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

Fasciolosis is a food and waterborne disease caused by Fasciola spp., representing a global health burden to various hosts, including humans and other animals. This study investigates the in vitro activity of tellurium- and selenium-containing diaryl dichalcogenides: diacetal ditelluride (LQ07), diacetal diselenide (LQ62), and diacetyl diselenide (LQ68) alone and in combination with ivermectin (IVM) against eggs of Fasciola hepatica. The eggs were exposed for 12 h with each organochalcogen (OC) (0.1 - 2 mmol l -1 ) and IVM (0.01 - 2 mmol l -1 ) following an incubation of 15 days, allowing embryonation. The inhibitory concentration of 50 % (IC 50 ) of each OC or IVM was tested with the IC 10 , IC 30 , and IC 50 of IVM or each OC, respectively. LQ07, LQ62, and LQ68, as well as IVM, demonstrated a concentration-dependent ovicidal activity. The peak ovicidal activity of 99.74 % was achieved when IVM was tested at 2.0 mmol l -1 . LQ62 and LQ68 demonstrated greater ovicidal activity, having an IC 50 < 0.32 mmol l -1 being 6.25-fold more toxic than IVM alone. The percentage of dead eggs was significantly higher in the IVM group (early mortality), as Se-containing OCs led to the (miracidia) embryonation of the eggs with no hatching (late mortality). Blending Se-containing OCs and IVM showed an additive effect of up to 27 % against F. hepatica eggs. The present data contribute to the potential use of blending-based therapeutic strategies to combat F. hepatica infections in eradication programs worldwide. The combinations may also act against multidrug-resistant strains, reinstating drug-based parasite control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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., (© 2024 Wiley-VCH GmbH.)

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., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Fasciolosis caused by Fasciola hepatica is a major public health and economic problem worldwide. Due to the lack of a successful vaccine and emerging resistance to the drug triclabendazole, alternative phytotherapeutic approaches are being investigated. This study investigated the in vitro anthelmintic activity of Lavender (Lavandula angustifolia) and carob (Ceratonia siliqua L.) essential oils (EOs) against F. hepatica. The in vitro study was based on an egg hatch assay (EHA), adult motility inhibition assays, DNA damage, reactive oxygen species (ROS) level along with several oxidative stress biomarkers including glutathione peroxidase (GSH), and glutathione-S-transferase (GST), superoxide dismutase (SOD) and malondialdehyde (MDA). To this end, different concentrations of L. angustifolia and C. siliqua EOs (1, 5, 10, 25 and 50 mg/mL) were used to assess anthelmintic effects on different life stages including egg, and adults of F. hepatica for 24 hrs. The results indicated that these EOs play a significant role as anthelminthics, and the effect was dependent on time and concentration. The in vitro treatment of F. hepatica worms with both L. angustifolia and C. siliqua EOs increased DNA damage, ROS production and induction of oxidative stress (decreased SOD, GST and GSH, and increased MDA), significantly compared to control. Therefore, it can be concluded that L. angustifolia and C. siliqua EOs have the potential to be used as novel agents for the control and treatment of F. hepatica infections. Further studies are required to investigate their pharmacological potential and effectiveness in vivo for the treatment of parasitic infections., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Allahyari et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Fascioliasis is a neglected tropical zoonotic disease caused by liver flukes belonging to the genus Fasciola . The emergence of resistance to triclabendazole, the only World Health Organization-recommended drug for this disease, highlights the need for the development of new drugs. Helminths possess an anaerobic mitochondrial respiratory chain (fumarate respiration) which is considered a potential drug target. This study aimed to evaluate the occurrence of fumarate respiration in Fasciola flukes. We analyzed the properties of the respiratory chain of Fasciola flukes in both adults and newly excysted juveniles (NEJs). Fasciola flukes travel and mature through the stomach, bowel, and abdominal cavity to the liver, where oxygen levels gradually decline. High fumarate reductase activity was observed in the mitochondrial fraction of adult Fasciola flukes. Furthermore, rhodoquinone-10 (RQ 10 Em'= -63 mV), a low-potential electron mediator used in fumarate respiration was found to be predominant in adults. In contrast, the activity of oxygen respiration was low in adults. Rotenone, atpenin A5, and ascochlorin, typical inhibitors of mitochondrial enzymes in complexes I, II, and III, respectively, inhibit the activity of each enzyme in the adult mitochondrial fraction. These inhibitors were then used for in vitro viability tests of NEJs. Under aerobic conditions, NEJs were killed by rotenone or ascochlorin, which inhibit aerobic respiration (complex I-III), whereas atpenin A5, which inhibits complex II involved in fumarate respiration, did not affect NEJs. Moreover, ubiquinone-10 (UQ 10 Em'= +110 mV), which is used in oxidative respiration, was detected in NEJs, in addition to RQ 10 . In contrast, under anaerobic conditions, rotenone and atpenin A5, which inhibit fumarate respiration (complex I-II), were crucial for NEJs. These findings demonstrate that NEJs have active hybrid respiration, in which they can properly use both oxygen and fumarate respiration, depending on oxygen availability. Thus, fumarate respiration is a promising drug target for Fasciola flukes, because it plays an essential role in both adults and NEJs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Tashibu, Inaoka, Sakamoto, Murakami, Zannatul, Kita and Ichikawa-Seki.)

Fasciolosis is an important zoonotic disease caused by the trematode Fasciola hepatica, and it causes great losses in bovine production. The anthelmintic resistance is a major problem in the control of fasciolosis. In this study, the F. hepatica egg development and hatching test (EDHT) was used for the evaluation of the ovicidal activity of commercial drugs, commonly used for treating infected cattle, which reflects F. hepatica anthelminthic resistance in infected bovines, according to recent literature. Bile samples from F. hepatica naturally parasitized cattle were obtained from slaughterhouses in the cities of Lages and Otacílio Costa, Santa Catarina State, Brazil. The bile was washed, the eggs were recovered, quantified, and distributed in universal collectors, with a minimum of 1,000 eggs per vial. Four commercial drugs were used in this study, containing albendazole sulfoxide (ABDZ), closantel (CSTL), nitroxynil (NTXL), and triclabendazole with fenbendazole (TBZF). The drugs were diluted according to the manufacturer instructions. All drugs, and the respective control, were tested in triplicates, with the quantity of recovered eggs determining the number of drugs to be tested. The vials were incubated for 28 days at 27 °C, and the eggs were classified according to their degree of development under a stereomicroscope. In total, 121 egg samples were analyzed. Two samples were identified as resistant to TBZF. Undetermined resistance/susceptibility has been found in two isolates treated with ABDZ, one treated with NTXL and six treated with TBZF. CSTL did not present ovicidal activity and cannot be used in EDHT. This is the first time that commercial drugs were used in F. hepatica EDHT., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

About a third of the world population is infected by helminth parasites implicated in foodborne trematodiasis. Fascioliasis is a worldwide disease caused by trematodes of the genus Fasciola spp. It generates huge economic losses to the agri-food industry and is currently considered an emerging zoonosis by the World Health Organization (WHO). The only available treatment relies on anthelmintic drugs, being triclabendazole (TCBZ) the drug of choice to control human infections. The emergence of TCBZ resistance in several countries and the lack of an effective vaccine to prevent infection highlights the need to develop new drugs to control this parasitosis. We have previously identified a group of benzochalcones as inhibitors of cathepsins, which have fasciolicidal activity in vitro and are potential new drugs for the control of fascioliasis. We selected the four most active compounds of this group to perform further preclinical studies. The compound's stability was determined against a liver microsomal enzyme fraction, obtaining half-lives of 34-169 min and low intrinsic clearance values (<13 μL/min/mg), as desirable for potential new drugs. None of the compounds were mutagenic or genotoxic and no in vitro cytotoxic effects were seen. Compounds C31 and C34 showed the highest selectivity index against liver fluke cathepsins when compared to human cathepsin L. They were selected for in vivo efficacy studies observing a protective effect, similar to TCBZ, in a mouse model of infection. Our findings strongly encourage us to continue the drug development pipeline for these molecules., Competing Interests: Declaration of competing interest None, (Copyright © 2023. Published by Elsevier Inc.)

