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Sensitive diagnostics are needed to improve management and surveillance of opisthorchiasis and opisthorchiasis-associated cholangiocarcinoma (CCA) throughout East Asia. Herein we generate and screen an Opisthorchis viverrini recombinant secreted proteome to identity antibody biomarkers of liver fluke infection and CCA with sera from study participants in endemic populations and evaluate their utility as point-of-care immunochromatographic tests (PoC-ICTs). We incorporate two of the most promising antigens from the proteome array screen, P1 and P9, into PoC-ICTs to further validate their diagnostic performance. The P9-IgG4 PoC-ICT is superior amongst the single recombinant antigen tests for diagnosing fluke infection as well as fluke-induced CCA, and out-performs parasite crude extract-IgG ICTs. Here we identify two biomarkers of O. viverrini infection and infection-associated CCA that could form the basis of novel antibody serodiagnostic tests for human liver fluke infection and associated cancer. Sensitive diagnostics are needed to detect carcinogenic human liver fluke infections. Sadaow et al. here use an immunomics approach to develop field-deployable tests for diagnosing liver fluke infection and its associated liver cancer.

Before the 1990s, human fascioliasis diagnosis focused on individual patients in hospitals or health centres. Case reports were mainly from developed countries and usually concerned isolated human infection in animal endemic areas. From the mid-1990s onwards, due to the progressive description of human endemic areas and human infection reports in developing countries, but also new knowledge on clinical manifestations and pathology, new situations, hitherto neglected, entered in the global scenario. Human fascioliasis has proved to be pronouncedly more heterogeneous than previously thought, including different transmission patterns and epidemiological situations. Stool and blood techniques, the main tools for diagnosis in humans, have been improved for both patient and survey diagnosis. Present availabilities for human diagnosis are reviewed focusing on advantages and weaknesses, sample management, egg differentiation, qualitative and quantitative diagnosis, antibody and antigen detection, post-treatment monitoring and post-control surveillance. Main conclusions refer to the pronounced difficulties of diagnosing fascioliasis in humans given the different infection phases and parasite migration capacities, clinical heterogeneity, immunological complexity, different epidemiological situations and transmission patterns, the lack of a diagnostic technique covering all needs and situations, and the advisability for a combined use of different techniques, at least including a stool technique and a blood technique.

Fasciolosis is a parasitic disease caused by liver flukes of the genus Fasciola, predominantly Fasciola hepatica and Fasciola gigantica. This zoonotic disease significantly impacts both livestock and human populations, particularly in areas with extensive agriculture and poor sanitation. Ethiopia, one of Africa's leading sheep and goats producers, is highly affected by fasciolosis. However, despite its economic and public health importance, there is a lack of comprehensive and up-to-date evidence on the prevalence and risk factors of small ruminant (sheep and goats) fasciolosis. Therefore, the objective of this systematic review and meta-analysis (SRMA) is to estimate pooled prevalence and identify risk factors of fasciolosis among small ruminants in Ethiopia. This SRMA was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A comprehensive systematic review was performed across five electronic databases (Google Scholar, Embase, PubMed, Web of Science, and ScienceDirect), with all database searches and registers inquiries finalized on November 26, 2024. A random-effect model was used to determine the pooled prevalence of fasciolosis in sheep and goats. Heterogeneity was assessed, and the source of variation was analyzed using subgroup, sensitivity analysis and meta-regression. Publication bias assessment and sensitivity analysis were also performed to ensure the robustness of the review. Funnel plots and Egger's asymmetry tests were used to investigate publication bias. A total of 33 studies containing 9,578 small ruminants were included in the meta-analysis. The overall pooled prevalence of fasciolosis was 32.25% (95% CI: 25.97-38.86%) with substantially high between-study heterogeneity (inconsistency index (I2)) = 97.3%, p < 0.01). Among the variables analyzed for heterogeneity, species, publication years, season of data collection, and regions of the study were significant predictors of heterogeneity The sub-group analysis showed that the prevalence of fasciolosis among sheep and goats was 37.18% (95% CI; 31.06-43.51%) and 12.76% (95% CI; 4.06-25.19%), respectively. According to the region-based subgroup analysis, studies taken from Amhara region had the highest prevalence of fasciolosis among small ruminants (43.99% (95% CI: 31.83-56.52%)). This study highlights fasciolosis as a significant threat to Ethiopian small ruminants. Policymakers and veterinarians should prioritize evidence-based control programs in regions with high disease burden, through seasonal deworming, pasture rotation to disrupt snail ecology, and improved veterinary access in underserved areas.

