Metacercariae Of Fasciola Hepatica Research Articles

The Effects of Experimental Fascioliasis on Bilirubin Metabolism in the Rat

The effects of subclinical fasciolosis at various stages of its development on bilirubin metabolism were investigated in rats. Studies were carried out at 3 (initial lesions), 6 (parenchymal degeneration), and 11 weeks (biliary localization of the parasite) after infection with an oral dose of 20 metacercariae of Fasciola hepatica. Both bile flow and bile acid secretion were significantly reduced by Weeks 3 and 6 postinfection, while a significantly increased bile flow was found in the Ii-week group. Serum concentration of unconjugated bilirubin was elevated at 3-6 weeks postinfection and that of conjugated bilirubin and the esterified/total bilirubin ratio increased in all infected rats. Biliary excretion of bilirubin mono- and diconjugates was maximally increased by 3-6 weeks, remaining elevated at 11 weeks. No significant change in the biliary diconjugate/monoconjugate ratio was found in any of the groups. UDP-glucuronosyltransferase activity was reduced by Weeks 3 and 6. This was accompanied by a significantly lowered liver cytochrome P450 concentration and an elevation in hepatic heme oxygenase activity. Blood hemoglobin concentration was significantly reduced and reticulocyte count increased by 6 and 11 weeks. Our results indicate that enhanced bilirubin production, cholestasis, and a reduced conjugating activity could be responsible for the changes in plasma and bile pigment composition during the acute stage of fasciolosis. Hemolysis and a high bilirubin excretion are maintained when the parasite migrates into the biliary ducts.

Fasciola-Hepatica: Liver Microsomal Membrane Functions in Host Rat

Changes in different microsomal membrane functions were measured in the liver of rats 3, 6, or 9 weeks following an oral infection with 20 metacercariae of Fasciola hepatica. The parasitic pathology noted at autopsy was accompanied by increased levels in both plasma aspartate aminotransferase (EC 2.6.1.1) and microsomal γ-glutamyltransferase (EC 2.3.2.2). Heme oxygenase activity of microsomes was significantly decreased by Weeks 3 and 6 postinfection and this decrease correlates with those of total microsomal cytochrome P450 and certain P450-dependent monooxygenase activities, namely, benzphetamine demethylation, ethoxycoumarin deethylation, and benzopyrene hydroxylation. Microsomal epoxide hydrolase (EC 3.3.2.3) was only altered 6 weeks after the infection. During the early stages of the parasitism, there were decreases in both microsomal calcium uptake and calcium ATPphosphohydrolase activity (EC 2.6.1.1), whereas membrane fluidity, estimated by the order parameter S, was lower in the infected rats than that in the controls. These alterations could be related to the already described increase in liver cytosolic calcium or lipid peroxidation which occurs in experimental fascioliasis.

Parasitic gastritis in a llama ( Lama glama) associated with inhibited larval Teladorsagia spp. (Nematoda: Trichostrongyloidea)

A 7-year-old female llama was experimentally infected with 500 Fasciola hepatica metacercariae then turned out onto pasture also grazed by domestic sheep. The llama was necropsied approximately 22 weeks post infection. Routine examination of the gastrointestinal tract revealed diffusely coalescing, umbilicated nodules covering the caudal one-fifth of the third stomach compartment. Microscopically, the mucusa was irregularly thickened. Numerous nematode larvae were present in glandular lumens, often extending to the base of the glands. Where nematodes were most numerous, decreased numbers of parietal cells, attenuation of glandular epithelium and increased collagen within the lamina propria were evident. Of a total of 6510 adult nematodes in the third stomach compartment, 51% were Teladorsagia spp., 47% were Trichostrongylus axei, and 2% were Camelostrongylus mentulatus. Except for C. mentulatus, three sheep that had grazed with the llama and were necropsied at approximately the same time, harbored the same nematode species. The total number of larvae in the llama was 56 710, with more than 97% being ostertagiine early fourth-stage larvae. Ostertagiine larvae were also present in the sheep. Based on the composition of the adult populations in the llama and sheep, the larvae were considered to be species of Teladorsagia. Additionally, the larger numbers of early fourth-stage larvae indicated that the development of Teladorsagia spp. in the llama is inhibited.

