amphibians and fish may play a role in the transmission of Dracunculus to definitive hosts. Still, additional research is required to determine whether, in the wild, fish or frogs are serving as paratenic or transport hosts. If so, they may facilitate Dracunculus transmission. However, if these animals simply act as dead-end hosts or as means of copepod population control, they may decrease Dracunculus transmission., Graphical abstract Image 1, Highlights • Copepod ingestion during 24 h was assessed for fish and amphibians. • Significant numbers of copepods were consumed by fish and adult Xenopus. • Tadpoles and newts did not consume large numbers of copepods during this time. • Fish and amphibians may facilitate Dracunculus transmission. • Further studies may elucidate how copepod ingestion impact parasite transmission.

The global Guinea worm eradication program represents one of the great public health success stories of our time. Since the program’s inception in 1980, the number of annual human cases has reduced by 99.9%, from an estimated 3.5 million cases in 21 countries1 to only 54 cases in three countries at the end of 2019.2,3 Causing significant disability and pain, Guinea worm infection occurs through the ingestion of freshwater copepods (small water crustaceans) infected with stage 3 Dracunculus medinensis larvae. After approximately 10–14 months, a blister forms (usually on a lower limb) where the pregnant adult female Guinea worm ruptures the skin.4 Upon detection of a possible Guinea worm case, patients are transported to health centers where they receive care. The worm is extracted via “controlled immersion,” in which the wound is submerged in a water-filled container to induce worm emergence without the risk of contaminating public water sources.5 Guinea worm is a disease of poverty, primarily impacting people in remote areas who do not have access to safe water for drinking and are forced to drink from stagnant water sources such as ponds, pools, and unprotected open wells that may contain infected copepods.5 The seasonality of disease incidence varies geographically owing to the timing of precipitation patterns and abundance of stagnant water sources.5 For example, in the Sahel (Mali, Niger, Chad), cases historically peaked in the months May through October when stagnant water sources are most abundant. Young adults (15–45 years of age) are most likely to be infected with D. medinensis, although persons of all ages can develop infection.5 Guinea worm cases have historically been equally distributed among men and women, though some exceptions have been noted.5 Occupation is an important risk factor; farmers and persons who fetch drinking water are commonly infected.5 In some countries (i.e., Mali, Niger, and Burkina Faso), certain ethnic groups are at higher risk of infection because of seasonal migration across long distances and, in some cases, the seasonal search for water or pasture for cattle.5 Conventional thought about Guinea worm transmission ecology was challenged in 2012 when canine cases were first reported in Chad in significant numbers,6 spurring new research on transmission dynamics. Today, Chad is the epicenter of D. medinensis transmission, reporting 88% of human cases (N = 48) and virtually all canine cases (99%, N = 1,927) worldwide in 2019.2,3,7 Recent case-control studies conducted in Chad have shown that water sources associated with increased risk for Guinea worm include lagoons, ponds, and untreated water from hand-dug wells.8,9 No associations have been observed to date between human Guinea worm and consumption of fish and frogs,9 which has been demonstrated in dogs.10 This is could be because humans are less likely than dogs to consume uncooked small fish/fingerlings and frogs, which can serve as transport or paratenic hosts.11,12 Over five years have passed since Eberhard et al.6 first described the unusual epidemiological patterns in Chad, noting that human cases are sporadic and rare in comparison with canine cases, with no apparent association with common water sources and no clustering by village. Since that time, the number of canine cases has generally increased steadily as surveillance expanded,2,3,13 whereas human cases have remained generally constant and at low numbers.2,3 Here, we use additional surveillance data to determine whether those epidemiologic patterns persist and to characterize in greater depth human Guinea worm in Chad, with the broader goal of furthering our understanding D. medinensis transmission and epidemiology.

The presence of zoonotic relevant Angiostrongylus cantonensis infections has recently been reported in rat final hosts and gastropod intermediate hosts in Tenerife, Spain. However, data on A. cantonensis, Angiostrongylus vasorum and Aelurostrongylus abstrusus prevalences in endemic gastropods for other islands of the Macaronesian Archipelago are still missing. In order to fill this gap, we conducted an epidemiological study on terrestrial native slug (Plutonia lamarckii) and snail (Cornu aspersum, Theba pisana, Rumina decollata) species in 27 selected locations of Tenerife, Gran Canaria, El Hierro, Lanzarote, La Palma and Fuerteventura. Overall, 131 terrestrial gastropods were collected in winter/spring season 2018/2019 and examined for the presence of metastrongyloid lungworm larvae via artificial digestion. The current data revealed a total prevalence of 4.6% for A. vasorum, 3.8% for A. abstrusus and 0.8% for A. cantonensis. In Tenerife, three lungworm species were detected, thereby re-confirming A. cantonensis endemicity for this island. Prevalences of snails (C. aspersum) originating from El Hierro were 5% for A. abstrusus and 15% for A. vasorum, respectively, with larval burdens up to 290 larvae per specimen. This epidemiological study indicates the presence of human, canine and feline lungworm species in Macaronesia, Spain. The current data—particularly those on anthropozoonotic A. cantonensis—call for a regular large-scale monitoring on intermediate hosts, paratenic hosts and definitive hosts to prevent further spread of lungworm-related diseases in humans and animals.

This study investigated the diversity and ecology of endohelminth parasites in an assemblage of 15 omnivorous and four piscivorous fish species in a tributary of the Amazon River (Brazil). Fish were collected bimonthly to evaluate the diversity and ecology of endohelminth parasites in omnivorous and piscivorous fish species in a tributary of the Amazon River using traditional methods and other methods. A total of 4,234 parasites of 19 species (8 Nematoda, 5 Acanthocephala, 4 Digenea and 2 Cestoda) were sampled from 531 fish specimens. Procamallanus (Spirocamallanus) inopinatus and Contracaecum sp. were the predominant parasite species, and the parasites showed an aggregated dispersion pattern. Positive correlation was shown between the abundance of Contracaecum sp., Hysterothylacium patagonense and P. (S.) inopinatus, and the body size of 15.8% of the host fish examined. No difference was shown for Shannon diversity index and evenness between omnivorous and piscivorous hosts, but the richness, abundance and intensity of parasites were greater in piscivorous host fish. In the fish assemblage, the presence of parasites in the larval and adult stages suggests that these are intermediate, paratenic and definitive hosts, reflecting the feeding habits of the omnivorous and piscivorous host species. 74.1% of the parasite species were new records for the different host species examined. The size of the hosts, trophic level and diet had an influence on the structuring of endohelminth communities and infracommunities.

Toxocara canis, a common roundworm that mainly causes toxocariasis, is a zoonotic parasite found worldwide. Humans, an accidental host, can acquire T. canis infection through accidental ingestion of T. canis-embryonated egg-contaminated food, water, and soil, and by encapsulated larvae in a paratenic host's viscera or meat. Long-term residence of T. canis larvae in a paratenic host's lungs may induce pulmonary inflammation that contributes to lung injury, airway inflammatory hyperresponsiveness, and collagen deposition in mice and clinical patients. This study intended to investigate the relationship between T. canis infection and allergic asthma in BALB/c mice inoculated with high, moderate, and low doses of T. canis eggs for a 13-week investigation. The airway hyperresponsiveness (AHR) to methacholine, collagen deposition, cytokine levels, and pathological changes in lung tissues was assessed in infected mice at weeks 1, 5, and 13 postinfection. The cell composition in bronchoalveolar lavage fluid of infected mice was assessed at weeks 5 and 13 postinfection. Compared with uninfected control mice, all groups of T. canis-infected mice exhibited significant AHR, a dose-dependent increase in eosinophilic infiltration leading to multifocal interstitial and alveolar inflammation with abundant mucus secretion, and collagen deposition in which the lesion size increased with the infective dose. Infected mice groups also showed significant expressions of eotaxin and type 2 T-helper-dominant cytokines such as interleukin (IL)-4, IL-5, and IL-13. Overall, these results suggest that T. canis larval invasion of the lungs may potentially cause pulmonary inflammatory injury and could subsequently contribute to the development of allergic manifestations such as asthma.

Summary The larvae of the genus Baylisascaris can cause larva migrans in mammals and birds. This study investigated the larval migration of Baylisascaris potosis, the roundworm of kinkajou (Potos flavus), in chickens and the associated clinical manifestations of the host. Thirty-six 3-week-old chickens divided into 6 groups were orally inoculated with 3,000 B. potosis eggs/chick. Each group of chicken was necropsied at days 1, 2, 3, 7, 30 and 90 PI (post inoculation), and the number of larvae in various organs were counted until day 90 PI. No clinical signs were observed in chickens during the study. Larvae were detected from the liver, lungs or breast-muscles of 13/36 (36.1%) chickens. The mean total number of larvae in the liver, lungs and breast-muscles at days 1, 2, 3, 7, 30 and 90 PI were 0.34, 0.17, 1.66, 1.01, 0.17 and 0, respectively. No larvae were found in the brain, eyes, hid-limb muscles, heart, kidneys and spleen. Although infectivity of larvae in egg-inoculated chickens was low, the present study demonstrated that B. potosis larvae can migrate in chickens tissues up to day 30 PI. The result suggests that chickens can serve as a paratenic host for B. potosis and may underline a public health importance of B. potosis infection as a potential foodborne disease in humans.

