DESCRIPTION OF FEMALE DENDRITOBILHARZIA PULVERULENTA (BRAUN, 1901) SKRJABIN, 1924 FROM TWO NEW AVIAN HOSTS IN NAMIBIA WITH PHYLOGENETIC ANALYSES AND COMMENTS ON SEVERAL TAXONOMICALLY UNCERTAIN AVIAN SCHISTOSOME SEQUENCES.

During a 2021 parasitological survey of birds in the Nyae Nyae-Khaudum Dispersal Area (Kavango-Zambezi Transfrontier Conservation Area, Namibia), we collected 9 specimens of Dendritobilharzia pulverulenta (Braun, 1901) Skrjabin, 1924 infecting the blood (heart lumen) of a white-backed duck, Thalassornis leuconotus (Eyton, 1838) (Anseriformes: Anatidae), and a fulvous whistling duck, Dendrocygna bicolor (Vieillot, 1816) (Anatidae). These flukes were fixed for morphology and preserved for DNA extraction. We assigned our specimens to DendritobilharziaSkrjabin and Zakharow, 1920 because they were strongly dorso-ventrally flattened in both sexes and had an intestinal cyclocoel with a zig-zag common cecum with lateral dendritic ramifications, numerous testes posterior to the cyclocoel and flanking the dendritic ramifications, and a tightly compacted convoluted ovary as well as lacking an oral sucker, ventral sucker, and gynaecophoric canal. Further, our specimens were morphologically identical to previously published descriptions of D. pulverulenta. Sequences of the 28S from our specimens were nearly identical to those identified as D. pulverulenta from North America (New Mexico), and our 28S phylogenetic analysis recovered D. pulverulenta within a polytomy of other Gigantobilharziinae spp. The CO1 phylogenetic analysis recovered a monophyletic Dendritobilharzia and, with low taxon sampling, a monophyletic Gigantobilharzia. This is the first record of a species of Dendritobilharzia infecting these ducks as well as the first record of an adult Dendritobilharzia from sub-Saharan Africa. The original description of adult D. pulverulenta (type locality: northern Sudan) was based on 2 males only, and hence the present study is the first description of female D. pulverulenta from Africa (the continent of the type locality). We reassign 2 Gigantobilharziinae spp. based on morphology and nucleotide evidence: Gigantobilharzia ensenadense (Lorenti, Brant, Gilardoni, Diaz, and Cremonte, 2022) Dutton and Bullard, n. comb., and Gigantobilharzia patagonense (Lorenti, Brant, Gilardoni, Diaz, and Cremonte, 2022) Dutton and Bullard, n. comb. We also comment on several avian schistosome sequences whose identities need confirmation or that likely have been misidentified.

Expanding the swimmer's itch pool of the Benelux: a first record of the neurotropic Trichobilharzia regenti and potential link to human infection.

BACKGROUND: Swimmer's itch, an allergic contact dermatitis caused by avian and mammalian blood flukes, is a parasitic infection affecting people worldwide. In particular, avian blood flukes of the genus Trichobilharzia are infamous for their role in swimmer's itch cases. These parasites infect waterfowl as a final host, but incidental infections by cercariae in humans are frequently reported. Upon accidental infections of humans, parasite larvae will be recognized by the immune system and destroyed, leading to painful itchy skin lesions. However, one species, Trichobilharzia regenti, can escape this response in experimental animals and reach the spinal cord, causing neuroinflammation. In the last few decades, there has been an increase in case reports across Europe, making it an emerging zoonosis. METHODS: Following a reported case of swimmer's itch in Kampenhout in 2022 (Belgium), the transmission site consisting of a private pond and an adjacent creek was investigated through a malacological and parasitological survey. RESULTS: Six snail species were collected, including the widespread Ampullaceana balthica, a well-known intermediate host for Trichobilharzia parasites. Shedding experiments followed by DNA barcoding revealed a single snail specimen to be infected with T. regenti, a new species record for Belgium and by extension the Benelux. Moreover, it is the most compelling case to date of the link between this neurotropic parasite and cercarial dermatitis. Additionally, an Echinostomatidae sp. and Notocotylus sp. were isolated from two other specimens of A. balthica. However, the lack of reference DNA sequences for these groups in the online repositories prevented genus- and species-level identification, respectively. CONCLUSIONS: The presence of T. regenti in Belgium might have severe clinical implications and its finding highlights the need for increased vigilance and diagnostic awareness among medical professionals. The lack of species-level identification of the other two parasite species showcases the barcoding void for trematodes. Overall, these findings demonstrate the need for a Belgian framework to rapidly detect and monitor zoonotic outbreaks of trematode parasites within the One Health context.