Fasciola gigantica is one of the worldwide parasites that cause livestock and human illnesses. Chemotherapy is now the primary therapeutic option for its treatment. Drug abuse has led to the emergence of drug-resistant strains. As a result, there is an urgent need to discover natural and efficient anthelmintics against Fasciola spp. The study aims to evaluate the ovicidal activities of camel milk and its fractions on F. gigantica eggs. In the in vitro assay of F. gigantica eggs were submitted to different concentrations (0.5% and 1%) of camel milk fractions; Camel Milk Whey (CMW), Camel Milk Casein (CMC), and Skimmed Camel Milk (SCM) as well as a positive control (PC) of Nitroxynil (100 mg/ml) and a negative control (NC) with physiological saline. The Egg Hatching Assay (EHA) results showed that camel milk fractions exhibited ovicidal activity, especially CMW, and CMC, which showed 97.58 ± 0.58 and 96.9 ± 1.99 ovicidal activity, respectively, at a concentration of 1% after 15 days of treatment compared to PC, which exhibited 91.75 ± 4.95 ovicidal activity. The egg hatching ratios were 1.67% and 2.33% for CMW and CMC, respectively, compared to 70.17% for the NC and 6% for the PC. The LC 50 values for CMW and CMC on the 15th day of treatment were 0.20 and 9.13, respectively. From the results above, we can infer that camel milk and its fractions are promising as a new alternative for fascioliasis control., (© 2023. The Author(s).)

Fasciola hepatica infections lead to severe health problems and production losses in sheep farming, if not treated effectively. Triclabendazole has been used extensively over decades due to its unique efficacy range against all definitive hostfluke stages but published data about the susceptibility of F. hepatica to anthelmintics in Germany are lacking. This study aimed to identify current F. hepatica infections in German sheep flocks by coproscopic examinations and to evaluate the efficacy of anthelmintics with a focus on triclabendazole in a field study conducted from 2020 to 2022. Initial screening included 71 sheep farms, many of them with known history of fasciolosis. In this highly biased sample set, the frequency of F. hepatica infection at individual sheep and farm level were 12.8% and 35.2%, respectively. Additionally, eggs of Paramphistominae were found at frequencies of 4.8% and 15.5% at individual sheep and farm level, respectively. Due to low egg shedding intensity, faecal egg count reduction (FECR) tests could only be conducted on a few farms. The efficacy of triclabendazole was tested on 11 farms and albendazole on one farm, including 3-53 sheep/farm. Individual faecal samples were collected before and two weeks after treatment to evaluate the FECR using the sedimentation or FLUKEFINDER® or a modified FLUKEFINDER® method. On all farms a coproantigen reduction test was conducted in parallel. Lacking efficacy of triclabendazole even at double dosage was shown on one farm associated with a high number of animal losses due to acute fasciolosis. On this farm, the Fasciola miracidium development test was additionally performed, revealing a high in vitro ovicidal activity of albendazole while closantel was effective in vivo. On all other farms, sufficient efficacy of triclabendazole was observed. In conclusion, triclabendazole resistance appears not to be widespread on German sheep farms but, when present, can have serious effects on animal health., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

BACKGROUND Human fascioliasis is an emerging zoonotic disease caused by the trematodes, or flatworms, Fasciola hepatica and Fasciola gigantica, also known as liver flukes. This retrospective study aimed to report the epidemiological findings in 95 cases of human fascioliasis in Dali, Yunnan Province, southwestern China, diagnosed between 2012 and 2021. MATERIAL AND METHODS The epidemiologic and clinical data of 95 patients diagnosed with human fascioliasis in Dali area from January 2012 to December 2021 were collected and retrospectively analyzed. The diagnosis of fascioliasis was based on the Chinese National Standard of Diagnosis of Fascioliasis (WS/T566-2017). RESULTS The mean age of patients was 38.54±15.68 years, and there were more female patients than male (61.05% vs 38.95%). The high-incidence seasons were identified as summer and autumn. The patients with human fascioliasis lived in pastoral areas or were infected F. gigantica by consuming contaminated vegetables or water containing metacercaria. Meanwhile, human fascioliasis was diagnosed by positive serologic tests (1: 640), and Fasciola eggs (144-180×73-96 μm) were detected in stool samples of 6 patients. The most common clinical features were abdominal pain (70.53%), accompanied by elevated eosinophils in 89.5% of these patients. Antiparasitic treatment with triclabendazole at 10 mg/kg/day for 2 days led to symptom relief in all patients. CONCLUSIONS The findings from this observational epidemiological study have highlighted the importance of recognizing, diagnosing, and managing fascioliasis, which is an emerging zoonosis associated with increased human proximity to plant-eating domestic and farmed animals.

Hepatobiliary fascioliasis is one of the rare but important parasitic infection in endemic areas such as Pakistan. Clinical manifestations overlap with common clinical conditions such as hepatitis and biliary stones causing common bile duct obstruction. Fascioliasis is a zoonotic infection caused by a leaf-shaped organism known as Fasciola Hepatica. Humans are accidentally infected by consuming contaminated water or food, especially undercooked cattle and sheep liver or raw green vegetables. We report three cases with similar clinical manifestations presenting in two different cities in Pakistan. After detailed clinical evaluation and laboratory investigations, the diagnosis was established by abdominal ultrasound and diagnostic and therapeutic cholangiography. Patients were followed till complete resolution of their symptoms and normalisation of liver function tests.

In the fall of 2022, decreased triclabendazole (TCBZ) efficacy against F. hepatica was suspected in a sheep farm located in the Santa Cruz province, Argentinian Patagonia. Since TCBZ-resistance in F. hepatica has never been reported in this province, this study aimed to confirm potential TCBZ-resistance in F. hepatica and to evaluate the efficacy of closantel (CLO) and nitroxinil (NTX), through faecal egg count reduction test (FECRT), and the efficacy of albendazole (ABZ) through the in vitro egg hatch test (EHT) in sheep. Sixty-eight (68) animals were selected from a herd of eighty (80) female Merino naturally infected with F. hepatica based on eggs per gram of F. hepatica (EPGFh) counts and assigned into four (4) groups (n = 17 per group): Group Control, animals did not receive anthelmintic treatment; Group TCBZ, animals were orally treated with TCBZ (12 mg/kg); Group CLO, animals were orally treated with CLO (10 mg/kg); and Group NTX, animals were subcutaneously treated with NTX (10 mg/kg). The fluke egg output was monitored on days 0 and 21 post-treatment. For the EHT, liver fluke eggs were isolated from faecal samples (approx. 50 g) collected from animals of the control group. TCBZ efficacy against liver fluke was 53.4%, confirming the presence of TCBZ-resistant isolates on the farm. CLO and NTX were highly effective (100%) for the treatment of F. hepatica on this farm. The EHT was carried out in two different laboratories, in which was observed an ABZ efficacy of 95.8 (Bariloche) and 96.5% (Tandil). These results indicate the ABZ susceptibility of this F. hepatica isolate and the inter-laboratory precision of the test., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

