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PubMed1983

Fasciola hepatica: inhibition of phosphoenolpyruvate carboxykinase, and end-product formation by quinolinic acid and 3-mercaptopicolinic acid.

Quinolinic acid and 3-mercaptopicolinic acid act as inhibitors of Fasciola hepatica phosphoenolpyruvate carboxykinase. Low concentrations of these compounds (0.1 mM quinolinate and 0.01 mM 3-mercaptopicolinate) resulted in noncompetitive inhibition, which became mixed inhibition at higher concentrations (1.5 and 0.15 mM, respectively). 3-mercaptopicolinic acid proved to be a much more potent effector than quinolinic acid. Both quinolinic acid and 3-mercaptopicolinic acid caused a significant reduction in the total amount of end product excreted, again 3-mercaptopicolinate being more effective than quinolinate. When glucose was present in the medium, both propionate and acetate levels fell significantly with both inhibitors; however, only 3-mercaptopicolinic acid caused an effect in the absence of glucose.

Acetates↗
PubMed1995

Evaluation of nitroxynil and closantel activity using ELISA and egg counts against Fasciola hepatica in experimentally and naturally infected cattle.

The responses of cattle infected with Fasciola hepatica to treatment with nitroxynil or closantel were monitored by faecal egg counts and by ELISA assay of anti-fluke antibodies. A first trial with experimentally infected heifers showed an increase in anti-fluke antibody titre as early as 2 weeks post-infection. Eggs were first detected in the faeces 10 weeks after infection. Egg output increased steadily over the next 8 weeks and then rapidly decreased. Treatment of a 20-week infection with nitroxynil was followed by a slow decrease in antibody titre 4 weeks later. This decrease continued over the next 40 weeks, but returned to pre-infection levels in only 2 out of 4 animals. The faecal egg count fell to zero 2 weeks after treatment and remained so for the following 30 weeks, although 1 animal produced a few eggs 32 and 34 weeks post-treatment. Within this period, neither diagnostic technique discriminated between this persistently infected animal and the others. In a second trial, 45 cattle from a naturally infected herd were treated with nitroxynil or closantel. The faecal egg counts of the treated cattle were zero within the following 2 months, whereas there were eggs in the faeces of the control (untreated) group. Nevertheless, the treated cattle showed a small, non-significant drop in anti-fluke antibody titre. These results demonstrate the need for new tools to monitor and evaluate accurately the efficacy of anthelmintic treatment.

Animals↗
PubMed2004

Fasciola hepatica: the growth and larval productivity of redial generations in Galba truncatula subjected to miracidia differing in their mammalian origin.

Experimental infections of Galba truncatula with 4 isolates of Fasciola hepatica miracidia differing by their mammalian origin (cattle, nutrias, rabbits, or sheep) were carried out to determine if parasite origin had an effect on the number of free rediae, their growth, and their larval productivity in each redia category. The mammalian origin of miracidia had a significant influence on the numbers of free rediae (they were higher in cattle-group snails) and the lengths of rediae (they were lower in rabbit groups). The redia category had also a significant effect on body and pharyngeal measurements. In all groups, the majority of cercariae (55.8-63.2%) were produced by the daughter rediae (R2a rediae) originating from the first mother redia. Compared with the other groups, the mean number of cercariae at day 49 postexposure was twice as high in cattle groups. In contrast, the mean number of daughter rediae produced by each second-appearing mother redia or each R2a redia was higher in the nutria, rabbit, and sheep groups. The mammalian origin of F. hepatica miracidia had an effect on the number of live rediae, their length, and their redial and cercarial productivity.

Animals↗
PubMed1976

The influence of temperature on the infectivity of Fasciola hepatica miracidia to Lymnaea truncatula.

Experiments were performed to study the effect of water temperature on the host-finding capacity (snail localization, attachment, and penetration) of Fasciola hepatica miracidia. Specimens of Lymnaea truncatula were exposed to miracidia labeled in vivo with radioselenium, and the radioactivity which subsequently was confined to the snails was taken as a measure of the host-finding capacity of the parasite. The minimum temperature required for host-finding was 5 to 6 C and the optimum temperature was in the range between 15 and 26 C. The lack of host-finding capacity below 5 to 6 C could be reversed experimentally by raising the temperature. A clear inverse relationship was demonstrated between the environnd 24 C the host-finding capacity ceased after 24 to 30, 20 to 24, and 13 to 20 hr, respectively.

