Effects of Echinostoma caproni miracidia dose on the neutral and polar lipids of Biomphalaria glabrata as determined by high-performance thin-layer chromatography
Treść / Zawartość
The effects of a 5 versus 25 miracidia exposure of Echinostoma caproni on the lipid composition of Biomphalaria glabrata was studied using high performance thin layer chromatography (HPTLC)-densitometry. A 50 miracidia dose was not used because such a high level of exposure caused severe snail mortality by 3 weeks post-exposure (PE). Lipids were determined in the digestive-gland gonad complex (DGG) of the exposed snails and in the uninfected matched controls at 2 and 4 weeks PE. Extraction of lipids from DGGs was carried out by the Folch method with chloroform-methanol (2:1), and extracts were analyzed on Analtech HPTLC-HLF pre-adsorbent silica gel plates with measurement of separated bands using a CAMAG Scanner 3. For neutral lipids the mobile phase was petroleum ether-diethyl ether-glacial acetic acid (80:20:1) and the detection reagent was 5% ethanolic phosphoric acid, and for polar lipids chloroform-methanol-deionized water (65:25:4) mobile phase and 10% cupric sulfate in 8% phosphoric acid detection reagent were used. No significant differences in the concentrations of free sterols, free fatty acids, triacylglycerols, phosphatidylcholine, and phosphatidylethanolamine were seen at 2 weeks PE in any of the groups. At 4 weeks PE, the free fatty acid concentration increased significantly in the snails exposed to 25 miracidia compared to that of the 5 miracidia/snail group or the controls. Elevation of the free fatty acid fraction in the high dose snail group suggested that some changes occurred in the lipid metabolism of the snails in that group as a function of miracidia dose.
- Beideman D., Fried B., Sherma J. 2013. Effects of coexposure with Echinostoma caproni and Schistosoma mansoni miracidia on neutral and polar lipids of Biomphalaria glabrata as determined by high-performance thin-layer chromatography-densitometry and observations on snail survival and fecundity. Journal of Liquid Chromatography & Related Technologies, 36, 2489–2496. DOI: 10.1080/10826076.2013.790772.
- Hunsberger A., Beideman D., Fried B. 2012. Effects of various Echinostoma caproni (Trematoda) miracidia doses on the survival and fecundity of Biomphalaria glabrata snails. Newsletter of the American Malacological Society 43, 6–7.
- Mangold, H. K. 1969. Aliphatic lipids. In: Thin Layer Chromatography, 2nd Ed., Stahl E., Ed. Springer-Verlag: New York, NY; pp 363–421.
- Tunholi-Alves V.M., Lustrino D., Tunholi-Alves V.M., de Mello-Silva C.C., Maldonado A. Jr., Rodrigues Mde L, Pinheiro J. 2011a. Changes in the reproductive biology of Biomphalaria glabrata infected with different doses of Echinostoma paraensei miracidia. Journal of Invertebrate Pathology, 106, 192–195. DOI: 10.1016/j.jip.2010.09.006.
- Tunholi-Alves V.M., Tunholi V.M., Golo P., Lustrino D., Maldonado A. Jr., Bittencourt V.R., Rodrigues Mde L., Pinheiro J. 2011b. Lipid levels in Biomphalaria glabrata infected with different doses of Echinostoma paraensei miracidia. Experimental Parasitology, 128, 212–216. DOI: 10.1016/j.exppara.2011.03.009.
- Ulmer M.J. 1970. Notes on rearing snails in the laboratory. In Experiments and Techniques in Parasitology; MacInis, A.J., Voge, M., Eds.; W. H. Freeman and Co.: San Francisco, CA; pp. 143–144.
- Wagner H., Horhammer L., Wolff P. 1961. Thin layer chromatography of phosphatides and glycolipids. Biochemische Zeitschrift, 334, 175–184.