There is a lack of epidemiological data on fascioliasis in Egypt regarding disease characteristics and treatment outcomes across different governorates. We aimed to identify the demographic, epidemiologic, clinical, laboratory, and radiological characteristics and treatment outcomes of patients diagnosed with fascioliasis in Egypt. Data on human fascioliasis were collected retrospectively from patients' medical records in the period between January 2018 and January 2020. The study included 261 patients. More than 40% of enrolled patients were in the age group of 21-40 years old. Geographically, 247 (94.6%) were from Assiut Governorate with 69.3% were from rural areas. The most frequent symptoms were right upper quadrant pain (96.9%), and fever (80.1%). Eosinophilia was found in 250 cases (95.8%). Hepatic focal lesions were detected in 131 (50.2%); out of them 64/131 (48.9%) had a single lesion. All patients received a single dose of 10 mg/kg of triclabendazole, 79.7% responded well to a single dose, while in 20.3% a second ± a third dose of treatment was requested. After therapy, there was a reduction in leucocytes, Fasciola antibodies titer, eosinophilic count, bilirubin, and liver enzymes with an increase in hemoglobin level. According to our findings, a high index of suspicion should be raised in cases with fever, right upper abdominal pain, and peripheral eosinophilia, and further imaging workup is mandated to detect hepatic focal lesions. Prompt treatment by triclabendazole can serve as a standard-of-care regimen even for suspected cases., (© 2023. The Author(s).)

Fasciola hepatica, the liver trematode, infects ruminants and causes economic loss. Because parasites are developing resistance to commercial drugs, the negative effects of parasitism are increasing. In this study, we aimed to evaluate the efficacy of cumin (Cuminum cyminum) essential oil against F. hepatica eggs and adults. The eggs were incubated with eight concentrations of the essential oil (0.031125-4.15 mg/mL), and viable eggs were counted after 14 days and classified as embryonated or non-embryonated. Adult flukes were incubated in Roswell Park Memorial Institute medium to ensure their viability and then incubated in essential oil. They were observed for 24 h after treatment. The adults were assessed with the two lowest effective oil concentrations used in the ovicidal test. Three controls were used for both tests: nitroxynil, a negative control, and Tween®80. After incubation in oil, the adult specimens were processed for histological analysis and stained with hematoxylin-eosin. In addition, the oil was tested for cytotoxicity using Madin-Darby bovine kidney cells to assess any possible effect on them. The oil was effective in ovicidal and adulticidal inhibition of the trematode, with statistically significant results. All concentrations assessed in the ovicidal test were 100% effective. The adult test was effective within 15 h and inactivated all the specimens at the highest concentration evaluated (0.06225 mg/mL). Histological analysis showed that cumin essential oil resulted in marked areas of vacuolization. The spines showed no structural changes but were surrounded by microvesicles. These findings indicated that cumin oil could be a potential compound in the control of fasciolosis., Competing Interests: Declaration of competing interest The authors declare there are no conflicts of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Knowsley Safari (KS), Prescot, United Kingdom houses a variety of captive exotic ungulates. As part of their animal welfare plan, a prospective coprological survey was undertaken for liver fluke. In June 2021, 330 fecal samples, representative of 18 exotic ungulate species, were processed by sedimentation and filtration, with examination by coproscopy. Finding fascioliasis in all five vicuña alone, with fecal egg counts ranging from one to eight eggs per gram, anthelminthic treatment was attempted twice, with three coprological reviews. While the first anthelminthic treatment (oxyclozanide) was equivocal, the second anthelminthic treatment (triclabendazole) was proven effective upon two later follow-ups. An initial malacological survey of 16 freshwater sites in KS, first found Galba truncatula at two sites in June 2021, then upon more extensive searching subsequently within the vicuña's enclosure. It appears that F. hepatica was locally acquired, being the first report of fascioliasis within captive vicuñas in the United Kingdom. To develop a better fluke-management plan, regular coprological and malacological surveillance is justified, perhaps with molecular xenomonitoring of snails, alongside prompt administration of appropriate flukicide as required.

The intensive use of anthelmintic drugs to control Fasciola hepatica infections in dairy cattle has resulted in the emergence of anthelmintic resistance. Cases of resistance to triclabendazole (TCBZ) have been reported worldwide. The main goal of this research was to evaluate the main five fasciolicides to control fasciolosis in dairy cattle in the Mantaro Valley, Peru. Two fecal egg count reduction tests were performed. In a first study, 24 naturally F. hepatica infected cattle were randomly grouped into three experimental groups (n = 8). Groups were treated with either TCBZ, nitroxynil (NTX) or closantel (CLOS). In a second experiment, 55 naturally infected cows were grouped into three experimental groups and treated with either TCBZ (n = 18), rafoxanide (RFX) + albendazole (ABZ) (n = 19) or clorsulon (CLN) + ivermectin (IVM) (n = 18). Therapeutic efficacy was determined following the WAAVP guidelines by measuring reduction in fluke egg output at days 15 and 30 post-treatment. Bootstrapping method was used to obtain the 95% confidence intervals. The efficacy of TCBZ was inadequate in both studies (≤80.8%). Closantel showed high efficacy (≥ 90%) at both days, while NTX showed 92.9% (83-100) and 82.1% (53.6-100), efficacy, at days 15 and 30, respectively. Efficacy for RFX were 92.1% (79.6-98.9) and 97.4% (94.1-99.4); and for CLN, 98.8% (97.6-100) and 80.1% (44.7-99.4), at days 15 and 30, respectively. The outcome of this study indicates reduced therapeutic efficacy of TCBZ against F. hepatica in an important dairy area of the Peruvian central highlands but also demonstrates the validity of four alternatives., Competing Interests: Declaration of Competing Interest Daniel Zarate Rendon reports article publishing charges, equipment, drugs, or supplies, and travel were provided by Flemish Interuniversity Council and the Universidad Nacional Agraria La Molina, Perú., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Helminth infections of cattle place significant burdens on livestock production and farm economic efficiency. Heavy infections are relatively easy to detect and treat with anthelmintics. However, subclinical infections have major but often hidden impacts on animals, necessitating more refined diagnostics to detect them and ideally inform farmers about the likely impact of anthelmintic treatment on animal and herd performance. Here, we review recent advances in diagnosing three major cattle helminth infections - gastrointestinal nematodes (GINs), liver flukes, and lungworms - and the search for subclinical infection thresholds to guide treatment decisions. Combining refined diagnostic thresholds with farm-specific information on grazing systems and animal history enables farmers to tailor helminth treatments to specific epidemiological circumstances, thereby limiting anthelmintic resistance (AR) and boosting agricultural efficiency and food security., Competing Interests: Declaration of interests J.C. and E.C. are involved in a license agreement between Ghent University and INDICAL Bioscience GmbH for commercialization of SVANOVIR® O. ostertagi-Ab and F. hepatica-Ab ELISA. The authors declare no other known conflicts of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Fasciola hepatica infection is responsible for substantial economic losses in livestock worldwide and poses a threat to human health in endemic areas. The mainstay of control in livestock and the only drug licenced for use in humans is triclabendazole (TCBZ). TCBZ resistance has been reported on every continent and threatens effective control of fasciolosis in many parts of the world. To date, understanding the genetic mechanisms underlying TCBZ resistance has been limited to studies of candidate genes, based on assumptions of their role in drug action. Taking an alternative approach, we combined a genetic cross with whole-genome sequencing to localise a ~3.2Mbp locus within the 1.2Gbp F. hepatica genome that confers TCBZ resistance. We validated this locus independently using bulk segregant analysis of F. hepatica populations and showed that it is the target of drug selection in the field. We genotyped individual parasites and tracked segregation and reassortment of SNPs to show that TCBZ resistance exhibits Mendelian inheritance and is conferred by a dominant allele. We defined gene content within this locus to pinpoint genes involved in membrane transport, (e.g. ATP-binding cassette family B, ABCB1), transmembrane signalling and signal transduction (e.g. GTP-Ras-adenylyl cyclase and EGF-like protein), DNA/RNA binding and transcriptional regulation (e.g. SANT/Myb-like DNA-binding domain protein) and drug storage and sequestration (e.g. fatty acid binding protein, FABP) as prime candidates for conferring TCBZ resistance. This study constitutes the first experimental cross and genome-wide approach for any heritable trait in F. hepatica and is key to understanding the evolution of drug resistance in Fasciola spp. to inform deployment of efficacious anthelmintic treatments in the field., Competing Interests: The authors declare that they have no competing interests., (Copyright: © 2023 Beesley et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A 38-year-old rice farmer visited a hospital for abdominal pain. Computed tomography (CT) showed a liver tumor and magnetic resonance imaging (MRI) revealed a hypovascular tumor, both in segment 4. Thus, he was diagnosed with liver abscess. Ten days later, CT showed a new liver tumor in segment 8, but the size of the liver tumor in segment 4 had decreased. He was suspected with parasitic disease because of eosinophilia. Enzyme-linked immunosorbent assay showed a high level of serum Fasciola antibody. The patient was diagnosed with fascioliasis and was treated with triclabendazole. Post-treatment, CT revealed that the liver tumors had shrunk. Eosinophilia and multiple lesions were characteristic findings of parasitic disease.

