The liver fluke Opisthorchis viverrini (Poirier, 1886) (Digenea) secretes extracellular vesicles (EVs) bearing CD63-like tetraspanins on their surface. Fluke EVs are actively internalised by host cholangiocytes in the bile ducts, where they drive pathology and promote neoplasia through induction of cellular proliferation and secretion of inflammatory cytokines. We investigated the effects of tetraspanins of the CD63 superfamily by co-culturing recombinant forms of the large extracellular loop (LEL) of O. viverrini tetraspanin-2 (rLEL-Ov-TSP-2) and tetraspanin-3 (rLEL-Ov-TSP-3) with non-cancerous human bile duct (H69) and cholangiocarcinoma (CCA, M213) cell lines. The results showed that cell lines co-cultured with excretory/secretory products from adult O. viverrini (Ov-ES) underwent significantly increased cell proliferation at 48 hours but not 24 hours compared to untreated control cells (P < 0.05), whereas rLEL-Ov-TSP-3 co-culture resulted in significantly increased cell proliferation at both 24 hours (P < 0.05) and 48 hours (P < 0.01) time points. In like fashion, H69 cholangiocytes co-cultured with both Ov-ES and rLEL-Ov-TSP-3 underwent significantly elevated Il-6 and Il-8 gene expression for at least one of the time points assessed. Finally, both rLEL-Ov-TSP-2 and rLEL-Ov-TSP-3 significantly enhanced migration of both M213 and H69 cell lines. These findings indicated that O. viverrini CD63 family tetraspanins can promote a cancerous microenvironment by enhancing innate immune responses and migration of biliary epithelial cells.
A total of 1,724 beef and 2,941 dairy cattle older than one year from 66 beef and 67 dairy farms in the Czech Republic were examined for the presence of rumen and liver fluke eggs in 2019-2022. Out of 227 positive animals, all were positive for paramphistome and five for fasciolid eggs. Molecular analysis of the ITS2 rDNA revealed the presence of Calicophoron daubneyi (Dinnik, 1962) and Fasciola hepatica Linnaeus, 1758. Faecal egg count (FEC) showed low infection intensity (12 EPG) in animals infected with F. hepatica and high variability in C. daubneyi infections (2-589 EPG). Efficacy of oxyclozanide, albendazole, ivermectin, and closantel against C. daubneyi infection was evaluated at eight beef cattle herds. Faecal samples were collected from all positive animals at 0 and 21days post-treatment. Based on FEC, albendazole, ivermectin and closantel reduced the number of C. daubneyi eggs shed by 0-9.9%, with no effect on the number of infected animals. The use of oxyclozanide on two beef farms showed 100% efficacy against C. daubneyi and F. hepatica. Follow-up examination 5-6 months after drug application showed reinfection of most animals with C. daubneyi, but the FEC was significantly lower. The finding of four dairy cows infected with C. daubneyi housed in a stable without pasture suggests the possibility of the infection being introduced through roughage.
- MeSH
- Albendazole MeSH
- Fasciola hepatica * genetics MeSH
- Feces MeSH
- Trematode Infections * epidemiology veterinary MeSH
- Ivermectin therapeutic use MeSH
- Oxyclozanide MeSH
- Paramphistomatidae * genetics MeSH
- Prevalence MeSH
- Cattle MeSH
- Trematoda * MeSH
- Animals MeSH
- Check Tag
- Cattle MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
Introduction: Livestock, including sheep, are infected with many types of parasitic diseases, and among these diseases is fascioliasis, or the so-called liver rot, which is caused by the Fasciola hepatica worm. This disease is widespread in many regions of the world, especially those with moderate climatic conditions. This disease harms the state's economy and livestock breeders due to the high rates of infection, which greatly affects animal production.Materials: Wax sections were made from F. hepatica, cut with a thickness of 7 microns, and stained with Hematoxylin and Eosin stain and Von Kossa technique to investigate the calcium sites in different organs of the worm.Results: There was a clear discrepancy in the density of calcium present in several internal organs of F. hepatica.Conclusion: We infer from this research that there is a clear variation in the density of calcium as well as the size and shape of the calcareous corpuscles in their different locations in the worm's organs.
