High Performance Thin Layer Chromatographic Analysis of Neutral Lipids in the Feces of BALB/c Mice Infected withEchinostoma caproni

Abstract

High performance thin layer chromatography was used to determine neutral lipid profiles in the feces of BALB/c mice infected with adults of Echinostoma caproni (Trematoda). An approximate 25 worm burden per host induced changes in the neutral lipid profile of the mice at 1 to 5 weeks post infection (PI) relative to the uninfected controls. Infection caused a significant decrease (Student's t-test, P , 0.05) in the triacylglycerol fraction at 3 weeks PI and a significant increase (Student's t-test, P , 0.05) in the free sterol fraction at 2 weeks PI relative to the uninfected controls. Neutral lipid profiles of feces may serve as an indicator of infection by intestinal trematodes in animals and humans.

Metabolic profiling of Echinostoma caproni and Schistosoma mansoni in their definitive and intermediate hosts

“…There have been numerous chromatographic studies on the changes in various metabolites associated with E. caproni infection in both mice and snails. Bandstra et al (2007) used HPTLC to determine the neutral lipid content of mice feces 1-5 weeks postinfection. Samples and standards were chromatographed on either Whatman LHPKDF silica gel plates or EMD Millipore silica gel 60 CF 254 plates developed with the Mangold mobile phase.…”

“…The fecal polar lipids were not useful to analyze as markers of an E. caproni infection since no significant differences were detected between the infected and uninfected samples. Massa et al (2008) extended the study of Bandstra et al (2007) and used the same HPTLC procedures to examine the neutral lipid content of mice feces at 1-7 weeks postinfection. Analysis was done weekly on the fecal samples and consisted of an examination of the free fatty acids, free sterols, triacylglycerols, methyl esters, and steryl esters.…”

“…Vasta et al (2008) used HPTLC to examine the neutral lipid content of the mice urine infected with E. caproni. Folch extracted samples and standards were chromatographed on EMD Millipore silica gel 60 CF 254 plates developed with the Mangold mobile phase; bands were detected using PMA reagent as described in Bandstra et al (2007). To confirm or reject the presence of methyl and steryl esters, the Smith et al (1995) mobile phase consisting of hexanes-petroleum ether-diethyl ether-glacial acetic acid (50:20:5:1), was used on the samples.…”

Studies of echinostomes using chromatography and atomic spectrometry.

“…The classic Folch procedure was used to extract lipids and phospholipids from whole bodies, DGGs, and hemolymph of snails patently infected with Echinostoma caproni (Bandstra et al 2006) ; feces of BALB/c mice infected with E. caproni (Bandstra et al 2007) ; the intestinal mucosa, serum, and liver of ICR mice infected with E. caproni (Albrecht et al 1998 andRivas et al 1998) ; adults of E. caproni , E. trivolvis , and Zygocotyle lunata ; cercariae of E. trivolvis and Echinoparyphium sp. (Muller et al 1999) ; E. trivolvis redial bodies ) ; E. caproni adults and excysted metacercariae (Schneck et al 2004a) ; and SCW and feces from Biomphalaria glabrata infected with E. caproni (Schneck et al 2004b) .…”

“…Similar plates from EMD Chemicals, Inc. (Gibbstown, NJ, an affiliate of Merck KGaA, Darmstadt, Germany) with a preadsorbent zone composed of wide pore silica 50,000 have also been used (Bandstra et al 2007) . The preadsorbent zone adjacent to the main analytical layer facilitates manual sample application of relatively high volumes of biological sample extracts, and the lanes serve as a guide for positioning of the initial zones and the densitometer source slit for scanning chromatograms.…”