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Infection with Fasciola hepatica (liver fluke) causes fasciolosis (or fascioliasis) and poses a considerable economic as well as welfare burden to both the agricultural and animal health sectors. Here, we explore the ex vivo anthelmintic potential of synthetic derivatives of hederagenin, isolated in bulk from Hedera helix. Thirty-six compounds were initially screened against F. hepatica newly excysted juveniles (NEJs) of the Italian strain. Eleven of these compounds were active against NEJs and were selected for further study, using adult F. hepatica derived from a local abattoir (provenance unknown). From these eleven compounds, six demonstrated activity and were further assessed against immature liver flukes of the Italian strain. Subsequently, the most active compounds (n = 5) were further evaluated in ex vivo dose response experiments against adult Italian strain liver flukes. Overall, MC042 was identified as the most active molecule and the EC50 obtained from immature and adult liver fluke assays (at 24 h post co-culture) are estimated as 1.07 μM and 13.02 μM, respectively. When compared to the in vitro cytotoxicity of MDBK bovine cell line, MC042 demonstrated the highest anthelmintic selectivity (44.37 for immature and 3.64 for adult flukes). These data indicate that modified hederagenins display properties suitable for further investigations as candidate flukicides.

SARS-CoV-2, the causal agent of COVID-19, is a new coronavirus that has rapidly spread worldwide and significantly impacted human health by causing a severe acute respiratory syndrome boosted by a pulmonary hyperinflammatory response. Previous data from our lab showed that the newly excysted juveniles of the helminth parasite Fasciola hepatica (FhNEJ) modulate molecular routes within host cells related to vesicle-mediated transport and components of the innate immune response, which could potentially be relevant during viral infections. Therefore, the aim of the present study was to determine whether FhNEJ-derived molecules influence SARS-CoV-2 infection efficiency in Vero cells. Pre-treatment of Vero cells with a tegument-enriched antigenic extract of FhNEJ (FhNEJ-TEG) significantly reduced infection by both vesicular stomatitis virus particles pseudotyped with the SARS-CoV-2 Spike protein (VSV-S2) and live SARS-CoV-2. Pre-treatment of the virus itself with FhNEJ-TEG prior to infection also resulted in reduced infection efficiency similar to that obtained by remdesivir pre-treatment. Remarkably, treatment of Vero cells with FhNEJ-TEG after VSV-S2 entry also resulted in reduced infection efficiency, suggesting that FhNEJ-TEG may also affect post-entry steps of the VSV replication cycle. Altogether, our results could potentially encourage the production of FhNEJ-derived molecules in a safe, synthetic format for their application as therapeutic agents against SARS-CoV-2 and other related respiratory viruses.

Cysteine proteases are widespread in all life kingdoms, being central to diverse physiological processes based on a broad range of substrate specificity. Paralogous Fasciola hepatica cathepsin L proteases are essential to parasite invasion, tissue migration and reproduction. In spite of similarities in their overall sequence and structure, these enzymes often exhibit different substrate specificity. These preferences are principally determined by the amino acid composition of the active site's S2 subsite (pocket) of the enzyme that interacts with the substrate P2 residue (Schetcher and Berger nomenclature). Although secreted FhCL1 accommodates aliphatic residues in the S2 pocket, FhCL2 is also efficient in cleaving proline in that position. To understand these differences, we engineered the FhCL1 S2 subsite at three amino acid positions to render it identical to that present in FhCL2. The substitutions did not produce the expected increment in proline accommodation in P2. Rather, they decreased the enzyme's catalytic efficiency toward synthetic peptides. Nonetheless, a change in the P3 specificity was associated with the mutation of Leu67 to Tyr, a hinge residue between the S2 and S3 subsites that contributes to the accommodation of Gly in S3. Molecular dynamic simulations highlighted changes in the spatial distribution and secondary structure of the S2 and S3 pockets of the mutant FhCL1 enzymes. The reduced affinity and catalytic efficiency of the mutant enzymes may be due to a narrowing of the active site cleft that hinders the accommodation of substrates. Because the variations in the enzymatic activity measured could not be exclusively allocated to those residues lining the active site, other more external positions might modulate enzyme conformation, and, therefore, catalytic activity.

Liver fluke (Fasciola hepatica) is a parasite of herbivores including wildlife. Macropods, such as Eastern grey kangaroo (Macropus giganteus) and Common wallaroo (Osphranter robustus), are frequently observed sharing grazing sites with domestic livestock. The impact of Macropods, as reservoirs of infection, on livestock production and risks to cross-species transmission are largely unknown. In Phase 1 of this study, liver and faecal samples were collected from 245 Macropods (181 Eastern grey kangaroos, 64 Common wallaroos) cohabiting livestock farms (n = 7) in the Northern Tablelands regions of New South Wales. Total fluke (TFC) and fluke eggs (FEC) were counted in the liver and faeces, respectively, to assess prevalence. Faecal antigens were also measured using the commercial Bio-X Diagnostic Monoscreen AgELISA Fasciola hepatica kit (cELISA) to assess suitability as a diagnostic tool. In Phase 2, Macropod faecal samples were collected from 60 livestock farms to conduct FEC and assess prevalence by region. Liver fluke was prevalent in 22% of Eastern grey kangaroo and 20% of Common wallaroos with prevalence as high as 45% in the Eastern grey kangaroo. Fluke burdens ranged from 1 to 122 flukes (mean = 9 flukes) with a FEC range of 0–195 eggs per gram (epg) of faeces (mean = 18 epg). Evidence of dead and live flukes trapped within fibrotic capsules confirms the ability of Macropods to resolve infections. cELISA proved highly specific (100%) and sensitive (98%) in liver fluke detection however fibrotic capsules observed in the liver may reduce the correlation of coproantigens with fluke burden. Phase 2 revealed that 27% of livestock farms had Macropods infected with liver fluke. Overall, this study confirmed Eastern grey kangaroo and Common wallaroo are susceptible hosts and potential reservoirs for liver fluke and, monitoring infections in Macropods would assist in livestock disease management.

Fasciolosis caused by the liver flukes Fasciola hepatica and Fasciola gigantica is one of the most important neglected parasitic diseases of humans and animals. The ability of the parasites to infect and multiply in their intermediate snail hosts, and their adaptation to a wide variety of mammalian definitive hosts contribute to their high transmissibility and distribution. Within the mammalian host, the trauma caused by the immature flukes burrowing through the liver parenchyma is associated with most of the pathogenesis. Similarly, the feeding activity and the physical presence of large flukes in the bile ducts can lead to anemia, inflammation, obstruction and cholangitis. The high frequency of non-synonymous polymorphisms found in Fasciola spp. genes allows for adaptation and invasion of a broad range of hosts. This is also facilitated by parasite’s excretory-secretory (ES) molecules that mediate physiological changes that allows their establishment within the host. ES contains cathepsin peptidases that aid parasite invasion by degrading collagen and fibronectin. In the bile ducts, cathepsin-L is critical to hemoglobin digestion during feeding activities. Other molecules (peroxiredoxin, cathepsin-L and Kunitz-type inhibitor) stimulate a strong immune response polarized toward a Treg/Th2 phenotype that favors fluke’s survival. Helminth defense molecule, fatty acid binding proteins, Fasciola-specific glycans and miRNAs modulate host pro-inflammatory responses, while antioxidant scavenger enzymes work in an orchestrated way to deter host oxidant-mediated damage. Combining these strategies Fasciola spp. survive for decades within their mammalian host, where they reproduce and spread to become one of the most widespread zoonotic worm parasites in the world.

