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. 1969 Oct;100(1):78–85. doi: 10.1128/jb.100.1.78-85.1969

Enhanced Growth of T-Strain Mycoplasmas with N-2-Hydroxyethylpiperazine-N′-2-Ethanesulfonic Acid Buffer

Richard J Manchee 1, David Taylor-Robinson 1
PMCID: PMC315361  PMID: 5344121

Abstract

The small size of T-strain mycoplasma colonies obtained on agar medium has been a drawback to the study of these organisms. The incorporation of 0.05 mN-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES), a new hydrogen ion buffer, into agar medium at pH 6.8 led to the production of colonies at least 60% greater in diameter than those obtained on media without HEPES at pH 6.5 or 6.0. In addition, the colonies often had the “fried-egg” appearance typical of “classical” large-colony-forming mycoplasmas. The addition of HEPES to liquid medium in the presence of 0.1% urea resulted in a slightly higher number of viable organisms and a corresponding increase in ammonia production. Rapid death of the mycoplasmas occurred on continued incubation of the liquid medium cultures even in the presence of HEPES. The larger colony size facilitated the study of hemadsorption and tissue-culture cell adsorption. Preliminary results of such tests in which these phenomena have been demonstrated are presented. The advantages and disadvantages of having larger colonies are discussed, and the terminology of T-strain mycoplasmas is considered in the light of the present findings.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Black F. T., Rasmussen O. G. Occurrence of T-strains and other mycoplasmata in non-gonococcal urethritis. Br J Vener Dis. 1968 Dec;44(4):324–330. doi: 10.1136/sti.44.4.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CHANOCK R. M., HAYFLICK L., BARILE M. F. Growth on artificial medium of an agent associated with atypical pneumonia and its identification as a PPLO. Proc Natl Acad Sci U S A. 1962 Jan 15;48:41–49. doi: 10.1073/pnas.48.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Conant R. M., Somerson N. L., Senterfit L. B. Immunodiffusion reactions between human sera and Mycoplasma pneumoniae. Proc Soc Exp Biol Med. 1968 Nov;129(2):401–407. doi: 10.3181/00379727-129-33331. [DOI] [PubMed] [Google Scholar]
  4. Del Giudice R. A., Robillard N. F., Carski T. R. Immunofluorescence identification of Mycoplasma on agar by use of incident illumination. J Bacteriol. 1967 Apr;93(4):1205–1209. doi: 10.1128/jb.93.4.1205-1209.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Edward D. G., Freundt E. A., Chanock R. M., Fabricant J., Hayflick L., Lemcke R. M., Rezin S., Somerson N. L., Wittler R. G. Recommendations on nomenclature of the order Mycoplasmatales. Science. 1967 Mar 31;155(3770):1694–1696. doi: 10.1126/science.155.3770.1694. [DOI] [PubMed] [Google Scholar]
  6. Ford D. K. CULTURE OF HUMAN GENITAL "T-STRAIN" PLEUROPNEUMONIA-LIKE ORGANISMS. J Bacteriol. 1962 Nov;84(5):1028–1034. doi: 10.1128/jb.84.5.1028-1034.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ford D. K., MacDonald J. Influence of urea on the growth of T-strain mycoplasmas. J Bacteriol. 1967 May;93(5):1509–1512. doi: 10.1128/jb.93.5.1509-1512.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Good N. E., Winget G. D., Winter W., Connolly T. N., Izawa S., Singh R. M. Hydrogen ion buffers for biological research. Biochemistry. 1966 Feb;5(2):467–477. doi: 10.1021/bi00866a011. [DOI] [PubMed] [Google Scholar]
  9. Hayflick L. Tissue cultures and mycoplasmas. Tex Rep Biol Med. 1965 Jun;23(Suppl):285+–285+. [PubMed] [Google Scholar]
  10. Manchee R. J., Taylor-Robinson D. Haemadsorption and haemagglutination by mycoplasmas. J Gen Microbiol. 1968 Mar;50(3):465–478. doi: 10.1099/00221287-50-3-465. [DOI] [PubMed] [Google Scholar]