Animals↗
PubMed1995

The effect of triclabendazole ("Fasinex") on protein synthesis by the liver fluke, Fasciola hepatica.

The effect of the active sulphoxide metabolite of the anthelmintic triclabendazole (TCBZ-SX, 15-50 micrograms ml-1) on the incorporation of radioactively labelled [14C] leucine by adult Fasciola hepatica tissue slices was measured by liquid scintillation counting. In addition, the ability of the microfilament-disrupting drug, cytochalasin B, and the microtubule-disrupting drug, tubulozole-C, to inhibit protein synthesis, was assessed by similar methods and compared with TCBZ-SX. The established protein synthesis inhibitors, cycloheximide and actinomycin D were used as positive controls. All the drugs showed a significant inhibition of protein synthesis, albeit to different extents; however, TCBZ-SX was the most potent, with no significant difference between its effect and that of cycloheximide or actinomycin D. Moreover, the concentration of TCBZ-SX, above 15 micrograms ml-1, had little further influence on incorporation of [14C] leucine. This investigation demonstrates the inhibitory effect of TCBZ-SX, cytochalasin B and tubulozole-C on protein synthesis in F. hepatica and confirms the qualitative observations made in several previous ultrastructural studies.

Animals↗
PubMed1987

The anthelmintic efficacy of netobimin against experimental infections of Fasciola hepatica in sheep.

Netobimin (coded SCH 32481, Schering Corporation), a new broad-spectrum anthelmintic having both fasciolicidal and nematocidal properties was evaluated for efficacy against mature Fasciola hepatica infections in sheep. The trial was conducted with 30 cross-bred spring lambs, each experimentally infected with 250 F. hepatica metacercariae. A single treatment of netobimin was administered at 17 weeks post-infection (PI) by oral drench at 7.5 or 20 mg kg-1 body weight while 10 animals remained as untreated controls. At necropsy (either 1 or 2 weeks post-treatment), the mean number of adult flukes recovered from the control, 7.5 and 20 mg kg-1 groups were 94.7, 35.9 and 8.8, respectively. The resulting efficacies were 62% (P less than or equal to 0.05) and 90.7% (P less than or equal to 0.01), respectively. No clinical signs of fascioliasis were noted in any sheep during the trial. No signs of toxicosis nor any adverse reactions to the drug were observed.

Animals↗
PubMed2003

The parasitic trematode Fasciola hepatica exhibits mammalian-type glycolipids as well as Gal(beta1-6)Gal-terminating glycolipids that account for cestode serological cross-reactivity.

Neutral glycosphingolipids from sheep-derived Fasciola hepatica liver flukes were isolated and characterized both structurally and serologically. After HPLC fractionation, glycolipids were analyzed by linkage analysis, enzymatic cleavage, and MALDI-TOF as well as electrospray ionization mass spectrometry. Obtained results revealed the presence of two types of neutral glycolipids. The first group represented mammalian-type species comprising globo- and isoglobotriaosylceramides (Gal(alpha1-4)Gal(beta1-4)Glc(1-1)ceramide and Gal(alpha1-3)Gal(beta1-4)Glc(1-1)ceramide, respectively) as well as Forssman antigen (GalNAc(alpha1-3)GalNAc(beta1-3/4)Gal(alpha1-4/3)Gal(beta1-4)Glc(1-1)ceramide). Applying Helix pomatia agglutinin, recognizing terminal alpha-linked GalNAc, to cryosections of adult flukes, the latter glycolipid could be localized to the F. hepatica gut. As Forssman antigen from the parasite and sheep host led to identical MALDI-TOF MS profiles, this glycolipid might be acquired from the definitive host. As a second group, highly antigenic glycolipids were structurally characterized as Gal(beta1-6)Gal(beta1-4)Glc(1-1)ceramide, Gal(beta1-6)Gal(alpha1-3/4)Gal(beta1-4)Glc(1-1)ceramide and Gal(beta1-6)Gal(beta1-6)Gal(alpha1-3/4)Gal(beta1-4)Glc(1-1)ceramide, the latter two structures of which exhibited both isoglobo- or globo-series core structures. Terminal Gal(beta1-6)Gal1-motifs have previously been shown to represent antigenic epitopes of neogala-series glycosphingolipids from tape worms. Using human Echinococcus granulosus infection sera, Gal(beta1-6)Gal-terminating glycolipids could be allocated to the gut in adult liver fluke cryosections. Corresponding neogala-reactive antibodies in F. hepatica infection serum were detected by their binding to E. granulosus and Taenia crassiceps neogala-glycosphingolipids. These antibodies might contribute to the known serological cross-reactivity between F. hepatica and parasitic cestode infections.