This is a case report of fascioliasis that progressed from the hepatic to the biliary phases over 2 years. A woman in her late 60s ate Zingiber mioga from the field, which was followed by abdominal pain that occurred 1 month later. Although CT and MRI studies revealed an increase in blood eosinophils as well as multiple hepatic nodules, they vanished quickly. After 2 years, an MRCP study revealed multiple flat lesions, which were diagnosed as adult fascioliasis. Definitive diagnosis was provided by enzyme-labeled antibody method using fasciola-specific antigen. Triclabendazole was administered once to complete the treatment.

Calicophoron daubneyi (rumen fluke) is an emerging parasitic infection of livestock across Europe. Despite increasing in prevalence, little is known about the level of awareness of rumen fluke or current control practices used by UK farmers. Fasciola hepatica (liver fluke) is a common parasitic infection of cattle and sheep in the UK. Co-infections with these parasites can present in sheep and cattle, but the only drug with reported efficacy against rumen fluke is oxyclozanide. Between December 2019 and March 2020, 451 sheep and/or cattle farmers completed an online questionnaire, capturing their awareness and current means of control for liver fluke and rumen fluke. Most respondents (70%) were aware of rumen fluke, with 14% recording its presence on their farms and 18% having previously treated for rumen fluke. Almost all respondents (99%) were aware of liver fluke and higher numbers of respondents reported its presence on farm (67%) with 88% having previously treated for liver fluke. Respondents who were aware of rumen fluke said they were concerned about the parasite (81%), although rumen fluke was less of a concern than liver fluke (p < 0.05). Of respondents who reported rumen fluke presence on their farm, 42% cited incorrect diagnostic methods, including those traditionally used to detect liver fluke. Respondents were more likely to treat annually for liver fluke, as opposed to rumen fluke (p < 0.05). The most frequently used drug for the treatment of liver fluke infection was triclabendazole (53% sheep treatments, 34% cattle treatments) and only a minority of farmers treated with a product effective against rumen fluke (oxyclozanide; 42% cattle treatments, 35% sheep treatments). A small proportion of farmers stated that they used a non-flukicide drug to treat sheep for liver fluke infection (1.6% sheep treatments). These results demonstrate a broad awareness of liver and rumen fluke in sheep and cattle, but reveal confusion amongst farmers about their diagnosis and treatment, highlighting the need to provide best practice advice to the livestock industry for the control of both parasites., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Fasciola gigantica , the causative agent of tropical fasciolosis, is a food-borne zoonotic trematode that affects around 80% livestock of Bangladesh. Triclabendazole (TCBZ), nitroxynil (NTON) and oxyclozanide (OCZN) are frequently used against fascioliasis; however, the current status of potency of these flukicides was unknown. In this study, in vitro efficacy of TCBZ, NTON and OCZN at various concentrations on F. gigantica has been evaluated by relative motility (RM), morphological distortions of apical cone through an inverted microscope, architectural and ultra-structural changes through histopathological and scanning electron microscopy (SEM). It is observed that TCBZ, NTON and OCZN at higher concentrations significantly ( P < 0.05) reduced RM of the flukes compared to untreated control. NTON at 150 μ g mL −1 was the most potent to reduce the motility within 4 h whereas TCBZ and OCZN were much delayed. Histopathological changes showed swollen, extensive cracking, numerous vacuoles and splitting of the tegument surrounding the spines; spine dislodged from its socket in treated flukes compared to untreated worms. Histopathological changes were more conspicuous at higher doses of TCBZ, NTON and OCZN. SEM has shown the disruption of the apical cone, apart from swelling of the tegument on the ventral surface corrugation and disruption of the ventral apical cone. All these changes indicate that NTON is the most potent in killing flukes in vitro among the tested flukicides and suggest the presence of TCBZ-resistant fluke populations in Bangladesh. It is imperative to explore the in vivo effects of these flukicides and subsequently their molecular mechanisms.

Control of liver fluke infections remains a significant challenge in the livestock sector due to widespread distribution of drug resistant parasite populations. In particular, increasing prevalence and economic losses due to infection with Fasciola hepatica is a direct result of drug resistance to the gold standard flukicide, triclabendazole. Sustainable control of this significant zoonotic pathogen, therefore, urgently requires the identification of new anthelmintics. Plants represent a source of molecules with potential flukicidal effects and, amongst their secondary metabolites, the diterpenoid abietic acids can be isolated in large quantities. In this study, nineteen (19) chemically modified abietic acid analogues (MC&#95;X) were first evaluated for their anthelmintic activities against F. hepatica newly excysted juveniles (NEJs, from the laboratory-derived Italian strain); from this, 6 analogues were secondly evaluated for their anthelmintic activities against adult wild strain flukes. One analogue, MC010, was progressed further against 8-week immature- and 12-week mature Italian strain flukes. Here, MC010 demonstrated moderate activity against both of these intra-mammalian fluke stages (with an adult fluke EC 50 = 12.97 µM at 72 h post culture). Overt mammalian cell toxicity of MC010 was inferred from the Madin-Darby bovine kidney (MDBK) cell line (CC 50 = 17.52 µM at 24 h post culture) and demonstrated that medicinal chemistry improvements are necessary before abietic acid analogues could be considered as potential anthelmintics against liver fluke pathogens., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Purpose: In this study, we aimed to evaluate the computed tomography (CT) findings of Fasciola hepatica infestations on initial and follow-up imagings after treatment, and also to describe the role of CT during diagnosis., Methods: In this retrospective study, patients with a diagnosis of fascioliasis by clinical and laboratory examination who underwent initial and follow-up contrast-enhanced abdominal CT scans after treatment (a single oral dose of 10mg/kg Triclabendazole) were included. The CT scans were evaluated regarding liver and spleen sizes, portal and splenic vein diameters, involved hepatic segment numbers and involvement patterns, the presence of focal perihepatic hyperdensity, gallbladder wall thickening, dilatation of the intrahepatic biliary tract, periportal-right subdiaphragmatic lymphadenopathy, hepatic subcapsular and intra-abdominal bleeding, and perihepatic/intraabdominal free fluid. Initial (pre-treatment) and post-treatment CT scans (average 25 months after the treatment) were compared with for these features., Results: A total of 36 patients with a mean age of 39.28±14.64 [15-83] years, were included in this study. After treatment, marked improvement in liver parenchymal involvement, biliary system findings, hepatomegaly-splenomegaly, periportal-peridiaphragmatic lymphadenopathy, and hepatic subcapsular hematoma were detected and focal perihepatic hyperdensity, free intraperitoneal fluid disappeared., Conclusion: Contrast-enhanced abdominal CT can be used in the diagnosis and post-treatment follow-up of fascioliasis. Awareness of intrahepatic/extrahepatic lesions and all the complications of fascioliasis can greatly aid the diagnosis and also evaluation of the response to treatment., (Copyright © 2021 Elsevier España, S.L.U. All rights reserved.)

