1887

Abstract

The conditions for luxuriant axenic cultivation of the trypanosomid flagellate in chemically-defined media were determined. Improvements over the hitherto published media include higher concentrations of amino acids, folic acid (where this is the sole source of pteridine) and purines and the inclusion of threonine and Tween 80. Increased oxygenation was obtained by incubating liquid cultures in tubes in a sloped position. Under these conditions consistently excellent growth (up to 2 × 10 organisms/ml.) was obtained and the useful incubation period could be decreased to 4 days. Using the improved medium, qualitative and quantitative studies on the nutritional requirements of the organism were carried out. Growth failed when any one of the following amino acids was omitted from the basal medium: histidine, phenylalanine, isoleucine, leucine, valine, lysine, arginine, tyrosine, methionine, tryptophan. The omission of threonine caused drastically decreased growth rates and lower yields. It was found that phenylpyruvic acid could replace phenylalanine; cysteine, cystathionine or homocysteine could replace methionine; and citrulline, but not ornithine, could replace arginine for . In addition to the amino acid requirements a nutritional need for the following was demonstrated: a carbohydrate, haemin, a purine, thiamine, riboflavin, pantothenic acid, nicotinic acid or its amide, pyridoxal or pyridoxamine, biotin, folic acid and an unconjugated pteridine (biopterin). The natural purines, their nucleosides and nucleotides are nearly equivalent, on a molar basis, in growth promotion. Numerous substituted purines were also tested for their ability to supply the purine requirement. No exogenous source of pyrimidine for nucleic acid synthesis is necessary provided folic acid is present. Folic acid can be omitted from the growth medium provided thymine and methionine are present. The absolute requirement for biotin could not be demonstrated in the present medium without the use of avidin.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-18-3-621
1958-06-01
2022-01-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/18/3/mic-18-3-621.html?itemId=/content/journal/micro/10.1099/00221287-18-3-621&mimeType=html&fmt=ahah

