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PubMed2000

Investigation of Fasciola hepatica sample preparation for two-dimensional electrophoresis.

This paper investigates the preparation of Fasciola hepatica samples for two-dimensional electrophoresis (2-DE). Whole samples were prepared by both hot sodium dodecyl sulfate (SDS) solubilisation and precipitation using trichloroacetic acid (TCA) to remove nonprotein contaminants and to inactivate endogenous proteases. Sample preparation had a marked influence on the 2-DE gel profile. TCA precipitation resulted in no measurable improvement in the profile observed, compared to the untreated control. Solubilisation of sample with hot SDS increased the number of protein spots, as did TCA precipitation with the addition of phosphotungstic acid. The preparation of excretory-secretory (ES) products poses problems due to both high salt concentrations and low protein concentration. All precipitation methods used to overcome this gave similar profiles, except acetone alone, which caused depletion of the larger proteins. TCA in acetone gave the best result, similar to that obtained by centrifugal filtration of the sample. Overcrowding of spots in some regions of the 2-DE gel occurred in the whole Fasciola hepatica sample. This problem was alleviated by differential solubilisation, which also resulted in the enrichment of some proteins.

Animals↗
PubMed2000

Obstruction of common bile duct caused by liver fluke--Fasciola hepatica.

Three cases of obstruction of the common bile duct by Fasciola hepatica with two of the patients presenting jaundice are reported. The authors have reviewed several publications concerning common bile duct obstruction by liver fluke, a quite rare complication of fascioliasis. Only nineteen cases of common bile duct obstruction caused by Fasciola hepatica have been reported in a review of medical publications during last ten years. Clinical presentation, diagnostic methods and considerations, types of surgery are fairly uniform in all of the reported cases. Almost all of patients reviewed, had the history, symptoms and signs characteristic for cholelithiasis including recurrent colic pain in right hypochondriac area, fever or subfebrile temperature, fluctuating or stabile jaundice, and palpable painful gallbladder. The laboratory findings in all cases reviewed had shown leucocytosis, eosinophilia, high or slight elevated serum bilirubin. Echographically commonly revealed dilated intra- and extrahepatic bile ducts containing one or more hyperechogenic elements with or without casting an acoustic shadow. All patients underwent open surgery, comprised with choledochotomy and if possible extraction of the fluke. Only two postoperative cases were of necessity followed by ERCP. In all of our cases the primary pre-operative diagnosis was choledocholithiasis, with diagnose of fascioliasis established at the operation. According to the literature this uncertainty in diagnosis is common because of difficulties in differentiation of fascioliasis versus choledocholithiasis. Considerations for making the differential diagnosis--a history of origin or visiting in endemic area of infection, history of eating of aquatic vegetables, laboratory findings including eosinophilia, fasciola eggs in stool, sonography and radiological imaging results and enzyme-linked immunosorbent essay (ELISA) which has been shown to be rapid, sensitive and quantitative. In all three cases we have observed intraoperative significant signs for liver fascioliasis to include surface scarring of the left lobe on the liver--tracks caused by subcapsular migration and location of the hepatic lesions (these findings were also seen by two authors in literature) with resemblance to Japanese letters. The most effective drug for treatment of fascioliasis according to our experience and literature reviewed is bithionol.

Adult↗
PubMed1992

Comparison of an indirect haemagglutination assay and an ELISA for diagnosing Fasciola hepatica in experimentally and naturally infected sheep.

An enzyme-linked immunosorbent assay (ELISA) with somatic (S) or excretory-secretory antigens (ES) was compared with an indirect haemagglutination assay (IHA) for ability to detect antibodies against Fasciola hepatica in sheep. The specificity of both assays was determined by testing sera collected from sheep experimentally or naturally mono-infected with Fasciola hepatica, Haemonchus contortus, Ostertagia circumcincta, Cooperia curticei, Taenia ovis, Eimeria spp., Trichostrongylus vitrinus, Trichostrongylus colubriformis or Nematodirus battus respectively. With S or ES antigens the specificity of the ELISA was 98% or 95% respectively, whereas the specificity of the IHA was 86%. Antibodies directed against Fasciola hepatica were detected by the ELISA with S or ES antigens from 2 weeks after infection until the end of the experiment, whereas the IHA detected antibodies from week 3. We conclude that the ELISA with S antigens compares favourably with the IHA and can be used for the serodiagnosis of ovine fasciolosis in the Netherlands.