The liver fluke Fasciola hepatica is a major pathogen of livestock worldwide, causing huge economic losses to agriculture, as well as 2.4 million human infections annually. Here we provide a draft genome for F. hepatica, which we find to be among the largest known pathogen genomes at 1.3 Gb. This size cannot be explained by genome duplication or expansion of a single repeat element, and remains a paradox given the burden it may impose on egg production necessary to transmit infection. Despite the potential for inbreeding by facultative self-fertilisation, substantial levels of polymorphism were found, which highlights the evolutionary potential for rapid adaptation to changes in host availability, climate change or to drug or vaccine interventions. Non-synonymous polymorphisms were elevated in genes shared with parasitic taxa, which may be particularly relevant for the ability of the parasite to adapt to a broad range of definitive mammalian and intermediate molluscan hosts. Large-scale transcriptional changes, particularly within expanded protease and tubulin families, were found as the parasite migrated from the gut, across the peritoneum and through the liver to mature in the bile ducts. We identify novel members of anti-oxidant and detoxification pathways and defined their differential expression through infection, which may explain the stage-specific efficacy of different anthelmintic drugs. The genome analysis described here provides new insights into the evolution of this important pathogen, its adaptation to the host environment and external selection pressures. This analysis also provides a platform for research into novel drugs and vaccines.

Fasciolosis, caused by the liver fluke Fasciola spp., is a significant parasitic disease of livestock and humans worldwide. Fasciola transmission and life cycle are highly dependent on climatic conditions, especially temperature and humidity. This dependency has gained significance in the context of ongoing climate change. This literature review examined evidence on the effects of temperature variability on the developmental stages of Fasciola spp. and the snail intermediate hosts. We reviewed free larval stages of Fasciola spp. development, as well as snail intermediate hosts, while investigating the climate-related factors influencing each stage. We found that Fasciola spp. egg hatching and development were inhibited below 10 °C and optimal between 20 and 30 °C, miracidia hatching time decreased with higher temperatures and cercarial shedding by snail hosts accelerated around 27 °C. Further, metacercarial viability declined at higher temperatures but was prolonged by higher humidity. Snail intermediate host growth rates peaked at 25 °C, and their susceptibility to Fasciola infection depends on temperature, underscoring its importance in transmission dynamics. Overall, the Fasciola life cycle and snail host development exhibit stage-specific temperature thresholds, indicating a complex relationship between temperature fluctuations and parasite transmission potential. This research highlights the key role of temperature and humidity on Fasciola spp. and snail development, shedding light on the potential consequences of climate change on their survival, development, and disease transmission. Data limitations, primarily from the scarcity of high-resolution climate-related experiments, should drive future research to enhance predictive models and deepen our understanding of the impact of climate change on this parasitic disease.

The study of host-parasite interactions has increased considerably in the last decades, with many studies focusing on the identification of parasite molecules (i.e. surface or excretory/secretory proteins (ESP)) as potential targets for new specific treatments and/or diagnostic tools. In parallel, in the last few years there have been significant advances in the field of extracellular vesicles research. Among these vesicles, exosomes of endocytic origin, with a characteristic size ranging from 30-100 nm, carry several atypical secreted proteins in different organisms, including parasitic protozoa. Here, we present experimental evidence for the existence of exosome-like vesicles in parasitic helminths, specifically the trematodes Echinostoma caproni and Fasciola hepatica. These microvesicles are actively released by the parasites and are taken up by host cells. Trematode extracellular vesicles contain most of the proteins previously identified as components of ESP, as confirmed by proteomic, immunogold labeling and electron microscopy studies. In addition to parasitic proteins, we also identify host proteins in these structures. The existence of extracellular vesicles explains the secretion of atypical proteins in trematodes, and the demonstration of their uptake by host cells suggests an important role for these structures in host-parasite communication, as described for other infectious agents.