Animals↗
PubMed1988

Comparative efficacy of triclabendazole, nitroxynil and rafoxanide against immature and mature Fasciola hepatica in naturally infected cattle.

In two trials the fasciolicidal activities of triclabendazole, nitroxynil and rafoxanide were assessed in cattle naturally infected with predominantly immature stages of Fasciola hepatica. Tablets containing 900 mg triclabendazole were administered orally at a dose rate of 12 mg/kg bodyweight. Rafoxanide and nitroxynil were used at a dose rate of 10 mg/kg, rafoxanide being given orally and nitroxynil by subcutaneous injection. Based on faecal egg counts nine weeks after treatment the efficacies were calculated to be 100 per cent for triclabendazole and 95.0 per cent for nitroxynil in the first trial and 98.4 per cent for triclabendazole and 52.9 per cent for rafoxanide 15 weeks after treatment in the second trial. In the first trial five animals from each of the three groups were slaughtered and their fluke burdens counted. Compared with the untreated control group the reductions in the fluke burdens were 96.9 per cent in triclabendazole treated cattle and 76.4 per cent in the nitroxynil treated group.

Animals↗
PubMed1994

The effect of the sulphoxide metabolite of triclabendazole ('Fasinex') on the tegument of mature and immature stages of the liver fluke, Fasciola hepatica.

The effects of the novel benzimidazole, triclabendazole (TCBZ) ('Fasinex', Ciba-Geigy), in its active sulphoxide metabolite form (TCBZ-SX), on the tegumental ultrastructure of Fasciola hepatica were determined in vitro by transmission electron microscopy (TEM), using both intact flukes and tissue-slice material. At a concentration of 15 micrograms/ml, the tegument of the whole adult fluke showed ultrastructural changes only after prolonged time-periods, with vacuolation at the base of the syncytium and accumulation of T2 secretory bodies in the tegumental cells. At a concentration of 50 micrograms/ml, with both whole flukes and tissue-slices, the tegument appeared extremely abnormal with accumulation of secretory bodies towards the base of the syncytium. With longer incubation times, the tegument was completely sloughed away and the tegumental cells became synthetically inactive. The tegument of the 3-week-old juvenile became progressively convoluted at the apex, while in the basal regions there was severe vacuolation. In the tegumental cells, there were accumulations of T1 secretory bodies. These results confirm TCBZ as a potent fasciolicide, being very effective in disrupting the fluke tegument. They may go some way to explain the mode of action of this important fasciolicide.

Animals↗
PubMed2005

A retrospective study on the metacercarial production of Fasciola hepatica from experimentally infected Galba truncatula in central France.

A retrospective study of the experiments performed during the past 15 years on infections of Galba truncatula with Fasciola hepatica was carried out to determine what susceptible populations of snails might be used for the commercial production of metacercariae, and to examine this metacercarial production in relation to the characteristics of snail infections. Of the four groups of snail populations studied, the ablest snails to sustain a complete larval development of F. hepatica originated from populations living on siliceous soils at 600 m and more in altitude. In contrast, snail populations living along river banks on siliceous soils were inappropriate due to the poor characteristics of snail infections (high snail mortality, low prevalence of snail infections, and low number of cercariae produced). Except for these last populations, 86-87% of cercaria-shedding (CS) snails in the other populations shed less than 300 cercariae, even if a maximum of 1,772 cercariae were obtained from a single snail. The date of the first cercarial shedding at 20 degrees C began during week 7 or 8 PE for 80.1-83.5% of CS snails. Most metacercariae (82.0-85.9% of the total production) were recorded during the first 10 days of the patent period. In these conditions, the authors collected metacercarial production up to the beginning of week 10 PE (20 degrees C) and did not use snails that shed their cercariae during the following weeks due to too low numbers of parasites. This method allows to have a continuous production of metacercariae over time by using successive groups of infected snails, each being separated from the other by a fortnight's time.

Animals↗
PubMed1979

Fasciola hepatica: attempts to induce protection against infection in rats and mice by injection of excretory/secretory products of immature worms.