Animals↗
PubMed1987

Studies on heterologous resistance between Schistosoma mansoni and Fasciola hepatica in inbred rats.

Fischer rats infected with Fasciola hepatica showed significant resistance to Schistosoma mansoni challenge, and vice versa, whereas immunization with 20 Krad-irradiated S. mansoni cercariae failed to protect against F. hepatica challenge, but did protect against homologous challenge. When groups of rats received intraperitoneal implants of newly excysted juvenile flukes, 20- to 22-day-old juveniles, or 8- to 10-week-old flukes, none was significantly protected against S. mansoni challenge, whereas juvenile implants did protect against homologous F. hepatica challenge. In passive transfer experiments in rats, serum from F. hepatica-infected rats or rabbits protected recipients against homologous, but not heterologous challenge, and serum from rats vaccinated with 20 Krad-irradiated S. mansoni cercariae protected recipients against homologous, but not heterologous challenge. These experiments provide evidence that the mechanisms involved in homologous and heterologous resistance are different, the latter lacking immunological specificity. Microsphere injections in F. hepatica-infected rats demonstrated 'shunting' from the portal system to the systemic circulation. If migrating schistosomula are also 'shunted' in Fasciola-infected rats, this, rather than immunologically specific effector mechanisms, might explain their failure to establish in the portal system.

Animals↗
PubMed2006

Fasciola hepatica infection: clinical and computerized tomographic findings of ten patients.

BACKGROUND/AIMS: Fasciola hepatica is the cause of liver infection, fascioliasis. Although rare, it is still a problem even in developed countries. In this study, the clinical and computerized tomographic findings of 10 patients diagnosed with fascioliasis are summarized. METHODS: The medical records of the patients with fascioliasis were retrospectively examined. Clinical, laboratory findings and computerized tomographic results were recorded. RESULTS: Abdominal pain, fever, eosinophilia and abnormal liver function tests were the most commonly encountered symptoms and signs. One patient was human immunodeficiency virus -positive with active tuberculosis. Serologic test for fasciola hepatica was positive in all patients. Nodular masses without prominent enhancement, and branching low-attenuated tubular lesions were the most commonly seen tomographic findings and were supportive for the diagnosis. All except the HIV-positive patient received bithionol therapy; six patients responded well, two lost contact with the clinic and one patient who was unresponsive to bithionol therapy received triclabendazole. During follow-up of the six patients who responded, all the clinical and radiological findings regressed. CONCLUSION: In any patient with peripheral eosinophilia, abdominal pain and elevated liver enzymes, especially when CT reveals tubular and nodular hypodense lesions particularly in subcapsular area, F. hepatica infection should be considered. Either triclabendazole or bithionol can be used effectively for the treatment.

Abdominal Pain↗
PubMed1995

Detection of excretory-secretory circulating antigens in sheep infected with Fasciola hepatica and with Schistosoma mansoni and F. hepatica.

Two groups of sheep were exposed to Fasciola hepatica and Schistosoma mansoni/F. hepatica infections, respectively, to determine the time of appearance of F. hepatica circulating excretory-secretory (FhES) antigens. Five sheep were infected orally with 400 F. hepatica metacercariae. Five additional sheep were infected first percutaneously with 5000 S. mansoni cercariae, and 10 weeks later orally with 400 F. hepatica metacercariae. Antigen detection was performed in the two groups by a double antibody microELISA. In the group infected with F. hepatica, circulating FhES antigens were detected in all five animals. In the group infected first with S. mansoni none of the sera had positive optical density values for FhES antigens throughout the 10 week period following infection. After F. hepatica challenge, FhES antigens were detected in all animals. Thus, sheep infected with S. mansoni did not develop detectable F. hepatica circulating antigens during the 10 week period following Schistosoma infection. Anti-FhES antibodies were detected by FAST-ELISA, and all animals infected with F. hepatica were positive. In the group infected with S. mansoni, cross-reactive antibodies to F. hepatica were detected by 4-6 weeks post infection. All sheep with double infections showed high antibody levels by 2 weeks after infection with F. hepatica.

