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Fasciola hepatica causes liver fluke disease, a worldwide neglected and re-emerging zoonotic disease, leading to hepatitis in humans and livestock. In the pathogenesis, flukes actively migrate through liver parenchyma provoking tissue damage. Here, parasites must confront leukocytes of the innate immune system in vivo. Polymorphonuclear neutrophils (PMN) are the most abundant granulocytes and first ones arriving at infection sites. PMN may display neutrophil extracellular traps (NETs), consisting of nuclear DNA, decorated with histones, enzymes, and antimicrobial peptides. We investigated for the first time whether F. hepatica soluble antigens (FhAg) can also trigger NETosis and innate immune reactions in exposed ovine PMN. Thus, isolated PMN were co-cultured with FhAg and NET formation was visualized by immunofluorescence and scanning electron microscopy analyses resulting in various phenotypes with spread NETs being the most detected in vitro. In line, NETs quantification via Picogreen®-fluorometric measurements revealed induction of anchored- and cell free NETs phenotypes. Live cell 3D-holotomographic microscopy revealed degranulation of stimulated PMN at 30 min exposure to FhAg. Functional PMN chemotaxis assays showed a significant increase of PMN migration (p = 0.010) and intracellular ROS production significantly increased throughout time (p = 0.028). Contrary, metabolic activities profiles of FhAg-exposed PMN did not significantly increase. Finally, in vivo histopathological analysis on F. hepatica-parasitized liver tissue sections of sheep showed multifocal infiltration of inflammatory cells within liver parenchyma, and further fluorescence microscopy analyses confirmed NETs formation in vivo. Overall, we hypothesized that NET-formation is a relevant host defence mechanism that might have a role in the pathogenesis of fasciolosis in vivo. [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]

[Display omitted] • Analysis of large, cross-regional data demonstrates the power of machine learning. • Dairy farms have distinct dissimilarities based on varying parasite exposure. • Relevant factors can remarkably differ across epidemiological settings. • Local and farm level conditions are paramount in dairy cow poly-parasitism. Fasciola hepatica and Ostertagia ostertagi are cattle parasites with worldwide relevance for economic outcome as well as animal health and welfare. The on-farm exposure of cattle to both parasites is a function of host-associated, intrinsic, as well as environmental and farm-specific, extrinsic, factors. Even though knowledge on the biology of both parasites exists, sophisticated and innovative modelling approaches can help to deepen our understanding of key aspects fostering the exposure of dairy cows to these pathogens. In the present study, multiple multinomial logistic regression models were fitted via neural networks to describe the differences among farms where cattle were not exposed to either F. hepatica or O. ostertagi , to one parasite, or to both, respectively. Farm-specific production and management characteristics were used as covariates to portray these differences. This elucidated inherent farm characteristics associated with parasite exposure. In both studied regions, pasture access for cows, farm-level milk yield, and lameness prevalence were identified as relevant factors. In region 'South', adherence to organic farming principles was a further covariate of importance. In region 'North', the prevalence of cows with a low body condition score, herd size, hock lesion prevalence, farm-level somatic cell count, and study year appeared to be of relevance. The present study broadens our understanding of the complex epidemiological scenarios that could predict differential farm-level parasite status. The analyses have revealed the importance of awareness of dissimilarities between farms in regard to the differential exposure to F. hepatica and O. ostertagi. This provides solid evidence that dynamics and relevant factors differ depending on whether or not cows are exposed to F. hepatica, O. ostertagi , or to both. [ABSTRACT FROM AUTHOR]

Background: We aimed to determine species of liver fluke that predominately cause fascioliasis in sheep, goats, and cattle in the Sulaymaniyah Province, Iraq using the molecular technique of DNA sequencing and restriction fragment length polymorphism (RFLP). Methods: The samples were collected from November 2021 to May 2022. The flukes were collected from infected livers of livestock at the slaughterhouse of Sulaymaniyah Governorate, Iraq. Overall, 205 flukes were collected from 56 hosts, cattle (n=22), sheep (n=28), and goats (n=6). The specific primers for FCOX1 and 28S rDNA gene amplification were used. The PCR products were subjected to restriction fragment polymorphism (RFLP) assay using Hpy188III and Dra II restriction enzymes, besides DNA sequencing. Results: The results showed the genetic polymorphisms among the flukes. Three patterns of RFLP were observed Fasciola hepatica, F. gigantica, and F. intermediate, where 28 of them displayed F. hepatica (sheep, n=14, goat, n=3 and cattle, n= 11), whereas 24 samples displayed the F. gigantica (sheep, n=12, goat, n=3 and cattle, n= 9), and only four samples belonged to F. intermediate (sheep n=3 and cattle, n=1). In addition, the result of the ribosomal DNA (28S rDNA) sequencing confirmed that the isolated flukes belonged to F. hepatica, F. gigantica and F. intermediate. Conclusion: All three main species are present in the study area and F. hepatica predominated among the animal species in this area also, our results concluded that PCR-RFLP is a rapid and reliable method for liver fluke species identification. [ABSTRACT FROM AUTHOR]

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The expression of the Fasciola hepatica carboxylesterase type B (CestB) gene is known to be induced upon exposure to the anthelmintic triclabendazole (TCBZ), leading to a substantial rise in enzyme-specific activity. Furthermore, the nucleotide sequence of the CestB gene displays variations that can potentially result in radical amino acid substitutions at the ligand binding site. These substitutions hold the potential to impact both the ligand–protein interaction and the catalytic properties of the enzyme. Thus, the objective of our study was to identify novel CestB polymorphisms in TCBZ-resistant parasites and field isolates obtained from a highly endemic region in Central Mexico. Additionally, we aimed to assess these amino acid polymorphisms using 3D modeling against the metabolically oxidized form of the anthelmintic TCBZSOX. Our goal was to observe the formation of TCBZSOX-specific binding pockets that might provide insights into the role of CestB in the mechanism of anthelmintic resistance. We identified polymorphisms in TCBZ-resistant parasites that exhibited three radical amino acid substitutions at positions 147, 215, and 263. These substitutions resulted in the formation of a TCBZSOX-affinity pocket with the potential to bind the anthelmintic drug. Furthermore, our 3D modeling analysis revealed that these amino acid substitutions also influenced the configuration of the CestB catalytic site, leading to alterations in the enzyme's interaction with chromogenic carboxylic ester substrates and potentially affecting its catalytic properties. However, it is important to note that the TCBZSOX-binding pocket, while significant for drug binding, was located separate from the enzyme's catalytic site, rendering enzymatic hydrolysis of TCBZSOX impossible. Nonetheless, the observed increased affinity for the anthelmintic may provide an explanation for a drug sequestration type of anthelmintic resistance. These findings lay the groundwork for the future development of a molecular diagnostic tool to identify anthelmintic resistance in F. hepatica. [ABSTRACT FROM AUTHOR]

