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Background: The liver fluke Fasciola hepatica is one of the most important endoparasites in domestic ruminants worldwide and can cause considerable economic losses. This study presents the first population genetic analysis of F. hepatica in Germany and aims at providing new insights into genetic diversity and population structure. Methods: A total of 774 liver flukes, collected from 60 cows of 17 herds and 13 cows of unknown herd origin, were subjected to comparative analysis of two mitochondrial genes (cox1 and nad1), one nuclear region (internal transcribed spacer (ITS)-1) and eight nuclear microsatellite markers. In addition, individual fluke measurements allowed comparison of morphometric differences between genotypes. Results: The nuclear ITS-1 region showed minimal variability, with 772 of 774 flukes having identical sequences, while the mitochondrial sequences revealed a high genetic diversity, with 119 distinct haplotypes, a mean haplotype diversity (Hd) of 0.81 and a mean nucleotide diversity (π) of 0.0041. Mitochondrial phylogenetic analysis identified two clusters with no clear association with the host or farm of origin. In the microsatellite analysis, all eight loci were highly polymorphic, with a mean allele frequency of 19.0 and a mean genotype frequency of 73.5 per locus. A total of 500 unique multilocus genotypes (MLGs) were found across all fluke samples, indicating that 68.5% of all genotypes were unique. A mean expected heterozygosity of 0.71 suggested a high potential for adaptability and the number of migrants (Nm = 3.5) indicated high gene flow between farms. Population structure analysis based on microsatellite data revealed that flukes from two farms differed genetically from the others. Linear mixed model results revealed that fluke length differed significantly between the two mitochondrial clusters, although it should be noted that fluke age could not be considered in the analyses. Conclusions: Fasciola hepatica in German dairy farms showed high genetic diversity and gene flow. The differences in population structure identified by mitochondrial sequences compared with microsatellite loci highlight the benefits of analysing genetic markers of different origins. This is the first study to correlate fluke morphometry measurements with genetic markers, indicating that the identified markers can influence fluke size. [ABSTRACT FROM AUTHOR]

Infection by the liver fluke, Fasciola hepatica, places a substantial burden on the global agri-food industry and poses a significant threat to human health in endemic regions. Widespread resistance to a limited arsenal of chemotherapeutics, including the frontline flukicide triclabendazole (TCBZ), renders F. hepatica control unsustainable and accentuates the need for novel therapeutic target discovery. A key facet of F. hepatica biology is a population of specialised stem cells which drive growth and development - their dysregulation is hypothesised to represent an appealing avenue for control. The exploitation of this system as a therapeutic target is impeded by a lack of understanding of the molecular mechanisms underpinning F. hepatica growth and development. Wnt signalling pathways govern a myriad of stem cell processes during embryogenesis and drive tumorigenesis in adult tissues in animals. Here, we identify five putative Wnt ligands and five Frizzled receptors in liver fluke transcriptomic datasets and find that Wnt/β-catenin signalling is most active in juveniles, the most pathogenic life stage. FISH-mediated transcript localisation revealed partitioning of the five Wnt ligands, with each displaying a distinct expression pattern, consistent with each Wnt regulating the development of different cell/tissue types. The silencing of each individual Wnt or Frizzled gene yielded significant reductions in juvenile worm growth and, in select cases, blunted the proliferation of neoblast-like cells. Notably, silencing FhCTNNB1, the key effector of the Wnt/β-catenin signal cascade led to aberrant development of the neuromuscular system which ultimately proved lethal - the first report of a lethal RNAi-induced phenotype in F. hepatica. The absence of any discernible phenotypes following the silencing of the inhibitory Wnt/β-catenin destruction complex components is consistent with low destruction complex activity in rapidly developing juvenile worms, corroborates transcriptomic expression profiles and underscores the importance of Wnt signalling as a key molecular driver of growth and development in early-stage juvenile fluke. The putative pharmacological inhibition of Wnt/β-catenin signalling using commercially available inhibitors phenocopied RNAi results and provides impetus for drug repurposing. Taken together, these data functionally and chemically validate the targeting of Wnt signalling as a novel strategy to undermine the pathogenicity of juvenile F. hepatica. Author summary: The liver fluke, Fasciola hepatica, significantly undermines the health and welfare of livestock worldwide and causes fascioliasis, a neglected tropical disease of humans. The most damaging stage of liver fluke infection is caused by the migration of juvenile worms within the liver tissue. Of all drugs approved for liver fluke treatment, just one, triclabendazole (TCBZ), is active on this pathogenic juvenile stage. TCBZ resistance is now widespread rendering liver fluke control unsustainable. This highlights the need for novel drug target identification and validation. A key aspect of juvenile worm biology is their ability to rapidly grow and develop, processes driven by a population of specialised stem cells. As such, the dysregulation of stem cells represents an attractive avenue for liver fluke control. One molecular pathway known to regulate stem cell dynamics in higher organisms is the Wnt signalling pathway. Bioinformatic searches of gene sequence datasets identified all major signalling components of both canonical and non-canonical Wnt pathways in F. hepatica. The localisation of FhWnt pathway components revealed remarkably distinct and widespread expression patterns throughout the F. hepatica body. Gene silencing of putative FhWnt pathway components revealed that those involved in the Wnt/β-catenin signal cascade are fundamental to juvenile growth and, in some cases, stem-like cell proliferation. The silencing of liver fluke β-catenin led to aberrant neuromuscular development and proved lethal to juvenile fluke. Biweekly exposures to commercially available Wnt pathway inhibitory compounds phenocopied the delayed development observed in the gene silencing experiments. These data suggest that FhWnt pathway components represent attractive targets for the development of novel flukicides or indeed, the repurposing of existing Wnt antagonists for parasite control. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is a parasitic trematode responsible for fascioliasis, a significant zoonotic disease affecting livestock worldwide, as well as humans. This study identifies peptides with potential for use in vaccines against Fasciola hepatica and validates multi-epitope constructs from those peptides in vitro. Putative protein sequences derived from the genome of F. hepatica were integrated with phase-specific transcriptomic data to prioritize highly expressed proteins. Among these, extracellular proteins were selected using DeepLoc 2.0 and strong binding affinities across diverse human and murine alleles were predicted with the IEDB MHC II tool. Peptides were further selected based on their toxicity, immunogenicity, and allergenicity. Finally, 55 high-priority candidates were obtained. To express these candidates, mRNA constructs encoding various combinations of these peptides were designed, synthesized using in vitro transcription with T7 or SP6 RNA polymerases, and transfected into cells for expression analysis. SP6 polymerase produced proper capping using CleanCapAG and was far superior in transcribing peptide constructs. Peptides fused in frame with eGFP were expressed efficiently, particularly when peptides were positioned at the 3′ terminus, opening a new field of peptide vaccines created using mRNA technology. [ABSTRACT FROM AUTHOR]

Plasmin, the final product of fibrinolysis, is a broad-spectrum serine protease that degrades extracellular matrix (ECM) components, a function exploited by multiple pathogens for dissemination purposes. The trematode Fasciola hepatica is the leading cause of fasciolosis, a major disease of livestock and an emerging zoonosis in humans. Infection success depends on the ability of F. hepatica newly excysted juveniles (FhNEJ) to penetrate the host intestinal wall, a process that remains incompletely understood. We have previously shown that FhNEJ are capable of binding plasminogen (PLG), the zymogen of plasmin, on their tegument surface, which leads to plasmin generation in the presence of host-derived PLG activators and subsequent degradation of laminin, a major component of the intestinal ECM. Here, we describe the interaction between a tegument extract of FhNEJ and the precursor of the urokinase-type PLG activator (pro-u-PA). We found that F. hepatica cathepsins B3, L3, enolase and glutathione S-transferase mediate this interaction, suggesting a multifactorial or moonlighting role for these proteins. Additionally, our results revealed that the tegument of FhNEJ contains a protease that is capable of cleaving and activating pro-u-PA into its catalytically active form, which positively impacts the capacity of the parasites to generate plasmin from the host PLG. Collectively, our findings indicate that FhNEJ interact with the host fibrinolytic system at multiple levels, reinforcing the potential of targeting this interaction as a strategy to prevent FhNEJ trans-intestinal migration and infection success. [ABSTRACT FROM AUTHOR]