As a result of the experimental infection of rats with metacercariae of Paragonimus heterotremus Chen et Hsia, 1964 from crabs (Potamiscus tannanti) caught in Yen Bai province, Vietnam, it was found that worms migrated into the lungs, to the liver and less frequently to the tissue that lines body cavities of the hosts, where they reached the adult stage, but in the muscles, worms stayed at the larval stage. Studies have shown that for P. heterotremus, rats can simultaneously play the role of the final and paratenic host; herewith, an infection with the trematode of this species can lead to the development of three forms of paragonimiasis: pulmonary, hepatic and muscular. Eggs from the adult worms localised in the liver, unlike eggs from the adult worms localised in the lungs, were not excreted into the external environment, but accumulated inside the organ. Histology and description of changes, which take place on the external surface of organs affected with P. heterotremus, are given in this study. Based on the behavioural characteristics of worms during rat infection and molecular genetic data, we established that worms from Vietnam and India should be assigned to different species of Paragonimus. P. heterotremus distribution is limited to the territory of the Southeast China, Northern Vietnam, Laos and Thailand.

Various species of mammals, including humans and wild boars, can serve as paratenic hosts of Alaria alata mesocercariae – Distomum musculorum suis (DMS). Cases of DMS can be reliably detected by the recent introduction of the A. alata mesocercariae migration technique (AMT). The aim of this study is to present current data on the occurrence of DMS in wild boars in north-eastern Poland, and to compare the findings with those obtained in other European countries. A. alata was identified in 98 of 221 (44.3%) tissue samples of wild boar taken from five provinces in north-eastern Poland during the hunting seasons 2015–2016 and 2016–2017 based on AMT analysis. Positive results were found in all studied regions, but the percentage of infected individuals ranged from 26.3% in the Pomorskie province to 65.5% in the Warmińsko-Mazurskie province. The mean number of larvae exceeded seven larvae per 30 g sample for three provinces: Pomorskie, Mazowieckie and Lubelskie. In turn, lower values were found in the Warmińsko-Mazurskie province (3.3 larvae per 30 g), and the lowest in the Kujawsko-Pomorskie province (1.8 larvae per 30 g). The occurrence and intensity of A. alata infestation in wild boars was found to depend on the environment in which they live. Neither the sex or the age of the wild boar appeared to influence the occurrence nor the intensity of infestation., Graphical abstract Image 1, Highlights • High percentage (44.3) of Alaria alata in samples of wild boars in Poland. • High prevalence of A. alata infestation in wild boars compared to other reports. • Sex does not affect the risk or intensity of infestation by A. alata in wild boars. • Age does not affect the risk or intensity of A. alata infestation in wild boars. • A. alata poses a potential risk for wild boar consumers in Poland.

Domestic dogs can function as either paratenic or definitive hosts for the zoonotic raccoon roundworm Baylisascaris procyonis. However, factors leading to development of patent infections in dogs are under-studied. Here we compared infection dynamics of B. procyonis in dogs vs the natural raccoon host. Dogs and raccoons were inoculated 5000 or 500 B. procyonis eggs (n = 3 per dose) or were fed B. procyonis-infected laboratory mice (n = 3 per dose; mice inoculated with 1000 or 250 eggs). Fecal samples were analysed via flotation and a commercial coproantigen ELISA designed for detection of Toxocara spp. Two of 12 dogs (both received low dose larvae) developed patent infections; all 12 raccoons became infected with 10 developing patent infections. Compared with dogs, prepatent periods were shorter in raccoons and maximum egg outputs were much greater. Baylisascaris procyonis coproantigens were detectable via ELISA in all raccoons and the patently infected dogs. Finally, dogs spontaneously lost infections while all patently infected raccoons shed eggs until conclusion of the study. Our results demonstrate that dogs are clearly suboptimal hosts showing limited parasite establishment and fecundity vs raccoons. Despite the low competence, patently infected dogs still pose a risk for human exposure, emphasizing the importance of control measures.

International audience; Bivalve molluscs are now considered indicator species of aquatic contamination by human parasitic protozoa. Nonetheless, the possible effects of these protozoa on the immune system of their paratenic hosts are poorly documented. The aim of this study was to evaluate the effects of two protozoa on hemocyte viability and phagocytosis from two mussels, the zebra mussel (freshwater habitat) and the blue mussel (seawater habitat). For these purposes, viability and phagocytic markers have been analysed on hemocytes from mussels without biological stress (control hemocytes), and on hemocytes exposed to a biological stress (Toxoplasma gondii and Cryptosporidium parvum oocysts). We report, for the first known time, the interactions between protozoa and hemocytes of mussels from different aquatic environments. Zebra mussel hemocytes showed a decrease in phagocytosis of fluorescent microbeads after exposure to both protozoa, while blue mussel hemocytes reacted only to T. gondii oocysts. These decreases in the ingestion of microbeads can be caused by competition between beads and oocysts and can be influenced by the size of the oocysts. New characterisations of their immune capacities, including aggregation, remain to be developed to understand the specificities of both mussels.

Clarias gariepinus is a fish from North of South Africa and was later introduced in several countries, including Brazil. The present study aimed to describe the first report of Diplostomidae metacercariae in C. gariepinus in Brazil. For this, 30 C. gariepinus were captured in a lake in the city of Campos dos Goytacazes, RJ, Brazil. Fishes were euthanized using freezing and necropsied for collection of parasites. The organs were dissected and analyzed for the presence of parasites that were processed for light and scanning electron microscopy. Trematodes were stained with Semichon’s Carmine and Gomori’s Trichrome, observed and schematized under a light microscope with image analysis software. A total of 190 trematodes were collected from the gills, suprabranchial organs, heart, stomach, intestinal mesentery, liver and body cavity of the fish. The parasites had a foliaceous body divided by a discrete constriction, without genital primordia, and a holdfast organ present at the posterior region, typical of metacercariae of the family Diplostomidae. It was classified as the 'Diplostomulum' morphotype, based on the morphology of the reserve bladder structure. This is the first report of the metacercariae of Diplostomidae parasitizing C. gariepinus in the Americas. This fish acts as an intermediate or paratenic host of this digenean in Brazil. Resumo Clarias gariepinus é um peixe do Norte da África do Sul e posteriormente introduzido em diversos países, incluindo o Brasil. O presente estudo teve como objetivo descrever o primeiro relato de metacercárias em C. gariepinus no Brasil. Para tanto, 30 C. gariepinus foram capturados em um lago na cidade de Campos dos Goytacazes, RJ, Brasil. Os peixes foram eutanasiados por congelamento e necropsiados para a coleta de parasitos. Os órgãos foram dissecados e analisados para a presença de parasitos que foram processados para microscopia ótica e electronica de varredura. Os Trematodeos foram corados com carmim de Semichon e tricrômico de Gomori, observados e esquematizados sob microscópio ótico equipado com software de análise de imagens. Um total de 190 trematodeos foram coletados das brânquias, órgãos suprabrânquiais, coração, estômago, mesentério intestinal, fígado e cavidade corporal dos peixes. Os parasitos tinham corpo foliáceo, dividido por uma discrete contrição, sem primórdio genital, e órgão tribocítico presente na região posterior, típico das metacercárias da fámilia Diplostomidae. Foram classificados como do morfotipo 'Diplostomulum', baseado na morfologia da estrutura da bexiga de reserva. Este é o primeiro relato de metacercárias de Diplostomidae parasitando C. gariepinus nas Américas. Este peixe atua como hospedeiro intermediário ou paratênico deste Digenea no Brasil.

Little information is available on host-parasite relationships between bivalves and larval nematodes. Herein, we describe nematode larvae (likely stage 2) in the infraorder Ascaridomorpha infecting the foot, intestine, and mantle of a freshwater mussel (Alabama rainbow, Villosa nebulosa [Conrad, 1834]) and detail histopathological changes to infected tissues. A total of 43 live mussels from the South Fork of Terrapin Creek, Alabama, were collected between 2010 and 2014, with 14 sectioned for histopathology and 29 dissected. Of the 14 sectioned mussels, 5 appeared to be uninfected, and 7, 1, and 1 had histozoic infections observed in the foot and intestine, intestine only, and mantle edge and foot, respectively. Twenty-three of 29 (79%) of the mussels dissected were infected by live nematodes, and mean nematode abundance was 8.3 (CL = 5.23–13), with 2 mussels infected with >100 nematodes each. Thus, with a total of 32 of the 43 collected mussels observed with nematodes, overall infection prevalence was 74.4% (CL = 0.594–0.855). The 18S rDNA of this nematode was 99% similar to that of several ascaridids (species of Kathlaniidae Lane, 1914 and Quimperiidae Baylis, 1930) that mature in aquatic/semi-aquatic vertebrates; the recovered 18S phylogenetic tree indicated this nematode from V. nebulosa shares a recent common ancestor with Ichthyobronema hamulatum (Ascaridomorpha: Quimperiidae; GenBank Accession Number KY476351). Pathological changes to tissue associated with these infections comprised focal tissue damage, but a cellular response was not evident. The Alabama rainbow possibly represents an intermediate or paratenic host. Given these results, the nematode is likely not pathogenic under normal stream conditions; however, high intensity infections in the foot could inhibit pedal extension and retraction; which would have demonstrable health consequences to a freshwater mussel. Based on our review of the bivalve mollusc parasite literature, a collective biodiversity of 61 nematodes reportedly exhibit some degree of symbiosis (from commensal to parasitic) with 21 bivalves (28 nematode spp. from 17 marine bivalve spp.; 33 nematode spp. from 4 freshwater bivalve spp.); only four records exist of putatively parasitic nematodes from Unionida. The present study represents the first description of a nematode species that invades the tissues of a Unionidae species., Graphical abstract Image 1, Highlights • Pathological changes comprised focal tissue damage but a cellular response to the parasites was not evident. • Nematodes from infected tissues appear to be second stage larvae of Ascaridomorpha. • The small subunit rDNA sequence of this nematode was 99% similar to species of Kathlaniidae and Quimperiidae. • The nematode is likely not pathogenic under normal stream conditions.

