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PubMed2005

The contamination of wild watercress with Fasciola hepatica in central France depends on the ability of several lymnaeid snails to migrate upstream towards the beds.

As most natural watercress beds in central France are located upstream of the permanent habitats of two lymnaeid species, Galba truncatula and Omphiscola glabra, field investigations were made from 1999 to 2004 on 67 beds to determine why the contamination of watercress with Fasciola hepatica is irregular over time in these sites, while definitive hosts, especially lagomorphs, are regularly found infected around them. Snails are able to migrate upstream in winter and spring towards the beds, and a 4-year survey demonstrated the existence of annual variation in the colonization of these sites by snails. In the 45 beds irregularly contaminated with F. hepatica over time, 37.7-62.2%, according to the year, were not populated by lymnaeids, in spite of the presence of snail populations downstream. G. truncatula was found alone in 8.8-13.3% of sites and was the first colonizing snail in 24.3-33.3% when the two lymnaeid species successively settled in these waterholes. The colonizing ability of O. glabra was more limited, as it was observed alone in 2.2% of beds and was the first colonizing snail in only 2.2-20% of them. The distances covered by these snails significantly increased with increasing migration time. After migration, a few overwintering snails (3.8% of G. truncatula and 6.8% of O. glabra) are able to colonize the beds, but their numbers decreased when the distance of migration was greater. O. glabra migrated more quickly and reached more watercress beds than G. truncatula. However, this did not influence the natural infections of snails, which were more frequent in the F1 of G. truncatula. The variability observed in the colonization of beds by snails might explain the regular or irregular contamination of wild watercress by metacercariae of F. hepatica. One of the factors which may explain this variability is the occurrence of showers in spring, so that a few snails are able to reach watercress beds after their upstream migration.

Animals↗
PubMed2004

Expression of interleukin 4, interleukin 4 splice variants and interferon gamma mRNA in calves experimentally infected with Fasciola hepatica.

Interleukin 4 (IL-4) is expected to play a dominant role in the development of T helper (Th) 2 cells. Th2 immune responses with expression of relatively large amounts of interleukin 4 (IL-4) but little interferon gamma (IFN-gamma) are characteristic for chronic helminth infections. But no information is available about IL4 expression during early Fasciola hepatica (F. hepatica) infections in cattle. Therefore, we investigated F. hepatica specific IL-4 and IFN-gamma mRNA expression in peripheral blood mononuclear cells (PBMCs) from calves experimentally infected with F. hepatica. Cells were collected prior to infection and on post-inoculation days (PIDs) 10, 28 and 70. Interestingly, PBMCs responded to stimulation with F. hepatica secretory-excretory products (FhSEP) already on PID 10 and expressed high amounts of IL-4 but not of IFN-gamma mRNA suggesting that F. hepatica induced a Th2 biased early immune response which was not restricted to the site of infection. Later in infection IL-4 mRNA expression decreased whereas IFN-gamma mRNA expression increased slightly. Isolated lymph node cells (LNCs) stimulated with FhSEP and, even more importantly, non-stimulated LN tissue samples indicated highly polarized Th2 type immune responses in the draining (hepatic) lymph node, but not in the retropharyngeal lymph node. During preliminary experiments, two splice variants of bovine IL-4 mRNA, boIL-4delta2 and boIL-4delta3, were detected. Since a human IL-4delta2 was assumed to act as competitive inhibitor of IL-4, it was important to know whether expression of these splice variants of bovine IL-4 have a regulatory function during an immune response to infection with F. hepatica. Indeed, IL-4 splice variants could be detected in a number of samples, but quantitative analysis did not yield any clue to their function. Therefore, the significance of bovine IL-4 splice variants remains to be determined.

Alternative Splicing↗
PubMed2001

Immunoprophylaxis against Fasciola hepatica in rabbits using a recombinant Fh15 fatty acid-binding protein.