Background: Clonorchis sinensis, Opisthorchis viverrini, and Opisthorchis felineus are the three most important human liver fluke species in the Opisthorchiidae family, infecting approximately 25 million people worldwide. Drug treatment is needed to control morbidity and is also useful in lowering transmission. Several drugs used in various regimens are available to treat these infections, but their comparative efficacy is uncertain. We aimed to compare the efficacy in terms of cure rate and egg reduction rate of currently registered drugs against human liver fluke infection., Methods: We conducted a systematic review using readily available electronic databases (MEDLINE, Embase, Cochrane Central Register of Controlled Trials, KoreaMed, China National Knowledge Infrastructure, and Wanfang Data) without language restrictions from inception until June 29, 2021. Clinical trials with pairwise comparison of drugs (praziquantel, albendazole, mebendazole, tribendimidine, or combinations of these drugs) against C sinensis, O viverrini, and O felineus were eligible, including trials comparing these drugs or their combinations with placebo. We compared efficacy in terms of cure rate by network meta-analysis. We conducted mixed binomial regression analyses for each species to derive predicted median cure rates for each drug regimen. The models included treatment and infection intensity as fixed factors, year of publication as covariate, and random effects of the different studies assumed to be normally distributed. We also assessed the quality of the included studies. This study was registered with PROSPERO (CRD42018109232)., Findings: Overall, 26 trials from 25 studies were included, of which 18 involved C sinensis, seven studied O viverrini, and one focused on O felineus. These trials included a total of 3340 participants. The two long-term treatment courses against C sinensis infection using 400 mg of albendazole (400 mg twice a day for 5 days and 400 mg twice a day for 7 days) resulted in cure rates of 100%, while two other multiple-dose regimens of albendazole resulted in high predicted cure rates: 300 mg twice a day for 5 days (93·9% [95% CI 49·6-99·6]) and 400 mg twice a day for 3 days (91·0% [50·9-99·0]). The WHO-recommended praziquantel regimen (25 mg/kg three times a day for 2 days) also showed a high predicted cure rate (98·5% [85·4-99·9]) in C sinensis infection, and predicted cure rates were above 90% for several other multiple-dose praziquantel regimens, including 20 mg/kg three times a day for 3 days (97·6% [74·7-99·8]), 14 mg/kg three times a day for 5 days (93·9% [44·8-99·7]), and 20 mg/kg twice a day for 3 days (91·0% [50·9-99·0]). In O viverrini infection, the regimen of 50 mg/kg and 25 mg/kg of praziquantel given in a single day showed the highest predicted cure rate (93·8% [85·7-97·5]), while a single dose of 50 mg/kg praziquantel also resulted in a high predicted cure rate (92·1% [64·9-98·6]). The single dose of 400 mg tribendimidine showed a high predicted cure rate of 89·8% (77·5-95·8). A low quality of evidence was demonstrated in most studies, especially those published before 2000. Selection bias due to poor random sequence generation and allocation concealment was high, and performance and detection biases were frequently unreported., Interpretation: Praziquantel shows high efficacy against clonorchiasis and opisthorchiasis. Tribendimidine might serve as a treatment alternative and warrants further investigation. Although albendazole is efficacious when long treatment schedules (5 days or 7 days) are applied, limited size of studies and high risk of bias affect the interpretation of results. More high-quality studies are needed to promote the establishment of treatment guidelines for human liver fluke infection., Funding: Fourth Round of Three-Year Public Health Action Plan (2015-2017; Shanghai, China) and Swiss National Science Foundation., Competing Interests: Declaration of interests We declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 license. Published by Elsevier Ltd.. All rights reserved.)

Fasciola hepatica, a worldwide-distributed liver fluke, is one of the causative agents of fasciolosis, a zoonotic disease that affects livestock and humans. In livestock, fasciolosis causes huge economic losses worldwide, reducing animal fertility, milk production, weight gain and condemnation of livers. In spite of the availability of drugs, such as triclabendazole (TCZ), for the treatment of fasciolosis, they do not necessarily prevent liver damage or parasite reinfection and can eventually increase parasite resistance. The aim of this research was to relate the hepatic function, haematological parameters, leukocyte counts in circulation and parasite egg shedding during F. hepatica acute and chronic phases of infection in cattle as well as to determine how these parameters change with TCZ-treatment of chronically infected cattle. Our results show that increased levels of serum aspartate aminotransferase (AST) and gamma glutamyltransferase (GGT) were detected in early stages of the experimental infection. Moreover, high circulating eosinophil count and plateletcrit levels were correlated with fluke number in livers from infected cattle. On the other hand, although TCZ-treatment in the chronic phase of infection reduced parasite burden and damage in the liver, it was not able to completely avoid them. In conclusion, our work sheds light into the physiopathological mechanisms induced during fluke infection in cattle, revealing the complexity of the host response to the infection, together with the effects of TCZ-treatment in chronically infected animals., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Fascioliasis has been reported from many countries across the globe. Acute phase of liver fluke is characterised by fever, abdominal pain and peripheral blood eosinophilia while the chronic phase is marked by chronic inflammation involving biliary tract. It is commonly diagnosed by a radiologist due to characteristic branching abscesses seen on CT scan, highly suggestive of liver fluke. Diagnosis is made by demonstration of eggs on direct microscopic examination of duodenal aspirate or bile. Screening of family members is recommended once diagnosis is has been made. Triclabendazole is the drug of choice for treatment., (Copyright © 2022 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Background: There are few reports of imported fascioliasis in Spain. This study aimed to describe the characteristics of cases registered in +REDIVI network., Methods: Observational, retrospective, descriptive study of imported fascioliasis cases registered in the +REDIVI, a multicenter collaborative network collecting information on imported infectious diseases in Spain, from October 2009 to May 2019., Results: Of 25,203 cases of imported disease registered over the study period, 16 (0.063%) were fascioliasis, acquired mainly in Pakistan, Morocco, Bolivia, and Peru. Clinical, analytical, and therapeutic data were available for 12 cases (6 immigrants, 4 people visiting friends and relatives, 2 travelers). Eleven (91.6%) had eosinophilia. The most frequent symptoms were abdominal pain (n = 5) and cough (n = 5). Two cases (16.66%) were acute and 10 (83.33%) chronic. Two patients presented lung involvement, and four had other parasitic co-infections. Twelve cases (100%) were seropositive for Fasciola hepatica. Ten patients underwent a coproparasitological study, none of which detected Fasciola spp. eggs. The probable food origin (watercress) was confirmed in 3 cases (25%). Nine of the 10 patients treated with triclabendazole (90%) and one patient treated with praziquantel were considered to meet the criteria for cure. One patient was lost to follow-up., Conclusions: Fascioliasis is a rare imported parasitosis in Spain. Eosinophilia, along with geographical origin, is the main clue for diagnosis., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

