Liver Fluke Research Articles

Spatial Predictive Modeling of Liver Fluke Opisthorchis viverrine (OV) Infection under the Mathematical Models in Hexagonal Symmetrical Shapes Using Machine Learning-Based Forest Classification Regression

Infection with liver flukes (Opisthorchis viverrini) is partly due to their ability to thrive in habitats in sub-basin areas, causing the intermediate host to remain in the watershed system throughout the year. Spatial modeling is used to predict water source infections, which involves designing appropriate area units with hexagonal grids. This allows for the creation of a set of independent variables, which are then covered using machine learning techniques such as forest-based classification regression methods. The independent variable set was obtained from the local public health agency and used to establish a relationship with a mathematical model. The ordinary least (OLS) model approach was used to screen the variables, and the most consistent set was selected to create a new set of variables using the principal of component analysis (PCA) method. The results showed that the forest classification and regression (FCR) model was able to accurately predict the infection rates, with the PCA factor yielding a reliability value of 0.915. This was followed by values of 0.794, 0.741, and 0.632, respectively. This article provides detailed information on the factors related to water body infection, including the length and density of water flow lines in hexagonal form, and traces the depth of each process.

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.

MicroRNA profiling of exosomes derived from plasma and their potential as biomarkers for Opisthorchis viverrini-associated cholangiocarcinoma

Cholangiocarcinoma (CCA) is a life-threatening disease that impacts patients worldwide. In Southeast Asian countries, the liver fluke Opisthorchis viverrini plays a major role in inducing carcinogenesis of the bile ducts. Due to its asymptomatic nature, O. viverrini infections are rarely treated, consequently leading to the development of advanced stages of CCA before diagnosis. Despite the current use of exosomal microRNAs (miRNA) as diagnostic biomarkers for the early detection of many types of cancer, the applications for miRNA remain limited with CCA. Circulating exosomes, membranous vesicles essential for intercellular communication, were found to contain unique miRNA. In this study, we conducted next-generation sequencing (Ion Torrent PGM) and bioinformatics to characterize and compare the contents of exosomal miRNA derived from the plasma of CCA patients, O. viverrini-infected patients, and healthy individuals, as well as to identify and validate key molecules as markers for screening the diagnosis of CCA and O. viverrini infection. The obtained results showed the success of using NGS technology in discovering exosomal miRNAs, specifically miR-194–5p and miR-192–5p, both of which were upregulated in the O. viverrini-infected group. Interestingly, miR-192–5p was upregulated while miR-194–5p was downregulated in CCA, suggesting their potential use as biomarkers for screening CCA and O. viverrini infection, especially in O. viverrini-endemic areas.

Social Impact Assessment Project on Cholangiocarcinoma Screening and Care Program

The research aims to assess users utility and cost-effectiveness based on the results of rapid antigen test kits for urine and abdominal ultrasound methods. This study uses qualitative methods to analyze liver fluke screening among individuals aged 40 and above in four provinces. Purposive sampling was used, with 373 participants and 379 participants, respectively. Utility measurement tools included a direct Visual Analog Scale (VAS) and indirect EuroQoL (5Q-5D-5L), and willingness to pay value was measured using bidding. The result shows that the sample group has an average VAS score of 76.90, with slight pain or discomfort reported in the five health dimensions. The utility value reflecting daily quality of life is 0.89. The average willingness to pay per use for rapid test kits and abdominal ultrasounds is 187 baht and 796 baht, respectively. Reasons for not paying higher include income factors, high prices, and personal limitations. However, participants believe public hospitals could support examination costs and provide access to benefits. The project benefits people economically, socially, and culturally, and if the Ministry of Public Health and related agencies can effectively use it, it will maximize benefits.

Investigation of a Serine Protease Inhibitor Active in the Infectious Stage of the Human Liver Fluke Opisthorchis viverrini.

