The energy metabolism of the adult common liver fluke,Fasciola hepatica

In adultFasciola hepatica glucose is broken down to volatile fatty acids, carbon dioxide and lactate. In the cytosol glucose is converted into malate and lactate and malate is metabolized further in the mitochondrial compartment via a dismutation. The dismutation of malate can proceed under anaerobic conditions. To a limited extent oxygen can function as terminal electron acceptor.It is shown that the reaction catalysed by fumarate reductase is a reversible reaction. The direction of the reaction depends on the relative concentrations of fumarate and succinate.Mechanisms of ATP production and electron transport under aerobic and anaerobic conditions are discussed. It is concluded that the energy metabolism of the adult liver fluke is adapted to the life in an environment with a low oxygen content.

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Kurzfassung Beim adultenFasciola hepatica wird Glucose in flüchtige Fettsäuren, Kohlendioxid und Laktate gespalten. Im Zytosol wird die Glucose in Malate und Laktate gespalten und Malate werden im Mitochondrienbereich durch Redox-Dismutation weiter verstoffwechselt. Die Dismutation der Malate kann unter anaeroben Bedingungen ablaufen. In gewissem Ausmass kann Sauerstoff terminal Elektronen binden.Es wird gezeigt, dass die Reaktion, die durch Fumaratreduktase katalysiert wird, reversibel ist. Der Ablauf der Reaktion hängt von den relativen Konzentrationen der Fumarate und Succinate ab.Ferner werden die Mechanismen der ATP-Produktion und des Elektronentransports unter aeroben und anaeroben Bedingungen diskutiert. Zusammenfassend wird festgestellt, dass der Energiestoffwechsel des adulten grossen Leberegels an ein Leben in einer sauerstoffarmen Umwelt adaptiert ist.

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Resume Chez les douves adultes le glucose est dégradé en acides gras volatil, gaz carbonique et lactate. Dans le cytosol le glucose est convertit en malate et en lactate et le malate est métabolisé ensuite dans la mitochondrie selon un phénomène de transformation biochimique. La transformation biochimique du malate peut se réaliser dans des conditions anaérobies. Dans une certaine mesure l'oxygène peut fonctionner comme un accepteur terminal d'électrons.Il est démontré que la réaction catalysée par la fumarate réductase est réversible. Le sens de la réaction dépend de la concentration relative en fumarate et succinate.Le mécanisme de production de l'ATP, du transport des électrons dans des conditions aérobies et anaérobies est discuté. Il est retenu que le métabolisme énergétique de la douve adulte est adapté à la vie dans un environnement pauvre en oxygène.

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Riassunto Negli adulti diFasciola hepatica il glucosio si scinde in acidi grassi volatili, biossido di carbonio e lattato. Nel citosol il glucosio è convertito in malato e lattato, ed il malato è ulteriormente metabolizzato nel comparto mitocondriale attraverso un processo di dismutazione. La dismutazione del malato può proseguire in condizioni di anaerobiosi. In misura limitata l'ossigeno può agire come accettore terminale di elettroni.E'stato dimostrato che la reazione catalizzata dalla fumarato-reduttasi è una reazione reversibile. La direzione della reazione dipende dalle concentrazioni relative di fumarato e di succinato.Vengono discussi i meccanismi di produzione di ATP e del trasporto di elettroni in condizione di aerobiosi e di anaerobiosi. Si conclude che il metabolismo energetico dell'adulto diFasciola hepatica è adattato alla vita in un ambiente con bassi livelli di ossigenazione.

     

Liver fluke,Fasciola hepatica in New Zealand

Free-living members of the Mustelidae and feral cats (Felis catus) inhabiting farmland in the southern half of the North Island of New Zealand were examined for evidence of leptospiral infection. Nine stoats (Mustela erminea), 9 ferrets (Putorious putorious) and 4 weasels (Mustela nivalis) showed no serological or bacteriological evidence of infection. No leptospires were isolated from the kidneys of 11 feral cats but 2 animals had low titres to pomona and ballum respectively. All animals examined were from ecosystems where ballum infection was endemic in rodents; thus predator-chain transmission does not appear to be an important natural route of transmission for this serovar.     

