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PubMed2000

Molecular cloning and expression of Cu/Zn-containing superoxide dismutase from Fasciola hepatica.

The cytosolic superoxide dismutase (SOD) of Fasciola hepatica, a causative agent of fascioliasis, was purified and characterized. The enzyme consists of two identical subunits, each with an apparent molecular mass of 17.5 kDa. An analysis of the enzyme's primary structure and inhibition studies revealed that the enzyme is a copper/zinc-containing SOD (Cu/Zn-SOD). The enzyme activity was relatively stable in a broad pH range, from pH 7.0 to 10.0, and the enzyme showed maximum activity at pH 7.5. This enzyme also displayed strong antigenicity against sera of bovine and human subjects with fascioliasis. The SOD gene fragment was amplified by PCR with degenerate oligonucleotide primers derived from amino acid sequences conserved in the Cu/Zn-SODs of other organisms. An F. hepatica cDNA library was screened with the SOD gene fragment as a probe. As a result, a complete gene encoding the Cu/Zn-SOD was identified, and its nucleotide sequence was determined. The gene had an open reading frame of 438 bp and 146 deduced amino acids. Comparison of the deduced amino acid sequence of the enzyme with previously reported Cu/Zn-SOD amino acid sequences revealed considerably high homologies. The coding region of the F. hepatica Cu/Zn-SOD was cloned and expressed in Escherichia coli. Staining of native polyacrylamide gel for SOD activity of the expressed protein revealed SOD activity that was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide. This means that the presence of the recombinant fusion protein is indicative of Cu/Zn-SOD. The expressed protein also reacted with sera of bovine and human subjects with fascioliasis, but it did not react with sera of uninfected bovine and human subjects.

Amino Acid Sequence↗
PubMed1998

Excretory-secretory antigenic components of adult Fasciola gigantica recognized by infected human sera.

The immunogenic components of Fasciola gigantica excretory-secretory (ES) products were revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technic using sera from patients with F. gigantica infection, from patients with clinical suspected fascioliasis, from patients with other illness and from healthy adults. By SDS-PAGE, it was found that the ES products comprised more than 6 polypeptides. Immunoblotting analysis revealed 12 components which were strongly recognized by fascioliasis antisera. These antigenic components had a molecular mass ranging from less than 14.4 to 38 kDa. One antigenic band of 27 kDa was found to give a consistent reaction with fascioliasis antisera (100% sensitivity and 98% specificity). The present findings suggest that the 27 kDa components are sensitive and specific for the diagnosis of human F. gigantica infection.

Adult↗
PubMedDate not supplied

Intra-specimen and day-to-day variations of Fasciola egg counts in human stools.

Intra-specimen and day-to-day variations of Fasciola egg counts in stools were investigated for 16 cases of established fascioliasis. For each case six Kato slides from a single stool sample were examined daily for 5 consecutive days. The results indicated the presence of significant intra-specimen variations in more than one-third of the examined series, while the inter-specimen variation was almost negligible. The sensitivity of the Kato-Katz test for diagnosing Fasciola infection with three Kato slides from the same specimen or on different days ranged from 96.0%-99.1%. The examination of three Kato smears from a single stool specimen, which is more feasible in field studies, would give an accurate diagnosis of fascioliasis. Used as such, the Kato-Katz technique is highly sensitive in the diagnosis of fascioliasis.

Adult↗
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↗
PubMed1990

A study of a restricted programme of strategic dosing against Fasciola hepatica with triclabendazole.

A restricted programme of strategic dosing with triclabendazole was used to treat a sheep flock with severe clinical fascioliasis. After five years the percentage of ewes passing fluke eggs was reduced from 49 per cent to less than 1 per cent, without the reappearance of clinical fascioliasis. It is suggested that this programme offers an efficient and practical means of controlling fascioliasis.

Animals↗
PubMed1979

Use of immunologic techniques to detect chemotherapeutic success in infections with Fasciola hepatica. II. The enzyme linked immunosorbent assay in infected rats and rabbits.

An enzyme-linked immunosorbent assay (ELISA) was developed using microtiter plates for the immunodiagnosis of fascioliasis in rats and rabbits using extracts of adult worms partially purified by gel filtration chromatography using Sephacryl S-200. Partial purification was necessary to eliminate cross-reactivity with antisera having antibodies to schistosomes. Soft polyvinyl plates clearly gave superior results over hard polystyrine plates. Titers rose by 4 weeks of infection in rats with fascioliasis, by 6 weeks in the case of rabbits, and remained high through at least 12 and 28 weeks, respectively. Titers drop rapidly when animals are successfully treated with a fasciolicidal drug at 4--6 weeks of infection. The results show that the ELISA can be employed for the serodiagnosis of fascioliasis in rats and rabbits and is useful for the prediction of chemotherapeutic success.

