Lipids

, Volume 35, Issue 12, pp 1387–1396 | Cite as

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

Article

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.

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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Copyright information

© AOCS Press 2000

Authors and Affiliations

  1. 1.Virginia Institute of Marine ScienceCollege of William and MaryGloucester Point
  2. 2.UMR/CNRS 6521Université de Bretagne OccidentaleBrestFrance

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