Fasciola hepatica is a zoonotic liver trematode that usually causes infection in cattle and sheep, and is transmitted to humans by consuming water and aquatic plants contaminated with metacercaria. The detection of Fasciola eggs in stools, serological evaluation and radiological evaluation are essential for diagnosis. Triclabendazole is the first-line therapy for fascioliasis. However, as triclabendazole is not an easily accessible drug in countries such as Turkey, it reveals a quest for alternative therapies. In this report, we present a 10-year-old boy with fascioliasis successfully treated with a course of metronidazole 1.5 g/ day for 3 weeks in 2020. During the follow-up, eosinophilia and radiological findings completely recovered. Here we report a case of pediatric fascioliasis that was cured with metronidazole successfully.

IntroductionFascioliasis is a parasitic foodborne disease caused by the liver flukes, Fasciola hepatica and F. gigantica. Such parasites cause serious illness in numerous domestic animals and also in humans. Following infection, the parasite secretes a variety of molecules that immediately interact with the host immunity to establish successful infection. These molecules include cathepsin L peptidase 1 (CatL1); the highly investigated diagnostic and vaccine antigens using various animal models. However, a few studies have analyzed the potentials of FhCatL1 as a diagnostic or vaccine antigen using bioinformatic tools and much less for FgCatL1. The present study provides inclusive and exclusive information on the physico-chemical, antigenic and immunogenic properties of F. hepatica cathepsin L1 (FhCatL1) protein using multiple bioinformatic analysis tools and several online web servers. Also, the validation of our employed available online servers was conducted against a huge collection of previously published studies focusing on the properties of FhCatL1as a diagnostic and vaccine antigen.MethodsFor this purpose, the secondary, tertiary, and quaternary structure of FhCatL1 protein were also predicted and analyzed using the SWISS-MODEL server. Validation of the modeled structures was performed by Ramachandran plots. The antigenic epitopes of the protein were predicted by IEDB server.Results and discussionOur findings revealed the low similarity of FhCatL1 with mammalian CatL1, lacking signal peptides or transmembrane domain, and the presence of 33 phosphorylation sites. Also, the containment of FhCatL1 for many topological, physico-chemical, immunological properties that favored its function of solubility and interaction with the immune components were reported. In addition, the earlier worldwide reports documented the high efficacy of FhCatL1 as a diagnostic and vaccine antigen in different animals. Altogether, FhCatL1 is considered an excellent candidate for using in commercialized diagnostic assays or vaccine products against fascioliasis in different animal species. Our assessment also included FgCatL1 and reported very similar findings and outputs to those of FhCatL1.

Among the gastrointestinal parasitoses, fascioliasis is one of the most common diseases in ruminants. Fasciola spp. is recorded on five continents of the globe, in more than 50 countries. The parasitizing trematode causes economic losses associated with a decrease in milk yield, body weight, and culling of affected carcasses and organs. In this study, we aimed to quantify the prevalence of fascioliasis among ruminants (cattle, sheep and goats) of the world in terms of the odds ratio according to the Mantel–Haenszel test (M–H). Online databases in English, Russian and Ukrainian languages were searched for publications from January 2002 to September 2020. This meta-analysis included 42 studies with ruminant hosts. Data on the spread of fascioliasis were collected from different continents of the globe: North and South America, Europe, Asia and Africa. The results of the study found that the overall prevalence of fascioliasis in cattle was 6.41 %, while in small ruminants it was only 2.03 %. The disease in cattle was recorded 1.48 times more often than in sheep and goats. Egger’s regression test revealed no significant publication bias (P = 0.265). The results of the meta-analysis confirm that the causative agent of fascioliasis circulates mainly in the emerging countries. The updated data on fascioliasis will expand the screening strategy to maintain the health of farm ruminants and reduce economic losses.

For the first time, historical and recent records on fascioliasis in Egypt reveal the results of sympatric and allopatric fluke/snail interactions in the Manzala Lagoon area by exploring the susceptibility of the species Pseudosuccinea columella after exposure to the Fasciola hepatica isolates from Egypt. Many factors influence trace metal accumulation in Pseudosuccinea columella, including temperature and salinity. Meanwhile, other factors are important in decreasing the bioaccumulation of heavy metals as low pH, low temperature, and high organic content of the substrate. It is indicated, from this study, that the Fe and Cu contamination can be detected in Pseudosuccinea columella species. The abundance of these snails varied all over the seasons where the minimum value was recorded in summer while reaching the maximum in autumn. Moreover, the abundance increased with increasing rainfall and humidity during the winter season.

Fasciola hepatica is a trematode parasite that infects animals and humans causing fasciolosis, a worldwide-distributed disease responsible for important economic losses and health problems. This disease is of growing public health concern since parasite isolates resistant to the current treatment (triclabendazole) have increasingly been described. F. hepatica infects its vertebrate host after ingestion of the encysted parasite (metacercariae), which are found in the water or attached to plants. Upon ingestion, newly excysted juveniles of F. hepatica (FhNEJ) emerge in the intestinal lumen and cross the intestinal barrier, reach the peritoneum and migrate to the biliary ducts, where adult worms fully develop. Despite the efforts made to develop new therapeutic and preventive tools, to date, protection against F. hepatica obtained in different animal models is far from optimal. Early events of host-FhNEJ interactions are of paramount importance for the infection progress in fasciolosis, especially those occurring at the host-parasite interface. Nevertheless, studies of FhNEJ responses to the changing host environment encountered during migration across host tissues are still scarce. Here, we set-up an ex vivo model coupled with quantitative SWATH-MS proteomics to study early host-parasite interaction events in fasciolosis. After comparing tegument and somatic fractions from control parasites and FhNEJ that managed to cross a mouse intestinal section ex vivo, a set of parasite proteins whose expression was statistically different were found. These included upregulation of cathepsins L3 and L4, proteolytic inhibitor Fh serpin 2, and a number of molecules linked with nutrient uptake and metabolism, including histone H4, H2A and H2B, low density lipoprotein receptor, tetraspanin, fatty acid binding protein a and glutathione-S-transferase. Downregulated proteins in FhNEJ after gut passage were more numerous than the upregulated ones, and included the heath shock proteins HSP90 and alpha crystallin, amongst others. This study brings new insights into early host-parasite interactions in fasciolosis and sheds light on the proteomic changes in FhNEJ triggered upon excystment and intestinal wall crossing, which could serve to define new targets for the prevention and treatment of this widespread parasitic disease. Author summary: Fasciolosis caused by the helminth parasite Fasciola hepatica is a serious health and economic problem worldwide. Treatment and prevention of this disease pose several drawbacks that have so far not been solved. The definition of suitable parasite molecular targets to overcome such drawbacks should be based on thoroughly deciphering host-parasite interactions, and in this regard most studies have focused on the adult stages of F. hepatica. Nevertheless, in this context, the study of the transient juvenile stages of this parasite could be of higher utility due to the importance of early interactions with the host for parasite migration and the successful establishment of infection. In this work, we set-up an ex vivo model and performed a quantitative proteomics approach to study the changes in F. hepatica juveniles upon gut passage. We found that the parasite tegument and somatic compartments experienced deep changes in their composition and showed that the host triggers the expression of specific molecules that are important for parasite migration and survival at this stage. The molecules described here could serve to better understand host-parasite interactions and to define new targets against fasciolosis. [ABSTRACT FROM AUTHOR]

