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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. [ABSTRACT FROM AUTHOR]

The objective of the present study was to describe a very rare case of gallstone (cholelithiasis) in a goat associated with chronic fasciolosis. During a routine slaughterhouse‐based survey, a two‐and‐half‐year‐old female Black Bengal Goat was found to be affected with severe chronic fascioliosis characterized by the massive damage in the liver. Through systemic dissection of liver, we isolated 94 adult Fasciola spp., and by PCR, we confirmed the fluke as Fasciola gigantica. The gallbladder of the goat was oedematous. On opening the gallbladder, we recovered 255 stones of variable sizes. Stones were whitish in colour and friable, and some of the fragile stones were attached to the wall of the gallbladder. To the authors' knowledge, this is the first report of the cholelithiasis in a goat associated with F. gigantica. [ABSTRACT FROM AUTHOR]

Background: The insulin/insulin-like signalling (IIS) pathway is common in mammals and invertebrates, and the IIS pathway is unknown in Fasciola gigantica. In the present study, the IIS pathway was reconstructed in F. gigantica. We defined the components involved in the IIS pathway and investigated the transcription profiles of these genes for all developmental stages of F. gigantica. In addition, the presence of these components in excretory and secretory products (ESPs) was predicted via signal peptide annotation. Results: The core components of the IIS pathway were detected in F. gigantica. Among these proteins, one ligand (FgILP) and one insulin-like molecule binding protein (FgIGFBP) were analysed. Interestingly, three receptors (FgIR-1/FgIR-2/FgIR-3) were detected, and a novel receptor, FgIR-3, was screened, suggesting novel functions. Fg14-3-3ζ, Fgirs, and Fgpp2a exhibited increased transcription in 42-day-old juveniles and 70-day-old juveniles, while Fgilp, Fgigfb, Fgsgk-1, Fgakt-1, Fgir-3, Fgpten, and Fgaap-1 exhibited increased transcription in metacercariae. FgILP, FgIGFBP, FgIR-2, FgIR-3, and two transcription factors (FgHSF-1 and FgSKN-1) were predicted to be present in FgESPs, indicating their exogenous roles. Conclusions: This study helps to elucidate the signal transduction pathway of IIS in F. gigantica, which will aid in understanding the interaction between flukes and hosts, as well as in understanding fluke developmental regulation, and will also lay a foundation for further characterisation of the IIS pathways of trematodes. [ABSTRACT FROM AUTHOR]

Background: The potential molluscicidal extracts, obtained from indigenous plants Cannabis sativa, Acacia nilotica, and Tinospora cordifolia, were tested for toxicity against freshwater pulmonate snail Lymnaea acuminata, an intermediate host of Fasciola hepatica. The organic extracts had a significant effect on young snails. Materials and Methods: All organic extracts and column-purified fractions gave median lethal concentrations (19–100.05 mg/L; 24 h) that fell well within the threshold level of 100 mg/L, set for a potential molluscicide by the World Health Organization. Results: The toxicity of T. cordifolia stem acetone extract (96 h LC50: 16.08 mg/L) was more pronounced compared with C. sativa leaf ethanol extract (96 h LC50: 16.32 mg/L) and A. nilotica leaf ethanol extract (96 h LC50: 24.78 mg/L). β-caryophyllene, gallic acid, and berberine were characterized and identified as active molluscicidal components. Co-migration of β-caryophyllene (retardation factor [Rf] 0.95), gallic acid (Rf 0.30), and berberine (Rf 0.23) with column-purified parts of Cannabis sativa, Acacia nilotica, and Tinospora cordifolia on thin-layer chromatography demonstrates same Rf value, that is, 0.95, 0.30, and 0.23, respectively. Conclusion: This study indicates that these extracts thus represent potential plant-derived molluscicides that are worthy of further investigations. [ABSTRACT FROM AUTHOR]

Background: Snails of the Lymnaeidae family are the intermediate hosts of Fasciola species, the causative agents of fascioliasis. The purpose of this study was to determine the prevalence of Fasciola species in lymnaeid snails and to investigate the association of geoclimatic factors and Fasciola species distribution in northwestern provinces of Iran using geographical information system (GIS) data. Methods: A total of 2000 lymnaeid snails were collected from 33 permanent and seasonal habitats in northwestern Iran during the period from June to November 2021. After identification by standard morphological keys, they were subjected to shedding and crushing methods. Different stages of Fasciola obtained from these snails were subjected to the ITS1 polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method for species identification. The associations of weather temperature, rainfall, humidity, evaporation, air pressure, wind speed, elevation, and land cover with the distribution of Fasciola species were investigated. Geographical and statistical analysis was performed using ArcMap and SPSS software, respectively, to determine factors related to Fasciola species distribution. Results: Of the 2000 snails collected, 19 were infected with Fasciola hepatica (0.09%), six with F. gigantica (0.03%), and 13 with other trematodes. Among geoclimatic and environmental factors, mean humidity, maximum humidity, and wind speed were significantly higher in areas where F. hepatica was more common than F. gigantica. The altitude of F. hepatica-prevalent areas was generally lower than F. gigantica areas. No significant relationship was observed between other investigated geoclimatic factors and the distribution of infected snails. Conclusions: The present study showed the relationship of humidity and wind speed with the distribution of snails infected with F. hepatica or F. gigantica in the northwestern regions of Iran. In contrast to F. gigantica, F. hepatica was more prevalent in low-altitude areas. Further research is recommended to elucidate the relationship between geoclimatic factors and the presence of intermediate hosts of the two Fasciola species. [ABSTRACT FROM AUTHOR]

The occurrence of trematodes among ruminants and their snail vectors is a major concern across various agro-ecological regions of Ethiopia. Trematodes pose significant threats to animals, causing considerable economic losses and impacting public health. In this study, we have investigated 784 ruminant fecal samples, and 520 abattoir samples, alongside the collection and identification of snail vectors from various agro-ecological regions. Fecal examinations revealed Fasciola, Paramphistomum and Schistosoma species infected 20.5% (95% CI: 17.6, 23.8), 11.7% (95% CI: 9.6, 14.2), and 6.3% (95% CI: 4.1, 9.1) of the animals, respectively. The overall prevalence of trematodes among ruminants was 28.8% (95% CI: 25.7, 32.1%), with 6.0% (95% CI: 4.3, 7.7) showing mixed infections. Fasciola was more prevalent in Asela (26%) compared to Batu (19%) and Hawassa (11.5%), while a higher proportion of animals in Batu were infected with Paramphistomum. Schistosoma eggs were detected only in Batu (12.5%), but not in other areas. Sheep and cattle exhibited higher infection rates with Fasciola, Paramphistoma, and Schistosoma compared to goats. Significant associations were observed between trematode infections and risk factors including agro-ecology, animal species, body condition score, and deworming practices. About 20.8% and 22.7% of the slaughtered animals harbored Fasciola and Paramphistomum flukes, respectively, with a higher prevalence in Asela and Hawassa abattoirs compared to Batu abattoir. Additionally, a total of 278 snails were collected from the study areas and identified as lymnae natalensis, lymnae trancatula, Biomphalaria pffiferi, Biomphlaria sudanica, and Bulinus globosus. In conclusion, the study highlights the widespread occurrence of trematode infections, emphasizing the need for feasible control measures to mitigate their economic and public health impacts. [ABSTRACT FROM AUTHOR]

Fascioliasis and toxoplasmosis are the two important zoonotic diseases that are endemic in Iran and share some common transmission routes. The present study is aimed at determining the seroprevalence of human fascioliasis and toxoplasmosis in rural and urban areas of Jolfa County, Northwest Iran. In a cross-sectional study, 600 human sera were collected randomly from humans living in Jolfa County including three cities and 13 villages from 2017 to 2018. Anti-Toxoplasma IgG and anti-Fasciola sp. IgG tests have been performed using the enzyme-linked immunosorbent assay. Four (0.7%) out of 600 human sera showed positive levels of anti-Fasciola IgG. Three out of four seropositive humans were from an urban area, and one (25%) was from rural inhabitants. Considering T. gondii infection, 45% of studied human sera were seropositive for anti-T. gondii IgG. In conclusion, this is the first study reporting Fasciola seropositivity in the area. Based on the findings, human fascioliasis is present in the studied area, Northwest Iran, granted in low prevalence. Considering T. gondii seropositivity, the prevalence is high, yet close to the reports from other regions in the province. [ABSTRACT FROM AUTHOR]

