High performance column liquid chromatographic analysis of selected carboxylic acids in Biomphalaria glabrata patently infected with Schistosoma mansoni

Abstract

High-performance liquid chromatography (HPLC) was used to determine the effects of a patent Schistosoma mansoni infection on certain carboxylic acids in the digestive gland gonad complex (DGG) and hemolymph of Biomphalaria glabrata. An analysis of DGG samples was done using extraction with 50% Locke's solution, cleanup of the extract by anion exchange solid phase extraction (SPE), and ion exclusion HPLC with ultraviolet detection. Hemolymph was applied directly to the SPE column. Acetic, fumaric, malic, and pyruvic acids were detected, confirmed, and quantified at concentrations ranging from 12 to 280 ppm in the DGG and less than 124 to 8,000 microg/dl in the hemolymph. Infection with S. mansoni caused a significant reduction (Student's t test, P< 0.05) in the concentrations of acetic, fumaric, malic, and pyruvic acids in the DGG but not the hemolymph of B. glabrata compared to uninfected cohort snails. The significant reduction of certain carboxylic acids in the DGG of B. glabrata patently infected with S. mansoni suggests that these acids are utilized by the sporocysts and cercariae in the snail tissue, or that the infection stimulates reduced production or increased utilization by the snail tissue.

Advances in the Diagnosis of Human Schistosomiasis

“…Metabolomic analysis of snails following miracidial infection has shown that malic acid and other carboxylic acids are important biomarkers for detecting S. mansoni at different stages of infection in the hemolymph and digestive glands of Biomphalaria alexandrina (330). Changes in lipid levels and concentrations of metabolic by-products, such as pyruvic acid, fumaric acid, and malic acid, in the digestive gland gonad complex of B. glabrata snails infected with S. mansoni have also been identified (331,332). Although the latter approaches are important for understanding the biology of intermediate hosts infected with schistosomes, their application in detecting infection would be limited due to their nonspecificity and the extensive technology required to undertake the analysis.…”

Carboxylic Acids as Biomarkers of Biomphalaria alexandrina Snails Infected with Schistosoma mansoni

“…Also on applying the used technique on the acetic and pyruvic acids, which was not tested previously [20], separation was achieved successfully, and it was possible to separate and quantify the 5 acids at the same time in 1 sample of hemolymph and in 1 sample of tissue extract with lesser retention time than those in other papers [19,22,23]. Without need for solid phase extraction before HPLC analysis, the used technique was a convenient, easier, faster, more accurate, and economical method of carboxylic acid separation.…”

“…The conversions to part per million concentrations (ppm) were done according to the previous report [20].The concentration (ppm) of each acid in snail DGGs was calculated by multiplication of the sample solution concentration interpolated from calibration curve (I, ppm) by the original sample volume (V, ml), and division of the product by the mass of the snail DGG (M, g) and for hemolymph, carboxylic acid concentration (mg/dl) was calculated by multiplying I times V times 100, and division of the product by the hemolymph volume (HV, ml) according to the method of a previous report [19].…”

“…The hemolymph samples were collected from each snail after cleaned with a paper towel by a pasteur pipette inserted through a tiny hole made in the pericardial region then centrifuged at 120 g for 5 min to separate the supernatant from hemocytes and cell debris, then the supernatant was kept at -20℃ till the time of extraction [19].…”

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

“…Both infected and uninfected estivated snails showed significantly decreased maltose and glucose levels compared to the non-estivated infected and uninfected controls. Massa et al (2007) examined the effects of S. mansoni infection on the carboxylic acids of B. glabrata using ion exclusion high performance liquid chromatography (HPLC). They analyzed both the snail DGG and hemolymph.…”