The Northern Bolivian Altiplano is the fascioliasis endemic area where the highest prevalences and intensities in humans have been recorded. In this hyperendemic area of human fascioliasis, the disease is caused only by Fasciola hepatica and transmitted by Galba truncatula, the sole lymnaeid species present in the area. When analysing the link between global warning and the recently reported geographical spread of lymnaeid populations to out-border localities, a marked heterogeneous climatic change was found throughout the endemic area. The aim of the present study is to analyse the physiographical heterogeneity of the fascioliasis hyperendemic area in the Northern Bolivian Altiplano, in order to assess its repercussions in the implementation of a One Health action. We applied multivariate linear mixed models to analyse the influence of a number of physiographical features on the long-term variation of climate and of the risk of transmission. Despite its apparent physiographic homogeneity, the findings of this study revealed markedly heterogeneous climate characteristics throughout the endemic area. This irregular pattern is influenced by physiographical features such as altitude, inner hills, closeness to Lake Titicaca, and El Niño–Southern Oscillation. This is the broadest study ever performed in a human fascioliasis endemic area about the influence of physiography on climate. It highlights the importance of considering physiographical features, an aspect usually not considered in studies dealing with the influences of climate and climate change on human and animal fascioliasis. Moreover, it shows that an endemic area may climatically evolve differently in its various inner zones and emphasizes the need for continuous monitoring to assess whether control measures should be modified accordingly. Author summary: Fascioliasis is a snail-borne zoonotic disease caused by the trematodes Fasciola hepatica and Fasciola gigantica. The highest prevalences and intensities ever found in humans have been reported in the Northern Altiplano of Bolivia, in South America. In this endemic area, the disease is caused only by Fasciola hepatica and transmitted by Galba truncatula, the sole lymnaeid species present in the area. Our aim is to analyse the physiographical heterogeneity of this fascioliasis hyperendemic area, and assess its repercussions in the implementation of a One Health action. We applied multivariate linear regressions to analyse the influence of physiographical features on the long-term variation of climate and of the transmission risk. Our results revealed markedly heterogeneous climate characteristics throughout the endemic area. This irregular pattern is influenced by altitude, closeness to inner hills and Lake Titicaca, and El Niño–Southern Oscillation. These findings highlight the importance of considering physiographical features, an aspect usually not considered in studies dealing with influences of climate and climate change on human and animal fascioliasis. Moreover, it shows that an endemic area may climatically evolve differently in its various inner zones and emphasizes the need for continuous monitoring to assess whether control measures should be modified accordingly. [ABSTRACT FROM AUTHOR]

Fascioliosis is a food‐borne zoonotic helminth infection caused by flatworms belonging to the family Fasciolidae, primarily affecting ruminants. The chronic form of fascioliosis is the most prevalent and is characterized by anemia, weight loss, cirrhosis, and liver dysfunction, along with atrophy, jaundice, and bottle jaw. In humans, infection results in fever, nausea, skin rashes, and severe abdominal pain. Climate changes and human‐driven environmental alterations have contributed to an increasing incidence of fascioliosis in various regions. Fasciola species are widely distributed and have a high occurrence in tropical countries. Approximately 2.4–17 million humans are afflicted by fascioliosis in tropical and subtropical areas, with an additional 180 million facing the risk of infection. Fascioliosis poses a notable threat to ruminants; over 700 million production animals are at risk, and global annual financial losses surpass $3.2 billion. Conventional coprological methods and advanced molecular techniques are employed to diagnose fascioliosis in animals and humans. Within endemic areas, timely and accurate diagnosis is critical for successful prevention and treatment. Molecular approaches such as various PCR techniques and serological methods are extensively utilized to diagnose fascioliosis. In this review, we describe various conventional coprological and advanced DNA‐based PCR techniques along with serological methods used for the screening, monitoring, and specific diagnosis of clinical and subclinical fascioliosis in humans and animals. The information accumulated in this review will be helpful for the diagnosis of fascioliosis in the field and research laboratories. [ABSTRACT FROM AUTHOR]

Fasciolosis is a neglected tropical disease caused by helminth parasites of the genus Fasciola spp., including Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), being a major zoonotic problem of human and animal health. Its control with antihelminthics is becoming ineffective due to the increase in parasite resistance. Developing new therapeutic protocols is crucial to a deeper knowledge of the molecular bases in the host–parasite interactions. The high-throughput omics technologies have dramatically provided unprecedented insights into the complexity of the molecular host–parasite crosstalk. MicroRNAs (miRNAs) are key players as critical regulators in numerous biological processes, modifying the gene expression of cells by degradation of messenger RNA (mRNA), regulating transcription and translation functions, protein positioning, cell cycle integrity, differentiation and apoptosis. The large-scale exploration of miRNAs, including the miRNome, has offered great scientific knowledge of steps in fasciolosis, further scrutinizing the pathogenesis, the growth and development of their strains and their interaction with the host for the survival of the different parasite stages. This review compiles the updated knowledge related to miRNAs involved in fasciolosis and the generated miRNome, highlighting the importance of these key molecules in the host–parasite interactions and the pathogenesis of Fasciola spp. directing towards the development of new biotherapeutic protocols for the control of fasciolosis. [ABSTRACT FROM AUTHOR]

Bacillary hemoglobinuria (BH) is an infectious disease, mostly affecting cattle, caused by Clostridium haemolyticum (C. novyi type D), with acute hepatic necrosis and intravascular hemolysis. Cattle are typically predisposed to BH by liver injury caused by Fasciola hepatica, although cases have been reported in cattle without evidence of this parasite. Here we describe a cluster of 14 BH cases from 7 counties in north-central to central Missouri submitted to a veterinary diagnostic laboratory between December 2020 and April 2023. Postmortem examination in all cases revealed hemoglobinuria and acute hepatic necrosis with large numbers of gram-positive bacilli with terminal-to-subterminal spores. Flukes, fluke ova, and/or fluke pigment consistent with Fascioloides magna were identified in 12 of 14 cases. Sequences of the nuclear ribosomal internal transcribed spacer 1 (ITS1) from one fluke had 100% identity to F. magna. C. novyi was detected by fluorescent antibody testing of liver impression smears (11 of 12 cases) and by immunohistochemistry of liver sections (7 of 7 cases). PCR on formalin-fixed, paraffin-embedded tissues amplified the C. haemolyticum beta toxin gene in each of the 7 cases tested. To our knowledge, a confirmed cluster of BH associated with F. magna has not been reported previously in cattle. [ABSTRACT FROM AUTHOR]

Abstract Plasmin, the final product of fibrinolysis, is a broad-spectrum serine protease that degrades extracellular matrix (ECM) components, a function exploited by multiple pathogens for dissemination purposes. The trematode Fasciola hepatica is the leading cause of fasciolosis, a major disease of livestock and an emerging zoonosis in humans. Infection success depends on the ability of F. hepatica newly excysted juveniles (FhNEJ) to penetrate the host intestinal wall, a process that remains incompletely understood. We have previously shown that FhNEJ are capable of binding plasminogen (PLG), the zymogen of plasmin, on their tegument surface, which leads to plasmin generation in the presence of host-derived PLG activators and subsequent degradation of laminin, a major component of the intestinal ECM. Here, we describe the interaction between a tegument extract of FhNEJ and the precursor of the urokinase-type PLG activator (pro-u-PA). We found that F. hepatica cathepsins B3, L3, enolase and glutathione S-transferase mediate this interaction, suggesting a multifactorial or moonlighting role for these proteins. Additionally, our results revealed that the tegument of FhNEJ contains a protease that is capable of cleaving and activating pro-u-PA into its catalytically active form, which positively impacts the capacity of the parasites to generate plasmin from the host PLG. Collectively, our findings indicate that FhNEJ interact with the host fibrinolytic system at multiple levels, reinforcing the potential of targeting this interaction as a strategy to prevent FhNEJ trans-intestinal migration and infection success.

Parasitic worms (helminths) establish chronic infection within mammalian hosts by strategically regulating their host’s immune responses. Deciphering the mechanisms by which host non-coding RNAs (ncRNA) co-ordinate the activation and regulation of immune cells is essential to understanding host immunity and immune-related pathology. It is also important to comprehend how pathogens secrete specific ncRNAs to manipulate gene expression of host immune cells and influence their response to infection. To investigate the contribution of both host and helminth derived ncRNAs to the activation and/or regulation of innate immune responses during a parasite infection, we examined ncRNA expression in the peritoneal macrophages from mice infected with Fasciola hepatica. We discovered the presence of several parasitic-derived miRNAs within host macrophages at 6 hrs and 18 hrs post infection. Target prediction analysis showed that these Fasciola miRNAs regulate host genes associated with the activation of host pro-inflammatory macrophages. Concomitantly, there was a distinct shift in host ncRNA expression, which was significant at 5 days post-infection. Prediction analysis suggested that these host ncRNAs target a different cohort of host genes compared to the parasite miRNAs, although the functional outcome was predicted to be similar i.e. reduced pro-inflammatory response and the promotion of a reparative/tolerant phenotype. Taken together, these observations uncover the interplay between host and parasitic ncRNAs and reveal a complementary regulation of the immune response that allows the parasite to evade immune detection and promote tissue repair for the host. These findings will provide a new understanding of the molecular interaction between parasites and host.