Previous studies of ours have demonstrated that a recombinant protein (Fh15) related to fatty acid-binding proteins did not induce significant protection in rabbits challenged 2 or 4 wk postimmunization over nonimmunized controls. In the current study, rabbits were immunized with Fh15 and challenged with Fasciola hepatica metacercariae 12 and 20 wk later. In the current study in which longer lag periods for challenge infection after the second immunization were used, worm burden reductions compared to adjuvant controls were a significant 43% and 76%, respectively. Importantly, rabbits immunized with Fh15 had significant numbers of immature flukes, 66% in the 12-wk period and 84% in the 20-wk lag period as compared to controls. In addition, liver lesions were clearly diminished in the vaccinated rabbits. Enzyme-linked immunosorbent assay absorbance values showed that immunized rabbits developed high antibody levels to Fh15 from 8 wk after the first immunization and did not increase after challenge. These results suggest that a recombinant F. hepatica molecule related to fatty acid-binding proteins induces protective (worm burden reductions), anti-fecundity (immature flukes), and anti-pathology (less liver lesions) effects in rabbits and may serve as a model for the immunoprophylaxis of fascioliasis.

Animals↗
PubMedDate not supplied

Role of the prosegment of Fasciola hepatica cathepsin L1 in folding of the catalytic domain.

The N-terminal propeptides of cysteine proteinases play regulatory roles in the folding and stability of their catalytic domains, as well as being potent and highly specific inhibitors of their parental mature enzymes. Cysteine proteinases play a major role in the biology of the parasitic trematode Fasciola hepatica; in particular, this organism secretes significant amounts of cathepsin L enzymes. The isolated propeptide of F. hepatica cathepsin L1 functioned as a chaperone for the mature enzyme in renaturation experiments. A double point mutation (N701/F721) within the GxNxFxD motif of the propeptide affected its conformation and markedly decreased its affinity for the mature enzyme. When this mutation was introduced into preprocathepsin L1 expressed in yeast, the secretion of active enzyme dropped dramatically. However, significant enzyme activity was recovered from the culture supernatants after denaturation and renaturation in the presence of native propeptide. Thus, the variant prosegment gave rise to an enzyme with altered conformation, which could be refolded to the active form with the assistance of the native propeptide.

Amino Acid Sequence↗
PubMed2005

Risk factors for Fasciola hepatica infection in children: a case-control study.

We conducted a case-control study to clarify the risk factors for human fascioliasis in the Northern Peruvian Altiplano. Cases (n=61) were individuals who were diagnosed as having chronic fascioliasis by faecal and/or serologic (Fas2-ELISA) studies. Controls (n=61) had negative serologic and coprological results for Fasciola hepatica. We identified an association between fascioliasis and four variables (40 analysed): the habit of drinking alfalfa juice (OR=4.5; 95% CI 1.8-11.1; P<0.001); familiarity with aquatic plants (OR=4.3; 95% CI 1.8-10.6; P<0.001); dog ownership (OR=5; 95% CI 1.7-15.1; P=0.002); and raising more than five sheep (OR=0.3; 95% CI 0.1-0.8; P=0.01). According to clinical presentation and laboratory studies, dizzy spells (P=0.01), history of jaundice (P=0.01), peripheral eosinophilia (P=0.005) and Ascaris lumbricoides in stools (P=0.001) were associated with fascioliasis. The principal exposure factor for F. hepatica infection was drinking alfalfa juice. In conclusion, we suggest that human fascioliasis in Peru should be suspected in patients from livestock-rearing areas, who present with recurrent episodes of jaundice and who have a history of consumption of alfalfa juice or aquatic plants, or who have eosinophilia.

Adolescent↗
PubMed1998

Immunohistochemical study of the local immune response to Fasciola hepatica in primarily and secondarily infected goats.

The distribution of CD2, CD4, CD8, gamma/delta T-lymphocytes, B-cells and IgG lambda-light chain (lambda-IgG) containing cells were analysed in the inflammatory infiltrate associated to hepatic lesions and gallbladder (HL), and in hepatic lymph nodes (HLN) of goats primarily and secondarily infected with Fasciola hepatica. In the HL, CD2 and CD8 T-cells were more numerous (p=0.01) in secondarily rather than in primarily infected goats, whereas CD4 T-lymphocytes were less numerous than CD8 and showed no significant change in both groups. The ratio CD4/CD8 was 0.66 and 0.39 for primarily and secondarily infected goats, respectively. In contrast, in the HLN, CD4 were more numerous than CD8 T-cells, the ratio CD4/CD8 was 2.0 in control, 1.5 and 1.3 in primary and secondary infections, respectively. Gamma/delta T-lymphocytes were scarce in the HL and moderate in the HLN of both primarily and secondarily infected animals. B-cells (IgM+, lambda-IgG+ or CD79+) varied from scarce or moderate in the HL to abundant in the HLN, where CD79+-cells were mainly located in lymphoid follicles and IgM and IgG in plasma-cells of the medullary cords, suggesting an intense local humoral immune response. However, this response did not prevent the hepatic damage in secondarily infected goats, in which hepatic lesions were more severe than in primarily infected ones.

