The effects of temperature and salinity on the longevity of Opisthorchis viverrini cercariae: a climate change concern.

Research on the effects of environmental factors influenced by climate change on parasite transmissibility is an area garnering recent attention worldwide. However, there is still a lack of studies on the life cycle of Opisthorchis viverrini, a carcinogenic trematode found in countries of the Lower Mekong subregion of Lao PDR, Cambodia, Myanmar, Vietnam and Thailand. To evaluate the influences of environmental factors water temperature and salinity on the transmissibility of the liver fluke O. viverrini through cercarial stage, longevity of O. viverrini cercaria was examined at different experimental temperatures (22°C, 30°C and 38°C) and salinities (2.5 parts per thousand (PPT), 3.75 PPT and 5 PPT). The results reveal that different temperatures have statistically significant effects on cercarial longevity. The cercariae exhibited a thermostability zone ranging between 22°C and 30°C. Cercarial longevity was significantly shortened when water temperatures reached 38°C. Salinity also plays a key role in cercarial longevity, with cercarial survival significantly shorter at a salinity of 3.75 PPT than at 2.5 PPT and 5 PPT. A combined analysis of salinity and temperature revealed unique trends in cercarial longevity. At all experimental salinities, cercarial longevity was lowest when incubated in 38°C, but statistically significant from cercarial longevity at temperatures of 22°C and 30°C, and salinities of 2.5 PPT and 5 PPT. The results suggest that higher temperatures negatively impact parasite longevity. This reflects that O. viverrini transmission patterns may be impacted by changes in water temperature and salinity resulting from climate change.

Egg-Hatching Mechanism of Human Liver Fluke, Opisthorchis viverrini: A Role For Leucine Aminopeptidases From the Snail Host, Bithynia siamensis goniomphalos.

The human liver fluke Opisthorchis viverrini (Platyhelminthes, Trematoda, Digenea) uses snails of the genus Bithynia as first intermediate host. Peculiarly among trematodes, the eggs of O. viverrini hatch within the digestive tract of its snail host. It remains uncertain whether hatching in this species is mediated through mechanical fracture of the eggshell or by digestion with specific digestive enzymes. This study aimed to characterize enzymes with specific inhibitors and factors involved in the hatching activity of O. viverrini eggs. For measuring egg hatching in vivo, 50 O. viverrini mature eggs were fed to individual Bithynia siamensis goniomphalos snails at various temperature conditions for 24 hr. Ex vivo, mature eggs were incubated with crude snail extract and commercial leucine aminopeptidase (LAP). Egg-hatching of O. viverrini was temperature dependent, with optimal hatching occurring at 24-28 C, with a peak of hatching of 93.54% in vivo and 30.55% ex vivo occurring at these temperatures. Ex vivo hatching rates increased to 45.87% under anaerobic conditions at 28 C. Some 22.70% and 16.21% of heat-killed eggs also hatched within the snail digestive tract and snail extract, respectively, indicating that host molecules are involved in the hatching response. Most eggs hatch in the anterior regions of the digestive tract. Hatching was completely inhibited in the presence of bestatin, an inhibitor of LAP, but not in the presence of phosphatase inhibitors. Bestatin inhibition of hatching was reversible. Finally, egg hatching could be induced by addition of a porcine LAP. The results indicate that this digenean utilizes both LAP of the snail host and movement of miracidia for hatching.

Effects of Bayluscide on Bithynia siamensis goniomphalos, the first intermediate host of the human liver fluke, Opisthorchis viverrini, in laboratory and field trials.

The molluscicidal effects of Bayluscide (niclosamide) were investigated on Bithynia siamensis goniomphalos, the first intermediate host of human liver fluke, Opisthorchis viverrini. Lethal concentrations of 50% (LC(50)) and 95% (LC(95)) against young and adult males were 0.38 and 0.80, 0.42 and 0.86 ppm, respectively. The LC(50) and LC(95) against young and adult females were 0.42 and 0.86, 0.46 and 0.97 ppm, respectively. No significant differences in mortality rate between sexes or snail size (p > 0.05) was detected. Bayluscide-related tissue damage in B. siamensis goniomphalos included detachment of cilia of the epithelial layer of the digestive tract and decreased number of calcium cells. In tests of lethal concentrations of Bayluscide on non-target animals, no lethal effect was observed on Filopaludina martensi martensi (Viviparous snail) but high mortality rates were recorded in Puntius gonionotus fingerling, Ricefish (Oryzias mekongensis) and shrimp (Macrobrachium lanchesteri), but lower in guppy fish (Poecilia reticulata) after 24h exposure. For field trials, sufficient Bayluscide was sprayed in 3 roadside ditches to result in final concentrations of 5, 10 or 20 ppm, with mortality rates on B. siamensis goniomphalos of 10.94, 20.00 and 31.25%, respectively. Non-target snails died in small numbers but no effect was observed in other aquatic vertebrate animals. Field trials of Bayluscide on B. siamensis goniomphalos revealed low mortality rates, suggesting the need for application methods of higher efficacy or that Bayluscide is not suitable for application to operculate snails or snails which are able to escape by burying in mud.