Animals↗
PubMed2002

[Infection with fasciola hepatica causing elevated liver-enzyme results and eosinophilia - serologic and endoscopic diagnosis and therapy].

A 36 year old man was diagnosed with eosinophilia and elevated liver-enzyme levels in a preoperative routine laboratory testing. In the course of the evaluation we found high levels of Fasciola hepatica specific antibodies which lead to an ERC with biliary aspiration after choleretic stimulation. The bile showed Fasciola eggs and subsequently we recovered a parasite from the bile duct. Abdominal CT- and MRI-Scans showed lesions of the liver consistent with hepatic fascioliasis. Focussed patient's history revealed ingestion of contaminated watercress in Turkey as the most likely source of infection. For patients with elevated liver-enzymes of unknown etiology with eosinophilia or in combination with a congruent patient's history even without eosinophilia serologic studies of Fasciola hepatica seem to be advisable. We suggest an ERC with biliary aspiration to prove the diagnosis in case of positive results, if the prepatency period of 4 months after the ingestion is over. Fasciola hepatica is rarely diagnosed in Germany probably due to the lack of awareness of the disease.

Adult↗
PubMed1957

The effect of lysergic acid diethylamide, 5-hydroxytryptamine, and related compounds on the liver fluke, Fasciola hepatica.

The rhythmical activity of the liver fluke, Fasciola hepatica, was stimulated by 5-hydroxytryptamine and by lysergic acid diethylamide at very low concentrations. The effect was peripheral and was not mediated through the central ganglion. Other amines also stimulated rhythmical activity, the most potent being the indolamines.Bromolysergic acid diethylamide, and other analogues such as yohimbine, harmine, and dopamine depressed rhythmical movement and antagonized the stimulant action of 5-hydroxytryptamine and lysergic acid diethylamide. Evidence which suggests the presence of tryptamine receptors in the trematode is discussed.

Animals↗
PubMed2002

[The influence of Fasciola hepatica invasion on the assorted milk quality parameters in cows].

The investigations of the influence of Fasciola hepatica invasion on the assorted parameters of milk quality were performed in cows. Mean protein content in the milk from infected animals ammounted 3.60% and was higher than in the milk from uninfected cows (3.30%). Similar tendency was observed in the case of the fat (5.08% and 3.91%) and lactose (4.36% and 4.34%). The count of somatic cells in milk from infected cows was lower than in milk from uninfected animals (356.07 and 1776.92). It can indicate immunosupressive activity of Fasciola hepatica on the host.

Animals↗
PubMed1981

Prevalence of Fasciola hepatica infection in native mammals in southeastern Australia.

Fasciola hepatica infection was found in 5 of 12 marsupial and 1 of 3 eutherian species examined in southeastern Australia. Prevalence of infection in native mammals was as follows: Macropus giganteus, 59%; M. rufogriseus banksianus, 15%; Wallabia bicolor, 5%; Vombatus ursinus, 30%; Trichosurus vulpecula, 5%; and Rattus fuscipes, 4%. Collection sites were assigned to one of two categories. Prevalence was higher in macropodids collected on agricultural pastures grazed by sheep and cattle (category A) than in macropodid collected in forested areas variably grazed by livestock (category B). A high prevalence of infection (82%) was recorded in M. giganteus in an area from which sheep and cattle had been excluded for 15 years. Prevalence of F. hepatica infection in V. ursinus was similar in the two habitat categories; however, within forested areas infection was more common in V. ursinus collected near swamps and creeks than those taken from drier ridges. The presence of habitat suitable for the snail intermediate host. Lymnaea tomentosa, and utilization of this habitat by domestic and native mammals are important epidemiological factors. Moderate numbers of flukes (1-95) and faecal egg production (1-195 eggs per gram of faeces (EPG)) in M. giganteus and M.r. banksianus emphasize their suitability as hosts and contaminators of pasture. Small numbers of flukes and conspicuous hepatic lesions in V. ursinus suggest moderate resistance to infection. The low prevalence and small number of flukes in T. vulpecula was attributed to the arboreal habit of this host. R. fuscipes is susceptible to infection but infected individuals were found in only one of numerous forest areas with suitable snail habitat. Feeding behaviour is probably responsible for the absence of F. hepatica infection in W. bicolor and Rattus lutreolus in forested areas.