Animals↗
PubMed2003

Cloning and expression of the major secreted cathepsin B-like protein from juvenile Fasciola hepatica and analysis of immunogenicity following liver fluke infection.

The functions of the cathepsin B-like proteases in liver flukes are unknown and analysis has been hindered by a lack of protein for study, since the protein is produced in small amounts by juvenile flukes. To circumvent this, we isolated and characterized a cDNA encoding the major secreted cathepsin B from Fasciola hepatica. The predicted preproprotein is 339 amino acids in length, with the mature protease predicted to be 254 amino acids long, and shows significant similarity to parasite and mammalian cathepsin B. Only one of the two conserved histidine residues required for cathepsin B exopeptidase activity is predicted to be present. Recombinant preproprotein was produced in yeast, and it was shown that the recombinant proprotein can undergo a degree of self-processing in vitro to the mature form, which is active against gelatin and synthetic peptide substrates. The recombinant protein is antigenic in vaccinated rats, and antibodies to the protein are detected early after infection of rats and sheep with F. hepatica. The kinetics of the response to cathepsin B and cathepsin L after infection of sheep and rats confirm the temporal expression of these proteins during the life cycle of the parasite.

Amino Acid Sequence↗
PubMed1998

Observations on the mechanism of eggshell formation in the liver fluke, Fasciola hepatica.

A mechanism for eggshell production in Schistosoma mansoni has been proposed (Wells & Cordingley, 1991), and suggests that the release of eggshell protein globules from the vitelline cells occurs under alkaline conditions within the ootype followed by their subsequent fusion to form the eggshell. Fusion and tanning of these components produces eggshell which autofluoresces. The present study was carried out to determine whether a similar process operates in Fasciola hepatica. A number of drug treatments were used to disrupt key steps in the maturation of vitelline cells. Treatment with the calcium ionophore lasalocid (1 x 10(-5) M) led to the premature release of eggshell globules from the vitelline cells but not their fusion. Incubation in monensin (1 x 10(-6)M), a sodium ionophore and ammonium chloride (NH4Cl) (5 x 10(-2) M), a weak base, resulted in the premature fusion of eggshell protein globules within the vitelline cells and premature tanning of the eggshell protein material. The copper-containing enzyme, phenol oxidase, is thought to be involved in the tanning process during the production of eggs. Diethyldithiocarbamate, (DDC, 1 x 10(-3) M) is a phenol oxidase inhibitor and treatment with this compound, in combination treatments with monensin and NH4Cl, prevented fusion of the vitelline cell globules and tanning of the shell protein material. The results of the study suggest that the mechanism for eggshell formation in F. hepatica is similar to that proposed for S. mansoni and may be common to other trematodes as well.

Ammonium Chloride↗
PubMed1982

Loss of substrate binding capacity of the hepatic microsomal cytochrome P-450 in Fasciola hepatica infected rats: toxicological implications.

Experimental fascioliasis in the rat is responsible for a dramatic decrease in the drug-metabolizing ability of the hepatic monooxygenase system. The present investigation, through a spectroscopic study of hexobarbital interaction with microsomal cytochrome P-450 and in vitro and in vivo studies of hexobarbital metabolism in the rat, demonstrates that this decrease is due to an alteration in the structure of the hemoprotein (loss of substrate binding capacity of cytochrome P-450 followed by denaturation). These results might be responsible for a decreased safety margins for those flukicidal agents that are detoxified by the monooxygenase pathway, and might explain the accumulation problems frequently associated with chemotherapy of Fasciola hepatica.

Alcohol Oxidoreductases↗
PubMedDate not supplied

[Acute pancreatitis complicating biliary distomatosis caused by Fasciola hepatica in a patient with a choledochal diverticulum].

A 43 years old man presented with acute pancreatitis following 13 years of mild grade hepatic colitis and angiocholitis. The absence of visible gall stone evocated the possibility of millithiasis. The operation discovered a big and hard head of the pancreas and an hypertrophied choledocal mucous membrane and the microscopic analysis of bladder's bile, ova of Fasciola hepatica. A control opacification on the 7th day revealed an adult fluke and a small diverticulum which were not visible on the roentgenogram at the time of the operation. Seven infusions of 60 mg de dehydroemetin permitted to eliminate the fluke with only a peak of fever. The diagnostic, pathophysiologic and therapeutic implications are discussed.

