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Lipid class composition of the protozoan Perkinsus marinus, an oyster parasite, and its metabolism of a fluorescent phosphatidylcholine analog

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Lipids

Abstract

Perkinsus marinus is one of two important protozoan parasites of the eastern oyster, Crassostrea virginica. The other is Haplosporidium nelsoni. Lipids extracted from 7-d-old in vitro cultured P. marinus meronts, incubated with fluorescent-labeled phosphatidylcholine (FL PC) and nonincubated P. marinus meronts, were analyzed by a high-performance liquid chromatography (HPLC) system equipped with a diol phase column, in combination with thin-layer chromatography coupled with a flameionization detector (TLC/FID), and high-performance thin-layer chromatography (HPTLC). Various polar and neutral lipid classes were separated by HPLC using a two-gradient solvent system. Five polar lipid classes—phosphatidylcholine (PC), phosphatidylethanolamine (PE), cardiolipin (CL), sphingomyelin (SM), and phosphatidylserine (PS)—were identified from P. marinus extracts. Four neutral lipid classes—triacylglycerol (TAG), steryl ester (SE), cholesterol (CHO), and fatty alcohol—were distinguished. TLC/FID analysis of meront lipids showed that the weight percentages of PC, PE, CL, SM, PS/PI, TAG, SE, and CHO were 21, 10.7, 4, 2.3, 4.3, 48.7, 7.8, and 1.2%, respectively. HPLC and HPTLC analyses revealed the presence of two SM and PS isomers in P. marinus extracts. Perkinsus marinus effectively incorporated FL PC acquired from the medium and metabolized it to various components (i.e., free fatty acid, monoacylglycerol, diacylglycerol, TAG, PE, and CL). Uptake and interconversion of FL PC in P. marinus meronts increased with time. After 48 h the total uptake of fluorescence (FL) was 28.9% of the FL PC added to the medium, and 43% of the incorporated FL resided in TAG.

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Abbreviations

Bodipy:

4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene

CER:

ceramide

CHE:

cholesteryl ester

CHO:

cholesterol

CL:

cardiolipin

DAG:

diacylglycerol

FA:

fatty acid

FAME:

fatty acid methyl ester

FFA:

free fatty acids

FH:

fatty alcohol

FL:

fluorescent-labeled

FTM:

fluid thioglycollate medium

HPLC:

high-performance liquid chromatography

HPTLC:

high-performance thin-layer chromatography

LPC:

lysophosphatidylcholine

MAG:

monoacylglycerol

PC:

phosphatidylcholine

PE:

phosphatidylethanolamine

PG:

phosphatidylglycerol

PI:

phosphatidylinositol

PS:

phosphatidylserine

SE:

steryl ester

SM:

sphingomyelin

TAG:

triacylglycerol

TLC-FID:

thin-layer chromatography coupled with a flame-ionization detector

UV:

ultraviolet

YRW:

York river water

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Author notes

  1. P. Soudant

    Present address: Institut Universitaire Européen de la Mer LEMAR, Université de Bretagne Occidentale, place Nicolas Copernic, 29280, Plouzané, France

Authors and Affiliations

  1. Virginia Institute of Marine Science, College of William and Mary, 23062, Gloucester Point, Virginia

    P. Soudant & F.-L. E. Chu

  2. UMR/CNRS 6521, Université de Bretagne Occidentale, 29285, Brest, France

    Y. Marty

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  1. P. Soudant
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Correspondence to F.-L. E. Chu.

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Soudant, P., Chu, FL.E. & Marty, Y. Lipid class composition of the protozoan Perkinsus marinus, an oyster parasite, and its metabolism of a fluorescent phosphatidylcholine analog. Lipids 35, 1387–1396 (2000). https://doi.org/10.1007/s11745-000-0656-1

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  • DOI: https://doi.org/10.1007/s11745-000-0656-1

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