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PubMed1996

Fasciola hepatica infection in sheep: changes in liver metabolism.

Several aspects of liver function during infection with Fasciola hepatica were examined in sheep four weeks after infection and compared with the changes observed in infected rats. Previously reported respiratory abnormalities in mitochondria isolated from the left lobe of the liver of infected sheep were characterised further. Evidence is presented that the respiratory lesion is located in the mitochondrial electron transport chain and that the aberrant respiratory behaviour is not associated with an increase in nonesterified fatty acids and the depletion of mitochondrial phospholipids, as is the case in the rat. Microsomal membranes, which have also been shown to be depleted of phospholipids in the fluke-infected rat liver, showed no such changes in the sheep. However, in common with the rat, a substantial loss of cytochrome P450 was recorded in microsomes prepared from the left lobe, and the glycogen content of the left lobe was found to be less than 50 per cent of control values. No change was observed in glucose 6-phosphatase activity. All these changes were localised effects, confined to areas of fluke infiltration.

Animals↗
PubMedDate not supplied

Lymphocyte and cytokine responses of young cattle during primary infection with Fasciola hepatica.

An experiment was designed to characterise the early immune responses of cattle given a trickle infection of the trematode Fasciola hepatica. Lymphocyte responses and IFNgamma production following stimulation with Fasciola antigens were measured. Animals were sequentially slaughtered at weeks 5, 7, 9 and 11 post-infection. Results demonstrated responses to fluke antigens and cytokine production (IFNgamma) from weeks 2-5 post-infection. These findings may suggest an early stimulation of IFNgamma production by peripheral blood lymphocytes following infection with F hepatica in cattle.

Animals↗
PubMed2000

Identification and localization of a schistosome-associated fucosyllactose determinant expressed by Fasciola hepatica.

A Biomphalaria alexandrina-derived lectin (BaSII), of proven specificity to a Schistosoma mansoni-associated fucosyllactose [(Fuc alpha1-2) Gal beta1-4 Glc] determinant, was employed to investigate the putative antigenic cross-reactivity between Schistosoma mansoni and Fasciola hepatica in terms of this structurally-defined oligosaccharide sequence. BaSII affinity column chromatography of extracts of adult worms metabolically radiolabelled with 35S-methionine and analysis by two-dimensional gels established the expression of the fucosyllactose determinant in multiple copies among heterogeneous, acidic glycoproteins synthesized by adult Fasciola hepatica. Direct fluorescence microscopy revealed that determinant-bearing glycoproteins were localized to the external glycocalyx and perikarya of the tegument as well as the epithelial lining of the intestinal caeca and vitelline ducts and glands. Determinant expression was also evident in embryonated cells of eggs and miracidia as well as the intermediate cellular wall of encysted metacercariae, suggesting its conservation during the course of development of the parasite. Based on the structural relatedness of the cross-reactive fucosyllactose determinant to the antigenic mammalian blood group H trisaccharide, our observations may have implications in serodiagnosis and immunoprophylaxis of schistosomiasis/fascioliasis.

Animals↗
PubMed2003

The immune response of rats to vaccination with the cDNA or protein forms of the cysteine proteinase of Fasciola hepatica.

Our previous experiments have shown that intramuscular injection of Sprague-Dawley rats with a pcDNA 3.1 vector carrying cDNA encoding for a cysteine proteinase (CP) of F. hepatica may induce a high level of protection against subsequent infection with F. hepatica metacercariae (mc). The aim of the present study is to compare the immune response of Sprague-Dawley rats vaccinated intranasally with plasmid containing cDNA of CP of the fluke and intramuscularly or intraperitoneally with the recombinated enzyme protein to challenge with fluke metacercariae. In addition, protection following intranasal DNA vaccination was evaluated. Two experiments were carried out. In the first experiment rats were vaccinated twice with 50microg of cDNA containing plasmid or with 100microg protein of recombinated CP. Three weeks after the second vaccination rats were challenged orally with 25 mc. On days 0, 21, 42 and 63 after the challenge blood samples were collected for the evaluation of white blood cell, eosinophil and specific antibody responses. During the second experiment groups of five male and female rats were vaccinated twice intranasally with CPcDNA then challenged with 30 mc and dissected 5 weeks later. Results obtained in the experiments suggested that intranasal immunisation of rats with CPcDNA seems to favour a Th2 regulated antibody response. Intramuscular or intraperitoneal injections of CP protein stimulate both Th1 and Th2-dependent antibodies. Mean worm burdens found in rats vaccinated intranasally 5 or 10 weeks after the challenge were reduced by 61-75% in comparison with the challenge controls which suggests that intranasal vaccination with CPcDNA may protect hosts against F. hepatica infection.

Animals↗
PubMed1996

Modification of accessory activity of peritoneal cells from Fasciola hepatica infected rats.