Animals↗
PubMed1997

A focus of Fasciola hepatica in Crete without human cases.

In Greece, Fasciola hepatica, the sheep liver fluke, is common in sheep but only three human cases of fascioliasis have been reported. An epidemiological study was conducted in central Crete which proved to be a focus of F. hepatica. Sheep and snails were found positive for this parasite, but none of the 205 persons tested serologically.

Animals↗
PubMed2003

Natural infections of Omphiscola glabra (Lymnaeidae) with Fasciola hepatica in central France.

As larval forms of Fasciola hepatica have periodically been detected in Omphiscola glabraafter their collection from watercress beds or from meadows since 1995, field investigations in 37 populations of O. glabra were carried from 1996 to 2002. This was done in order to determine the changes in prevalences and intensities of these natural infections with F. hepatica in relation to the type of snail habitat and the year of snail collection. Snails infected with F. hepatica were found in all samples made in swampy meadows and roadside ditches in all years. In fenced pools and walled gardens, snail infections were only found from 1998 and 1999 onwards, respectively. In the four types of habitats, the prevalences of F. hepatica infections increased slightly over time (0.8-2.1% for snails sampled in swampy meadows, for example) but this increase varied with the habitat. The mean shell heights of infected snails (6.2-7.8 mm) were similar whatever the type of habitat. The numbers of cercariae-containing rediae counted in snails sampled in swampy meadows, roadside ditches, and fenced pools significantly increased over time. Significant numerical variation between these redial burdens was also observed in relation to snail habitat. As the larval development of F. hepatica is facilitated by the presence of another trematode larval form ( Paramphistomum daubneyi), the finding of some naturally infected O. glabra in watering places known to have no contact with domestic or wild large mammals might be explained by the development of P. daubneyi in small mammals such as lagomorphs. However, a progressive adaptation of F. hepatica miracidia to O. glabra over time, which would permit the infection of snails at sizes larger than 2 mm, could not be excluded.

Animals↗
PubMed1996

Modification of accessory activity of peritoneal cells from Fasciola hepatica infected rats.

The effect of Fasciola hepatica excretory-secretory antigen (ESA) was studied in the modulation of the accessory functions of peritoneal cells (PC) of rats infected with the parasite. PC rats infected with F. hepatica 7 and 14 days previously showed a marked decrease in phagocytic activity against Candida tropicalis (P < 0.007 and P < 0.004, respectively). The same effect was observed when the assay was carried out with PC from animals injected 7 days before with ESA (P < 0.001) including PC previously treated in vitro with ESA. To investigate the effect of ESA on the antigen presenting ability, PC of animals infected before transfer to syngeneic normal rats were stimulated in vitro with either F. hepatica whole antigen (FhWA), ESA or human serum albumin (HSA). Delayed type hypersensitivity (DTH) response to the different antigens studied over a 35 day period was negative in rats transferred with sensitised PC. When these animals were immunised with the corresponding antigen the DTH response became positive. A similar result was obtained in PC receptors from ESA inoculated rats and in vitro stimulated with FhWA or HSA. These data suggest that the alterations observed in the functions of peritoneal cells may, in part be due to the effect of the F. hepatica ESA.

Animals↗
PubMed1991

[Intestinal cellular reaction of cattle after infection by Fasciola hepatica].