Impact of compound alpha treatment in vivo on egg production by the liver fluke, Fasciola hepatica

Sheep infected with the triclabendazole-susceptible Cullompton isolate of Fasciola hepatica were treated with compound alpha at a dosage of 15 mg/kg at 12 weeks post-infection. Adult flukes were recovered from the bile ducts at 24h, 48 h and 72 h post-treatment (pt). They were processed for whole mount analysis, histology and transmission electron microscopy of the female reproductive system: specifically, the uterus, Mehlis' gland, ovary and vitellaria. As judged by the appearance of the uterus, normal egg production ceased within 24h of treatment; this phenomenon preceded significant changes to the other reproductive organs. Over the 3-day pt period, there was a progressive decline in the number of oogonia in the ovary, together with an increase in the number of eosinophilic and apoptotic oocytes and vacuolation and shrinkage of the ovarian tubules. There was a shift in the cell population within the vitelline follicles at 48 h pt, with relatively greater numbers of mature vitelline cells and fewer immature cells. The follicles were vacuolated and the shell globule clusters in the mature cells were disorganised. Greater disruption was seen at 72 h pt, with a reduction in the size of the follicles and rupture of cells, releasing their content into the lumen of the follicles. These histological observations were confirmed and extended at the TEM level. Thus, examination of electron micrographs showed that disruption of the shell globule clusters was evident at 48 h pt, which coincided with the start of the breakdown of the mature cells and of the nurse cell network. These degenerative changes were more conspicuous at 72 h pt. In the Mehlis' gland, shrinkage and vacuolation of the cells and their cytoplasmic extensions became progressively greater from 48 h to 72 h pt, and secretory activity declined. The changes in the reproductive organs and inhibition of egg production are put in context of the overall time-course of drug action.     

The effect of albendazole and triclabendazole on colchicine binding in the liver fluke Fasciola hepatica

Albendazole (ABZ) and its sulfoxide (SX) and sulfone (SO) metabolites inhibit the binding of 3H-colchicine, a ligand with high affinity for tubulin to homogenate preparations of the liver fluke Fasciola hepatica. The relative potency of these compounds is SX greater than ABZ greater than SO. The benzimidazoles (cambendazole, parbendazole, oxibendazole and mebendazole), when tested at a concentration of 10 microM, also inhibited colchicine binding to fluke homogenates. However, a potent new benzimidazole flukacide, triclabendazole (TCB), was without effect on colchicine binding to F. hepatica homogenates. When intact flukes were exposed in vitro to 10(-5)M SX for as little as 5 min the subsequent binding of 3H-colchicine to fluke homogenates was significantly reduced. However, flukes recovered from sheep either 12 or 24 h after treatment with ABZ did not have a decreased ability to bind colchicine, although the non-specific binding was higher in flukes from treated sheep, suggesting some interaction of drug with tubulin in vivo. ABZ, SX and SO were effective in preventing embryonation of fluke eggs at doses as low as 0.01 microM, but TCB was without effect at concentrations as high as 10 microM. The results suggest that ABZ exerts at least part of its anthelmintic effect by interaction with fluke tubulin.     

Distribution and habitats of the snail Lymnaea truncatula, intermediate host of the liver fluke Fasciola hepatica, in South Africa

This paper focuses on the geographical distribution and habitats of Lymnaea truncatula, the intermediate, snail host of the liver fluke, Fasciola hepatica, as reflected by the 723 samples in the database of the National Freshwater Snail Collection, Potchefstroom, South Africa. The 221 different loci (1/16-degree squares) on record reflect an extensive but discontinuous distribution, except in Lesotho and in parts of the Mpumalanga, Gauteng and North West provinces of South Africa. Although recorded from 12 different types of waterbody, it was mostly (42.0%) recovered from swamps. Most samples (45.8%) were collected in habitats with slow-flowing water. A muddy substratum was recorded for 62.5% of the samples. Most samples (86.3%) were collected in habitats with a mean annual air temperature of 10-20 degrees C, and more than 69% came from localities with a mean annual rainfall of 600-900 mm. An integrated decision tree constructed from the data indicated that temperature and types of waterbody play a decisive role in determining the presence of L. truncatula in a given area. A temperature index calculated for all mollusc species ranked L. truncatula second in a total of 53 species according to its association with low temperatures. It remains to be established whether its distribution is indeed discontinuous, and whether its preference for a particular habitat, amphibious habits and ability to aestivate could have resulted in some populations having been overlooked during surveys.     

