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PubMed1977

A study of a strategic dosing programme against ovine fascioliasis on a hill farm.

In a farm-scale veterinary preventive programme run from 1973 to 1976 the use of rafoxanide (Flukanide; Merck Sharp & Dohme) in the prophylaxis of ovine fascioliasis resulted in a marked reduction in the percentage of ewes passing fluke eggs in the faeces and an absence of clinical cases of fascioliasis throughout these years. Three separate flocks of hill sheep were used in this study in which the aim of the dosing programme followed was to kill the majority of liver flukes (Fasciola hepatica) before they reached adult egg laying stage, thereby eliminating, or considerably reducing, the number of eggs passed on to the pasture.

Animals↗
PubMedDate not supplied

Hepatic fascioliasis: case report and review.

A well documented case of hepatic fascioliasis (HF), successfully treated with triclabendazole, is reported. Predominant clinical manifestations were fever, marked eosinophilia and abdominal pain. Triclabendazole was given as two single oral doses of 10 mg/kg each. Neither side effects nor clinical or parasitological relapses were seen after three months of follow up. Based on this experience and few other similar reports in the literature, triclabendazole might be a valid therapeutical alternative in the treatment of human fascioliasis.

Administration, Oral↗
PubMed2000

[Fecal antigens of Fasciola hepatica potentially useful in the diagnosis of fascioliasis].

This study describes which antigens of Fasciola hepatica are present in the feces of patients with chronic fascioliasis and in the feces of rats infected experimentally with F. hepatica metacercariae. Using a Western blot assay technique with hyperimmune serum obtained from excretory-secretory antigens of adult F. hepatica, we found in the patients' feces antigens of possible diagnostic interest, with molecular weights of 14, 19, 20, 23, 25, 32, 46, 51, and 62 kilodaltons (kDa). In addition, we showed that the peptides of 14, 20, 23, and 51 kDa are also recognized by the majority of the sera from chronic patients. We used affinity chromatography to purify the antigens present in the feces of rats that had been infected for 6 to 12 weeks, using ES78 monoclonal antibody bound to CNBr-activated Sepharose 4B. Through that approach, we identified six polypeptides, of 11, 14, 26, 32, 47, and 51 kDa; three more polypeptides, of 17, 24, and 66 kDa, could only be identified in the feces of rats that had been infected for 10 to 12 weeks. Our results suggest that these polypeptides could be antigens common to both parasitic stages. This is particularly true for the polypeptides of 14, 24, 26, and 51 kDa, because they reacted with the immune sera, the human sera, and the ES78 monoclonal antibody. These polypeptides could be important markers for acute and chronic fascioliasis.

Animals↗
PubMed2003

Optimized serodiagnosis of sheep fascioliasis by Fast-D protein liquid chromatography fractionation of Fasciola hepatica excretory-secretory antigens.

Current methods for the serodiagnosis of sheep fascioliasis show suboptimal sensitivity, specificity, or both. With the aim of developing an improved method, we fractionated native Fasciola hepatica excretory-secretory antigens (ESAs) by size-exclusion FPLC (fast protein liquid chromatography) on a Superdex 75 HR 10/30 column and then tested the serodiagnostic value of the antigens contained in each one of the 4 peaks obtained (peaks I-IV). Serodiagnostic value was assessed using sera from sheep naturally infected with F. hepatica (group A); sera from the individuals of a fluke-free herd (most of which also had other intestinal nematodes, lung nematodes, Moniezia spp., and/or Cysticercus tenuicollis) sera from a fluke-free herd (group B); sera from lambs experimentally infected with 10-40 F. hepatica metacercariae (group C); and sera from uninfected control lambs (group D). Enzyme-linked immunosorbent assay (ELISA) with peak I or II as target antigens (and to a lesser extent with peak III as target) showed reactivity with negative sera, so that it was not possible to establish cutoff values discriminating infected and uninfected animals. In contrast, when peak IV was used as target, a low cutoff value of 0.235 optical density units (mean + 4 SD) discriminated infected and uninfected animals, with 100% sensitivity and 100% specificity. ELISA with peak IV as a target identified infected animals (even animals that had received only 10 metacercariae) within 3-5 wk of infection and subsequently throughout the rest of the 14-wk monitoring period. In Western blotting analysis, again only the antigens contained in peak IV (range 7-40 kDa, under reducing conditions) were specific for diagnosis of infected animals. These results indicate that molecular sieving of F. hepatica ESAs by this procedure is a fast, simple, reproducible way of obtaining antigens useful for serodiagnosis of sheep fascioliasis.

Animals↗
PubMed1980

Antibody responses in murine schistosomiasis and fascioliasis.