[Display omitted] • Wider distribution of invasive snails compared with native snails in the Limpopo River system. • cox 1 genetic data shows multiple lineages of invasive snails outside their native ranges. • More digenean species occurred in Pseudosuccinea columella than in the other snails. • The present xiphidiocercaria represents an unknown species of Orientocreadium. Invasive snails are associated with ecological problems in freshwater bodies worldwide. However, their impact on the transmission of digenean infections remain underreported. In the present study, 1708 specimens representing four snail species were sampled from four impoundments in the Limpopo River system in South Africa. Gyraulus chinensis (Planorbidae), Physella acuta (Physidae) and Pseudosuccinea columella (Lymnaeidae), which are invasive, were found in all the sampling sites. In contrast, the native lymnaeid Radix natalensis occurred at only one study site. Digeneans were observed only from R. natalensis (prevalence = 49%) and Ps. columella (prevalence = 23%). Morphological and genetic analyses revealed four digeneans: Fasciola nyanzae , Orientocreadium sp., Petasiger sp. and Patagifer vioscai. Pseudosuccinea columella was infected by the four digeneans while R. natalensis harboured only Orientocreadium sp. and Petasiger sp. Partial sequences of Orientocreadium sp. from the current study differed from congeners whose DNA data are available on GenBank, by p-distances of at least 1.84 and 2.2% for 28S and the internal transcribed spacer (ITS) rDNA, respectively. Phylogenetic analyses demonstrated that the present species is sister to Orientocreadium batrachoides. Genetic and phylogenetic data based on 28S and ITS rDNA suggested that Petasiger sp. from the present study and isolates of three unidentified Petasiger spp. from Kenya, Hungary and Australia, were representatives of the same species. This is the first known report of Orientocreadium , Petasiger and Patagifer from Ps. columella. The occurrence of F. nyanzae in Ps. columella indicates spillback from R. natalensis. These findings echo the concerns raised in previous studies about the potential role of Ps. columella in the amplification of digenean diseases in its introduced range. Phylogenetic analyses of partial sequences of the cytochrome c oxidase subunit 1 mitochondrial gene (cox 1) showed multiple lineages of Ps. columella in North and South America. Pseudosuccinea columella specimens from the present study belong to an invasive genotype that has spread globally and has been reported from Zimbabwe, Egypt, Portugal, Australia, Argentina, Colombia and New Mexico (USA). Physella acuta from the current study had a stronger genetic relationship with isolates from Canada and Iceland, than with isolates from other parts of Africa, suggesting several invasion routes into Africa. This is the first known DNA characterisation of G. chinensis from Africa. Phylogenetic reconstruction indicated multiple exit events of G. chinensis from Asia into Europe and Africa. South African isolates clustered in a recent branch containing isolates from the Czech Republic and Hong Kong, China. Considering the presence of invasive snails in all the sampling sites in the present study, it is necessary to investigate the factors that enhance their establishment and to monitor their effects on the native snail populations. [ABSTRACT FROM AUTHOR]

Starting in October 2021, quarterly malacological surveys have been undertaken in Malawi, with the sampling of 12 specified freshwater habitats throughout a calendar year. Each survey monitors the presence of aquatic intermediate snail hosts of medical and veterinary importance. In March 2023, the alien lymnaeid species Pseudosuccinea columella was encountered for the first time in the surveys, in Nsanje District. This species identity was later confirmed upon DNA analysis of mitochondrial ribosomal 16S sequences. In July 2023, P. columella was also noted at single sites within Mangochi and Chikwawa Districts, and again in Nsanje District, with an additional location observed. Of particular importance, our sampled location in Mangochi District was directly connected to Lake Malawi, which expands the species list of invasive molluscs in this lake. While P. columella is a well-known intermediate snail host for human and animal fascioliasis, screening collected snails for trematode cercariae, alongside molecular xenomonitoring, did not yield equivocal evidence of active fluke infection. However, the newly recognized presence of this alien intermediate snail host within Lake Malawi, and along the Shire River Valley, flags a new concern in altered local transmission potential for human and animal fascioliasis. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The Drosophila repleta group is one of the most widely used model systems for studying adaptation and speciation. Traditionally, five subgroups are recognized within the repleta group: fasciola, hydei, mercatorum, repleta and mulleri. A sixth subgroup, inca, was the last to be defined. The inca species subgroup includes three species: Drosophila inca, Drosophila huancavilcae and Drosophila yangana, all of which are endemic to Ecuador and Peru. Previous molecular phylogenetic studies have been inconclusive regarding the geographic location, time and mode of diversification of lineages within the repleta group. By applying a phylogenetic and biogeographical analysis of 54 taxa belonging to the repleta, nannoptera, atalaia and virilis groups, the aim of this study was to: (1) determine the relationships between the inca subgroup and the other five subgroups within the repleta group, (2) improve the unresolved branching and low supports of the early divergent lineages in the repleta group phylogeny and (3) estimate the geographic and temporal context of the early divergence within the repleta group. Based on these findings, it is proposed that the Drosophila repleta group first diversified during the mid-late Miocene, most likely following the uplift of the Northwestern Andes. [ABSTRACT FROM AUTHOR]

Intrashell oxygen isotope (δ18Os) analyses of terrestrial snails have been carried out over two decades. However, the intraspecies/interspecies differences are not yet well understood. Here, we conducted a high‐resolution intrashell δ18Os study on 43 shells from Cathaica fasciola and Bradybaena ravida, and 1449 δ18Os data were obtained. These large amounts of data demonstrate the reproducibility of intrashell δ18Os. Our results suggest that C. fasciola and B. ravida have similar growing seasons mainly from March to October in the studied region. The ecological habits of the two species could explain the distinctive variations in their δ18Os sequences. Therefore, our study may interpret the discrepancy between the studies of land snail species and the stable/clumped isotopes of their shells. Moreover, the calculated average growth rate is ∼290 μm/day (from 250 to 330 μm/day) for nonadult C. fasciola. Thus, the use of snail shells for reconstructing high‐resolution terrestrial climate changes is promising. Plain Language Summary: Land snails are sensitive to climatic and environmental changes. Therefore, the oxygen isotopic composition of snail shells (δ18Os) has often been used to study environmental changes. Researchers started to collect multiple samples from a single shell for δ18Os analyses about two decades ago. However, it is still unclear whether high‐resolution intrashell δ18Os results are repeatable and whether the influences of different individuals/species may exist. To address these issues, we collected 43 snail shells from Cathaica fasciola and Bradybaena ravida. In total, 1449 δ18Os data were obtained. The large amount of data showing similar variations in the intrashell δ18Os from different shells demonstrates the repeatability. Accordingly, our data suggest that the two species have similar growing seasons, mainly from March to October, although their ecological habits are different. In addition, the ecological habits of these two species could explain the subtle differences in their δ18Os. Taken together, our study could resolve the discrepancy between previous work on land snail species and stable/clumped isotopes. Furthermore, the estimated average growth rate is ∼290 μm/day (from 250 to 330 μm/day) for nonadult C. fasciola. Overall, snail shells are promising records for exploring high‐resolution terrestrial climate changes in the future. Key Points: The intrashell δ18Os of land snails is reproducible, which mainly records the δ18O of precipitationCathaica fasciola and Bradybaena ravida have different ecological habits but similar growing seasonEcological habits could explain the discrepancy between studies of land snail species and stable/clumped isotopes [ABSTRACT FROM AUTHOR]

Radix (Lymnaea) natalensis (Krauss, 1848) is an essential host in the completion of the life cycle of Fasciola gigantica especially in large areas of Africa, where it is reputed to serve as the main, if not the only intermediate of Fasciola. In this study, 18S rDNA gene was employed for the molecular characterization of morphologically identified Radix natalensis collected from Nigerian Northern cities of Gombe and Jos. The obtained result revealed that the amplification of the 18S rDNA gene at 450bp identified and confirmed the collected species as Radix natalensis. BLASTn search showed that sequences of Nigerian isolates of Radix natalensis had high (99-100%) degree of similarities with each other and with other sequence deposits in the NCBI GenBank. The Nigerian sequences on phylogenetic analysis formed a separate clade distinct from other sequences of Radix natalensis from other parts of Africa and the rest of the world, suggestive of clustering based on geographical location. Thus, the availability and confirmation of Radix (Lymnaea) natalensis from the study locations as evidenced by the results of this study further justifies the assertion of Lymnaea natalensis, being a putative intermediate host of Fasciola spp. in Nigeria. [ABSTRACT FROM AUTHOR]

Background: Fasciola gigantica is a helminth parasite of veterinary importance and a constraint on the growth and productivity of domestic livestock in the tropical countries of Asia and Africa. Present study aimed at determining the expression of four cysteine protease genes including cathepsin (cat) L-1 D and cat-Bs in the embryonated egg stage of F. gigantica by polymerase chain reaction (PCR). In addition to the analysis of expression of the above genes in the embryonated eggs, inhibition of in vitro hatching of the miracidia from the eggs was carried out using three protease inhibitors viz. serine, cysteine and leucine aminopeptidase inhibitors to understand the possible role of these proteases in the hatching of the miracidia from the eggs. Methods: Cat- L-1D, cat B-1, cat B-2 and cat B-3 genes were amplified by reverse transcriptase and quantitative-PCR, respectively for determining their expression in the embryonated eggs. Inhibition of in vitro hatching of the miracidia from the eggs was carried out by exposing the embryonated eggs to different concentrations of iodoacetamide, phenyl methyl sulphonyl fluoride and bestatin as cysteine, serine and leucine aminopeptidase inhibitors. Result: Results showed that cat L1-D, cat B-1 and cat B-3 genes were expressed at the embryonated egg stage but cat B-2 gene was not expressed at this stage of the parasite. Iodoacetamide and phenyl methyl sulphonyl fluoride (cysteine and serine protease inhibitors) inhibited the miracidial hatching at different concentrations, indicating probable role of cysteine and serine proteases in its hatching process. However, leucine aminopeptoidase does not seem to influence the miracidial hatching from the eggs as bestatin did not inhibit its hatching. [ABSTRACT FROM AUTHOR]

