Liver Fluke Infection Research Articles

Spatial prediction of the probability of liver fluke infection in water resource within sub-basin using an optimized geographically-weighted regression model

Spatial prediction of the probability of liver fluke infection in water resource within sub-basin using an optimized geographically-weighted regression model

Molecular characterization of Fasciola hepatica obtained from cattle and horse in Central Chile

Liver fluke infection, caused by the trematode Fasciola hepatica, is a parasitic zoonotic disease affecting various mammals, including humans, and has significant implications for public, animal, and ecosystem health. This study provides the first genetic characterization of F. hepatica in Chile, focusing on the complete mitochondrial gene cox1. Samples were collected from two different host species: cattle and horses. Our findings revealed that 70 % of detected haplotypes were found in either cattle or horses, which coincides with their geographical origin. Interestingly, the use of full-length sequences resulted in the identification of 80 % unique sequences, whereas this reduced to 45 % when analyzing the traditionally used short sequences. This underestimation of genetic diversity suggests that broader sequencing efforts might be essential for a more accurate understanding of F. hepatica genetic landscape. This research underscores the importance of understanding the genetic variability in parasites to improve strategies for disease control and treatment.

A Comprehensive Multi-Omics Study of Serum Alterations in Red Deer Infected by the Liver Fluke Fascioloides magna

Liver fluke infections are acknowledged as diseases with global prevalence and significant implications for both veterinary and public health. The large American liver fluke, Fascioloides magna, is a significant non-native parasite introduced to Europe, threatening the survival of local wildlife populations. The aim of this study was to analyze differences in the serum proteome and metabolome between F. magna-infected and control red deer. Serum samples from red deer were collected immediately following regular hunting operations, including 10 samples with confirmed F. magna infection and 10 samples from healthy red deer. A proteomics analysis of the serum samples was performed using a tandem mass tag (TMT)-based quantitative approach, and a metabolomics analysis of the serum was performed using an untargeted mass spectrometry-based metabolomics approach. A knowledge-driven approach was applied to integrate omics data. Our findings demonstrated that infection with liver fluke was associated with changes in amino acid metabolism, energy metabolism, lipid metabolism, inflammatory host response, and related biochemical pathways. This study offers a comprehensive overview of the serum proteome and metabolome in response to F. magna infection in red deer, unveiling new potential targets for future research. The identification of proteins, metabolites, and related biological pathways enhances our understanding of host–parasite interactions and may improve current tools for more effective liver fluke control.

Risk factors of cholangiocarcinoma in areas not endemic for liver fluke infection.

Thailand has the world's highest prevalence of cholangiocarcinoma (CCA), especially in the endemic area of liver fluke Opisthorchis viverrini infection. However, other regions of Thailand still have relatively high CCA prevalence. We aimed to determine CCA risk factors in areas not endemic for OV infection. A case--control study was performed at a referral center during December 2016-December 2017. We collected blood samples and information from CCA patients and identified them as cases. The control group comprised patients who visited a gastrointestinal clinic for colorectal cancer screening colonoscopy. Logistic regression analysis was used to determine risk factors for CCA. Of 138 participants, O. viverrini infection rate was higher in the case than in the control group (57.1% vs. 36.1%, P = 0.023). Male, O. viverrini infection, smoking, alcohol consumption, and biliary tract diseases were independent risk factors, whereas diabetes, obesity, and cirrhosis were not associated with CCA. By age and sex-adjusted analysis, chronic biliary tract diseases, especially choledochal cysts and smoking, were risk factors for CCA, with adjusted odds ratio (aOR) of 12.7 (95% confidence interval [CI]: 1.4-116.9) and 3.8 (95% CI: 1.3-11.8), respectively, while O. viverrini infection became insignificant risk for CCA (aOR 1.8, 95% CI: 0.8-4.1). In contrast with endemic areas for O. viverrini infection, chronic biliary tract diseases and smoking are major risk factors, whereas O. viverrini infection has trivial contribution to the development of CCA.