Fasciolosis is an important economic disease of livestock. There is a global interest in the development of protective vaccines since the current anthelmintic therapy is no longer sustainable. A better knowledge of the host–parasite interaction is needed to design effective vaccines. To date, few studies have evaluated host–parasite interaction by comparing infected and reinfected animals. The present study evaluates the microscopical hepatic lesions in sheep infected and reinfected with Fasciola hepatica during the acute and chronic stages of infection. The histopathological study revealed the presence of necrotizing foci (NF1) associated with larvae migration during the early stages of infection in the primoinfected (PI) and reinfected (RI) groups. In the late stages of infection of the PI group and at the early and late stages of infection in the RI groups, extensive necrotizing/hemorrhagic foci (NF2) were found in the vicinity of enlarged bile ducts, some containing adult flukes, suggesting parasites may have caused NF2 while feeding. The immunohistochemical study revealed an increase in Foxp3+ T cells in both PI and RI groups with respect to the UC group and in the infiltrates adjacent to NF1 in the RI groups with respect to the PI group, suggesting the F. hepatica induce Foxp3 T cell expansion to facilitate parasite survival. In addition, in both the PI and RI groups, and during acute and chronic stages of the infection, a poor expression of iNOS was found accompanied by a strong expression of CD163, suggesting a marked M2 activation of macrophages in the hepatic lesions, which may be related with healing processes, and it also may facilitate parasite survival. The main differences between PI and RI animals were the more severe infiltration of eosinophils and Foxp3+ T cells, whereas RI did not modify M2 activation of macrophages which occurs since the early stages of primoinfection.

Fasciola hepatica has a complex lifecycle with multiple intermediate and definitive hosts and influenced by environmental factors. The disease causes significant morbidity in children and its prevalent worldwide. There is lack of data about distribution and burden of the disease in endemic regions, owing to poor efficacy of the different diagnostic methods used. A novel PCR-based test was developed by using a portable mini-PCR® platform to detect Fasciola sp. DNA and interpret the results via a fluorescence viewer and smartphone image analyzer application. Human stool, snail tissue, and water samples were used to extract DNA. Primers targeting the ITS-1 of the 18S rDNA gene of Fasciola sp. were used. The limit of detection of the mini-PCR test was 1 fg/μL for DNA samples diluted in water, 10 fg/μL for Fasciola/snail DNA scramble, and 100 fg/μL for Fasciola/stool DNA scramble. The product detection by agarose gel, direct visualization, and image analyses showed the same sensitivity. The Fh mini-PCR had a sensitivity and specificity equivalent to real-time PCR using the same specimens. Testing was also done on infected human stool and snail tissue successfully. These experiments demonstrated that Fh mini-PCR is as sensitive and specific as real time PCR but without the use of expensive equipment and laboratory facilities. Further testing of multiple specimens with natural infection will provide evidence for feasibility of deployment to resource constrained laboratories.

Fasciola hepatica is a trematode causing acute and chronic infection. A 33-year-old Canadian woman with eosinophilic liver abscesses and no relevant travel was diagnosed with F hepatica infection. F hepatica is reported in livestock in Alberta. This is the first case of locally acquired fascioliasis in Canada in >100 years. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is distributed worldwide, causing substantial economic losses in the animal husbandry industry. Human fasciolosis is an emerging zoonosis in Andean America, Asia, and Africa. The control of the disease, both in humans and animals, is based on using anthelmintic drugs, which has resulted in increased resistance to the most effective anthelmintics, such as triclabendazole, in many countries. This, together with the concerns about drug residues in food and the environment, has increased the interest in preventive measures such as a vaccine to help control the disease in endemic areas. Despite important efforts over the past two decades and the work carried out with numerous vaccine candidates, none of them has demonstrated consistent and reproducible protection in target species. This is at least in part due to the high immunomodulation capacity of the parasite, making ineffective the host response in susceptible species such as ruminants. It is widely accepted that a deeper knowledge of the host-parasite interactions is needed for a more rational design of vaccine candidates. In recent years, the use of emerging technologies has notably increased the amount of data about these interactions. In the present study, current knowledge of host-parasite interactions and their implication in Fasciola hepatica vaccine development is reviewed.

This study aimed to develop maps for Fasciola hepatica infection occurrence in dairy cattle in the districts of Matahuasi and Baños in the Peruvian central highlands. For this, a model based on the correlation between environmental variables and the prevalence of infection was constructed. Flukefinder® coprological test were performed in samples from dairy cattle from 8 herds, during both the rainy and wet season. Grazing plots were geo-referenced to obtain information on environmental variables. Monthly temperature, monthly rainfall, elevation, slope, normalized difference vegetation index (NDVI), enhanced vegetation index (EVI), normalized difference water index (NDWI), distance to rivers, urban areas and roads were obtained by using remote sensor images and ArcGIS®. Multilayer perceptron Artificial Neural Networks modeling were applied to construct a predictive model for the occurrence of fasciolosis, based on the relationship between environmental variables and level of infection. Kappa coefficient (k > 0.6) was used to evaluate concordance between observed and forecasted risk by the model. Coprological results demonstrated an average prevalence from 20% to 100%, in Matahuasi, and between 0 and 87.5%, in Baños. A model with a high level of concordance between predicted and observed infection risk (k = 0.77) was obtained, having as major predicting variables: slope, NDWI, NDVI and EVI. Fasciolosis risk was categorized as low (p

Background: We aimed to determine species of liver fluke that predominately cause fascioliasis in sheep, goats, and cattle in the Sulaymaniyah Province, Iraq using the molecular technique of DNA sequencing and restriction fragment length polymorphism (RFLP). Methods: The samples were collected from November 2021 to May 2022. The flukes were collected from infected livers of livestock at the slaughterhouse of Sulaymaniyah Governorate, Iraq. A total of 205 flukes were collected from 56 hosts, cattle (n=22), sheep (n=28), and goats (n=6). The specific primers for FCOX1 and 28S rDNA gene amplification were used. The PCR products were subjected to restriction fragment polymorphism (RFLP) assay using Hpy188III and Dra II restriction enzymes, besides DNA sequencing. Results: The results showed the genetic polymorphisms among the flukes. Three patterns of RFLP were observed Fasciola hepatica, F. gigantica, and F. intermediate, where 28 of them displayed F. hepatica (sheep, n=14, goat, n=3 and cattle, n= 11), whereas 24 samples displayed the F. gigantica (sheep, n=12, goat, n=3 and cattle, n= 9), and only four samples belonged to F. intermediate (sheep n=3 and cattle, n=1). In addition, the result of the ribosomal DNA (28S rDNA) sequencing confirmed that the isolated flukes belonged to F. hepatica, F. gigantica and F. intermediate. Conclusion: All three main species are present in the study area and F. hepatica predominated among the animal species in this area also, our results concluded that PCR-RFLP is a rapid and reliable method for liver fluke species identification.

