Exploring glutathione transferase and Cathepsin L-like proteinase for designing of epitopes-based vaccine against Fasciola hepatica by immunoinformatics and biophysics studies.

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

Investigation of Fasciola gigantica in freshwater snail Radix (Lymnaea) spp. In the highly parasite-prevalent area of Nakhon Ratchasima Province, Thailand.

This study investigates the distribution of the Lymnaea (Radix) spp. in Pak Chong district, Nakhon Ratchasima province, northeast Thailand, where a vast cattle farming area is located and has a high prevalence of Fasciola spp. in the cattle. By random selection, 1,414 snails were collected from the natural and man-made ponds. The snails were recorded for morphology and processed for DNA isolation. The snail species were investigated by conventional PCR using a 16S rDNA-specific primer. The result demonstrated that all collected snails were R. (L.) rubiginosa. Moreover, the infection of Fasciola gigantica in the snails was investigated by PCR using a cytochrome c oxidase I (COX1)-specific primer. The results illustrated that the overall prevalence was 22.5% (318/1414), with the highest prevalence in the Nong Sa Rai subdistrict at 73.6% (81/110), which is the highest prevalence of Fasciola gigantica in the snail host that has ever been reported. The lowest prevalence existed in the Pong Ta Long subdistrict at 3.7% (4/109). Our results corresponded to the previous report on the Fasciola spp. infection in the cattle from this area, and the geographical analysis revealed that the most suspected factor would be the earth dam located in these subdistricts, where many animals live freely during the day. Our findings could be helpful for further parasite control and could trigger the study of the biology and associated factors in the future.

The use of a Bayesian latent class model to estimate the test characteristics of three liver fluke diagnostic tests under New Zealand field conditions.

The liver fluke Fasciola hepatica is a trematode parasite of farmed livestock with worldwide distribution, causing chronic production losses and possible death from hepatobiliary damage. The effective management of liver fluke infection requires diagnostic tests which can accurately identify infected animals at both the individual and herd level. However, the accuracy of liver fluke diagnostic tests performed on individual New Zealand cattle is currently unknown. The aim of this study was to use a Bayesian latent class model (LCM) to estimate the test characteristics of three liver fluke diagnostic tests, the coproantigen ELISA, the IDEXX antibody ELISA and the faecal egg count. One hundred and twenty dairy cows each from two dairy farms were blood and faecal sampled in April 2021. The samples were transported to Massey University, Palmerston North, and the three diagnostic tests completed following the respective manufacturer instructions. A Bayesian LCM model, adapted from the original Hui and Walter 2 tests 2 populations model, was built to estimate the test characteristics of the three diagnostic tests in the two dairy herds. The model was implemented in JAGS using Markov chain Monte Carlo sampling. The first 30,000 iterations were discarded as burn-in, and the next 200,000 iterations were used to construct the posterior distributions. Uninformed priors, beta (1,1), were used as the prior distributions for the prevalence estimation and informed beta priors, based on published results, were used as the prior distributions for estimating the sensitivity and specificity of each diagnostic test. Model convergence was confirmed by inspection of trace plots and examination of the results of the Gelman and Rubin test. The results found that the coproantigen ELISA test was the most accurate for diagnosing liver fluke infection in individual animals with a sensitivity = 0.98 (95 % CI 0.95-1.00) and specificity = 0.95 (95 % CI 0.81-1.00) compared to the IDEXX antibody ELISA test, sensitivity = 0.39 (95 % CI 0.32-0.47) and specificity = 0.86 (95 % CI 0.75-0.96) or the FEC, sensitivity = 0.23 (95 % CI 0.17-0.30) and specificity = 0.92 (95 % CI 0.86-0.97). Based on these results clinicians should be encouraged to use the coproantigen ELISA test to diagnose liver fluke infection in individual cattle.

Helminth co-infections have no additive detrimental impact on milk yield and milk quality compared to mono-infections in German dairy cows.

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

Fosfatriclaben: Effective dose determination and comparative efficacy assessment with closantel, triclabendazole+ivermectin, triclabendazole+albendazole in artificially infected cattle.

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.

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.

Qualitative analysis of Fasciola gigantica excretory and secretory products coimmunoprecipitated with buffalo secondary infection sera shows dissimilar components from primary infection sera.