Hepatic trematodes, such as Fasciola hepatica and Fascioloides magna, have variable distribution throughout the United States. F. magna is endemic in the upper midwestern United States, and F. magna infections are diagnosed frequently in weaned calves and adult beef cattle at the North Dakota State University Veterinary Diagnostic Laboratory (NDSU-VDL). Rarely, liver fluke infestation has also been observed in much younger calves, including aborted fetuses. We describe here, in 2 fetal and 7 neonatal beef calves submitted to the NDSU-VDL between 2011 and 2020, parasitic migration tracts in livers, consisting of regionally extensive, random, linear tracts of fibrosis admixed with black porphyrin pigment, along with foci of necrosis and hemorrhage, and mixed inflammatory cells, which were caused presumptively by F. magna infection. Samples were not available from our 9 cases for PCR assay and sequencing, but we did confirm F. magna within liver samples collected from regional cattle in 2020 and 2021. Fetal and neonatal trematodosis was often concurrent with other common causes of fetal abortion and neonatal calf loss in our cases; however, based on the prepatent period of F. magna, fetal and neonatal beef calf trematode infestations occurred in utero. [ABSTRACT FROM AUTHOR]

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Among the gastrointestinal parasitoses, fascioliasis is one of the most common diseases in ruminants. Fasciola spp. is recorded on five continents of the globe, in more than 50 countries. The parasitizing trematode causes economic losses associated with a decrease in milk yield, body weight, and culling of affected carcasses and organs. In this study, we aimed to quantify the prevalence of fascioliasis among ruminants (cattle, sheep and goats) of the world in terms of the odds ratio according to the Mantel-Haenszel test (M-H). Online databases in English, Russian and Ukrainian languages were searched for publications from January 2002 to September 2020. This meta-analysis included 42 studies with ruminant hosts. Data on the spread of fascioliasis were collected from different continents of the globe: North and South America, Europe, Asia and Africa. The results of the study found that the overall prevalence of fascioliasis in cattle was 6.41 %, while in small ruminants it was only 2.03 %. The disease in cattle was recorded 1.48 times more often than in sheep and goats. Egger's regression test revealed no significant publication bias (P = 0.265). The results of the meta-analysis confirm that the causative agent of fascioliasis circulates mainly in the emerging countries. The updated data on fascioliasis will expand the screening strategy to maintain the health of farm ruminants and reduce economic losses. [ABSTRACT FROM AUTHOR]

Fasciola hepatica, commonly referred to as liver flukes, is a substantial zoonotic parasitic disease of humans and livestock globally. While infection is readily controlled by anthelmintics, namely triclabendazole, the heavy reliance on triclabendazole has resulted in drug resistance appearing worldwide. Due to drug resistance, it is imperative to adopt an integrated parasite management program to preserve the efficacy of currently available anthelmintics. A integrated liver fluke management plan would benefit from a simple rapid, field-deployable diagnostic for detection of F. hepatica in environment and the host. Therefore, a rapid DNA test using loop-mediated isothermal amplification was developed and optimised for the detection of F. hepatica from faecal and water samples to enable the detection of parasites both within the host and from the environment. The assay presented here is fast, with amplification in ≤20 min, and highly sensitive, with a detection limit of 5 × 10-4 ng/µL. The workflow presented here provides a time to result of ≤60 min without requiring a commercial kit for the extraction of DNA from faecal and water samples, and pending further validation from field-samples, could potentially be used to enable real-time decision making to mitigate parasite prevalence on a farming property and with no requirement for sample transportation. [ABSTRACT FROM AUTHOR]

Background: In Saudi Arabia, more than US$ 0.2 million annual losses are caused by liver condemnations due to fascioliasis. Data obtained from the genetic characterization of Fasciola population sheds light on parasite transmission which could eventually help in development of effective parasite control measures. So, the aim of this study was to investigate the genetic diversity of Fasciola spp. isolated from cattle in Saudi Arabia by sequence analyses of COI gene. Materials and Methods: A total of 325 cows slaughtered at the central municipal abattoir in Jeddah city, Jeddah Province, Saudi Arabia were examined for fascioliasis in the period from 1st of June to 1st of July 2020. DNA was extracted from adult Fasciola worms and used for PCR and DNA sequence using a primer pair targeting COI gene. Analysis of the obtained sequences was done using BLAST search and phylogenetic analysis. Results: Bovine fascioliasis was diagnosed in 18 out of 325 cattle (5.5%). Forty-eight flukes were extracted from infected animals and DNA was successfully amplified from all flukes. Overall 12 different DNA sequences were obtained. BLAST search showed that all obtained sequences were F. hepatica and had > 97% similarity with F. hepatica isolates from Tanzania, Europe and Iran. Phylogenetic analysis of the obtained sequences showed that Fasciola isolates from the current study were clustered in one subclade closely related to isolates from North and South Africa and Italy. Conclusion: Reports on the molecular characterization of Fasciola spp. in Saudi Arabia are limited. In the current study, our findings showed that F. hepatica was the only Fasciola species parasitizing cattle in Jeddah city, Saudi Arabia. Further studies using a large number of samples from different localities in Saudi Arabia are needed to provide data that will help the development of control measures against fascioliasis. [ABSTRACT FROM AUTHOR]

In the study, the level of antibodies to fasciolas in the milk of cows in the Moscow region of Russia was assessed and the relationship between their number and the productive characteristics of these animals was traced. In October 2021, a milk sample was taken from a dairy farm in the Moscow region of Russia, and the number of antibodies to Fasciola hepatica (ODRf) and Ostertagia ostertagi (ODRo) was assessed. Their level was determined using multivariate linear regression models and their relationship with economically significant indicators was traced: the amount of protein in milk, the percentage of fat in milk, and the interval between past calving. An increase in the ODRf quartile from 25% (0.412) to 75% (0.976) was associated with a decrease in the volume of milk produced per year by 0.8 kg (day of the lactation process) (p=0.002), with a decrease in fat content in milk by 0.07 % (p<0.001), with an increase in the interval between sections by 4.6 days (p=0.04) in the absence of a significant relationship with the level of protein in milk. In the case of simultaneous infection with both parasites, the effect of ODRf and ODRo on the volume of milk produced was additional, rather than synergistic. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is an important pathogen that causes liver fluke disease in definitive hosts such as livestock animals and humans. Various excretory/secretory products have been used in serological diagnosis and vaccination studies targeting fasciolosis. There are no commercial vaccines against fasciolosis yet. Bioinformatic analysis based on computational methods have lower cost and provide faster output compared to conventional vaccine antigen discovery techniques. The aim of this study was to predict B- and T-cell specific epitopes of four excretory/secretory antigens (Kunitz-type serine protease inhibitor, cathepsin L1, helminth defense molecule, and glutathione S-transferase) of Fasciola hepatica and to construct a multiepitope vaccine candidate against fasciolosis. Initially, nonallergic and the highest antigenic B- and T- cell epitopes were selected and then, physico-chemical parameters, secondary and tertiary structures of designed multiepitope vaccine candidate were predicted. Tertiary structure was refined and validated using online bioinformatic tools. Linear and discontinuous B-cell epitopes and disulfide bonds were determined. Finally, molecular docking analysis for MHC-I and MHC-II receptors was performed. This multi-epitope vaccine candidate antigen, with high immunological properties, can be considered as a promising vaccine candidate for animal experiments and wet lab studies. [ABSTRACT FROM AUTHOR]