Galectin secretion and binding to adult Fasciola hepatica during chronic liver fluke infection of sheep

Galectins are increasingly recognised as important mediators of immune homeostasis and disease regulation, but comparatively little is known about their role in parasite infection. This study investigates the interaction between two ovine galectins, galectin-11 and galectin-14, and the parasitic liver fluke, F. hepatica. Galectin-14 was found in eosinophils infiltrating the tissue surrounding infected bile ducts and secreted in the connective tissue, while galectin-11 was specifically induced in epithelial cells of bile ducts from infected sheep. Strong nuclear staining was observed for galectin-11. Both galectins were found to be secreted into the bile fluid of parasite infected sheep, and were also detected in the excretory/secretory products of adult flukes, following their removal from the ovine host. Recombinant galectin-14, but not recombinant galectin-11, was found to bind specifically to the surface tegument of adult flukes in a carbohydrate dependent manner. This study shows for the first time that both galectin-14 and galectin-11 are produced in liver tissue after chronic liver fluke infection and that they can directly interact with the parasite in the bile ducts. Galectin-11 may also be involved in epithelial cell turnover and cancerogenesis.     

Cathepsin L proteinases as vaccines against infection with Fasciola hepatica (liver fluke) in ruminants

Fasciolosis, caused by trematodes of the genus Fasciola, is one of the most important diseases of farmed ruminants in temperate and tropical zones. The appearance of Fasciola hepatica populations that are resistant to common flukicidal drugs means that new methods of treatment will soon be required. The future prospect for the development of anti-liver fluke vaccines is optimistic and given their consumer acceptability and environmental friendliness, offer the best way forward. Cathepsin L proteases (Fhe CL 1 and Fhe CL 2), secreted by liver flukes at all stages of their development in the mammalian host, are believed to play important roles in facilitating parasite migration (tissue degradation), feeding and immuno-evasion. The authors consider them prime targets for which new vaccines can be developed. Vaccine studies in cattle and sheep have shown that protection levels of up to 72 and 79 per cent, respectively, can be obtained with immunisations of cathepsin Ls in Freunds' adjuvant. The vaccine also exhibited high anti-embryonation/anti-fecundity effects on parasites that survived in vaccinated animals and thus could have a major impact on the transmission of disease to the intermediate host. While natural infections in sheep and cattle appear to elicit non-protective Th2 immune responses, the authors' studies indicate that the protection induced by vaccination involves elements of a Th1 response.     

Innate and adaptive resistance of Indonesian Thin Tail sheep to liver fluke: a comparative analysis of Fasciola gigantica and Fasciola hepatica infection

In the current study, three independent trials directly compared Fasciola gigantica and Fasciola hepatica infection of ITT sheep. In all trials, F. hepatica infection resulted in higher worm burden recoveries and greater physiological damage to ITT sheep. Developmental differences of the two Fasciola species were also observed during the first twelve weeks of a primary infection, where the migration and growth of F. hepatica was more rapid than F. gigantica. Various immunological blood parameters were measured and indicated similar kinetics in the humoral and cellular responses during the time course of infection with each Fasciola species. In contrast to F. hepatica infection, we demonstrate an innate and adaptive comparative ability of ITT sheep to resist the early stages of infection with F. gigantica infection. Unraveling the mechanisms leading to this differential resistance may potentially lead to new methods for the control of fasciolosis and other human liver flukes.     

Evaluation of the comparative efficacy of a moxidectin plus triclabendazole pour-on solution against adult and immature liver fluke, Fasciola hepatica, in cattle

The objective of this study was to evaluate the efficacy of a pour-on solution containing moxidectin plus triclabendazole (MOX plus TCBZ) against immature and adult stages of the liver fluke in cattle and compare the efficacy with other commercially available preparations. To this end, 104 male Holstein-Friesian calves aged between 3 and 4 months, were randomly allocated to 13 groups of eight animals each, and infected with approximately 500 Fasciola hepatica metacercariae. One group remained untreated, four groups were treated with MOX plus TCBZ at a dose rate of 0.1mL/kg, four other groups were treated with ivermectin (IVM) plus clorsulon injectable at a dose rate of 0.02mL/kg, and the remaining four groups were treated with IVM plus closantel pour-on at a dose rate of 0.1mL/kg. Each treatment was applied to one of the groups at 4 weeks, 6 weeks, 8 weeks and 12 weeks after the experimental infection. At necropsy (99-102 days after infection), all untreated animals were infected with a minimum of 30 flukes. The MOX plus TCBZ treated animals had significantly (P<0.0001) lower fluke counts compared to the untreated control animals at all time points after treatment. Efficacy against 8-week old and adult flukes was >99.5%. For 6-week old immature fluke, the efficacy was 98.0% and for 4-week old immature fluke the efficacy was 90.9%. The IVM plus closantel pour-on treated animals had significantly lower fluke counts compared to the untreated control animals for adult and 8-week old flukes (P<0.0001), and for 6-week old flukes (P=0.002). The efficacy was 26.8%, 68.2%, 90.6% and 99.3% against 4-week, 6-week and 8-week old immature flukes, and adult flukes respectively. The IVM plus clorsulon treated animals had significantly lower fluke counts compared to the untreated control animals for adult (P<0.0001) and 8-week old (P<0.05) flukes. The efficacy was 29.7%, 43.4%, 53.2% and 99.2% against 4-week, 6-week and 8-week old immature flukes, and adult flukes respectively. For treatments at 4, 6 and 8 weeks after infection, the fluke counts were significantly (P<0.0001) lower for the MOX plus TCBZ treatment than for IVM plus closantel or IVM plus clorsulon. The results confirm the high efficacy (>90%) of the MOX plus TCBZ pour-on combination against 4-week old to adult liver fluke in cattle. The IVM plus closantel pour-on combination was effective (>90%) against 8-week old and adult flukes, but had low efficacy against 4- and 6-week old fluke. The IVM plus clorsulon injectable combination was effective (>90%) against adult fluke only.     