Fasciola hepatica is a parasitic liver trematode that causes fasciolosis in humans and dairy animals. Traditional detection of infection is based on a microscopic examination with low sensitivity. There-fore, this study was carried out to develop an accurate and rapid method to detect F. hepatica in Al-borz province. In this cross-sectional study, 386 samples were collected from livers of cattle and sheep in three slaughterhouses of Alborz. DNA was extracted, and nested-PCR was carried out based on the F. hepatica ITS-2 sequence. DNA amplification showed first and second PCR products with expected size of 336 and 208 bp respectively, as unique for F. hepatica. The results showed no cross-reaction with the negative control. Thirty-five liver samples were positive for this parasitic infection. Totally, the prevalence of F. hepatica in the slaughterhouses of Alborz, Iran was 9.07%. This is the first study of the molecular detection of F. hepatica using nested-PCR in Alborz. It is a sensitive and rapid method that will help evaluate the prevalence rate of F. hepatica infection.

Fasciolosis is a neglected/re-emerging zoonotic disease caused by the trematode Fasciola hepatica that infects both livestock and humans worldwide, causing severe economic losses and public health problems. The study determined the seroprevalence of F. hepatica infection in buffaloes and its associations with age, sex, and climatic/environmental factors (derived from satellite data) to identify potential risk factors from two tropical climate subtypes in two Mexican states. A total of 725 serum samples were analyzed for the presence of anti-F. hepatica immunoglobulin G (IgG) antibodies using an indirect enzyme-linked immunosorbent assay (ELISA), with excretory-secretory (E/S) products as the antigen. The overall prevalence of the parasite in buffaloes between September 2022 and December 2022 was 10.62% (77 out of 725). The highest rate of F. hepatica infection was detected in Veracruz state (9.18%). In addition, the highest prevalence among the climate regions was found in the tropical rainforest (13.62%). In the two states studied, the highest positivity rate for fasciolosis was observed in young animals aged three years and below (21.52%) and in male buffaloes (20.00%). Using multivariate logistic regression analysis, we determined four factors (age, normalized difference vegetation index, land surface temperature day (LST day), and rainfall) to be significantly associated with an increased risk of parasitic infection. Rainfall was the variable most strongly associated with F. hepatica infection (OR 3.434; 95% CI: 2.181-5.406), followed by LST day (OR 2.832; 95% CI: 1.916-4.186). The results indicate that different factors are significantly associated with the prevalence of liver flukes and highlight the importance of continuous monitoring programs for parasite infections to prevent economic losses in buffalo production.

Abstract Background Climate change is driving the occurrence of several infectious diseases. Within a One Health action to complement the ongoing preventive chemotherapy initiative against human fascioliasis in the Northern Bolivian Altiplano hyperendemic area, field surveys showed a geographical expansion of its lymnaeid snail vector. To assess whether climate change underlies this spread of the infection risk area, an in-depth analysis of the long-term evolution of climatic factors relevant for Fasciola hepatica development was imperative. Methods We used monthly climatic data covering at least a 30-year period and applied two climatic risk indices, the water-budget-based system and the wet–day index, both of verified usefulness for forecasting fascioliasis transmission in this endemic area. To reveal the long-term trends of the climatic factors and forecast indices, we applied procedures of seasonal-trend decomposition based on locally weighed regression and trend analysis on the basis of linear models. To further demonstrate the changes detected, we depicted selected variables in the form of anomalies. Results This study revealed a notorious climatic change affecting most of the hyperendemic area, with a strong impact on crucial aspects of the fascioliasis transmission. Trends in maximum and mean temperatures show significant increases throughout the endemic area, while trends in minimum temperatures are more variable. Precipitation annual trends are negative in most of the localities. Trends in climatic risk indices show negative trends at lower altitudes or when farther from the eastern Andean chain. However, monthly and yearly values of climatic risk indices indicate a permanent transmission feasibility in almost every location. Conclusions Warmer temperatures have enabled lymnaeids to colonize formerly unsuitable higher altitudes, outside the endemicity area verified in the 1990s. Further, drier conditions might lead to an overexploitation of permanent water collections where lymnaeids inhabit, favoring fascioliasis transmission. Therefore, the present preventive chemotherapy by annual mass treatments is in need to widen the area of implementation. This study emphasizes the convenience for continuous monitoring of nearby zones for quick reaction and appropriate action modification. Graphical Abstract

Background: Fascioliasis is a zoonotic parasitic disease caused by Fasciola hepatica and Fasciola gigantica, which poses a serious threat to global public health and livestock farming. Fasciola gigantica secretes and excretes various components to manipulate the immune response, thereby enhancing its invasion, migration, and survival in vivo. However, the roles of specific components in immune modulation, such as asparagine endopeptidase, remain unknown. Methods: The transcriptional abundance of members of the asparagine endopeptidase family (also known as the legumain family) from F. gigantica was analyzed. Two highly transcribed asparagine endopeptidases in metacercariae, juveniles and adults were cloned, and their recombinant proteins—recombinant F. gigantica legumain (rFgLGMN-1) and (rFgLGMN-2)—were expressed in prokaryotic expression system. Their regulatory effects on buffalo peripheral blood mononuclear cells (PBMCs), including proliferation, migration, total nitric oxide (NO) production, cytokine secretion, and phagocytosis were explored in vitro. Results: Ten members of the legumain family were detected in F. gigantica, among of which FgLGMN-1 and FgLGMN-2 exhibited high transcription levels in juveniles and adults. The isolation of sequences indicated that FgLGMN-1 encodes 409 amino acids, while FgLGMN-2 encodes 403 amino acids. Both recombinant FgLGMN-1 (rFgLGMN-1) and rFgLGMN-2 were recognized by serum from buffaloes infected with F. gigantica. Both rFgLGMN-1 and rFgLGMN-2 inhibited the proliferation of PBMCs, and rFgLGMN-1 also inhibited the migration of PBMCs. While rFgLGMN-1 increased the production of total NO, rFgLGMN-2 decreased NO production. Both rFgLGMN-1 and rFgLGMN-2 increased the transcription of the cytokines interleukin-10 and transforming growth factor β. The effect of rFgLGMN-1 and rFgLGMN-2 on the phagocytosis of PBMCs varied depending on their concentrations. Conclusions: rFgLGMN-1 and rFgLGMN-2 modulate several cellular and immunological functions of PBMCs, and exhibited distinct regulatory effects on these in vitro, which indicated that they may play roles in immune modulation and facilitate fluke development. However, due to uncertainties associated with in vitro experiments, further studies are necessary to elucidate the precise functions of these legumains. [ABSTRACT FROM AUTHOR]

As animals age, they exhibit a suite of phenotypic changes, often including reductions in movement and social behaviour ('behavioural ageing'). By altering an individual's exposure to parasites, behavioural ageing may influence infection status trajectories over the lifespan. However, these processes could be confounded by age-related changes in other phenotypic traits, or by selective disappearance of certain individuals owing to parasite-induced mortality. Here, we uncover contrasting age-related patterns of infection across three helminth parasites in wild adult female red deer (Cervus elaphus). Counts of strongyle nematodes (order: Strongylida) increased with age, while counts of liver fluke (Fasciola hepatica) and tissue worm (Elaphostrongylus cervi) decreased, and lungworm (Dictyocaulus) counts did not change. These relationships could not be explained by socio-spatial behaviours, spatial structuring, or selective disappearance, suggesting behavioural ageing is unlikely to be responsible for driving age trends. Instead, social connectedness and strongyle infection were positively correlated, such that direct age–infection trends were directly contrasted with the effects implied by previously documented behavioural ageing. This suggests that behavioural ageing may reduce parasite exposure, potentially countering other age-related changes. These findings demonstrate that different parasites can show contrasting age trajectories depending on diverse intrinsic and extrinsic factors, and that behaviour's role in these processes is likely to be complex and multidirectional. This article is part of the discussion meeting issue 'Understanding age and society using natural populations'. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is a trematode parasite of significant veterinary and public health importance, causing economic losses in livestock due to liver damage, weight loss and reduced milk production. Although triclabendazole (TCZ) is available for treatment, it does not prevent the disease or reinfection. Infected animals exhibit strong immunoregulation, increasing susceptibility to secondary infections and altering vaccine‐induced antibody responses. This study investigates the humoral immune response in cattle infected with F. hepatica at different stages of infection and evaluates the effect of TCZ treatment on this response. It also examines how fasciolosis affects the antibody response induced by bacterial vaccines during early and chronic infection stages. Experimental infections in steers were conducted, with faecal and plasma samples collected at various intervals. The results showed a decrease in parasite‐specific antibody avidity during infection. However, F. hepatica infection did not substantially modify antibody response to bacterial vaccines. This study underscores the need for further research on the impact of fasciolosis and its treatment on livestock vaccination efficacy. [ABSTRACT FROM AUTHOR]