Serine protease inhibitors (serpins) participate in the regulation of inflammation, blood coagulation, and complement activation in humans. This research aimed to identify and characterize such inhibitors of the human liver fluke Opisthorchis viverrini. Parasite proteins that might contribute to the modulation of host physiology are of particular interest, especially as chronic opisthorchiasis increases the risk of developing biliary cancer. BLAST was used to find hypothetical serpins predicted from the parasite genome data. RNA extraction and reverse transcriptase PCR were used to isolate a serpin cDNA and to determine developmental transcript abundance. The evolutionary relation to other trematode serpins was revealed by phylogenetic analysis. Recombinant serpin was expressed in Escherichia coli and used to test the immunoreactivity of human opisthorchiasis sera and the inhibition of human serine proteases. A substantial serpin family with high sequence divergence among the members was found in the genus Opisthorchis. A serpin, different from previously analyzed trematode serpins, was cloned. The transcript was only detected in metacercariae and newly excysted juveniles. Human opisthorchiasis sera showed statistically significant reactivity to recombinant serpin. The serpin caused moderate inhibition of thrombin and low inhibition of kallikrein and chymotrypsin. This parasite serpin could be further evaluated as a diagnostic tool for early infection. Kallikrein and thrombin are involved in fibrinolysis; therefore, further research should explore the effects of the parasite serpin on this process.

Flavonoid-Quinoxaline Hybrid Compounds as Cathepsin Inhibitors Against Fascioliasis.

Fasciola hepatica is a parasitic trematode that infects livestock animals and humans, causing significant health and economic burdens worldwide. The extensive use of anthelmintic drugs has led to the emergence of resistant parasite strains, posing a threat to treatment success. The complex life cycle of the liver fluke, coupled with limited funding and research interest, have hindered progress in drug discovery. Our group has been working in drug development against this parasite using cathepsin proteases as molecular targets, finding promising compound candidates with in vitro and in vivo efficacy. Here, we evaluated hybrid molecules that combine two chemotypes, chalcones and quinoxaline 1,4-di- N-oxides, previously found to inhibit F. hepatica cathepsin Ls and tested their in vitro activity with the isolated targets and the parasites in culture. These molecules proved to be good cathepsin inhibitors and to kill the juvenile parasites at micromolar concentrations. Also, we performed molecular docking studies to analyze the compounds-cathepsins interface, finding that the best inhibitors interact at the active site cleft and contact the catalytic dyad and residues belonging to the substrate binding pockets. We conclude that the hybrid compounds constitute promising scaffolds for the further development of new fasciolicidal compounds.

A multidisciplinary analysis of over 53,000 fascioliasis patients along the 1995–2019 countrywide spread in Vietnam defines a new epidemiological baseline for One Health approaches

Fascioliasis, only foodborne trematodiasis of worldwide distribution, is caused by Fasciola hepatica and F. gigantica, liver flukes transmitted by freshwater snails. Southern and southeastern Asia is an emerging hot spot of F. gigantica, despite its hitherto less involvement in human infection. In Vietnam, increasing cases have been reported since 1995, whereas only sixteen throughout 1800–1994. A database was created to include epidemiological data of fascioliasis patients from the 63 Vietnam provinces throughout 1995–2019. Case profiles were based on serology, symptoms, eosinophilia, imaging techniques, stool egg finding, and post-specific-treatment recovery. Radio broadcasting about symptoms and costless diagnosis/treatment led patients to hospitals after symptom onset. Yearly case numbers were modelled and spatio-temporally analyzed. Missing data and confounders were assessed. The countrywide spread has no precedent. It started in the central coast, including 53,109 patients, mostly adults and females. Seasonality, linked to vegetable consumption, peaks in June, although the intensity of this peak differs according to relief/climatic zones. Incidence data and logistic regression curves are obtained for the first time in human fascioliasis. Fasciolid hybrids accompanying the spreading F. gigantica flukes, and climate change assessed by risk index correlations, are both ruled out as outbreak causes. Human-guided movements of livestock from an original area prove to be the way used by fasciolids and lymnaeid vectors to expand geographically. Radix viridis, a highly efficient transmitting and colonizing vector, played a decisive role in the spread. The use of irrigated crop fields, widely inhabited by R. viridis, for livestock grazing facilitated the transmission and spread of the disease. General physician awareness and diagnostic capacity improvement proved the successful impact of such knowledge transfer in facilitating and increasing patient infection detection. Information, education and communication to the public by radio broadcasting demonstrated to be very helpful. Fasciola gigantica is able to cause epidemic and endemic situations similar to F. hepatica. The magnitude of the human outbreak in Vietnam is a health wake-up call for southern and southeastern countries of Asia which present the highest human population densities with increasing food demands, uncontrolled livestock inter-country exchange, foreign import practices, and monsoon's increasing climate change impact.

Climatic variations and Fasciola: a review of impacts across the parasite life cycle.

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

Advancing vaccine development against Opisthorchis viverrini: A synergistic integration of omics technologies and advanced computational tools.