Performance characteristics of an enzyme-linked immunosorbent assay for the detection of liver fluke (Fasciola hepatica) infection in sheep and cattle

AIM: To validate an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against liver fluke (Fasciola hepatica) in sheep and cattle sera. METHODS: Gold-standard sera from sheep and cattle of known infection status, i.e. sera from non-infected animals and from animals known to be infected with F. hepatica were assayed with a commercially available ELISA and results analysed by ROC analysis. RESULTS: The ROC analysis suggested cut-offs that were considerably lower than those suggested by the manufacturer, yet the ELISA performed with high sensitivity and specificity, 98 to 100%, respectively for sheep and cattle sera. For bovine sera, particularly good discrimination between positive and negative sera was observed. Infection in experimentally infested animals could be demonstrated 7-8 weeks earlier than with classical parasitological techniques. CONCLUSIONS: The analysis of the ELISA's performance demonstrated high sensitivity and specificity. ROC analyses optimised the cut-off point suggested by the manufacturer of the commercial diagnostic assay. Diagnosis of infection with F. hepatica was achieved much earlier than is possible with current parasitological techniques. This could help with the control of fasciolosis, enabling treatment before clinical manifestation of the disease.     

Performance characteristics of an enzyme-linked immunosorbent assay performed in milk for the detection of liver fluke (Fasciola hepatica) infection in cattle

AIM: To determine the performance characteristics of an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against liver fluke (Fasciola hepatica) in bovine milk. METHODS: Serum and milk from liver fluke infected and non-infected cattle was assayed in a commercially available enzyme-linked immunosorbent assay. Serum test results were used to determine the "gold standard" infection status of cattle and milk ELISA results assessed by ROC analysis. RESULTS: ROC analysis suggested changes to the ELISA protocol, arriving at milk dilutions assayed considerably higher than those suggested by the manufacturer. With those changes, the ELISA performed with high sensitivity and specificity, 95 and 98.2%, respectively, for individual bovine milks (relative to sera). For bovine tank milks, sensitivity was lower, with bulk milks only testing positive if 60% or more of cattle milking in the herd were infected. CONCLUSIONS: The analysis of the ELISA's performance when used on individual bovine milks demonstrated high sensitivity and specificity. ROC analyses optimised the assay conditions and cut-off point suggested by the manufacturer for this commercial diagnostic assay. This would help with the identification and control of fasciolosis, enabling simpler sample collection.     

Liver fluke (Fasciola hepatica) in slaughtered sheep and cattle in New Zealand, 1984-85

A total of 17175 lambs livers and 9322 sheep livers randomly selected from animals slaughtered at export meatworks in 1984 were examined for liver fluke infection. Futher sheep livers from some regions were examined in 1985. The origins of the animals examined were recorded and used to estimate regional prevalences of infection. Infection in lambs was first detected in February and the prevalence increased after then; the mean prevalence in July was 2.7% in the North Island and 2.0% in the South Island . In adult sheep in 1984, regional prevalences in the North Island ranged up to 16.9% (>10% in South Auckland, East Coast, Hawkes Bay and Taranaki) and in the South Island up to 29.4% in Westland (18% in Nelson). The overall prevalence in the North Island was 7.5% and in the South Island 1.1%, about double that recorded in 1969. Data for 1985 were incomplete but prevalences tended to be higher than in 1984. From April 1984 to August 1985 the origins of all lines of cattle with fluke-affected livers were recorded in all abattoirs and export meatworks. During 1985, 16 147 infected lines were traced to all countries in the North Island, with over 60% from the Whangarei, Hamilton, Taumarunui, Gisborne and Hastings areas. In the South Island , over 90% of 1585 infected lines originated from Westland, Waimea, Grey, Inangahua and Buller Counties.