Simple Summary: Fasciolosis is one of the parasitic diseases that exerts the greatest impact on sheep productive systems worldwide. Affected sheep generally show a decrease in their reproductive capacity, weight gain, meat and milk production, and wool quality. In the pathogenesis of Fasciola hepatica, the penetration and migration of parasitic stages through the liver provoke intense inflammatory immune responses and tissue damage. Following strong activation signals, polymorphonuclear neutrophils (PMNs) reportedly release chromatin and granular proteins into the extracellular space, forming DNA traps called neutrophil extracellular traps (NETs). It was recently demonstrated that F. hepatica induces NET formation in vivo in the liver parenchyma from naturally infected sheep. Thus, in this paper we investigated F. hepatica-mediated NET-derived cytotoxic effects in exposed hepatic cells in vitro as well as F. hepatica-triggered ovine NETs under hypoxic conditions (5% O2). Our results confirm the ability of ovine PMNs to form NETs in low-oxygen environments after F. hepatica antigen (FhAg) stimulation, as seen in many pathological and pro-inflammatory states, and their cytotoxic effects on exposed hepatic cells. In addition, we demonstrate that this defence mechanism is NADPH oxidase (NOX)-dependent and therefore a tightly regulated molecular process. Overall, we hypothesize that NET formation plays a role in the pathogenesis of fasciolosis, and NETs contribute to liver tissue damage if released in an uncontrolled manner. Fasciola hepatica is a parasitic trematode that causes fasciolosis in sheep, provoking a decrease in their reproductive capacity, weight gain, meat and milk production, and wool quality. In the pathogenesis of F. hepatica, the penetration and migration of parasitic stages through the liver provoke intense inflammatory immune responses and tissue damage. The aim of this study was to investigate the cytotoxic effects of Fascila hepatica-induced ovine NETs in exposed hepatocytes in vitro, and to analyze whether F. hepatica antigens (FhAg) trigger the release of ovine NETs under hypoxic conditions as well as the roles of matrix metalloproteinase-9 (MMP-9) and CD11b in this cellular process in vitro. Here, isolated ovine PMNs were co-cultured with FhAg under hypoxia (5% O2) and NETs were visualized via immunofluorescence analyses, confirming their classical characteristics. The quantification of NETs in response to FhAg in hypoxic conditions significantly enhanced the formation of anchored and cell-free NETs (p < 0.01), and NADPH oxidase (NOX) inhibitor diphenylene iodonium (DPI) significantly reduced their production (p < 0.05). Furthermore, the cytotoxic effect of NETs on hepatic cells was determined by using a live/dead-staining with Sytox Orange, thereby demonstrating that FhAg-induced NETs are cytotoxic for hepatic cells (p = 0.001). We additionally analyzed PMN supernatants to determine the enzymatic activity of MMP-9, observing that FhAg exposure enhances MMP-9 release in ovine PMNs (p < 0.05) but not in bovine PMNs. Interestingly, by using flow cytometric analysis, we determined that the exposure of PMNs to FhAg does not increase the CD11b surface expression of ovine PMNs. This could be an effect of the activation of other surface receptors or transcription factors involved in F. hepatica-induced NETosis. Consequently, we hypothesize that F. hepatica-induced NETs play a role in the pathogenesis of fasciolosis, contributing to liver tissue damage if released in an uncontrolled manner. [ABSTRACT FROM AUTHOR]

Background and Aim: Fasciola spp. are important trematodes of public health concern in various animals, including camels. This study aimed to determine the occurrence of liver flukes in camels, to determine the molecular confirmation of Fasciola, and to perform phylogenetic analysis of study isolates to identify the species of Fasciola. Materials and Methods: In total, 107 slaughtered camels were inspected to collect liver flukes that were examined molecularly using polymerase chain reaction (PCR) to confirm Fasciola species. Then, the study isolates were sequenced, submitted to the National Center for Biotechnology Information (NCBI) database, and analyzed phylogenetically to identify the species of each study isolate. Results: Liver flukes were detected in 17.67% of the camels. Regarding the migratory stages of the collected worms, juvenile worms (73.91%) were significantly more prevalent than adult worms (26.09%). Regarding the risk factors, a significantly greater occurrence rate and risk of infection was detected in Al-Najaf compared with Al-Muthanna, as well as in younger camels (1-4 years) compared with older camels. Although the occurrence rate of liver flukes was insignificantly different between females (9.38%) and males (6.98%), females appeared to be at a significantly higher risk of infection than males. Molecularly, 33.33% of the worm samples were positive for species in the Fasciola genus. Phylogenetic analysis of all positive PCR products (total no = 19) confirmed that 63.16% of the local Fasciola spp. isolates were related to the NCBI-Basic Local Alignment Search Tool (NCBI-BLAST) Saudi Arabian Fasciola hepatica isolate at an identity range of 95.94%-99%; while 36.84% of the local Fasciola spp. isolates were related to the NCBI-BLAST Iranian F. gigantica isolate at an identity range of 97.73%-99%. Conclusion: This study found a 17.67% occurrence of Fasciola spp. in camels, with juvenile worms being more common than adult worms. Molecular analysis revealed that 63.16% of the isolates were related to F. hepatica from Saudi Arabia, while 36.84% matched F. gigantica from Iran. Younger camels and those from Al-Najaf were at higher risk, highlighting the need for targeted control measures. [ABSTRACT FROM AUTHOR]

Background: The trematode parasite Fasciola hepatica (liver fluke) can infect livestock, wild mammals, and humans, generating serious economic losses worldwide. Aquatic or amphibious snails of the Lymnaeidae family are the intermediate host of this parasite. Both snail population dynamics and parasite development are closely associated with temperature, although most field studies have recorded air temperature rather than water temperature. Our aim was to statistically model the population dynamics of lymnaeid snails and their infection by F. hepatica under natural environmental conditions in Northwest Andean Patagonia. Methods: For two years, we sampled snails monthly in four bodies of water, while registering water and air temperature hourly, and assessing F. hepatica infection in snails. Hierarchical Bayesian modeling allowed us to estimate the functional relationship between water temperature and population growth, the probability of detecting snails, and infection by F. hepatica. Results: A total of 1,411 Galba viatrix snails were collected, identified, and analyzed for F. hepatica infection. All sites showed seasonal variation in the number of snails collected and in water temperature as well as sharp variations in snail counts between surveys adjacent in time. The hierarchical model revealed that water temperature acts, at least, at two different time scales: water temperature at the time of sampling determines snail detection probability, whereas the average water temperature between sampling dates affects lymnaeid population growth. We found maximum F. hepatica prevalences in snails of 40% (2/5 and 4/10), followed by 33% (65/197). These are the highest prevalences recorded in G. viatrix populations in Argentina to date. Our modeling evidenced that the positive effects of water temperature on infection probability increases with snail size and prevalence on the previous survey, while previous prevalence strongly enhances the effects of snail size. Conclusions: Our results underscore the high temporal and spatial variability in the population of snails and the prevalence of F. hepatica, as well as the major impact temperature has on detecting snails. Our models provide quantifications of the effects of water temperature on the population growth of G. viatrix, its detection, and infection under natural field conditions. These are crucial steps towards generating mechanistic models of F. hepatica transmission that would facilitate the design and simulation of potential interventions based on treatments and on environmental and livestock management, taking into account the specific characteristics of each region. [ABSTRACT FROM AUTHOR]