Antipyrine, erythromycin and oxytetracycline disposition in experimental fasciolosis

The effects of fasciolosis on drug disposition were studied by administration of antipyrine, erythromycin and oxytetracycline to sheep and cattle. Fasciolosis was produced by administration of 200 or 400 metacercariae (MC) of Fasciola hepatica to sheep and 500 MC to cattle. The disease was subsequently confirmed by determination of plasma glutamate dehydrogenase and γ-glutamyl transferase and identification and quantitation of mature flukes in the liver at necropsy. Acute or subacute fasciolosis in sheep was accompanied by a significant decrease in the elimination rate constant (β) and increase in the elimination half-time (t 1/2) for antipyrine and erythromycin when compared with controls or infected sheep which had been treated with the anthelmintic luxabendazole. An increase in apparent volume of distribution (Vd) was seen only for erythromycin in sheep given 400 mc. There were no changes in the disposition of oxytetracycline in sheep with either acute or subacute infection and no effects on disposition of the three test drugs in chronically infected sheep. With early chronic disease in calves, only the disposition of oxytetracycline was affected; not that of antipyrine or erythromycin.

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.

Differential inhibition of rat hepatic glutathione S-transferase isoenzymes in the course of fascioliasis

The effects of a subclinical fascioliasis at various stages of its development (at week 3, 6 and 9 after infection by oral administration of 20 metacercariae of Fasciola hepatica) in rats were determined on the activity of enzymes involved in liver metabolism of glutathione and on the subunit pattern of cytosolic glutathione S-transferase. 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 by week 3 and 6 post-infection. Not correlatively, the catalytic activities of glutathione S-transferase towards 1-chloro-2,4-dinitrobenzene and 1,2-dichloro-4-nitrobenzene were significantly lowered in last stages of the experimental fascioliasis (by week 6 and 9 post-infection). These decreases were correlated to that of subunit 1 as determined by means of high-performance liquid chromatography of cytosolic proteins whereas subunit 6 could also be decreased. Fasciolasis did not alter cytosolic glutathione, glutathione reductase and glutathione peroxidase activities or plasma glutathione S-transferase activity accepting 1-chloro-2,4-dinitrobenzene as the substrate.

Cellular reactions in the small intestine of rats after infection withFasciola hepatica

Groups of five rats each were infected with metacercariae of Fasciola hepatica according to two experimental procedures. In the first experiment, they received 20 metacercariae 0, 1, 2 or 3 times at intervals of 4 weeks. In the second, both the inoculation dose and the interval between doses were varied. After the animals had been killed, the small intestine was prepared for histology and tissue sections were stained for the detection and quantification of mucosal mast cells (MMCs), eosinophils and goblet cells. The density of eosinophils and MMCs increased significantly with each reinfection in the first experiment; in the second, the MMC reaction reached a peak at 4 weeks post-infection (p.i.), whereas the eosinophils peaked at 4 and 17 weeks p.i. In addition, the reaction of these cells depended on the infectious dose. In both experiments, goblet cells reacted only weakly. These findings thus clearly reflect a considerable intestinal response against migrating juvenile F. hepatica. The pattern of these defence reactions in the small intestine suggests some similarities with the mechanisms of antibody-dependent cell-mediated cytotoxicity that have been shown to take place during Schistosoma mansoni reinfection.