Two controlled efficacy studies were conducted to determine the effective dose of fosfatriclaben (FTCB) and compare its fasciolicidal efficacy with that of three commercial products against eggs and adult stages of Fasciola hepatica in artificially infected cattle. In study 1, 20 trematode-free Holstein Friesian steers were infected on day 0 with 500 F. hepatica metacercariae. Ten weeks after infection and the steers were confirmed to be positive for trematode eggs through a modified sedimentation method. On day 75, they were divided into five groups of four animals each for treatment. Group 1 (G1) served as the untreated control; G2, G3, and G4 received FTCB at 4, 6, and 8 mg/kg/ intramuscularly (IM), respectively. G5 received a combined treatment of triclabendazole (TCBZ) (12 mg/kg IM + ivermectin (0.2 mg/kg IM). Individual faecal analyses were performed on days -8, 0, 70, 75, and 105 to evaluate the reduction in trematode eggs. Four weeks after treatment, the steers were humanely slaughtered to harvest the livers and remove the parasites present in the bile ducts. Efficacy was evaluated by the reduction in fecal egg counts or in number of adult parasites, compared to the untreated control. The effective FTCB dose was 6 mg/kg. Once the effective dose was determined, study 2 was conducted on another 20 steers infected with 500 F. hepatica metacercariae, to compare the effectiveness of FTCB with three commercials fasciolicides. All procedures were performed as described in study 1, and treatments were as follows: Group 1 (G1), closantel (5 mg/kg subcutaneously (SC)); G2, TCBZ (12 mg/kg IM) + ivermectin (0.2 mg/kg IM); G3, FTCB (6 mg/kg IM); G4, triclabendazole (12 mg/kg) + albendazole (5 mg/kg/PO (orally); and G5 served as an untreated control. The results indicated that all tested compounds were highly effective in the reduction of faecal egg excretion (99.7-100%) and adult parasites (98.9-100%), except closantel, which exhibited low efficacy (74.4%) when tested against adult trematodes. We concluded that the effective dose of FTCB for cattle was 6 mg/kg IM, which is half the recommended clinical dose of the commercial combination of TCBZ and ivermectin. The fasciolicidal efficacy of FTCB was like the other three flukicides in reducing adult F. hepatica and Fasciola eggs; however, closantel was not sufficiently efficient against adult flukes., Competing Interests: Declaration of Competing Interest The authors declare that no conflict of interest exists, therefore we have nothing to declare., (Copyright © 2024. Published by Elsevier Inc.)

In many regions of New Zealand liver fluke is endemic, infecting most grazing ruminants, including cattle, sheep, and deer. Restricting the economic losses and welfare costs associated with liver fluke relies on accurately identifying those animals with a production limiting infection. This has proven a difficult goal and although several antemortem quantitative tests are available, including faecal egg counts (FEC), serum ELISA and copro-antigen ELISA, none can be considered a gold standard test of liver fluke infection. The accepted gold standard test for fascioliasis is the total fluke count, which is both laborious and can only be completed at post-mortem. This study aimed to compare the performance of four liver fluke diagnostic tests, against the results of a gold standard total fluke count test. Two groups of cattle were selected, 29 culled mixed age beef cows (MAC) and ten 30-month-old steers. The cattle were blood sampled and faecal sampled prior to slaughter and their whole livers recovered post slaughter at the abattoir. Liveweight was also recorded at slaughter. After collection, each liver was weighed, scored for gross pathology, then serum, faeces and livers were frozen at -20 °C for later analysis. Faecal egg counts and F. hepatica copro-antigen ELISA tests were completed on the faecal samples and total fluke counts were completed on the livers. Fasciola hepatica antibody concentration in serum samples were quantified using a commercial ELISA test. Poisson regression models were built to model the association between each diagnostic test and the total fluke count, and a linear regression model was built to examine the relationship between each diagnostic test and live weight at slaughter. The median fluke count was significantly higher in MAC than steers (p = 0.01), and F. hepatica eggs were present in 100% steers and 66% MAC. There was a significant effect of copro-antigen ELISA value on total fluke count (p < 0.0001), with a coproantigen ELISA value = 20.1 predicting 10 flukes and a value = 44.8 predicting 30 flukes. There was also a significant effect of FEC on total fluke count (p = 0.002) but the R-squared value for this model was lower. There was no association between liver fibrosis score or antibody ELISA test and total fluke count (p = 0.95, p = 0.73, respectively). There was a significant effect of total fluke count (p = 0.03) on liveweight at slaughter, with liveweight falling 20.4 kg for each unit increase in log e (total fluke count). There was no effect of FEC (p = 0.11), antibody ELISA (p = 0.55) or copro-antigen ELISA value (p = 0.16) on liveweight at slaughter. Taken together, these results show that the coproantigen ELISA test is the better test for estimating the true liver fluke burden and that the number of flukes in the liver has a negative effect on cattle live weights at slaughter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

During a survey for helminths in reindeer ( Rangifer tarandus ) across the Palearctic region, eggs were found in zoo reindeer feces. These were identified as eggs of Fasciola hepatica based on their morphology, morphometrics, location, and analysis of their partial sequence of ITS rDNA region. Some of the eggs had an appendage, previously unreported. Additionally, adults of F. hepatica were studied. Eggs derived from their uteri were also appendaged. Diagnostic morphological traits of F. hepatica eggs (abopercular appendage, knob, egg shell thickening, and egg shape) are discussed in this article. Three dimensional models of F. hepatica eggs were created to demonstrate the eggs features as best as possible. Since fecal examination remains gold standard in diagnosing fasciolosis in humans and animals worldwide, our findings may contribute to improved diagnostics. This research has also shown that reindeer can be a final host for F. hepatica . We also discuss whether the Novaya Zemlya archipelago might be the northernmost site of fasciolosis., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Background & Objective: Infestation of humans with helminth parasites can reduce the severity of some auto-inflammatory symptoms in humans. This study was done to evaluate the impact of the somatic antigens extracted from Fasciola hepatica (as an indicator Trematode) ,and Teladorgasia circumcincta (as an indicator nematode) on the immune responses of NMRI mice challenged with sheep red blood cells (SRBCs). Materials & Methods: The mice in treatment groups were intraperitoneally immunized with 1×109 SRBCs twice with 14 days intervals. Concurrent with the immunization, the mice received the extract of each of the parasites (50,100. And 150 µg of protein) or placebo, throughout the study on a daily basis. The specific cellular immune responses and the anti-SRBC antibody titers were detected by footpad thickness and, microhemagglutination test, respectively. Splenocytes were also monitored for cytokine production, proliferation rate, and respiratory burst. Results: The extracts of F. hepatica and T. circumcincta had an opposite effect on the change of the Anti-sRBC antibody level. The extract of F. hepatica caused a significant decrease in the antibody level whereas, extract T. circumcincta did not show any significant changes in the anti-SRBC antibody. Both extracts caused a significant decrease in the level of delayed-type hypersensitivity. However, the extract of F. hepatica caused a more profound reduction in the severity of delayed-type hypersensitivity. The level of IFN-γ in the splenocytes of immunized mice receiving the F. hepatica showed a more pronounced decrease than the immunized mice receiving the extract of T. circumcincta extract. IL-10 levels were only increased in the immunized mice that received the extract of F. hepatica. Conclusion: The extract of F. hepatica may have an immunosuppressive property, while the extract of T. circumcincta may have immunomodulatory properties. Keywords:

The purpose of the research is to study triclabendazole effects on the Fasciola’s life-supporting organs, spines and suckers which are responsible for stable position of the parasite in the host.Materials and methods. The study material was trematodes Fasciola hepatica (Linneus 1758, family Fasciolidae Railliet 1895), which were collected after the action of triclabendazole (fasinex) (chemically 5-chloro-6-(2,3-dichlorophenoxy)-2-methylthiobenzimidazole)on the 7th day after the drug administered at a single dose of 10 mg/kg for the Active Substance in the treatment of ovine fasciolosis. F. hepatica from untreated animals served as control. Mature F. hepatica collected after treatment with triclabendazole, and marita from the control groups were dehydrated in ascending alcohol series for 1–2 days after fixation; then passed through a mixture of chloroform and absolute alcohol (in a ratio of 1:1), and through pure chloroform in two portions for 10–15 minutes. The material was then soaked in a mushy mixture of chloroform and paraffin in a thermostat at 37 °C for 12–18 hours, and in paraffin in a thermostat at 56 °C for 30–45 minutes; and then embedded in paraffin with added wax. The resulting paraffin blocks were broken down into serial sections of 5–7 μm thick, then stained and examined under a light microscope.Results and discussion. Pathomicromorphological analysis of F. hepatica’s spines and suckers, organs that come into adhesive contact with the host organism revealed destructive changes in them after the action of triclabendazole. After the action of triclabendazole on fascioles, the spines look enlarged and swollen, and have a more rounded shape and some changes in color, absorbing eosin in greater concentration. The muscle fibers of the fascioles’ oral and abdominal suckers also look swollen after the action of triclabendazole. Although the musculature of the F. hepatica’s pharynx retained its structure, it has changes. It thickened sharply, which is clearly visible on the transverse and longitudinal sections of the helminths; neurosecretory cells are destroyed, and voids are observed in their place.

Liver trematodes, including Fasciola and Dicrocoelium species, are zoonotic parasites that cause high mortality and economic losses in ruminants. This study aimed to investigate the effect of age, sex, and season on the prevalence of fasciolosis and dicrosliosis in slaughtered animals in the Tabriz slaughterhouse. During two years, livers of 4150 cows, 500 buffaloes, 2000 goats, and 5000 sheep were examined for the presence of Fasciola hepatica, Fasciola gigantica, and Dicrocoelium dendriticum by incisions on the liver and ocular observation. The results showed that 5.4%, 16%, and 6% of slaughtered cows, sheep, and goats were infected with liver trematodes (P

We conducted a retrospective cohort study of children who had chronic fascioliasis in the highlands of Peru to determine triclabendazole treatment efficacy. Children passing Fasciola eggs in stool were offered directly observed triclabendazole treatment (>1 doses of 10 mg/kg). Parasitologic cure was evaluated by using microscopy of stool 1–4 months after each treatment. A total of 146 children who had chronic fascioliasis participated in the study; 53% were female, and the mean ± SD age was 10.4 ± 3.1 years. After the first treatment, 55% of the children achieved parasitologic cure. Cure rates decreased after the second (38%), third (30%), and fourth (23%) treatments; 17 children (11.6%) did not achieve cure after 4 treatments. Higher baseline egg counts and lower socioeconomic status were associated with triclabendazole treatment failure. Decreased triclabendazole efficacy in disease-endemic communities threatens control efforts. Further research on triclabendazole resistance and new drugs to overcome it are urgently needed.

Abstract Fascioliasis is a parasitic infection caused by Fasciola spp. in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validate methods for the diagnosis of fascioliasis in animals. This study aimed to compare the loop‐mediated isothermal amplification (LAMP) technique with PCR assay for the diagnosis of F. hepatica in sheep. In this cross‐sectional study, 195 stool samples were collected from sheep for 3 months in Lorestan province, West of Iran. Specimens’ parasitological examination was performed by using the direct wet mount and formalin‐ether concentration method. After DNA extraction from the samples, molecular analysis was done using PCR and LAMP techniques based on the Fasciola ribosomal intergenic spacer (IGS) sequence. Of 195 specimens of sheep, 11 specimens were identified as F. hepatica‐positive infection by using microscopic, PCR and LAMP assays. Kappa agreement test results showed that there was a significant agreement between the results of microscopic examination diagnostic tests, PCR and LAMP (Kappa = 0.51–0.72 and p

Background and Aim: Human fascioliasis is a water and food-borne parasitic disease and has become endemic in the coastal provinces of the Caspian Sea. Limited reports of the disease and its diagnosis have resulted in ignoring the disease and a significant rise in the number of neglected patients in other parts of the country. Case presentation: In this study, a 50-year-old man living in a village of Kamyaran, in Kurdistan Province in Iran was referred to a hospital with complaints of weight loss, epigastric pain, and anorexia from two months ago. The patient-reported history of consuming raw vegetables and plant foods. Laboratory studies revealed anemia, leukocytosis, eosinophilia, high ESR, low serum iron, and elevated liver enzymes. A Spiral CT scan of the liver showed hypodense areas without enhancement in the arterial phase and with relatively homogeneous enhancement in the delayed phase. Ultrasound studies showed multiple hetero and hyper-echoic areas without walls in different parts of the liver. Finally, Fasciola hepatica was diagnosed after stool examination by formalin ether concentration method and also positive ELISA test. The patient was treated with triclabendazole 500 mg/day for two days. In the follow-up, the clinical status of the patient was better and we found improved laboratory test results. Conclusion: The present report highlights the need for paying more attention to the occurrence of this disease in Kurdistan Province by health care systems. Human fascioliasis, Fasciola hepatica, Kurdistan, Iran

Fasciola hepatica, commonly referred to as liver flukes, is a substantial zoonotic parasitic disease of humans and livestock globally. While infection is readily controlled by anthelmintics, namely triclabendazole, the heavy reliance on triclabendazole has resulted in drug resistance appearing worldwide. Due to drug resistance, it is imperative to adopt an integrated parasite management program to preserve the efficacy of currently available anthelmintics. A integrated liver fluke management plan would benefit from a simple rapid, field-deployable diagnostic for detection of F. hepatica in environment and the host. Therefore, a rapid DNA test using loop-mediated isothermal amplification was developed and optimised for the detection of F. hepatica from faecal and water samples to enable the detection of parasites both within the host and from the environment. The assay presented here is fast, with amplification in ≤20 min, and highly sensitive, with a detection limit of 5 × 10−4 ng/µL. The workflow presented here provides a time to result of ≤60 min without requiring a commercial kit for the extraction of DNA from faecal and water samples, and pending further validation from field-samples, could potentially be used to enable real-time decision making to mitigate parasite prevalence on a farming property and with no requirement for sample transportation.