Long non-coding (lnc)RNAs are a class of eukaryotic RNA that do not code for protein and are linked with transcriptional regulation, amongst a myriad of other functions. Using a custom in silico pipeline we have identified 6,436 putative lncRNA transcripts in the liver fluke parasite, Fasciola hepatica, none of which are conserved with those previously described from Schistosoma mansoni. F. hepatica lncRNAs were distinct from F. hepatica mRNAs in transcript length, coding probability, exon/intron composition, expression patterns, and genome distribution. RNA-Seq and digital droplet PCR measurements demonstrated developmentally regulated expression of lncRNAs between intra-mammalian life stages; a similar proportion of lncRNAs (14.2%) and mRNAs (12.8%) were differentially expressed (p<0.001), supporting a functional role for lncRNAs in F. hepatica life stages. While most lncRNAs (81%) were intergenic, we identified some that overlapped protein coding loci in antisense (13%) or intronic (6%) configurations. We found no unequivocal evidence for correlated developmental expression within positionally correlated lncRNA:mRNA pairs, but global co-expression analysis identified five lncRNA that were inversely co-regulated with 89 mRNAs, including a large number of functionally essential proteases. The presence of micro (mi)RNA binding sites in 3135 lncRNAs indicates the potential for miRNA-based post-transcriptional regulation of lncRNA, and/or their function as competing endogenous (ce)RNAs. The same annotation pipeline identified 24,141 putative lncRNAs in F. gigantica. This first description of lncRNAs in F. hepatica provides an avenue to future functional and comparative genomics studies that will provide a new perspective on a poorly understood aspect of parasite biology. Author summary: With increased understanding of genomes and transcriptomes it has become clear that not all RNAs code for protein, in contrast to the Central Dogma of molecular biology. Nevertheless, many such "non-coding" RNAs have functions in their own right. For example, long non-coding (lnc)RNAs comprise a large proportion of the human genome and control expression of some human protein coding genes. In parasites, lncRNAs remain understudied and poorly understood. This paper provides the first description of the lncRNA complement of the liver fluke, Fasciola hepatica. We show that lncRNA expression changes as parasites develop, in a similar manner to protein-coding mRNA, suggesting important roles in parasite growth and development. Some lncRNAs show correlated developmental expression with mRNAs coding for important parasite proteins, suggesting that they may interact, suggesting regulatory functions for fluke lncRNAs. A larger subset of lncRNA may interact with micro (mi)RNAs, another type of non-coding RNA that are important regulators of gene expression in eukaryotes. These interactions are, again, important additions to our understanding of fluke lncRNA biology. Our ultimate goal is to discover ways to inhibit these interactions that could be used for new medicines for fluke infections which are a global scourge on the farming industry. [ABSTRACT FROM AUTHOR]

Fasciolosis is an endemic zoonotic parasitic disease with significant impacts on human health and both animal health and production. Early post‐infection impacts on the host remain unclear. The objective of this study was to determine the changes, if any, to levels of endotoxin in cattle plasma in response to early‐stage infection with Fasciola hepatica. Thirty‐six (36) commercial bred cattle were experimentally infected with approximately 400 viable metacercariae. Plasma lipopolysaccharide (endotoxin) levels were examined on 24 occasions from 0 h before infection to 336 h after infection using the Limulus Amoebocyte Lysate chromogenic end point assay and compared with that of six (6) uninfected control animals. Peak lipopolysaccharide levels in infected animals were reached at 52 h after infection and returned to pre‐infection levels at time 144 h after infection. Infected animals had significantly elevated lipopolysaccharide levels between 24 and 120 h after infection when compared to uninfected animals. The mean change in endotoxin units (EU)/mL over time after infection was statistically significant in infected animals. Elevations of lipopolysaccharide occurred in all infected animals suggesting a possible repeatable and titratable endotoxemia conducive to therapeutic agent model development. [ABSTRACT FROM AUTHOR]

The Crucial Role of Raw Green Vegetables in Nourishing Well-being. However, lurking within these natural wonders lies an inconspicuous peril - the potential conduit for intestinal parasites due to potential parasitological contamination. Our ongoing research endeavor is laser-focused on scrutinizing the prevalence of parasitic protozoa and helminths' cysts and eggs within the raw vegetables available in the bustling retail markets of Al-Amarah City and the serene Qal'at Saleh District, both nestled in the heart of Maysan Province, Southern Iraq. During our investigation, we meticulously scrutinized 180 samples representing a diverse array of five vegetable varieties, encompassing radish, leek, cress, celery, and lettuce. Utilizing established and recognized testing protocols, we rigorously conducted our examinations. The results illuminated a concerning reality: among the fresh leafy vegetables surveyed in Al-Amarah City, 17.76% bore the burden of contamination, while in the picturesque Qal'at Saleh District, this figure ascended to a troubling 22.2%. Within these innocent greens lay the insidious presence of helminth eggs and parasitic protozoan cysts, a revelation demanding our attention and action. Entamoeba histolytica and Giardia lamblia cysts were detected at 8.88% and 1.11% respectively in Al-Amarah City and at 6.66% and 2.22% in Qal'at Saleh District. On the contrary, Eggs of Hymenolepis nana and Fasciola hepatica were detected at 2.22% and 1.11% respectively in Al-Amarah City and at 5.55% and 2.22% in Qal'at Saleh District. Eggs of Ascaris lumbricoides were detected at 4.44% and 5.55% in Al-Amarah City and Qal'at Saleh District respectively. Lettuce showed the highest contamination percentage in comparison with other sorts of vegetables at 27.77% in Al-Amarah City and 33.33% in Qal'at Saleh District with Noteworthy Disparities Evident at a Significance Threshold of P<0.05: This investigation has reached a decisive verdict, underscoring the latent peril residing within the raw vegetables retailed in Maysan Province's local markets. These innocent greens, it transpires, harbor the potential to serve as conduits for intestinal protozoa and helminths, thereby casting a shadow over the health of those who partake in such produce. The author advocates a compelling case for the imperative involvement of pertinent authorities. In doing so, the endeavor should revolve around mitigating the extent of parasitic pathogen contamination within plants, a mission that hinges on the twin pillars of vendor education and community awareness. [ABSTRACT FROM AUTHOR]

Fasciola hepatica is a trematode leading to heavy economic setbacks to the livestock sector globally. The population's genetic information and intimate kinship level are frequently assessed using analysis of mitochondrial DNA. In this analysis, we retrieved cox1 (n = 247) and nad1 (n = 357) sequences of F. hepatica from the NCBI GenBank database and aligned the sequences with the respective reference sequences using MEGA software. The median joining network was drawn using PopArt software while neutrality and diversity indices were estimated with the help of DnaSp software. Neighbor-joining phylogenetic tree was constructed using the MEGA software package. A total of 46 and 98 distinctive haplotypes were observed for cox1 and nad1 genes, respectively. Diversity indices indicated high haplotype and nucleotide diversities in both genes. Positive Tajima's D and Fu's Fs values were found for the entire population of both the genes under study. The cox1 and nad1 gene segments in this study showed high Tajima's D values, suggesting a low likelihood of future population growth. The Tajima's D value of the nad1 gene sequence is lower (2.14910) than that of the cox1 gene sequence (3.40314), which suggests that the former is growing at a slower rate. However, the region-wise analysis revealed that both the cox1 and nad1 genes showed deviation from neutrality suggesting a recent population expansion as a result of an excess of low-frequency polymorphism. Furthermore, the overall host-wise analysis showed positive and significant Tajima's D values for the cox1 and nad1 gene sequences. To the best of our knowledge, this is the first attempt to provide insights into genetic variations and population structure of F. hepatica at a global scale using cox1 and nad1 genes. Our findings suggest the existence of specific variants of F. hepatica in different parts of the world and provide information on the molecular ecology of F. hepatica. The results of this study also mark a critical development in upcoming epidemiological investigations on F. hepatica and will also contribute to understanding the global molecular epidemiology and population structure of F. hepatica. [ABSTRACT FROM AUTHOR]