Purpose of Review: In this review, we summarized information regarding a neglected parasitic infection by zoonotic helminths that cause the cutaneous larva migrans, especially A. braziliense, A. caninum and Uncinaria stenocephala, among other associated or potentially linked to cutaneous larva migrans (CLM), a paratenic infection. Recent Findings: Although there has been an increase in the number of publications on CLM over the last decades, this is still a neglected condition among tropical diseases, with not more than 350 papers per year combined in databases such as PubMed. Although the disease is considered a limited dermatosis, systemic invasion with Löeffler’s syndrome have been exceptionally described. Summary: CLM affects mainly the feet, but any part of the body in contact with contaminated soil or sand can be involved, becoming especially a common problem for tourists on beach holidays where they walk on bare feet or lie on infested sand.

The scarce information on the helminth fauna in otariids from the Southeastern Pacific comes mainly from stranded individuals or killed for that purpose. In this study, we compared the abundance and composition of enteroparasitic assemblages of Otaria flavescens using coprological techniques. Three sampling localities from north to south spanning 2,200 km off the Chilean coast were considered (Iquique, Vin a del Mar, and Talcahuano). In all, 60 fecal samples were collected, and eggs belonging to 5 taxa were found in 91.6% of the samples. They were the anisakid nematodes Contracaecum and Pseudoterranova, the cestode Adenocephalus (syn. Diphyllobothrium), the trematode Ogmogaster, and the acanthocephalan Corynosoma. Samples from southern Chile (Talcahuano) showed the highest prevalence. Adenocephalus eggs had the highest prevalence and abundance in Iquique and Talcahuano, whereas Ogmogaster was the less prevalent and abundant in all sampling localities. Corynosoma eggs had similar prevalence and abundance among sampling localities, and Pseudoterranova eggs were absent in Iquique and with median prevalence values in Vin a del Mar and Talcahuano. Thus, the composition of parasite egg assemblages was different between sampling localities. These differences between sampling localities may help to explain differential records of some zoonotic parasitoses such as pseudoterranovosis and diphyllobothriosis in Peru and Chile, where consumption of raw or marinated fish (ceviche) is common. For example, the lower diversity of parasite egg assemblages in the northern Chilean coast may be due to the absence or lower abundance of first intermediate/paratenic hosts of Pseudoterranova.

International audience; Based on light and electron microscopical studies, a new nematode parasite, Echinocephalus inserratus sp. n. (Spirurida: Gnathostomatidae), is described from the spiral valve of the broad cowtail stingray Pastinachus ater (Macleay) (Dasyatidae, Myliobatiformes) from off New Caledonia. The new species is morphologically and biometrically most similar to Echinocephalus overstreeti Deardorff et Ko, 1983, differing from it mainly in the absence of serrations on the posterior parts of pseudolabia and on interlabia, and in having a longer gubernaculum (150-299 µm long). Morphologically unidentifiable, mostly encapsulated larvae of Echinocephalus spp. were recorded from the following six species of teleost fishes collected in New Caledonian waters, serving as paratenic hosts: Perciformes: Acanthopagrus berda (Forsskål) (Sparidae) and Nemipterus furcosus (Valenciennes) (Nemipteridae); Tetraodontiformes: Abalistes stellatus (Anonymous), Pseudobalistes fuscus (Bloch et Schneider) (both Balistidae), Lagocephalus sceleratus (Gmelin) (Tetraodontidae) and Aluterus monoceros (Linnaeus) (Monacanthidae). Co-parasitising larvae of Ascarophis sp. and Hysterothylacium sp. were also collected from P. fuscus. All these findings represent new host and geographical records. A key to valid species of Echinocephalus Molin, 1858 is provided.

Rhadinorhynchus trachuri Harada, 1935 is probably one of the most widely distributed acanthocephalans in the world, but the least known taxonomically. This report fills the large gap of knowledge about its taxonomy, host relationships, and worldwide distribution in marine fishes of both sides of the Pacific Ocean and the connecting Indian Ocean. To revisit the taxonomy of this species, provide a more complete morphological description and expand our knowledge about its morphological identity and variations and elaborate on its worldwide zoogeography; accounting for its worldwide distribution based on new collections, museum specimens, and on published reports. Specimens from Vietnam were processed, identified, and documented using standard techniques. Specimens of R. trachuri were studied from three species of fish in three families, Auxis thazard (Lacepede) (Scombridae), Megalaspis cordyla (Linn.) (Carangidae), and Tylosurus sp. (Belonidae) in Nha Trang and Binh Thuan in the south Pacific coast of Vietnam and additional museum specimens mostly from salmonids from the Pacific coast of California were also studied. Extensive research was conducted on published records of R. trachuri and its marine fish hosts worldwide. Specimens of R. trachuri are characterized by cylindrical trunk with anterior rings of spines and posterior ventral and lateral spines, a proboscis usually with 12 hook rows each with 19–24 hooks, variable position of testes, and a long uterus, uterine bell attached to the dorsal trunk wall, rounded vagina, and unusually dorsal gonopore. The hosts and geographical distribution of R. trachuri suggest a northern linkage of infections between the Asian and American Pacific populations across the Bering Sea involving salmonid fishes and a southern linkage through the Indian Ocean involving Carangid fishes. Other hosts involved in the cycling and distribution of infections are also discussed with some definitive hosts also serving as paratenic hosts. This is the first complete description about R. trachuri and the first comprehensive accounting of its worldwide distribution in 19 species of marine fish in seven families. Considerable intraspecific variability related to geographical restrictions, intermediate and definitive host specificity and distribution, and host feeding behavior was documented.

The main fish host reaction to an infection with third stage anisakid nematode larvae is a response in which host immune cells (macrophages, granulocytes, lymphocytes) in affected internal organs initially are attracted to the parasite whereafter fibroblasts may enclose the parasite forming granuloma. Generally, the reaction is non-lethal to the parasite which may survive for years in the fish host retaining infectivity to the final host. This may also apply for the anisakid nematode Contracaecum rudolphii (having the adult stage in cormorants, using copepods as first intermediate/paratenic host and zooplankton feeding fish as paratenic hosts). The present study has shown that most Contracaecum rudolphii larvae survive in bream (Abramis brama) (from Lake Balaton, Hungary) whereas the majority of the nematode larvae die in Cyprinus carpio (from Lake Hévíz, directly connected to Lake Balaton). Both cyprinid host species interacted with the nematode larvae through establishing a marked cellular encapsulation around them but with different effects. The differential survival in common carp and bream may theoretically be explained by ecological factors, such as the environmental temperature which either directly or indirectly affect the development of nematode larvae, and/or intrinsic host factors, such as differential immune responses and host genetics.

The common dog roundworm Toxocara canis can infect other animals and humans which may act as their paratenic and accidental hosts, respectively. Larvae do not further develop to adult worms in these hosts. Instead, they undergo migration to various body organs, causing the neglected parasitic disease known as toxocariasis. Although rats are considered as potential paratenic hosts of the parasite, there are only few studies which utilized Rattus norvegicus (Sprague–Dawley strain) for experimental infections involving toxocariasis. This study aimed to determine whether T. canis could establish in Sprague–Dawley rats artificially infected with 500 T. canis embryonated eggs and if the animals can be used as animal models for toxocariasis. Following squash method and tissue digestion, larvae were recovered from the lungs, liver and brain of the infected rats. Furthermore, gross examination of organs revealed macroscopic lesions and hemorrhages in the lungs and brain. Microscopically, accumulation of inflammatory cells, thickening of alveoli lining and destruction of bronchial walls and hepatic necrosis were observed. This study showed that T. canis has established in Sprague–Dawley rats and could serve as a model for Toxocara infection studies.