Fascioliasis is a zoonotic parasitic infection caused by Fasciola species in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validating methods for the diagnosis of fascioliasis in humans. Serological techniques are convenient assays that significantly improves the diagnosis of Fasciola infection. However, a more sensitive method is required. The aim of this study was to compare the Real-Time PCR technique with the indirect-ELISA for the detection of Fasciola hepatica in human. Using a panel of sera from patients infected with Fasciola hepatica (n = 51), other parasitic infections (n = 7), and uninfected controls (n = 12), we optimized an ELISA which employs an excretory–secretory antigens from F. hepatica for the detection of human fascioliasis. After DNA extraction from the samples, molecular analysis was done using Real-Time PCR technique based on the Fasciola ribosomal ITS1 sequence. Of 70 patient serum samples, 44 (62.86%) samples were identified as positive F. hepatica infection using ELISA and Real-Time PCR assays. There was no cross-reaction with other parasitic diseases such as toxoplasmosis, leishmaniasis, taeniasis, hydatidosis, trichinosis, toxocariasis, and strongyloidiasis. The significant difference between the agreement and similarity of the results of patients with indirect ELISA and Real-Time PCR was 94.4% and 99.2%, respectively (Cohen's kappa ≥ 0.7; P = 0.02). Based on the Kappa agreement findings, the significant agreement between the results of ELISA and Real-Time PCR indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in humans. [ABSTRACT FROM AUTHOR]

Liver flukes are parasitic flatworms that can infect humans and other mammals. In this study, freshwater snails from Chaharmahal and Bakhtiari province in Iran were investigated for the presence of liver fluke larvae. The snails were collected from various rivers and analysed using standard morphological and molecular techniques. Additionally, molecular identification of the collected snails was conducted through polymerase chain reaction (PCR) and sequencing of the COI gene. The freshwater snails were identified as Physella acuta (Draparnaud, 1805), Galba truncatula (O.F. Müller, 1774), Radix euphratica (Mousson, 1874), Radix auricularia (Linnaeus, 1758) and Planorbis planorbis (Linnaeus, 1758). The presence of larval stages of Fasciola spp. in individual snails was examined using stereomicroscopy and a crushing method. Morphological and molecular identification based on the ITS-1 marker for identifying Fasciola spp. revealed that 10.9% (59 out of 549) of R. auricularia and 5.8% (37 out of 630) of P. acuta were infected. A blastn analysis of the collected samples revealed that all isolates belonged to F. hepatica, indicating a potential risk of liver fluke infection in the study area. These findings emphasise the importance of regular surveillance and control measures to prevent the spread of these parasites. [ABSTRACT FROM AUTHOR]

Background: Fasciolosis is an emerging public health threat in a number of regions worldwide. To date, we lack an overview of both its occurrence and distribution in Southeast Asia across all actors involved in the life cycle, which impedes the development of disease control measures. Therefore, our objective was to collect recent information on the distribution and the prevalence of Fasciola spp. and the associated risk factors for infection in humans, animals, snails and plant carriers in Southeast Asia. Methodology: Bibliographic and grey literature databases as well as reference lists of important review articles were searched for relevant records published between January 1st, 2000, and June 30th, 2022. The systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines for reporting systematic reviews. A total of 3,887 records were retrieved, of which 100 were included in the final analysis. Principal findings: The studies focused mainly on one host species (96.0%), with Fasciola spp. infection in animals being the most studied (72.0%), followed by humans (21.0%). Based on the used inclusion and exclusion criteria, reports were retrieved describing the presence of Fasciola spp. infection was in seven out of 11 countries in Southeast Asia. Depending on the diagnostic tool applied, the prevalence of Fasciola spp. infection ranged between 0.3% and 66.7% in humans, between 0% and 97.8% in animals, and between 0% and 66.2% in snails. There were no studies reporting the presence of metacercariae on plant carriers. Conclusions/Significance: Our study reconfirms that Fasciola spp. infections are widespread and highly prevalent in Southeast Asia, but it remains difficult to accurately assess the true occurrence of Fasciola spp. in absence of well-designed surveys covering all hosts. As next steps we propose to assess the occurrence of the infection across all actors involved in the transmission, to identify associated risk factors and to estimate the burden of the disease to support national and international decision makers. Author summary: Fasciolosis is a foodborne disease that is caused by a parasitic worm. The disease affects both public and animal health worldwide. The transmission of Fasciola is complex and involves humans, animals and plants. The adult worms are observed in both humans and ruminants, while larval stages develop in freshwater snails. The infective larval stage attaches to aquatic plants. In recent decades, human fasciolosis has been considered an emerging disease in many regions of the world, including Southeast Asia. Although various studies surveyed Fasciola infection, an overview of existing knowledge on the distribution and prevalence of Fasciola across the different hosts is missing for this region. Yet, this is important to develop appropriate disease control measures. In this study, we systematically searched for literature on Fasciola spp. infection in Southeast Asia. Our study reconfirms that Fasciola is widespread in this region, but that it remains difficult to accurately assess the occurrence of parasite in the absence of well-designed surveys. A firm response to control fasciolosis requires both transdisciplinary and international partnerships. As next steps we propose to assess the occurrence of Fasciola spp. across all actors involved in the transmission, to identify associated risk factors and to estimate the disease burden to support national and international decision makers. [ABSTRACT FROM AUTHOR]

The Drosophila repleta group is one of the most widely used model systems for studying adaptation and speciation. Traditionally, five subgroups are recognized within the repleta group: fasciola, hydei, mercatorum, repleta and mulleri. A sixth subgroup, inca, was the last to be defined. The inca species subgroup includes three species: Drosophila inca, Drosophila huancavilcae and Drosophila yangana, all of which are endemic to Ecuador and Peru. Previous molecular phylogenetic studies have been inconclusive regarding the geographic location, time and mode of diversification of lineages within the repleta group. By applying a phylogenetic and biogeographical analysis of 54 taxa belonging to the repleta, nannoptera, atalaia and virilis groups, the aim of this study was to: (1) determine the relationships between the inca subgroup and the other five subgroups within the repleta group, (2) improve the unresolved branching and low supports of the early divergent lineages in the repleta group phylogeny and (3) estimate the geographic and temporal context of the early divergence within the repleta group. Based on these findings, it is proposed that the Drosophila repleta group first diversified during the mid-late Miocene, most likely following the uplift of the Northwestern Andes. [ABSTRACT FROM AUTHOR]

The traditional diagnostic method of fascioliasis was based on the identification of the eggs in the stool, but this is not a reliable way as it has many restrictions. The present work was evaluated to immunodetect the Excretory/ Secretory antigens of Fasciola gigantica by sandwich Elisa in naturally infected cattle's sera and by dot blot in naturally infected human sera and compare these methods and the traditional methods. In this study, fresh adult Fasciola gigantica worms were collected to extract crude excretory/secretory (E/S) antigens. E/S was used to immunize rabbits and mice to raise polyclonal antibodies. The IgG fraction of rabbit and mouse anti-Fasciola antibodies was purified. The protein content of anti-Fasciola IgG antibody, was 8.8 and 5.2mg/ml, respectively. Sandwich ELISA was performed to detect Fasciola antigens in serum samples collected from 248 cattle. Also, a Dot blot was performed to detect Fasciola antigens in 38 human sera. Results showed that after parasitological stool examination, 26 human samples were positive and 12 human sera were healthy. The redbrown color appeared with all infected samples only. This technique is cheap, and saves time; multiple samples can make at the same time and not require expensive laboratory equipment. In conclusion, the sandwich ELISA and Dot blot assays were more reliable tools for early serodiagnosis of fasciolosis than traditional methods. Significance Statement: Different antigenic fractions of Fasciola have been used for serological diagnosis of human fasciolosis. [ABSTRACT FROM AUTHOR]