Buffaloes cannot mount a robust adaptive immune response to secondary infection by Fasciola gigantica. Even if excretory and secretory products (ESPs) exhibit potent immunoregulatory effects during primary infection, research on ESPs in secondary infection is lacking, even though the ESP components that are excreted/secreted during secondary infection are unknown. Therefore, qualitative analysis of ESP during secondary infection was performed and compared with that of primary infection to deepen the recognition of secondary infection and facilitate immunoregulatory molecules screening. Buffaloes were divided into three groups: A (n = 3, noninfected), B (n = 3, primary infection) and C (n = 3, secondary infection). Buffaloes in the primary (0 weeks post infection; wpi) and secondary (-4 and 0 wpi) infection groups were infected with 250 metacercariae by oral administration. Then, sera were collected from groups at different wpi, and interacting proteins were precipitated by coimmunoprecipitation (Co-IP), qualitatively analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and annotated by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to infer their potential functions. In group C, 324 proteins were identified, of which 76 proteins were consistently identified across 7 time points (1, 3, 6, 8, 10, 13, and 16 wpi). Compared with 87 proteins consistently identified in group B, 22 proteins were identified in group C. Meanwhile, 34 proteins were only identified in group C compared to 200 proteins identified in group B. Protein pathway analysis indicated that these proteins were mainly involved in the cellular processes and metabolism of F. gigantica. Among them, 14-3-3θ was consistently identified in group C and may be involved in various cellular processes and innate immune signalling pathways. Members of the HSP family were identified in both groups B and C and may function in both primary and secondary infection processes. The proteins discovered in the present study will help to deepen the understanding of the molecular interactions between F. gigantica and buffalo during secondary infection and facilitate the identification of new potential immunoregulatory molecules.

Both host and parasite non-coding RNAs co-ordinate the regulation of macrophage gene expression to reduce pro-inflammatory immune responses and promote tissue repair pathways during infection with fasciola hepatica.

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

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.

Molecular Characterization and Phylogenetic Analysis of Fasciola gigantica Based on ITS-1 Genetic Marker and copro-epidemiological Survey from Ruminants of Pakistan.

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

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.

Unanticipated discovery: incidental encounter with fasciola hepatica during ERCP - a case report.

Introduction and Importance: Fasciola hepatica (FH) is a rare parasitic infection in humans. Its incidental detection during endoscopic retrograde cholangiopancreatography (ERCP) is exceptionally uncommon. This case underscores the importance of considering parasitic infections, even in low-endemicity regions, and the potential implications of dietary and environmental factors in disease transmission. Case Presentation: The authors present a case of a 31-year-old female from Dhading, Nepal, who underwent ERCP for suspected biliary stone. The patient had been experiencing recurring, nonradiating, burning epigastric pain for 5 to 7 years, which had recently intensified. Previous evaluations, including abdominal ultrasonography, CT, and MRI, revealed a dilation within the common bile duct and an obstruction in the biliary system. Clinical Discussion: During ERCP, cholangiography revealed mildly dilated extra and intrahepatic bile ducts with irregular filling defects in the common hepatic duct. Sphincterotomy was performed, followed by the extraction of multiple FH worms. A 7 Fr 7 cm double pigtail plastic stent was placed with a good flow of bile. However, the patient experienced anaphylaxis during the procedure, necessitating swift and tailored administration of appropriate medications to ensure effective management and stabilization. The patient was closely monitored in the ICU postprocedure. Conclusion: After careful monitoring and treatment, the patient fully recovered. The unexpected discovery of FH during ERCP is extremely rare. Early recognition and appropriate management of such incidental findings are crucial to ensuring optimal patient outcomes.

Anatomical and electrophysiological analysis of the fasciola cinerea of the mouse hippocampus.

The hippocampus is considered essential for several forms of declarative memory, including spatial and social memory. Despite the extensive research of the classic subfields of the hippocampus, the fasciola cinerea (FC)-a medially located structure within the hippocampal formation-has remained largely unexplored. In the present study, we performed a morpho-functional characterization of principal neurons in the mouse FC. Using in vivo juxtacellular recording of single neurons, we found that FC neurons are distinct from neighboring CA1 pyramidal cells, both morphologically and electrophysiologically. Specifically, FC neurons displayed non-pyramidal morphology and granule cell-like apical dendrites. Compared to neighboring CA1 pyramidal neurons, FC neurons exhibited more regular in vivo firing patterns and a lower tendency to fire spikes at short interspike intervals. Furthermore, tracing experiments revealed that the FC receives inputs from the lateral but not the medial entorhinal cortex and CA3, and it provides a major intra-hippocampal projection to the septal CA2 and sparser inputs to the distal CA1. Overall, our results indicate that the FC is a morphologically and electrophysiologically distinct subfield of the hippocampal formation; given the established role of CA2 in social memory and seizure initiation, the unique efferent intra-hippocampal connectivity of the FC points to possible roles in social cognition and temporal lobe epilepsy.