The effect of Fasciola hepatica excretory-secretory antigen (ESA) was studied in the modulation of the accessory functions of peritoneal cells (PC) of rats infected with the parasite. PC rats infected with F. hepatica 7 and 14 days previously showed a marked decrease in phagocytic activity against Candida tropicalis (P < 0.007 and P < 0.004, respectively). The same effect was observed when the assay was carried out with PC from animals injected 7 days before with ESA (P < 0.001) including PC previously treated in vitro with ESA. To investigate the effect of ESA on the antigen presenting ability, PC of animals infected before transfer to syngeneic normal rats were stimulated in vitro with either F. hepatica whole antigen (FhWA), ESA or human serum albumin (HSA). Delayed type hypersensitivity (DTH) response to the different antigens studied over a 35 day period was negative in rats transferred with sensitised PC. When these animals were immunised with the corresponding antigen the DTH response became positive. A similar result was obtained in PC receptors from ESA inoculated rats and in vitro stimulated with FhWA or HSA. These data suggest that the alterations observed in the functions of peritoneal cells may, in part be due to the effect of the F. hepatica ESA.

Animals↗
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↗
PubMed1998

Concanavalin A-stimulated proliferation of T cell subset-depleted lymphocyte populations isolated from Fasciola hepatica-infected cattle.

Over 14 weeks, peripheral blood lymphocytes (PBL) were isolated from eight adult cattle which had been orally infected with Fasciola hepatica via trickle infection over a 10-day period. Two age, breed and sex-matched cattle served as controls. CD4+, CD8+ and gammadelta+ T cells were depleted from whole PBL populations by magnetic bead depletion. Lymphocyte proliferation assays demonstrated a transient, but marked elevation in responsiveness to Concanavalin A (Con A) between weeks 2 and 4 post-infection in PBL from infected animals. Proliferative responses to Con A were significantly greater in PBL from infected cattle than uninfected/control cattle over the initial period of the experiment. Con A-stimulated proliferation of PBL isolated from infected cattle followed a similar pattern to PBL responses to F. hepatica antigen. In both whole and subset-depleted lymphocyte populations from infected cattle, proliferative responses to Con A decreased from day 28 post-infection. Depletion of CD4+, CD8+ and gammadelta+ T cell subpopulations significantly augmented responses soon after infection. These findings suggest that the capacity of bovine PBL to proliferate in response to Con A stimulation, was in some way attenuated by F. hepatica infection and proliferative responses due to non-specific activation was suppressed by the coordinated activities of various lymphocyte subsets.

Animals↗
PubMed2003

Cloning, heterologous expression in Escherichia coli and characterization of a protein disulfide isomerase from Fasciola hepatica.

A Fasciola hepatica cDNA clone of 1752 bp was isolated from an adult worm cDNA expression library by immunological screening using a rabbit serum against the excretory-secretory antigens. The nucleotide sequence of the cDNA revealed the presence of an open reading frame of 489 codons which encoded a 55 kDa polypeptide, showing a high degree of homology to protein disulfide isomerases. This putative antioxidant protein cDNA was expressed in Escherichia coli as a GST fusion protein. The cleaved recombinant protein was shown to be biologically active in vitro by mediating the oxidative refolding of reduced RNase. Immunoblotting studies using a specific antiserum raised against the recombinant protein showed the presence of a polypeptide of similar molecular mass in the excretory-secretory extract of the adult parasite. The extracellular location of this protein was also supported by the specific immune responses found against this protein in F. hepatica experimentally infected rabbits.

Amino Acid Sequence↗
PubMed1999

Effects of winter supplementation and antiparasite treatment on the productive performance of milk herd in the central-south region of Chile.

During the winter of 1994, a survey of productive and reproductive parameters was undertaken in the south-central part of Chile (8th Region) on 71 cows provided by four small farms with low production (< 3000 l/year). A low food intake was recorded (8.6 +/- 0.2-12.4 +/- 0.2 kg DM/day) and, as a result, the cows calved with poor body condition score (1.9 +/- 0.04 points on the scale of one-to-five, had milk production between 10.5 +/- 0.4 and 12.8 +/- 0.6 l/day, had a delayed start of ovulatory activity (78.2 +/- 4.6 days) and an interval between calving and conception of 120.3 +/- 4.6 days. In addition, low levels of packed cell volume (PCV) and hemoglobin (HEM) were found, which were considered to be a combined effect of low feed ingestion and parastic infection by Fasciola hepatica. To improve the productive and reproductive parameters, treatments were undertaken in three of the four small farms; a parasiticide treatment was used against the F. hepatica at the beginning of the winter and the availability of forage was increased via the use of supplementary green oats. Spring crops were also cultivated: corn for silage and oats with peas to make hay. The antiparasite treatment and supplementation of forage was associated with better body condition score at calving, improved hematological parameters, increased milk production (17%), reduced number of days to conception (120 vs. 91) and increased conception rate at first service (36 vs. 60%). In the fourth small farm where no interventions were undertaken, the parameters obtained during 1994 remained unchanged.

Animals↗
PubMed2000

Recurrent eosinophilic panniculitis associated with Fasciola hepatica infection.

Eosinophilic panniculitis is characterized by a prominent infiltration of subcutaneous fat with eosinophils. We report a case of Fasciola hepatica infection presenting with eosinophilic panniculitis successfully treated with bithionol. To our knowledge, this is the first report of recurrent eosinophilic panniculitis associated with Fasciola hepatica infection.

Animals↗