Antibody responses in mice with a primary infection of either Schistosoma mansoni or Fasciola hepatica were studied, using antigen preparations from both species. S. mansoni adult worm extracts (Ouchterlony, indrect hemagglutination) detected schistosomiasis in mice by 5-6 weeks, but did not detect fascioliasis through 12 weeks of infection. S. mansoni soluble egg antigen (Ouchterlony, enzyme-linked immunosorbent assay--ELISA) reacted with the sera of mice infected with S. mansoni by 6-7 weeks of infection; only ELISA reacted with the sera of mice infected with F. hepatica (by 3 weeks of infection). The circumoval precipitin test was specific only for mice infected with S. mansoni, being positive by the 9th week of infection. F. hepatica adult worm extract precipitated (Ouchterlony) the sera of mice with schistosomiasis (7 weeks) and fascioliasis (2 weeks).

Animals↗
PubMed1983

Use of a partially purified Fasciola gigantica worm antigen in the serological diagnosis of human fascioliasis in Egypt.

Crude extracts of Fasciola gigantica adult worms, when used as an antigen in indirect hemagglutination (IHA) and counterimmunoelectrophoresis (CIEP) tests, detected all independently diagnosed human F. gigantica and F. hepatica infections but cross-reacted with sera of patients with schistosomiasis and amebiasis. Fractionation of this crude worm extract using Sephadex G-200 chromatography demonstrated four major protein peaks. Antigen from the crest and descending portion of peak II (mol. wt. approximately 20 x 10(3)) and all of peak III (mol. wt. approximately 6 x 10(3)) were pooled and used as a source of partially purified antigen. This partially purified fraction, when used in the CIEP test, reacted with sera from patients with fascioliasis but not those from schistosomiasis or amebiasis patients, whether undiluted or concentrated fivefold, but failed to react by IHA with fascioliasis sera. It reacted with undiluted sera from all individuals passing F. gigantica eggs except one, a possibly spurious infection, and with eight of 20 sera from individuals passing F. hepatica eggs, while the remaining 12 sera became positive after fivefold concentration. It also reacted with two sera from individuals passing eggs of both Fasciola species and with five of 11 sera from individuals negative microscopically but positive serologically with the crude antigen.

Antibodies↗
PubMed1989

Detection of circulating parasite antigen in murine fascioliasis by two-site enzyme-linked immunosorbent assays.

A 2-site enzyme immunoassay was developed for the detection of Fasciola hepatica antigen in the serum of fascioliasis infected mice. The assay utilizes high titer rabbit immunoglobulins to parasite excretory/secretory antigens (FhES) as capture antibody, and also as detection antibody when linked to horseradish peroxidase (HRP) or to biotin for reaction with avidin-peroxidase. The assays were compared with a conventional (antibody detection) ELISA to determine diagnostic utility. Using mean rates of detection of fascioliasis, the HRP-based antigen capture assay diagnosed the infection at 1 week postinfection and showed that circulating antigen levels are maximal 3 weeks after infection. The earliest mean diagnosis for the antibody detection and the biotin-based antigen capture ELISAs were 2 and 3 weeks postinfection, respectively. The addition of known quantities of FhES antigens to normal mouse serum gave estimates of lower limits of detectability for the HRP- and biotin-based assays of 25 ng and 0.25 ng, respectively. Routine use of the biotin-avidin system in the antigen capture test resulted in high background activity making this method insensitive.

Animals↗
PubMed1995

Treatment of human chronic fascioliasis with triclabendazole: drug efficacy and serologic response.

Between November 1990 and March 1992, 24 asymptomatic individuals in Chile with chronic hepatic fascioliasis confirmed by the presence of Fasciola hepatica eggs in feces were treated with a single oral dose of triclabendazole (10 mg/kg of body weight) after an overnight fast. Nineteen (79.2%) of 24 patients were egg-negative two months after treatment. Three of five cases with F. hepatica in feces after the first treatment were retreated and parasitologic cure was achieved. Tolerance to the drug was excellent; none of the patients had either secondary symptoms or important alterations in levels of aspartate and amino transferases, alkaline phosphatase, or bilirubin during or after treatment. Mild eosinophilia, present in 70% of the cases, persisted at least until 60 days after treatment. An enzyme-linked immunosorbent assay (ELISA) was highly effective in diagnosis and post-treatment monitoring. Before treatment, 20 (83.3%) of 24 confirmed cases had positive test results. The test results became negative by the second month of treatment in 40% of the cured cases. This percentage increased progressively, reaching 91.3% at 12 months after therapy. In the five cases in which treatment failed, the ELISA results remained positive until the end of the follow-up period (six months). In three of these cases who accepted a second round of therapy with triclabendazole six months after the first treatment, the ELISA results became negative in all three six months after parasitologic cure and remained negative until the end of the period. Due to its efficacy, excellent tolerance, and ease of administration (a single oral dose), triclabendazole appears to be the drug of choice for chronic human fascioliasis.

