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PubMed1994

Complement depletion in rats infected with Fasciola hepatica: in vivo and in vitro studies.

The action of Fasciola hepatica on rat complement was evaluated by measuring the haemolytic activity. Extracts from metacercariae and eggs and the excretory and secretory products of adult flukes reduced the classical pathway of complement-mediated lysis by normal rat sera in a protein dose-dependent manner. Whole metacercariae produced the same effect. In vitro incubation of newly excysted juveniles and intact eggs with normal rat serum had no detectable effect on serum complement activity by either the alternative or classical pathway. During the course of rat fasciolosis a depletion in the classical pathway of complement-mediated lysis was demonstrated particularly 6-9 weeks post-infection; the alternative pathway of complement-mediated lysis was not affected.

Animals↗
PubMed1998

IgG isotype specific immune response in rats infected with Fasciola hepatica.

Antibody responses (IgG1, IgG2a, IgG2b, and IgG2c subclasses) against Fasciola hepatica L. in rats infected with metacercariae were analysed by ELISA. Animals of group 1 (R-1) remained infected throughout 21 weeks, and rats of group 2 (R-2) received a single oral dose of triclabendazole (Fasinex 10%, Ciba-Geigy) at 4 weeks after infection. Group C (R-C) consisted of rats left intact which served as uninfected controls. All IgG subclasses increased during the first weeks after infection, but when triclabendazole was administered IgG1 and IgG2b diminished markedly. IgG2c showed a different profile to the other antibodies evaluated, increasing earliest and decreasing profoundly from the 9-11th week after infection (w.a.i). The infected-untreated rats produced higher titres of antibodies than the rats of R-2, and these differences were statistically significant (p < 0.05) in all subclasses evaluated except in IgG2a titres.

Animals↗
PubMed1997

[The influence of treatment on the course of experimental Fasciola hepatica infection and weight gains in sheep].

The course of Fasciola hepatica infection was studied by the coproscopical methods in two farms of Southern Poland. The extensity of infection reached between 84 to 94 per cent. The experimental studies has been done on 54 lambs at the average weight of 21.50 kg. The lambs were divided in 3 groups with 18 lambs in each. In the first group were non-infected animals; second group was infected with 200 metacercaria and treated with Fasinex (10 mg/#kg body weight) 6 weeks after infection. The third group was infected with 200 metacercaria, but has not been treated with Fasinex. The best results were obtained in the first group. In this group the weight gain was about 4 kg (14.7 per cent) higher comparing with the group third (infected, non-treated) group. The weight gain of group two (infected and treated) was 2.42 kg (11.0 per cent) higher as in group third group.

Animals↗
PubMed1976

The stimulation of daughter redia production during the larval development of Fasciola hepatica.

In snails maintained at 20 degrees C rediae of Fasciola hepatica merge from sporocysts from 11 days after infection onwards. The number of mother rediae rises steadily thereafter until at least 40 days after infection. Daughter rediae are seldom observed in mother rediae dissected from snails maintained at 20 degrees C. Their production can, however, be stimulated by subjecting the snail host to starvation, to low, and to high temperature shocks. The parasite is susceptible to stress from immediately after infection for about 16 days, when maintained at 20 degrees C. In general, the more extreme the shock, the greater is daughter redial production. Increasing the length of the period of stress from 12 h up to 9 days does not increase the production of daughter rediae, nor does repeated on/off cold shocks or continuous maintenance at 10 degrees C. Daughter rediae develop more rapidly than cercariae and leave the mother rediae several days earlier. There is no evidence that presence of daughter rediae coincides with the suppression of cercarial production. The findings are discussed with reference to possible mechanisms by which parasite development might be controlled.

Animals↗
PubMed1999

Oviposition of Lymnaea truncatula infected by Fasciola hepatica under experimental conditions.

Experimental infections of Lymnaea truncatula by Fasciola hepatica (one, two, or three miracidia per snail) were carried out under laboratory conditions to analyze the oviposition of infected snails and determine the characteristics of their egg masses. In the infected snails from the three groups, egg-laying steadily decreased until week 4 postexposure and stopped afterward until the end of the experiment, except for the cercaria-shedding snails from the one-miracidium group, for which low numbers of egg masses were noted between weeks 9 and 12. In uninfected snails the number of egg masses decreased until week 4 and remained low during subsequent weeks. At weeks 11 and 12 postexposure the natality rate was 98.9% in controls, 56% in the cercaria-shedding snails from the one-miracidium group, and 59.2-68.5% in uninfected snails. In the cercaria-shedding snails from the one-miracidium group the restoration of reproduction activity after week 8 may be explained by a lower parasite burden in these snails than in those from the two- and three-miracidium groups.