Defence reactions of cattle against Fasciola hepatica take place not only in the liver but also on a prehepatic level. The aim of the present experiments is to identify and quantify the cellular response in the small intestine. Five groups of two bulls (Simmental x Red Holstein, 6 months old) were infected by oesophageal probe with metacercariae of F. hepatica (MC) according to two experimental procedures. Two groups of bulls served as controls. In the first experiment both infected groups received 1000 MC once or twice at an interval of five weeks. The intestinal tissue was taken before infection and 5, 10 and 19 weeks p.i., respectively 5 and 14 weeks p. reinf. (p.ri.). In the second experiment, three infected groups received 400, 2 x 400 MC (at an interval of five weeks) and 800 MC respectively. Biopsies were taken one week ante infectionem, 6, 11 and 19 weeks p.i., respectively 1, 6 and 14 weeks post reinfectionem (p. ri.). The biopsies of the small intestine were prepared for histology and the tissue sections stained as follows: Astra blue for mucosal mast cells (MMC), Lendrum for eosinophils (EOS) and Periodic Acid Schiff (PAS) for goblet cells (GC). The density of the three cell populations in the mucosa was determined microscopically and quantified. In both experiments the density of the MMC and EOS increased with each reinfection and depended on the infectious dose. There was no significant variation of the GC-numbers. The described intestinal cellular reactions suggest a participation of the small intestinal wall in the immunity against the common liver fluke.

Animals↗
PubMed2001

Early immunodiagnosis of fasciolosis in ruminants using recombinant Fasciola hepatica cathepsin L-like protease.

A diagnostic ELISA with recombinant Fasciola hepatica cathepsin L-like protease as antigen was developed to detect antibodies against F. hepatica in sheep and cattle. The recombinant cathepsin L-like protease was generated by functional expression of the cDNA from adult stage F. hepatica flukes in Saccharomyces cerevisiae. Specificity and sensitivity of the cathepsin L enzyme-linked immunosorbent assay (ELISA) was assessed using sera from sheep and calves experimentally or naturally mono-infected with F. hepatica and six-seven other parasites. The sensitivity of the cathepsin L ELISA for sheep and cattle sera was 99.1 and 100%, respectively. In the experimental setting with established mono-infections, the specificity of the cathepsin L ELISA was 98.5% for cattle sera and 96.5% for sheep sera. In experimentally infected cattle and sheep, the first detection of F. hepatica-specific antibodies appeared first between 5 and 7 weeks post-infection, but depended on the infectious dose of F. hepatica. In ELISA the detection preceded first detection of the infection based on egg counts and remained detectable till at least 23 weeks after a primary F. hepatica infection. Detection of Fasciola gigantica infections was similar to detection of F. hepatica. The first detection occurred at week 5 and signals persisted for at least 20 weeks. All sera from naturally F. hepatica infected sheep were seropositive in the cathepsin L-like ELISA. The relevance of this ELISA format was also evaluated using sera from naturally infected cattle in the Netherlands, Ecuador and Vietnam and compared with results from egg-counts. For the latter two endemic areas with mixed parasitic infections the 'apparent' sensitivity of the cathepsin L ELISA was calculated for all serum samples together to be 90.2%. The 'apparent' specificity under these conditions was calculated to be 75.3%. In cattle, the cathepsin L ELISA was superior to the concurrently evaluated peptide ELISA format using a single epitope as the antigen both in controlled natural infections as well as in infections in endemic areas. The present ELISA-format contributes a relatively sensitive and reliable tool for the early serodiagnosis of bovine and ovine fasciolosis.

Animals↗
PubMed1984

Early migration of immature Fasciola hepatica and associated liver pathology in cattle.

Immature Fasciola hepatica were successfully recovered from the body cavity of cattle from two to 21 days after oral infection. Flukes had penetrated the liver in large numbers by day 7 but there did not appear to be a corresponding reduction in the numbers recovered from the body cavity. Liver pathology showed a typical progressive tissue disruption with a neutrophil and lymphocyte infiltration by day 7 and a marked eosinophil response from day 14. The mesothelium of the liver capsule underwent metaplasia, becoming ultimately highly active and columnar.

Animals↗
PubMed1976

A technique for in vivo labelling of Fasciola hepatica miracidia with radioselenium.

Radioactivity is incorporated in Fasciola hepatica and its eggs when 75Semethionine is administered intraperitoneally in chronically infected mice. Maximum egg-bound radioactivity is achieved 2-3 days after the administration of the labelled aminoacid. Seventy-three per cent of the radioactivity of the egg is recovered in the free-swimming miracidium. There appears to be no altered motility or infectivity to snails of miracidia made radioactive at a level corresponding to 6 pCi per larva. Suggestions are made for the application of labelled larvae in studies on various aspects of the parasite-snail contact.

Animals↗