Key Clinical message: Fascioliasis poses diagnostic challenges to clinicians for its broad spectrum of hepatobiliary symptoms and lower detection rates. Timely and precise identification avoids long‐standing hepatic complications. Fasciola hepatica, a trematode parasite, inhabits snails and sheep. Human hepatic fascioliasis is a neglected tropical disease with no specific tests. In this instance, the significance of clinical awareness, appropriate imaging, and serological investigations is demonstrated. A young Nepalese girl was diagnosed and managed successfully. [ABSTRACT FROM AUTHOR]

Fascioliasis is a worldwide spread parasitic disease with a high impact on the economy and public health in many countries. Therefore, it is important to apply prevention, effective treatment, and education campaigns, to prevent the disease. This work aimed to determine the biochemical markers of liver damage caused by Fasciola hepatica during 21 days (D0 – D21) post albendazole (ABZ) therapy in the blood sera. This study included 24 cows naturally infected with Fasciola hepatica, randomly divided into a control group (n = 12) and a group treated per os with ABZ (7.5 mg per kg body weight) (n = 12). The activity of hepatic enzymes – aspartate aminotransferase (AST), alanine aminotransferase (ALT) and cholestatic enzymes – gamma-glutamyl transferase (GGT), and alkaline phosphatase (ALP), were measured in both groups. The activity of AST has significantly increased in the course of the experiment (D0 – D21) in both groups (P < 0.001). ALT activity during D0 – D21 was significantly decreased in both groups (P < 0.001). Activity of GGT was not significantly changed in the control group in the course of the experiment (P > 0.05). Contrarily, the activity of GGT in the treated group was significantly decreased (P < 0.01) during D0 – D21. The activity of GGT in the blood sera of the treated group was significantly lower in comparison with the control group in D7 and D21 (P < 0.01). This decreased GGT activity in the blood sera of the treated group may be caused by the effect of albendazole resulting in fasciola elimination and bile flow improvement. The results of ALP determination show no significant changes in ALP activity between groups. This can lead to the conclusion that the most useful marker of cholestasis caused by F. hepatica is the GGT activity and may help to evaluate the ABZ therapy effect in cattle. [ABSTRACT FROM AUTHOR]

Purpose: Fasciola hepatica is a globally distributed trematode that causes significant economic losses. Triclabendazole is the primary pharmacological treatment for this parasite. However, the increasing resistance to triclabendazole limits its efficacy. Previous pharmacodynamics studies suggested that triclabendazole acts by interacting mainly with the β monomer of tubulin. Methods: We used a high-quality method to model the six isotypes of F. hepatica β-tubulin in the absence of three-dimensional structures. Molecular dockings were conducted to evaluate the destabilization regions in the molecule against the ligands triclabendazole, triclabendazole sulphoxide and triclabendazole sulphone. Results: The nucleotide binding site demonstrates higher affinity than the binding sites of colchicine, albendazole, the T7 loop and pβVII (p < 0.05). We suggest that the binding of the ligands to the polymerization site of β-tubulin can lead a microtubule disruption. Furthermore, we found that triclabendazole sulphone exhibited significantly higher binding affinity than other ligands (p < 0.05) across all isotypes of β-tubulin. Conclusions: Our investigation has yielded new insight on the mechanism of action of triclabendazole and its sulphometabolites on F. hepatica β-tubulin through computational tools. These findings have significant implications for ongoing scientific research ongoing towards the discovery of novel therapeutics to treat F. hepatica infections. [ABSTRACT FROM AUTHOR]

Background: Fasciolosis (Fasciola hepatica) and paramphistomosis (Calicophoron daubneyi) are two important infections of livestock. Calicophoron daubneyi is the predominant Paramphistomidae species in Europe, and its prevalence has increased in the last 10–15 years. In Italy, evidence suggests that the prevalence of F. hepatica in ruminants is low in the southern part, but C. daubneyi has been recently reported at high prevalence in the same area. Given the importance of reliable tools for liver and rumen fluke diagnosis in ruminants, this study evaluated the diagnostic performance of the Mini-FLOTAC (MF), Flukefinder® (FF) and sedimentation (SED) techniques to detect and quantify F. hepatica and C. daubneyi eggs using spiked and naturally infected cattle faecal samples. Methods: Briefly, negative bovine faecal samples were artificially spiked with either F. hepatica or C. daubneyi eggs to achieve different egg count levels: 10, 50 and 100 eggs per gram (EPG) of faeces. Moreover, ten naturally infected cattle farms from southern Italy with either F. hepatica and/or C. daubneyi were selected. For each farm, the samples were analysed individually only with MF technique and as pools using MF, FF and SED techniques. Bayesian latent class analysis (LCA) was used to estimate sensitivity and accuracy of the predicted intensity of infection as well as the infection rate in the naturally infected farms. Results: The outcome of this study showed that the highest number of eggs (F. hepatica and C. daubneyi) recovered was obtained with MF, followed by FF and SED in spiked infected samples at 50 and 100 EPG, while at lower infection levels of 10 EPG, FF gave the best results. Moreover, the sensitivity for all the techniques included in the study was estimated at > 90% at infection levels > 20 EPG for both F. hepatica and C. daubneyi eggs. However, MF was the most accurate of the three techniques evaluated to estimate fluke infection intensity. Nevertheless, all three techniques can potentially estimate infection rate at farm level accurately. Conclusions: Optimization and standardization of techniques are needed to improve the FEC of fluke eggs. [ABSTRACT FROM AUTHOR]

Diabetes is the fastest growing chronic disease globally, with prevalence increasing at a faster rate than heart disease and cancer. While the disease presents clinically as chronic hyperglycaemia, two distinct subtypes have been recognised. Type 1 diabetes (T1D) is characterised as an autoimmune disease in which the insulin-producing pancreatic b-cells are destroyed, and type 2 diabetes (T2D) arises due to metabolic insufficiency, in which inadequate amounts of insulin are produced, and/or the actions of insulin are diminished. It is now apparent that pro-inflammatory responses cause a loss of functional bcell mass, and this is the common underlying mechanism of both T1D and T2D. Macrophages are the central immune cells in the pathogenesis of both diseases and play a major role in the initiation and perpetuation of the proinflammatory responses that compromise b-cell function. Furthermore, it is the crosstalk between macrophages and b-cells that orchestrates the inflammatory response and ensuing b-cell dysfunction/destruction. Conversely, this crosstalk can induce immune tolerance and preservation of b-cell mass and function. Thus, specifically targeting the intercellular communication between macrophages and b-cells offers a unique strategy to prevent/halt the islet inflammatory events underpinning T1D and T2D. Due to their potent ability to regulate mammalian immune responses, parasitic worms (helminths), and their excretory/secretory products, have been examined for their potential as therapeutic agents for both T1D and T2D. This research has yielded positive results in disease prevention, both clinically and in animal models. However, the focus of research has been on the modulation of immune cells and their effectors. This approach has ignored the direct effects of helminths and their products on b-cells, and the modulation of signal exchange between macrophages and b-cells. This review explores how the alterations to macrophages induced by helminths, and their products, influence the crosstalk with b-cells to promote their function and survival. In addition, the evidence that parasite-derived products interact directly with endocrine cells to influence their communication with macrophages to prevent b-cell death and enhance function is discussed. This new paradigm of two-way metabolic conversations between endocrine cells and macrophages opens new avenues for the treatment of immune-mediated metabolic disease. [ABSTRACT FROM AUTHOR]