Administration, Oral↗
PubMed1999

Short report: Diagnosis of human fascioliasis: detection of anti-cathepsin L antibodies in blood samples collected on filter paper.

We have developed an ELISA for the diagnosis of human fascioliasis based on the detection of IgG4 antibodies to Fasciola hepatica cathepsin LI cysteine protease. Use of this assay in the Bolivian Altiplano, a region with a high prevalence of the disease, was hampered by the reluctance of the indigenous population to provide blood. To overcome this problem, we have investigated the method of collecting small quantities of blood from the finger onto filter paper, followed by the elution of antibodies for use in the diagnostic assay. Serum samples and blood samples collected onto filter paper were obtained from 57 individuals living in the village of Cutusuma in 1987 and from 11 individuals in Chijipata in 1996. Analysis of the IgG4-ELISA results revealed that there is highly significant linear relationship (P < 0.001) between the two methods of sampling. Most importantly, a reliable diagnosis was made with the blood-filter samples from Cutusuma, which had been stored for 10 years at 40 degrees C. While some deterioration of the blood-filter samples from Cutusuma had occurred over the 10-year storage period, no deterioration occurred with the Chijipata samples, which were stored for one year. Therefore, the method of collecting blood onto filter paper should prove useful for large-scale epidemiologic studies on human fascioliasis in the Bolivian Altiplano and in other regions where this disease is prevalent.

Animals↗
PubMed1998

[The detection of antibodies, antigens and circulating immune complexes in acute and chronic fascioliasis. Preliminary results].

24 patients with fascioliasis were studied. 19 of them were in the chronic stage and 5 in acute stage. The presence of antigens and of circulating immune complexes (CIC) was detected in 100% of the patients suffering from acute fascioliasis with less than 30 days of evolution of the clinical symptoms; whereas coproantigens were present in 100% of the chronic patients. In this group it was observed a considerable number of cases with elevated levels of antibodies. 43.7% of the cases with CIC were detected by using the precipitation technique with PEG and that of Clq deviation. A highly significant correlation was found between the eggs counting and the CIC levels by both techniques. Another important correlation was established between the eggs counting and the levels of coproantigens.

Acute Disease↗
PubMed2000

Serum pretreatment with Schistosoma mansoni antigens for serological diagnosis of fascioliasis.

Cross-reactivity between Fasciola and Schistosoma often causes false positive results in serological assays for diagnosis of fascioliasis. The authors tried to reduce cross-reactivity in ELISA for diagnosis of fascioliasis by preincubation of serum samples with a mixture of Schistosoma mansoni adult worm and egg antigens. This method was evaluated using serum samples from 4 groups: 25 patients infected with Fasciola, 40 healthy controls, 113 patients infected with S. mansoni and a group of 100 patients with suspected Fasciola infection. In group with confirmed Fasciola infection, the sensitivity of ELISA was 96% without any change after serum pretreatment while, in control group, the specificity was elevated from 90% to 97.5% after serum pretreatment with S. mansoni antigens. In S. mansoni infection and suspected Fasciola infection groups, there was a highly significant reduction in number of ELISA positive cases after serum pretreatment with S. mansoni antigens (McNemar P < 0.001 for each). In logistic regression model, seroconversion showed significant dependence on presence of S. mansoni infection (P = 0.012). The probability of seroconversion was more than three times higher in S. mansoni infected individuals than in non-infected ones (Odds ratio = 3.5).

Adolescent↗
PubMed2000

Soluble adhesion molecules as a marker of disease severity in human fascioliasis.

Fasciola gigantica is one of the parasites which affect the liver by producing biliary tract lumen syndrome. Soluble adhesion molecules namely soluble intercellular adhesion molecule-1 (sICAM-1) and soluble E-selection (sELAM-1) were assayed in human fascioliasis cases with or without complications. It was found that sICAM-1 and sELAM-1 were significantly increases in complicated cases more than the control group, while only sICAM-1 was significantly increased in complicated cases more than non complicated ones. However, non of both soluble adhesion molecules were significantly elevated in non complicated cases as compared to controls suggesting that sICAM-1 is a useful marker for differentiation between complicated and non complicated cases of human fascioliasis.

Biomarkers↗
PubMed2001

Protein A immunocapture assay detecting antibodies to fluke cysteine proteinases for immunodiagnosis of human paragonimiasis and fascioliasis.