Acute Disease↗
PubMedDate not supplied

Specific tyrosine phosphorylation in response to bile in Fasciola hepatica and Echinostoma friedi.

Protein tyrosine phosphorylation (PY) is a well-known signalling mechanism which is also involved in host-parasite interactions. Despite its transcendence, PY has been poorly studied in parasitic helminths. The aim of this study is to examine the effect of bile salts on the PY pattern in parasitic trematodes. Two distinct adult models were analysed: Echinostoma friedi, of intestinal habitat, and Fasciola hepatica, naturally inhabitant of host biliary channels. Our results show that bile salts induce specific and distinct protein PY in both trematode species, indicating that this signalling process seems to be also involved in host-trematode relationships.

Animals↗
PubMed1980

Enzyme assay for detecting resistance to reinfection with the liver fluke Fasciola hepatica in rats.

The kinetics of the release of a liver enzyme, glutamate dehydrogenase, into the serum of Fasciola hepatica infected rats was studied. The results support previous work that has shown that barriers to reinfection with this parasite are met before any pronounced liver damage takes place. The presence of this enzyme in serum can be used as an indicator of resistance to reinfection.

Animals↗
PubMed1998

Relative concentrations of heavy metals in the parasites Ascaris suum (Nematoda) and Fasciola hepatica (Digenea) and their respective porcine and bovine definitive hosts.

The concentrations of lead and cadmium determined by electrothermal atomic absorption spectrometry were significantly higher in the liver and kidney, respectively, of pigs than in their intestinal nematode parasites Ascaris suum. There was no clear pattern in the distribution of lead within the ascarids, but cadmium concentrations were highest in the intestine. A parallel investigation of cattle naturally infected with the liver fluke, Fasciola hepatica, revealed interesting differences. Although the cadmium content of F. hepatica was considerably lower than that in the tissues of cattle, the concentration of lead in the digenean was on average 172, 53 and 115 times higher than in the muscle, kidney and liver of the host. Furthermore, there was a significant positive correlation between the weight of individual F. hepatica and their lead burden. The lack of appreciable heavy-metal accumulation in A. suum is consistent with results for the nematode Anguillicola crassus in fish. However, although lead concentrations in the liver fluke F. hepatica were considerably elevated above host tissue levels, the degree of heavy-metal accumulation was relatively low when compared to that of acanthocephalans and cestodes of fish.

Animals↗
PubMed1994

Comparison of the migrations of Fasciola hepatica sporocysts in Lymnaea truncatula and other related snail families.

The migration and definitive localization of sporocysts in the snail body were studied in four species (Aplexa hypnorum, Lymnaea glabra, L. truncatula, Physa acuta), each of which was exposed to a single miracidium and killed during a period ranging from the 1st to the 5th day postexposure. When penetration occurred through the mantle, tunnels were found mainly in the mantle or kidney in Lymnaea adults and also extended to the foot in many physidid adults and in newborns of the four species. The sporocysts preferentially localized in the zone surrounding the kidney and heart beginning on the 4th day in Lymnaea spp. and disappeared in Aplexa and Physa snails on the 3rd day. When penetration occurred through the foot and extended inconstantly to the mantle according to the snail species involved. Sporocysts were encountered in the foot as well as the mantle or the zone surrounding the kidney and heart. The significance of these results is discussed in relation to the snail host and the entry site used by the Fasciola hepatica miracidium.

Animals↗
PubMedDate not supplied

[Extrahepatic obstructive jaundice caused by Fasciola hepatica].

A 58 year old female patient with diabetes manifested 30 years ago is reported. She had digestive disorders since 20 years ago, and a diagnosis of cholecystopathy had been established. Furtherly, she had jaundice, acholia and pruritus; this picture remitted spontaneously. During the next year she had again jaundice, epigastric pain and vomitings; this picture also remitted with home rest. Fifteen days prior to her admittance she had again jaundice, epigastralgia extending to the back, vomitings and pruritus, as well as a diabetic uncontrol. She was admitted though bile ducts were not roentgenographically explored as a result of iodine allergy. A Fasciola hepatica was intraoperatively found.

Cholestasis, Extrahepatic↗