Isolation and Purification of
            <i>Treponema pallidum</i>
            from Syphilitic Lesions in Rabbits

Hardy, Paul H.; Nell, E. Ellen

doi:10.1128/iai.11.6.1296-1299.1975

Cited by 13 publications

(10 citation statements)

References 12 publications

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“…For unknown reasons all four animals had suboptimal infections at time of harvest, which undoubtedly contributed to the cause of the low treponemal yield. Similar problems have been reported by others (8). 'Protein content was estimated by a dye-binding (4) and the Lowry (17) None detected a Mean relative percentage ± standard error of the mean, n = 3. to 6.2; phosphatidylethanolamine, 0.4 to 0.6; phosphatidylinositol-serine, -0.3; and lysophosphatidylcholine, 1.0.…”

Section: Resultssupporting

confidence: 81%

“…Studies on the lipid composition of T. pallidum could be performed only if organisms were obtained free of rabbit tissue components. To accomplish this, we utilized an extensive purification scheme that included three of the best available tools for obtaining high yields of purified T. pallidum (2,8,25). We obtained 1010 to LIPIDS OF T. PALLIDUM 715 4 x 10's (mean = 2.3 x 1010) treponemes per rabbit, with an average recovery of 45% after purification.…”

Section: Resultsmentioning

confidence: 99%

“…Major problems in research on T. pallidum are the acquisition of sufficient treponemes and their purification from infected tissue. We have obtained high yields of purified organisms for studies of lipid composition by using several well-documented tools: Hypaque gradients (2), Nuclepore membranes (25), and corticosteroidtreated rabbits (8). Baseman et al (2) introduced the use of Hypaque gradients to obtain purified T. pallidum, but heavy bands of tissue material were present in their gradients above the treponemal fractions.…”

Section: Discussionmentioning

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 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: Resultssupporting

confidence: 81%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Unique lipid composition of Treponema pallidum (Nichols virulent strain)

Matthews¹,

Yang²,

Jenkin³

1979

Infect Immun

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“…Although considerable data exist concerning the biology of virulent Treponema pallidum, numerous experimental problems remain. These spirochetes still cannot be readily grown in vitro, and when multiplication is observed treponemal cell densities that are achieved do not even closely approximate values obtained during the direct isolation of T. pallidum from infected rabbit tissue (10,14).…”

mentioning

confidence: 90%

Concanavalin A-mediated affinity film for Treponema pallidum

Baseman¹,

Zachar²,

Hayes³

1980

Infect Immun

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“…The corresponding instantaneous growth rate constant (a = ln 2/tD) is 0.021/h (5). Assuming that protein represents half of the cell dry weight, the dry weight of T. pallidum may be estimated to be 3.8 x 10-13 g (6). During the first 90 min of incubation, 3.8 x 108 treponemes (1.44 x 10-4 g of cells) incorporated 0.2 ,ug of glucose into trichloroacetic acidprecipitable material (Table 1).…”

mentioning

confidence: 99%