  11. Manchee R. J., Taylor-Robinson D. Studies on the nature of receptors involved in attachment of tissue culture cells to mycoplasmas. Br J Exp Pathol. 1969 Feb;50(1):66–75. [PMC free article] [PubMed] [Google Scholar]
  12. Pollack J. D., Somerson N. L., Senterfit L. B. Effect of pH on the immunogenicity of Mycoplasma pneumoniae. J Bacteriol. 1969 Feb;97(2):612–619. doi: 10.1128/jb.97.2.612-619.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Purcell R. H., Taylor-Robinson D., Wong D., Chanock R. M. Color test for the measurement of antibody to T-strain mycoplasmas. J Bacteriol. 1966 Jul;92(1):6–12. doi: 10.1128/jb.92.1.6-12.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SHEPARD M. C., ALEXANDER C. E., Jr, LUNCEFORD C. D., CAMPBELL P. E. POSSIBLE ROLE OF T-STRAIN MYCOPLASMA IN NONGONOCOCCAL URETHRITIS. A SIXTH VENEREAL DISEASE? JAMA. 1964 May 25;188:729–735. doi: 10.1001/jama.1964.03060340027007. [DOI] [PubMed] [Google Scholar]
  15. SHEPARD M. C., LUNCEFORD C. D. EFFECT OF PH ON HUMAN MYCOPLASMA STRAINS. J Bacteriol. 1965 Feb;89:265–270. doi: 10.1128/jb.89.2.265-270.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. SHEPARD M. C. T-Form colonies of pleuropneumonialike organisms. J Bacteriol. 1956 Mar;71(3):362–369. doi: 10.1128/jb.71.3.362-369.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SHEPARD M. C. Visualization and morphology of pleuropneumonialike organisms in clinical material. J Bacteriol. 1957 Feb;73(2):162–171. doi: 10.1128/jb.73.2.162-171.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shepard M. C. Cultivation and properties of T-strains of mycoplasma associated with nongonococcal urethritis. Ann N Y Acad Sci. 1967 Jul 28;143(1):505–514. doi: 10.1111/j.1749-6632.1967.tb27695.x. [DOI] [PubMed] [Google Scholar]
  19. Shepard M. C. Human mycoplasma infections. Health Lab Sci. 1966 Jul;3(3):163–169. [PubMed] [Google Scholar]
  20. Shepard M. C., Lunceford C. D. Occurrence of urease in T strains of Mycoplasma. J Bacteriol. 1967 May;93(5):1513–1520. doi: 10.1128/jb.93.5.1513-1520.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shipman C., Jr Evaluation of 4-(2-hydroxyethyl)-1-piperazineëthanesulfonic acid (HEPES) as a tissue culture buffer. Proc Soc Exp Biol Med. 1969 Jan;130(1):305–310. doi: 10.3181/00379727-130-33543. [DOI] [PubMed] [Google Scholar]
  22. Sobeslavsky O., Chanock R. M. Peroxide formation by mycoplasmas which infect man. Proc Soc Exp Biol Med. 1968 Nov;129(2):531–535. doi: 10.3181/00379727-129-33362. [DOI] [PubMed] [Google Scholar]
  23. Taylor-Robinson D., Addey J. P., Goodwin C. S. Comparison of technics for the isolation of T-strain mycoplasmas. Nature. 1969 Apr 19;222(5190):274–275. doi: 10.1038/222274a0. [DOI] [PubMed] [Google Scholar]
  24. Taylor-Robinson D., Manchee R. J. Novel approach to studying relationships between mycoplasmas and tissue culture cells. Nature. 1967 Dec 30;216(5122):1306–1307. doi: 10.1038/2161306a0. [DOI] [PubMed] [Google Scholar]
  25. Taylor-Robinson D., Purcell R. H. Mycoplasmas of the human urogenital tract and oropharynx and their possible role in disease: a review with some recent observations. Proc R Soc Med. 1966 Nov;59(11 Pt 1):1112–1116. [PMC free article] [PubMed] [Google Scholar]
  26. Williams M. H., Taylor-Robinson D. Aids to the detection of T-strain mycoplasma. Ann N Y Acad Sci. 1967 Jul 28;143(1):394–396. doi: 10.1111/j.1749-6632.1967.tb27682.x. [DOI] [PubMed] [Google Scholar]
  27. Williamson J. D., Cox P. Use of a new buffer in the culture of animal cells. J Gen Virol. 1968 Mar;2(2):309–312. doi: 10.1099/0022-1317-2-2-309. [DOI] [PubMed] [Google Scholar]

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