In vitro excretory/secretory products of 4-week (immature) and 8-week-old (mature) Fasciola hepatica parasites, derived from rats, were injected together with adjuvant into naive rats and mice. Resistance to infection was assessed in rats by counting adults in the bile ducts at 9 weeks, or in mice by recording deaths after oral challenge with a high dose of viable metacercariae. Exposure of rats to excretory/secretory products of immature F. hepatica conferred a significant degree of resistance which was comparable to the level of resistance induced following oral administration of a low number of metacercariae. No protection against infection was seen in rats injected with excretory/secretory products from mature, bile duct-derived worms. In mice, no obvious mouse strain variation in susceptibility to first infection existed and hypothymic nude mice were as susceptible to infection as intact mice. As determined by protection against death, vaccination with excretory/secretory products derived from immature F. hepatica was without effect in mice. It is concluded that "host protective antigens", at least for rats, were present in the excretory/secretory products of immature F. hepatica larvae.

Animals↗
PubMed1998

Characterization of Fasciola hepatica redial generations by morphometry and chaetotaxy under experimental conditions.

Morphometric and chaetotactic studies were carried out on the body and cephalic regions of the rediae of Fasciola hepatica (Trematoda: Fasciolidae) in order to precisely identify the different redial generations of this trematode in Lymnaea truncatula under experimental infection. At day 49 post-exposure at 20 degreesC, the length of the redia was significantly higher in the first group of the first generation (R1a) compared with successive generations, R1b, R2a and R2b/R3a. The width of the body was similar in the R1a, R1b, and R2a rediae, but was significantly lower in the R2b/R3a groups. The intrapharyngeal cavity of R1a rediae was significantly wider compared with the R1b, R2a, and R2b/R3a groups, whereas the pharyngeal wall was significantly thicker in the R2b/R3a rediae compared with the R1b and R2a groups. Four other measurements, namely the maximum length and width of the pharynx, diameter of the mouth, and width of intestine, also showed significant variations in relation to pharyngeal morphology and age of infection. Discriminant analysis based on these measurements demonstrated that 98% of the rediae were readily categorized into the four groups identified. The number of perioral sensillae ranged from 126 to 160 but a significant difference was only noted between the mean values of the first generation and those of the group R2b/R3a. From these parameters, the maximum width of the pharyngeal lumen was found to be the best characteristic in the identification of the redial generations.

Animals↗
PubMed1990

The effect of the hydrogen ionophore closantel upon the pharmacology and ultrastructure of the adult liver fluke Fasciola hepatica.

The present study describes the effects of the H+ ionophore and anthelmintic closantel upon the in vitro motility and in vivo ultrastructure of the liver fluke, Fasciola hepatica. At a concentration of 50 micrograms/ml, closantel caused an initial stimulation, then suppression of activity, which was accompanied by an increase in muscle tone and led to a spastic paralysis within 2 h. The pattern of response was similar at lower concentrations, although the initial stimulation was not always evident, but the onset of paralysis could be reached more quickly. Scanning electron microscopy revealed gross surface damage from 24 h onwards in vivo, in the form of erosion of the anterior and posterior extremities of the fluke and large-scale sloughing of the tegument on both dorsal and ventral surfaces. Tegumental changes prior to sloughing included some swelling of the basal infolds and an apical accumulation of T1 secretory bodies. In the underlying tegumental cells there was reduced secretory activity and the mitochondria were consistently swollen and deformed. Reduced secretory activity was a feature of the gastrodermal cells as well; these cells were characterized by swollen, electron-lucent mitochondria, vesiculated GER cisternae and apical blebbing of packets of cytoplasm. The vitelline follicles became severely disrupted as a result of the breakdown of the nurse cell cytoplasm. The stem and intermediate type 1 (It1) cells rounded up and showed nuclear abnormalities. There did not appear to be a severe disruption of shell protein production in the intermediate vitelline cells, but there was a noticeable absence of glycogen in the mature vitelline cells. The effects of closantel are discussed in relation to its proposed mode of action as an uncoupler of oxidative phosphorylation.

Animals↗
PubMedDate not supplied

Phenotype of hepatic infiltrates and hepatic lymph nodes of lambs primarily and challenge infected with Fasciola hepatica, with and without triclabendazole treatment.