Detection of the carcinogenic liver fluke, Opisthorchis viverrini: comparison of two coprological methods versus the automatic feces analyzer.

Liver fluke infection, particularly Opisthorchis viverrini, poses a significant public health risk in Thailand, where it is closely associated with cholangiocarcinoma and contributes to substantial mortality in the northeastern region. Diagnosis of this condition employs various parasitological approaches. This research aims to compare the diagnostic accuracy of three parasitological techniques: the Kato Katz technique (KKT), the formalin-ethyl acetate concentration technique (FECT), and the Fully Automatic Feces Analyzer (FAFA) for O. viverrini identification. A total of 455 fecal specimens were collected from rural areas across five provinces in northeastern Thailand. The specimens were processed according to each method and examined through microscopy for KKT and FECT, and by utilizing an artificial intelligence-based machine for FAFA. Data analysis was conducted to assess parasitic infection rates and observe diagnostic accuracy. The results revealed a parasitic infection rate of 19.34%, with the majority of infections attributed to O. viverrini (18.02%), followed by Strongyloides stercoralis (0.88%). FECT exhibited the highest positive detection of O. viverrini eggs (16.48%), followed by FAFA (10.55%), and KKT (8.57%), respectively. Statistical analysis indicated sensitivity and specificity values for O. viverrini detection by KKT (100% and 89.21%), FECT (98.67% and 97.63%), and FAFA (97.92% and 91.15%). The positive predictive value, negative predictive value, and kappa were reported for FECT (89.16%, 99.73%, 0.92), FAFA (56.63%, 99.73%, 0.67), and KKT (45.78%, 100%, 0.58). Additionally, the preparation time for KKT, FECT, and FAFA was 30, 15, and 10 min, respectively. In conclusion, this study highlights FECT, KKT, and FAFA as comparably sensitive in diagnosing O. viverrini. The FAFA machine emerges as a potentially valuable tool for detecting O. viverrini and other parasitic infections, showcasing promise for clinical use. The findings provide valuable insights into the diagnostic landscape and underscore the potential of FAFA in enhancing efficiency and accuracy in parasitological assessments.

Construction and validation of a novel multi-epitope in silico vaccine design against the paramyosin protein of Opisthorchis viverrini using immunoinformatics analyses

Liver fluke infection caused by Opisthorchis viverrini (O. viverrini) remains a significant but neglected health threat across Southeastern Asia. The early infective anabolic growth stage of O. viverrini expresses and exposes proteins integral for the growth and maturation of immature worms to the adult catabolic stage. Among these proteins, paramyosin emerged as a distinct immunogenic protein during opisthorchiasis. The functional region of the paramyosin protein known as myosin tail was selected to design a multi-epitope vaccine (MEV) to elicit T and B cell immune responses in susceptible human hosts utilizing various immunoinformatics and in silico vaccinology tools. The vaccine candidate had several B- and T-cell epitopes that stimulate both humoral and cellular immune responses. Moreover, in silico structural, docking, and dynamic analyses showed that the construct interacted with target immune receptors effectively, which may result in sufficient immunological stimulation. Analysis of simulated coverage efficacy also supports vaccine application in the field. Cloning and expression of the vaccine candidate were determined to be viable based on physicochemical and in silico assessments. These results reveal that the vaccine candidate developed herein is stable and potentially useful in addressing opisthorchiasis. The promising result of this study establishes a strong platform for initiating laboratory and efficacy trials for the vaccine candidate.

Assessment of accuracy of liver fluke diagnostic tests using the gold standard of total worm counts

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 loge (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.

Update on liver fluke in sheep and cattle

Fasciola hepatica, the common liver fluke, affects the health and welfare of grazing animals worldwide. Liver fluke infection has a significant economic cost to the UK agriculture industry through sheep deaths, reduced growth rates, fertility and milk production in sheep and cattle and condemned livers at the abattoir. Changing weather patterns in the UK over recent years have often resulted in milder winters and summers that may be hot or wet but are above all unpredictable, affecting the epidemiology of liver fluke. This poses a risk for farmers relying on traditional calendar-based treatment routines, who may be caught out by liver fluke risk periods that are longer and extend beyond what would traditionally have been expected. Triclabendazole resistance is an increasing problem in many parts of the country and with no new flukicides on the horizon, existing products need to be used responsibly to ensure they remain useful for as long as possible.