Decrease in albendazole sulphonation during experimental fascioliasis in sheep

1. The in vivo S-oxidation of albendazole was measured from the pharmacokinetic profile of albendazole sulphoxide and sulphone determined in young male sheep receiving oral albendazole (1.9 mg/kg). Studies were carried out before, and each month after, oral infestation by 150 metacercariae of Fasciola hepatica. 2. Parasitic pathology was ascertained by clinical observation of animals, and the increase in plasma antibodies directed against liver flukes. 3. Rate of conversion of sulphoxide to sulphone and rate of sulphone elimination, were respectively decreased by 47% and 87% at week 8 post-infection, whereas significant increases in the area under plasma sulphone concentrations versus time curve and mean residence time, occurred 4-12 weeks following the infestation. 4. A 58% decrease in albendazole sulphonation was demonstrated in liver microsomal preparations obtained from 8-week-infected sheep, while there was no change in the FAD-directed sulphoxidation of albendazole. 5. The transient impairment of albendazole sulphonation could be related to the decrease in liver microsomal cytochrome P450-dependent monooxygenases observed in sheep with a similar parasitic pathology.

Pharmacokinetics of ampicillin and pentobarbital in the course of subclinical fascioliasis in sheep

Pharmacokinetics of two common veterinary drugs, ampicillin and pentobarbital, were determined in sheep before and four, eight, 12,17 and 21 weeks after infestation of animals by an oral administration of 150 metacercariae of Fasciola hepatica. The parasite infestation was ascertained by clinical observation of the animals. The pharmacokinetics of ampicillin were not significantly affected by the liver parasitism but the disposition of pentobarbital changed. A significant increase in elimination half-life (around 180 per cent), volume of distribution (130 per cent) and mean residence time (154 to 170 per cent) was observed in sheep infected by the parasite for four to 12 weeks. In these animals, duration of narcosis caused by pento-barbital was prolonged 1·8-fold. The results suggested that both reduced elimination of pentobarbital and impaired distribution of the drug would be responsible for the prolonged duration of narcosis in infected animals.

The efficacy of triclabendazole (fasinex ®) against immature and adult fasciola hepatica in experimentally infected cattle

Eighteen Chinese cattle were experimentally infected with metacercariae of Fasciola hepatica and randomly assigned to 6 groups. Five groups of cattle were treated with a single oral dose of triclabendazole at a dose rate of 12 mg kg −1. At necropsy, the reduction in fluke burden compared with the untreated group was 85, 99.6, 99.8, 100 and 100% for cattle treated 2, 6, 8, 12 and 16 weeks after infection, respectively. Data are also presented on body weight changes during the experimental period and on serum γ-GT activity in cattle from selected groups. Triclabendazole is considered to be safer and more efficacious than currently available fasciolicides in China.

The epidemiology of fasciolosis in Papua New Guinea.

Fasciola hepatica metacercariae were present on pasture throughout the year at the trial site. The highest infection rate in trial sheep occurred in the first year after the beginning of the wet season, in December/January, when contamination of the pasture was maintained by other sheep. Variation in rates of infection in consecutive years was linked to differences in grazing pressure and to a change from sheep to cattle as accompanying grazing stock. A minimum precipitation of 125 mm over 4 consecutive weeks appears to be necessary for infected snails to move from their normal habitats and to contaminate wet pasture with cercariae. Infection of sheep can occur during a dry season if pasture has permanent seepage sites or swampy areas where infected snails can remain active and continue to liberate cercariae.

Comparative incidence of experimental fascioliasis on corticosteroid pharmacokinetics in sheep

1. The kinetics of intravenously administered prednisone (0.5 mg/kg) and methylprednisolone hemisuccinate (4 mg/kg) were compared in two groups of four young male sheep before and regularly after experimental infestation with 150 metacercariae of Fasciola hepatica and 5 or 7 weeks following a flukicidal treatment administered post-infection at week 25. 2. Parasitic pathology was ascertained by clinical observation and by the increase in plasma antibodies directed against liver flukes. 3. The disposition of prednisone was altered with a significant decrease of plasma clearance occurring from eleven weeks after infection onwards. This change was parallel to an increase in the mean residence time of prednisone and of its major metabolite (prednisolone). 4. The same parasitic burden provoked only a slight decrease in the elimination half-life of methylprednisolone alcohol when its hemisuccinate ester was administered.

