Alterations in the phospholipid composition and morphology of ovine erythrocytes after intravenous inoculation of Corynebacterium pseudotuberculosis

Kim A. Brogden From the National Animal Disease Center, Agricultural Research Service, USDA (Brogden), and Department of Veterinary Physiology and Pharmacology, Iowa State University (Engen), Ames, IA 50011.

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Richard L. Engen From the National Animal Disease Center, Agricultural Research Service, USDA (Brogden), and Department of Veterinary Physiology and Pharmacology, Iowa State University (Engen), Ames, IA 50011.

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Summary

Corynebacterium pseudotuberculosis produces a sphingomyelin-specific phospholipase D exotoxin that is a major determinant in the pathogenesis of caseous lymphadenitis. The effect of this exotoxin on erythrocytes was assessed during experimentally induced infection of sheep. Blood was drawn at timed intervals, and the phospholipid composition of erythrocytes was determined by use of high-performance liquid chromatographic analysis of membrane extracts. Erythrocyte morphology was determined by use of transmission electron microscopy. Significant (P ≤ 0.05) decreases in erythrocyte membrane sphingomyelin content and significant (P ≤ 0.05) increases in phosphatidylglycerol content were observed 30 minutes after iv inoculation of C pseudotuberculosis. The concentration of other phospholipids remained unchanged. Initially, spherostomatocytes were formed that later became pitted at the cell surface. These pits or invaginations appeared as numerous vacuoles at the periphery of thin-sectioned cells. Pitting became progressively worse, leading to an extensive scalloped cell surface. Alterations in the phospholipid composition and morphology of ovine erythrocytes may contribute to pathophysiologic findings in sheep with acute infection induced by C pseudotuberculosis.

Summary

Corynebacterium pseudotuberculosis produces a sphingomyelin-specific phospholipase D exotoxin that is a major determinant in the pathogenesis of caseous lymphadenitis. The effect of this exotoxin on erythrocytes was assessed during experimentally induced infection of sheep. Blood was drawn at timed intervals, and the phospholipid composition of erythrocytes was determined by use of high-performance liquid chromatographic analysis of membrane extracts. Erythrocyte morphology was determined by use of transmission electron microscopy. Significant (P ≤ 0.05) decreases in erythrocyte membrane sphingomyelin content and significant (P ≤ 0.05) increases in phosphatidylglycerol content were observed 30 minutes after iv inoculation of C pseudotuberculosis. The concentration of other phospholipids remained unchanged. Initially, spherostomatocytes were formed that later became pitted at the cell surface. These pits or invaginations appeared as numerous vacuoles at the periphery of thin-sectioned cells. Pitting became progressively worse, leading to an extensive scalloped cell surface. Alterations in the phospholipid composition and morphology of ovine erythrocytes may contribute to pathophysiologic findings in sheep with acute infection induced by C pseudotuberculosis.

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