Early stage Fasciola hepatica infection in growing cattle: impact on weight gain and liver weight determined in flukicide efficacy studies.

Although the negative impact of liver fluke (Fasciola hepatica) infection on production and health in cattle is generally accepted, results of individual research have been variable, ranging from important negative impacts on the animal to minimal or no impact. To add information on the impact of F. hepatica infection in growing cattle, weight gain and liver weight of young experimentally infected animals from seven controlled efficacy studies were analyzed. In each study, fluke naïve animals were inoculated with approximately 450 to 500 F. hepatica encysted metacercariae, blocked on body weight and randomly assigned into one untreated group (controls) and groups which were administered an experimental flukicide when the flukes were 4 weeks old (migrating) and sacrificed 8 weeks thereafter (12 weeks after inoculation). Data of groups which demonstrated >90% reduction of fluke counts following treatment and groups left untreated (total 103 and 47 animals, respectively) were compared. There was a significant (p < 0.0001) negative association between fluke count and weight gain while fluke count and liver weight and fluke count and relative liver weight were positively associated (p < 0.0001). Over the 8-week post-treatment period, flukicide-treated cattle had almost 15% more weight gain than the controls (50.9 kg vs. 44.4 kg; p = 0.0003). Absolute and relative liver weight was significantly (p < 0.0001) lower in flukicide-treated compared to untreated cattle. Overall, this analysis provided evidence of a substantial negative effect of early (migrating) liver fluke infection on the growth of young cattle, likely due to pathology of the liver and associated reduction in its function as the central organ for bioenergy and protein metabolism.

Quantitative proteomics analysis reveals possible anticancer mechanisms of 5'-deoxy-5'-methylthioadenosine in cholangiocarcinoma cells.

Cholangiocarcinoma (CCA) is an aggressive cancer originating from bile duct epithelium, particularly prevalent in Asian countries with liver fluke infections. Current chemotherapy for CCA often fails due to drug resistance, necessitating novel anticancer agents. This study investigates the potential of 5'-deoxy-5'-methylthioadenosine (MTA), a naturally occurring nucleoside, against CCA. While MTA has shown promise against various cancers, its effects on CCA remain unexplored. We evaluated MTA's anticancer activity in CCA cell lines and drug-resistant sub-lines, assessing cell viability, migration, invasion, and apoptosis. The potential anticancer mechanisms of MTA were explored through proteomic analysis using LC-MS/MS and bioinformatic analysis. The results show a dose-dependent reduction in CCA cell viability, with enhanced effects on cancer cells compared to normal cells. Moreover, MTA inhibits growth, induces apoptosis, and suppresses cell migration and invasion. Additionally, MTA enhanced the anticancer effects of gemcitabine on drug-resistant CCA cells. Proteomics revealed the down-regulation of multiple proteins by MTA, affecting various molecular functions, biological processes, and cellular components. Network analysis highlighted MTA's role in inhibiting proteins related to mitochondrial function and energy derivation, crucial for cell growth and survival. Additionally, MTA suppressed proteins involved in cell morphology and cytoskeleton organization, important for cancer cell motility and metastasis. Six candidate genes, including ZNF860, KLC1, GRAMD1C, MAMSTR, TANC1, and TTC13, were selected from the top 10 most down-regulated proteins identified in the proteomics results and were subsequently verified through RT-qPCR. Further, KLC1 protein suppression by MTA treatment was confirmed through Western blotting. Additionally, based on TCGA data, KLC1 mRNA was found to be upregulated in the tissue of CCA patients compared to that of normal adjacent tissues. In summary, MTA shows promising anticancer potential against CCA by inhibiting growth, inducing apoptosis, and suppressing migration and invasion, while enhancing gemcitabine's effects. Proteomic analysis elucidates possible molecular mechanisms underlying MTA's anticancer activity, laying the groundwork for future research and development of MTA as a treatment for advanced CCA.