In fascioliasis, oligoxenous snail vector specificity, low mammal host specificity, epidemiological situations, transmission patterns, and influencing biotic/abiotic factors underlie high complexity. In human fascioliasis, a One Health approach is the best strategy to complement preventive chemotherapy by decreasing infection/reinfection risk. A One Health multidisciplinarity approach for human fascioliasis includes axes targeting the environment (habitat and climate), snail vectors, mammal reservoirs, the human host, and ethnography of the inhabitants of the endemic area. Sufficient baseline knowledge about epidemiology and transmission of fascioliasis obtained in previous field surveys and experimental studies is needed. A One Health action should be adapted to the epidemiological and transmission characteristics of the endemic area, and use a pilot area when dealing with large endemic areas. Fasciola hepatica and F. gigantica are liver flukes causing fascioliasis, a worldwide zoonotic, complex disease. Human infection/reinfection occurs in endemic areas where preventive chemotherapy is applied, because of fasciolid transmission ensured by livestock and lymnaeid snail vectors. A One Health control action is the best complement to decrease infection risk. The multidisciplinary framework needs to focus on freshwater transmission foci and their environment, lymnaeids, mammal reservoirs, and inhabitant infection, ethnography and housing. Local epidemiological and transmission knowledge furnished by previous field and experimental research offers the baseline for control design. A One Health intervention should be adapted to the endemic area characteristics. Long-term control sustainability may be achieved by prioritizing measures according to impact depending on available funds. [ABSTRACT FROM AUTHOR]

Parasites are a significant component of biodiversity. They negatively affect fish appearance, growth, and reproduction. In this study, the prevalence of infection, diversity, and mean intensity of parasites were examined in 9 freshwater fish species (45 samples per fish species). Ecto-parasites were examined on the skin, gills, and fins with a hand lens. Wet mounts were prepared using mucosal scrapings from all the external and internal organs of the sampled fish. Microscopy, muscle compression, and the pepsin-HCL artificial digestion technique were also performed. In this study, 26 species of parasites were identified including three taxa belonging to 9 species of protozoan parasites, 11 treamtodes, and 6 monogenean parasites. The identified protozoan parasites were Entamoeba histolitica, Chilodonella sp., Coccidia sp., Costia sp., Cryptobia sp., Ichthyopthiris-multifilis, Microsporidia, Piscinoodinium sp., and Ichthyobodo necator. The identified trematode parasites were Fasciola gigantica, Echinostoma revolutum, Fasciola hepatica, Haplorchis pumilio, Brachylaima cribbi, Echinostoma cinetorchis, Neascus sp., Deropegus sp., Trematode Soldier, Centrocestus formosanus, and Clinostomum marginatum. The identified monogenean parasites were Dactylogyrus limipopoensis, Dactylogyrus anchoratus, Dactylogyrus myersi, Dactylogyrus vastator, Gyrodactylus salaris, and Ancyrocephalus. The diversity of parasites was maximum at the Okara site. The host's organs that were targeted for parasitic infection included the intestine, liver, gills, fins, skin, and kidneys. The majority of the parasites were identified in Labeo rohita followed by Hypophthalmichthys molitrix, Ctenopharyngodon idella, Oreochromis niloticus, Cyprinus carpio, and Wallagu attu. Two species appeared to be resistant species because none of the parasites were observed in Notopterus notopterus or Sperata seenghala. This study also concluded that the prevalence of parasites increased with increasing length, size, and age of fish. [ABSTRACT FROM AUTHOR]

SARS-CoV-2, the causal agent of COVID-19, is a new coronavirus that has rapidly spread worldwide and significantly impacted human health by causing a severe acute respiratory syndrome boosted by a pulmonary hyperinflammatory response. Previous data from our lab showed that the newly excysted juveniles of the helminth parasite Fasciola hepatica (FhNEJ) modulate molecular routes within host cells related to vesicle-mediated transport and components of the innate immune response, which could potentially be relevant during viral infections. Therefore, the aim of the present study was to determine whether FhNEJ-derived molecules influence SARS-CoV-2 infection efficiency in Vero cells. Pre-treatment of Vero cells with a tegument-enriched antigenic extract of FhNEJ (FhNEJ-TEG) significantly reduced infection by both vesicular stomatitis virus particles pseudotyped with the SARS-CoV-2 Spike protein (VSV-S2) and live SARS-CoV-2. Pre-treatment of the virus itself with FhNEJ-TEG prior to infection also resulted in reduced infection efficiency similar to that obtained by remdesivir pre-treatment. Remarkably, treatment of Vero cells with FhNEJ-TEG after VSV-S2 entry also resulted in reduced infection efficiency, suggesting that FhNEJ-TEG may also affect post-entry steps of the VSV replication cycle. Altogether, our results could potentially encourage the production of FhNEJ-derived molecules in a safe, synthetic format for their application as therapeutic agents against SARS-CoV-2 and other related respiratory viruses. [ABSTRACT FROM AUTHOR]

Infection with Fasciola hepatica (liver fluke) causes fasciolosis (or fascioliasis) and poses a considerable economic as well as welfare burden to both the agricultural and animal health sectors. Here, we explore the ex vivo anthelmintic potential of synthetic derivatives of hederagenin, isolated in bulk from Hedera helix. Thirty-six compounds were initially screened against F. hepatica newly excysted juveniles (NEJs) of the Italian strain. Eleven of these compounds were active against NEJs and were selected for further study, using adult F. hepatica derived from a local abattoir (provenance unknown). From these eleven compounds, six demonstrated activity and were further assessed against immature liver flukes of the Italian strain. Subsequently, the most active compounds (n = 5) were further evaluated in ex vivo dose response experiments against adult Italian strain liver flukes. Overall, MC042 was identified as the most active molecule and the EC50 obtained from immature and adult liver fluke assays (at 24 h post co-culture) are estimated as 1.07 μM and 13.02 μM, respectively. When compared to the in vitro cytotoxicity of MDBK bovine cell line, MC042 demonstrated the highest anthelmintic selectivity (44.37 for immature and 3.64 for adult flukes). These data indicate that modified hederagenins display properties suitable for further investigations as candidate flukicides. [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]

Fasciola species are a member of flatworms belonging to the trematodes (flukes), commonly known as liver fluke, they are extremely pathogenic parasites that affect the liver of humans and animals, nowadays, most laboratories and research facilities use molecular-based techniques for identifying and describing Fasciola species. The molecular diagnostic markers such as polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and PCR-RFLP methods are accurate and more specific than the immunological and microscopical methods. The identification of the species of liver flukes will give a new clue for the treatment and control of fascioliasis. The aim, of this study, is to find the molecular characterization of Fasciola spp. isolated from humans and animals in Sulaimani city. The flukes were isolated from humans using endoscopic techniques and from slaughtered livestock at the new slaughterhouse of Sulaimani, 48 liver flukes were collected from different hosts; human (n = 3), cattle (n = 20), sheep (n = 20), and goats (n = 5) from October 2021 to April 2022. The uinversal primers ribosomal Deoxy Ribo Nuclic Acid (rDNA) were used, then the PCR products were subjected to restriction fragment polymorphism (RFLP) assay and The PCR Product was digested with restriction enzymes DraII, also the DNA sequencing was used for the PCR product of the primer Cytochrome Oxidase subuint 1 (COX1). The results of the PCR-RFLP of the 28s rDNA show the genetic polymorphisms among the flukes and two patterns of RFLP were observed F. hepatica, and F. gigantica, also the sequence analysis of the partial gene of the COX1 showed the isolated flukes belonged to F. hepatica and F. gigantica with some genetic variation, and the result of the sequences was deposited in the Gene Bank under the following Accession numbers; F. gigantica (OP718780 and OP718781) and F. hepatica (OP718782, OP718783, and OP718784). The present study concludes that F. hepatica and F. gigantica are both responsible for human and animal Fasciolasis in Kurdistan-Iraq, Therefore, RFLP techniques and DNA sequencing are a reliable, and differential method for species and genotyping identification of liver fluke.