Cod (Gadus morhua), an important fish species in the Baltic Sea, is the paratenic host for many parasite species, including the zoonotic nematodes, Anisakis sp. and Contracaecum osculatum. We aimed to identify which invertebrate species (found in situ in the fish stomach) are responsible for infection of cod with zoonotic nematodes. We found that Crangon crangon and Gammarus sp., both invertebrate prey species of cod, were infected with Anisakis simplex and C. osculatum, respectively. These host–parasite systems are reported here for the first time, implicating C. crangon and Gammarus sp. as sources of infection of Baltic cod with zoonotic nematodes.

Cystoisospora belliis a coccidian parasite of humans, with a direct fecal–oral transmission cycle. It is globally distributed, but mainly found in tropical and subtropical areas. Many cases ofC. belliinfections have been reported in patients with HIV, and in patients undergoing immunosuppressive therapy for organ transplants or those treated for tumours worldwide. Unsporulated or partially sporulated oocysts ofC. belliare excreted in feces. When sporulated oocysts in contaminated water or food are ingested, asexual and sexual stages ofC. belliare confined to the epithelium of intestines, bile ducts and gallbladder. Monozoic tissue cysts are present in extra-intestinal organs (lamina propria of the small and large intestine, lymph nodes, spleen, and liver) of immunosuppressed humans. However, a paratenic host has not been demonstrated.Cystoisospora belliinfections can be persistent, lasting for months, and relapses are common; the mechanism of relapse is unknown. Recently, the endogenous stages ofC. belliwere re-examined and attention was drawn to cases of misidentification of non-protozoal structures in the gallbladder of patients asC. belli. Here, we review all aspects of the biology ofC. belli, including morphology, endogenous stages, prevalence, epidemiology, symptoms, diagnosis and control.

We analyzed an 11-year time series (2005–2015) of parasite abundance for three intestinal nematode species in the red fox (Vulpes vulpes) as a function of the multi-annual rodent population cycle in low-arctic Norway, while correcting for other potential covariates that could influence prevalence and abundance. Rodents are paratenic and facultative intermediate hosts for the two Ascarididae species Toxascaris leonina and Toxocara canis, respectively and key prey for the red fox. Still the relative importance of indirect transmission through rodents and direct transmission through free-living stages is unclear. Abundance of these Ascarididae species in individual red foxes (N = 612) exhibited strongly cyclic dynamics that closely mirrored the 4-year rodent cycle. Negative binomial models provided evidence for a direct proportional increase in Ascarididae abundance with rodent density suggesting that predator functional response to rodent prey is the key transmission mechanism. In contrast, no cycles and constantly very low abundance were apparent for Uncinaria stenocephala – a third nematode species recorded without paratenic or intermediate stages., Graphical abstract Image 1, Highlights • Long-term study identified host-parasite relationship in low-arctic red foxes. • Indirect transmission through rodent an important pathway for Ascarididae spp. • Multi-annual rodent population cycle key determinant of Ascarididae spp. prevalence.

Anisakidosis is a zoonotic parasitosis induced by members of the family Anisakidae. The anisakid genera includes Anisakis, Pseudoterranova, Hysterothylacium and Contracaecum. The final definitive hosts of these nematodes are marine mammals with a complex life cycle. These nematode parasites use different crustaceans and fish species as intermediate or paratenic hosts and humans are accidental hosts. Human anisakiasis, the infections caused by members of the genus Anisakis, occurs, when seafoods, particularly fish, contaminated with the infective stage (third stage larvae [L3]) of this parasite, are consumed. Pseudoterranovosis, on the other hand is induced by members of the genus Pseudoterranova. These two genera of anisakids have been implicated in human disease globally. There is a rise in reports of gastro-intestinal infections accompanied by allergic reactions caused by Anisakis simplex and Anisakis pegreffii. This review provides an update on current knowledge on Anisakis as a food-borne parasite with special focus on the increasingly reported diversity of fish and crustacean hosts, allergens and immunological cross-reactivity with invertebrate proteins rendering this parasite a significant public health issue., Graphical abstract Image 1, Highlights • Anisakis, is a foodborne zoonotic parasite. • Humans are accidental hosts of Anisakis L3 larva. • Consumption of Anisakis parasite-contaminated seafood causes human anisakiasis. • Zooplankton (Crustaceans: Krills, Squids, Crayfish) composition in fishing regions contribute to Anisakis parasitosis. • Anisakis is the parasite with the largest number of registered allergens with the IUIS.

The acanthocephalan parasite Plagiorhynchus cylindraceus has a global distribution and utilizes isopods and birds as intermediate and definitive hosts, respectively. Occasionally, mammals of various orders can act as paratenic hosts. In hedgehogs, severe cases have been reported in juvenile specimens due to secondary infections, as a consequence of complete penetrations of the intestinal wall by cystacanths. In a 66-month study period, we found seven larvae of this parasite encysted in both, the peritoneal cavity and intestine of the Algerian hedgehog, Atelerix algirus in Majorca. Morphology alone was insufficient to identify the species, due to the lack of previous reports and taxonomy-informative characters. In the present report, we combined the use of morphology and the DNA-barcoding approach to confirm to identify cystacanths as P. cylindraceus. This is the first report of this parasite in this hedgehog species. The epidemiological implications will be discussed, including the risk of zoonosis and the importance of using modern approaches to identify immature acanthocephalan larvae in wildlife hosts.

Nematodes are an extremely diverse and speciose group of parasites. Adult dracunculoid nematodes (Superfamily Dracunculoidea) occur in the tissues and serous cavities of mammals, fish, reptiles, amphibians and birds. Of the dracunculid group, perhaps best known is Dracunculus medinensis, the human Guinea Worm. Considerable work has been done on D. medinensis; however recent infections in peri-domestic dogs and the finding of naturally-infected paratenic hosts (previously unreported for D. medinensis) indicate we still have much to learn about these parasites. Furthermore, among eight species in the Old World and six species in the New World there is a lack of general life history knowledge as well as questions on species occurrence, host diversity, and transmission dynamics. Herein, we provide a comprehensive review of the genus Dracunculus, in order of a theoretical evolutionary progression from reptilian to mammalian hosts. Species descriptions, where available, are provided but also show where gaps occur in our knowledge of various species. Additionally, many first reports of Dracunculus spp. were done prior to the development and use of molecular tools. This is especially important for this group of parasites as speciation based on morphology is only applicable to males of the genus, and males, given their size, are notoriously difficult to recover from definitive hosts. Therefore, we also discuss current molecular tools used in the investigation of this group of parasites. Given recent host-switching events, the dracunculids are of increasing importance and require further work to expand our understanding of this genus. Keywords: Dracunculus spp, Wildlife, Definitive host, Paratenic host, Intermediate host

The life cycle of Dioctophyma renale involves an intermediate host (oligochaete), a paratenic hosts (fish and frogs), and a definitive host (mustelids and canids). Dogs are at risk of infection with D. renale when they consume paratenic hosts infected with the larval form of D. renale. Water containing the oligochaete intermediate host cannot be disregarded as another source of infection. Infections occur mainly in the right kidney, but worms have also been found in the abdominal cavity as well as other organs. Most dogs appear asymptomatic and infections are usually noted as incidental findings on necropsy. Recently, the Ontario Society for the Prevention of Cruelty to Animals (SPCA) and Humane Society conducted transports of dogs located in northern remote communities. In 2016, some female dogs were found to be infected with D. renale upon ovariohysterectomy. In response to this discovery, we developed a screening protocol to screen for D. renale infections. In 2018, a total of 130 intact dogs were transferred from 2 northern communities in the provinces of Ontario and Manitoba. A prevalence of 7.94% (95% confidence interval 3.87-14.11%) was found from dogs from the northern communities. The screening protocol we developed provides a method of screening for dogs that are transported from communities that could be at risk of infection with D. renale.

Background Toxocariasis, caused by a nematode species of the genus Toxocara, has been described as one of the most prevalent zoonotic helminthiases worldwide. Human transmission may occur by ingesting Toxocara spp. larvae from raw or undercooked meat or organs; however, no comprehensive serosurvey study has been conducted to date investigating the role of cattle as paratenic hosts. The aim of the study reported here was to assess the prevalence of anti-Toxocara spp. antibodies and associated risk factors in bovines from two slaughterhouses located in Presidente Prudente, southeastern Brazil. Methods Blood samples were collected and tested by indirect enzyme-linked immunosorbent assay (ELISA). Cattle farmers voluntarily responded to an epidemiologic questionnaire. Results Overall, 213 of the 553 (38.5%) bovine samples were assessed as seropositive for anti-Toxocara spp. antibodies by indirect ELISA. Multivariate analysis revealed that the source of beef cattle and the presence of dogs or cats at the farm were associated with seropositivity. The use of feedlot systems was associated with lower likelihood of seropositivity. Conclusions These results indicate a high level of anti-Toxocara seropositivity in slaughterhouse cattle, with potentially contaminated meat posing an infection risk to humans. In addition, the presence of dogs and cats where the slaughtered beef cattle were raised was statistically associated with bovine seropositivity, probably due to the overlapping environment at the farm and the lack of pet deworming. The use of feedlot systems was a protective factor likely due to the absence of dog and cat contact, elevated feeding troughs that avoid contact with contaminated soil or grass, and younger age at slaughter of feedlot cattle. In summary, bovines may be used as environmental sentinels of Toxocara spp. contamination, and high seropositivity of slaughterhouse cattle may indicate a potential risk of human toxocariasis through the ingestion of raw or undercooked contaminated meat. Graphical Abstract