Zoonotic human liver flukes, a type 1 biocarcinogen, in freshwater fishes: genetic analysis and confirmation of molluscan vectors and reservoir hosts in Bangladesh

BackgroundOpisthorchiid flukes, particularly Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, and Metorchis spp. are the most common fish-borne zoonotic human liver flukes (hLFs). Liver fluke infections are more prevalent in resource-deprived and underprivileged areas. We herein estimated the prevalence of the metacercariae (MC) of major hLFs in common large freshwater fishes (lFWF) marketed for human consumption from some selected areas of Bangladesh along with detection of their molluscan vectors and reservoirs.MethodsThe current status of fish-borne zoonotic hLF infections in lFWF was investigated along with their molluscan vectors and mammalian reservoir hosts in Mymensingh and Kishoreganj in Bangladesh from July 2018–June 2022 using conventional and multiple molecular techniques, such as PCR, PCR-restriction fragment length polymorphism (RFLP), sequencing, and bioinformatic analyses. The infection rate of fishes was analyzed using the Z-test and the loads of MC were compared using the chi-squared (χ2) test.ResultsThe MC of C. sinensis, Opisthorchis spp., and Metorchis spp. were detected in 11 species of common and popular lFWF. In lFWF, the estimated prevalence was 18.7% and the mean load was 137.4 ± 149.8 MC per 100 g of fish. The prevalence was the highest (P < 0.05) in spotted snakehead fishes (Channa punctata, 63.6%). The highest rate of infection (P < 0.05) was observed with the MC of C. sinensis (11.8%). Metacercariae were almost equally (P > 0.05) distributed between the head and body of fishes. The infection rate was slightly higher in cultured (19.6%) fishes. The MC of C. sinensis, O. felineus, O. viverrini, and Metorchis orientalis in fishes were confirmed using PCR, PCR-RFLP and bioinformatics. The cercariae of opisthorchiid (Pleurolophocercus cercariae) flukes were only recovered from Bithynia spp. (3.9%, 42 out of 1089). The ova of hLFs from dogs (4.3%, 5 out of 116) and cats (6.0%, 6 out of 100), and adult flukes (M. orientalis) from ducks (41.1% 113 out of 275) were detected.ConclusionsThe MC of hLFs are highly prevalent in fresh water fishes in Bangladesh. Reservoir hosts, such as street dogs, cats, and ducks carried the patent infection, and residents of Bangladesh are at risk.Graphical

The landscape of etiological patterns of hepatocellular carcinoma and intrahepatic cholangiocarcinoma in Thailand.

Thailand is among countries with the highest global incidence and mortality rates of hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (iCCA). While viral hepatitis and liver fluke infections have been associated with HCC and iCCA, respectively, other environmental risk factors, overall risk factor commonality and combinatorial roles, and effects on survival have not been systematically examined. We conducted a TIGER-LC consortium-based population study covering all high-incidence areas of both malignancies across Thailand: 837 HCC, 1474 iCCA, and 1112 controls (2011-2019) were comprehensively queried on lifelong environmental exposures, lifestyle, and medical history. Multivariate logistic regression and Cox proportional hazards analyses were used to evaluate risk factors and associated survival patterns. Our models identified shared risk factors between HCC and iCCA, such as viral hepatitis infection, liver fluke infection, and diabetes, including novel and shared associations of agricultural pesticide exposure (OR range of 1.50; 95% CI: 1.06-2.11 to 2.91; 95% CI: 1.82-4.63) along with vulnerable sources of drinking water. Most patients had multiple risk factors, magnifying their risk considerably. Patients with lower risk levels had better survival in both HCC (HR 0.78; 95% CI: 0.64-0.96) and iCCA (HR 0.84; 95% CI: 0.70-0.99). Risk factor co-exposures and their common associations with HCC and iCCA in Thailand emphasize the importance for future prevention and control measures, especially in its large agricultural sector. The observed mortality patterns suggest ways to stratify patients for anticipated survivorship and develop plans to support medical care of longer-term survivors, including behavioral changes to reduce exposures.