Enzyme-linked immunosorbent assays (ELISAs) which detect specific antibodies to fluke cysteine proteinases have provided good sensitivity and specificity for the immunodiagnosis of trematode diseases. To detect specific antibodies without the need for purified proteinase antigens, an immunocapture assay using Protein A was applied for the immunodiagnosis of paragonimiasis and fascioliasis. ELISA plate wells were coated with Protein A, incubated with diluted patient sera, then incubated with a preparation containing fluke cysteine proteinases, excretory-secretory (ES) products of adult Paragonimus westermani or Fasciola sp. The activity of fluke cysteine proteinases bound on the wells was measured by adding fluorogenic peptidyl substrate, Z-Phe-Arg-MCA or Boc-Val-Leu-Lys-MCA. This assay detected specific immunoglobulin G to cysteine proteinases of P. westermani and Fasciola sp. by measuring proteinase activity on the plate wells. Patient sera showed significant high values of proteinase activity when the wells were treated with the respective homologous ES products, whereas the sera had low values after treatment with the heterologous ES products. The sera of patients with other parasitoses and uninfected healthy individuals also showed low values after treatment with the above fluke ES products. Thus, Protein A immunocapture assay, which detected IgG specific for fluke cysteine proteinases, provided a high sensitivity and specificity for immunodiagnosis of paragonimiasis and fascioliasis.

Antibodies, Helminth↗
PubMed1975

Fascioliasis of livestock and snail host for Fasciola in the Altiplano Region of Bolivia.

Fascioliasis caused by Fasciola hepatica was a serious problem for sheep and alpacas in the Altiplano Region of Bolivia. In some provinces close to Lake Titicaca, the raising of sheep was forced to discontinue, because infection with the fluke made it unprofitable and almost impossible. It was proved that in the Altiplano Region, two species of freshwater snails, Lymnaea viatrix and L. cubensis var., served as intermediate hosts for F. hepatica. In some subtropical areas of Bolivia, these snails could not be found, although other Lymnaea sp. was widely distributed there. As it is possible for Lymnaea sp. to be intermediate host for the fluke, further studies are required on the identification. Acute fascioliasis of sheep occurred in the Altiplano Region principally during a period from May to July, or the dry season. In some areas, the mortality rate of infected sheep was roughly estimated as 15 to 25% annually. Contamination with Fasciola metacercariae of herbage and semi-aquatic plants grown in a swamp in one of these areas was biologically assessed, using guinea pigs. Plants of Compositae and Eleocharis sp. were contaminated most intensely and those of Senicio sp. and Vallisneria sp. carried a fairly large number of cysts, while plants of Scirpus sp. and Ranunclaceae carried only a few cysts. No signs of Fasciola infection were observed in any animal given the plants of Liliaceae.

Animals↗
PubMed2001

Fascioliasis in Vietnam.

A confirmed diagnosis of human fascioliasis was rare in Vietnam until 1978 when two cases were reported in humans. Since 1997, we have confirmed 500 cases of human fascioliasis. The majority of cases come from the central provinces of Vietnam: Da Nang, Quang Ngai, Binh Dinh, Phu Yen and Khanh Hoa. Patients were treated in hospitals in Ho Chi Minh City. All had high peripheral blood eosinophilic counts (16-70%) and positive serology with Fasciola gigantica antigen with positive titers of 1/1,600 to 1/12,800. We are unsure whether this represents an endemic pattern of disease or whether improved specific laboratory tools now facilitate better diagnosis. It is also possible that with changes in environmental factors and in the number and breeds of herbivorous domestic animals, Fasciola is increasing in frequency and easily contaminates the food.

Adult↗
PubMedDate not supplied

Specific IgG avidity in acute and chronic human fascioliasis.

The detection of IgG avidity in sera is potentially useful in the diagnosis of acute and chronic infection. We studied IgG avidity in 31 patients with fascioliasis, with the aim of evaluating the clinical application of this test to confirm the diagnosis of incubating cases and to distinguish between acute and chronic cases. Of the 31 cases, 13 were incubating and had a mean avidity index of 57.28 +/- 5.79%. The 18 chronic cases had an avidity index of 68.80 +/- 8.92%. The difference was highly significant. We conclude that IgG avidity is a reliable means of identifying the stage of fascioliasis and suggest a cut-off point of 59.90% to distinguish between acute and chronic infection.

Adolescent↗
PubMed2002

Serum IgM. Does it relate to the level of chronicity in fascioliasis?

Serum IgM and IgG specific to human fascioliasis were reported to monitor prepatent and patent human infection respectively. Recently an assay measuring the antigen binding avidity of IgG antibodies was proved to be of value in identification of the stage of fascioliasis. The present study investigates the level of serum IgM in relation to avidity IgG, among acute and chronic Fasciola patients (using ELISA). Specific IgM antibody was detected in all acute and in 88% of chronic cases. IgM serum level was significantly higher in acute cases. IgG antibody was found in both acute and chronic cases but with no significant difference between the two groups. Regarding IgG avidity, it was found that acute cases had low IgG avidity index whereas chronic cases had significantly higher avidity. Four chronic patients had high IgM level and high IgG avidity. The egg counts as well were relatively high. The possibility of reinfection was considered in those cases.

Acute Disease↗