Guinea worm (Dracunculus medinensis) has exerted a high human health burden in parts of Africa. Complete eradication of Guinea worm disease (dracunculiasis) may be delayed by the circulation of the parasite in domestic dogs. As with humans, dogs acquire the parasite by directly ingesting infected copepods, and recent evidence suggests that consuming frogs that ingested infected copepods as tadpoles may be a viable transmission route (paratenic route). To understand the relative contributions of direct and paratenic transmission routes, we developed a mathematical model that describes transmission of Guinea worm between dogs, copepods and frogs. We explored how the parasite basic reproductive number (R0) depends on parameters amenable to actionable interventions under three scenarios: frogs/tadpoles do not consume copepods; tadpoles consume copepods but frogs do not contribute to transmission; and frogs are paratenic hosts. We found a non-monotonic relationship between the number of dogs and R0. Generally, frogs can contribute to disease control by removing infected copepods from the waterbody even when paratenic transmission can occur. However, paratenic transmission could play an important role in maintaining the parasite when direct transmission is reduced by interventions focused on reducing copepod ingestion by dogs. Together, these suggest that the most effective intervention strategies may be those which focus on the reduction of copepods, as this reduces outbreak potential irrespective of the importance of the paratenic route.

Paragonimiasis is a particular foodborne parasitic disease that is endemic to southern Kyushu, including Kagoshima Prefecture, Japan. We previously detected Paragonimus westermani triploid larvae in meat samples obtained from wild boars and sika deer hunted in Akune City, Kagoshima Prefecture. These mammals act as paratenic hosts and their meat is a source of human paragonimiasis. Paratenic host mammals and humans become infected with the lung fluke, P. westermani, following consumption of second intermediate hosts, freshwater crab species, namely, Geothelphusa dehaani or Sawagani in Japanese, and Eriocheir japonica or Mokuzugani in Japanese. Therefore, this study aimed to investigate the current infection status of P. westermani in freshwater crabs in Akune City. We collected freshwater crabs from 15 locations and found that the prevalence of P. westermani metacercariae was 1.6% for Sawagani (15 of 941 examined) and 22.1% for Mokuzugani (21 of 95 examined). Based on the morphological characterization of metacercariae and molecular analyses of the internal transcribed spacer 2 region and mitochondrial 16S rRNA gene region using PCR-restriction fragment length polymorphism and sequencing, all metacercariae were identified as the triploid form of P. westermani. These results indicate that Sawagani and Mokuzugani serve as second intermediate hosts to maintain the life cycle of triploid P. westermani. Further, infection in crabs potentially leads to subsequent P. westermani infections in wild mammals, including wild boars and sika deer, both of which are considered important types of game meat in Japan.

Background Spirometra erinaceieuropaei is a diphylobothriid tapeworm with a complex life-cycle including definitive, intermediate and paratenic (transport) hosts. Multiple routes of parasite transmission often make it impossible to determine what type of host a specific infected animal is considered to be. Spargana larvae cause sparganosis, a severe food- and water-borne disease mainly found in Asia. In Poland, Spirometra sp. was reported in large carnivores in Białowieża Primeval Forest for the first time in the 1940s and was recently confirmed as S. erinaceieuropaei in several mammals and snakes using molecular methods. Methods In total, 583 carcasses of 9 carnivore species were necropsied between 2013 and 2019 in north-eastern (NE) Poland. The larvae of S. erinaceieuropaei (spargana) were isolated from subcutaneous tissue, counted, and preserved for genetic analyses. We calculated the prevalence and intensity of infection. To assess spatial variation in S. erinaceieuropaei infection probability in NE Poland, we applied a generalized additive model (GAM) with binomial error distribution. To confirm the species affiliation of isolated larvae, we amplified a partial fragment of the 18S rRNA gene (240 bp in length). Results Spirometra larvae were found in the subcutaneous tissue of 172 animals of 7 species and confirmed genetically as S. erinaceieuropaei. The overall prevalence in all studied hosts was 29.5% with a mean infection intensity of 14.1 ± 33.8 larvae per individual. Native European badgers and invasive raccoon dogs were characterized by the highest prevalence. An analysis of parasite spread showed a spatially diversified probability of infection with the highest values occurring in the biodiversity hot spot, Białowieża Primeval Forest. Conclusions Our study revealed that various mammal species (both native and non-native) can serve as S. erinaceieuropaei reservoirs. The frequency and level of infection may differ between selected hosts and likely depend on host diversity and habitat structure in a given area. Further studies are needed to assess the distribution of the parasite throughout Europe and the environmental and biological factors influencing infection severity in wild mammals.

We found juveniles of Paragonimus in the urinary bladder of a Japanese toad (Bufo japonicus formosus) captured in Kyoto. These were molecularly identified as Paragonimus skrjabini miyazakii Kamo, Nishida, Hatsushika et Tomimura 1961. This is the first report of P. s. miyazakii found in anuran hosts in Japan, indicating that anurans can be paratenic hosts of P. s. miyazakii, as is also the case for Paragonimus skrjabini skrjabini in China. This finding suggests that definitive hosts of P. s. miyazakii can be infected by eating not only crabs or mammal paratenic hosts, but also anurans.

The prevalence and diversity of parasitic nematodes in wildlife have been well studied for certain species, yet for others considerable gaps in knowledge exist. The parasitic nematode Dracunculus insignis infects North American wildlife, and past research on this species has led to an increased understanding of the potential host diversity and transmission of the closely related human Guinea worm, Dracunculus medinensis (which is currently the focus of a global eradication program). Many definitive hosts have been documented for D. insignis; however, the life cycle has been studied only in laboratories, and only a single phylogenetic study has been conducted on D. insignis (from Canada). The goals of the present study were to investigate the prevalence of infections with Dracunculus species among wildlife at a single site (Di-Lane plantation) in the southeastern United States, evaluate the genetic diversity of parasites at this site, and investigate potential paratenic hosts that may be involved in transmission. Over 3 yr, we sampled 228 meso-mammals, reporting an overall prevalence of infection with Dracunculus insignis of 20% (46/228). Amphibians and fish were sampled in the same geographic area as infected meso-mammals. Dracunculus insignis third-stage larvae were recovered from 2 different species of amphibians, but all fish sampled were negative. Phylogenetic analysis of the partial cytochrome c oxidase I (COI) gene showed very little diversity of Dracunculus at Di-Lane; however, we did recover a single nematode from a Virginia opossum (Didelphis virginiana) that falls outside of the D. insignis clade, more closely aligns with Dracunculus lutrae, and may represent an undescribed species. This work documents the occurrence of D. insignis in frogs, a potential transmission pathway for D. insignis at a single geographic site in nature. When applied to the global Guinea Worm Eradication Program, and Chad, Africa, in particular, this work increases our knowledge of the potential role of aquatic animals in the transmission of Dracunculus species and informs on potential intervention strategies that may be applied to the eradication of Guinea worm in Africa.

Based on sequence homology and phylogenetic tree results, the first report of eyeworm Oxyspirura species larvae has been confirmed in a human patient from Vietnam. However, important information related to Oxyspirura larvae was not presented in the case study. This comment provides a more detailed comparison of the Oxyspirura larvae found in the human case study to the avian eyeworm Oxyspirura petrowi.

Graphical abstract, Highlights • Transmammary transmission of Troglostrongylus brevior in domestic cats is suggested. Snails, rats and lizards play a role in the epidemiology of this feline lungworm. Observational parasitology is important in studying events that may occur in small ecological niches around us., Feline lungworms such as Aerulostrongylus abstrusus and Troglostrongylus brevior are snail-borne pathogens causing respiratory disease in domestic cats. Paratenic hosts such as rodents and reptiles have also been implicated in the epidemiology of these parasites. Although A. abstrusus has been recognized for a long time as the most prevalent lungworm among cats worldwide, T. brevior is of major concern in kittens. Bearing in mind that disease due to T. brevior occurs mainly in pediatric patients younger than 6 months of age, the diagnosis of this parasite in two kittens presenting severe respiratory disease from the garden of one of the authors inspired us to investigate the potential routes of transmission for T. brevior in domestic cats. Of the three queens (A, B and C) that delivered kittens (n = 8), only cat A was positive for T. brevior, presenting her two kittens severe respiratory clinical signs, which lead to the exitus in one of them, 18 days of age. In addition, three kittens, the offspring of queen B, turned to be positive at the coprological examination after suckling from queen A, whereas those from queen C (that suckled only on their own mother) remained negative. A series of coprological, histological and molecular tests were conducted to confirm the presence of T. brevior in the patients as well as in the other cats cohabiting the same garden. Adult nematodes were retrieved from the trachea and bronchi of the dead kitten (kitten 1A), and larvae at the histology of the lung and liver parenchyma associated with bronco pneumonitis and lymphocytic pericholangitis, respectively. Cornu aspersum (n = 60), Eobania vermiculata (n = 30) snails (intermediate hosts) as well as lizards and rats (potential paratenic hosts) were collected from the same garden and processed through tissue digestion and molecular detection. Troglostrongylus brevior larvae were recovered through tissue digestion from two C. aspersum (3.33%) and it was confirmed by PCR-sequencing approach, which also detected T. brevior DNA in the liver and lungs of one rat and one gecko lizard. During the COVID-19 lockdown, when scientists spent more time at home, we grasp the opportunity to decipher T. brevior biology and ecology starting in a small ecological niche, such as the garden of our house. Data herein presented led us to suggest: i) the transmammary transmission of T. brevior in domestic cats; ii) the role of intermediate and paratenic hosts (including reptiles) in the epidemiology of the infection which they transmit; as well as iii) the importance of observational parasitology in studying any event that certainly occurs in small ecological niches, as it could be in our home gardens.

