Effects of fatty acids on motility retention by Treponema pallidum in vitro

Matthews, Herbert M.; Jenkin, Howard M.; Crilly, Karan S.; Sandok, P L

doi:10.1128/iai.19.3.814-821.1978

Cited by 7 publications

(4 citation statements)

References 31 publications

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“…T. pallidum has also been shown to incorporate "4C-labeled oleic and palmitic acids (25) but was unable to degrade them to 14CO2 (21,25) due to lack of the enzymes of the fl-oxidation pathway (25). The effects of fatty acids on the survival of T. pallidum in vitro have also been found to parallel results obtained in growth studies with cultivable treponemes (9,18).…”

Section: Discussionmentioning

confidence: 78%

“…Some intestinal and oral strains, however, can utilize short-chain fatty acids (26). Recent studies suggest that T. pallidum may also have lesions in lipid metabolism similar to many cultivable treponemes (9,18,21,25).Lipids comprise 15 to 20% of the cellular dry weight of the cultivable treponemes (10, 16). The major component is a glycolipid, monoglycosyldiglyceride (MGDG), containing galactose generally as the sugar moiety.…”

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confidence: 99%

“…Fatty acids and fatty acid methyl ester standards were obtained from The Hormel Institute, Austin, Minn., and Nu-Chek Prep, Inc., Elysian, Minn.; neutral lipid and phospholipid standards were from Supelco, Inc., Bellefonte, Pa., and Avanti Biochemicals, Inc., Birmingham, Ala.; and Silica Gel H was from E. Merck AG, Darmstadt, Germany. A dyebinding protein assay (4) kit was purchased from Bio-Rad Laboratories, Richmond, Calif. All solvents were distilled before lipid analyses (28 Purification of T. pallidunL Treponemes were extracted aerobically from testes of corticosteroidtreated rabbits 11 to 14 days postinfection as previously described (18) until 35 to 50 ml of suspension was obtained from each testis. Serum-free Eagle medium containing 4 mg of fatty acid-free bovine serum albumin (Miles Laboratories, Inc., Elkhart, Ind.)…”

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Unique lipid composition of Treponema pallidum (Nichols virulent strain)

Matthews¹,

Yang²,

Jenkin³

1979

Infect Immun

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The lipid composition of Treponema pallidum (Nichols virulent strain) was determined after purification of the organisms from the infected testes of corticosteroid-treated rabbits by differential centrifugation, filtration through Nuclepore membranes, and sedimentation in Hypaque density gradients. The total lipids were comprised of 32.2% neutral lipids, mainly cholesterol, and 67.8% phospholipids consisting of phosphatidylcholine (32.1%), sphingomyelin (14.8%), cardiolipin (13.0%), phosphatidylethanolamine (6.2%), phosphatidylinositol-serine (1.2%), and lysophosphatidylcholine (0.4%). Monoglycosyldiglyceride, a glycolipid comprising 25 to 50% of the total lipid of all Treponema previously examined, was not detected. The fatty acid composition was similar but quantitatively distinct from that of the infected testes tissue.Attempts to cultivate Treponema pallidum, the causative agent of syphilis, in vitro have failed since its discovery almost 75 years ago. Largely due to this, little is known about the organism's lipid metabolism or composition. Many cultivable Treponema require long-chain fatty acids for growth (9,16,22) since they can neither synthesize, 8-oxidize, nor desaturate fatty acids (1,10,19). Some intestinal and oral strains, however, can utilize short-chain fatty acids (26). Recent studies suggest that T. pallidum may also have lesions in lipid metabolism similar to many cultivable treponemes (9,18,21,25).Lipids comprise 15 to 20% of the cellular dry weight of the cultivable treponemes (10, 16). The major component is a glycolipid, monoglycosyldiglyceride (MGDG), containing galactose generally as the sugar moiety. MGDG comprises 25 to 50% of the total lipid of each of the approximately 20 treponemal strains of genital, oral, and intestinal origin so far examined (10,16,19) including T. hyodysenteriae (our unpublished data), the only cultivable pathogenic Treponema (12). Although Vaczi et al. (29) investigated the lipid composition of T. pallidum (Budapest strain), as well as four cultivable treponemes, they reported no quantitative data about the individual lipids or gave any indication of the glycolipid content of the organisms.Since T. pallidum cannot be cultivated in vitro, the organism is obtained for laboratory studies from the testes of experimentally infected rabbits. A major impedance of research on T. pallidum is the difficulty of obtaining large yields of the organism free of rabbit testicular material. In recent years, however, the use of corticosteroid-treated rabbits (8), Nuclepore filters (25), and Hypaque gradients (2) has proven remarkably useful in reducing this problem. We have used all of these tools to obtain high numbers of purified T. pallidum for lipid analysis. Infected rabbit testicular tissue and T. phagedenis biotype reiterii were also examined as comparative controls. Our results have shown that T. pallidum appears to be unique among Treponema in that the major lipid component of other Treponema, namely MGDG, was not detected in this human pathogen.MATERIALS AND METHODS C...

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Section: Discussionmentioning

confidence: 78%

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confidence: 99%

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confidence: 99%

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Unique lipid composition of Treponema pallidum (Nichols virulent strain)

Matthews¹,

Yang²,

Jenkin³

1979

Infect Immun

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“…The metabolism of rapidly degenerating T. pallidum may be different than that of a treponemal population exhibiting better stability in the rate of isotope incorporation and in the number of motile microorganisms available during incubation. We had developed a serum-free medium in which up to 90% of the treponemes remained motile for 96 h (12). Our system of incubation held some promise to facilitate future studies on substrate utilization and turnover by T. pallidum.…”

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Radiolabeling of Treponema pallidum (Nichols virulent strain) in vitro with precursors for protein and RNA biosynthesis

Sandok¹,

Jenkin²

1978

Infect Immun

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We observed uptake of [U-'4C]serine, U-'4C-labeled amino acid hydrolysates, and [2-'4C]uracil by virulent Treponema pallidum in vitro for at least 96 h. No uptake of [2-'4C]thymine, [1-'4C]pyruvate, [U-14C]pyruvate, and [2-'4C]uridine was detected. Treponemal protein and RNA biosynthetic activity was identified by erythromycin inhibition of amino acid and uracil uptake. Radioactivity due to uptake of radiolabeled amino acids by residual testicular cells in the cultures remained at background levels regardless of the presence or absence of cycloheximide. Accumulation of the radiolabeled substrates by T. pallidum proceeded at a linear rate for 48 to 96 h during incubation in vitro. The longevity of substrate uptake using the system of incubation described will facilitate future studies on the metabolism of the microorganism to help determine essential growth factors and environmental conditions for multiplication of T. pallidum in vitro. Until recently, the most commonly accepted techniques to study the physiology of Treponema pallidum in vitro were dependent upon the use of light microscopy for direct counts and on the exploitation of the rabbit to evaluate retention of virulence by the microorganism (6, 26). Now, it is becoming increasingly apparent that these techniques are not completely satisfactory for physiological studies on T. pallidum in vitro. For example, Baseman and Hayes observed inhibition of radiolabeled amino acid uptake by erythromycin in cultures of T. pallidum

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