Animals↗
PubMed1998

[Detection and quantification of Fasciola hepatica antigens in sheep].

The diagnosis of ovine fascioliasis hepatica is presented by a sandwich ELISA with the use of monoclonal antibody ES78 previously reported for the diagnosis of bovine fascioliasis. The assay was applied to faeces of 40 sheep infected with Fasciola hepatica, 88 sheep with other parasitary infections tested by coproparasitology and 100 caprologically negative animals. Antigens of excretion-secretion of Fasciola hepatica were detected in 38 of the 40 sheep with fascioliasis and in none of the negative animals or with other parasitosis. The results show that ES78 sandwich ELISA, as in bovines, is fast, simple and useful for the diagnosis of the active infection caused by Fasciola hepatica among bovines.

Animals↗
PubMed1994

[Status of parasitic infection diagnosed by surgical biopsy in Kwangju and Chollanam-do].

In order to know the species and frequency of human parasitic infection diagnosed by biopsy, 149 cases (0.18%) of parasitic infection were reviewed, which were selected from 80,947 biopsied materials submitted for routine histopathological examination during a period of 10 years from 1980 to 1989 at Department of Pathology, Chonnam National University Hospital. They consisted of 112 cases of cysticercosis, 17 paragonimiasis, 7 clonorchiasis, 4 amebiasis, 1 sparganosis, 1 enterobiasis, 1 anisakiasis, and 1 fascioliasis respectively. Based on morphological preservation of cysticercus, they could be divided into mild (20.2%), moderate (40.4%), and severe (39.4%) degeneration. Except 2 cases biopsied at the lungs, 15 cases of ectopic paragonimiasis were located at abdominal cavity (8 cases) and central nervous system (7 cases). One case of intrahepatic fascioliasis was observed. This is the 13th human fascioliasis reported in Korea. From the above results, the frequency of parasitic infections found in biopsied specimens was on the decrease as the year passed by, but biopsy is very useful diagnostic method on tissue parasites such as cysticercosis and ectopic paragonimiasis.

Adolescent↗
PubMed1995

Antifasciolic efficacy of indigenous plant drugs: kalonji, shahterah and karanjwa in buffaloes.

A study was carried out to evaluate antifasciolic effect of powdered plant drugs including Kalonji (Nigella sativa seeds), Shahterah (Fumaria parviflora aerial parts) and Karanjwa (Caesulpinia crista seeds) in buffaloes. During the search for the infected animals, prevalence of fascioliasis in the random samples was 33.1 +/- 0.7% but in symptomatically suspected cases it came to be 64 +/- 3.2%. The drug trial results showed that all the three drugs possess significant efficacies against fascioliasis. Their highest doses tested produced highly significant (P<0.001) decrease in EPG counts on 15th days. Maximum antifasciolic efficacy, judged on the basis of % EPG count reduction was shown by Shahterah (93.2 +/- 0.5%) which was followed by Karanjwa (89.7 +/- 1.7%) and Kalonji (88.2 +/- 0.4%). No visible side effects were produced by any of these plant drugs. Single oral treatment with 25 mg/kg of Kalonji or 60 mg/kg of Shahterah and 40 mg/kg of Karanjwa exerts highly significant antifasciolic efficacies on the day 15 after treatment. From these data, it is conceivable that at these dosages the drugs are sufficiently potent and safe to treat fasciola infection in buffaloes. However, further chemical and pharmacological studies would decide the exact mechanism(s) of action, the active principles contained and the real worth of these indigenous drugs for the treatment of fascioliasis in the ruminants.

Journal Article↗
PubMed1996

Parasitic causes of hepatomegaly in children.