Fasciolóza je parazitární onemocnění způsobované dvěma druhy motolic rodu Fasciola. Jedná se o jednu z nejvýznamnějších parazitóz hospodářských zvířat, vzácná není ani nákaza člověka. Masivní používání léčiva triklabendazolu vedlo k tvorbě rezistentních kmenů fasciol, které jsou v současnosti velmi rozšířené. Proto se věnuje mimořádné úsilí vývoji vakcíny. Proteolytický systém fasciol se účast-ní řady procesů nezbytných pro parazita, od líhnutí miracidií přes pronikání tělem hostitele a ovlivňování jeho fyziologických procesů až po trávení živin. Z tohoto důvodu jsou proteolytické enzymy atraktivním cílem pro vývoj vakcín proti fasciolóze. Tento článek pojednává o nejdůležitějších proteasách fasciol a jejich použití jako vakcinačních antigenů.
Fasciolosis is a parasitic infection caused by two species of flukes from the genus Fasciola. It is considered one of the most important diseases of livestock, but human infection is also common. Massive usage of the drug triclabendazole led to the development of resistant flukes, which are currently spread out in many countries. Hence there is much effort put into development of vaccines against fasciolosis. Proteolytic system of Fasciola plays crucial role in number of processes such as hatching, tissue invasion, nutrient intake or modulation of the host physiology. For that reason proteases of Fasciola are attractive targets for vaccine development. This paper summarizes current knowledge about the most important proteases from Fasciola hepatica and their vaccination potential.
- MeSH
- Fasciola hepatica * enzymology parasitology growth & development MeSH
- Fascioliasis * physiopathology prevention & control MeSH
- Animals, Domestic MeSH
- Cathepsin B antagonists & inhibitors chemistry therapeutic use MeSH
- Cathepsin L chemistry immunology therapeutic use MeSH
- Vaccine Development MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
BACKGROUND: Representatives of the trematode family Fasciolidae are responsible for major socio-economic losses worldwide. Fascioloides magna is an important pathogenic liver fluke of wild and domestic ungulates. To date, only a limited number of studies concerning the molecular biology of F. magna exist. Therefore, the objective of the present study was to determine the complete mitochondrial (mt) genome sequence of F. magna, and assess the phylogenetic relationships of this fluke with other trematodes based on the mtDNA dataset. FINDINGS: The complete F. magna mt genome sequence is 14,047 bp. The gene content and arrangement of the F. magna mt genome is similar to those of Fasciola spp., except that trnE is located between trnG and the only non-coding region in F. magna mt genome. Phylogenetic relationships of F. magna with selected trematodes using Bayesian inference (BI) was reconstructed based on the concatenated amino acid sequences for 12 protein-coding genes, which confirmed that the genus Fascioloides is closely related to the genus Fasciola; the intergeneric differences of amino acid composition between the genera Fascioloides and Fasciola ranged 17.97-18.24 %. CONCLUSIONS: The determination of F. magna mt genome sequence provides a valuable resource for further investigations of the phylogeny of the family Fasciolidae and other trematodes, and represents a useful platform for designing appropriate molecular markers.