Fasciola hepatica is reported as a trematode of great importance, causing fasciolosis, one of the most neglected zoonotic parasitic diseases in the world. In Brazil, potential losses of around US$ 210 million per year were calculated, with bovine fasciolosis alone. The State of Santa Catarina is located in the South Region of Brazil, with a herd of more than 4 million ruminants. The Serrana Mesoregion, historically free of fasciolosis, is composed of 30 municipalities, which have a consolidated agricultural tradition, mainly in extensive ruminant livestock. The objective of the present work was to investigate the occurrence and distribution of autochthonous cases of bovine fasciolosis in the region. For this, different analyzes were carried out in rural properties from eight municipalities, and in three slaughterhouses under the State Inspection Service (SIE), which received animals from all the Serrana Mesoregion. The rural properties were randomly selected, and up to 20 fecal samples were collected from random cattle older than 1 year, for analysis by coproparasitological sedimentation test. The traceability records of the positives were checked, for identifying autochthonous cases. Additionally, the tracking data of cattle slaughtered between the years of 2018 and 2021, which presented liver condemnations, were analyzed for defining autochthonous cases of bovine fasciolosis. In total, there were fecal samplings in 106 farms, with 1,927 tested cattle. From these, 178 (9.2%) were positive and autochthonous, but the overall result did not reflect the heterogeneity found between the evaluated municipalities. During the analyzed period, 11,556 cattle were slaughtered, with 1,744 (15.1%) liver condemnations due to fasciolosis in animals that never left the Serrana Mesoregion. The present study confirmed the presence of autochthonous cases of bovine fasciolosis in municipalities of the Serrana Mesoregion, and verified a high rate of positivity in animals slaughtered in the regional slaughterhouses. When the prevalence of bovine fasciolosis in vivo was compared with the postmortem positivity index in five municipalities, the results disagreed in four municipalities (p-value < 0.05), emphasizing the need for field investigation for the better understanding of the distribution and frequency of the parasitosis.

Human hepatic fasciolosis has been reported in 81 countries, some of which are endemic areas. In Europe, case reports from humans were published in Portugal, Spain, France, and Italy. Regarding Romania, we do not have any data on the prevalence of this parasitosis, with the exception of two cases of twins that were born in Romania and diagnosed in the last 37 years in Italy after joining their mother that lived there. We report the case of a patient diagnosed in Romania with chronic fasciolosis, presented as a hepatic pseudotumor that was diagnosed during the histopathological examination of the hepatic lesion. The patient received oral treatment with triclabendazole, two doses of 10 milligrams (mg) per kilogram (kg) of body weight, given 12 h apart, with no side effects during or after the treatment. The evolution of the patient was favorable. In conclusion, even in areas free of human fasciolosis, the presence of an anemic syndrome especially in children, abdominal pain in the upper quadrants, associated or not with other digestive manifestations, even more so associated with eosinophilia in the acute phase, should be carefully evaluated for ruling out a parasitosis such as fasciolosis even in countries where this diagnosis seems unlikely.

One of the rare manifestations of fascioliasis is liver abscess. In this paper, we report the case of a 38-year-old woman with a liver abscess caused by Fasciola hepatica (F. hepatica). The patient was referred to the clinic with recurrent fever, right upper quadrant (RUQ) pain, and a large abscess in her liver. Despite the consumption of an antibiotic drug, she still had symptoms. The symptoms began to disappear upon starting the consumption of triclabendazole (TCBZ). Fascioliasis can manifest itself with unusual symptoms that provide no specific clue for its diagnosis. Therefore, it is important to consider F. hepatica in the differential diagnosis of liver abscess, especially in endemic regions. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is a neglected parasitic infection with significant human health and livestock industry impact. The Andean Altiplano harbors an estimated 50% of the Fasciola's world infection burden. There is scarce data regarding the spatial associations between different Fasciola hosts. In this project, we aimed to determine the geospatial relationships between Fasciola eggs passed in feces of different livestock species and the risk of infection among each household as a unit. We used data from a cross-sectional study evaluating children and livestock feces for Fasciola infection around households in three districts of Anta province, in the Cusco region of Peru. Each sample was geographically tagged and evaluated for fascioliasis using microscopy methods. A total of 2070 households were included, the median age was 9.1 years (6.7–11.8), 49.5% were female, and 7.2% of the households had at least one infected child. A total of 2420 livestock feces samples were evaluated. The infection rate in livestock samples was 30.9%. The highest infection rate was found in sheep with 40.8%, followed by cattle (33.8%), and swine (26.4%). The median distance between a household with an infected child to a positive animal sample was 44.6 meters (IQR 14.7–112.8) and the distance between a household with no infected children to a positive animal sample was 62.2 meters (IQR 18.3–158.6) (p = 0.025). The multivariable logistic regression adjusted by presence of poor sanitation, unsafe water consumption, altitude, and presence of multiple infected children per household demonstrated an association between household infection and any cattle feces at a 50 meters radius (Uninfected: OR 1.42 (95%CI 1.07–1.89), p = 0.017. Infected: OR 1.89 (95%CI 1.31–2.73), p = 0.001), positive cattle feces at a 100 meters radius (OR 1.35 (95% CI 1.08–1.69), p = 0.008), and negative cattle feces at a 200 meters radius (OR 1.08 (95% CI 1.01–1.15), p = 0.022). We identified potential hot and cold spots for fascioliasis in the Anta province. An association between environmental contamination with feces from different livestock species and infected children in rural households was found in our study. Local health authorities may apply this strategy to estimate the risk of infection in human populations and apply targeted interventions. Author summary: Fasciola hepatica is the parasite that causes fascioliasis in the Andes region. The factors associated with transmission of the disease among humans have not been well described in the area studied. In this study, we evaluate the spatial relationships between infected households and the environmental burden of Fasciola eggs passed by different livestock in the Cusco region of Peru. We geographically marked and tested for fascioliasis feces provided by children and livestock. Infected and uninfected cattle as well as sheep feces were more likely to be in proximity to human households with infected children than to households without infected children. We were able to identify and map the areas of higher and lower infections rates. This information and strategy may be used in the control of the disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Fasciolosis caused by Fasciola hepatica is a serious disease and a huge challenge in the sheep industry. The disease has several clinical manifestations including acute death, anemia, ill-thrift and loss of body condition. Climate change with milder temperatures and heavier rainfall will increase the risk of fasciolosis. Grazing management and treatment with flukicide are at present the only options to restrain F. hepatica infection. However, control possibilities are challenging, and resistance to flukicide drugs is increasing. Diagnostic improvements, targeted treatment and vaccines will hopefully increase animal health and welfare on fluke infested pastures in the future. The liver fluke Fasciola hepatica may cause severe infection in several mammalian species, including sheep and humans. Fasciolosis is a parasitic disease occurring worldwide in temperate climates and involves intermediate lymnaeid snails as vectors, in Europe the pond snail Galba truncatula in particular. In the sheep industry, the disease is a serious welfare and health problem. Fasciolosis is usually classified as acute, subacute or chronic according to the number and stage of flukes present in the liver, but with a considerable overlap. Acute disease, associated with a large number of migrating larvae, often results in sudden death due to acute and massive hemorrhage, while chronic fasciolosis is characterized by anemia, hypoalbuminaemia and weight loss. The management of fasciolosis is an increasing challenge in the sheep industry. Early diagnostic tests are limited. Protective immunity against liver flukes in sheep is low or lacking, and vaccines are not yet available. Treatment and control possibilities are challenging, and resistance to flukicide drugs is increasing. In addition, climate change with warmer and more humid weather will have a substantial effect on the establishment of both flukes and snails and will most likely increase the future distribution of F. hepatica. [ABSTRACT FROM AUTHOR]