Chuquiraga weberbaueri “amaro” (Asteraceae) is a plant species popularly used for the treatment of animal fasciolosis in Peru. However, biological and chemical studies of this species remain scarce. Hence, the objective of the present study was to evaluate the anthelmintic effect of the hydroethanolic and aqueous extracts of C. weberbaueri leaves on Fasciola hepatica, through a controlled and a therapeutic efficacy test in artificially infected sheep. In addition, the chemical composition of the extracts was evaluated using phytochemical screening and analysis by UHPLC-MS/MS. Among the evaluated extracts, the aqueous extract proved to be the most efficient and of low toxicity against F. hepatica, inhibiting about 70% of its oviposition, using a 100 mg/kg dose. The chemical study showed that the major constituents were phenolic derivatives, such as chlorogenic acid and ethyl caffeate, with the notable presence of flavonoids, saponins, and lactones. Therefore, it was possible to observe the antihelmintic effect of the C. weberbaueri on the F. hepatica, which suggests the use of this plant extract as a potential alternative in the sheep fasciolosis treatment. [ABSTRACT FROM AUTHOR]

Simple Summary: Infection of humans and animals with Fasciola spp. poses a threat to both the health of humans and animals and the animal industry. In the pastoral areas of the Qinghai–Tibet Plateau, yak is the most important economic animal, especially for the herdsmen and their animal products. However, the infection of yaks with Fasciola spp. is often underestimated. In this study, we investigated a total of 1784 yaks in 16 administrative counties in the Qinghai–Tibet Plateau region by fecal examination and autopsy of yaks after slaughter, and the differences in infection rates between different age groups and regions were analyzed. The clustering characteristics of the infection rates in spatial statistics were summarized. This investigation revealed that yaks on the Qinghai–Tibet Plateau had a high Fasciola spp. prevalence (17.32%), age was a risk factor for Fasciola spp. infection, and regional differences were the other risk factor for Fasciola spp. infection. The objective of this investigation was to understand the epidemiology of fascioliasis in yaks in the alpine pastoral areas of the Qinghai–Tibet Plateau, China. The prevalence of Fasciola hepatica infection was estimated by examining eggs in the feces of yaks and by autopsy after the slaughter. Yaks were sampled from a total of 16 representative counties in Qinghai province, and risk factors were assessed based on regional and age characteristics. Fecal samples were obtained from 1542 yaks aged 0–1 (<1 year old), 1–2 (≥1 year old and <3 years old), and over 3 years (≥3 years old). In addition, 242 yaks over 3 years old who had not undergone fecal examinations were randomly selected for autopsy. A total of 267 fecal samples were positive for Fasciola spp. eggs. The average infection rate was 17.32% (0–60.61%), and the average infection intensity was 51.9 eggs per gram (epg) of feces, with intensities ranging from 18 to 112 epg. In Maduo, Dari, Zhiduo, Chengduo, and Datong counties, the Fasciola spp. eggs infection rate was zero. Fasciola spp. adult flukes were detected in 66 out of 242 yaks at autopsy, with a total infection rate of 27.27% and an average infection intensity of 21.2 (adult worms), with intensities ranging from 3 to 46 worms. Logistic regression model analysis showed that age was a significant risk factor for yak infection with Fasciola spp. In addition, the risk varied between regions: Haiyan, Gangcha, Duran, and Wulan were all high-risk areas for yak infection with Fasciola spp. The spatial distribution of the Fasciola spp. infection rate in each region showed a very weak negative correlation (Moran's I = −0.062), Duran formed a spatial distribution of high–low clusters with surrounding areas, and Datong formed a low–high clustering distribution characteristic with the surrounding areas. This investigation revealed that the infection rate of Fasciola spp. in yaks was higher on the Qinghai–Tibet Plateau. Increasing age was a risk factor for infection with Fasciola spp.; different regions also have a different risk of Fasciola spp. infection. Only two regions showed clustering characteristics in the spatial distribution of infection rates. These findings extend the epidemiological information on Fasciola spp. infection in yaks and provide baseline data for the execution of control measures against Fasciola spp. infection. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize the Tunisian Fasciola spp. flukes by morphometric and molecular analyses. Flukes were collected from livers of sheep slaughtered in Sejnane slaughterhouses (Bizerte gouvernorate, Northwest Tunisia) between January and March 2021. Five morphometric parameters were determined for all the liver flukes, as follows: (i) total body length (BL), (ii) distance between ventral sucker and the tail (VS-T), (iii) distance between oral sucker and ventral sucker (OS-VS), (iv) abdomen diameter (AD), (v) tail diameter (TD) and the body length to width ratio (BL/BW). Molecular identification of the fluke specimens was carried out by polymerase chain reaction, restriction fragment polymorphism (PCR-RFLP) of a 680 bp sequence of the internal transcribes spacer 1 (ITS1) gene and by amplification, sequencing, and phylogenetic analysis of a 500 bp sequence of the ITS2 gene. Morphometric measurements showed that the mean of the total body length of the adult flukes was 21.1 ± 2.7 mm with minimum and maximum lengths of 13 and 31 mm, respectively. The PCR-RFLP analysis revealed a single profile consisting of three bands of approximately 370, 100, and 60 bp. Fasciola sequences described in the present study (GenBank numbers: OQ457027 and OQ457028) showed 99.58–100% identity to Fasciola hepatica. In conclusion, the results of this study show that molecular and phylogenetic analyses confirm the presence of a single species of F. hepatica in the Sejnane region Northwest of Tunisia. However, further studies are needed to identify the occurrence of Fasciola species in other Tunisian regions. [ABSTRACT FROM AUTHOR]

Malacological and parasitological studies were conducted from April 2020 to March 2021 to determine the abundance and distribution of molluscs and cercariae of Schistosoma spp and Fasciola gigantica. Collected molluscs are exposed to strong light to induce cercarial release. Mollusc densities were higher at station 1 (Gamak) than in station 8 (Patakai), with Bellamya unicolor and Biomphalaria pfeifferi more abundant and Bulinus truncatus, B. tropicus, and B. globosus less abundant. The overall prevalence of cercariae (19.87%) is higher in station 3 (Yaye orchard), station 9 (Gougni), station 4 (Madiogo), station 5 (Madiogo pasture), and station 6 (Ziam 3). It varies significantly between 15.76% in station 8 and 25.77% in station 3, between 8.48% in B. truncatus and 25.53% in B. globosus, and between 19.27% for cercariae of Schistosoma spp and 21.60% for those of F. gigantica. Cercarial emissions in L. natalensis and B. pfeifferi were higher in hot and cold dry seasons; on the other hand, cercarial emissions in B. globosus were higher in hot dry seasons (31.48%) and rainy seasons (23.38%). Emissions of cercariae from S. haematobium are related to areas of human activity and defecation, while those of F. gigantica in L. natalensis, Schistosoma haematobium in B. tropicus, and S. mansoni in B. pfeifferi are related to grazing areas. Mayo-Vreck is a site that favors the endemicity of fascioliasis and human schistosomiasis. [ABSTRACT FROM AUTHOR]

Trematodes are chronic, debilitating diseases in livestock, causing significant economic losses worldwide. From mid-December 2021 to May 2022, a cross-sectional study was carried out in the Damot Sore District to estimate the prevalence of trematode infections in cattle and associated risk factors. Trematode eggs were found in 100 of the 384 faecal samples tested, with an overall prevalence of 26.04% (95% CI: 21.88-30.69%). The prevalence for Fasciola, Paramphistome, and Schistosoma species were 12.50%, 5.21%, and 0, respectively. Moreover, the infection rate with two parasites was 8.33%. The data were then examined further using univariable and multivariable logistic regression analysis. As a result, age was the only potential predictor identified to influence trematode infections in cattle among the potential predictors considered. Furthermore, old cattle were more likely to be infected with trematodiasis nearly 12 times (OR = 11.5) that of young cattle, and this difference was statistically significant (p < 0.05), whereas other risk factors considered were statistically insignificant (p > 0.05). According to the findings of this study, cattle trematodiasis is a moderately common disease in the study area. As a result, additional research on the meteorological conditions of snail infection was forwarded, along with other points to reduce the disease problem in livestock production. [ABSTRACT FROM AUTHOR]

Background and Aim: Parasitic diseases, including trematode invasions, result in losses to livestock in Indonesia, hindering the government's efforts to achieve meat self-sufficiency. This study aimed to estimate the prevalence of Amphistomes and Fasciola in large ruminants reared by smallholder farmers. Materials and Methods: Fecal samples from 199 buffalo and cattle were collected from the districts of East Lampung (Lampung Province) and Lebak (Banten Province). Fecal samples were examined for the presence of trematode eggs using a sedimentation technique. Results: Parasite invasion rate was 48.2% (95% confidence interval [CI]: 41.3%-55.2%). Rate of invasion was 63.3% (95% CI: 52.7%-73.9%) in Lampung and 38.3% (95% CI: 29.6%-47.0%) in Lebak-Banten. The prevalence of multiple invasions of both Amphistomes and Fasciola was 20% in buffalo and local cattle, whereas invasion rate was 12.8% in crossbred cattle. Invasion rate of Amphistomes alone was 27.1%, and that of Fasciola was 4.5%. A higher invasion rate of Amphistomes (29.8%) occurred in crossbred animals. There were no significant differences between age groups for trematode invasion. The Chi-square test showed that the prevalence of trematode invasion in females was significantly higher than in males (51.5% and 30.0%, respectively). Amphistomes more commonly infected females than males (29.0% and 16.7%, respectively). Conclusion: All breeds were vulnerable to invasion by both trematode species and single invasions with different invasion rates. These findings contribute to determining the magnitude of the disease and provide a basis for studies on prevention and treatment of trematode invasion. [ABSTRACT FROM AUTHOR]

