Opisthorchis viverrini: Effect of praziquantel on the adult tegument

Abstract

Ultrastructural changes of the tegument of adult liver flukes, Opisthorchis viverrini, after in vitro incubation in Minimal Essential Medium containing 0, 0.1, 1.0 and 10.0 μg/ml of anthelminthic praziquantel for 5, 15, 30, 45 and 60 minutes were investigated by scanning (SEM) and transmission (TEM) electron microscopy. SEM observations showed that the surface damage was composed of blebbing due to the swelling of microvilli, followed later by the disruption of these structures to form lesions that caused the erosion and desquamation of the surface. Sensory papillae, by contrast, appeared relatively unaffected. The surface changes could be observed at all doses but the extent of damage increased with increasing duration of incubation and concentration of the drug. The ventral as well as the dorsal surfaces exhibited similar change, whereas the anterior part tended to be damaged less than the posterior part. Under TEM observations, the earliest sign of changes was the depolymerization of the microtrabecular network in scattered foci, which resulted in the formation of non-membrane-bound vacuoles under microvilli. The basal infoldings also became dilated, and some turned into membrane-bound vacuoles in the basal zone. Subsequently, microvilli became enlarged, and eventually formed blebs that later rupture to form lesion spots as observed in the SEM. Finally, the microtrabecular network in all regions broke down, creating vacuoles of various sizes throughout the tegument, leading to its total disintegration and detachment. The sequence of morphological changes was generally similar at all doses; however, the changes occurred faster at the higher doses and the longer incubation times. In addition, at the longer durations myofilaments in most muscle cells also became depolymerized, while microtubules were unchanged by the drug. Therefore, it is possible that praziquantel, through its induction of Ca2+influx, causes depolymerization of the microtrabecular network that leads to the vacuolization, swelling, blebbing, and eventually the disruption and detachment of the tegument, and the breakdown of myofilaments in the muscle cells.