Parasitology Research

, Volume 76, Issue 8, pp 643–647 | Cite as

A new medium containing antibiotics for the xenic cultivation ofEntamoeba gingivalis

  • John T. Gannon
  • Harald A. B. Linke
Original Investigations

Abstract

Diamond's trypticase-yeast extract-serum-gastric mucin (TYSGM-9) medium was studied for its suitability to support the xenic growth of the oral protozoanEntamoeba gingivalis. Amoebic growth was found to be best when the inoculum for transfer was 0.1 ml, the incubation temperature was 35°C, and the interval between transfers was 48 h. These parameters were also useful for controlling the growth in xenic stock cultures, In addition, bacterial growth in xenic stock cultures, which had a direct effect on the activity ofE. gingivalis trophozoites, was kept to a minimum by addition of the antibiotics piperacillin, erythromycin, neomycin, and penicillin. Varying substitutions and selected supplementations of TYSGM-9 medium led to the development of an improved medium forE. gingivalis. Supplements most beneficial for the growth ofE. gingivalis trophozoites were ascorbic acid, ferric ammonium citrate, and special NCTC 107 vitamin mixture. As compared with TYSGM-9 medium (6–10E. gingivalis trophozoites observed per field), the newE. gingivalis medium supported excellent growth (16–20E. gingivalis trophozoites observed per field during optimal growth) of the oral protozoan. The medium is suitable for clinical isolation ofE. gingivalis.

Keywords

Ascorbic Acid Penicillin Erythromycin Bacterial Growth Clinical Isolation 

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References

  1. Boeck WC, Drobohlav J (1925) The cultivation ofEntamoeba histolytica. Am J Hyg 5:371–407Google Scholar
  2. Clayton JP, Ball GH (1954) Effects of penicillin onEntamoeba gingivalis in cultures with bacteria from the human mouth. J Parasitol 40:347–352Google Scholar
  3. Cleveland LR, Collier J (1930) Various improvements in the cultivation ofEntamoeba histolytica. Am J Hyg 12:606–613Google Scholar
  4. Diamond LS (1968) Improved method for the monoxenic cultivation ofEntamoeba histolytica Schaudinn, 1903 andE. hystolytica-like amebae with trypanosomatids. J Parasitol 54:715–719Google Scholar
  5. Diamond LS (1982) A new liquid medium for the xenic cultivation ofEntamoeba histolytica and other lumen dewelling protozoa. J Parasitol 68 (5):958–959Google Scholar
  6. Diamond LS (1983) Lumen dwelling protozoa:Entamoeba, trichomonads, andGiardia In: Jensen JB (ed) In-vitro cultivation of protozoan parasites. CRC Press, Boca Raton, Florida, pp 65–109Google Scholar
  7. Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation ofEntamoeba histolytica and otherEntamoeba. Trans R Soc Trop Med Hyg 72 (4):431–432Google Scholar
  8. Gannon JT (1987) Physiological growth studies ofEntamoeba gingivalis. Ph.D. Thesis, New York University (published by University Microfilms International, Ann Harbot, Michigan)Google Scholar
  9. Gannon JT, Linke HAB (1989a) Studies on the microflora associated with xenic cultures ofEntamoeba gingivalis. Microbios 58: 95–100Google Scholar
  10. Gannon JT, Linke HAB (1989b) Growth studies on xenic cultures ofEntamoeba gingivalis using established media. Int J Parasitol 19: 835–838Google Scholar
  11. Griffin AM, McCarten WG (1949) Sterols and fatty acids in the nutrition of entozoic amoebae in cultures. Proc Soc Exp Biol Med 72:645–648Google Scholar
  12. Gros G (1849) Fragments d'helminthologie et de physiologie microscopique. Bull Soc Imp Nat Moscou 22 (2):549–573Google Scholar
  13. Howitt BF (1925) The cultivation ofEntamoeba gingivalis (Gros). Univ Calif Publ Zool 28:66–122Google Scholar
  14. Linke HAB, Gannon JT, Obin JN (1989) Clinical survey ofEntamoeba gingivalis by multiple sampling in patients with advanced periodontal disease. Int J Parasitol 19:803–808Google Scholar
  15. Michaelson JB, DeLamater JN, Talpis L (1953) Comparison of starches used in the culture ofEntamoeba histolytica. Am J Trop Med Hyg 2:219–226Google Scholar
  16. Morrison RT, Boyd RN (1975) Organic chemistry, 3rd edn. Allyn and Bacon Inc., Boston, p 1129Google Scholar
  17. Nisizawa K, Pigman W (1959) The composition and properties of the mucin clot from cattle submaxillary glands. Arch Oral Biol 1:161–170Google Scholar
  18. Reeves RE, Meleney HE, Frye WM (1957) A modified Shaffer-Frye technique for the cultivation ofEntamoeba histolytica and some observations on its carbohydrate requirements. Am J Hyg 66:56–62Google Scholar
  19. Spingarn CL, Edelman MH (1950) The effect of rice powder on the growth of cultures ofEntamoeba histolytica. Am J Trop Med 30:629–634Google Scholar
  20. Van Vliet HHDM, Opdenkamp JAF, Van Deenan LLM (1975) Lipid auxotrophy and the effect on lipid composition ofEntamoeba invadens. J Protozool 22 (3):428–432Google Scholar
  21. Wantland WW, Wantland EM, Winquist DL (1963) Collection, identification and cultivation of oral protozoa. J Dent Res 42:1234–1241Google Scholar
  22. Whistler RL, Bemiller JN, Paschall EF (1984) Starch: chemistry and technology, 2nd edn. Academic Press, New York, pp 107–108Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • John T. Gannon
    • 1
  • Harald A. B. Linke
    • 1
  1. 1.Department of MicrobiologyNew York University Dental CenterNew YorkUSA

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