The liver fluke O. viverrini (Opisthorchis viverrini), a neglected tropical disease (NTD), endemic to the Great Mekong Subregion (GMS), mainly afflicts the northeastern region of Thailand. It is a leading cause of cholangiocarcinoma (CCA) in humans. Presently, the treatment modalities for opisthorchiasis incorporate the use of the antihelminthic drug praziquantel, the rapid occurrence of reinfection, and the looming threat of drug resistance highlight the urgent need for vaccine development. Recent advances in "omics" technologies have proven to be a powerful tool for such studies. Utilizing candidate proteins identified through proteomics and refined via immunoproteomics, reverse vaccinology (RV) offers promising prospects for designing vaccines targeting essential antibody responses to eliminate parasite. Machine learning-based computational tools can predict epitopes of candidate protein/antigens exhibiting high binding affinities for B cells, MHC classes I and II, indicating strong potential for triggering both humoral and cell-mediated immune responses. Subsequently, these vaccine designs can undergo population-specific testing and docking/dynamics studies to assess efficacy and synergistic immunogenicity. Hence, refining proteomics data through immunoinformatics and employing computational tools to generate antigen-specific targets for trials offers a targeted and efficient approach to vaccine development that applies to all domains of parasite infections. In this review, we delve into the strategic antigen selection process using omics modalities for the O. viverrini parasite and propose an innovative framework for vaccine design. We harness omics technologies to revolutionize vaccine development, promising accelerated discoveries and streamlined preclinical and clinical evaluations.

Construction and mouse antibody response evaluation of juvenile stage-specific chimeric protein from Fasciola gigantica

Fasciolosis, caused by the liver fluke Fasciola gigantica, is a major parasitic disease that affects livestock and therefore causes significant economic losses in tropical countries. Although anthelminthic drugs can kill the parasite, drug-resistant liver fluke populations are increasing. In this study, a recombinant F. gigantica chimeric protein (rFgCHI) consisting of cathepsin L1H (FgCL1H), cathepsin B3 (FgCB3), and Saposin-like protein 1 (FgSAP1) was designed and expressed in Escherichia coli (BL21). The molecular weight of rFgCHI was 61 kDa. To study the antibody response, male BALB/c mice were immunized via the subcutaneous injection of rFgCHI combined with Quil A. Immunization with rFgCHI showed the induction of IgG1 and IgG2a with a higher IgG1 isotype level, indicating the potential of mixed Th1/Th2 immune responses, with Th2 predominating. However, the results showed high levels of IgG against the single proteins, except for rFgSAP1. Through Western blotting, mouse anti-rFgCHI polyclonal antibodies could be detected to the native proteins obtained from the parasite at all stages. Immunolocalization also revealed that the anti-rFgCHI antibodies could detect targeted antigens in the cecal epithelium of the parasite. These results demonstrated that rFgCHI is immunogenic to the mouse immune system and may potentially be a protein candidate for the development of a fasciolosis vaccine.

Prevalence of Bovine Fasciolosis with Fasciola Hepatica in the M'pila Slaughter Areas of Congo Brazzaville

Fasciolosis is a hepatobiliary dystomatosis due to the migration into the liver parenchyma of immature form of parasites called fasciola hepatica or liver fluke. It is a cosmopolitan disease which represents economic and health problems whose diagnosis is rarely made, it is usually a discovery at the slaughterhouse. It mainly affects animals and sometimes humans. It is in this context that this study was carried out at two slaughterhouses in Mpila in order to determine the prevalence of bovine fasciolosis according to breeds, age, sex, origin and slaughter sector. The liver incision technique to search for flukes was carried out on 356 cattle. The results of the analysis carried out on 356 cattle including 134 infested cattle showed a prevalence of 37.64%. Furthermore, the highest prevalence was observed among mixed race people, a rate of 53.84%. Thus, cattle aged over 4 years were the most exposed to this pathology. Our results show that fasciolosis is a reality in the slaughter areas of Brazzaville. Monitoring of this pathology must be carried out to protect the health of our animals and that of humans.

Glucose transporters and sodium glucose co-transporters cooperatively import glucose into energy-demanding organs in carcinogenic liver fluke Clonorchis sinensis.