VECTOR COMPATIBILITY OF NEW MEXICO GALBA SPECIES WITH THE CANINE SCHISTOSOME HETEROBILHARZIA AMERICANA, INCLUDING THE FIRST REPORT OF GALBA SCHIRAZENSIS AS A COMPATIBLE HOST.

The indigenous North American mammalian schistosome Heterobilharzia americana has recently attracted attention for causing outbreaks in dogs in states outside of its southeastern U.S. distribution. Although H. americana has yet to be reported in New Mexico, we examined 2 New Mexico isolates of Galba snails to determine their susceptibility to experimental infection with an isolate of H. americana from Utah. One of the Galba isolates from the Rio Grande bosque in the Albuquerque suburb of Corrales was identified as Galba humilis, and like specimens of the same taxon from Utah, proved susceptible to H. americana (27.6% of exposed surviving snails positive). The second Galba isolate sourced from the northern mountains of New Mexico, which surprisingly was revealed to be Galba schirazensis based on cytochrome c oxidase 1, 16S rRNA, and the internal transcribed spacer 2 markers, was also susceptible to H. americana (56.3% of exposed surviving field-derived snails and 46.4% first generation [F1] snails positive). This is the first report of the latter snail being a compatible snail host for H. americana. As G. schirazensis has a wide, albeit spotty, distribution and is considered an invasive species, it provides yet another opportunity for H. americana to expand its known range, potentially including the state of New Mexico as well.

Diversity of trematode cercariae among naturally infected lymnaeid snails from Phayao, Thailand.

Lymnaeids are aquatic snails playing an important role in the transmission of many parasitic trematode species of veterinary and medical importance. In this study, we assessed the presence of cercarial flukes in naturally infected lymnaeid snails from Phayao province, Thailand, and determined the species diversity of both the intermediate snail hosts and parasite larvae. A total of 3,185 lymnaeid snails were collected from paddy fields at 31 sites in eight districts of Phayao province between October 2021 and December 2022. Larval fluke infection was assessed using the cercarial shedding method. The collected snails as well as emerging cercariae were identified at the species level via morphological and molecular methods. The sequences of snail internal transcribed spacer region 2 (ITS2) and cercarial 28S ribosomal RNA gene (28S rDNA) and cytochrome C oxidase1 (Cox1) were determined by PCR amplification and sequencing. Three species of lymnaeid snails were detected in this study, including Radix (Lymnaea) rubiginosa (Michelin, 1831), Radix (Lymnaea) swinhoei (Adams, 1866) and Austropeplea viridis (Quoy & Gaimard, 1832), of which R. rubiginosa was the most abundant, followed by A. viridis and R. swinhoei. The overall rate of trematode cercarial infection in the lymnaeid snails was 2.8% (90/3,185); the cercarial infection rate in R. rubiginosa and A. viridis was 3.5% (60/1,735) and 3.1% (30/981), respectively. No larval fluke infection was observed in the studied R. swinhoei (0/469). Nine morphotypes of cercariae were detected at 15 sites from four districts. The emerging cercariae were molecularly identified as Clinostomum sp., Aporocotylidae sp., Apharyngostrigea sp., Trichobilharzia sp., Apatemon sp., Pegosomum sp., Petasiger sp., Echinostoma revolutum and Plagiorchis sp. These findings emphasize the occurrence and diversity of trematode cercariae among naturally infected lymnaeid snails in Phayao province and could contribute to broadening our understanding of the host-parasite relationships between trematodes and their first intermediate hosts as well as developing effective interventions to control trematode parasitic diseases.