Incidence of a subclinical fascioliasis on antipyrine clearance and metabolite excretion in sheep.

1. The pharmacokinetics of i.v. antipyrine (25 mg/kg) used as a model compound, were determined in young male sheep, before and each month after an oral infestation by 150 metacercariae of Fasciola hepatica, and 8 weeks following a flukicidal treatment. 2. The parasitic pathology was ascertained by the clinical observation of animals and the increase in plasma antibodies directed against liver flukes. 3. A significant decrease in the total plasma clearance of the drug occurred by week 4 to 16, and a 1.7 fold increase in mean residence time occurred by week 12 post-infection. 4. Urinary excretion of antipyrine metabolites was determined before and 8 weeks following the infestation. 4-Hydroxyantipyrine was the major urinary metabolite and its excretion was decreased by 30% in infected sheep, whereas there was no change in the excretion of norantipyrine, 3-hydroxymethylantipyrine or unmetabolized drug. 5. It is concluded that the impairment of antipyrine clearance in the course of fascioliasis could be related to the decrease in liver microsomal cytochrome P-450-dependent mono-oxygenases observed in sheep with a similar parasitic burden.

Pharmacokinetics of sulphobromophthalein, lidocaine and indocyanine green in the course of subclinical fascioliasis in sheep

The pharmacokinetics of three drugs proposed for the assessment of liver function: sulphobromophthalein (BSP), lidocaine and indocyanine green (ICG) were determined in sheep four, eight, 12, 16 and 24 weeks after their infestation by an oral administration of 150 metacercariae of Fasciola hepatica. The disposition of BSP was altered by a significant decrease in its total plasma clearance from eight weeks after infection onwards. This change could be related to the low values of the elimination rate constant consistent with a reduced liver cytosolic conjugation of the dye to glutathione. Lidocaine pharmacokinetics were unaffected by parasitism and the only effect on ICG was an increased volume of distribution consistent with the liver hypertrophy caused by the subclinical fascioliasis.

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.

Heterologous resistance to Fasciola hepatica conferred upon rats by passive transfer of serum from different breeds of sheep

Two experiments compared the protection against oral challenge with 20 metacercariae of Fasciola hepatica conferred on rats by intraperitoneal injection of serum from three breeds of sheep infected with F. hepatica (Barbados Blackbelly, St. Croix, Florida Native). Experiment 1 used serum from sheep 5–6 months of age following two infections of 250 metacercariae each, while Experiment 2 utilized serum collected from the same sheep at 10–11 months of age following either a primary (first exposure) or challenge (after two previous exposures of 250 metacercariae each) infection with 500 metacercariae. Similar numbers of flukes were recovered from rats given either immune or nonimmune (control) serum from each breed of sheep in Experiment 1. In Experiment 2, rats given serum from infected St. Croix sheep had significantly fewer flukes than rats given either control or immune serum from Barbados Blackbelly or Florida Native sheep. There was no significant correlation of fluke counts between individual serum donors (sheep) and serum recipients (rats).

Early migration of radiolabeled larvae of Fasciola hepatica in the mouse

Metacercariae of Fasciola hepatica were labeled with 75Se by exposure of infected Lymnaea columella to 75SeM, 20 μCi/snail, in 5 ml of water. Seventy-two mice were inoculated with 50 75Se-labeled metacercariae. At intervals after infection, the distribution of metacercariae in gut, body cavities and liver was studied by compressed organ autoradiography, and a variant of this technique adapted to gut content and body cavity washes. Ag° focus counts from autoradiograms expressed as percentage of actual exposure level were subjected to probit analysis. Estimates of the average departure time of radiolabeled maritas from gastrointestinal tracts and average arrival time into the abdominal cavities were 12.2 and 12.5 h, respectively. The peak accumulation of maritas in abdominal cavities occurred at 24 h post-inoculation. A 7.7-h discrepancy between estimated average departure time from abdominal cavities (92 h) and average arrival time at livers (99.7 h) was attributed to experimental error.