Background: MiRNAs are small non-coding RNAs that post-transcriptionally regulate gene expression in organisms ranging from viruses to mammals. There is great relevance in understanding how miRNAs regulate genes involved in the growth, development, and maturation of the many parasitic worms (helminths) that together afflict more than 2 billion people. Results: Here, we describe the miRNAs expressed by each of the predominant intra-mammalian development stages of Fasciola hepatica, a foodborne flatworm that infects a wide range of mammals worldwide, most importantly humans and their livestock. A total of 124 miRNAs were profiled, 72 of which had been previously reported and three of which were conserved miRNA sequences described here for the first time. The remaining 49 miRNAs were novel sequences of which, 31 were conserved with F. gigantica and the remaining 18 were specific to F. hepatica. The newly excysted juveniles express 22 unique miRNAs while the immature liver and mature bile duct stages each express 16 unique miRNAs. We discovered several sequence variant miRNAs (IsomiRs) as well as miRNA clusters that exhibit strict temporal expression paralleling parasite development. Target analysis revealed the close association between miRNA expression and stage-specific changes in the transcriptome; for example, we identified specific miRNAs that target parasite proteases known to be essential for intestinal wall penetration (cathepsin L3). Moreover, we demonstrate that miRNAs fine-tune the expression of genes involved in the metabolic pathways that allow the parasites to move from an aerobic external environment to the anerobic environment of the host. Conclusions: These results provide novel insight into the regulation of helminth parasite development and identifies new genes and miRNAs for therapeutic development to limit the virulence and pathogenesis caused by F. hepatica. [ABSTRACT FROM AUTHOR]

The identification of extracellular vesicles (EVs) in Fasciola hepatica has provided a new way to understand parasite-host communication. Most of the studies on EVs have focused on the adult stage of F. hepatica , but recently, the presence of EVs from different developmental stages has been reported. To better understand the potential role of EVs in the biology of the parasite and in the infection process, the protein cargo of EVs from embryonated eggs and newly-excysted juvenile (NEJs) flukes cultured up to 28 days, has been analyzed. EVs were isolated by size exclusion chromatography and evaluated by nanoparticle tracking analysis and transmission electron microscopy. LC-MS/MS proteomic analysis of EVs revealed the presence of 23 different proteins from embryonated egg-derived EVs and 29 different proteins from NEJ-derived EVs. Most of the identified proteins had been previously described in EVs from F. hepatica adults, including cytoskeletal proteins, glycolytic enzymes, stress-related proteins and tetraspanins. Nevertheless, EVs from hatching eggs and NEJs exhibited qualitative differences in composition, when compared to EVs form adults, including the absence of cathepsin cysteine peptidases. The differential content of the EVs released by the different developmental stages of the parasite reflect the intense activity of NEJs at this early stage, with several proteins involved in membrane traffic and cell physiology. This new set of identified proteins could help to understand key metabolic, biochemical and molecular mechanisms mediated by EVs that take place upon egg hatching and after parasite excystment. [ABSTRACT FROM AUTHOR]

Fasciola hepatica and Fasciola gigantica are two trematodes (liver flukes) that cause water- and foodborne disease Fasciolosis. Many animals are involved in the life cycle of the Fasciola spp., such as sheep, cattle, and camels, and humans are the definitive host. This work investigated the prevalence of Fasciola among imported and local slaughtered sheep and the relation between infection rate and seasons of the year. A total of 720,290 local and imported sheep (251,701 and 468,489, respectively) were slaughtered in a Jeddah slaughterhouse (Western region of Saudi Arabia) over the periods of 2017-2018, 2018-2019, and 2019-2020. In the three years of inspection, the infection rate at slaughter was higher among imported than local sheep. In 2017-2018, both local and imported sheep had the highest level of infection in the spring and the lowest in the winter. However, in 2018-2019, the highest rate of infection for both was documented in the summer and the lowest in the winter. Lastly, in 2019-2020, the highest rate of infection for local sheep was observed in the summer of 2020 and the lowest rate of infection was observed in the spring of 2020. Among imported sheep, the highest rate of infection was in the summer and the lowest was in winter for both 2019 and 2020. To control this disease, there is the need to apply for successful integrated programs and practice adaptive management techniques. [ABSTRACT FROM AUTHOR]

Buffaloes, as highly susceptible definitive hosts of Fasciola gigantica, suffer from a high infection rate of fasciolosis, which causes enormous economic losses. Repeat infection is responsible for this high rate; thus, elucidating the protective immunity mechanism in repeat infection is decisive in fasciolosis prevention. Herein, a secondary experimental infection model was established to preliminarily reveal the protective immunity that occurs in repeat infection. In brief, animals were assigned to three groups: group A (uninfected control), group B (primary infection) and group C (secondary infection). Buffaloes were autopsied 20 weeks post-infection for measurements of the recovered flukes and hepatic examination. In addition, the detection of specific antibody (IgG) responses to F. gigantica excretory-secretory product (FgESP) throughout the whole period and weight gain throughout the first 4 months as a percentage (%) of the starting weight were also determined. The serum hepatic enzyme gamma glutathione transferase (GGT) levels were monitored to assess hepatic damage throughout the study period. Infection establishment was compared between group B and group C. Similar specific IgG patterns were observed between group B and group C, and hepatic damage was more severe in group C than group B. Significant differences in weight gain as a percentage of the start weight were observed between group A and group B at the 3rd and 4th months postprimary infection, while significant differences were not observed between group A and group C or group B and group C. Our results suggest that challenge infection cannot induce resistance against F. gigantica in buffaloes, which is consistent with the protective immunity against Fasciola hepatica reinfection observed in sheep and goats. [ABSTRACT FROM AUTHOR]

In the present study, 34 and 33 haplotypes were detected from 144 sequences of I cox1 i and I nad1 i genes, respectively, and the Hami population had the highest number of haplotypes and the highest and K values. The present study has for the first time revealed the molecular characteristics, population genetic relationships and dispersal patterns of I F. hepatica i in sheep in Xinjiang, China, which is of great importance for the better understanding of its genetic diversity and the developing of reasonable management measures to control sheep fascioliasis. Keywords: F. hepatica; genetic diversity; population structure EN F. hepatica genetic diversity population structure 199 207 9 07/06/22 20220601 NES 220601 Introduction Fascioliasis is a parasitic disease caused by the I Fasciola hepatica i and I Fasciola gigantica i trematodes and their hybrids. Genetic diversity analysis of the three populations taking 144 sequences into consideration identified 34 and 33 haplotypes based on the I cox1 i and I nad1 i genes, respectively (Table 1). [Extracted from the article]

Triclabendazole (TCBZ) resistance is an emerging problem in fascioliasis that is not well understood. Studies including small numbers of parasites fail to capture the complexity of susceptibility variations between and within Fasciolahepatica populations. As the first step to studying the complex resistant phenotype–genotype associations, we characterized a large sample of adult F. hepatica with diverging TCBZ susceptibility. We collected parasites from naturally infected livestock slaughtered in the Cusco and Cajamarca regions of Peru. These parasites were exposed to TCBZ sulfoxide (TCBZ.SO) in vitro to determine their susceptibility. We used a motility score to determine the parasite's viability. We titrated drug concentrations and times to detect 20% non-viable (susceptible conditions) or 80% non-viable (resistant conditions) parasites. We exposed 3348 fully motile parasites to susceptible (n = 1565) or resistant (n = 1783) conditions. Three hundred and forty-one (21.8%) were classified as susceptible and 462 (25.9%) were classified as resistant. More resistant parasites were found in Cusco than in Cajamarca (p < 0.001). Resistant parasites varied by slaughterhouse (p < 0.001), month of the year (p = 0.008), fluke length (p = 0.016), and year of collection (p < 0.001). The in vitro susceptibility to TCBZ.SO in wildtype F. hepatica was associated with geography, season, and morphometry. [ABSTRACT FROM AUTHOR]