Benzimidazole‐associated pancytopenia in a dog with presumed Heterobilharzia americana infection

AbstractAdverse drug events are classified into type A (dose‐dependent and common) or type B (idiosyncratic and uncommon) reactions. Benzimidazole toxicosis in veterinary medicine typically induces nonspecific signs such as gastrointestinal distress and infrequent myelosuppression. Pancytopenia from benzimidazoles is exceptionally rare, with only one reported case. We present a 12‐year‐old, female, spayed, mixed‐breed dog with acute lethargy, anorexia, pyrexia and severe leukopenia after high‐dose benzimidazole therapy for suspected liver fluke infection. Despite hospitalisation, the dog remained unresponsive to antimicrobial therapies, developed neutropenic sepsis and was euthanized. Bone marrow analysis indicated marked haematopoietic hypoplasia suggestive of a type A adverse drug event secondary to an incidental, high‐dose fenbendazole administration.

Estimation of diagnostic sensitivity and specificity of abattoir registrations and bulk tank milk ELISA as herd-level tests for Fasciola hepatica using Bayesian latent class modelling

The common liver fluke, Fasciola hepatica, is a trematode parasite found worldwide, typically with a focal distribution due to its requirement for suitable climatic and environmental conditions to complete its lifecycle. Bovine fasciolosis causes suboptimal production and economic losses, including liver condemnation at slaughter. The lack of reliable diagnostic methods is a disadvantage to the increasing demand for surveillance and control. The aim of this study was to evaluate the diagnostic accuracy of bulk tank milk (BTM) antibody testing and aggregated abattoir registrations (AAR) of liver fluke as herd-level tests for F. hepatica infection using Bayesian latent class models. Data from the abattoirs in 2019–2021 and BTM, sampled in the winter of 2020/2021, were collected from 437 herds on the southwest coast of Norway. The BTM samples were analysed with the SVANOVIR® F. hepatica-Ab ELISA test, with results given as an optical density ratio (ODR), and later dichotomized using the recommended cut-off value from the test manufacturer (ODR ≥0.3). Based on the BTM ELISA test, 47.8% of the herds tested positive. The AAR test was defined as the herd-level proportion of female slaughtered animals registered with liver fluke infection during the study period. For this test, three cut-offs were used (a proportion of 0.05, 0.1 and 0.2). The herds were split into two subpopulations (“Coastal” and “Inland”), which were expected to differ in true prevalence of F. hepatica infection based on climate-related and geographical factors. The diagnostic accuracies of both tests were estimated using Bayesian latent class models with minimally informative priors. Post-hoc analysis revealed that the maximum sum of sensitivity (Se) and specificity (Sp) of the tests was achieved with a herd-level proportion of ≥0.1 registered with liver fluke as the AAR test. Using this cut-off, the median estimate for the diagnostic accuracy of the BTM ELISA was 90.4% (84.0–96.2 95% Posterior Credible Interval (PCI)) for Se and 95.3% (90.6–100% PCI) for Sp, while the median estimate of Se for AAR was 87.5% (81.4–93.1% PCI) and the median estimate of Sp for AAR was 91.0% (85.2–96.5% PCI). The cut-off evaluation of the SVANOVIR® F. hepatica-Ab ELISA test for BTM confirmed the manufacturer's recommended cut-off of ODR ≥0.3 to denote positive and negative herds. This study suggests that AAR and BTM ELISA test can be used as herd-level tools to monitor liver fluke infection, so that appropriate interventions against infection can be implemented as necessary.

Correlation between the Fasciola hepatica infection rate (number of parasites in the liver parenchyma) and the antibody titration, detected by ELISA assay.