The uptake and distribution of radioselenium in the larvae of Fasciola hepatica and its snail host Lymnaea columella

Radiolabeled metacercariae of Fasciola hepatica were obtained in vivo by incubating infected Lymnaea columella snails with 20 μCi radioselenomethione ( 75SeM) per snail in 5 ml of water for 5 h, or in vitro by incubating a batch of unlabeled F. hepatica metacercariae with 75SeM for 24 h. Radioassay showed that only 5% of the 75SeM was incorporated into maritas (juvenile flukes) from the in vivo labeled metacercariae. The inner cyst wall of in vivo labeled metacercariae contained 46% of the total activity, of which 21% was dissolved in the excysting medium. The outer, tan-colored cyst wall contained 49% of the radioactivity. Through diffusion/attachment, maritas from in vitro labeled metacercariae could occasionally be labeled with 0.4% of the total radioactivity. However, the activity was lost after inoculation into the body of mice. The outer and inner cyst walls of in vitro labeled metacercariae contained 92% and 7.6%, respectively, of the total activity. Microautoradiography demonstrated that 75SeM was evenly distributed in the body of marita and the cyst wall of inner and outer layers from an in vivo labeled metacercaria. A 9 × 4 μm rectangularly-shaped aggregate of Ag° grains was present on the outer periphery of the inner cyst wall. Microautoradiography of in vitro labeled metacercariae demonstrated a significant concentration of Ag° grains on the cyst walls. The ventral plug contained fewer Ag° grains per unit area compared to the other portion of the inner cyst wall. Uptake and distribution of 75SeM in the snail host were also studied. It appeared that rediae and cercariae tended to concentrate the label in the foot, the mantle and the digestive gland. Little or no radioactivity was present in the areas where F. hepatica larvae were not found.

The susceptibility of the goat to Fasciola hepatica infections

Mixed breed goats were infected with metacercariae of Fasciola hepatica and the resulting worm burdens were quantitated after primary and secondary exposure of the goats to the parasite. Mean length and width of the parasite recovered after all primary exposures were 1.91 ± 0.2 cm and 0.91 ± 0.2 cm, respectively. A mean of 71.8 ± 5.9% of the flukes were recovered from all of the primary infections. In the secondary infections, the mean length and width of the flukes from the physically smaller population was 0.88 ± 0.27 cm and 0.53 ± 0.19 cm, respectively. A mean of 67 ± 6.7% of the flukes were recovered from this secondary infection. It appears that the goat is susceptible to challenge infections with F. hepatica and that its response to this infection is much like that of sheep.

Induction of drug metabolizing enzymes in the liver of rats infested with Fasciola hepatica.

Adult, male rats were infected with 20 metacercariae of Fasciola hepatica given orally, other rats were left untreated. Five weeks after infestation, some animals received phenobarbitone, 3-methylcholanthrene, beta-naphthoflavone or Arochlor 1254, to induce liver drug metabolizing enzymes. Fascioliasis provoked decreases in aminopyrine N-demethylase, aniline hydroxylase, the mutagenic activity of cyclophosphamide and cytochrome P-450 concentration in untreated or phenobarbitone or Arochlor pretreated rats. In contrast, cytochrome b5, NADPH cytochrome c reductase, ethyoxycoumarin O-deethylase and the enzymatic activation of ethidium bromide were not affected by fascioliasis whatever pretreatment was given. Fascioliasis decreased liver drug metabolizing enzymes which were specifically induced by both phenobarbitone and Arochlor, this could be due to either the specific action of toxic excretions of flukes or to the particular localization of tissue damage within the liver lobule.