Phylogenetic analysis of avian schistosome Trichobilharzia regenti (Schistosomatidae, Digenea) from naturally infected hosts in northern Iran.

BACKGROUND: Trichobilharzia regenti (T. regenti) is an avian schistosomatid fluke species that causes human cercarial dermatitis (HCD) in areas of aquaculture in northern Iran. Understanding the phylogenetic relationships and genetic diversity of this thread-like fluke will deepen our thoughtful of avian schistosomiasis epidemiology and lead to more effective HCD control in the region. OBJECTIVES: To determine the life cycle of nasal Trichobilharzia in aquatic birds as well as aquatic snails and also identify the haplotype diversity of the isolates in Mazandaran Province, northern Iran. METHODS: In the present study, adult or egg of Trichobilharzia isolated from aquatic birds as well as schistosomes cercariae isolated from aquatic snails in Mazandaran Province, northern Iran, belonged to the authors' previous research, were examined. Molecular studies and phylogenetic analysis were carried out on these schistosomes samples. RESULTS: The phylogenetic analysis of the ITS1 and COX1 genes in isolated schistosomes revealed that all samples belong to the T. regenti clade. Remarkably, based on phylogenetic results, these schistosomes samples from Anas platyrhynchos domesticus, A. platyrhynchos, Spatula clypeata and Lymnaea stagnalis grouped together with previously sequenced samples from Iran (Trichobilharzia cf. regenti). Unlike the phylogenetic tree and haplotype network of COX1 gene, ITS1 did not show distinct clusters. CONCLUSION: This study completed the puzzle of the disease in Mazandaran Province by isolating and genotyping furkocercariae from L. stagnalis that was consistent with the isolated new genotype from ducks. For the first time in Iran, this confirmed the potential role of L. stagnalis snails in the transmission of the disease.

A report of Bilharziella polonica cercariae in Knowsley Safari, Prescot, United Kingdom, with notes on other trematodes implicated in human cercarial dermatitis.

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.

Schistosomes in the Persian Gulf: novel molecular data, host associations, and life-cycle elucidations.

Avian schistosomes, comprise a diverse and widespread group of trematodes known for their surprising ability to switch into new hosts and habitats. Despite the considerable research attention on avian schistosomes as causatives of the human cercarial dermatitis, less it is known about the diversity, geographical range and host associations of the marine representatives. Our molecular analyses inferred from cox1 and 28S DNA sequence data revealed presence of two schistosome species, Ornithobilharzia canaliculata (Rudolphi, 1819) Odhner, 1912 and a putative new species of Austrobilharzia Johnston, 1917. Molecular elucidation of the life-cycle of O. canaliculata was achieved for the first time via matching novel and published sequence data from adult and larval stages. This is the first record of Ornithobilharzia from the Persian Gulf and globally the first record of this genus in a potamidid snail host. Our study provides: (i) new host and distribution records for major etiological agents of cercarial dermatitis and contributes important information on host-parasite relationships; (ii) highlights the importance of the molecular systematics in the assessment of schistosome diversity; and (iii) calls for further surveys to reach a better understanding of the schistosome diversity and patterns of relationships among them, host associations, transmission strategies and distribution coverage.

Diversity of Trichobilharzia in New Zealand with a new species and a redescription, and their likely contribution to cercarial dermatitis.