A survey of livestock producers (graziers) located in north eastern NSW Australia, collected information on perceptions and management practices for liver fluke control in livestock. The total area farmed by the 161 respondents was 195,600 ha (ranging in size from 4 to 10,522 ha) with cattle and sheep being the dominant livestock enterprises. Overall, 80% of graziers relied exclusively on anthelmintics for liver fluke control and few of these graziers (9%) integrated parasite management (IPM) strategies to reduce disease prevalence. Of those relying on anthelmintic control, triclabendazole (TCBZ) was preferentially chosen by 75% of graziers. Fifty five percent of these graziers used TCBZ in combination with oxfendazole (46%), ivermectin (5%) or abamectin (4%) whilst 45% used TCBZ as a single active ingredient. Thirty eight percent of graziers drenched livestock one or more times per year for liver fluke despite claiming they had no liver fluke or confirmed knowledge of infection. Fifty one percent of graziers based anthelmintic dose on the known weight of the heaviest animal in the herd whilst 43% visually guessed livestock bodyweight to calculate anthelmintic dose. Choice of anthelmintic was predominately based on perceived efficacy (45%) despite very few graziers (2%) having conducted post-treatment fluke egg counts. The majority of graziers (76%) were unsure if they had anthelmintic resistance, 21% claimed they had no resistance whilst 3% of graziers had confirmed resistance. Most graziers (97%) also reported farms were cohabited by kangaroos highlighting additional grazing pressures on-farm. This current survey has revealed that graziers rely on anthelmintics as their primary choice for liver fluke control. Reluctance to adopt IPM strategies and a continued heavy reliance on TCBZ, whilst basing anthelmintic decisions on perception rather than measurement and testing, pose threats for the future control of liver fluke in livestock within this endemic area., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Understanding drug penetration, distribution, and metabolization is fundamental for understanding drug efficacy. This also accounts for parasites during antiparasitic treatment. Recently, we established matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) in blood flukes and liver flukes. This label-free technique is capable of visualizing the molecular distribution of endogenous and exogenous molecules, such as drug compounds. Here, we conducted atmospheric-pressure scanning microprobe MALDI MSI (AP-SMALDI MSI) of tissue sections of adult Fasciola hepatica that have been treated in vitro with 100 µM of triclabendazole (TCBZ), the drug of choice for treatment of fasciolosis, and its main metabolite triclabendazole sulfoxide (TCBZ-SO). Measurements covered an m/z mass range of 250-1,000 and provided a high spatial resolution using a pixel size of 10 µm. To support the interpretation of drug distribution, we first identified endogenous lipids that mark characteristic tissues such as the gastrodermis, the tegument, and the parenchyma. The obtained results suggested an early tegumental route of TCBZ uptake within 20 min, followed by spreading throughout the parasite after 4 h, and an even distribution in most tissues after 12 h. This coincided with a strong reduction of parasite vitality. TCBZ-SO treatment demonstrated the accumulation of this metabolite in the same tissues as the parent drug compound. These data demonstrate the auspicious potential of MALDI MSI to visualize uptake and distribution patterns of drugs or drug-candidate compounds in parasites, which might contribute to preclinical drug discovery in liver fluke research and beyond., (© 2022. The Author(s).)

The aim of the present study was to determine the effective dose of fosfatriclaben through a field study in sheep naturally infected with F. hepatica. Thirty crossbred sheep positive for fluke eggs were selected for inclusion in the trial. On day 0, 5 groups of 6 animals each were formed for treatments: group 1 (G1), G2, and G3 received fosfatriclaben at 4, 6, and 8 mg/kg/IM, respectively. G4 received triclabendazole at 10 mg/kg/PO, and G5 was the untreated control group. Fecal samples of the sheep were analyzed to count the number of fluke eggs to evaluate the percentage of egg reduction. Twenty-one days after treatment, all sheep were humanely euthanized to extract the flukes from the bile ducts. They were counted to assess the percentage of fluke reduction. Fosfatriclaben reduced fluke eggs by 99.6, 99.6, and 100% and flukes by 94.3, 100, and 100%, respectively. Triclabendazole reduced fluke eggs by 95.2% and flukes by 100%. The fosfatriclaben injectable prodrug showed a high fasciolicidal efficacy similar to triclabendazole, with advantages over its predecessor, since only half the dose as compared to triclabendazole was required to remove eggs and flukes in the sheep that were studied., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Australian livestock are challenged by liver fluke (Fasciola hepatica) in grazing regions endemic to the intermediate snail host. Liver fluke infests a wide range of herbivores including free-roaming wildlife such as kangaroos (Macropods). The role played by Macropods in cross-species transmission and as vectors for anthelmintic resistance is largely unknown. In Phase 1 of this study, liver fluke of Eastern grey kangaroo (Macropus giganteus Shaw, 1790) origin (Kangaroo isolate) were artificially infected in sheep to confirm establishment and cross-species transmission. In Phase 2, the efficacy of triclabendazole (TCBZ) was assessed in vivo against the Kangaroo isolate to identify any drug resistance. Forty (40) merino sheep were housed in pens and allocated to one of 4 groups (Groups 1-4). Groups 1 and 2 were artificially infected with a TCBZ resistant liver fluke isolate (Oberon) originating from sheep whilst Groups 3 and 4 were infected with the Kangaroo isolate (Phase 1). At 9 weeks post infection (wpi), sheep in Groups 2 and 4 were treated with 10 mg/kg TCBZ (Phase 2). Sheep were subsequently euthanased at 11 wpi to conduct total fluke counts (TFC) in the liver. Faecal samples were collected fortnightly to measure fluke egg counts and coproantigens. Individual blood samples were collected, concurrently with faecal sampling, to monitor haematocrit and plasma proteins levels. Liver fluke of kangaroo origin established to patent infections in sheep with similar establishment and pathogenicity to the Oberon isolate. TCBZ achieved an 86 % reduction in TFC (99.8 % - adult fluke, 0 % - immature fluke) in sheep with the Kangaroo isolate and a 28 % reduction in the Oberon isolate (37 % - adult, 0 % - immature fluke). An 89 % reduction in faecal coproantigens was observed in sheep with the Kangaroo isolate and no reduction in sheep with Oberon. This study confirmed cross-species transmission of liver fluke from a kangaroo to sheep. When cohabiting the livestock grazing environment, kangaroos may act as reservoirs for liver fluke and vectors for drug resistance within liver fluke endemic areas., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The objective of this study was to evaluate the effectiveness of the main anthelmintics used for the control of trematodes in cattle in an endemic area in the humid Mexican tropics. A diagnostic study was carried out in nine cattle farms to determine the prevalence of flukes through faecal examination. Only three farms with more than 20 cows positive to trematodes were chosen to determine the effectiveness of commercial anthelmintics (triclabendazole, TCBZ; ivermectin + closantel, (IVM + CLOS); IVM + clorsulon, (CLORS); nitroxynil, NITROX). The prevalence of Fasciola hepatica was 27.1% and 29.6% of paramphistomes. The faecal egg count of trematodes ranged from 0.0 to 12.2 eggs per gram of faeces. The highest effectiveness against F. hepatica was 96.7%, and 92.7% against paramphistomes. NITROX was the most effective in the control of trematodes, while other products, such as IVM + CLORS and TCBZ obtained values lower than 90%, which puts sustainable trematode control at risk. The presence of trematodes was observed on most farms, although the prevalence per herd was highly variable, which indicates that the trematodes F. hepatica and paramphistomes are endemic to the region and a suitable management programme is suggested to control infections caused by these parasites.