Spatial heterogeneity in susceptibility and exposure to parasites is a common source of confounding variation in disease ecology studies. However, it is not known whether spatial autocorrelation acts on immunity at small scales, within wild animal populations, and whether this predicts spatial patterns in infection. Here we used a well-mixed wild population of individually recognized red deer (Cervus elaphus) inhabiting a heterogeneous landscape to investigate fine-scale spatial patterns of immunity and parasitism. We noninvasively collected 842 fecal samples from 141 females with known ranging behavior over 2 years. We quantified total and helminth-specific mucosal antibodies and counted propagules of three gastrointestinal helminth taxa. These data were analyzed with linear mixed models using the Integrated Nested Laplace Approximation, using a Stochastic Partial Differentiation Equation approach to control for and quantify spatial autocorrelation. We also investigated whether spatial patterns of immunity and parasitism changed seasonally. We discovered substantial spatial heterogeneity in general and helminth-specific antibody levels and parasitism with two helminth taxa, all of which exhibited contrasting seasonal variation in their spatial patterns. Notably, Fasciola hepatica intensity appeared to be strongly influenced by the presence of wet grazing areas, and antibody hotspots did not correlate with distributions of any parasites. Our results suggest that spatial heterogeneity may be an important factor affecting immunity and parasitism in a wide range of study systems. We discuss these findings with regards to the design of sampling regimes and public health interventions, and suggest that disease ecology studies investigate spatial heterogeneity more regularly to enhance their results, even when examining small geographic areas.

Fasciolosis, a neglected tropical disease caused by Fasciola spp., is a global parasitic infection whose prevalence is on the rise, particularly in warmer regions with favorable temperature and humidity conditions. This study examines the temporal presence of Fasciola spp. in various climatic zones across Iran, focusing on Gilan province as an endemic area. We calculated the Malone index for 5-year intervals spanning significant epidemics in 1988 and 1999 in Gilan, as well as 2005, 2010, 2015, and 2020. The results revealed a significant increase in Malone indices during August, September, and October in 1988, indicating a higher risk for Fasciola presence than in other months. These months were categorized as moderate risk. In 1999, September exhibited moderate risk, while October was classified as low risk. The risk levels remained relatively low in 2005, with the highest value in September and October. Similarly, October and September showed the highest risks in 2010, while October was identified as highly risky in 2015. In 2020, August displayed a higher risk, while September, October, and November indicated low risk. The analysis highlights the variability of risk levels across different years, with September and October consistently appearing as months with a higher probability of Fasciola presence. Conversely, the risk diminishes from January to July, which is considered a low-risk period in the region. Interestingly, a comparison with three different geographical regions in central, southern, and western Iran revealed that the Malone index was zero in all months of the year, except for two cases, suggesting unfavorable conditions for Fasciola presence in these areas. This research sheds light on the complex interplay between climatic variables and the presence of Fasciola spp., with the potential to inform the development of targeted strategies for preventing and controlling this parasite in Gilan province, a known hotspot for fasciolosis, as well as in other regions with similar climatic conditions.

Cystic Echinococcosis (CE) is a zoonotic disease caused by Echinococcus granulosus sensu lato , forming cysts in ruminants and humans with major health and economic impacts. The immune response to CE cysts is complex, with fertility linked to the host’s inflammatory reaction. This study examines γδ T cell distribution and role within the adventitial layer of non-fertile CE cysts in cattle, including cases co-infected with the trematode Fasciola hepatica (FH), a known immune response modulator. Using immunohistochemistry and double immunofluorescence, we observed γδ T cells dispersed in the adventitial layer, enriched in inflammatory zones. Co-infected cases (CE + FH+) showed a reduced γδ T cell proportion among CD3+ T cells compared to non-coinfected cases, suggesting an immunoregulatory effect of FH. Our findings align with prior studies showing γδ T cell recruitment in granulomatous diseases in ruminants but reveal that co-infection alters this response. This study provides the first detailed characterization of γδ T cells in cattle CE cysts, emphasizing their potential role in granulomatous immune responses. It highlights the need for further research into mechanisms influencing CE cyst fertility and immune modulation in helminth co-infections, advancing our understanding of host-pathogen interactions and informing disease management strategies.

Rumen fluke (Calicophoron daubneyi) has emerged as a prominent parasite of ruminants in Europe over the past decades. Epidemiological questions remain regarding this observed increase in prevalence as well as the prospect for future paramphistomosis risk. This study aimed to identify factors associated with the temporal−spatial prevalence of rumen fluke as measured by veterinary surveillance in a temperate region using zero-inflated negative binomial mixed modelling. Modelling revealed that summer rainfall, raindays and sunshine hours and mean winter temperature as significant positively associated climate variables for rumen fluke prevalence over space and time (P < 0.05). Rumen fluke prevalence was also higher in counties with higher cattle/sheep densities and was positively associated with rumen fluke case rates in the previous years (P < 0.05). Equivalent models for fasciolosis prevalence revealed no significant association with winter temperature and sunshine hours, (P > 0.05). These results confirm a strong association between rainfall and the prevalence of both fluke species in a temperate environment, likely due to the role of Galba truncatula as their intermediate snail host. It also highlights the potential added importance of winter temperature and sunshine hours in rumen fluke epidemiology when compared to liver fluke.