Purpose: The objective of the present work was to evaluate the effect of an ethyl acetate extract of Artemisia ludoviciana on the viability of adult Fasciola hepatica parasites and eggs. Methods: The collection of plant material was performed as described in previous reports. The dried material was macerated with ethyl acetate. Ovicidal assays were performed at concentrations of 100, 200, 300, 400 and 500 mg/L A. ludoviciana extract. Bioassays of fasciolicidal efficacy in adult specimens of F. hepatica were performed at extract concentrations of 125, 250, 375 and 500 mg/L. The effects of triclabendazole, a reference drug, and artemisinin were also evaluated. Results: The ovicidal effectiveness of the extracts obtained after 16 h of incubation at concentrations of 100, 200, 300, 400 and 500 mg/L was 48%, 52%, 87%, 89% and 92%, respectively (p < 0.05), and the fasciolicidal efficiencies during the first 24 h post-treatment ranged from 82 to 100% (p < 0.05). In both cases, scanning electron microscopy revealed damage to the shells of the eggs treated with the extract, compromising their stability. In adult fasciolae, alterations to the integument that resulted in its erosion and detachment were observed. Histopathological studies of the affected specimens revealed damage to the tegumentary and subtegumentary cells and alterations in the ovaries, testicles and intestine. This damage was more severe after treatment with the extract than after treatment with the other compounds. Methods: Extract of A. ludoviciana causes damage to the tegument, intestine, ovaries, testes and eggs of F. hepatica. Highlights: •Evaluate the effect of an ethyl acetate extract of Artemisia ludoviciana on the viability of adult Fasciola hepatica parasites and eggs. •The ovicidal effectiveness of the extracts obtained after 16 h of incubation at concentrations of 100, 200, 300, 400 and 500 mg/L was 48%, 52%, 87%, 89% and 92%, respectively (p<0.05), and the fasciolicidal efficiencies during the first 24 h post-treatment ranged from 82% to 100% (p<0.05). •Scanning electron microscopy revealed damage to the shells of the eggs treated with the extract, compromising their stability. •In parasites exposed to extract exhibited partial erosion of the integument in different regions, damage to the tegument that affected the integrity of the spines was seen. In some cases, severe damage to the muscles was observed. •Histological evaluation observe areas of advanced deterioration and regions. Alterations at this level allow substances access to deep organs, which can then be damaged. In this study, extract affected virtually the entire interior of the organism, including the digestive tract, the gonads, the internal parenchyma, and the subepidermal cells. [ABSTRACT FROM AUTHOR]

Purpose: Fascioliasis is a zoonotic parasitic disease spread by genus Fasciola that causes considerable health and economic losses to both humans and livestock. Pakistan is an endemic region for fascioliasis and only a few molecular studies have been conducted to describe the etiology of disease in the country. Methods: Current study involved the molecular characterization of adult fluke specimens (n = 31) through ITS-1 region of rDNA collected from four host species (sheep, goats, cattle and buffaloes) of district Sargodha. Moreover, coprology based epidemiology (n = 28620) and association of different risk factors with disease prevalence was also determined. Additionally, an overview of prevalence of fascioliasis in Pakistan, including molecular studies in the four provinces and livestock species, was also carried out. Results: The phylogenetic analysis revealed that Fasciola gigantica was the main etiological agent infecting the livestock species of this region. The coprological survey revealed an overall prevalence of 30.18% among the livestock species (sheep = 32.31%, goats = 30.67%, cattle = 31.01%, buffaloes = 25.23%). It was inferred that females were slightly more susceptible to infection (32.59%) and that the older age group had higher infection rate (34.01%) in all the host species. Conclusion: The current study suggests that fascioliasis is a disease of primary concern indicating its status as one of the prevalent diseases affecting livestock in this region. It was also concluded that both Fasciola gigantica and Fasciola hepatica existed among the infected animals in addition to presence of hybrid/intermediate form in Pakistan with F. gigantica as dominant species in Punjab province. [ABSTRACT FROM AUTHOR]

Fasciolosis is a parasitosis of great importance for livestock, as well as for public health, as it is considered by the WHO as a neglected disease. Disease control is complex and reinfections make the use of therapeutic products an unsustainable method from an economic, environmental and health point of view. The aim of this study was to evaluate a new vaccine formulation for dairy cattle, containing soluble Fasciola hepatica antigens associated with Montanide 763 AVG and saponin adjuvants (FhSAMS). The vaccine was tested with two protocols, a single dose and a booster dose 6 months after the first dose. The FhSAMS vaccine proved to be safe, with no side effects. Furthermore, it was able to generate a more robust humoral immune response when a six‐month booster dose was used, in addition to stimulating greater production of IFN‐ʏ, indicating a Th1 profile immune stimulus. [ABSTRACT FROM AUTHOR]

Background: Liver flukes from the genus Fasciola are the causative agents for human and livestock fascioliasis. Accurate identification of Fasciola spp. is essential to understanding the epidemiology of fascioliasis. This study aimed to determine the morphometric and molecular characterization of Fasciola spp. in livestock from Northwestern provinces of Iran. Methods: Five hundred adult Fasciola flukes were obtained from different definitive hosts (cattle, sheep, goats, and buffaloes) in four local abattoirs in the northwestern provinces of Iran (West‐Azerbaijan, East‐Azerbaijan, Ardabil, and Zanjan) from September 2021 to August 2022. All samples were examined by morphometric criteria; then, 49 samples were identified using PCR‐RFLP based on ITS1 region and 23 sequence of isolates analyzed by cox1 marker. PCR‐RFLP methods compared morphometric results, and cox1 gene sequences were used to confirm PCR‐RFLP results and phylogenetic analysis. Results: The differences between the body length, body width, cephalic cone length, cephalic cone width, body area, and distance between the ventral sucker and posterior end of the body in two species were significant (p < 0.05). Based on the morphometric criteria, 139 samples (27.8%) were identified as Fasciola gigantica and 361 (72.2%) as Fasciola hepatica. Similarly, PCR‐RFLP analysis of ITS1 region confirmed morphometric results. No hybrid forms of Fasciola were detected. Partial sequences of cox1 showed 13 variable sites with eight haplotypes in F. hepatica and 12 variable sites with five haplotypes in F. gigantica. Conclusion: Based on the results of this study, the PCR‐RFLP method can be used to confirm the morphological method of Fasciola species, but it is insufficient to study their genetic diversity. Also, sequences of cox1 results of the present study showed that F. hepatica and F. gigantica species in the Northern provinces of Iran have different genetic structures and haplotypes. [ABSTRACT FROM AUTHOR]

Fasciolosis is a zoonotic infection and is considered a developing deserted tropical illness threatening ruminant productivity and causing financial losses. Herein, we applied immunoinformatics and biophysics studies to develop an epitopes vaccine against Fasciola hepatica using glutathione transferase and Cathepsin L-like proteinase as possible vaccine candidates. Using the selected proteins, B- and Tcell epitopes were predicted. After epitopes prediction, the epitopes were clarified over immunoinformatics screening, and only five epitopes, EFGRWQQEKCTIDLD, RRNIWEKNVKHIQEH, FKAKYLTEMSRASDI, TDMTFEEFKAKYLTE, and YTAVEGQCR were selected for vaccine construction; selected epitopes were linked with the help of a GPGPG linker and attached with an adjuvant through another linker, EAAAK linker. Cholera toxin B subunit was used as an adjuvant. The ExPASy ProtParam tool server predicted 234 amino acids, 25.86257 kDa molecular weight, 8.54 theoretical pI, 36.86 instability index, and -0.424 grand average of hydropathicity. Molecular docking analysis predicted that the vaccine could activate the immune system against F. hepatica. We calculated negative binding energy values. A biophysics study, likely molecular docking molecular dynamic simulation, further validated the docking results. In molecular dynamic simulation analysis, the top hit docked compounds with the lowest binding energy values were subjected to MD simulation; the simulation analysis showed that the vaccine and immune cell receptors are stable and can activate the immune system. MMGBSA of -146.27 net energy (kcal/mol) was calculated for the vaccine-TLR2 complex, while vaccine-TLR4 of -148.11 net energy (kcal/mol) was estimated. Furthermore, the C-ImmSim bioinformatics tool predicted that the vaccine construct can activate the immune system against F. hepatica, eradicate the infection caused by F. hepatica, and reduce financial losses that need to be spent while protecting against infections of F. hepatica. The computational immune simulation unveils that the vaccine model can activate the immune system against F. hepatica; hence, the experimental scientist can validate the finding accomplished through computational approaches. [ABSTRACT FROM AUTHOR]