The phenotype of the hepatic inflammatory infiltrate and hepatic lymph nodes (HLN) was analysed in lambs primarily and challenge infected with Fasciola hepatica. Group 1 was primarily and challenge infected with two doses of 200 metacercariae (mc) each and was non-treated. Trickle infection was administered to five groups: group 2 was challenge infected and non-treated; group 3 was primarily infected and non-treated; group 4 was primarily infected and treated with triclabendazole (TCBZ) at 12 weeks postinfection (wpi); group 5 was treated at 4 wpi and challenge infected and group 6 was treated at 12 wpi and challenge infected. An uninfected group was used as the control. The distribution of T cell subpopulations (CD3, CD4 and CD8), and B cells (CD79alpha, IgM, IgG) was analysed. The hepatic inflammatory infiltrate was represented mainly by CD3 and CD4 T cells, and B cells (CD79alpha, IgG). These infiltrates were more severe (P < 0.05) in primarily (group 3) or challenge (groups 2, 5 and 6) trickle infected lambs than in the group single challenge infected (group 1). Cellular changes in HLN consisted in an increase of CD4 over CD8 T cells and an increase of B cells and IgG+ plasma cells, and they were more severe in primarily and challenge trickle infected groups than in the group infected with two larger doses of mc, although significant differences were not found with respect to all challenge trickle infected groups. The strong local cellular and humoral immune responses did not protect against subsequent infections, neither in non-treated lambs (group 2) nor in lambs treated with TCBZ at 4 wpi (group 5) or 12 wpi (group 6).

Animals↗
PubMed1987

Reevaluation of the potential of Fasciola hepatica antigens for immunization against Schistosoma mansoni infection.

Previous workers have reported that significant resistance to Schistosoma mansoni cercarial challenge can be induced in mice by immunization with various Fasciola hepatica antigen preparations. We have attempted to reproduce these observations and have also carried out similar experiments in rats. In our hands, however, CBA mice did not develop either antibodies against the schistosomular surface, or significant resistance to challenge after immunization with heterologous antigens. Similar results were obtained in Fischer rats, but rats did develop significant resistance after immunization with homologous (schistosomular) antigens, even without adjuvant, and the homologous 'protective' antigens could be separated by antibody-affinity chromatography using serum from rats vaccinated with irradiated cercariae. Thus, we found no evidence for the existence of shared 'protective' antigens between S. mansoni and F. hepatica, and suggest that this is further evidence that where heterologous resistance can be demonstrated between these two parasites, it is probably due to immunologically non-specific mechanisms.

Adjuvants, Immunologic↗
PubMed2002

Pathological and immunohistochemical study of the liver and hepatic lymph nodes of sheep chronically reinfected with Fasciola hepatica, with or without triclabendazole treatment.

Hepatic lesions and changes in the hepatic lymph nodes were examined in groups of sheep infected, and sometimes reinfected, with metacercariae of Fasciola hepatica, with or without treatment with triclabendazole (TCBZ). Hepatic damage was more severe in sheep given small repeated ("trickle") infections than in those given two larger, spaced doses. Inflammatory infiltrates of eosinophils, CD3+ T cells, CD79 alpha+ B cells and IgG+ plasma cells, and the presence of granulomata, were particularly marked in trickle-infected groups, suggesting that the host response to certain fluke or egg antigens causes severe hepatic damage. TCBZ administration induced the elimination of flukes and healing of the majority of hepatic lesions but did not prevent severe hepatic damage produced by later infections.

Animals↗
PubMedDate not supplied

Fasciola hepatica: characterization of somatic and excretory-secretory antigens of adult flukes recognized by infected sheep.

The immune response of sheep to somatic components and excretory/secretory products of adult Fasciola hepatica, was studied during an experimental infection. Antibodies against adult fluke somatic and excretory/secretory antigens were detected by thin layer immunoassay from the second week post infection. Similarly, the results of Western blot analysis showed specific recognition of several components as early as two weeks after infection. However, an increase in the number and intensity of bands with time of infection was observed in the patterns of antigenic recognition. Most notorious was the strong reactivity of all infected sheep sera towards components of 20-23 kDa in the somatic preparation and components of 23-27 kDa in the excretory/secretory products of adult F. hepatica, specially noticeable after the sixth week post-infection. Since these polypeptides were recognized by all infected animals, they could play an important role in the diagnosis of sheep fascioliasis.

Animals↗