Animals↗
PubMed1975

Acquired immunity to Fasciola hepatica in splenectomised rats.

Significant resistance to a second infection with Fasciola hepatica was obtained in splenectomised and sham operated rats (P less than 0.001, 78 and 76 per cent respectively) when compared with singly infected splenectomised or sham operated control groups. For both the stimulating and challenge infections, there were no significant differences in the number of flukes recovered from rats that had been splenectomised as compared to those receiving the sham operation. Thus, splenectomy did not significantly affect the ability of rats to develop an acquired immunity to F hepatica, nor were fluke recoveries from a particular schedule of infection significantly affected by the presence of absence of a spleen. It is concluded that the presence of a spleen is not necessary for the development of acquired immunity to F hepatica in the rat.

Animals↗
PubMed1978

Experimental cross infections of Fasciola hepatica in lambs and calves.

The effects of experimental infections with Fasciola hepatica of ovine and bovine origin in homologous and heterologous hosts and in uninfected controls were compared; groups comprised 5 animals each. The effects of the infections were monitored by biweekly determinations of packed cell volumes (PCV), serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), gamma glutamyl transpeptidase (GGT), serum iron, bilirubin levels, alkaline phosphatase (AlP) and total serum protein levels. Infected animals showed changes in SGOT, SGPT and GGT activity levels, and GGT activity levels, and infected lambs showed changes in PCV and AlP. However, no no significant differences in these serum levels between infected host groups were attributable to fluke strain. At necropsy, calves infected with ovine and bovine strains on an average had about the same number of flukes, but lambs infected with a high dose of the bovine strain on the average had nearly twice the number of flukes as those infected with ovine strain. Weight gains did not differ within host groups; liver damage was extensive in all infected animals. On the basis of these experiments, the pathogenicity of the ovine and the bovine strains of F. hepatica appears to be the same.

Alanine Transaminase↗
PubMed1978

The metabolic profile of adult Fasciola hepatica obtained from rafoxanide-treated sheep.

Sheep infected with adult Fasciola hepatica were drenched with rafoxanide. At 4, 8, 16 and 24 h after drenching the sheep were killed and the flukes removed, washed and rapidly frozen in liquid nitrogen. The content of key metabolites in the fermentation pathway were determined and compared with those in control F. hepatica, whose hosts were not treated with rafoxanide. Rafoxanide decreased glycogen, malate, NADH and ATP levels. The level of other metabolites in the pathway increased for the first 8-16 h after rafoxanide treatment. The marked decrease in ATP and glycogen, and the increase in total [NAD+]/[NADH] and [oxaloacetate]/[malate], together with the changed content of other metabolites, led to the conclusion that the mode of action of rafoxanide against F. hepatica in vivo is by uncoupling oxidative phosphorylation.

Adenosine Diphosphate↗
PubMed2001

Metacercarial aggregation in Digenea (Fasciola hepatica and Paramphistomum daubneyi): environmental or species determinism?

Metacercarial aggregation of Fasciola hepatica and Paramphistomum daubneyi was studied under experimental conditions to determine if the formation of these aggregates was influenced by environmental factors, or it was a characteristic of trematode species. This process was studied using the confinement of infected snails on the bottom of Petri dishes (diameter, 14 cm) for 3 days. The formation of metacercarial aggregates of F. hepatica was not significantly modified by environmental factors such as intensity and duration of lighting, quality and volume of water. Metacercariae of F. hepatica were more numerous on the Petri dish walls and 63.9% of them constituted aggregates. In contrast, most metacercariae of P. daubneyi were found on the Petri dish bottoms and 78.3% of them were isolated or in groups of two metacercariae each. The mean number of metacercariae per aggregate ranged from 6.7 to 12.2 in the case of F. hepatica, and from 2.7 to 4.5 in the case of P. daubneyi. However, these mean numbers were independent of the site of cercarial attachment. The tendency of cercariae to form metacercarial aggregations was a characteristic of F. hepatica and was species determined.

Animals↗
PubMed1979

[Fasciola hepatica in Denmark. A survey on 15 years diagnostic examination on bovine faeces samples (author's transl)].

The results of routine diagnostic examinations of 10,540 bovine faeces samples for eggs of Fasciola hepatica during the period 1963-1977 are evaluated. 0n an average, 10.4% of the samples were positive. The frequency of positive samples was highest among samples from cows (14.4%) lowest among samples from calves (1.5%). Of samples from heifers 10.1% were found positive. Calculated on a monthly basis, the percentage of positive samples showed a clear seasonal variation, reaching a peak in the period December-March. During the period 1973-1977, the annual percentage of positive samples showed a rather pronounced declining tendency, indicating a decrease in the occurrence of bovine Fasciola hepatica infection in Denmark.