Background: Fasciolosis is a foodborne disease caused by Fasciola sp. infecting ruminants, especially cattle. Fasciolosis remains a significant concern for Veterinary Public Health because of its zoonosis risk and transmission mode. Aim: This study aimed to determine the prevalence and risk factors associated with Fasciola gigantica infestation in cattle at Ampel abbatoir, Central Java, Indonesia. Methods: A cross-sectional study was performed on 585 cattle from February to August 2022. Visual observation postmortem was used to assess Fasciola infection based on adult flukes in liver parenchyma and ductuli biliferi. Results: The overall prevalence of fasciolosis in Ampel abbatoir is high, reaching 25.12% (147/585). The highest prevalence was observed in the Ongole breed, 42.1% (24/57), female cattle, 38.72% (115/297), body condition score criteria of 2 50% (21/42), cattle aged >3.5 years 46.06% (82/178), and cattle originated from outside of Boyolali district 33.33% (71/213). Conclusion: This study showed a high prevalence of fasciolosis in Ampel abbatoir, as shown in the correlation between the risk factors of breed, sex, body condition score (BCS), origin, and age. Because of the high prevalence of fasciolosis in the abattoirs, it is essential to continue performing epidemiology studies in more expansive areas. The subsequent plans are important to reduce the risk of fasciolosis as a threat to productive cattle husbandry and warrant its transmission to humans as a foodborne-zoonotic disease. [ABSTRACT FROM AUTHOR]

Background: Fasciolosis is a parasitic disease affecting the hepatobiliary system of livestock worldwide. The control of the fluke is important to be performed in endemic regions. Aim: This study aims to evaluate the effect of Etlingera elatior ethanolic extract on egg and adult stadia of Fasciola gigantica. Methods: Fasciola gigantica in different stages were incubated with E. elatior ethanolic extract in different concentrations and time points. Results: The number of developed eggs with different concentrations of 1.25%, 2.5%, and 5% was significantly decreased by 36.67%, 56.67%, and 56.67% on day 11 post-incubation, which showed an ovicidal effect of the herb. The developed eggs on day 14, which were represented by hatched larvae, were also decreased by 70%, 50%, and 13.33%, respectively. Significant flukicidal effects were observed in the incubation time of 80 minutes for the concentration of 20% (p = 0.007) and 640 minutes for 10% concentration (p = 0.003). Surface microscopy of adult F. gigantica showed damaged skin and spina with the erosion of the inner membrane and detached syncytium from the tegument. Conclusion: Overall, the results indicate that E. elatior has a promising anthelmintic property against F. gigantica in both ova and adult stages. [ABSTRACT FROM AUTHOR]

The Andean bear (Tremarctos ornatus) is the only species of the Ursidae that inhabits the Andes in South America. The natural history of the species and health issues related to infectious diseases, particularly the presence of gastrointestinal parasites (GIP), is unknown. To determine the prevalence, infestation, and richness of GIP in fecal samples of wild Andean bears, 21 fecal samples were collected at different sites of the Chingaza National Park, Colombia, and its buffer area from February to April of 2019. Samples were analyzed by qualitative and quantitative coprological techniques. A minimum of one parasitic species was present in 100% (n = 21) of the samples and in 86% of them, co-infection was detected. In decreasing order, the observed parasite prevalence was as follows: 100% for Cryptosporidium spp.; 62% for Ascaris spp.; 38% for Baylisascaris spp.; 33% for Trichomonas spp.; 19% for Strongylus spp.; 14% for Microsporidia and Coccidian oocysts; followed by Trichostrongylus spp. and ciliated protozoans with 10%; and Ancylostoma spp., Blastocystis spp., and Fasciola spp. were present in 5% of the samples. The presence of Blastocystis spp., Baylisascaris spp., Ascaris spp., and Cryptosporidium spp. suggests a possible rotation of parasitic species between domestic and wild host species. This is the first report of GIP for Andean bears in the east range of Colombia, which could help to generate other studies in the country and South America. [ABSTRACT FROM AUTHOR]

Background: Fascioliasis, caused by the liver flukes Fasciola hepatica and Fasciola gigantica, is a global zoonotic helminthic disease. The livestock and human are the final hosts of the parasites. Northern Iran is an important endemic region for fascioliasis. Few studies have been conducted on the characterization of Fasciola isolates from eastern regions of the Caspian littoral of the country. Objective: The aim of the present study was to identify F. hepatica, F. gigantica and intermediate/hybrid forms of Fasciola isolates from livestock in Golestan province, northern Iran, using morphometric and molecular tools. Methods: Livestock livers naturally infected with Fasciola spp. were collected from Golestan slaughterhouse during 2019–2020. The worms were morphometrically studied using a calibrated stereomicroscope. Genomic DNA was extracted from all samples, and polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was performed on internal transcribed spacer (ITS1) region using Rsa1 restriction enzyme. All the isolates were then analysed by multiplex PCR on Pepck region. Results: A total of 110 Fasciola isolates were collected from the infected livers, including 94 sheep, 12 cattle and 4 goats. Morphometric analysis of 61 adult Fasciola isolates indicated that, 44 and 17 isolates belonged to F. hepatica and F. gigantica, respectively. Eighty‐one and 29 isolates belonged to F. hepatica and F. gigantica using ITS1‐RFLP, respectively. However, Pepck Multiplex PCR indicated 72 F. hepatica, 26 F. gigantica and 12 intermediate/hybrid forms. All 12 hybrid isolates were found in sheep host. Two isolates were identified as F. gigantica using morphometry and F. hepatica using both molecular methods. Conclusion: The present study confirmed the existence of both F. hepatica and F. gigantica species and reported the first molecular evidence of hybrid Fasciola isolates in ruminants of Golestan province. [ABSTRACT FROM AUTHOR]

Background and Aim: Fasciolosis due to Fasciola gigantica is endemic to tropical countries and Fasciola hepatica in temperate climates, highly detrimental to livestock and known as foodborne zoonotic diseases. The strategic control of the disease is mainly the use of chemical anthelmintic. This study aimed to evaluate the anthelmintic properties of Artemisia vulgaris extract on the ova and adult stages of F. gigantica. Materials and Methods: Samples were collected from the Ampel Abbatoir, Boyolali District, Central Java, Indonesia. The ova from 20-gallbladders of cattle which were naturally infected with F. gigantica and 270 living F. gigantica worms were used in this study. The ovicidal assay was performed by incubating the ova with A. vulgaris in different concentrations, that is, 5%, 2.5%, and 1.25% for 5, 9, 11, 14, and 16 days. The efficacies were evaluated by quantification of ova degeneration during developmental stages in different time points and egg-hatch assay. The flukicidal effects were observed by mortality assay in 5, 10, 20, 40, 80, 160, 320, and 640 min incubations followed by scanning electron microscopy for surface morphology and histology of the fluke's transversal sections. Results: The concentration of 5% A. vulgaris showed the strongest ovicidal activities. The percentage of hatching ova on day 16 at concentrations of 5%, 2.5%, and 1.25% were 3.33%, 6.67%, and 16.67%. These ova hatch assay showed a significant reduction (p < 0.001) compared to untreated control. The flukicidal effect was significant (p < 0.001) at a concentration of 20%, with a mortality rate reaching 66.67% in the 40 min of incubation time. The surface properties of the adult worms, including the spine, tegument, acetabulum, intestine, and vitelline follicles, were disintegrated. Conclusion: The results showed that A. vulgaris has the potential ovicidal and flukicidal properties to F. gigantica. The active compounds remained necessary to be elucidated further and its modes of action would be interesting to be predicted by molecular docking modeling. [ABSTRACT FROM AUTHOR]