Diphyllobothriid tapeworms of the genus Spirometra are causative agents of sparganosis, food-borne zoonotic parasitic disease. They have been recorded in broad spectrum of hosts, including humans, in all continents except Antarctica. Spirometra tapeworms have been intensively studied in several Asian countries; however, they have been rather neglected in Europe. The aim of this study was to provide a pilot screening of Spirometra spp. in Latvia, where data on sparganosis are not available. Tapeworms morphologically identified as diphyllobothriid species were isolated from grey wolves Canis lupus and Eurasian lynxes Lynx lynx from Latvia during the hunting periods 2013–2019. The parasites were subjected to molecular genotyping using sequences of the partial large (LSU rDNA; 615 bp) and small (SSU rDNA; 720 bp) subunits of the nuclear ribosomal RNA gene and complete (1566 bp) cytochrome c oxidase subunit I gene of the mitochondrial DNA (cox1 mtDNA). Analyses of both ribosomal subunits of 13 tapeworms revealed no intraspecific variation within the respective rDNA subunits. On the other hand, sequence analysis of mitochondrial cox1 revealed intraspecific polymorphism displayed by 12 cox1 haplotypes. Comparison of the current data with sequences of the corresponding DNA regions deposited in the GenBank revealed 99.3–99.5% (LSU rDNA), 99.2% (SSU rDNA) and 99.6–100% (cox1 mtDNA) identity of studied tapeworms with Spirometra erinaceieuropaei, which provided the first confirmation of this diphyllobothriid tapeworm in Latvia. Since S. erinaceieuropaei is probably prevalent in Latvian wildlife and may also occur in other potential host species, further studies are needed in order to acquire complex data on its geographic distribution and transmission in the natural environment of Latvia, as well as on the spectrum of its intermediate, paratenic, and definitive hosts.

Anisakisspp. nematodes are potentially zoonotic parasites; that infects a wide variety of aquatic species worldwide, with marine fish being the paratenic hosts. The aim of study was identify the presence of Anisakidae nematodes, and other parasites in Mugil curema . A total of 96 M . curema obtained from local markets in Tulancingo, Hidalgo, Mexico, were analyzed by necropsy. Only five M . curema present nematode collection in epaxial muscle. The tissues with the highest prevalence of parasites were identified, and samples of epaxial muscle with larval migration analyzed by histopathology. Visible parasites in necropsy tissues were classified according to their morphology. Nematode found in the liver were Contracaecum spp. (41.17%) and Pseudoterranova spp. third stage (7.36%); in the caudal part of the kidney were Anisakis spp. (13.23%), Pseudoterranova spp. third stage (11.77%) and Contracaecum spp. (5.88%); and in epaxial muscle were Anisakis spp. Larva I (5.88%) and Pseudoterranova spp (4.42%). In one fish, Clinostomum spp. was detected in epaxial caudal muscle. The present work reports for the first time the presence of nematodes of the family Anisakidae and Clinostonum spp. metacercariae, with zoonotic potential, in M . curema intended for human consumption in Tulancingo, Hidalgo, Mexico. Resumo Anisakisspp. são parasitas potencialmente zoonóticos que infectam uma grande variedade de espécies aquáticas em todo o mundo, sendo os peixes marinhos hospedeiros paratênicos. O objetivo deste estudo foi identificar a presença de nematóides da família Anisakidae e de outros parasitas em peixes Mugil curema . Um total de 96 M . curema , obtidos em mercados locais em Tulancingo, Hidalgo, México, foram submetidos a necropsia. Apenas cinco M. curema apresentaram coleção de nematóides no músculo epaxial. Os tecidos com maior prevalência de parasitas foram identificados e amostras do músculo epaxial com migração larval foram analisadas por histopatologia. Os nematóides encontrados no fígado foram Contracaecum spp. (41,17%) e Pseudoterranova spp. terceira etapa (7,36%); na parte caudal do rim Anisakis spp. (13,23%), Pseudoterranova spp. terceira etapa (11,77%) e Contracaecum spp. (5,88%); e no músculo epaxial Anisakis spp. larva I (5,88%) e Pseudoterranova spp. (4,42%). Em um peixe, Clinostomum spp. foi detectado no músculo caudal epaxial. O presente trabalho relata pela primeira vez a presença de nematóides da família Anisakidae e Clinostonum spp. metacercárias, com potencial zoonótico, em M. curema destinado ao consumo humano em Tulancingo, Hidalgo, México.

Angiostrongylus cantonensis is the main cause of human eosinophilic meningitis. Humans are accidental hosts, becoming infected due to ingestion of raw intermediate (snails and slugs) or paratenic hosts. Once ingested, the larvae migrate towards the brain where they die, causing the disease. To develop better mollusk control strategies, it is important to first understand what happens in the snail during infection, therefore our purpose was to characterize proteomic, metabolic and immunologic changes in Biomphalaria glabrata 24 h after infection with A. cantonensis. For this purpose, proteins were extracted from infected and uninfected snails and analyzed through mass spectrometry. Hemolymph was also collected, the number of hemocytes was counted and urea, nitric oxide, calcium, glycogen levels as well as alanine and aspartate aminotransferases activities were assessed. The cephalopodal region and gonad-digestive gland complex were dissected and their glycogen content was measured. After infection with A. cantonensis, we observed an increase of hemocytes and granulocytes as well as an increase in hemoglobin type 2 proteins. Temptin-like protein was also found up-regulated in infected snails. Several proteins with structural function (such as myosin heavy chain – striated muscle – like and protein LOC106059779 with ADAM/reprosolin domain) were also differentially expressed, suggesting loss/damage of internal tissues. Increase in phosphoglycerate mutase indicates an increase in glycolysis, possible to compensate the increase in energetic needs. Consequently, there is a decrease in glycogen reserves, particularly in the gonad – digestive gland complex.

American canine hepatozoonosis (ACH) is a debilitating tick-borne disease characterized by pyrexia, body wasting, myopathy, mucopurulent ocular discharge, and periosteal proliferation. The causative agent, Hepatozoon americanum, is an apicomplexan that utilizes the Gulf Coast tick, Amblyomma maculatum, as its definitive host and vector. Unlike most tick-borne disease agents, H. americanum is not transmitted via a tick bite, but is transmitted when canids ingest a tick vector that contains sporulated oocysts within the tick hemocoel or paratenic hosts with cystozoites. Our understanding of H. americanum prevalence is based on its detection in the intermediate host, wild or domestic canids, with domestic canids often showing clinical signs at the time of diagnosis. The frequency of H. americanum in A. maculatum, on the other hand, is unknown; this gap in our knowledge hinders our understanding of transmission risk. Furthermore, current diagnostic assays are limited in efficacy, and serologic assays are not widely available. To begin to address gaps in our knowledge, we developed a TaqMan® multiplex qPCR assay for H. americanum detection in A. maculatum tick extracts and evaluated infection rates in questing adult A. maculatum. Additionally, we used a co-culture system to expose H. americanum stages to host cells for in vitro development. Results from qPCR analysis of over 500 tick extracts revealed no positive samples; this suggests both low transmission risk by adult Gulf Coast tick ingestion in the sampled areas, and that surveillance should be focused in areas where ACH has been diagnosed at higher frequencies. Hepatozoon americanum was detectable by qPCR in co-culture of an infected canine buffy coat with ISE6 (Ixodes scapularis embryonic) tick cells, and microscopic examination of samples from those days revealed some structures that were suspicious for developing stages. These data are a starting point for future work to advance our understanding of H. americanum transmission and mechanisms of disease in canids with ACH.

Parasitic myelopathy caused by Gurltia paralysans in domestic cats is a disease commonly reported in several South American countries. The adult parasite is lodged in the meningeal veins and spinal cord, often causing clinical manifestations of vascular proliferation, thrombophlebitis, and medullary compression. Wild felines are believed to be the definitive hosts of this parasite. The infection occurs through the ingestion of paratenic hosts, but the life cycle of G. paralysans is not yet clearly understood. In this paper, we discuss a case of parasitic myelopathy in a margay (Leopardus wiedii) that died during post-surgical care. Necropsy revealed focal hemorrhages in the thoracolumbar spinal cord. A microscopic examination revealed adult nematodes and eggs inside the veins of subarachnoid space in spinal cord, suggesting G. paralysans infection. This is first description of parasitic myelopathy in a margay in Brazil.