In response to annual outbreaks of human cercarial dermatitis (HCD) in Lake Wanaka, New Zealand, ducks and snails were collected and screened for avian schistosomes. During the survey from 2009 to 2017, four species of Trichobilharzia were recovered. Specimens were examined both morphologically and genetically. Trichobilharzia querquedulae, a species known from four continents, was found in the visceral veins of the duck Spatula rhynchotis but the snail host remains unknown. Cercaria longicauda [i.e. Trichobilharzia longicauda (Macfarlane, 1944) Davis, 2006], considered the major aetiological agent of HCD in Lake Wanaka, was discovered, and redescribed from adults in the visceral veins of the duck Aythya novaeseelandiae and cercariae from the snail Austropeplea tomentosa. Recovered from the nasal mucosa of Ay. novaeseelandiae is a new species of Trichobilharzia that was also found to cycle naturally through Au. tomentosa. Cercariae of a fourth species of Trichobilharzia were found in Au. tomentosa but the species remains unidentified.

Heterobilharzia americana infection in dogs: A retrospective study of 60 cases (2010-2019).

BACKGROUND: The trematode Heterobilharzia americana (HA) causes granulomatous gastrointestinal and hepatic disease in dogs. Before 2008, diagnosis relied on saline fecal sedimentation or histopathology, and earlier reports primarily described dogs with advanced disease or cases diagnosed incidentally at necropsy. The advent of a fecal PCR test has facilitated the diagnosis of HA and provided insights into manifestations and response to treatment. OBJECTIVES: Describe the clinical findings, response to treatment, and outcome for dogs infected with HA. ANIMALS: Sixty dogs diagnosed with HA between 2010 and 2019. METHODS: Retrospective study. Medical records were searched for dogs diagnosed with HA by fecal PCR testing, identification of ova in feces, or histopathology. RESULTS: Mean age was 7.5 (±4.1) years and weight was 23.2 (±10.18) kg. Clinical signs included diarrhea (55.8%), vomiting (46.2%), and weight loss with or without anorexia (15.4%). Laboratory abnormalities included hyperglobulinemia (42.6%) and increased liver enzyme activities (30%). More than 40% of dogs had an eosinophil count >500/µL. Hypercalcemia attributable to HA was identified in only 4 dogs. Pinpoint hyperechoic foci were noted in intestines, liver, or mesenteric lymph nodes during transabdominal ultrasonography in 64.4% of dogs. Survival data was available for 34 dogs, of which 73.5% (25) were alive 6 months after diagnosis. CONCLUSIONS AND CLINICAL IMPORTANCE: Hyperglobulinemia, high eosinophil count, and ultrasonographic evidence of visceral mineralization were suggestive of infection. Hypercalcemia was uncommon. Combination treatment with praziquantel and fenbendazole was variably effective, and 17.6% of treated dogs with known outcome died as a result of HA infection.

Non-coding Regions of Mitochondrial DNA and the cox1 Gene Reveal Genetic Variability Among Local Belarusian Populations of the Causative Agent of Cercarial Dermatitis, Bird Schistosome Trichobilharzia szidati (Digenea: Schistosomatidae).