Three hundred children with hepatomegaly were selected. They were subjected to full clinical and laboratory examinations. Also serum samples were examined to detect IgG using ELISA against SEA, chromatography purified hydatid cyst antigen, commercially available Toxoplasma antigen, partially purified adult Fasciola antigen and second-stage larvae Toxocara canis antigen. IFAT was used to detect IgG against Toxoplasma and T. canis. A commercially available IHAT kit for leishmaniasis was used. Based on immunological assays, 125 cases were suffering from various parasitic infections. Thirty cases with schistosomiasis (10%), 26 cases fascioliasis (8.7%), 18 toxocariasis (6%), 35 toxoplasmosis (11.7%), 3 cases hydatidosis (1%) and 13 cases mixed parasitic infections. No parasitic causes could be found in 175 cases (58.3%). Moderate or marked hepatomegaly favours the presence of schistosomiasis. Whereas, most cases with other parasites and those with non-parasitic infections fall in the category of mild hepatic enlargement. There was no associated splenomegaly in cases with Fasciola, Toxocara, hydatid disease and/or the non-parasitic group. Most of hepatomegalic cases with non-parasitic causes were found to be associated with fever (88.5%). Fever was found in nearly 50% of cases with either Toxoplasma or Toxocara infections. Mild eosinophilia was found in all cases with parasitic causes. Only 24 cases of non-parasitic group (13.7%) had easinophilia. Moderate and high eosinophilia were found in cases with fascioliasis and toxocariasis. Cases with fascioliasis had a statistically significant increase in enzymes activities specially alkaline phosphatase. It was concluded that parasitic infections should be considered as an important cause of liver enlargement in children. Serological methods using purified antigenic fractions are an important tool for diagnosis.

Child↗
PubMed1996

Parasitic infections presenting as prolonged fevers.

Over two successive years, out of 187 cases of fevers of undetermined origin (FUO) admitted to Abbassia and Embaba Fever Hospitals, 30 (16%) cases proved to be of parasitic origin. Ten within normal subjects were taken as controls. Complete blood picture, repeated stool examination, rectal snip by transparency technique, ELISA for specific IgM antibodies for S. mansoni, indirect haemagglutination test for S. mansoni, Fasciola, hydatid, amoebic liver abscess and toxoplasmosis, indirect fluorescent antibody test for toxoplasmosis and abdominal ultrasonography were performed whenever indicated. Cases comprised 8 (26%) acute S. mansoni, 7 (24%) acute fascioliasis, 3 (10%) hydatid cyst, 8 (26%) amoebic liver abscess, 2 (7%) toxoplasmoisis and 2 (7%) malaria cases. The clinical picture of acute S. mansoni and acute fascioliasis were similar in the form of prolonged fever, diarrhea, hepatomegaly and leucocytosis with high eosinophilia. Serology (ELISA and IHAT) was essential in differentiating them. Abdominal ultrasonography is an easy, sensitive, cheap, non-invasive technique aiding in the diagnosis of amoebic liver abscess, liver hydatid cysts and fascioliasis but again serology was essential in differenting them. Toxoplasmic lymphadenitis mimic the clinical picture of infectious mononucleosis. Serology (monospot test, IHAT, IFAT) clinched the diagnosis. Malaria cases presented atypically by gastrointestinal manifestations and hepatic affection. Diagnosis was by positive blood smears.

Diagnosis, Differential↗
PubMed1994

Fasciola hepatica: mebendazole and thiabendazole pharmacokinetics in sheep.

Pharmacokinetics of the two anthelmintic drugs mebendazole and thiabendazole were determined in sheep before and 4, 8, 13, 19, and 25 weeks after an infestation of animals by an oral administration of 150 metacercariae of Fasciola hepatica. The parasitic pathology was ascertained by the increase in plasma enzyme activities of glutamate dehydrogenase and gamma-glutamyltransferase. After oral administration of mebendazole (25 mg.kg-1), the parent drug and especially its reduced metabolite were present in plasma of animals. A significant 1.5- to 2.7-fold increase in the mean residence time occurred by Weeks 13 to 25 postinfection. This change was related to decreases in both the elimination from the pharmacokinetic compartment representing the reduced metabolite and the area under the curve of plasma metabolite concentration versus time. A 59% decrease in MBZ reduction was demonstrated in liver microsomes prepared from 12-week-infected sheep. This reductase activity was characterized by NADPH dependency and a pH peak activity of 6.0 and was competitively inhibited by daunomycin. In sheep receiving a 50 mg.kg-1 oral dose of thiabendazole, fascioliasis provoked only decreased plasma concentrations of the metabolite 5-hydroxythiabendazole by Weeks 4 to 25 postinfection. This change parallels an increase in urinary excretion of free metabolite but this is of minor significance in the general fate of the drug because of the prevalence of excretion as conjugates. In summary, fascioliasis appears to have more of an effect on the pharmacokinetics of mebendazole, a drug intensively metabolized by the liver into a metabolite present at high concentrations in the plasma of animals and humans.

Administration, Oral↗
PubMed1979

Ovine facioliasis (Fasciola gigantica) on the Ahmadu Bello University farm.