- MeSH
- Fasciola hepatica chemistry classification genetics isolation & purification MeSH
- Fasciolidae chemistry classification genetics isolation & purification MeSH
- Phylogeny MeSH
- Genome, Mitochondrial * MeSH
- Genome, Helminth * MeSH
- Trematode Infections parasitology MeSH
- Molecular Sequence Data MeSH
- Helminth Proteins chemistry genetics MeSH
- Base Sequence MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
1. Giant liver fluke Fascioloides magna is a dangerous parasite, which infects herbivores. It was imported to Europe from North America and started to spread. Benzimidazoles like albendazole, mebendazole, triclabendazole and salicylanilides closantel and rafoxanide are the most used anthelmintics to control fascioloidosis. However their effect might be altered via drug-metabolizing enzymes of this parasite. 2. The aim of our study was to determine the activities of drug-metabolizing enzymes in F. magna and the metabolism of above mentioned anthelmintics. 3. Activities of several oxidative, reductive and conjugative enzymes towards various model xenobiotic substrates were found in F. magna subcellular fractions. 4. Subcellular fractions from F. magna oxidized albendazole to its sulphoxide metabolite and reduced mebendazole to hydroxyl-mebendazole. Under ex vivo conditions, only very-low concentrations of these compounds were detected using high-performance liquid chromatography/mass spectrometry. 5. The results indicate that the giant liver fluke possesses the active xenobiotic-metabolizing system. The overexpression of this system may play an important role in parasite resistance against these anthelmintics.
- MeSH
- Albendazole metabolism MeSH
- Anthelmintics metabolism MeSH
- Benzimidazoles metabolism MeSH
- Fasciola hepatica drug effects enzymology MeSH
- Mebendazole metabolism MeSH
- Rafoxanide metabolism MeSH
- Salicylanilides metabolism MeSH
- Sulfoxides metabolism MeSH
- Chromatography, High Pressure Liquid MeSH
- Xenobiotics metabolism MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
Infection with Fascioloides magna (Digenea) causes serious damage to liver tissue in definitive hosts represented by ruminants, especially cervids. The distribution of F. magna includes the indigenous areas in North America, and the areas to which F. magna was introduced-Central Europe, Southeast Europe, and Italy. The North American intermediate host of F. magna, the freshwater snail Pseudosuccinea columella (Lymnaeidae), is an invasive species recorded in South America, the Caribbean, Africa, Australia, and west and Southeast Europe. In Europe, Galba truncatula is the snail serving for transmission, but P. columella has potential to become here a new intermediate host of F. magna. Little is known about interactions between F. magna and P. columella. In this study, the susceptibility of P. columella (Oregon, USA) to the infection by a single miracidium of the Czech strain of F. magna and the influence of F. magna on snail fecundity, shell height, and survival were evaluated. The data show that the Oregon strain of P. columella is a highly suitable host for the Czech strain of F. magna, with the infection rate of 74 %. In addition, a negative effect on survival rate of infected snails was recorded only in the late phase of infection. The infection was accompanied by a major reduction in egg mass production and by a decrease in the number of eggs per egg mass. The shell height of infected snails did not significantly differ from that in unexposed controls.
- MeSH
- Species Specificity MeSH
- Fasciola hepatica classification genetics pathogenicity MeSH
- Fascioliasis epidemiology parasitology veterinary MeSH
- Fertility MeSH
- Snails parasitology MeSH
- Liver parasitology pathology MeSH
- Survival Rate MeSH
- Molecular Typing MeSH
- Ovum parasitology MeSH
- Deer parasitology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Africa epidemiology MeSH
- Australia epidemiology MeSH
- Europe epidemiology MeSH
- South America epidemiology MeSH
- Caribbean Region epidemiology MeSH
- North America epidemiology MeSH
We have studied the influence of both levamisole (AL) and Freund's adjuvant (AF) on the immunisation of mice with the secretory antigens of adults of the liver fluke Fasciola hepatica Linnaeus, 1758. Total IgG antibodies were detected in all groups where the F. hepatica antigen was administered, been levels of IgG1 increased respect to IgG2a antibodies. During immunisation, IL-4 and IFN-γ were only detected in AL and AF groups, but after infection, IL-4 boosted in all groups. IFN-γ increased two fold in AF and AL groups compared to the saline solution (AS) group. Worm recovering was of 32-35% in groups administered without antigen whereas in AS, AL and AF groups recovering was of 25%, 12% and 8%, respectively. Macroscopical lesions in the liver were scarce in AL and AF groups. Our data suggest that immunisation of mice with antigens of F. hepatica enhances the immune response avoiding both liver damage and worm establishment after challenge infection. The murine model of fasciolosis has appeared to be useful to elucidate the mechanism by which the parasite modulates immune responses toward a Th2 type but also the development of Th1 type-inducing vaccines.