INTRODUCTION: The cercariae of avian blood flukes Trichobilharzia szidati (Digenea, Schistosomatidae) are known to cause cercarial allergic dermatitis ("swimmer's itch") in humans. Global epidemics can have significant impacts on local tourism-related economies in recreational areas. Little is known about the genetic polymorphism of the parasite population, or about the variability of the non-coding regions of mitochondrial DNA (mtDNA) and the possibility of using this as a genetic marker. MATERIALS AND METHODS: The T. szidati cercariae were collected over 7 years from 33 naturally infected Lymnaea stagnalis snails from five sites at two neighboring lakes in Belarus. We investigated the variability of the short (SNR) and long (LNR) non-coding regions of mt DNA and the genetic diversity within the 1125-bp sequences of the gene for subunit 1 of cytochrome c oxidase (cox1). RESULTS: In the SNR sequences, we found only length variability caused by changes in the number of bases in the mononucleotide tracts T6-T8. LNR demonstrates high variability in nucleotide sequence length (182-260 bp) depending on the presence of two long deletions of 59 and 78 nucleotides. Both mitochondrial loci (LNR and cox1) are characterized by high haplotype diversity (H = 0.922 and H = 1.0, respectively); the nucleotide diversity is significantly higher for LNR (π = 1.926 ± 0.443) compared to cox1 (π = 0.704 ± 0.059). Phylogenetic reconstructions based on the variability of each of the loci (LNR and cox1) and their concatenated sequences revealed their shallow structure and the absence of a correlation between the distribution of single-nucleotide polymorphisms and the geographic origin of parasites from two Belarusian lakes. We identified at last four weakly sublineages in the phylogenetic pattern of T. szidati. The carriers of each deletion have specific patterns for each of the two loci and form their own phylogeographic sublineages. An association between two fixed LNR substitutions and a fixed non-synonymous substitution in cox1 was found in four representatives of one lineage that had a short deletion in the LNR. CONCLUSIONS: This study clarified the phylogeographic structure of the Belarusian population of T. szidati. Our data provide the basis for the use two mt markers in large-scale population studies of the parasite, as well as for studying the molecular evolution of coding and non-coding mtDNA in trematodes.

Heterobilharzia americana infection in a Grant's zebra (Equus burchelli boehmi).

Heterobilharzia americana is a trematode parasite (family: Schistosomatidae) of mammalian wildlife that occasionally infects domestic dogs and horses in the southeastern United States. This report presents the first case of H. americana infection in a Grant's zebra (Equus burchelli boehmi). The parasite was diagnosed post mortem as an incidental finding by histopathologic identification of H. americana eggs in the liver parenchyma after the zebra succumbed in an accidental traumatic death. Molecular analysis of a portion of the parasite small subunit ribosomal RNA gene demonstrated 100% identity with DNA sequences from parasites previously characterized in domestic horses. Equine infections with H. americana are considered uncommon, but are probably underdiagnosed.

A new schistosome (Digenea: Schistosomatidae) from the nasal tissue of South America black-necked swans, Cygnus melancoryphus (Anatidae) and the endemic pulmonate snail Chilina gibbosa.

To date, 9 species of Schistosomatidae have been found parasitizing the nasal tissues of mammal and bird hosts in the Eastern Hemisphere, 5 species in Rwanda (Africa), 2 in Australia (Oceania) and 2 in Eurasia. During a parasitological survey of black necked swans, Cygnus melancoryphus, an anatid endemic to South America, schistosome worms in the nasal tissue were found; the first in the Americas. Morphological results based on male worms and in isolated eggs. The worms have a spiny tegument, filiform body with rounded posterior end, two muscular suckers, a robust gynaecophoric channel with thickened cross bands, and around 130 testes. The eggs are elongate with an asymmetrical bulge, with a slender process at one end and a longer curved process at the other. Diagnostic morphological characteristics do not match with any schistosome genus. Part of the mitochondrial cox1 and nuclear DNA 28S partial genes were sequenced and compared to Schistosomatidae in GenBank. The genetic results confirm the distinctiveness of the specimens since they do not group with any described genus or undescribed lineage other than cercariae of "Chilina lineage 1" that emerge from the Patagonian Chilina gibbosa, a freshwater snail endemic to South America. Based on morphological and genetic characterization of these schistosomes, these specimens represent a new genus and species that parasitizes black necked swans as adults in the nasal tissue, and C. gibbosa is the first intermediate host, both hosts being endemic to South America.

Dermatological and Molecular Evidence of Human Cercarial Dermatitis in North-Eastern Poland.