Background: Climate change is driving the occurrence of several infectious diseases. Within a One Health action to complement the ongoing preventive chemotherapy initiative against human fascioliasis in the Northern Bolivian Altiplano hyperendemic area, field surveys showed a geographical expansion of its lymnaeid snail vector. To assess whether climate change underlies this spread of the infection risk area, an in-depth analysis of the long-term evolution of climatic factors relevant for Fasciola hepatica development was imperative. Methods: We used monthly climatic data covering at least a 30-year period and applied two climatic risk indices, the water-budget-based system and the wet–day index, both of verified usefulness for forecasting fascioliasis transmission in this endemic area. To reveal the long-term trends of the climatic factors and forecast indices, we applied procedures of seasonal-trend decomposition based on locally weighed regression and trend analysis on the basis of linear models. To further demonstrate the changes detected, we depicted selected variables in the form of anomalies. Results: This study revealed a notorious climatic change affecting most of the hyperendemic area, with a strong impact on crucial aspects of the fascioliasis transmission. Trends in maximum and mean temperatures show significant increases throughout the endemic area, while trends in minimum temperatures are more variable. Precipitation annual trends are negative in most of the localities. Trends in climatic risk indices show negative trends at lower altitudes or when farther from the eastern Andean chain. However, monthly and yearly values of climatic risk indices indicate a permanent transmission feasibility in almost every location. Conclusions: Warmer temperatures have enabled lymnaeids to colonize formerly unsuitable higher altitudes, outside the endemicity area verified in the 1990s. Further, drier conditions might lead to an overexploitation of permanent water collections where lymnaeids inhabit, favoring fascioliasis transmission. Therefore, the present preventive chemotherapy by annual mass treatments is in need to widen the area of implementation. This study emphasizes the convenience for continuous monitoring of nearby zones for quick reaction and appropriate action modification. [ABSTRACT FROM AUTHOR]

The spatial organization of gene expression dictates tissue functions in multicellular parasites. Here, we present the spatial transcriptome of a parasitic flatworm, the common liver fluke Fasciola hepatica. We identify gene expression profiles and marker genes for eight distinct tissues and validate the latter by in situ hybridization. To demonstrate the power of our spatial atlas, we focus on genes with substantial medical importance, including vaccine candidates (Ly6 proteins) and drug resistance genes (glutathione S-transferases, ABC transporters). Several of these genes exhibit unique expression patterns, indicating tissue-specific biological functions. Notably, the prioritization of tegumental protein kinases identifies a PKCβ, for which small-molecule targeting causes parasite death. Our comprehensive gene expression map provides unprecedented molecular insights into the organ systems of this complex parasitic organism, serving as a valuable tool for both basic and applied research. Fasciola hepatica is a complex multicellular pathogen and the causative agent of fascioliasis, a zoonotic disease that compromises liver function. Using spatial transcriptomics, the authors uncover the tissue-specific gene expression of F. hepatica and identify genes with expression in the surface layer for drug-target discovery. [ABSTRACT FROM AUTHOR]

Introduction: Infections with Fasciola hepatica and Ostertagia ostertagi impinge upon dairy cow health and welfare and represent a major economic factor in livestock industry. Control measures largely rely on the use of anthelminthic drugs. However, reports of anthelmintic resistance necessitate sustainable approaches. Farmer characteristics such as attitude and personality are crucial for the implementation of control strategies and on-farm practices. Methods: In the present study, the HEXACO (Honesty-Humility, Emotionality, eXtraversion, Agreeableness, Conscientiousness, Openness to experience) model of personality structure, which conceptualizes human personality, was used to evaluate the relationship of farmer aspects with on-farm bulk tank milk seropositivity for F. hepatica and O. ostertagi. Moreover, information on farm structure, housing, management, and farmers' attitude was collected in a face-to-face interview. Farm-level seropositivity for F. hepatica and O. ostertagi was predicted via elastic net regression. Results: Out of 193 farms housing 8,774 cows in the German Federal State of Bavaria, 47 farms (24.4%) were seropositive for F. hepatica, 77 farms (39.9%) for O. ostertagi, and 42 farms (21.8%) for both endoparasites. The model for F. hepatica seropositivity selected the covariates pasture access, O. ostertagi seropositivity, higher farmer conscientiousness, and organic farming as relevant predictors. Seropositivity for O. ostertagi was predicted by F. hepatica seropositivity, pasture access, organic farming, and farmers being neutral regarding their satisfaction with animal health on their farm. Higher values for the HEXACO factors extraversion and emotionality were inversely associated with O. ostertagi seropositivity. Discussion: The present work emphasizes the importance of farmer traits in regard to animal health and parasite occurrence. For the effective acceptance and implementation of sustainable control strategies for livestock helminth infections, it is crucial to consider these aspects to holistically address the challenges of managing parasitic diseases. Moreover, tailored communication strategies can be developed incorporating the understanding of individual stockman characteristics and subsequently ensuring encouragement of stakeholders. [ABSTRACT FROM AUTHOR]

[Display omitted] • Fasciola hepatica extracellular vesicles (Hp EVs) induce extracellular matrix secretion. • Hp EVs reduce proliferation of hepatic stellate cells (HSCs) • Dicrocoelium dendriticum EVs (Dd EVs) trigger a global inflammatory response in HSCs and hepatocytes. • Dd EVs have a more potent anti-proliferative effect than Fh EVs. • Fh EVs are taken up faster and in higher amounts than Dd EVs by HSCs and hepatocytes. Fasciola hepatica and Dicrocoelium dendriticum are parasitic trematodes residing in the bile ducts of mammalian hosts, causing, in some cases, impairment of liver function and hepatic fibrosis. Previous studies have shown that extracellular vesicles released by F. hepatica (Fh EVs) and D. dendriticum (Dd EVs) induce a distinct phenotype in human macrophages, but there is limited information on the effect of parasitic EVs on liver cells, which interact directly with the worms in natural infections. In this study, we isolated Fh EVs and Dd EVs by size exclusion chromatography and labeled them with a lipophilic fluorescent dye to analyze their uptake by human hepatic stellate cells (HSC) and hepatocytes, important cell types in liver pathology, using synthetic liposomes as internal labeling and uptake control. We analyzed EV uptake and the proteome profiles after the treatment with EVs for both cell types. Our results reveal that EVs establish unique and specific interactions with stellate cells and hepatocytes, suggesting a different role of EVs derived from each parasite, depending on the migration route to reach their final niche. Fh EVs have a cytostatic effect on HSCs, but induce the extracellular matrix secretion and elicit anti-inflammatory responses in hepatocytes. Dd EVs have a more potent anti-proliferative effect than Fh EVs and trigger a global inflammatory response, increasing the levels of NF-κB and other inflammatory mediators in both cell types. These interactions may have a major influence on the progression of the disease, serving to generate conditions that may favor the establishment of the helminths in the host. [ABSTRACT FROM AUTHOR]

This article discusses Fasciola hepatica infection, a zoonotic parasite that lives in the liver bile ducts. A 31-year-old female patient was diagnosed with symptoms such as nausea, increased liver enzymes, and right upper quadrant pain for about a year. The parasite was detected in the common bile duct by Endoscopic Ultrasound (EUS) and removed by Endoscopic Retrograde Cholangio Pancreatography (ERCP). Treatment was performed with 10 mg/kg triclabendazole. Eosinophilia, abdominal pain, and dietary history are important clues in the diagnosis of infection. Imaging methods, especially EUS, play a critical role in diagnosis. With this method, parasites can be seen as mobile hyperechogenic structures. If untreated, parasites can survive in their hosts for many years, therefore early diagnosis and treatment are important in preventing complications. It is recommended to monitor the eosinophil levels and serological test results of patients after treatment. As a result, EUS is a very valuable diagnostic tool in suspected cases. [ABSTRACT FROM AUTHOR]

Fasciolosis (liver fluke disease) is a zoonosis of significant public health relevance, caused by parasites of the genus Fasciola, specifically of the species Fasciola hepatica, a helminth of the class Trematoda, which exhibits a lanceolate morphology and is involved in the zoonotic cycle through contamination of bodies of water and aquatic vegetation, including watercress, by means of metacercariae, its infective form. Their biological cycle includes a migration phase from the host's intestine to the liver, where they mature and settle in the bile ducts, causing pathological changes such as direct hepatic lesions, bile duct obstruction and a systemic inflammatory response. Clinically, these events manifest through predominantly hepatic and gastrointestinal symptoms secondary to hepatic involvement, which can escalate to severe complications such as cirrhosis and hepatic abscesses in the absence of early diagnosis and adequate treatment. We present the case of a 54-year-old female patient who was admitted to the emergency room with a condition characterized by headache, vomiting, cough, palpitations and dyspnea on exertion. The initial diagnostic evaluation, guided by the ultrasound presentation of choledocholithiasis, led to an endoscopic retrograde cholangiopancreatography (ERCP) that revealed the presence of a single papilla. This finding, along with the described symptoms, facilitated the diagnosis of hepatic fasciolosis caused by Fasciola hepatica, which was later removed. Likewise, it underscores the importance of including this parasitosis as a differential diagnosis for hepatic and gastrointestinal diseases, particularly in compatible epidemiological contexts or a history of exposure to contaminated freshwater sources. [ABSTRACT FROM AUTHOR]