Fasciola gigantica, responsible for the zoonotic disease fasciolosis, pose a great threat to the livestock and human health worldwide. The triclabendazole (TCBZ) has been used for decades as a broad spectrum anthelmintic to control this perilous disease but the emergence of resistance in flukes against TCBZ has prompted researchers across the world to explore for new drugs and antigenic targets. World Health Organization has strongly recommended the utilization of neurobiologically significant biomolecules as new drug/antigenic targets because of their significant role in the physiology of parasites. Monoamine Oxidase (MAO) is an important neurobiological enzyme which catabolizes aminergic neurotransmitters thus preventing prolonged excitation of neurons and in non-neuronal cells it prevents cellular toxicity due to accumulation of toxic monoamines. Owing to the important role of MAO in the survival and perpetuation of parasites, multipronged approaches were undertaken for the characterization of MAO-A in F. gigantica. The activity of MAO was found to be 1.5 times higher in the mitochondrial samples than the whole homogenate samples. The adult worms of the F. gigantica appeared to possess both the isoforms of MAO i.e., MAO-A and MAO-B. The zymographic studies revealed strong enzyme activity in its native state as assessed through prominent dark bands at 250KDa in the zymogram. The enzyme was also found to be highly immunogenic as revealed by high antibody titer at 1:6400 dilution. The immunogenicity of MAO-A enzyme was further established in the Western Blots in which a strong band of 50KDa was distinctly evident. Despite ubiquitous presence of MAO in F. gigantica some regions like tegumental surface and intestinal caecae displayed strong immunofluorescence as compared to other regions. The detection of MAO-A in the F. gigantica samples in Dot-Blot assay indicate a great potential of this molecule for the immunodiagnostics of fasciolosis, particularly in the field conditions. The enzyme activity was sensitive to the specific inhibitor clorgyline in a concentration dependant manner, particularly in the late incubation period. The zymographic results also exhibited similar trend. The strong intensity of spots in Dot-blots indicate high immunogenicity of the MAO protein. The intensity of bands/spots in the samples of worms treated with clorgyline also declined, clearly indicating that the tropical liver fluke possesses prominent MAO-A activity. [ABSTRACT FROM AUTHOR]

Introduction: Fasciolosis represents a significant economic impact on livestock production in different countries. The gold standard method for diagnosing fasciolosis is currently a coprological examination, based on egg detection in stool. However, this approach is undoubtedly ineffective, especially during the acute phase of the disease. In order to detect all stages of infection, an alternative serological method is used to confirm the infected animals with fasciolosis. Thus, we aimed to identify the pre-patent and patent periods of equine fasciolosis using both copro-microscopical and the ELISA technique in different areas in Garmian region and Sulaimani provinces. Material and Methods: A coprological and the sensitive ELISA techniques were used to detect the presence of Fasciola hepatica in the infected horses through detection of the serum antibodies against secretary and the excretory antigens from F. hepatica. Results: The copro-prevalence and seroprevalence of equine fasciolosis were found to be 16% and 58%, respectively, in a total of 50 fecal and 50 blood samples from equine of various ages. Conclusion: It was concluded that a serological test had an accurate result for the detection of Fasciola infection in horses than the coprological methods; particularly in horses had advanced into the patent period of the infection. [ABSTRACT FROM AUTHOR]

This study was conducted for the purpose of evaluating infection with the parasite Fasciola spp in the city of Mosul for the period from June 2020 to June 2022. The results showed that a total of 12.30% of cattle and sheep had Fasciola eggs in their feces, with a significant difference between cattle and sheep, and the infection was high in cattle 16%. The eggs of the Fasciola species are oval, brown to yellow in color, and have an operculum. There was no significant difference in the rate of infection with the Fasciola parasite and the sex of cattle and sheep. Regarding the relationship between the rate of infection and the age of the animals infected, there were no significant differences between the infection and the age groups of the examined sheep. Whereas there was a significant difference between the cattle whose age was less than one year, one to two years old and more than two years old. The results of the serological examination of 90 blood samples of cattle and sheep showed that the seroprevalence of Fasciola in both cattle and sheep was 22.22% and 8.88%, respectively, with no difference in the incidence of infection between all the examined cattle and sheep. [ABSTRACT FROM AUTHOR]

M., which received a (C), was the study's main objective. The whole thing was eaten. a couple of quotes from (C. dispersed in a liquid. has antibacterial properties (E. elegans). It's necessary to remove the body. C. Eczema on the face is comparable to this. Sheep and novyi both had organic infections. novyi. Following different combinations of treatments, a heavy dose of treatment, and a light dose, fasciolitis manifested in all three groups of infected sheep. The treatment groups received an oral 150 mg/kg aqueous M. oleifera leaf extract every 48 hours for 21 days. Body mass, fecal egg counts, blood anti-Fasciola IgG levels, cytokines (IL-2, IL-17, and IL-10), and C. the elegans' bacterial inhabitants jokingly strike up a conversation with everyone there. Sheep with light and heavy infections gained more weight after receiving M. oleifera treatment and had lower fecal egg counts than control groups; sheep with light infection had a 100% reduction in egg count. The serum concentrations of IL-2, IL-17, and IgG all significantly decreased after M. oleifera treatment. There were significantly higher levels of IL-10 in sheep with both mild and severe infections. C's feces contained noticeably fewer bacteria after taking moringa extract. The two novyi groups had varying degrees of infection severity. Fabricio (F. (as well as OleiferaMdot). C. noteworthy and novyi. M. oleifera aqueous extracts showed a concentration-dependent ovicidal effect on F. gigantica non-embryonated. [ABSTRACT FROM AUTHOR]

Background: Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica(F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution for the genetic diversity of the Fasciola parasite whose genomes are ∼1.3 GB in size. Objectives: To gain a better understanding of the gene diversity of Fasciola isolates from western Iran and to identify the most informative markers as candidates for epidemiological studies, five housekeeping genes were evaluated using a multilocus sequence typing (MLST) approach. Methods: MLST analysis was developed based on five genes (ND1, Pepck, Pold, Cyt b and HSP70) after genomic DNA extraction, amplification and sequencing. Nucleotide diversity and phylogeny analysis were conducted on both concatenated MLST loci and each individual locus. A median joining haplotype network was created to examine the haplotypes relationship among Fasciola isolates. Results: Thirty‐three Fasciola isolates (19 F. hepatica and 14 F. gigantica) were included in the study. A total of 2971 bp was analysed for each isolate and 31 sequence types (STs) were identified among the 33 isolates (19 for F. hepatica and 14 for F. gigantica isolates). The STs produced 44 and 42 polymorphic sites and 17 and 14 haplotypes for F. hepatica and F. gigantica, respectively. Haplotype diversity was 0.982 ± 0.026 and 1.000 ± 0.027 and nucleotide diversity was 0.00200 and 0.00353 ± 0.00088 for F. hepatica and F. gigantica, respectively. There was a high degree of genetic diversity with a Simpson's index of diversity of 0.98 and 1 for F. hepatica and F. gigantica, respectively. While HSP70 and Pold haplotypes from Fasciola species were separated by one to three mutational steps, the haplotype networks of ND1 and Cyt b were more complex and numerous mutational steps were found, likely due to recombination. Conclusions: Although HSP70 and Pold genes from F. gigantica were invariant over the entire region of sequence coverage, MLST was useful for investigating the phylogenetic relationship of Fasciola species. The present study also provided insight into markers more suitable for phylogenetic studies and the genetic structure of Fasciola parasites. [ABSTRACT FROM AUTHOR]

Simple Summary: The 22nd chromatography component (F22) of the Fasciola gigantica excretory-secretory products (FgESP) shows better diagnostic value than the FgESP, but its immunomodulatory function and potential as a molecular vaccine candidate are unknown. Thus, the effect of F22 on the mitogen-induced proliferation of buffalo peripheral blood mononuclear cells (PBMCs) in the innate immune response was preliminarily studied, and its components were also explored. The results indicated that F22 significantly decreased the proliferation of PBMCs stimulated with mitogen. Two hundred and sixteen were identified as components of the F22, and these included eighty-six proteins present in more than one pathway and some with robust immunomodulatory ability. Further studies should be performed to investigate the immunomodulatory function of F22 in the adaptive immune response, and the components of F22 can be further studied as potential vaccine candidate molecules. The 22nd chromatography component (F22) of the Fasciola gigantica excretory-secretory products (FgESP) shows better diagnostic value than the FgESP, and diagnostic methods based on F22 have also been established. Thus, exploring its immunomodulatory function and potential as a molecular vaccine candidate is attractive. In the present study, the effect of F22 on the mitogen-induced proliferation of buffalo peripheral blood mononuclear cells (PBMCs) in the innate immune response was preliminarily studied using the FgESP as a control. PBMCs were incubated with concanavalin A (ConA) and phytohemagglutinin (PHA) at optimal (1 µg/well) or suboptimal (0.25 µg/well) doses coupled with FgESP and F22 at different doses (1–16 µg/well). Cell proliferation was then assessed by microenzyme reaction colorimetry (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) assay). In addition, the components of F22 were also explored by mass spectrometry and then subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to infer their functions. The results indicated that FgESP decreased the proliferation of PBMCs stimulated with ConA and PHA at specific doses, whereas F22 significantly decreased the proliferation of PBMCs stimulated with ConA and PHA at both optimal and suboptimal doses (p < 0.05). Two hundred and sixteen proteins were identified in F22, and these included 86 proteins that could be assigned to more than one pathway and some with robust immunomodulatory ability. Further studies should be performed to investigate the immunomodulatory function of F22 in the adaptive immune response, and the components of F22 can be further studied as potential vaccine candidate molecules. [ABSTRACT FROM AUTHOR]

