Ultrastructural changes to the tegumental system and gastrodermal cells of adult Fasciola hepatica following treatment in vivo with a commercial preparation of myrrh (Mirazid).

An in vivo study in the laboratory rat model has been carried out to monitor changes to the tegument and gut of adult Fasciola hepatica following treatment with myrrh ('Mirazid'). Rats infected with the triclabendazole-resistant Dutch isolate were dosed orally with Mirazid at a concentration of 250 mg/kg and flukes recovered 2, 3 and 7 days post-treatment (pt). The flukes were processed for examination by scanning and transmission electron microscopy. A variety of changes to the external surface were observed, culminating in the sloughing of the tegumental syncytium. Internal changes to the syncytium and tegumental cell bodies were more severe and were evident from 2 days pt onwards. Swelling of the basal infolds (leading to flooding of the surface layer) and a decline in secretory body production were the major changes seen. The gastrodermal cells were less severely affected than the tegument, pointing to a trans-tegumental route of uptake for Mirazid by the fluke. Some loss of muscle fibres in the main somatic muscle layers was observed, which may be correlated with the decline in movement of flukes seen at recovery.

Fasciola hepatica: a comparative survey of adult fluke resistance to triclabendazole, nitroxynil and closantel on selected upland and lowland sheep farms in Northern Ireland using faecal egg counting, coproantigen ELISA testing and fluke histology.

In order to investigate the incidence and distribution of adult fluke resistance to the fasciolicide tricalbendazole (TCBZ) amongst populations of Fasciola hepatica in sheep flocks in Northern Ireland (NI), individual rectal faeces samples were collected from 3 groups of 20 sheep, before (pre-dose), and 21 days after (post-dose) treatment of the animals with TCBZ, nitroxynil or closantel, on each of 13 well-managed sheep farms distributed across the province. The efficacy of each flukicide was determined for each farm, using faecal egg count reduction (FECRT) and F. hepatica coproantigen ELISA testing. In certain flocks, 2 sheep with high pre-dose faecal egg counts (FEC) were killed 3 days and 21 days respectively after TCBZ treatment, and the histology of the fluke reproductive organs was compared with that of flukes from untreated sheep, and from sheep treated with nitroxynil or closantel 2 days prior to death, using haematoxylin and eosin (H&E) staining and an in situ hybridisation method (TdT-mediated dUDP nick end labelling [TUNEL]) to demonstrate apoptosis. Results from FECRT revealed that in all flocks with a high fluke burden, TCBZ was ineffective in treating chronic fasciolosis, and this finding was generally supported by the results of the coproantigen reduction test (CRT). The histology of reproductive organs of flukes from TCBZ-treated sheep in these flocks was normal, when compared with untreated flukes, and this, together with the FECRT and CRT findings, indicated a likely diagnosis of TCBZ resistance in all the flocks with a high fluke burden. In contrast, nitroxynil and closantel were found to be fully effective against TCBZ-resistant flukes in each of the flocks bearing a high chronic fluke burden. All of the flocks with a high fluke burden and TCBZ resistance were managed on lowland in the South and East of NI. Upland flocks, in the North and West, had low fluke burdens, or were clear of infection; and FECs were too low to allow valid resistance testing. The study highlights the high level of penetration of TCBZ resistance throughout F. hepatica populations in areas of intensively managed sheep production with a high level of fluke challenge. Further, it emphasises the importance of pre-emptive chemotherapeutic action against chronic fasciolosis, using flukicides effective against the egg-producing adult flukes to minimise pasture contamination for the next season's lamb crop. This study also exemplifies the use of several complementary methods (FECRT; CRT; fluke histology; comparative anthelmintic efficacy testing) for confirmation of a diagnosis of fluke drug resistance.

Triclabendazole progress report, 2005-2009: an advancement of learning?

Triclabendazole (TCBZ) remains the drug of choice for treating infections of the liver fluke, Fasciola hepatica in livestock and has become the main drug used to treat human cases of the disease as well. Cases of resistance in livestock continue to be reported, suggesting that the problem is increasing. In order to address the problem, there is a need for better understanding of drug action. A 'state-of-play' review on different aspects of TCBZ activity was published by the present author in 2005. The main purpose of the current review is to assess what progress has been made in the past four years towards understanding the main aspects of drug activity, including drug pharmacokinetics and pharmacodynamics and an understanding of the mechanism(s) of resistance. Also, what advances have been made in identifying alternative compounds and using drug combinations to enhance TCBZ activity. Stemming from a number of in vivo studies, it has become evident that fluke isolates of differing sensitivity to TCBZ differ in some of their biological parameters, and information on this interesting phenomenon will be presented. An update on the use of TCBZ for human fascioliasis is also given. The review will indicate what progress has been made, but will also highlight areas that remain inadequately understood and require greater research focus.

The structure, composition, formation and possible functions of calcareous corpuscles in Trilocularia acanthiaevulgaris Olsson 1867 (Cestoda, Tetraphyllidea).

Mature calcareous corpuscles in the juvenile (stomach) worms of Trilocularia acanthiaevulgaris comprise a number of concentric lamellae interspersed with areas of flocculent material. Each lamella is composed of a pair of membranous rings to which amorphous, non-crystalline material is attached. The process of corpuscle formation is intracellular, beginning with the autophagic break-down of the cytoplasm to produce a central vacuole within a parenchymal cell. The vacuole enlarges until only a thin layer of cytoplasm remains at the periphery and the nucleus is displaced to one end of the cell. Paired, concentrically arranged membranes are laid down beneath the peripheral cytoplasmic layer and eventually occlude the central vacuole. X-ray analysis of the corpuscles indicates the presence of calcium, phosphorus, sulphur, zinc and molybdenum, with the major peaks representing calcium, phosphorus and sulphur. Calcium appears to be bound to the lamellae rather than associated with the material between lamellae. The possible functions of the corpuscles are discussed in relation to the biology of T. acanthiaevulgaris and its developmental sequence in the dogfish gut.