Swimmer's itch or human cercarial dermatitis (HCD) appears as a skin rash caused by an allergic reaction to larval (cercariae) flatworm parasites of the family Schistosomatidae. In our study, two cases of HCD were analyzed; both of them were reported in people swimming in Lake Pluszne. In the summer of 2018, a sample of 397 snails was collected at swimming sites in that area. Five Lymnaea stagnalis (1.9%) were found to host cercariae of bird schistosomes. Positive samples were selected by amplification of the Internal Transcribed Spacers (ITS) gene region. Sequence analysis confirmed that they were homologous with European isolates of Trichobilharzia szidati. The cases reported in this article are the first confirmed cases of HCD in this lake. This study demonstrates that there is a rationale for conducting screening studies of regions with a high recreational potential.

Molecular evidence of new freshwater turtle blood flukes (Digenea: Spirorchiidae) in the intermediate snail host Biomphalaria occidentalis Paraense, 1981 in an urban aquatic ecosystem in Brazil.

In this study, two potentially new species of turtle blood flukes (TBFs) (Digenea: Spirorchiidae) have been recorded from South Brazil. The spirorchiid parasites infect the vascular system of turtles, thereby compromising their health. The life cycle of these parasites is not well studied. The larval stage of cercaria is found in intermediate gastropod hosts, with some species presenting similar morphological characteristics, which can result in misinterpretations when using only morphological taxonomy for species identification. In this study, we recorded a single morphotype belonging to the family Spirorchiidae in Biomphalaria occidentalis in an urban aquatic ecosystem in Brazil. However, molecular data (28S rDNA and cytochrome oxidase subunit I) confirmed the presence of two species of Spirorchiidae in the sampled environment; both phylogenetically close to genera previously studied in freshwater turtles from the Peruvian Amazon. In this study, species characterization was possible because of molecular tools. We recommend using more than one molecular marker in future studies focusing on TBFs, which need attention about their evolutionary history and ecology to understand their distribution in South America.

First confirmed record of Trichobilharzia franki Müller & Kimmig, 1994, from Radix auricularia (Linnaeus, 1758) for Austria.

Avian schistosomes are of medical and veterinary importance as they are responsible for the annually occurring cercarial dermatitis outbreaks. For Austria, so far, only Trichobilharzia szidati Neuhaus 1952 was confirmed on species level as causative agent of cercarial dermatitis. Here we present the first record of Trichobilharzia franki Müller & Kimmig 1994 in Austria. The species was detected during a survey of digenean trematodes in Upper Austrian water bodies. Furthermore, we provide DNA barcodes of T. franki as well as measurements of several parasite individuals to indicate the intraspecific diversity. We also recommend the usage of an alternative primer pair, since the "standard COI primer pair" previously used for Schistosomatidae amplified an aberrant fragment in the sequence of T. franki. Overall, our study shows how limited our knowledge about occurrence and distribution of avian schistosomes in Austria is and how important it is to acquire such a knowledge to estimate ecological and epidemiological risks in the future.

A new species and emendation of the seldom reported Enterohaematotrema Mehra, 1940 (Digenea: Schistosomatoidea), including a revised phylogenetic hypothesis for turtle blood flukes.

Enterohaematotrema Mehra, 1940 is emended herein based upon a review of the literature and a description of a new species (Enterohaematotrema triettruongi n. sp.) infecting yellow-headed temple turtles, Heosemys annandalii (Boulenger) (Cryptodira: Geoemydidae), in the Mekong River, Vietnam. The new species differs from the published descriptions of its congeners Enterohaematotrema palaeorticum Mehra, 1940 and Enterohaematotrema hepaticum (Simha, 1958) Simha & Chattopadhyaya, 1980 by having two distinctive oesophageal glands, a short and eversible cirrus (vs protrusive with 3 distinct processes), a dorsal common genital pore that is sinistral (vs ventral and medial), a transverse (vs longitudinal) external seminal vesicle, an oviducal seminal receptacle that is sinistral (vs dextral), and a vitellarium distributing from the caecal bifurcation (anterior to the ventral sucker) to the caecal tips (vs vitellarium not extending anteriad beyond ventral sucker in E. palaeorticum or vitellarium wholly posterior to the terminal genitalia in E. hepaticum). A phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) recovered Enterohaematotrema and Platt Roberts & Bullard, 2016 as sister taxa that share a recent common ancestor with the clade comprising Ruavermis Dutton & Bullard, 2020 and Coeuritrema Mehra, 1933. These flukes collectively comprise a monophyletic group of southeast Asian turtle blood flukes. This analysis also indicated that the massive, longitudinal metraterm of species of Enterohaematotrema and Uterotrema Platt & Pichelin, 1994 represents homoplasy (convergent evolution). The present study comprises the first morphological study of original specimens of any species of Enterohematotrema in more than 50 years and is the first molecular phylogenetic placement of the genus among the various turtle blood fluke lineages.