Background and Aim: Extensive cattle rearing is a major source of economy for the inhabitants of the Amazon region of Peru. Milk and meat production is generally affected by the prevalence of various parasites, including hepatic and gastrointestinal parasites, as these products provide ideal conditions for parasitic growth. This poses a serious public health threat. This study aimed to estimate the prevalence, coinfection, and risk factors associated with the liver fluke (Fasciola hepatica) and other gastrointestinal parasites in cattle from the Amazon region of Peru. Materials and Methods: Fecal samples obtained from 1450 bovine specimens were analyzed using flotation and sedimentation methods to identify parasites, including Eimeria spp., strongyle-type eggs (STEs), and F. hepatica. We collected information about the specimens, including age, sex, origin, breed, category, frequency of deworming, farm size, herd size, water sources, and rearing system by conducting simple inspections and interviewing owners. The data obtained were statistically evaluated using the Chi-square test (p < 0.05) to determine the association between the qualitative variables. We also calculated the odds ratio at a 95% confidence interval to identify the risk factors. Results: We observed that F. hepatica, Eimeria spp., and STEs were 45.6%, 39.8%, and 35.3% prevalent, respectively. We found risk factors related to distomatosis in the animals from Huambo, where the drinking water sources are mainly streams, ditches, and rivers, while the specimens from Valle Chico were predisposed to coccidiosis. Further, the risk factors related to the presence of STEs in feces were age (61–90 months), origin (Valle Chico), herd size (

Abstract 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.

Objective: Lymnaea stagnalis known as the great pond snail, is one of the intermediate hosts of Fasciola hepatica, a zoonotic parasite. In this study, it was aimed to determine the larval forms of F. hepatica by polymerase chain reaction (PCR) in L. stagnalis species snails collected from the vicinity of Ağrı province. Methods: In this study, 150 L. stagnalis snails were collected from the Ağrı province. The freshwater snails brought to the laboratory were dissected, then their soft tissues were examined under a microscope. DNA extraction was performed on the dissected snails. After DNA extraction, PCR was performed using primers targeting the cytochrome c oxidase subunit 1 gene region. Results: In the microscopic examination, larval forms of F. hepatica could not be detected. However, it was concluded that two (1.3%) L. stagnalis freshwater snails were infected with the larval forms of F. hepatica in the PCR. Conclusion: It was determined that L. stagnalis served as an intermediate host to F. hepatica in the study area.

Rumen fluke (RF) are trematode parasites that have increased in prevalence within European ruminant livestock since the 1990s. Morbidity and mortality can result from the development of juvenile flukes in the duodenum, however, evidence for significant impacts of adult fluke in the rumen of the final host is equivocal. The presence of rumen fluke in Scotland had not previously been quantified, so the purpose of this study was to use historical coprological data collected between 2008 and 2018 at the School of Veterinary Medicine in Glasgow to evaluate the frequency of rumen fluke infection and to compare this with the presence of liver fluke. This analysis showed evidence of adult rumen fluke in 6.6% of samples submitted, with a substantial and significant increase in positive diagnoses from 2016, following which positive rumen fluke diagnoses equaled or slightly outnumbered those of liver fluke. A prospective post-mortem examination study was also carried out to determine the presence of rumen and liver fluke in cattle and sheep, to quantify adult rumen fluke burdens and to assess any pathological changes in the reticulorumen and proximal duodenum associated with infection. The presence of rumen fluke in post-mortem cases was 26.9% (n = 18/67), the majority (66.7%, n = 12/18) with burdens of less than 100 adult rumen flukes. There was no significant difference in mean ruminal papillar density and length in animals with and without adult rumen fluke and no significant gross pathology was observed in the rumen or reticulum. Examination of animals with adult rumen fluke provided no evidence of any consistent associations with acute or chronic inflammatory changes in the duodenum. All rumen fluke collected at necropsy were identified by PCR and sequencing as Calicophoron daubneyi.

INTRODUCTION: We aimed to raise awareness for Fasciola hepatica infestation to be included in the differential diagnosis by revealing the clinical, laboratory and radiological findings and patient presentation. METHODS: Demographic characteristics, clinical and laboratory findings of 31 patients aged 18-90 years with Fasciola Hepatica infestation diagnosed by the Department of Infectious Diseases and Clinical Microbiology between 01.01.2013 and 05.06.2022 were retrospectively analyzed. RESULTS: Of the 31 cases, 21 (67,7%) were female, 10 (32,3%) were male, and the mean age was (40,6 +-21,1) years. All of the cases had abdominal pain complaints. When laboratory findings were examined, eosinophilia 29 (93,5%) was the most common abnormal finding. Fascioliasis suspicious lesion was observed in all cases by imaging methods. DISCUSSION AND CONCLUSION: The mean age of the cases in our study was 40,6 +-21,1 years, which was consistent with the literature. There was a history of living in rural areas in 20 (64,5%) cases. Imaging and laboratory findings were used for diagnosis. In our study, most of the cases were from the province of Van. Since watercress plant consumption is common in this region, it is thought that fascioliasis may be endemic in and around Van province. Living in this region; In cases with abdominal pain, weakness and eosinophilia, fascioliasis should be considered in the differential diagnosis.

Abstract The expression of proinflammatory (IL-1β, IFN-γ, TNF-α) and regulatory (IL-10, TGF-β, IL-4) cytokines, as well as the transcription factor FoxP3, was quantified in the liver and hepatic lymph node (HLN) of sheep primoinfected and reinfected with Fasciola hepatica at early (4, 8 and 16 days post-infection [dpi]) and late (100 dpi) stages. The liver exerted a Th2 immune response at very early stages after the primoinfection with F. hepatica that induced the downregulation of IFN-γ, followed by a Th1/Th2/Treg response although the late stages were characterised by the expression of Th1/Th2 immune mediators. Contrarily, in reinfected sheep a robust mixed Th1/Th2/Treg immune response was found at very early stages meanwhile at late stages we observed a Th2/Treg immune response overcoming the expression of Th1 immune mediators. However, the HLN displayed a completely different Th1/Th2/Treg expression profile compared to the liver. Primoinfections with F. hepatica in HLN induced a mixed Th1/Th2/Treg environment from early stages, establishing a Th2 immune response at a late stage. However, the reinfected sheep exerted a Th2 immune response at early stages led by the IL-4 expression in opposition to the Th1/Th2/Treg found in the liver, meanwhile at late stages the HLN of reinfected sheep exerted a mixed Th1/Th2/Treg immune response. This is the first work publishing the expression of immune mediators in the liver and HLN from reinfected sheep with F. hepatica. The study of the immune responses exerted by the natural host in the target organs directly implied in the development of F. hepatica are crucial to better understand the immunopathogenesis of the fasciolosis being a key factor to develop effective vaccines.