Fascioliasis is an often-neglected zoonotic disease caused by liver fluke Fasciola hepatica and Fasciolagigantica[1]. The life cycle of this parasite starts when eggs in mammalian stool are deposited in tepid water (22-26°C) and hatch in within two weeks, miracidia appear and develop. These miracidia invade many species of freshwater snails, in which they further develop to sporocyst and redia for 4-7 weeks. They leave as free-swimming single tailed cercaria that subsequently attach to watercress, water lettuce, mint, parsley, or khat [2, 3]. They encyst within few hours and wait to be eaten by the definite host, humans [4]. The metacercariae exist in the small intestine is releasing the young parasites, which rapidly penetrate the intestinal wall and enter the peritoneal cavity. The immature flukes penetrate the capsule of Glisson after 48h and enter the liver then migrate throughout the hepatic parenchyma till they reach the biliary system where they become adults within 3 to 4 months from the initial infection and lay eggs [4]. World health organization now recognizes human Fascioliasis as a significant public health problem and a neglected tropical disease with a great impact on human development [4]. The burden of infection is unclear, and studies have shown the geographic expansion of fascioliasis in human and livestock likely related to climate change[5]. This study aims to highlight the sociodemographic characteristics of human fascioliasis in a newly emerged endemic area at north of Iraq. [ABSTRACT FROM AUTHOR]

Three studies were appraised. This included two cross-sectional diagnostic accuracy studies and one case control diagnostic accuracy study. Moderate. The first study reported the findings from 619 tested cattle over 3 sample periods comparing the sensitivity and specificity of the different tests. The sensitivity of the faecal egg sedimentation test varied greatly between the sample periods from 0.81 (95% beta coefficient (BCI) 0.72-0.90) to 0.58 (95% BCI 0.43-0.72) with the coproantigen ELISAs sensitivity remaining consistent at 0.77 (95% BCI 0.64-0.88) throughout. The second study reported the findings of 200 tested cattle over 2 sampling periods comparing the sensitivity and specificity of the different tests. The mean sensitivity of the coproantigen ELISA was significantly higher than the 4 g/10 g preparations of the faecal egg sedimentation tests at 94% (95% CI 87%-98%) (P < 0.001). The third study reported the findings of coproantigen ELISA testing on 250 bovine faecal samples with 94 confirmed positive for liver fluke via faecal sedimentation testing. The sensitivity of the coproantigen ELISA was 80% and the specificity was 100% compared with 70% and 80% respectively for the faecal egg sedimentation test. All three studies demonstrated either an increased or equivalent sensitivity of the coproantigen ELISA to the faecal sedimentation test, but only one study reported a statistically significant increase in test sensitivity. Whilst all three studies were diagnostic accuracy validity studies, the systematic sampling strategy of one study was superior to the convenience sampling method of one of the other studies and to the case control method of the other. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is a liver fluke that resides in the bile ducts of various mammals. The parasitosis leads to economic losses in animal production estimated at USD 3.2 billion annually. It is also considered a zoonosis of great significance and a problem for public health affecting 2.4 million people worldwide. Nevertheless, besides the negative aspects of infestation, the antigens released by the fluke, F. hepatica Excretory-Secretory Products (Fh-ES) contain several immunomodulatory molecules that may be beneficial during the course of type I diabetes, multiple sclerosis, ulcerative colitis, or septic shock. This phenomenon is based on the natural abilities of adult F. hepatica to suppress proinflammatory responses. To underline the molecular basis of these mechanisms and determine the role of microRNA (miRNA) in the process, lipopolysaccharide (LPS)-activated THP-1 macrophages were stimulated with Fh-ES, followed by miRNA microarray analyses. Surprisingly, no results indicating changes in the miRNA expression profile were noted (p < 0.05). We discuss potential reasons for these results, which may be due to insufficient sensitivity to detect slight changes in miRNA expression or the possibility that these changes are not regulated by miRNA. Despite the negative data, this work may contribute to the future planning of experiments by other researchers. [ABSTRACT FROM AUTHOR]

ABSTRACT Background Fascioliasis represents one of the most significant parasitic and foodborne zoonotic diseases in the world. Resistance to currently deployed human and veterinary flukicides is a growing health problem. Zinc oxide nanoparticles (ZnO‐NPs) have developed enormous importance in nanomedicine. The current study aims to evaluate the flukicide activity of ZnO‐NPs on the adult stages of Fasciola hepatica, in vitro assay. Methods Mature flukes of F. hepatica were collected from the biliary tracts of cattle liver from the main slaughterhouses in Aswan Governorate, Egypt. The co‐precipitation process was utilized to create ZnO‐NPs. The adult active flukes of F. hepatica (n = 6) were transferred in Petri dishes (triplicates) containing different concentrations of ZnO‐NPs (0.05, 0.10 and 0.12 mg/mL), with simultaneous maintenance of control in only PBS. The treated and control groups of flukes were maintained inside an incubator at 37°C ± 1°C. Mortality and paralysis of the flukes were observed. The adult flukes were picked up for scanning electron microscopy (SEM) examination from the control medium, and those exposed to the highest effective concentration of ZnO‐NPs. ZnO‐NPs showed dose‐dependent flukicide efficacy against the adult flukes. Results At 0.12 mg/mL concentration of ZnO‐NPs, the mortality of flukes was attained early in 4.05 ± 0.033 h. The photomicrographs obtained by SEM of the flukes revealed that ZnO‐NPs (0.12 mg/mL) had a potent effect on the integument surface and genital system of F. hepatica. Most of the tegument was damaged with complete erosion and loss of the spines and papillae. Furthermore, a severe rupture in the genital system causes the uterus to burst and releases eggs through the genital pore. Conclusions Hence, it could be concluded that ZnO‐NPs performed effective anthelmintic activity against the adult stages of F. hepatica, in vitro. The results of the present study recommend the use of ZnO‐NPs as new preparations with anthelmintic activity.

Abstract Background Infections with (tricho-)strongyles, Dictyocaulus viviparus or Fasciola hepatica have been shown to reduce milk production in dairy cows. However, the current published studies focused on one single helminth infection by neglecting helminth co-infections and their possible (additive) effects on host performance. Hence, for the first time, we investigated differences in the impact of patent helminth co-infections versus mono-infections on milk production parameters in individual cows. Methods A total of 1583 dairy cows from 27 herds were included in this study. Faecal samples were examined in 2015 and 2021/2022 to determine the number of eggs/larvae per gram faeces for (tricho-)strongyles, D. viviparus, F. hepatica and rumen flukes. The cows were classified as non-infected, mono-infected and co-infected. Linear mixed models were applied to analyse the association between infection status (non-infected vs. mono-infected vs. co-infected) with milk yield, milk protein and milk fat content by including potential confounders. Results Infections with (tricho-)strongyles, D. viviparus, F. hepatica and rumen flukes were detected in 100%, 28.6%, 50.0% and 21.4% of herds, and 27.4%, 2.6%, 10.8% and 0.8% of faecal samples in 2015, while 100%, 0.0%, 86.7% and 60.0% of herds and 52.3%, 0.0%, 13.3% and 26.8% of faecal samples were positive in 2021/2022. Co-infections with two or more helminth taxa were detected in 74.4% of herds and 5.0% of faecal samples in 2015, and in 93.3% of herds and 21.7% of faecal samples in 2021/2022. The correlations between strongyle EPG, D. viviparus LPG and F. hepatica EPG were significantly positive in 2015. Significantly higher mean EPGs were identified in 2015 in faecal samples presenting co-infections with F. hepatica and one or two other helminth taxa than in faecal samples presenting F. hepatica mono-infections (P = 0.013). Although expected, the infection status (mono- or co-infected) had no significant impact on milk yield, milk protein and milk fat content in the linear mixed model analyses based on individual faecal examinations. Conclusions Patent helminth co-infections had no additive detrimental impact on milk production parameters in the present study. This might be a result of presumably low worm burdens, but should be confirmed in future studies. Graphical Abstract