Triclabendazole is the anthelminthic of choice for the treatment of fascioliasis, however, it is only registered in a few countries. We investigated the efficacy of a single-dose of triclabendazole (12 mg/kg) or albendazole (15 mg/kg) against Fasciola spp. infection in cattle on farms in the northern part of Côte d'Ivoire in a randomized clinical trial. Faecal samples were obtained from 196 cattle, of which 155 (79.1%) were found positive for Fasciola spp. by the sedimentation technique. Cattle infected with Fasciola spp. were randomly allocated (3:3:1) to receive triclabendazole (n = 66), albendazole (n = 67) or left untreated to serve as control (n = 22). Follow-up faecal samples were collected on days 21, 28, 90 and 188 post-treatment. No adverse events were observed as reported by farmers in any of the treatment groups. The proportion of non-egg shedding cattle, assessed at day 21 (primary outcome), was significantly higher in cattle treated with triclabendazole (95.4%) compared to those receiving albendazole (70.3%; odds ratio [OR] 8.73, 95% confidence interval [CI] 2.43-31.28, p <0.001). The egg reduction rate (ERR) expressed as number of eggs per gram of faeces, a secondary endpoint assessed at day 21 post-treatment, was significantly higher in the triclabendazole arm (arithmetic mean (AM) ERR = 99.8%) than in the albendazole arm (AM ERR = 92.2%), with a difference of 7.6%-points (95% CI: 0.9-14.5%-points, p=0.026). This is the first report of efficacy of triclabendazole against Fasciola spp. in naturally infected cattle in Côte d'Ivoire. Our results confirm that triclabendazole is the most effective treatment of fascioliasis and therefore, should be considered for the control of livestock fascioliasis; if resources allow in combination with intermediate host snail control and raising farmers awareness of pasture and livestock management to avoid reinfection., (Copyright © 2021. Published by Elsevier B.V.)

In this work, we present an evaluation of the fasciolicidal efficacy of a new injectable formulation of fosfatriclaben in comparison with the subcutaneous closantel and oral triclabendazole formulations currently used in veterinary practice as fasciolicides. The study was carried out in vivo on Fasciola hepatica at 2, 4, 6 and 8 weeks of age in experimentally infected sheep. To evaluate the formulation, the percent reduction of the parasite load was measured and the number of fluke eggs. Fosfatriclaben was used at 6 mg/kg/IM (dose equivalent to triclabendazole content), closantel at 5% at 10 mg/kg/SC, and triclabendazole at 10 mg/kg/PO; the control group received no treatment. Fosfatriclaben showed fasciolicidal efficacies of 95.5 %, 100 %, 100 % and 100 %, and triclabendazole showed similar efficacies of 97.4 %, 100 %, 100 % and 100 %, at the different treatment weeks (P > 0.05). Closantel showed limited efficacy against 2-, 4- and 6-week-old flukes but 100 % efficacy in adult flukes. All three evaluated formulations eliminated all 8-week-old F. hepatica trematode eggs. Although fosfatriclaben and triclabendazole showed similar fasciolicidal efficacy, the intramuscular administration of fosfatriclaben has several advantages over the oral administration of triclabendazole, such as ease of administration for veterinary use and a reduced risk of accidents for both the operator and the animals. In addition, the dose used in this injectable formulation is only 60 % of the oral dose, which reduces environmental contamination., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Fascioliasis is a foodborne zoonotic infection caused by the trematode liver flukes: Fasciola hepatica and Fasciola gigantica (F. gigantica). Infections may cause acute and chronic hepatobiliary tract diseases in herbivore animals and humans. Fascioliasis is present worldwide, particularly in regions where sheep and cattle are raised. The global burden of human fascioliasis is estimated to be 2.7 million. Human infections are rare in the United States, and most infections were acquired abroad. In the 1950s, several human cases of F. gigantica infection were reported from Hawai'i, but no subsequent cases have been reported until the case described here. This case report describes a man from Hawai'i Island who ate raw wild watercress, and 22 days later, developed acute phase fascioliasis with fever, acute hepatitis, peripheral eosinophilia, and hypodense lesions seen on liver diagnostic imaging. Immunodiagnostic tests were positive for Fasciola species. Based on earlier reports from Hawai'i, F. gigantica infection was likely. Clinical abnormalities resolved after treatment with triclabendazole. Physicians should consider fascioliasis in immigrants and travelers from endemic areas with acute hepatitis and eosinophilia after eating raw wild watercress or chronic hepatobiliary disease. Hepatobiliary imaging and serological testing are useful for diagnosis. Oral triclabendazole is the preferred treatment. Animal fascioliasis appears to be spreading in the United States, and the popularly perceived health benefits of eating raw wild watercress and other aquatic plants may lead to more human infections. The rarity of human infections in Hawai'i suggests that it is safe to eat commercially grown watercress cultivated in Hawai'i., (©Copyright 2021 by University Health Partners of Hawai‘i (UHP Hawai‘i).)

We report a case of human fascioliasis in the USA that encountered many diagnostic uncertainties. Numerous tests available for detection of fascioliasis were utilised but the diagnosis remained elusive. Confounders included three negative stool ova and parasite examinations, positive hepatitis A virus IgM antibody, cross-reactive false-positive Echinococcus IgG antibody, absence of characteristic image findings and unrevealing liver biopsy. Praziquantel was started as empiric treatment for helminth infections, but was ineffective. Due to the rarity of the disease in the USA, serologic testing and triclabendazole were only available from the Centers for Disease Control and Prevention, which led to a delay in diagnosis and treatment., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

Introduction: The only drug effective against the infection caused by Fasciola hepatica or F. gigantica is triclabendazole (TCBZ), recommended by the WHO and recently approved by the FDA. Here, we describe the evolution of TCBZ regimens and the emergence of TCBZ failure to Fasciola infection., Areas Covered: The present review focuses on the evidence of TCBZ for the treatment of fascioliasis. For acute fascioliasis, there is a lack of studies to measure the presence of eggs of Fasciola in stool samples on the follow-up after initial TCBZ treatment. For chronic fascioliasis, WHO recommends a single oral dose of TCBZ 10 mg/kg whereas CDC recommends two doses of TCBZ 10 mg/kg 12 h apart. Incremental number of treatment failures have been documented worldwide. There are currently no therapeutic alternatives for the treatment of fascioliasis in humans., Expert Opinion: Most cases of human fascioliasis are successfully treated with TCBZ, but some continue excreting eggs in the stools despite 1-2 standard of care regimens of TCBZ. A precise regimen is unclear for those patients who fail the initial treatment with TCBZ. Further clinical trials are needed to address the possible TCBZ emerging resistance.