Abstract Background Glutamate carboxypeptidase 2 (GCP2) belongs to the M28B metalloprotease subfamily encompassing a variety of zinc-dependent exopeptidases that can be found in many eukaryotes, including unicellular organisms. Limited information exists on the physiological functions of GCP2 orthologs in mammalian tissues outside of the brain and intestine, and such data are completely absent for non-mammalian species. Here, we investigate GCP2 orthologs found in trematodes, not only as putative instrumental molecules for defining their basal function(s) but also as drug targets. Methods Identified genes encoding M28B proteases Schistosoma mansoni and Fasciola hepatica genomes were analyzed and annotated. Homology modeling was used to create three-dimensional models of SmM28B and FhM28B proteins using published X-ray structures as the template. For S. mansoni, RT-qPCR was used to evaluate gene expression profiles, and, by RNAi, we exploited the possible impact of knockdown on the viability of worms. Enzymes from both parasite species were cloned for recombinant expression. Polyclonal antibodies raised against purified recombinant enzymes and RNA probes were used for localization studies in both parasite species. Results Single genes encoding M28B metalloproteases were identified in the genomes of S. mansoni and F. hepatica. Homology models revealed the conserved three-dimensional fold as well as the organization of the di-zinc active site. Putative peptidase activities of purified recombinant proteins were assayed using peptidic libraries, yet no specific substrate was identified, pointing towards the likely stringent substrate specificity of the enzymes. The orthologs were found to be localized in reproductive, digestive, nervous, and sensory organs as well as parenchymal cells. Knockdown of gene expression by RNAi silencing revealed that the genes studied were non-essential for trematode survival under laboratory conditions, reflecting similar findings for GCP2 KO mice. Conclusions Our study offers the first insight to our knowledge into M28B protease orthologs found in trematodes. Conservation of their three-dimensional structure, as well as tissue expression pattern, suggests that trematode GCP2 orthologs may have functions similar to their mammalian counterparts and can thus serve as valuable models for future studies aimed at clarifying the physiological role(s) of GCP2 and related subfamily proteases. Graphical Abstract

Abstract Background Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial inflammation, fibroblast-like synoviocytes (FLS) activation and joint destruction. Fasciola hepatica is a platyhelminth that releases excretory-secretory immunomodulatory products capable of suppressing the Th1 immune response. Despite the effectiveness of available treatments for inducing disease remission, current options are not successful in all patients and may cause side effects. Thus, we evaluated the therapeutic potential of F. hepatica extract on FLS from RA patients and arthritis models. Methods FLS were isolated from synovial fluid of RA patients, cultured, and exposed to F. hepatica extract (60, 80, and 100 µg/ml) for different time points to assess cell viability, adherence, migration and invasion. For in vivo experiments, mice with antigen (AIA) and collagen (CIA) induced arthritis received a 200 µg/dose of F. hepatica extract daily. Statistical analysis was performed by ANOVA and Student's t-test using GraphPad Prism 6.0. Results In vitro assays showed that extract decreased FLS cell viability at concentration of 100 µg/ml (83.8% ± 5.0 extract vs. 100.0% ± 0.0 control; p

Background: Fasciolosis is a parasitic infection caused by the liver fluke Fasciola hepatica that can have a major economic impact on livestock industry. The prevalence of the disease has recently been increasing in many North European countries. The objective of this study was to determine the prevalence of antibody against F. hepatica in Finnish cattle herds and sheep flocks during 2019 by using a commercial enzyme-linked immunosorbent assay (ELISA). Randomly selected bulk tank milk samples were obtained from 660 dairy herds. Blood samples were collected at slaughterhouses from 1944 suckler cows from 309 herds and from 1120 sheep from 95 flocks. Results: Antibodies against F. hepatica were found in 0.45% (95% confidence interval (CI): 0.15–1.33) of the dairy herds and 0.97% (95% CI: 0.33–2.82) of the suckler cow herds. The seropositive herds were located in eastern and central Finland. None of the sampled sheep flocks tested had antibodies against F. hepatica (95% CI: 0–3.89). The results of the assays were compared with meat inspection data received from the slaughterhouses. All positive herds also had liver condemnations due to F. hepatica based on the meat inspection reports. Conclusions: Compared to other North European countries, the prevalence of fasciolosis in Finland can be considered low, and according to meat inspection reports, there are no indications of the prevalence increasing in Finland. [ABSTRACT FROM AUTHOR]

Background and Aim: Bovine fasciolosis is a reemerging neglected disease with a worldwide distribution caused by the trematode Fasciola spp., which parasitize various hosts. Bovine fasciolosis is responsible for large economic losses in the bovine livestock sector. This study aimed to estimate the seroprevalence and risk factors of bovine fasciolosis in the municipalities of Colombia. Materials and Methods: This was a descriptive cross-sectional study with simple random sampling conducted on 1140 cattle from the municipalities of Chiquinquirá, San Miguel de Sema, and Ubaté for a duration of 3 months. Serum samples were processed using the commercial Fasciola hepatica Antibody Test Kit IDEXX® Fasciolosis Verification (IDEXX, United States), which identified immunoglobulin G antibodies for gf2 antigen purified from Fasciola extracts. The f2 antigen is extremely immunogenic and highly specific for F. hepatica. An epidemiological survey was performed to record variables related to the sampled animals and herd management practices. Data were processed using the statistical program Epi Info® (Centers for Disease Control and Prevention; Atlanta, Georgia). The prevalence ratio was estimated to evaluate the association between fasciolosis and the hypothesized causal factors and the significance of this association using Pearson's Chi-square test. Finally, a logistic regression model was developed. Results: The overall seroprevalence was 72.3%. The seroprevalence was 83.9% (323/385) in Chiquinquirá, 68.17% (257/377) in Ubaté, and 64.55% (244/378) in San Miguel de Sema. The seroprevalence was higher in male animals in Chiquinquirá and in female animals in San Miguel de Sema and Ubaté. Similarly, sex showed a statistically significant association with disease prevalence in Ubaté. The highest prevalence was found in cattle aged >2 years. The Holstein breed showed maximum seroprevalence in Chiquinquirá (p = 0.05) and San Miguel de Sema, whereas crossbreed showed higher seroprevalence in Ubaté. Similarly, in Chiquinquirá, the association between the seroprevalence of fasciolosis and the presence of other species was statistically significant (95% confidence interval [CI]: 0.9601-3.4944; p = 0.0448). In Ubaté, the disease presentation was also associated with pasture rental (95% CI: 0.4047-1.0023; p = 0.003) and attendance to livestock expositions (95% CI: 0.2313-1.0636; p = 0.044). However, in San Miguel de Sema, water from the stream showed a statistically significant association with disease presentation (95% CI: 0.5209-1.0985; p = 0.00649785). Female sex and diarrhea occurrence were considered risk factors for fasciolosis. Conclusion: A high seroprevalence of antibodies to Fasciola spp. was detected in cattle in the study municipalities, indicating a high parasite distribution in these areas. Female sex and diarrhea were established as risk factors associated with fasciolosis in Ubaté and San Miguel de Sema, respectively. Further, research is necessary to establish prevention and control programs against parasitosis. [ABSTRACT FROM AUTHOR]