The liver fluke Clonorchis sinensis imports large amounts of glucose to generate energy and metabolic intermediates through glycolysis. We hypothesized that C. sinensis absorbs glucose through glucose transporters and identified four subtypes of glucose transporter (CsGTP) and one sodium glucose co-transporter (CsSGLT) in C. sinensis. Expressed sequence tags encoding CsGTPs were retrieved from the C. sinensis transcriptome database, and their full-length cDNA sequences were obtained by rapid amplification of cDNA ends (RACE). The tissue distribution of glucose transporters in C. sinensis adults was determined using immunohistochemical staining. Developmental expression was measured using RT-qPCR. The transport and distribution of glucose into living C. sinensis were monitored using confocal microscopy. Membrane topology and key functional residues of CsGTPs were homologous to their counterparts in animals and humans. CsGTP1, 2, and 4 were transcribed 2.4-5.5 times higher in the adults than metacercariae, while CsGTP3 was transcribed 2.1 times higher in the metacercariae than adults. CsSGLT transcription was 163.6 times higher in adults than in metacercariae. In adults, CsSGLT was most abundant in the tegument; CsGTP3 and CsSGLT were localized in the vitelline gland, uterine wall, eggs, mesenchymal tissue, and testes; CsGTP4 was found in sperm and mesenchymal tissue; and CsGTP1 was mainly in the sperm and testes. In C. sinensis adults, exogenous glucose is imported in a short time and is present mainly in the middle and posterior body, in which the somatic and reproductive organs are located. Of the exogenous glucose, 53.6% was imported through CsSGLT and 46.4% through CsGTPs. Exogenous glucose import was effectively inhibited by cytochalasin B and phlorizin. We propose that CsSGLT cooperates with CsGTPs to import exogenous glucose from the environmental bile, transport glucose across mesenchymal tissue cells, and finally supply energy-demanding organs in C. sinensis adults. Studies on glucose transporters may pave the way for the development of new anthelmintic drugs.

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.

Mitochondrial genome of the fluke pond snail, Austropeplea cf. brazieri (Gastropoda: Lymnaeidae)

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

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.

A Severe Clinical Case of Theileriosis with Terminal Wasting in Adult Cattle: A Veterinary Clinical and Pathology Case Report

This veterinary clinical and pathology case report describes a severe clinical case of theileriosis in adult cattle with prolonged recumbency and terminal wasting because of complications due to chronic theileriosis in two adult (2-year-old) Jersey-Friesian Cross. The case was referred to the large animal clinic of the University Veterinary Hospital (UVH), Faculty of Veterinary Medicine, Universiti Putra Malaysia (UPM). The animals were presented as recumbent, dull, depressed, hypothermic, dehydrated, and cachexic. There were poor responses of the patients to fluid therapy and non-steroidal anti-inflammatory treatments, and they both died within 24 hrs of clinical presentation. Haematology (complete blood count and serum biochemistry) results revealed haemoconcentration and faecal examinations were negative for liver fluke infestation and blood smear examination was positive for intraerythrocytic piroplasms of Theileria (0.01% parasitaemia). Post-mortem examination further revealed advanced cachexia, generalised serous atrophy of fat, hepatomegaly, hepatic congestion, pulmonary congestion, and pulmonary oedema. Histological findings revealed atrophy of the hepatic cord, hepatic congestion, splenic hemosiderosis, pulmonary oedema and congestion. The clinicopathological findings supported an aetiological diagnosis of a severe clinical case of Theileriosis with terminal wasting because of complications due to chronic Theileriosis infection.

Evaluation of Reproductive Histology Response of Adult Fasciola hepatica in Goats Vaccinated with Cathepsin L Phage-Exposed Mimotopes.

Fasciolosis, a globally re-emerging zoonotic disease, is mostly caused by the parasitic infection with Fasciola hepatica, often known as the liver fluke. This disease has a considerable impact on livestock productivity. This study aimed to evaluate the fluke burdens and faecal egg counts in goats that were administered phage clones of cathepsin L mimotopes and then infected with F. hepatica metacercariae. Additionally, the impact of vaccination on the histology of the reproductive system, specifically related to egg generation in adult parasites, was examined. A total of twenty-four goats, which were raised in sheds, were divided into four groups consisting of six animals each. These groups were randomly assigned. The goats were then subjected to two rounds of vaccination. Each vaccination involved the administration of 1 × 1013 phage particles containing specific mimotopes for cathepsin L2 (group 1: PPIRNGK), cathepsin L1 (group 2: DPWWLKQ), and cathepsin L1 (group 3: SGTFLFS). The immunisations were carried out on weeks 0 and 4, and the Quil A adjuvant was used in combination with the mimotopes. The control group was administered phosphate-buffered saline (PBS) (group 4). At week 6, all groups were orally infected with 200 metacercariae of F. hepatica. At week 22 following the initial immunisation, the subjects were euthanised, and adult F. hepatica specimens were retrieved from the bile ducts and liver tissue, and subsequently quantified. The specimens underwent whole-mount histology for the examination of the reproductive system, including the testis, ovary, vitellaria, Mehlis' gland, and uterus. The mean fluke burdens following the challenge were seen to decrease by 50.4%, 62.2%, and 75.3% (p < 0.05) in goats that received vaccinations containing cathepsin L2 PPIRNGK, cathepsin L1 DPWWLKQ, and cathepsin L1 SGTFLFS, respectively. Animals that received vaccination exhibited a significant reduction in the production of parasite eggs. The levels of IgG1 and IgG2 isotypes in vaccinated goats were significantly higher than in the control group, indicating that protection is associated with the induction of a mixed Th1/Th2 immune response. The administration of cathepsin L to goats exhibits a modest level of efficacy in inducing histological impairment in the reproductive organs of liver flukes, resulting in a reduction in egg output.