Confirmation of the presence of zoonotic Trichobilharzia franki following a human cercarial dermatitis outbreak in recreational water in Slovakia.

Human cercarial dermatitis is a parasitic disease that causes an allergic reaction in the skin (swimmer's itch) as a consequence of contact with cercariae of bird schistosomes present in water, mainly of the genus Trichobilharzia Skrjabin et Zakarow, 1920. The main objective of the study was to confirm the presence of the zoonotic disease agent following reports of human infections in recreational water in Slovakia. We identified two species of freshwater snails at Kosice Lake, Radix auricularia (Linnaeus, 1758) and Physa acuta (Draparnaud, 1805). Trematode infections were observed only in R. auricularia. Of the 62 snails collected, 11 (17.7%) were infected with 5 different species of larval stages of trematodes. The blood fluke Trichobilharzia franki was found in 2 (3.2%) of the examined snails. The present record provides the first evidence that T. franki from the pulmonate snail R. auricularia represents a source of human cercarial dermatitis in recreational water in Slovakia. Our finding complements the easternmost records of both swimmer's itch and the confirmed occurrence of a bird schistosome in a waterbody in Europe. The present work suggests that the health risks associated with trichobilharziasis need to be further studied by detailed monitoring of the occurrence of the major causative agent of human cercarial dermatitis, T. franki.

Report of high prevalence of schistosome infection in Biomphalaria snails from a geographic area with no previous prevalence of human schistosomiasis in Nigeria.

Previous study using the traditional method of screening snails for infection reported shedding of Schistosoma cercaria by Biomphalaria snails from a river in Nkalagu, southeastern Nigeria. This is contrary to published reports that Biomphalaria from this part of the country does not shed schistosome cercaria. Here, we employed the use of polymerase chain reaction (PCR) methods to screen and characterize the Biomphalaria snails from Nkalagu. Snails were collected from the River Uzuru in dry season, identified and subjected to molecular assays. Genomic DNA (gDNA) was extracted from whole tissues of the 212 Biomphalaria snails and amplified using conventional PCR to check for the schistosome infection level. Assay for the detection of S. mansoni infection was further done using a nested PCR (nPCR). We amplified the entire internal transcribed spacer 2 (ITS2) regions from gDNA of the 212 snails. The representative samples were sequenced and subjected to BLAST searches to confirm snail species. Of the 212 snails screened, 164 (77.4%) of the snails were infected with schistosomes, but only 16 (9.76%) of the snails were positive for S. mansoni infection. Amplification of the snails' ITS2 region yielded a product of 460 bp, and BLAST searches confirmed the snails to be B. pfeifferi, and BLAST searches confirmed the snails to be B. pfeifferi. This paper reports for the first time the presence of S. mansoni positive B. pfeifferi in Nkalagu, which suggest there may be cases of intestinal schistosomiasis in this part of Nigeria.

The repositioning of epigenetic probes/inhibitors identifies new anti-schistosomal lead compounds and chemotherapeutic targets.