Characteristics of Hepatobiliary Fascioliasis and the Role of Endoscopic Retrograde Cholangiopancreatography in Management: A Single Center Experience.

Background: Hepatobiliary fascioliasis has two phases, each requiring specific management approaches. Triclabendazole has been widely effective in treating the two phases of clinical fascioliasis and endoscopic retrograde cholangiopancreatography (ERCP) in the biliary phase. We aimed to characterize presentations of hepatobiliary fascioliasis and highlight the role of ERCP in management. Subjects and methods: This retrospective cohort includes patients diagnosed with clinical hepatobiliary fascioliasis between January 2013 and December 2022. Demographic data, clinical presentation, laboratory and radiological investigations, treatment, and endoscopy reports were collected from the records of 62 participants. Patients were divided into two groups: acute hepatic and chronic biliary phases. Results: Thirty-six patients were in the biliary phase, and 26 were in the hepatic phase. All patients were from rural areas, and females were predominant (76%). Hypereosinophilia was detected in 92% of acute cases and 58% of chronic biliary cases. In chronic biliary cases, the levels of liver biochemicals, including alanine transaminase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), and bilirubin, were higher at levels of 189 ± 76, 127 ± 47, 268 ± 77, and 2.4 ± 0.7 respectively, compared to acute hepatic cases, 35.6 ± 8.2, 32.7 ± 4.3, 69.2 ± 45.45, and 0.58 ± 0.01. The corresponding P-values were 0.003, 0.001, <0.001, and <0.001, respectively. Triclabendazole effectively cured 93.5% of patients and was used in combination with ERCP in biliary-phase cases where the fluke was extracted from the biliary system in 34 patients (94.4%). Three patients (8.8%) were diagnosed with post-ERCP pancreatitis. None of the patients experienced bleeding, perforation, or required biliary stenting. Conclusion: Clinical fascioliasis could manifest in acute hepatic or chronic biliary phases. Hypereosinophilia was more evident in the hepatic phases, while ALT, AST, GGT, and bilirubin were higher in the biliary phase. Triclabendazole is effective in the hepatic phase and when combined with ERCP in the biliary phase. ERCP is highly effective for relieving obstruction and treating biliary fascioliasis.

Fasciola hepatica: Updates on egg morphology, host range, and distribution.

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.

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.

The Hepatic Phase of Fascioliasis Complicated with Liver Bleeding: Presentation of Three Cases.

Bleeding is a very rare complication of Fasciola species (F. hepatica or F. gigantica) infection. We present here three cases of subcapsular liver bleeding caused by the hepatic phase of Fasciola spp. infection in patients, two of whom were women, aged 22, 66, and 84 years in Diyarbakir Province, southeastern Turkey. They had symptoms of right upper quadrant pain (n = 3), nausea (n = 1), and vomiting (n = 2) for periods ranging from 6 hours to 15 days. All patients with clinical presentations ranging from moderate abdominal pain to hypovolemic shock and ischemic hepatitis were improved with supportive treatment without the need for surgery. They showed complete clinical and laboratory recovery after triclabendazole administration in their follow-up. In conclusion, Fasciola spp. infection should be considered in the etiology of bleeding from liver disease.

Diagnosis and Treatment of Fasciola Infection with Endoscopic Retrograde Cholangiopancreatography (ERCP) in Ardabil, Northwest of Iran: Two Case Reports.

Here we report biliary fascioliasis in two women 27 and 54 years old from Ardabil Province, Iran who presented with nausea, anorexia, weight loss, and pain between shoulder blades. Endoscopic ultrasound showed distal common bile duct strictures (CBD) and a lesion in its proximal. Endoscopic retrograde cholangiopancreatography (ERCP) was performed and live parasites were diagnosed and successfully managed into the duodenal lumen. The clinical findings of the patient improved after the procedure. This report emphasizes that the prevalence of Fasciola in Ardabil is predictable and this infection could be simultaneously diagnosed and treated by the ERCP.