This study aimed to demonstrate the existence of a possible correlation between the level of liver fluke infection in cattle caused by Fasciola hepatica and the titers of anti-parasite antibodies in the blood. 113 blood samples were taken from non-dewormed pasture cattle in the communal slaughterhouse of the Jijel city in order to determine the serological titration of anti-fluke antibodies by the ELISA method. After slaughtering the animals studied, a count of parasites present in the inspected livers was carried out in order to assess the level of parasitic infection. The prevalence of bovine fasciolosis (due to Fasciola hepatica) based on counting the number of flukes in the inspected livers was around 61.06%, while the seroprevalence obtained is 55.75%. The sensitivity and specificity of the ELISA test used in this study were, respectively, 91.3% and 100%. A very strong correlation (r = 0.93 to p < 0.0001) was statistically demonstrated between the antibody titers and the number of flukes in the inspected livers. According to the results of this study, there is a strong statistical correlation between serum antibody titers and the number of flukes present in the liver (the higher the number of parasites in the liver, the higher the level of anti-flukes antibodies in the blood).

Identification and Molecular Characterization of Giant Liver Fluke (Fascioloides magna) Infection in European Fallow Deer (Dama dama) in Romania-First Report.

Fascioloidosis is a parasitic disease of primary wild and domestic ruminants, caused by giant liver fluke, Fascioloides magna. The definitive host of the liver fluke in its area of origin (North America) is the white-tailed deer (Odocoileus virginianus). In Europe, the red deer (Cervus elaphus) and European fallow deer (Dama dama) are definitive hosts and the most sensitive hosts to F. magna infection, on which the parasite exerts serious pathogenic effects. In this study, we analyzed fecal samples and livers of 72 D. dama from 11 hunting grounds in Arad County, Romania. Of the 72 fecal samples and livers from D. dama, trematodes of the genus Fascioloides were identified in four (5.56%). Sequencing revealed that the trematodes identified in the samples were similar to the sequence of F. magna (GenBank no. EF534992.1, DQ683545.1, KU232369.1). The sequence obtained from the molecular analysis has been deposited in GenBank® under accession number OQ689976.1. This study describes the first report of giant liver fluke (F. magna) infection in D. dama in Romania.

Clonorchis sinensis: a mimic of hepatic mucinous cystic neoplasm or just a fluke?

Liver flukes, endemic in East Asia, are parasites that can infect humans and cause liver and bile duct disease. While most infected individuals are asymptomatic, chronic infection can lead to structural hepatobiliary manifestations including hepatomegaly, intrahepatic bile duct dilatation, epithelial hyperplasia, periductal fibrosis, and potentially cholangiocarcinoma. There are no reports of human cases of liver fluke infection presenting as a hepatic cystic lesion. We present the case of a 52-year-old asymptomatic Chinese immigrant presenting with a suspected mucinous cystic neoplasm of the liver. CT and Ultrasound examinations demonstrated an enlarging complex cyst involving segments II and III. Liver function tests, hydatid serology and tumour markers were normal. He underwent elective laparoscopic left hemi-hepatectomy with liver fluke discovered on histology. Subsequent genetic testing confirmed Clonorchis sinensis infection. The patient made an uneventful recovery from surgery and was treated with anthelminthic therapy. This case highlights the importance of considering liver fluke as a differential diagnosis for hepatic cystic lesions in patients from endemic regions. Appropriate diagnosis could avoid surgery, whilst targeted anthelminthic therapy minimises the risk of chronic infection and associated complications, including cholangiocarcinoma.

Plastoquinone-Derivative SkQ1 Improved the Biliary Intraepithelial Neoplasia during Liver Fluke Infection.