BACKGROUND: Praziquantel represents the frontline chemotherapy used to treat schistosomiasis, a neglected tropical disease (NTD) caused by infection with macro-parasitic blood fluke schistosomes. While this drug is safe, its inability to kill all schistosome lifecycle stages within the human host often requires repeat treatments. This limitation, amongst others, has led to the search for novel anti-schistosome replacement or combinatorial chemotherapies. Here, we describe a repositioning strategy to assess the anthelmintic activity of epigenetic probes/inhibitors obtained from the Structural Genomics Consortium. METHODOLOGY/PRINCIPLE FINDINGS: Thirty-seven epigenetic probes/inhibitors targeting histone readers, writers and erasers were initially screened against Schistosoma mansoni schistosomula using the high-throughput Roboworm platform. At 10 µM, 14 of these 37 compounds (38%) negatively affected schistosomula motility and phenotype after 72 hours of continuous co-incubation. Subsequent dose-response titrations against schistosomula and adult worms revealed epigenetic probes targeting one reader (NVS-CECR2-1), one writer (LLY-507 and BAY-598) and one eraser (GSK-J4) to be particularly active. As LLY-507/BAY-598 (SMYD2 histone methyltransferase inhibitors) and GSK-J4 (a JMJD3 histone demethylase inhibitor) regulate an epigenetic process (protein methylation) known to be critical for schistosome development, further characterisation of these compounds/putative targets was performed. RNA interference (RNAi) of one putative LLY-507/BAY-598 S. mansoni target (Smp_000700) in adult worms replicated the compound-mediated motility and egg production defects. Furthermore, H3K36me2, a known product catalysed by SMYD2 activity, was also reduced by LLY-507 (25%), BAY-598 (23%) and siSmp_000700 (15%) treatment of adult worms. Oviposition and packaging of vitelline cells into in vitro laid eggs was also significantly affected by GSK-J4 (putative cell permeable prodrug inhibitor of Smp_034000), but not by the related structural analogue GSK-J1 (cell impermeable inhibitor). CONCLUSION/SIGNIFICANCE: Collectively, these results provide further support for the development of next-generation drugs targeting schistosome epigenetic pathway components. In particular, the progression of histone methylation/demethylation modulators presents a tractable strategy for anti-schistosomal control.

Heads or tails? Differential translational regulation in cercarial heads and tails of schistosome worms.

Schistosomes are obligate helminths responsible for over 218 million cases of human schistosomiasis in 78 countries around the world. Infection occurs when free-swimming cercariae penetrate human skin and initiate developmental progression into parasitic obligate worms that consume red blood cells. Transcriptomic studies of infectious cercariae reveal abundant mRNAs associated with energy metabolism and host invasion. However, the cercaria is mostly transcriptionally quiescent, suggesting that most mRNAs are primed prior to cercarial escape from the snail host. The use of transcriptomics to understand protein expression presumes that transcription and translation are functionally coupled and the cercarial stage has categorically been treated as a single unit for -omic analysis. Per contra, the relationship between transcription and translation in infectious cercariae has not been described. To understand the correlation between transcription and translation in cercariae, we separately measured nascent translation levels in cercarial heads, cercarial tails and in the developing schistosomula, the next stage of its life cycle. The loss of the cercarial tail is essential for the transformation from a cercaria to a schistosomulum. We observed that translation was initially limited and the translation rate accelerated during the first 72-hours after tail loss. When we tested nascent translation in cercarial heads, cercarial tails, whole cercariae, and 4-hour schistosomula, we found that translation is significantly upregulated in the cercarial tail when compared to the cercarial head and that translation was undetectable in heads using immunofluorescent image quantification (p = .0005). These data represent a major shift in how we understand the cercarial stage. The cercarial head is mostly transcriptionally and translationally quiescent while being sufficient for progression into a schistosomulum. In addition, transcription and translation are not linked in Schistosoma mansoni cercaria. Thus, our current conceptual approach of treating the cercaria as a single functional unit for -omic studies may be insufficient to understand cercarial development.