Cercarial dermatitis (CD; swimmer's itch) is a re-emerging skin disease caused by avian schistosomes, including Trichobilharzia franki. Here, we present morphological, genetic, and experimental evidence confirming the involvement of T. franki in recent CD outbreaks across Czechia. Ocellate furcocercariae were collected from Radix auricularia at four sites and identified as T. franki through ITS1 sequencing. Despite minor morphological differences from previously reported specimens, all isolates belonged to the genetically uniform T. franki "auricularia" clade. Experimental infection of mice with T. franki resulted in a ∼ 60 % penetration rate, accompanied by early-onset scratching and transient weight loss. Gross pathology demonstrated hemorrhages on lung surfaces and splenic atrophy at 2 days post-infection (dpi), along with a prominent enlargement of parotid lymph nodes at both 2 and 7 dpi. Histological examination of the skin revealed viable schistosomula, moderate leukocyte infiltration, epidermal hyperplasia, and the formation of hyperkeratotic crusts at 2 dpi. By 7 dpi, parasites were no longer detectable, but epidermal pathology persisted. In the lungs, eosinophil-rich foci and multifocal hemorrhages were observed at 2 dpi, transitioning to neutrophil-dominated lesions at 7 dpi, despite the absence of detectable schistosomula. Splenocytes from infected mice responded to homologous and heterologous cercarial antigens by producing IFN gamma, IL-4, and IL-10, indicating a mixed Th1/Th2/Treg profile and notable species cross-reactivity. However, parasite-specific IgG remained undetectable at 7 dpi. These findings confirm T. franki as the causative agent of CD outbreaks and underscore its capacity to induce localized and systemic pathology and immune response, cross-reacting with other schistosomes.

• Keywords
• Avian schistosomes, Cercarial dermatitis, Lungs, Skin, Trichobilharzia franki,
• Publication type
• Journal Article MeSH

BACKGROUND: Cercarial dermatitis (CD), or swimmer's itch, is a water-borne allergic skin reaction caused by the penetration of the larval stages of bird schistosomes (cercariae) into the skin. Members of the genus Trichobilharzia are the primary causative agents of CD worldwide. Due to the increasing number of cases, CD is regarded as a (re)emerging disease. Outbreaks in recreational waters can significantly impact public health and local economies. Environmental monitoring of Trichobilharzia is crucial for outbreak prediction and public health management. However, conventional methods, such as cercarial shedding and snail dissections, are labour-intensive and lack sensitivity. To overcome these limitations, we present a molecular toolkit that combines loop-mediated isothermal amplification (LAMP), quantitative polymerase chain reaction (qPCR), and multiplex PCR for rapid, sensitive, and accurate detection and identification of Trichobilharzia spp. from various biological samples. METHODS: Tricho-LAMP and Tricho-qPCR were designed and optimised for Trichobilharzia DNA detection. A multiplex PCR assay was also developed and optimised to identify the three main species causing CD in Europe (Trichobilharzia franki, T. szidati, and T. regenti). RESULTS: Tricho-LAMP specifically detected T. regenti and T. franki at 10-3 ng, and T. szidati at 10-2 ng per reaction with genomic DNA. Using gBlocks synthetic DNA, Tricho-LAMP achieved 100% amplification at 10,000 copies and 85% amplification at 1000 copies, with decreasing success at lower concentrations. Tricho-qPCR showed the highest sensitivity, detecting all species down to 10-4 ng per reaction and showing a limit of detection at 10 copies of synthetic DNA in the reaction. Multiplex PCR allowed reliable species differentiation via gel electrophoresis of the PCR products, but the assay had the lowest sensitivity. CONCLUSIONS: We provide a molecular toolkit consisting of LAMP, qPCR, and multiplex PCR. By exhibiting high sensitivity, Tricho-LAMP and Tricho-qPCR assays are potentially suitable for environmental DNA (eDNA)-based environmental monitoring of bird schistosomes, by both researchers and public health authorities. Multiplex PCR can be used for species determination without the need for further sequencing.

• Keywords
• Trichobilharzia, Bird schistosomes, Cercarial dermatitis, Detection, LAMP, Monitoring, Multiplex PCR, qPCR,
• MeSH
• Molecular Diagnostic Techniques * methods   MeSH
• DNA, Helminth genetics   MeSH
• Snails parasitology   MeSH
• Trematode Infections * diagnosis   parasitology   veterinary   MeSH
• Real-Time Polymerase Chain Reaction * methods   MeSH
• Multiplex Polymerase Chain Reaction * methods   MeSH
• Birds parasitology   MeSH
• Schistosomatidae * genetics   isolation & purification   classification   MeSH
• Sensitivity and Specificity MeSH
• Nucleic Acid Amplification Techniques * methods   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• DNA, Helminth MeSH

Trematodes of the order Diplostomida are well known as serious pathogens of man, and both farm and wild animals; members of the genus Schistosoma (Schistosomatidae) are responsible for human schistosomosis (schistosomiasis) affecting more than 200 million people in tropical and subtropical countries, and infections of mammals and birds by animal schistosomes are of great veterinary importance. The order Diplostomida is also rich in species parasitizing other major taxa of vertebrates. The "Aporocotylidae" sensu lato are pathogenic in fish, "Spirorchiidae" sensu lato in reptiles. All these flukes have two-host life cycles, with asexually reproducing larvae usually in mollusks and occasionally in annelids, and adults usually live in the blood vessels of their vertebrate hosts. Pathology is frequently associated with inflammatory reactions to eggs trapped in various tissues/organs. On the other hand, the representatives of Diplostomidae and Strigeidae have three- or four-host life cycles in which vertebrates often serve not only as definitive but also as intermediate or paratenic hosts. Pathology is usually associated with migration of metacercariae and mesocercariae within the host tissues. The impact of these trematode infections on both farm and wild animals may be significant.