Age Factors↗
PubMed1980

Fasciola hepatica: influence of thymus function on the liver pathology in mice.

Liver pathology during experimental Fasciola hepatica infection was compared in thymus-deficient (nu/nu) mice and in thymus-bearing (+/+) mice, (back ground strain to nu/nu mice) and thymus-reconstituted nu/nu mice. Thymus deficiency resulted in unrestricted liver damage during the migration of the parasite in the liver. In nu/nu mice the migratory tracks were infiltrated by very few eosinophils and mononuclear cells, haemorrhages and liver cell necroses were extensive. Portal area reaction and bile duct hyperplasia were depressed. Regeneration of liver tissue, parenchymal and periportal fibrosis and plasma cell proliferation were reduced. The possible causal mechanisms involved in fluke induced liver pathology are discussed. It is suggested that a thymus-dependent cellular reaction modulates the tissue damage produced by F. hepatica.

Animals↗
PubMed1982

Antigen sharing between Fasciola hepatica and human erythrocytes.

Several lyophilized bovine and ovine liver flukes (Fasciola hepatica) extracts were examined for the presence of blood group like antigens using an automated haemagglutination inhibition procedure. This sensitive method allowed the detection of A, B, H. Lea, Leb, P1, and Pk group specific substances. Their immunogenicity was confirmed in anti-Fasciola hyperimmunized rabbits. Immunofluorescent probes showed that these substances are located on cell membranes of the tegumental syncytium and caeca epithelium cells of the fluke. These data were discussed along with current concepts concerning, the origin, the consequences ant the biological significance of the occurrence of blood group like substances in parasites.

ABO Blood-Group System↗
PubMed1989

Fasciola hepatica:Sp2/0 (helminth:myeloma) hybridoma expressing parasite antigen.

Cells from adult Fasciola hepatica were fused with cells from a murine BALB/c myeloma Sp2 line. The hybrid cells were grown in HAT (hypoxanthine, aminopterin, and thymidine) medium, cloned and subcloned, and shown to express parasite antigen for 1 year after fusion. Expression of parasite antigen was demonstrated by the following: 2 histogram flow cytometric analyses, in which a population of hybrid cells in the population of 7 month cultured hybrid cells showed 57% more fluorescence when treated with an anti-F. hepatica serum followed by anti-rabbit immunoglobulin G coupled to fluorescein isothiocyanate as compared with the same hybrid cells washed and treated with normal rabbit serum; Sp2 myeloma cells treated with an anti-F. hepatica serum or normal rabbit serum followed by fluorescein-labeled anti-rabbit IgG had the same negative fluorescence; BALB/c mice immunized with PBS-washed cells from a subclone of these hybridomas developed anti-F. hepatica antibodies (shown by the Falcon assay screening test enzyme-linked immunosorbent assay); and antibodies recognized an F. hepatica antigenic polypeptide of 57,000 Mr in a Western immunoblot. These helminth:myeloma hybrids expressed murine host markers, further confirming the hybrid nature of this cell line. F. hepatica cells alone, like their Sp2 fusion partners, die in HAT supplemented medium by 9 days of culture. F. hepatica:Sp2 hybridomas have been grown continuously in HAT medium for greater than 1 year.

Animals↗
PubMed1997

Presence of very high prevalence and intensity of infection with Fasciola hepatica among Aymara children from the Northern Bolivian Altiplano.

Coprological studies of school children from four communities in the Northern Bolivian Altiplano were carried out in order to estimate the prevalences and intensities of Fasciola hepatica infection. Single stool specimens were collected at random from 558 school children (308 boys and 250 girls) aged 5-19 years old. Nineteen different parasite species (13 protozoan and six helminths) were detected. Of the children examined, 98.7% (96.5-100%) presented infection with at least one parasite species. The mean prevalence of 27.6% by Fasciola hepatica (range, 5.9-38.2%) was the highest not only with respect to the helminth species found in the Northern Bolivian Altiplano but also among the fasciolosis prevalences reported in children in other parts of the world to date. Prevalences were significantly different among the communities surveyed and was significantly higher in the 9-12 years age group. There were, however, no significant differences between sexes. Among the 154 children presenting F. hepatica eggs in stools, intensities ranged from 24-5064 eggs per gram of faeces (epg), with arithmetic and geometric means of 474 and 201 epg, respectively. Significant differences in mean egg output were detected between communities, sexes and age groups. Individual fasciolosis infections coexisting with other pathogenic parasite species (Entamoeba histolytica and/or E. dispar, Giardia intestinalis, Balantidium coli, Dientamoeba fragilis, Cryptosporidium sp., Hymenolepis nana, Taenia spp., Trichuris trichiura, Ascaris lumbricoides and Enterobius vermicularis) were detected. A significant positive association with F. hepatica was only found in the case of G. intestinalis. This coprological study not only verifies the existence of high prevalences of F. hepatica among humans in the Northern Bolivian Altiplano, but also demonstrates the need to expand the Southern boundaries of this high endemic zone to include the Southeastern region of Lake Titicaca.