Background: Extracellular vesicles (EVs) released by helminths play an important role in parasite-host communication. However, little is known about the characteristics and contents of the EVs of Fasciola gigantica, a parasitic flatworm that causes tropical fascioliasis. A better understanding of EVs released by F. gigantica will help elucidate the mechanism of F. gigantica-host interaction and facilitate the search for new vaccine candidates for the control and treatment of fascioliasis. Methods: Two different populations of EVs (15k EVs and 100k EVs) were purified from adult F. gigantica culture media by ultracentrifugation. The morphology and size of the purified EVs were determined by transmission electron microscopy (TEM) and by the Zetasizer Nano ZSP high performance particle characterization system. With the aim of identifying diagnostic markers or potential vaccine candidates, proteins within the isolated 100k EVs were analyzed using mass spectrometry-based proteomics (LC–MS/MS). Mice were then vaccinated with excretory/secretory products (ESPs; depleted of EVs), 15k EVs, 100k EVs and recombinant F. gigantica heat shock protein 70 (rFg-HSP70) combined with alum adjuvant followed by challenge infection with F. gigantica metacercariae. Fluke recovery and antibody levels were used as measures of vaccine protection. Results: TEM analysis and nanoparticle tracking analysis indicated the successful isolation of two subpopulations of EVs (15k EVs and 100k EVs) from adult F. gigantica culture supernatants using differential centrifugation. A total of 755 proteins were identified in the 100k EVs. Exosome biogenesis or vesicle trafficking proteins, ESCRT (endosomal sorting complex required for transport) pathway proteins and exosome markers, heat shock proteins and 14-3-3 proteins were identified in the 100k EVs. These results indicate that the isolated 100k EVs were exosome-like vesicles. The functions of the identified proteins may be associated with immune regulation, immune evasion and virulence. Mice immunized with F. gigantica ESPs, 15k EVs, 100k EVs and rFg-HSP70 exhibited a reduction in fluke burden of 67.90%, 60.38%, 37.73% and 56.6%, respectively, compared with the adjuvant control group. The vaccination of mice with F. gigantica 100k EVs, 15k EVs, ESP and rFg-HSP70 induced significant production of specific immunoglobulins in sera, namely IgG, IgG1 and IgG2a. Conclusion: The results of this study suggest that proteins within the exosome-like vesicles of F. gigantica have immunomodulatory, immune evasion and virulence functions. This knowledge may lead to new strategies for immunotherapy, vaccination and the diagnosis of fascioliasis. [ABSTRACT FROM AUTHOR]

Background: Extracellular vesicles (EVs) released by helminths play an important role in parasite-host communication. However, little is known about the characteristics and contents of the EVs of Fasciola gigantica, a parasitic flatworm that causes tropical fascioliasis. A better understanding of EVs released by F. gigantica will help elucidate the mechanism of F. gigantica-host interaction and facilitate the search for new vaccine candidates for the control and treatment of fascioliasis. Methods: Two different populations of EVs (15k EVs and 100k EVs) were purified from adult F. gigantica culture media by ultracentrifugation. The morphology and size of the purified EVs were determined by transmission electron microscopy (TEM) and by the Zetasizer Nano ZSP high performance particle characterization system. With the aim of identifying diagnostic markers or potential vaccine candidates, proteins within the isolated 100k EVs were analyzed using mass spectrometry-based proteomics (LC–MS/MS). Mice were then vaccinated with excretory/secretory products (ESPs; depleted of EVs), 15k EVs, 100k EVs and recombinant F. gigantica heat shock protein 70 (rFg-HSP70) combined with alum adjuvant followed by challenge infection with F. gigantica metacercariae. Fluke recovery and antibody levels were used as measures of vaccine protection. Results: TEM analysis and nanoparticle tracking analysis indicated the successful isolation of two subpopulations of EVs (15k EVs and 100k EVs) from adult F. gigantica culture supernatants using differential centrifugation. A total of 755 proteins were identified in the 100k EVs. Exosome biogenesis or vesicle trafficking proteins, ESCRT (endosomal sorting complex required for transport) pathway proteins and exosome markers, heat shock proteins and 14-3-3 proteins were identified in the 100k EVs. These results indicate that the isolated 100k EVs were exosome-like vesicles. The functions of the identified proteins may be associated with immune regulation, immune evasion and virulence. Mice immunized with F. gigantica ESPs, 15k EVs, 100k EVs and rFg-HSP70 exhibited a reduction in fluke burden of 67.90%, 60.38%, 37.73% and 56.6%, respectively, compared with the adjuvant control group. The vaccination of mice with F. gigantica 100k EVs, 15k EVs, ESP and rFg-HSP70 induced significant production of specific immunoglobulins in sera, namely IgG, IgG1 and IgG2a. Conclusion: The results of this study suggest that proteins within the exosome-like vesicles of F. gigantica have immunomodulatory, immune evasion and virulence functions. This knowledge may lead to new strategies for immunotherapy, vaccination and the diagnosis of fascioliasis. [ABSTRACT FROM AUTHOR]

Continuous attempts have been made to pinpoint candidate vaccine molecules and evaluate their effectiveness in order to commercialise such vaccines for the treatment of tropical fascioliasis in livestock. The pathophysiology of fascioliasis can be related to liver damage brought on by immature flukes that migrate and feed, as well as immunological reactions to chemicals produced by the parasites and alarm signals brought on by tissue damage. Future research should, in our opinion, concentrate on the biology of invasive parasites and the resulting immune responses, particularly in the early stages of infection. The goal of the current study was to use the calcium-binding proteins from F. gigantica to create a multi-epitope subunit vaccine. The adjuvant, B-cell epitopes, CTL epitopes, and HTL epitopes that make up the vaccine construct are all connected by certain linkers. The antigenicity, allergenicity, and physiochemical properties of the vaccine construct were examined. The vaccine construct was docked with toll-like receptor 2, and simulations of the molecular dynamics of the complex's stability, interaction, and dynamics were run. After performing in silico cloning and immunosimulation, it was discovered that the construct was suitable for further investigation. New vaccination technologies and adjuvant development are advancing our food safety procedures since vaccines are seen as safe and are accepted by the user community. This research is also applicable to the F. hepatica system. [ABSTRACT FROM AUTHOR]

This study aims to investigate the common and important parasitic infections that reduce the nutritional value and also cause serious damage and disrupt the functioning of systems such as the liver and bile ducts as a vital organ of the human body and livestock. In order to investigate the mentioned parasitic infections in the autumn season, 60,881 livestock, including 32,213 sheep, 21,368 goats, 6,186 cows, 1,059 ostriches and 55 camels were slaughtered in Fasaran Slaughterhouse in 2022. Method: According to the working timetable and the existence of two slaughter lines, organs such as liver, lung, heart and carcass are examined macroscopically, then samples suspected of contamination are separated for microscopic examination. Results: The prevalence rate of hydatid cyst parasite was 49.44%, including 30.02% in sheep, 6.78% in goats, and 12.63% in cows. Moreover, the severity of infection in the liver of sheep, goats, and cows was 10, 1.04, and 2.89%, respectively. Also, the severity of lung contamination in sheep, goats, and cows was 20.02, 5.73 and 9.46% respectively. The prevalence rate of cysticercosis parasite was 0.38%, in a way that, infection rate in the carcasses of sheep, goats, and cows was 0.27%, 0.60%, and 0.40%, respectively. Conclusion: According to the results, it can be concluded that parasitic infection can play an important role in reducing the nutrients of infected tissues, and in case of human infection, pathological complications can affect the functioning of body organs and disrupt and cause serious harm to human health. [ABSTRACT FROM AUTHOR]

Background: Fascioliasis is a significant vector-borne disease that has emerged in numerous tropical and subtropical countries causing severe health problems. Egypt is one of the fascioliasis endemic regions; however, the current situation in Upper Egypt is understudied, with only sporadic human cases or outbreaks. This study aims to highlight the sociodemographic characteristics of human fascioliasis in a newly emerged endemic area in Upper Egypt, along with risk factors analysis and the molecular characteristics of the fasciolid population in humans, animals, and lymnaeid snails. Methodology/Principal findings: The study reported Fasciola infection in patients and their close relatives by analyzing the risk of human infection. Morphological and molecular characterization was performed on lymnaeid snails. Multigene sequencing was also used to characterize fasciolids from human cases, cattle, and pooled snail samples. The study identified asymptomatic Fasciola infection among family members and identified the presence of peridomestic animals as a significant risk factor for infection. This is the first genetic evidence that Radix auricularia exists as the snail intermediate host in Egypt. Conclusions/Significance: This study revealed that Assiut Governorate in Upper Egypt is a high-risk area for human fascioliasis that requires additional control measures. Fasciola hepatica was the main causative agent infecting humans and snail vectors in this newly emerged endemic area. In addition, this is the first report of R. auricularia as the snail intermediate host transmitting fascioliasis in Upper Egypt. Further research is required to clarify the widespread distribution of Fasciola in Egypt's various animal hosts. This provides insight into the mode of transmission, epidemiological criteria, and genetic diversity of fasciolid populations in Upper Egypt.\ Author summary: Human fascioliasis is a freshwater snail-transmitted disease that is widely distributed in many tropical and subtropical countries including Egypt. Recently, Upper Egypt reported sporadic cases of human fascioliasis and a few outbreaks establishing a new endemic area. This study was conducted to get information on the sociodemographic characteristics of the infected patients and possible risk factors of infection in this recently emerged endemic region. Further, this study tried to describe lymnaeid intermediate host snails involved in infection transmission morphologically and by molecular analysis. Analysis of the phylogenetic relationships between natural fasciolid populations was performed in humans, animals, and snail isolates by multilocus sequencing. Results indicated that asymptomatic fascioliasis should be screened in areas at risk of infection. Also, younger age groups and the presence of peri domestic animals were significantly associated with an increased risk of infection. This is the first molecular proof of the existence of Radix auricularia snails causing fascioliasis in Egypt. Results showed that Upper Egypt may be an area at risk for human fascioliasis that needs additional control measures. [ABSTRACT FROM AUTHOR]