- MeSH
- Antigens, Helminth immunology blood MeSH
- Fasciola hepatica immunology pathogenicity MeSH
- Fascioliasis drug therapy immunology MeSH
- Freund's Adjuvant immunology MeSH
- Immunomodulation drug effects MeSH
- Liver pathology drug effects MeSH
- Kinetics MeSH
- Levamisole * immunology MeSH
- Lymnaea parasitology MeSH
- Metacercariae MeSH
- Models, Animal MeSH
- Mice MeSH
- Parasite Load MeSH
- Statistics as Topic MeSH
- In Vitro Techniques methods MeSH
- Body Weight MeSH
- Organ Size MeSH
- Check Tag
- Mice MeSH
- Publication type
- Journal Article MeSH
Experimental infections of Galba sp. (origin, Colombia) with allopatric isolates of Fasciola hepatica from France or Fascioloides magna from the Czech Republic were carried out during five successive snail generations to determine if this lymnaeid might sustain complete larval development of either parasite. In snails exposed to F. hepatica, 7 of 400 snails harboured several rediae and only two snails contained a small number of free cercariae on day 50 post-exposure. In contrast, the intensity of F. magna infection in Galba sp. progressively increased from the F1 to F5 generations. Spontaneous cercarial shedding of F. magna occurred in 7 of 100 Galba sp. belonging to the F5 generation and the number of shed cercariae did not differ significantly from that noted in control Galba truncatula of French origin. Galba sp. from Colombia can be added to the list of potential intermediate hosts for F. magna.
- MeSH
- Fasciola hepatica growth & development physiology MeSH
- Fasciolidae growth & development physiology MeSH
- Fascioliasis parasitology veterinary MeSH
- Snails classification growth & development parasitology MeSH
- Disease Reservoirs parasitology MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Geographicals
- Czech Republic MeSH
- France MeSH
BACKGROUND: Population structure and genetic interrelationships of giant liver fluke Fascioloides magna from all enzootic North American regions were revealed in close relation with geographical distribution of its obligate definitive cervid hosts for the first time. METHODS: Variable fragments of the mitochondrial cytochrome c oxidase subunit I (cox1; 384 bp) and nicotinamide dehydrogenase subunit I (nad1; 405 bp) were applied as a tool. The concatenated data set of both cox1 and nad1 sequences (789 bp) contained 222 sequences that resulted in 50 haplotypes. Genetic data were analysed using Bayesian Inference (BI), Maximum Likelihood (ML) and Analysis of Molecular Variance (AMOVA). RESULTS: Phylogenetic analysis revealed two major clades of F. magna, which separated the parasite into western and eastern populations. Western populations included samples from Rocky Mountain trench (Alberta) and northern Pacific coast (British Columbia and Oregon), whereas, the eastern populations were represented by individuals from the Great Lakes region (Minnesota), Gulf coast, lower Mississippi, and southern Atlantic seaboard region (Mississippi, Louisiana, South Carolina, Georgia, Florida) and northern Quebec and Labrador. Haplotype network and results of AMOVA analysis confirmed explicit genetic separation of western and eastern populations of the parasite that suggests long term historical isolation of F. magna populations. CONCLUSION: The genetic makeup of the parasite's populations correlates with data on historical distribution of its hosts. Based on the mitochondrial data there are no signs of host specificity of F. magna adults towards any definitive host species; the detected haplotypes of giant liver fluke are shared amongst several host species in adjacent populations.
- MeSH
- Fasciola hepatica classification enzymology genetics isolation & purification MeSH
- Fascioliasis epidemiology parasitology veterinary MeSH
- Phylogeny MeSH
- Genetic Variation MeSH
- Molecular Sequence Data MeSH
- Ruminants parasitology MeSH
- Helminth Proteins genetics metabolism MeSH
- Electron Transport Complex IV genetics metabolism MeSH
- Deer MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Quebec MeSH
- United States MeSH