Adolescent↗
PubMed2004

Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR.

From complete mitochondrial DNA sequence of Fasciola hepatica available in Genbank, specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used for diagnosis of infected Lymnaea columella by F. hepatica during the pre-patent period simultaneously with another pair of primers which amplified the internal transcribed spacer (ITS) region of rDNA from L. columella in a single Multiplex-PCR. The amplification generated a ladder band profile specific for F. hepatica. This profile was observed in positive molluscs at different times of infection, including adult worms from the trematode. The Multiplex-PCR technique showed to be a fast and safe tool for fascioliasis diagnosis, enabling the detection of F. hepatica miracidia in L. columella during the pre-patent period and identification of transmission areas.

Animals↗
PubMed2004

A novel Fasciola hepatica saposinlike recombinant protein with immunoprophylactic potential.

A member of the Fasciola hepatica saposinlike/NK-lysin protein family with lytic activity on human peripheral blood mononuclear cells and erythrocytes was recently described. The current study was designed to test the immunoprophylactic potential of this protein termed FhSAP-2 against infection with F. hepatica in rabbits. Two doses of 50 microg of recombinant FhSAP-2 (rFhSAP-2) emulsified in TiterMax were injected subcutaneously on the dorsal surface of 4 rabbits at 2-wk intervals. Four weeks after the second immunization, the rabbits were infected orally with 25 F. hepatica metacercariae. Four non-immunized-infected rabbits were used as controls. An enzyme-linked immunosorbent assay revealed high levels of antibodies to both rFhSAP-2 and F. hepatica excretory-secretory antigens by 2 wk after the first immunization, which were always significantly higher in immunized-infected rabbits than in control-infected rabbits. On the completion of the trial, vaccinated rabbits had 81.2% less flukes than controls. Moreover, F. hepatica egg counts in feces, as well as in bile collected from the gall bladders from vaccinated animals, were lower, 83.8 and 73%, respectively, compared with controls. The vaccinated rabbits also had significantly lower amounts of parasite antigen in stool and bile samples than controls. Last, evaluation of macroscopic liver lesions revealed that the rabbits vaccinated with rFhSAP-2 had milder lesions than the infected-control rabbits. These findings support the hypothesis that this novel rFhSAP-2 protein has immunoprophylactic potential against fascioliasis in rabbits including antifecundity and antipathology effects. This is the first report on experimental vaccination of rabbits against F. hepatica with a purified, defined, recombinant protein related to a member of the saposinlike protein family.

Analysis of Variance↗
PubMed1995

Biochemical analysis of recombinant glutathione S-transferase of Fasciola hepatica.

Four cDNAs encoding GST (rGST1, rGST7, rGST47 and rGST51) of Fasciola hepatica were expressed in Escherichia coli and the rGST proteins purified for biochemical analyses. The rGST proteins are 95% pure as indicated by Coomassie staining of proteins separated by SDS-PAGE. Molecular sieving by HPLC infers that, like the native protein, the rGST proteins form homodimers under non-denaturing conditions. The rGST proteins are recognised by antisera raised to the native GST of F. hepatica. All four rGST proteins from F. hepatica actively conjugate glutathione to the universal substrate, 1-chloro-2,4-dinitrobenzene. The activity of the rGSTs was also measured for substrates which have been shown to have partial specificity for the Alpha, Mu or Pi classes of mammalian GSTs (trans-4-phenyl-3-buten-2-one, ethacrynic acid), for substrates known to be products of lipid peroxidation (trans-2-nonenal, trans,trans-2,4-decadienal) and for epoxy-3-(p-nitrophenoxy)-propane (EPNP), a known substrate for the theta class of GST. No rGST were active with EPNP. rGST47 and 51 showed activity with the other four substrates. rGST7 was active with three substrates whereas rGST1 showed relatively low activity with all substrates except trans,trans-2,4-decadienal. The sensitivity of the rGST activity to inhibition by the GST inhibitors triphenyltin chloride and bromosulphophthalein also varied among the rGSTs with rGST1 showing a 800-fold difference in sensitivity between the inhibitors. These results show that F. hepatica expresses a family of GST isoenzymes which exhibit unique substrate and inhibitor profiles.

Amino Acid Sequence↗