Outbreaks of fascioliasis (Fasciola gigantica) in sheep were observed on the Ahmadu Bello University farm during the end of the dry season in 1972-74 and in 1978. During 1972 especially the pregnant ewes were affected. Pre- and post-mortem exmaination of the affected sheep showed typical signs of acute fascioliasis; the number of flukes in the liver ranged between 77 and 239. The infections were acquired when the animals grazed near the man-made lake on the farm. A limited snail survey revealed the presence of Lymnaea natalensis with the highest incidence of snails as well as of larval Fasciola infections in the snails during the beginning of the dry season. There was an indication that the control measures taken after 1974, i.e. not allowing the animals to graze around the lake especially during the wet season, prevented the occurrence of outbreaks in 1975-77.

Animals↗
PubMed1984

Ovine internal parasitism in the Yemen Arab Republic.

Internal parasitism was studied in five flocks in different regions of the Yemen Arab Republic. Ewes and lambs were faeces sampled from the spring of 1977 to the spring of 1978 and helminth-free tracer lambs were subsequently run with three of the flocks from October 1978 to November 1979. The findings indicated that ovine helminthiasis and fascioliasis may be of importance with the following regional variations: Haemonchosis all the year round in the Tihammah coastal plain. Haemonchosis and trichostrongylosis in the periparturient period in the Southern Uplands and Western Foothills. Fascioliasis and nematodiriasis in the Inter-Montane Plains of the Highlands. Possible control regimes are suggested for further investigation.

Animals↗
PubMed1987

Fasciola hepatica: liver enzymes in rats and interaction with chemical inducers.

Adult male rats were sorted into control and infected groups, the latter receiving an oral dose of 20 metacercariae of Fasciola hepatica. In Weeks 3 and 6 after infection, some rats received phenobarbital or 3-methylcholanthrene which induced drug metabolizing enzymes. The parasitic pathology was ascertained by clinical observation of the rats and at autopsy. Hepatic microsomal cytochrome P-450 content was significantly decreased in infected rats compared to untreated phenobarbital treated groups. In all infected rats, the simultaneous increase in cytosolic calcium and decrease in cytosolic glutathione corresponded to oxidative cell injury occurring in the course of fascioliasis. Both arylamine acetyltransferase (EC 2.3.1.5.) and glutathione transferase (EC 2.5.1.18) activities were decreased in all newly infected and 6 week infected groups. Fascioliasis did not alter the substrate related uridine diphosphate glucuronosyltransferase activities (EC 2.4.1.17) of any rat group.

Animals↗
PubMed1991

Hepatic disposition of organic anions in rats infested with Fasciola hepatica.

This study was undertaken to investigate the effects of subclinical fascioliasis at various stages of its development (3, 6, and 11 weeks after infestation with an oral dose of 20 metacercariae of Fasciola hepatica) on the hepatobiliary transport of organic anions in rats. Basal bile acid secretion was not significantly modified in infested rats as compared to the controls but there was a significantly increased bile flow in the 11-week group. In all infested rats, maximal biliary excretion and cumulative excretion of bromosulphthalein was significantly reduced, although there was a tendence to recovery in the 11-week infested group. Decreased biliary secretion was accompanied by increased plasma and liver concentrations of bromosulphthalein. Glutathione S-transferase activity was significantly reduced by Weeks 3 and 6 postinfestation. Liver glutathione concentration was significantly reduced in all infested rats. Maximal excretion of dibromosulphthalein, a compound not biotransformed in the liver, fell by Week 6 postinfestation and recovered a normal value thereafter. These results indicate that effects of experimentally induced fascioliasis on hepatic disposition of organic anions depend upon the stage of its development and that are a consequence of both metabolism and transport related factors.

Animals↗
PubMed1983

Levels of type I and type III collagen in the bile duct of rats infected with Fasciola hepatica.

Collagen types and their levels were compared between bile ducts from Fasciola infected rats and bile ducts from uninfected animals. Both collagen types I and III were shown to be increased in infected animals but, levels of type I increased less than type III. These results indicate that fascioliasis produces changes in the collagen composition of the bile duct that are similar to those produced in cirrhosis of the liver and other pathologic conditions including wound healing. Such observations suggest that a study of the chronology of collagen deposition in fascioliasis might provide information on the sequence of molecular events which result in bile duct hyperplasia.

Animals↗
PubMed1984

The effect of sex on seasonal variation in single and double infection of cattle in Sierra Leone by Dicrocoelium hospes and Fasciola gigantica.

The incidence of dicrocoeliasis (61.8%) is higher than that of fascioliasis (25.5%) and their peaks do not occur at the same time of the year. The peak for the former occurs in the dry season while that of the latter occurs in the rainy season. The incidence of dicrocoeliasis and fascioliasis in female cattle was found to be higher than that of male cattle. Double infection was higher in female (14.7%) than in male hosts (9.6%).

Animals↗