Carcinogenic food-borne liver fluke infections are a serious epidemiological threat worldwide. The major complications of Opisthorchis felineus infection are chronic inflammation and biliary intraepithelial neoplasia. Although evidence has accumulated that increased reactive oxygen species production is observed in liver fluke infection, a direct relationship between the oxidative stress and biliary intraepithelial neoplasia has not been shown. Quinones and SkQ1, a derivative of plastoquinone, have been demonstrated to be cytoprotective in numerous liver injuries due to their potent antioxidant properties. This study is aimed to assess the level of biliary intraepithelial neoplasia in O. felineus-infected hamsters after treatment with mitochondria-targeted SkQ1. SkQ1 significantly reduced the biliary intraepithelial neoplasia, which was accompanied by a decrease in lipid and DNA oxidation byproducts, mRNA expression and level of proteins associated with inflammation (TNF-α, CD68) and fibrogenesis (CK7, αSMA), and was also associated with an activation of the Keap1-Nrf2 pathway. Thus, a direct relationship was found between oxidative stress and the severity of biliary intraepithelial neoplasia in O. felineus-infected hamsters. The hepatoprotective effect of plastoquinone-derivative SkQ1 was established; therefore, this compound is a promising agent in complex therapy in the treatment of opisthorchiasis.

Liver Flukes: Clonorchis and Opisthorchis.

Clonorchis sinensis, Opisthorchis viverrini and O. felineus are liver flukes of human and animal pathogens occurring across much of Europe and Asia. Nevertheless, they are often underestimated compared to other, better known neglected diseases in spite of the fact that many millions of people are infected and hundreds of millions are at risk. This is possibly because of the chronic nature of the infection and disease and that it takes several decades prior to a life-threatening pathology to develop. Several studies in the past decade have provided more information on the molecular biology of the liver flukes which clearly lead to better understanding of parasite biology, systematics and population genetics. Clonorchiasis and opisthorchiasis are characterized by a chronic infection that induces hepatobiliary inflammation, especially periductal fibrosis, which can be detected by ultrasonography. These chronic inflammations eventually lead to cholangiocarcinoma (CCA), a usually fatal bile duct cancer that develops in some infected individuals. In Thailand alone, opisthorchiasis-associated CCA kills up to 20,000 people every year and is therefore of substantial public health importance. Its socioeconomic impacts on impoverished families and communities are considerable. To reduce hepatobiliary morbidity and CCA, the primary intervention measures focus on control and elimination of the liver fluke. Accurate diagnosis of liver fluke infections in both human and other mammalian, snail and fish intermediate hosts is important for achieving these goals. While the short-term goal of liver fluke control can be achieved by praziquantel chemotherapy, a comprehensive health education package targeting school children is believed to be more beneficial for a long-term goal/solution. It is recommended that transdisciplinary research or multisectoral control approach including one health and/or eco health intervention strategy should be applied to combat the liver flukes and hence contribute to reduction of CCA in endemic areas.

Machine-Learning-Based Forest Classification and Regression (FCR) for Spatial Prediction of Liver Fluke Opisthorchis viverrini (OV) Infection in Small Sub-Watersheds

Infection of liver flukes (Opisthorchis viverrini) is partly due to their suitability for habitats in sub-basin areas, which causes the intermediate host to remain in the watershed system in all seasons. The spatial monitoring of fluke at the small basin scale is important because this can enable analysis at the level of the factors involved that influence infections. A spatial mathematical model was weighted by the nine spatial factors X1 (index of land-use types), X2 (index of soil drainage properties), X3 (distance index from the road network, X4 (distance index from surface water resources), X5 (distance index from the flow accumulation lines), X6 (index of average surface temperature), X7 (average surface moisture index), X8 (average normalized difference vegetation index), and X9 (average soil-adjusted vegetation index) by dividing the analysis into two steps: (1) the sub-basin boundary level was analyzed with an ordinary least square (OLS) model used to select the spatial criteria of liver flukes aimed at analyzing the factors related to human liver fluke infection according to sub-watersheds, and (2) we used the infection risk positional analysis level through machine-learning-based forest classification and regression (FCR) to display the predictive results of infection risk locations along stream lines. The analysis results show four prototype models that import different independent variable factors. The results show that Model 1 and Model 2 gave the most AUC (0.964), and the variables that influenced infection risk the most were the distance to stream lines and the distance to water bodies; the NDMI and NDVI factors rarely affected the accuracy. This FCR machine-learning application approach can be applied to the analysis of infection risk areas at the sub-basin level, but independent variables must be screened with a preliminary mathematical model weighted to the spatial units in order to obtain the most accurate predictions.