Occurrence, ecology and management of Fascioloides magna in Bavaria, southern Germany

In 2015, red deer infected with American liver fluke were discovered in the Veldenstein Forest area for the first time. This was the first detection of the fluke in a wild deer population in Bavaria. The aim of the study was to determine the American liver fluke prevalence rates in red deer, roe deer and wild boar in the Veldenstein Forest, as well as factors influencing these rates. Since 2018, the livers of 83% of the red deer culled in the study area have been examined, as well as those of the occasional roe deer and wild boar. The livers are classified by adspection and dissection into four levels of infection. The age of the animals was estimated based on dentition and tooth cementum annuli in the first molar (M1). The livers of 520 red deer, 226 roe deer and 75 wild boar were dissected. All wild boar livers tested negative. 3% of roe deer and 36% of red deer livers were positive. The red deer population was reduced to lower the prevalence, beginning in 2018. In the following years, it fell significantly. Medium and high infection levels were initially detected in 61% of adult red deer. Since 2018, the median number of flukes per infected liver has decreased significantly from 17.51 flukes/liver to 10.0 flukes/liver. On driven hunts, significantly more diseased deer were found than during hunting from raised hides. Furthermore, there are close correlations between the age of the red deer, infections, and the distance from the Pegnitz floodplains. The American liver fluke leads to the extinction of roe deer locally and to a massive infection and decimation of the red deer. Without a drastic reduction in the number of infected deer populations, there is a risk of large‐scale infection of native deer species across Bavaria.

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.

Immunomodulatory effect of hemozoin from Opisthorchis felineus and its participation in reducing allergic inflammation through activation of the inflammasome

Molecules of the excretory-secretory product arising as a result of co-evolution of a host and a parasite are able to inhibit the type 2 immune response, while activating the type 1 and the type 17 responses. The ability to alter the immune response can be used to inhibit inflammation in allergic diseases. In this regard, a search for helminth-associated molecules both with an immunomodulatory effect and immunogenicity and low toxicity is represented as a topical task. Hemozoin being a dark brown insoluble biocrystal is an excretory product of a number of hematophagous parasites (Schistosoma mansoni, Plasmodium falciparum and Opisthorchis felineus). This substance as a compound of parasitic origin with explicit immunomodulatory properties has prospects for deep research. Lots of relevant literature has been studied carefully before developing a concept about Opisthorchis felineus hemozoin and its unique properties. Thus, the current review presents an analysis of the accumulated data regarding the effect of Opisthorchis felineus trematode on the host immune system. Also the data on the immunomodulatory effect of hemozoin of various origins is analyzed. The current knowledge on the immune mechanisms of inflammasome activation and the possible role of hemozoin in reducing allergic inflammation through this mechanism is represented in this article. According to the systemized research results the excretory-secretory molecule of the liver fluke is a product of parasitic origin with an explicit immunomodulatory effect which is promising for prospective scientific study. Opisthorchis felineus extract increases the expression of T-regulatory cells and inhibits the Th2-immune response. Hemozoin produced by Opisthorchis felineus, like one produced by Plasmodium falciparum, may be involved in reducing the activity of allergic inflammation through activation of the inflammasome. Comprehension of the mechanism of how hemozoin interacts with the host immune system can be applied for correction of conditions associated with Th2-polarization of the immune response, which primarily include atopic diseases. Studying the mechanism of inflammation will help by the search for a biological target to create a vaccine to prevent the spread of atopic diseases, including bronchial asthma.