The genus Anisakis represents one of the most widespread groups of ascaridoid nematodes in the marine ecosystem. Three closely related taxa are recognized in the Anisakis simplex (s. l.) complex: A. pegreffii, A. simplex (s. s.) and A. berlandi. They are widely distributed in populations of their intermediate/paratenic hosts (fish and squids) and definitive hosts (cetaceans). A novel nuclear gene locus, metallopeptidase 10 (nas 10) (451 bp), was sequenced and validated on a total of 219 specimens of the three species of Anisakis, collected in fish and cetacean hosts from allopatric areas included in their ranges of distribution. The specimens of Anisakis were first identified by allozymes and sequence analysis of the mtDNA cox2 and EF1α-1 nDNA. The novel nuclear marker has shown fixed alternative nucleotide positions in the three species, i.e. diagnostic at 100%, permitting the species determination of a large number of specimens analyzed in the present study. In addition, primers to be used for amplification-refractory mutation system (ARMS) PCR of the same gene locus were designed at these nucleotide positions. Thus, direct genotyping determination, by double ARMS, was developed and validated on 219 specimens belonging to the three species. Complete concordance was observed between the tetra-primer ARMS-PCR assays and direct sequencing results obtained for the nas 10 gene locus. The novel nuclear diagnostic marker will be useful in future studies on a multi-locus genotyping approach and also to study possible hybridization and/or introgression events occurring between the three species in sympatric areas.Un nouveau marqueur nucléaire et développement d'un test ARMS-PCR ciblant le locus de la métallopeptidase 10 (nas 10) pour identifier les espèces du complexe Anisakis simplex (s. l.) (Nematoda, Anisakidae).Le genre Anisakis représente l’un des groupes de nématodes ascaridoïdes les plus répandus dans l’écosystème marin. Trois taxons étroitement apparentés sont reconnus dans le complexe Anisakis simplex (s. l.) : A. pegreffii, A. simplex (s. s.) et A. berlandi. Ils sont largement répartis dans les populations de leurs hôtes intermédiaires/paraténiques (poissons et calmars) et définitifs (cétacés). Un nouveau locus de gène nucléaire, la métallopeptidase 10 (nas 10) (451 pb), a été séquencé et validé sur un total de 219 spécimens des trois espèces d’Anisakis, collectés chez des hôtes poissons et cétacés de zones allopatriques incluses dans leur aire de répartition. Les échantillons d’Anisakis ont d’abord été identifiés par des allozymes et une analyse des séquences de l’ADNmt cox2 et de l’ADNn EF1α-1. Le nouveau marqueur nucléaire a montré des positions de nucléotides alternatives fixes dans les trois espèces, c’est-à-dire qu’il a permis un diagnostic à 100%, permettant la détermination de l’espèce d’un grand nombre d’échantillons analysés dans la présente étude. De plus, des amorces à utiliser pour la PCR par système de mutation réfractaire à l’amplification (ARMS) du même locus génique ont été conçues à ces positions nucléotidiques. Ainsi, la détermination directe du génotypage, par double ARMS, a été développée et validée sur 219 spécimens appartenant aux trois espèces. Une concordance complète a été observée entre les dosages ARMS PCR tétra-amorces et les résultats de séquençage direct obtenus pour le locus du gène nas 10. Le nouveau marqueur de diagnostic nucléaire sera utile dans les travaux futurs d’une approche de génotypage multi-locus et également pour étudier les éventuels événements d’hybridation et/ou d’introgression se produisant entre les trois espèces dans des zones sympatriques.

Snakes have diverse feeding and living habits, being exposed to a variety of endoparasite communities. However, more studies are still necessary to document these relationships. We examined 18 specimens of the cat-eyed snake Leptodeira annulata from a semi-arid region in Northeast Brazil. Eight taxa of parasites were found, with higher prevalence of cystacanths (Acanthocephala). Five nematode species (Hexametra boddaertii, Oswaldocruzia sp., Oxyascaris sp., Physaloptera sp. and Raillietnema spectans) and the pentastome Raillietiella furcocerca represent a new parasitism record for the host studied. Our results also showed that L. annulata could act as paratenic host for acanthocephalans. These results contribute to the knowledge of the helminth fauna of L. annulata.

Invasive species constitute one of the most serious threats to biodiversity and ecosystems, and they potentially cause economic problems and impact human health. The globally invasive New Guinea flatworm, Platydemus manokwari (Platyhelminthes: Geoplanidae), has been identified as a threat to terrestrial biodiversity, particularly soil-dwelling native species (e.g. molluscs, annelids and other land planarians), and is listed among 100 of the world's worst invasive alien species. We report here, for the first time, P. manokwari occurrences in many locations throughout Thailand, using voluntary digital public participation from the social network portals associated with the Thailand Biodiversity Conservation Group and collections of living flatworm specimens. Mitochondrial cytochrome c oxidase subunit I (COI) sequences confirmed that all collected flatworms were P. manokwari and placed them in the “world haplotype” clade alongside other previously reported specimens from France, Florida (USA), Puerto Rico, Singapore, French Polynesia, New Caledonia, and the Solomon Islands. In addition, infective stage larvae (L3) of the nematode Angiostrongylus malaysiensis were found in the flatworm specimens, with a 12.4% infection rate (15/121 specimens examined). Platydemus manokwari occurrence in Thailand and its capacity to carry L3 of Angiostrongylus should be of concern to biodiversity conservation and human health practitioners, because this invasive flatworm species may be involved in the life cycle of angiostrongylid worms in Thailand.

Angiostrongyliasis is a food-borne parasitic disease caused by the nematode Angiostrongylus cantonensis that can lead to eosinophilic meningitis (EM) or meningoencephalitis in humans. Angiostrongylus cantonensis is prevalent in the Pacific Islands. In recent years, a large number of outbreaks and severe cases have occurred. Several species of mollusk, such as snails and slugs, act as intermediate and paratenic hosts of A. cantonensis. In this study, two cases of EM were found to have been caused by infection with A. cantonensis due to consumption of raw centipedes. To survey the A. cantonensis infections acquired through centipedes that the patients had bought at a vegetable market, we performed etiological examinations and polymerase chain reaction amplification of A. cantonensis genes. Third-instar larvae of A. cantonensis were detected in the centipedes, and specific genes from A. cantonensis were detected in all the specimens. This indicates that the centipede may act as a competent host for the transmission of A. cantonensis. To our knowledge, this is the first report of A. cantonensis infection through the consumption of centipedes.

Toxocariasis is an important, but neglected, worldwide zoonosis. It is considered a primarily soil-transmitted disease, but food-borne transmission has been associated with the consumption either of raw or undercooked meat of paratenic hosts, including birds. Despite the number of experimental studies carried out to evaluate the behavior of Toxocara spp. larvae in birds, their role in the dispersion of eggs into the environment remains unclear. Thus, this study aimed to evaluate the potential of broiler chickens to release Toxocara canis eggs into the environment, and the infectivity of eggs after passage through the intestine. Forty commercial broiler chickens, aged 60 days, were randomly distributed into three groups. Groups 1 (n = 16) and 2 (n = 16) were orally infected with 5000 embryonated and 5000 unembryonated T. canis eggs, respectively. Group 3 (n = 8) served as a control. Following infection, fecal samples from each chicken were examined using a centrifuge-sedimentation technique. At 24-h, 72-h, and 7-day post-infection (PI), four chickens each from the G1 and G2 groups, and two from the G3 group were killed. After euthanasia, the intestinal content and liver were collected for recovery of T. canis larvae. Results revealed that broiler chickens have the potential to disperse both embryonated and unembryonated T. canis eggs, following 2- to 6-h PI. In addition, the eggs shed into the feces of the G2 birds, after incubation in laboratorial conditions, were infective when they were tested in a bioassay using mice. In conclusion, broiler chickens have the potential of dispersing Toxocara spp. eggs into the environment and the eggs passed through the intestine are infective after being incubated in experimental conditions.

Physaloptera spp. are common nematodes found in the stomach and muscles of mammals, reptiles, amphibians, and birds. Physaloptera spp. have a complicated life cycle with multiple definitive hosts, arthropod intermediate hosts, aberrant infections, and possible second intermediate hosts or paratenic hosts. For example, Physaloptera sp. larvae have been found within the tissues of wild northern bobwhite quail (Colinus virginianus), and it is suspected that quail may serve as paratenic or secondary hosts of these parasites. However, because it is not known what role quail play in the life cycle of Physaloptera spp. and descriptions of Physaloptera spp. larvae are limited, molecular tools may be beneficial when identifying these helminths. In this study, we generated primers using universal nematode primers and obtained a partial mitochondrial cytochrome oxidase 1 (COX 1) sequence. Morphological identification of Physaloptera sp. in bobwhite was confirmed via polymerase chain reaction (PCR), and a phylogenetic tree was constructed using the maximum likelihood method. BLAST analysis revealed a strong identity to other Physaloptera spp. and the phylogenetic tree placed all Physaloptera spp. in the same cluster. We also documented a marked increase in Physaloptera infections in bobwhite from 2017 to 2018, and the similarity of these parasites to Onchocerca volvulus and Wuchereria bancrofti may give insight into the increased prevalence we observed. This study demonstrates the usefulness of molecular techniques to confirm the identity of species that may lack adequate descriptions and provides new insight for the diagnosis and potentially overlooked significance of Physaloptera sp. infections of bobwhite in the Rolling Plains ecoregion of Texas.