References

  1. Aaronson S., Nathan H.A. 1954; Utilization of imadazole counterparts of purines in microbial systems. Biochem. Mophys. Acta 15:306
    [Google Scholar]
  2. Broquist H.P., Albrecht A.M. 1955; Pteridines and nutrition of the proto- zoon Crithidia fasciculata . Proc. Soc. exp. Biol., N.Y 89:178
    [Google Scholar]
  3. Cowperthwaite J., Weber M.M., Packer L., Hutner S.H. 1953; Nutrition of Herpetomonas (Strigomonas) culiddarum . Ann. N.Y. Acad. Sci 56:972
    [Google Scholar]
  4. Dewey V.C., Heinrich M.R., Kidder G.W. 1957; Evidence for the absence of the urea cycle in Tetrahymena. J. Protozool 4:211
    [Google Scholar]
  5. Dewey V.C., Parks R.E. Jr Kidder G.W. 1950; Growth responses of Tetrahymena geleii to changes in the basal media. Arch. Biochem 29:281
    [Google Scholar]
  6. Genghof D.S. 1949; The sulfur amino acid requirement of Tetrahymena geleii . Arch. Biochem 23:85
    [Google Scholar]
  7. Gots J.S. 1957; Purine metabolism in bacteria. V. Feed-back inhibition. J. biol. Chem 228:57
    [Google Scholar]
  8. Guthrie R. 1949; Studies of a purine-requiring mutant strain of Escherichia coli . J. Bact 57:39
    [Google Scholar]
  9. Hinton T. 1956; The effect of arginine, ornithine and citrulline on the growth of Drosophila. Arch. Biochem. Biophys 62:78
    [Google Scholar]
  10. Hinton T., Noyes D.T., Ellis J. 1951; Amino acids and growth factors in a chemically defined medium for Drosophila. Physiol. Zool 24:335
    [Google Scholar]
  11. Hutner S.H., Provasoli L. 1955; Comparative biochemistry of flagellates. In Biochemistry and Physiology of Protozoa Hutner S.H., Lwoff A. Ed. by New York:: Academic Press.;
    [Google Scholar]
  12. Johnson W.H., Miller C.A. 1956; A further analysis of the nutrition of Paramecium. J. Protozool 3:221
    [Google Scholar]
  13. Kidder G.W., Dewey V.C. 1948; Dietary factors in the utilization of homocystine. Proc. not. Acad. Sci., Wash 34:81
    [Google Scholar]
  14. Kidder G.W., Dewey V.C. 1949; The biological activity of substituted purines. J. biol. Chem 179:181
    [Google Scholar]
  15. Kidder G.W., Dewey V.C. 1951; The biochemistry of ciliates in pure culture. In Biochemistry and Physiology of Protozoa Lwoff A. Ed. by New York:: Academic Press.;
    [Google Scholar]
  16. Kidder G.W., Dewey V.C. 1955; The purine and pyrimidine requirements of Glaucoma scintillans . Arch. Biochem. Biophys 55:126
    [Google Scholar]
  17. Kidder G.W., Dewey Y.C., Andrews M.B., Kidder R.R. 1949; Tryptophan and nicotinamide in the nutrition of the animal microorganism, Tetrahymena. J. Nutr 37:521
    [Google Scholar]
  18. Kidder G.W., Dewey V.C., Heinrich M.R. 1954; The effect of non-ionic detergents on the growth of Tetrahymena. Exp. Cell. Res 7:256
    [Google Scholar]
  19. Kidder G.W., Dewey V.C., Parks R.E. Jr Heinrich M.R. 1950; Further studies on the purine and pyrimidine metabolism of Tetrahymena. Proc. nat. Acad. Sci., Wash 36:431
    [Google Scholar]
  20. Kidder G.W., Dewey V.C., Parks R.E. Jr Woodside G.L. 1949; Purine metabolism in Tetrahymena and its relation to malignant cells in mice. Science 109:511
    [Google Scholar]
  21. Kidder G.W., Fuller R.C. 1946; The growth response of Tetrahymena geleii W to folic acid and to the Streptococcus lactis R factor. Science 104:160
    [Google Scholar]
  22. Lin S.C., Greenberg D.M. 1954; Enzymatic breakdown of threonine by threonine aldolase. J. gen. Physiol 38:181
    [Google Scholar]
  23. Lwoff M. 1933; Recherches sur la nutrition des trypanosomides. Ann. Inst. Pasteur 51:55
    [Google Scholar]
  24. Miller C.A., Johnson W.H. 1957; A purine and pyrimidine requirement for Paramecium multimicronucleatum . J. Protozool 4:200
    [Google Scholar]
  25. Miller C.A., Van Wagtendonk W.J. 1956; The essential metabolites of a strain of Paramecium aurelia (stock 47–8) and a comparison of the growth rate of different strains of Paramecium aurelia in axenic culture. J. gen. Microbiol 15:280
    [Google Scholar]
  26. Nathan H.A., Cowperthwaite J. 1954; Use of the trypanosomid flagellate, Crithidia fasciculata, for evaluating antimalarials. Proc. Soc. exp. Biol., N.Y 85:117
    [Google Scholar]
  27. Nathan H.A., Cowperthwaite J. 1955; ‘Crithidia Factor’. A new member of the folic acid group of vitamins. J. Protozool 2:37
    [Google Scholar]
  28. Nathan H.A., Hutner S.H., Levin H.L. 1956; Independent requirements for ‘Crithidia Factor’ and folic acid in a trypanosomid flagellate. Nature; Lond.: 178741
    [Google Scholar]
  29. Newton B.A. 1956; A synthetic growth medium for the trypanosomid flagellate Strigomonas (Herpetomonas) oncopetti . Nature; Lond.: 177279
    [Google Scholar]
  30. Nielands J.B. 1952; A crystalline organo-iron pigment from a rust fungus (Ustilago sphaerogena) . J. Amer. ckem. Soc 74:4846
    [Google Scholar]
  31. Nielands J.B. 1953; Biological properties of ferrichrome. Fed. Proc 12:250
    [Google Scholar]
  32. Patterson E.L., Broquist H.P., Albrecht A.M., Von Saltza M.H., Stokstad E.L.R. 1955; A new pteridine in urine required for the growth of the protozoon Crithidia fasciculata . J. Amer. chem. Soc 77:3167
    [Google Scholar]
  33. Pearson W.N. 1949; A purine requiring strain of Photobacterium fisckeri . J. Bact 58:653
    [Google Scholar]
  34. Trager W. 1956; Nutritional requirements of the leptomonads of a hemoflagel-late from lizards (Leishmania tarentolae). J. Protozool.(Suppl.) 3:6
    [Google Scholar]
  35. Van Baalen C., Forrest H.S., Myers J. 1957; Incorporation of radioactive carbon into a pteridine of a blue-green alga. Proc. nat. Acad. Sci., Wash 43:701
    [Google Scholar]
  36. Viscontini M., Loeser E., Karrer P., Hadorn E. 1955; Fluoreszierende Stoffe aus Drosophila melanogaster . Helv. chim. Acta 38:2034
    [Google Scholar]
  37. Wallace F.G. 1943; Flagellate parasites of mosquitoes with special reference to Crithidia fasciculata Léger 1902. J. Parasitol 29:196
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-18-3-621
Loading
/content/journal/micro/10.1099/00221287-18-3-621
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error