The liver fluke, Fasciola hepatica, is an obligate blood-feeder, and the gastrodermal cells of the parasite form the interface with the host's blood. Despite their importance in the host–parasite interaction, in-depth proteomic analysis of the gastrodermal cells is lacking. Here, we used laser microdissection of F. hepatica tissue sections to generate unique and biologically exclusive tissue fractions of the gastrodermal cells and tegument for analysis by mass spectrometry. A total of 226 gastrodermal cell proteins were identified, with proteases that degrade haemoglobin being the most abundant. Other detected proteins included those such as proton pumps and anticoagulants which maintain a microenvironment that facilitates digestion. By comparing the gastrodermal cell proteome and the 102 proteins identified in the laser microdissected tegument with previously published tegument proteomic datasets, we showed that one-quarter of proteins (removed by freeze–thaw extraction) or one-third of proteins (removed by detergent extraction) previously identified as tegumental were instead derived from the gastrodermal cells. Comparative analysis of the laser microdissected gastrodermal cells, tegument, and F. hepatica secretome revealed that the gastrodermal cells are the principal source of secreted proteins, as well as showed that both the gastrodermal cells and the tegument are likely to release subpopulations of extracellular vesicles (EVs). Microscopical examination of the gut caeca from flukes fixed immediately after their removal from the host bile ducts showed that selected gastrodermal cells underwent a progressive thinning of the apical plasma membrane which ruptured to release secretory vesicles en masse into the gut lumen. Our findings suggest that gut-derived EVs are released via a novel atypical secretory route and highlight the importance of the gastrodermal cells in nutrient acquisition and possible immunomodulation by the parasite. [ABSTRACT FROM AUTHOR]

Fascioliasis, which is caused by infection with Fasciola gigantica and Fasciola hepatica, is a zoonotic disease with a global distribution. This comparative study aimed to investigate antioxidant enzyme activities and oxidative status of chronic fascioliasis patients. In this study, 20 patients were compared with 10 controls and the levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPX), catalase (CAT), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes were evaluated. The results showed that MDA, CAT, AST, and ALT levels were higher in patients than in controls, while SOD and GPX levels were higher in controls than in patients (P≤0.05). Moreover, the enzyme MDA showed a positive correlation with SOD and GPX in the infected group. The enzyme SOD had an indirect correlation with CAT and a direct correlation with GPX. The positive correlation between ALT and AST was shown to be extremely significant (P≤0.05). The significant decrease in antioxidant enzymes and an increase in serum lipid peroxidation in the red blood cells of patients with fascioliasis indicated the presence of oxidative stress, which showed inflammation and oxidative stress, the pathogenesis of which was indicative of the stage of infection. [ABSTRACT FROM AUTHOR]

Type 1 diabetes (T1D) is an autoimmune disease caused by the destruction of the insulin‐producing β‐cells within the pancreas. Islet transplantation represents one cure; however, during islet preparation and post transplantation significant amounts of β‐cell death occur. Therefore, prevention and cure of T1D is dependent upon the preservation of β‐cell function and the prevention of β‐cell death. Phosphoinositide 3‐kinase (PI3K)/Akt signaling represents a promising therapeutic target for T1D due to its pronounced effects on cellular survival, proliferation, and metabolism. A growing amount of evidence indicates that PI3K/Akt signaling is a critical determinant of β‐cell mass and function. Modulation of the PI3K/Akt pathway, directly (via the use of highly specific protein and peptide‐based biologics, excretory/secretory products of parasitic worms, and complex constituents of plant extracts) or indirectly (through microRNA interactions) can regulate the β‐cell processes to ultimately determine the fate of β‐cell mass. An important consideration is the identification of the specific PI3K/Akt pathway modulators that enhance β‐cell function and prevent β‐cell death without inducing excessive β‐cell proliferation, which may carry carcinogenic side effects. Among potential PI3K/Akt pathway agonists, we have identified a novel parasite‐derived protein, termed FhHDM‐1 (Fasciola hepatica helminth defense molecule 1), which efficiently stimulates the PI3K/Akt pathway in β‐cells to enhance function and prevent death without concomitantly inducing proliferation unlike several other identified stimulators of PI3K/Akt signaling. As such, FhHDM‐1 will inform the design of biologics aimed at targeting the PI3K/Akt pathway to prevent/ameliorate not only T1D but also T2D, which is now widely recognized as an inflammatory disease characterized by β‐cell dysfunction and death. This review will explore the modulation of the PI3K/Akt signaling pathway as a novel strategy to enhance β‐cell function and survival. [ABSTRACT FROM AUTHOR]

Fascioliasis caused by Fasciola species (Fasciola hepatica and Fasciola gigantica) is considered as the most important helminthic infection of domestic animals in developing countries. The present study was performed during January to December 2019, in Biology Department, University of Zakho. Hundred adult's flukes of Fasciola spp. were isolated from bile ducts and gallbladder of cattle, sheep and goats slaughtered at different abattoirs (Duhok, Zakho, Shelidiza, Aqrah and Bardarash) in Duhok governorate, Kurdistan region, Iraq. Genomic DNA extraction was performed using a Genomic DNA Extraction kit, Jena Bioscience GmbH (Germany). Specific markers ITS1 and ITS2 ribosomal DNA were used for amplifying 480 and 550 bps fragments. DNA sequences have been used to characterize these liver flukes. The PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with RedSafe dye, and photographed. ITS-1 and ITS-1 markers were amplified successfully, the length of produced band for ITS-1 was 480 bp and for ITS-2 was 550 bp. Thirteen Fasciola spp. flukes isolated from cattle, sheep and goats from different districts were sequenced, from these thirteen PCR products, 7 amplicons were recognized as Fasciola hepatica and 6 amplicons as Fasciola gigantica. These sequences were deposited in GenBank under accession numbers: MW161261, MW084365, MW052601 MW084349, MW082585, MW084348, MW082830 for F. hepatica and MW161260, MW052602, MW084353, MW084350, MW082589, MW085034 for F. gigantica. This study confirms the prevalence of Fasciola hepatica and Fasciola gigantica in Iraq by using the ITS-1 and ITS-2 rDNA. [ABSTRACT FROM AUTHOR]

Understanding drug penetration, distribution, and metabolization is fundamental for understanding drug efficacy. This also accounts for parasites during antiparasitic treatment. Recently, we established matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) in blood flukes and liver flukes. This label-free technique is capable of visualizing the molecular distribution of endogenous and exogenous molecules, such as drug compounds. Here, we conducted atmospheric-pressure scanning microprobe MALDI MSI (AP-SMALDI MSI) of tissue sections of adult Fasciola hepatica that have been treated in vitro with 100 µM of triclabendazole (TCBZ), the drug of choice for treatment of fasciolosis, and its main metabolite triclabendazole sulfoxide (TCBZ-SO). Measurements covered an m/z mass range of 250–1,000 and provided a high spatial resolution using a pixel size of 10 µm. To support the interpretation of drug distribution, we first identified endogenous lipids that mark characteristic tissues such as the gastrodermis, the tegument, and the parenchyma. The obtained results suggested an early tegumental route of TCBZ uptake within 20 min, followed by spreading throughout the parasite after 4 h, and an even distribution in most tissues after 12 h. This coincided with a strong reduction of parasite vitality. TCBZ-SO treatment demonstrated the accumulation of this metabolite in the same tissues as the parent drug compound. These data demonstrate the auspicious potential of MALDI MSI to visualize uptake and distribution patterns of drugs or drug-candidate compounds in parasites, which might contribute to preclinical drug discovery in liver fluke research and beyond. [ABSTRACT FROM AUTHOR]

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-a, and promoted the expression of transforming growth factor-ß and interleukin-10. These results showed that Fh- Cystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection. [ABSTRACT FROM AUTHOR]

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