• Keywords
• Aporocotylidae, Blood flukes, Diplostomidae, Sanguinicolidae, Schistosoma, Schistosomatidae, Skin penetration, Spirorchiidae, Strigeidae, Trematodes,
• MeSH
• Trematode Infections * parasitology   veterinary   MeSH
• Host-Parasite Interactions MeSH
• Humans MeSH
• Schistosomatidae genetics   MeSH
• Life Cycle Stages MeSH
• Trematoda physiology   pathogenicity   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

The eggs of the blood fluke Schistosoma mansoni are the main cause of the clinical manifestations of chronic schistosomiasis. After laying, the egg "winners" attach to the endothelium of the mesenteric vein and, after a period of development, induce the growth of a small granuloma, which facilitates their passage to the intestinal lumen. Egg "losers" carried by the bloodstream to non-specific tissues also undergo full development and induce large granuloma formation, but their life ends there. Although these trapped eggs represent a dead end in the parasite life cycle, the vast majority of studies attempting to describe the biology of the S. mansoni eggs have studied these liver-trapped "losers" instead of migrating intestinal "winners". This raises the fundamental question of how these eggs differ. With robust comparative transcriptomic analysis performed on S. mansoni eggs isolated 7 weeks post infection, we show that gene expression is critically dependent on tissue localization, both in the early and late stages of development. While mitochondrial genes and venom allergen-like proteins are significantly upregulated in mature intestinal eggs, well-described egg immunomodulators IPSE/alpha-1 and omega-1, together with micro-exon genes, are predominantly expressed in liver eggs. In addition, several proteases and protease inhibitors previously implicated in egg-host interactions display clear tissue-specific gene expression patterns. These major differences in gene expression could be then reflected in the observed different ability of liver and intestinal soluble egg antigens to elicit host immune responses and in the shorter viability of miracidia hatched from liver eggs. Our comparative analysis provides a new perspective on the biology of parasite's eggs in the context of their development and tissue localization. These findings could contribute to a broader and more accurate understanding of parasite eggs interactions with the host, which have historically been often restricted to liver eggs and sometimes inaccurately generalized.

• MeSH
• Antigens, Helminth immunology   MeSH
• Liver * parasitology   immunology   metabolism   MeSH
• Mice MeSH
• Ovum metabolism   immunology   MeSH
• Helminth Proteins genetics   metabolism   immunology   MeSH
• Schistosoma mansoni * immunology   genetics   MeSH
• Schistosomiasis mansoni * immunology   parasitology   MeSH
• Intestines parasitology   immunology   MeSH
• Egg Proteins MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Antigens, Helminth MeSH
• IPSE protein, Schistosoma mansoni MeSH Browser
• Helminth Proteins MeSH
• Egg Proteins MeSH

Helminth neuroinfections represent serious medical conditions, but the diversity of the host-parasite interplay within the nervous tissue often remains poorly understood, partially due to the lack of laboratory models. Here, we investigated the neuroinvasion of the mouse spinal cord by Trichobilharzia regenti (Schistosomatidae). Active migration of T. regenti schistosomula through the mouse spinal cord induced motor deficits in hindlimbs but did not affect the general locomotion or working memory. Histological examination of the infected spinal cord revealed eosinophilic meningomyelitis with eosinophil-rich infiltrates entrapping the schistosomula. Flow cytometry and transcriptomic analysis of the spinal cord confirmed massive activation of the host immune response. Of note, we recorded striking upregulation of the major histocompatibility complex II pathway and M2-associated markers, such as arginase or chitinase-like 3. Arginase also dominated the proteins found in the microdissected tissue from the close vicinity of the migrating schistosomula, which unselectively fed on the host nervous tissue. Next, we evaluated the pathological sequelae of T. regenti neuroinvasion. While no demyelination or blood-brain barrier alterations were noticed, our transcriptomic data revealed a remarkable disruption of neurophysiological functions not yet recorded in helminth neuroinfections. We also detected DNA fragmentation at the host-schistosomulum interface, but schistosomula antigens did not affect the viability of neurons and glial cells in vitro. Collectively, altered locomotion, significant disruption of neurophysiological functions, and strong M2 polarization were the most prominent features of T. regenti neuroinvasion, making it a promising candidate for further neuroinfection research. Indeed, understanding the diversity of pathogen-related neuroinflammatory processes is a prerequisite for developing better protective measures, treatment strategies, and diagnostic tools.

• MeSH
• Arginase metabolism   MeSH
• Biomarkers metabolism   MeSH
• Chemokines metabolism   MeSH
• Eosinophils metabolism   MeSH
• Major Histocompatibility Complex MeSH
• Immunity MeSH
• Trematode Infections immunology   metabolism   pathology   MeSH
• Host-Parasite Interactions MeSH
• Spinal Cord parasitology   MeSH
• Disease Models, Animal MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Neuroglia parasitology   MeSH
• Neurons parasitology   MeSH
• Schistosomatidae immunology   MeSH
• Gene Expression Profiling MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Arginase MeSH
• Biomarkers MeSH
• Chemokines MeSH