In the past decade significant advances in our understanding of liver fluke biology have been made through in-depth interrogation and analysis of evolving Fasciola hepatica and Fasciola gigantica omics datasets. This information is crucial for developing novel control strategies, particularly vaccines necessitated by the global spread of anthelmintic resistance. Distilling them down to a manageable number of testable vaccines requires combined rational, empirical, and collaborative approaches. Despite a lack of clear outstanding vaccine candidate(s), we must continue to identify salient parasite–host interacting molecules, likely in the secretory products, tegument, or extracellular vesicles, and perform robust trials especially in livestock, using present and emerging vaccinology technologies to discover that elusive liver fluke vaccine. Omics tools are bringing this prospect ever closer. The generation of extensive omics datasets for Fasciola hepatica and Fasciola gigantica has expedited the investigation of key molecules involved in liver fluke biology, pathogenicity, and virulence that can be targeted for vaccine development. Vaccines are urgent to counteract the spread of anthelminthic resistance; however, very few molecules have been brought forward into large animal vaccine trials, particularly on the field. Discovery of effective vaccine antigen cocktails from the large number of genes/proteins within publicly available omics datasets will require a strategy combining both rational, empirical, and collaborative approaches. A multiantigen approach, based on molecules important for key processes in the early-stage parasites, may form a key strategy for the development of a fasciolosis vaccine, aimed at preventing the extensive damage caused by the migrating parasite. [ABSTRACT FROM AUTHOR]

Liver flukes, Fasciola spp., are veterinary and medically important parasites infecting numerous species of economically important animals in addition to humans on a global scale. The components of transforming growth factor beta (TGF-β) signalling are widely distributed throughout the animal kingdom and are considerably conserved. Through shared common signal transduction mechanisms, crosstalk of TGF-β signalling between a host and the parasite during infection is possible. Herein, we have identified and undertaken the molecular characterisation of a putative TGF-β homologue from the tropical liver fluke F. gigantica (FgTLM). A FgTLM cDNA was 3557 bp in length, it encoded for 620 amino acid polypeptide which consisted of 494 amino acids of prodomain and 126 amino acids comprising the mature protein. FgTLM displayed characteristic structures of mammalian TGF-β ligands that were unique to the inhibin-β chain, monomer of activin. A phylogenetic analysis revealed the high degree of conservation with TGF-β molecules from trematode species. Interestingly, the sequence of amino acid in the active domain of FgTLM was completely identical to FhTLM from F. hepatica. FgTLM expressed throughout the lifecycle of F. gigantica but was highly expressed in developmental active stages. The dynamics of expression of FgTLM during the developmental stages of F. gigantica was comparable to the pattern of TGF-β expression in F. hepatica. Our findings demonstrated that FgTLM exhibits a high level of similarity to FhTLM in the context of both amino acid sequence and the life stage expression patterns. These similarities underline the possibility that the FgTLM molecule might have the same properties and functions as FhTLM in biological processes of the immature parasites and host immune evasion. Consequently, the specific biological functions of FgTLM on either parasite or relevant hosts need to be defined experimentally. [ABSTRACT FROM AUTHOR]

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 first chimeric protein (rFgCHI) with three different proteins was constructed. • Anti-rFgCHI antibody recognized the liver fluke at all stages. • Anti-rFgCHI antibody could detect the native tissue on cecal epithelium. • Cross-reactivity of anti-rFgCHI were not found in the other related parasites. • This study highlighted the key design criteria for chimeric protein production. [ABSTRACT FROM AUTHOR]

Background and Aim: Indonesia’s farming practices are a perfect setting for establishing an infection with Fasciola gigantica which can result in economically detrimental. The objectives of the current study were to describe and analyze the transmission dynamics of fasciolosis (F. gigantica) in Prafi District, to provide information on effective control strategies and to identify risk factors associated with fasciolosis in cattle. Materials and Methods: Fecal samples were purposively collected from the rectum of 60 beef cattle in Prafi District, Manokwari Regency, West Papua Province, Indonesia. The samples were collected once a month for 8 months from April 2019 to November 2019. Furthermore, the samples were taken from two rearing system groups: 30 stall cattle and 30 cattle in a palm oil garden. The presence of F. gigantica eggs in the fecal samples was examined using a modified Danish Bilharziasis Laboratory technique-sedimentation. Meanwhile, the antigenic diagnosis of Fasciola in the fecal samples was analyzed using an enzyme-linked immunosorbent assay kit to perform an indirect sandwich assay on feces. Snails were collected from an irrigation canal, rice field, and palm oil garden canal around Prafi District. These snails were examined for infection with cercariae larvae of F. gigantica by cercarial shedding and crushing techniques. Results: The peak occurrence of F. gigantica infection was identified in August (65.00%) and the lowest in June (35.00%). The highest prevalence of fasciolosis in cattle was recorded in August and November (90.00%) and the lowest was in May (40.00%). Moreover, the highest prevalence of fasciolosis in cattle exposed to the palm oil garden was recorded in April (53.33%) and the lowest prevalence of F. gigantica infection was recorded in June (23.33%). In total, 2046 snails were screened by the cercarial shedding and crushing method; of these, 426 (20.82%) were found to be positive for trematode cercariae. The prevalence of F. gigantica infection in Lymnaea spp. snails was highest in November (47.46%) and lowest in April (9.28%). Conclusion: The current study shows that beef cattle grown in two types of rearing systems in Prafi District were infected with F. gigantica during the dry and rainy season. It was revealed that Lymnaea spp. are common snails found in and around Prafi District, and can act as intermediate hosts with an infective stage of trematode. The transmission to cattle was highly effective, despite the short activity period, the low infection rate of snails, and the incidental anthelminthic treatment. [ABSTRACT FROM AUTHOR]

Background: Multiplex polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) for nuclear phosphoenolpyruvate carboxykinase (pepck) and polymerase delta (pold), respectively, have been used to differentiate Fasciola hepatica, F. gigantica, and hybrid Fasciola flukes. However, discrimination errors have been reported in both methods. This study aimed to develop a multiplex PCR based on a novel nuclear marker, the fatty acid binding protein type I (FABP) type I gene. Methods: Nucleotide sequence variations of FABP type I were analyzed using DNA samples of F. hepatica, F. gigantica, and hybrid Fasciola flukes obtained from 11 countries in Europe, Latin America, Africa, and Asia. A common forward primer for F. hepatica and F. gigantica and two specific reverse primers for F. hepatica and F. gigantica were designed for multiplex PCR. Results: Specific fragments of F. hepatica (290 bp) and F. gigantica (190 bp) were successfully amplified using multiplex PCR. However, the hybrid flukes contained fragments of both species. The multiplex PCR for FABP type I could precisely discriminate the 1312 Fasciola samples used in this study. Notably, no discrimination errors were observed with this novel method. Conclusions: Multiplex PCR for FABP type I can be used as a species discrimination marker in place of pepck and pold. The robustness of the species-specific primer should be continuously examined using a larger number of Fasciola flukes worldwide in the future since nucleotide substitutions in the primer regions may cause amplification errors. [ABSTRACT FROM AUTHOR]

Fascioliasis is a zoonotic disease caused by Fasciola hepatica or F. gigantica. There are two phases of Fasciola infection. In the acute phase, the patient has a wide spectrum of clinical manifestations while the later phase of infection is usually related to inflammatory changes in the bile duct and the mechanical effects of the worm, which can lead to biliary obstruction. The presence of Fasciola inside the biliary system has rarely been reported in the literature. In this report, we presented a case of bile duct obstruction in a 36-year-old male patient undergoing cholecystectomy for gallstones and with a history of worsening jaundice for one month. Extensive laboratory testing failed to clarify the cause of jaundice. Three living liver flukes were subsequently retrieved from the right hepatic duct and later confirmed as F. gigantica by morphology and internal transcribed spacer (ITS2) sequence analysis. [ABSTRACT FROM AUTHOR]