Larval Baylisascaris nematodes (L3), resulting from transuterine infection and neural migration, were discovered in the cerebrum of sibling moose calves (Alces alces gigas) near 1–3 days in age from Alaska. We provide the first definitive identification, linking morphology, biogeography, and molecular phylogenetics, of Baylisascaris transfuga in naturally infected ungulates. Life history and involvement of paratenic hosts across a broader assemblage of mammals, from rodents to ungulates, in the transmission of B. transfuga remains undefined. Neural infections, debilitating young moose, may seasonally predispose calves to predation by brown bears, facilitating transmission to definitive hosts. Discovery of fatal neurological infections by L3 of B. transfuga in mammalian hosts serves to demonstrate the potential for zoonotic infection, as widely established for B. procyonis, in other regions and where raccoon definitive hosts are abundant. In zones of sympatry for multi-species assemblages of Baylisascaris across the Holarctic region presumptive identification of B. procyonis in cases of neurological larval migrans must be considered with caution. Diagnostics in neural and somatic larval migrans involving species of Baylisascaris in mammalian and other vertebrate hosts should include molecular-based and authoritative identification established in a phylogenetic context., Graphical abstract Image 1, Highlights • Larval Baylisascaris nematodes, demonstrating transuterine infection, were discovered in sibling moose calves from Alaska. • Baylisascaris transfuga was identified through morphology, biogeography and molecular phylogenetics. • Neural infections in ungulate paratenic hosts may predispose calves to predation by brown bears facilitating transmission. • Diagnostics for L3 Baylisascaris in zones of sympatry for multiple species should include molecular-based identification.

Infection with the Rat Lungworm Angiostrongylus cantonensis is the leading cause of human eosinophilic meningoencephalitis worldwide. From its origins in southeastern Asia, the parasite was spread extensively throughout the twentieth century and is now established in many of the world’s warmer regions. Its clinical effects range from mild and transient symptoms, usually headache with peripheral nerve dysfunction, to severe and permanent central nervous system (CNS) damage, occasionally fatal. The severity and prognosis of disease are determined by the larval dose, acquired by ingesting infected intermediate hosts (slugs and snails) or, less often, paratenic hosts, such as crabs, shrimps, frogs, and monitor lizards. Early diagnosis is critical for treatment and depends on clinical suspicion, for laboratory confirmation from blood and cerebrospinal fluid can be delayed and unreliable. Treatment is fraught with difficulty, compounded by conflicting published results. Corticosteroids play a useful role in suppressing early CNS inflammation, but their duration for maintenance becomes problematic in severe infections. Because most of the pathogenesis results from host immuno-inflammatory responses to migrating and dead larvae in the CNS, anthelminthic therapy remains controversial: if effective, it kills viable larvae, arresting them in the CNS and so exacerbating the pathology. In human infections, it is now clear that many larvae do leave the CNS and reach the pulmonary arteries, sometimes with clinical consequences. Pioneering life-cycle studies in rats demonstrated a “subarachnoid phase” in larval development and migration; recent autopsy findings, outlined here, show it also occurs in humans and has some bearing on treatment. One new and four previously reported cases of human infection are analyzed here, with findings indicating that anthelminthic treatment is effective only when given early and should not be commenced beyond 3 weeks after exposure to infection. In endemic areas, treatment should start as soon as this infection is suspected, even without a clear history of exposure, given the unacceptable risks of waiting for diagnostic laboratory confirmation.

Angiostrongyliasis is contracted by the ingestion of or exposure to the zoonotic parasite Angiostrongylus cantonensis, which requires two hosts to complete its life cycle. Rats are known to be the definitive hosts and mollusks are intermediate hosts. It is the intermediate host, or infected paratenic hosts that can pass the infective stage of the disease to humans and other susceptible animals. The purpose of this review is to examine the growing threat of A. cantonensis and its consequences, with particular emphasis on Hawai'i. The increase in disease cases appears to correlate with the migration of the mollusk Parmarion martensi (semi-slug) in Hawai'i. The semi-slug is a very effective host because it carries a heavy burden of the parasite. Rats in Hawai'i also carry heavy parasite burdens. Human diagnosis is difficult due to varied incubation times, multitude of symptoms which can mimic other illnesses, and lack of a low-risk effective diagnostic. Treatment varies with the severity of the symptoms. The introduction and proliferation of the invasive species A. cantonensis, along with a most efficient intermediate host P. martensi, has resulted in an increase in angiostrongyliasis in Hawai'i. In Hawai'i, consumption of local produce is encouraged and the use of rainwater harvesting is a necessity for many on Hawai'i Island, both of which elevate the risk of acquiring angiostrongyliasis. A better understanding of how to best reduce the risk of infection is needed through comprehensive research and community education.

Angiostrongylus vasorum is a parasite that can cause life-threatening disease in dogs. In recent years, A. vasorum has been reported spreading into new areas, in particular towards northeastern Europe. Despite being detected in wildlife in Estonia, no studies have focused on A. vasorum in domestic dogs in the country. We conducted a multi-center study in 2018–2019: at eight small animal clinics, altogether 115 dogs that had clinical signs that could be suggestive of canine angiostrongylosis were enrolled for testing using a commercial A. vasorum antigen detection test. In addition, we collected information on potential risk behaviors for acquiring A. vasorum infection – eating potential intermediate hosts and paratenic hosts – among the dogs, using a questionnaire for dog owners. Moreover, we surveyed knowledge about A. vasorum among veterinarians authorized to work in Estonia. None (0.0%, 95% confidence interval 0.0–2.6) of the 114 dogs included in the study tested A. vasorum antigen positive. Two (2.0%) of the 102 dogs included in the risk behavior analysis had been seen to eat slugs and/or snails, and this behavior was considered possible for further 17 (16.7%) of the dogs. Four (3.9%) of the dogs had been seen to eat frogs, and this behavior was considered possible for further 14 (13.7%) of the dogs. Thirty-eight (90.5%) of the 42 veterinarians who participated in the questionnaire survey selected the correct host and 28 (66.7%) selected the correct vectors of the parasite, from the provided options. Our results indicate that A. vasorum was either not established or not common in dogs in Estonia in 2018–2019, but this needs to be interpreted with caution due to the limited sample size, possible false negative results in recently infected dogs and the limited sensitivity of the test applied. Most local veterinarians seemed to know the parasite, and this awareness and knowledge among veterinarians should be maintained and further improved. Taken the presence of the parasite in local wildlife, it is prudent to include A. vasorum infection in the list of differential diagnoses for dogs with clinical signs that could be suggestive of canine angiostrongylosis in Estonia.

Angiostronglyus vasorum is a cardiopulmonary nematode infecting mainly canids such as dogs (Canis familiaris) and foxes (Vulpes vulpes). Natural infections have also been reported in mustelids and red pandas (Ailurus fulgens fulgens). We report the occurrence of natural A. vasorum infections in a group of captive meerkats (Suricata suricatta), housed at a university facility in Switzerland. A. vasorum first-stage larvae (L1) were initially identified in a pooled faecal sample. Individual samples, investigated with the Baermann-Wetzel technique, revealed that 41% (7/17) of the meerkats were infected, with ranges of 2–125 L1/g faeces. PCR and sequencing of part of the ITS-2 region resulted in 100% identity with A. vasorum. Infected animals did not show clinical signs. One meerkat died two days after diagnosis. Upon necropsy one adult specimen was recovered; histological examination of the lung revealed granulomatous pneumonia caused by A. vasorum larvae and eggs as well as intima and media hyperplasia and isolated arteriosclerosis of larger lung vessels. However, the cause of death was a spleen rupture with associated blood loss. All meerkats were topically treated with 10 mg imidacloprid/2.5 mg moxidectin per animal, after which they became negative in all follow up faecal examinations. Potential intermediate (gastropods) and paratenic hosts (birds) were collected from within or outside the meerkats enclosure. Gastropods were examined by PCR and bird samples by digestion. Four out of 193 (2.1%) gastropod samples were positive for A. vasorum, whereas none of the bird samples were positive. Meerkats, belonging to the Herpestidae, therefore are suitable definitive hosts for A. vasorum, with production and excretion of live L1. Meerkats kept in captivity in areas where A. vasorum is endemic and with potential contact to intermediate hosts are at risk of infection. Regular faecal examinations including Baermann-Wetzel technique should be considered., Graphical abstract Image 1, Highlights • Meerkats are suitable definitive hosts for Angiostrongylus vasorum. • This is the first case of A. vasorum in the Herpestidae family. • A. vasorum larvae and eggs caused granulomatous pneumonia in an infected meerkat. • 2% of gastropods around the meerkats enclosure were positive for A. vasorum L3. • Captive animal species in areas where A. vasorum is endemic are at risk of infection.