Abstract Background Lymnaeid snails of the genus Austropeplea are an important vector of the liver fluke (Fasciola hepatica), contributing to livestock production losses in Australia and New Zealand. However, the species status within Austropeplea is ambiguous due to heavy reliance on morphological analysis and a relative lack of genetic data. This study aimed to characterise the mitochondrial genome of A. cf. brazieri, an intermediate host of liver fluke in eastern Victoria. Methods The mitochondrial genome was assembled and annotated from a combination of second- and third-generation sequencing data. For comparative purposes, we performed phylogenetic analyses of the concatenated nucleotide sequences of the mitochondrial protein-coding genes, cytochrome c oxidase subunit 1 and 16S genes. Results The assembled mt genome was 13,757 base pairs and comprised 37 genes, including 13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes. The mt genome length, gene order and nucleotide compositions were similar to related species of lymnaeids. Phylogenetic analyses of the mt nucleotide sequences placed A. cf. brazieri within the same clade as Orientogalba ollula with strong statistical supports. Phylogenies of the cox1 and 16S mt sequences were constructed due to the wide availability of these sequences representing the lymnaeid taxa. As expected in both these phylogenies, A. cf. brazieri clustered with other Austropeplea sequences, but the nodal supports were low. Conclusions The representative mt genome of A. cf. brazieri should provide a useful resource for future molecular, epidemiology and parasitological studies of this socio-economically important lymnaeid species. Graphical abstract

Background: The productivity of livestock production has been greatly impacted by parasitic diseases. The infection prevalence of parasites in native and imported slaughtered animals has not received much attention, despite the fact that Saudi Arabia has data on animal parasites. This study aimed to determine the infection prevalence of parasites in slaughtered animals in relation to host origin and host age. Methods: This investigation was conducted at Al Makhwah area in southwestern Saudi Arabia for six months from September 2021 to February 2022. The examined animals were 1208 in total, including goats (504), sheep (440) and cattle (264). All the animals examined were slaughtered at official abattoirs. The collected parasites were preserved, processed and identified. Result: Out of 1208 examined, 72 were infected (5.96%). The parasite community consisted of six species, Dicrocoelium dendriticum, Fasciola hepatica, F. giagantica, Monezia expansa, Echinococcus granulosus (Hydatiid cyst) and Haemonchus contortus. Hydatidosis was a more prevalent parasitic disease. The infection prevalence of hydatidosis in the examined animals was high in cattle (4.85%) while were 3.76 and 4.77% in goats and sheep respectively. Significant differences in prevalence per host age in goats, sheep and cattle were found. The prevalence of parasites in imported animals was higher (16.67, 17.5 and 11.36% in cattle, sheep and goats respectively) as compared to that in native ones (5.42, 6.5 and 3.69% in cattle, sheep and goats respectively). Based on our findings, Hydatidosis was identified as the most prevalent parasitic disease. Imported animals had a higher parasitic infection compared to local animals. It is essential to take necessary measures to prevent the transmission of Hydatidosis and improper organ condemnation from abattoirs. We recommend conducting further research to identify the reasons why imported animals are more likely to become infected than local animals. [ABSTRACT FROM AUTHOR]

We investigated herein the prevalence of Fasciola hepatica infection in sheep at Sejnane slaughterhouse, governorate of Bizerte, Northwest of Tunisia, using three different diagnostic techniques (liver dissection, bile examination, and coprology). Faeces, liver, gall bladder as well as blood samples were collected from 603 slaughtered sheep in two seasons: winter and summer. Faecal egg counts of F. hepatica were estimated using sedimentation technique. Livers were examined for the presence of flukes, and bile collected from gall bladder was examined by sedimentation technique for the presence of F. hepatica eggs. Faecal egg counts of gastrointestinal helminths were estimated using flotation followed by the McMaster technique. Blood samples were used to estimate blood cell count (RBC) (×106/mL), haemoglobin (Hb) (g/dL), and haematocrit (Ht) (%) levels. A total of 1714 F. hepatica flukes were collected from 68 infected livers, the number of flukes per sheep ranged between naught and 195. Bile examination (16.78% ± 1.83; 51/310) showed the higher infection prevalence, followed by liver dissection (11.28% ± 1.17; 68/603) and coprology (9.12% ± 1.08; 55/603) (p = 0.015). Infection prevalences were significantly higher in young sheep aged of less than 1 year (8.13% ± 1.22; 49/498), in cross‐bred sheep (10.61% ± 1.39%; 64/478), and in summer (7.13% ± 1.82; 43/293) (p < 0.05). There was no significant difference in infection prevalence by gastrointestinal helminths in F. hepatica‐infected and F. hepatica‐non‐infected animals (p > 0.05). The overall prevalence of F. hepatica‐infected anaemic sheep was higher (22.73% ± 4.47; 20/88) than F. hepatica‐non‐infected anaemic sheep (p < 0.05). Fasciola hepatica infection is frequent in sheep from Sejnane representing hence an important constraint for the development of the sheep industry in this region. Therefore, it is necessary to establish and implement a specific control programme to reduce fasciolosis infection risks including animal owners' education. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the prevalence and level of contamination of Naira notes in circulation in Ile Ife, Osun State, Nigeria. Nine hundred and sixty (960) Naira notes comprising all denominations were collected randomly from commercial bus drivers, point of sales (POS) operators, bread sellers and open market traders between October 2023 and March 2024. The parasite egg/cysts were recovered using a standard parasitological procedure. Two hundred and thirty-seven (237) Naira notes harboured one or more parasites giving an overall prevalence of 24.7%. Ten parasites were recovered which include Ascaris lumbricoides (14.1%) Taenia spp (7.8%), Enterobius vermicularis (5.9%), Entamoeba histolytica (3.4%), Trichuris trichiura (0.6%), Ancylostoma duodenale (5.0%), Giardia lamblia (1.3%), Fasciola hepatica (1.6%), Entamoeba coli (4.7%) and Balantidium coli (0.6%). The highest prevalence of contamination was recorded in N100 notes (60.0%) followed by N200 Naira notes (26.7%) while the joint lowest contamination rate of 10% was recorded in N10 Naira and N20 Naira notes respectively. Old notes were more contaminated (29.8%) than the newly redesigned notes (4.6%) and higher parasitic contamination was recorded among paper notes (27.7%) than the polymers (18%). Mutilated Naira notes were more contaminated (85.7%) than very dirty ones (50%). Based on the source of currency, Naira notes collected from commercial drivers had the highest contamination (38.8%). Statistical associations were recorded between parasitic contamination of currency and currency denomination, currency type, source of currency note, and physical condition of Naira note (P<0.05) except among currency materials where a nonsignificant association was recorded (P=0.05). This study has revealed that currency notes in circulation in Ile-Ife, are contaminated with a variety of parasites that pose a significant risk to public health and it is essential to promote good personal hygiene practices, such as washing hands regularly, especially after using the toilet and handling currency notes. [ABSTRACT FROM AUTHOR]

Fasciola hepatica, also known as the common liver fluke, is a globally distributed trematode parasite responsible for high economic losses in ruminants. Infection with F. hepatica occurs in Polish cattle and sheep; however, very little is known about its occurrence in goats. Therefore, a serological and coproscopic survey was carried out in Polish goats to determine the herd-level prevalence of F. hepatica infection in the goat population of Poland. Between 2014 and 2022, 33 randomly selected goat herds were serologically screened in the regions of Poland for which risk of F. hepatica infection was estimated as increased based on the spatial distribution model developed within the frame of the GLOWORM project. Virtually all adult goats (>1 year-old) were tested using a commercial MM3-SERO ELISA. Risk factors for seropositive herd status were analysed in contingency tables. Also, faecal samples from 214 goat herds monitored for gastrointestinal nematode infections and anthelmintic resistance were examined using a sedimentation method. At least one seropositive goat was detected in 11 of 33 herds, indicating herd-level seroprevalence of 33.3% (95% confidence interval (CI 95%): 19.7%–50.4%). At the animal level, only 17 of 1,464 tested goats were seropositive (1.2%, CI 95%: 0.7%–1.9%). The within-herd seroprevalence ranged from 0.8% to 11.1%. The serological status of the herd was not significantly associated with the characteristics of the herd or the extent of contact with sheep. In one herd, located in central Poland, a single positive faecal sample was found indicating a herd-level prevalence of F. hepatica infection of 0.5% (CI 95%: 0.1%–2.6%). The animal's post-mortem examination revealed liver lesions typical of chronic fasciolosis. F. hepatica infection occurs sporadically in Polish goat population and its prevalence is much lower than in cattle or sheep. Therefore, treatment or prevention of fasciolosis should only be considered if it has been reliably confirmed by an accurate diagnostic test. This applies also to goats inhabiting geographical areas where F. hepatica infection appears to be widespread in cattle and sheep, very likely due to the fact that goats avoid wet areas. [ABSTRACT FROM AUTHOR]