Effects of tonicity on the release of neutral lipids in Echinostoma caproni adults and observations on lipids in excysted metacercariae.

ABSTRACT

High performance thin layer chromatography was used to analyze neutral lipids in worm incubates isotonic, hypotonic, and hypertonic to the intestinal habitat of adult Echinostoma caproni. Qualitative analysis revealed the presence of free sterols, free fatty acids, triacylglycerols, and a steryl ester/hydrocarbon fraction in all incubate samples. The most abundant neutral lipid fraction released into the incubation medium was the triacylglycerol fraction. This fraction was quantified after worms were maintained for 2 h at 37.5 degrees C in hypertonic (Locke's 2x solution), isotonic (Locke's 0.5x solution) and hypotonic (deionized water) media. Percentages of triacylglycerols on a wet-weight basis found in Locke's 2x, 0.5x, and deionized water were 0.369, 3.23, and 0.242, respectively, suggesting that the optimal medium to obtain maximal excretory-secretory products is the Locke's 0.5x solution. Histochemical staining of whole excysted metacercariae with oil red O did not detect neutral lipids. Analysis of 500 excysted metacercariae incubated for 2 h at 37.5 degrees C revealed that free sterols, free fatty acids, and triacylglycerols were released in amounts of 16.2, 1.59, and 5.34 ng/organism, respectively. Our results were compared with previous studies on neutral lipids in excysted metacercariae and adults of E. trivolvis. Variations in the results of our study compared with others reflect intrinsic differences in the species of echinostome used.