Fasciola species are a member of flatworms belonging to the trematodes (flukes), commonly known as liver fluke, they are extremely pathogenic parasites that affect the liver of humans and animals, nowadays, most laboratories and research facilities use molecular-based techniques for identifying and describing Fasciola species. The molecular diagnostic markers such as polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and PCR-RFLP methods are accurate and more specific than the immunological and microscopical methods. The identification of the species of liver flukes will give a new clue for the treatment and control of fascioliasis. The aim, of this study, is to find the molecular characterization of Fasciola spp. isolated from humans and animals in Sulaimani city. The flukes were isolated from humans using endoscopic techniques and from slaughtered livestock at the new slaughterhouse of Sulaimani, 48 liver flukes were collected from different hosts; human (n = 3), cattle (n = 20), sheep (n = 20), and goats (n = 5) from October 2021 to April 2022. The uinversal primers ribosomal Deoxy Ribo Nuclic Acid (rDNA) were used, then the PCR products were subjected to restriction fragment polymorphism (RFLP) assay and The PCR Product was digested with restriction enzymes DraII, also the DNA sequencing was used for the PCR product of the primer Cytochrome Oxidase subuint 1 (COX1). The results of the PCR-RFLP of the 28s rDNA show the genetic polymorphisms among the flukes and two patterns of RFLP were observed F. hepatica, and F. gigantica, also the sequence analysis of the partial gene of the COX1 showed the isolated flukes belonged to F. hepatica and F. gigantica with some genetic variation, and the result of the sequences was deposited in the Gene Bank under the following Accession numbers; F. gigantica (OP718780 and OP718781) and F. hepatica (OP718782, OP718783, and OP718784). The present study concludes that F. hepatica and F. gigantica are both responsible for human and animal Fasciolasis in Kurdistan-Iraq, Therefore, RFLP techniques and DNA sequencing are a reliable, and differential method for species and genotyping identification of liver fluke. [ABSTRACT FROM AUTHOR]

Context: Quinoxaline 1,4-di-N-oxide is a scaffold with a wide array of biological activities, particularly its use to develop new antiparasitic agents. Recently, these compounds have been described as trypanothione reductase (TR), triosephosphate isomerase (TIM), and cathepsin-L (CatL) inhibitors from Trypanosoma cruzi, Trichomonas vaginalis, and Fasciola hepatica, respectively. Methods: Therefore, the main objective of this work was to analyze quinoxaline 1,4-di-N-oxide derivatives of two databases (ZINC15 and PubChem) and literature by molecular docking, dynamic simulation and complemented by MMPBSA, and contact analysis of molecular dynamics' trajectory on the active site of the enzymes to know their potential effect inhibitory. Interestingly, compounds Lit_C777 and Zn_C38 show preference as potential TcTR inhibitors over HsGR, with favorable energy contributions from residues including Pro398 and Leu399 from Z-site, Glu467 from γ-Glu site, and His461, part of the catalytic triad. Compound Lit_C208 shows potential selective inhibition against TvTIM over HsTIM, with favorable energy contributions toward TvTIM catalytic dyad, but away from HsTIM catalytic dyad. Compound Lit_C388 was most stable in FhCatL with a higher calculated binding energy by MMPBSA analysis than HsCatL, though not interacting with catalytic dyad, holding favorable energy contribution from residues oriented at FhCatL catalytic dyad. Therefore, these kinds of compounds are good candidates to continue researching and confirming their activity through in vitro studies as new selective antiparasitic agents. [ABSTRACT FROM AUTHOR]

Fascioliasis is a liver fluke disease in which food and water are the transmitting agents. The disease is caused by a genus of Fasciola, parasitic Trematoda. The genus Fasciola includes two species of Fasciola gigantica and Fasciola hepatica. The study of this disease is of particular importance as the infected cases are globally growing, especially in endemic areas. In this paper, a fractional‐order mathematical model is presented for fascioliasis disease. The optimization method is an effective choice in solving mathematical models of epidemic diseases. The study aims at presenting a new set of generalized Fibonacci polynomials as basic functions and some innovative operational matrices of derivatives in the Caputo sense to solve the fascioliasis disease model. The numerical experiments reveal better results and simulations show effectiveness of the presented method. [ABSTRACT FROM AUTHOR]

Background: Fascioliasis, caused by Fasciola hepatica, is a neglected zoonotic food-borne trematodiasis. The Caspian littoral in northern Iran is endemic for the disease, and human fascioliasis is well-known in that region. In the present study, we report the diagnosis, identification, and clinical management of a human case of fascioliasis associated with common bile duct (CBD) obstruction from a non-endemic remote area in southeastern Iran. Case presentation: A 42-year-old female was admitted to Afzalipour Medical Center hepatobiliary surgery ward in Kerman with abdominal pain for the past three months. Dilated biliary tract and an ill-defined mass in CBD were reported in abdominal ultrasonography and magnetic resonance cholangiopancreatography, respectively. During distal CBD operation, nine leaf-like motile flatworms were isolated. A morphological study confirmed all the isolates as Fasciola, and further molecular investigations, identified the flukes as F. hepatica using both pepck multiplex PCR and cox1 sequencing. Conclusion: Molecular and morphological findings of the study indicated the presence of human fascioliasis in the southeastern province of Sistan and Baluchestan in Iran. Fascioliasis is among the etiologies of chronic cholecystitis, and physicians should consider chronic cholecystitis associated with fascioliasis in the differential diagnosis. In the present report, endoscopic ultrasound was usefully applied for the accurate diagnosis of biliary fasciolosis. [ABSTRACT FROM AUTHOR]

Background. Fruits are a vital part of human nutrition because they contain the growth factors required to maintain normal health. Fruits are also known to harbour a wide range of parasites and bacteria. Consumption of unwashed raw fruits can lead to foodborne pathogens. This study was carried out to examine the presence of parasites and bacteria on fruits sold at two major markets in Iwo, Osun state, South-West Nigeria. Methods. Twelve different fresh fruits and seven different fresh fruits were purchased from different vendors from "Odo-ori" and "Adeeke" markets, respectively. The samples were transported to the microbiology laboratory of Bowen University, Iwo, Osun state for bacteriological and parasitological analysis. The parasites were concentrated by sedimentation and were examined using a light microscope, while for the microbial analysis, culturing and biochemical tests were also carried out on all the samples. Results. The parasites discovered include Fasciola hepatica eggs, Ascaris lumbricoides and Strongyloides stercoralis larvae, hookworm larvae, and Taenia and Trichuris trichiura eggs. Ascaris lumbricoides was most frequently detected (40.0%). Bacteria isolated from the sampled fruits include Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter aerogenes, Escherichia coli, Kluyvera ascorbata, Proteus mirabilis, Staphylococcus intermedius, Staphylococcus sp., Micrococcus sedentarius, Corynebacterium diphtheriae, and Streptococcus zooepidemicus. Conclusion. The presence of parasites and bacteria on the fruits observed indicates that public health diseases may arise from their consumption. Awareness and education among farmers, vendors, and consumers about the importance of personal and food hygiene through proper washing or disinfection of fruits could reduce the risk of parasites and bacterial fruit contamination. [ABSTRACT FROM AUTHOR]