Background: Fascioliasis is a food-borne hepatobiliary zoonosis caused by Fasciola hepatica and Fasciola gigantica. Human infestations are predominantly seen in developing countries where the disease is endemic, but, due to the increase in international travel rates, hepatic fascioliasis is also appearing in nonendemic areas including Europe and the USA. The clinical and laboratory findings are usually nonspecific. Cross-sectional imaging can be very helpful in the diagnosis of fascioliasis as well as to differentiate it from other liver diseases with a very similar clinical picture. The objectives of this case report are to discuss imaging findings of hepatic fascioliasis and to review the literature., Case Presentation: We report the case of a 35-year-old Iranian patient who presented with right upper quadrant pain, low-grade fever, fatigue, and anorexia. The patient had a history of recent travel to the Gilan Province of Iran, almost a month before the onset of symptoms, which is an endemic area of fascioliasis in the country. Laboratory examinations revealed eosinophilia, elevated hepatic enzymes, and slightly raised C-reactive protein. Contrast-enhanced computed tomography of the patient shows clusters of focal ill-defined hypodense lesions with mild peripheral enhancement in the right liver lobe and subcapsular regions. Magnetic resonance imaging of the liver revealed multiple ill-defined lesions of low signal intensity on the T1-weighted image and high signal intensity on the T2-weighted image, extending from the liver capsule into deeper parenchyma toward periportal regions, which shows mild peripheral enhancement on post-contrast images. Imaging-based diagnosis of fascioliasis was made depending on the characteristic distribution of subcapsular tracts/lesions on the above-mentioned imaging, which was then confirmed by serologic tests using enzyme-linked immunosorbent assay. The patient was treated with triclabendazole, showing great clinical improvement, and was eventually discharged in good health condition., Conclusion: The imaging findings in this case report highlight the importance of cross-sectional imaging for further evaluation of suspected cases of fluke-induced liver disease. The hypothesis of hepatic fascioliasis should be always considered when consistent radiological findings are observed. Clusters of tortuous subcapsular lesions with peripheral contrast enhancement extending into deeper liver parenchyma are characteristic imaging findings that strongly suggest hepatic fascioliasis.

Background: The popularity of new world camelids, particularly alpacas, is growing rapidly in Ireland, presenting a clinical challenge to veterinary practitioners who may not have worked with these species previously. To the authors' knowledge, the clinical course of a case of acute fasciolosis in an alpaca has not previously been reported, and fasciolosis has not been reported at all in alpacas in Ireland, making this case report a valuable addition to the current literature., Case Presentation: A three-year-old male castrated huacaya alpaca was admitted to UCD Veterinary Hospital with a two-day history of colic and tenesmus. He had been treated with albendazole, dexamethasone and potentiated amoxycillin by the referring veterinary practitioner with no response. On initial clinical exam, sensitivity to abdominal palpation was the only abnormality. However, the alpaca proceeded to show abnormal lying positions, tenesmus and reduced faecal output over the next 24 h. A general blood panel demonstrated moderate anaemia, marked hyperglobulinaemia and moderately increased hepatocellular and hepatobiliary enzyme activity. Abdominal radiography revealed enlargement of the first forestomach compartment without evidence of gastrointestinal obstruction or peritonitis. An abdominal ultrasound exam revealed an elongated, heterogenous mass in the caudoventral abdomen that appeared to be contiguous with the liver. FNA of this mass revealed that it was in fact a liver lobe with biliary stasis and inflammation. Faecal sedimentation demonstrated Fasciola hepatica eggs. In spite of treatment with triclabendazole and supportive treatment including blood transfusion, the alpaca's condition continued to deteriorate and he was euthanised. On post-mortem exam, acute fasciolosis was diagnosed., Conclusions: The clinical presentation and course of a case of acute fasciolosis in an individual alpaca is described, including the results of a range of diagnostic tests that were carried out. The final diagnosis is supported by a description of post-mortem findings. This information will serve as a resource for veterinary practitioners involved in the diagnosis and treatment of similar cases.

Background: Human fascioliasis, caused by the liver flukes F. hepatica, and F. gigantica, is a neglected tropical disease that causes health problems in many regions of the world. This disease can be classified as either acute or chronic based depending on the clinical manifestations and laboratory findings., Methods: We retrospectively reviewed the demographic data, clinical features, radiologic manifestations, and the response to specific treatment of patients diagnosed with hepatic fascioliasis as well as fasciola liver abscess in Thailand., Results: A total of 175 patients were included in the study, 126 patients were females (72%), while the mean age was 47.8 years (16-84 years). The most common symptoms were abdominal pain (74.9%), weight loss (29.1%) and fever (28%). Peripheral eosinophilia was observed in 92% of patients. The typical radiologic findings discovered conglomerated hypodensity which are rim-enhancing lesions located in the subcapsular and peripheral region of the liver. Most of patients were improved after a single dose of triclabendazole treatment. Adding antibiotic had no statistical impact on treatment outcome (p = 0.78)., Conclusions: Human fascioliasis presents with a wide clinical spectrum; therefore, a high index of suspicion is required to establish a correct diagnosis. Clinicians need to be aware of hepatic fascioliasis when patients in such endemic areas present as hypereosinophilia and typical liver imaging. Prompt specific treatments will contribute towards a satisfactory outcome in patients., (© 2021. Asian Pacific Association for the Study of the Liver.)

The use of natural products for disease control is a promising approach to solving the problem of drug resistance. The aim of the research reported here was to evaluate the fasciolicidal and anti-Clostridium novyi type B activities of propolis administered orally to sheep infected with Fasciola gigantica and C. novyi type B. Sheep infected with both pathogens were divided into two groups: an infected treated group and an infected non-treated group. The treatment was oral administration of 50 mg propolis extract/kg daily for 15 days. The body weight of the sheep, fecal egg counts of F. gigantica, serum levels of F. gigantica IgG, concentrations of cytokines (IL-2, IL-10, and IL-17), and bacterial counts of C. novyi were evaluated. Following treatment, the sheep had increased body weight and a significant decrease in the egg count, which was reduced by 54.54% at 15 days post treatment. The level of anti- Fasciola IgG increased, whereas levels of IL-2, IL-10, and IL-17 decreased in propolistreated sheep. Treatment of sheep with propolis produced a significant reduction in fecal count of C. novyi, from 8 × 10 9 to 3 × 10 3 colony units per gram at 15 days post treatment. This research highlights the therapeutic potential of Egyptian propolis extract as a treatment against F. gigantica and C. novyi type B infections, and investigated its mode of action through its effect on some cellular and humoral responses in sheep with both infections.

Objectives: Triclabendazole (TCBZ) is the recommended anthelmintic against Fasciola hepatica at a dose of 10 mg/kg body weight administered as a single or double dose. However, treatment failures to TCBZ standard-of-care (SOC) doses have been reported in humans. The aim of this study was to describe the effectiveness and tolerability of alternative TCBZ regimens in those patients who failed the SOC regimen for fascioliasis in Peru., Methods: A retrospective study was conducted at a major referral centre for fascioliasis in Peru between 2002 and 2018. Inclusion criteria were cases with chronic F. hepatica infection who failed the SOC regimen for human fascioliasis with TCBZ at 10 mg/kg orally as single dose with food., Results: A total of 27 cases (59% female; mean age 39.4 years, range 6-71 years) with chronic fascioliasis failed at least once the current SOC regimen of TCBZ. Of 27 cases, 21 failed a second treatment regimen. Multiple regimens of TCBZ were given to these patients until three consecutive stool examinations were negative for Fasciola eggs. The overall cure rate was 74%. TCBZ was well tolerated with minimal side effects., Conclusion: According to the results of this study, patients labelled as having 'TCBZ resistance' may respond to multiple regimens of TCBZ with a cure rate of 74%. Thus, the term 'TCBZ resistance' should be re-evaluated using biomarkers., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Pharmacological treatment remains essential to control fasciolosis in areas where infection is endemic. However, there are major constraints to treating food-producing animals. Of particular concern is the lack of flukicides for treating early Fasciola infections in ruminant livestock in some countries. In addition, the information provided in package leaflets, particularly regarding withdrawal periods, is often incomplete, confusing, and/or contradictory. International regulatory bodies should harmonize the use of flukicides in livestock in favor of fairer, safer international trade. In addition, monitoring the efficacy of fasciolicides on farms is also essential to minimize the spread of drug-resistant populations of Fasciola. The current situation regarding flukicide formulations in the European Union and other, non-European countries is analyzed in this review paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)