Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1983 Nov;42(2):501–509. doi: 10.1128/iai.42.2.501-509.1983

Production of cholera toxin-like toxin by Vibrio mimicus and non-O1 Vibrio cholerae: batch culture conditions for optimum yields and isolation of hypertoxigenic lincomycin-resistant mutants.

W M Spira, P J Fedorka-Cray
PMCID: PMC264457  PMID: 6642640

Abstract

Vibrio mimicus 61892, isolated in 1977 from a case of watery diarrhea in Bangladesh, produces an enterotoxin which possesses activity in Y-1 mouse adrenal cells and in rabbit ileal loops which is identical to the prototype cholera toxin (CT) produced by Vibrio cholerae 569B. The neutralization of the adrenal cell activity of 61892 toxin and 569B CT by homologous and heterologous antisera generates parallel titration curves which show complete neutralization in all cases. Paired titrations in the ganglioside GM1 enzyme-linked immunosorbent assay (using either CT or Escherichia coli heat-labile toxin antitoxin) of both toxins indicates that 61892 toxin is antigenically indistinguishable from 569B CT. The specific activity of the two toxins in the rabbit ileal loop is virtually identical. Batch culture production of CT-like toxin and CT by isolates of V. mimicus and different biotypes of V. cholerae was found to be highest in shake flask cultures of Casamino Acids-yeast extract broth grown at 27 degrees C with vigorous aeration. Incorporation of lincomycin into the growth medium at a concentration of 50 micrograms/ml increased yields from wild-type strains. Dramatically higher yields were obtained when a spontaneous resistance mutant of strain 61892 was grown in the presence of 200 to 300 micrograms of lincomycin per ml. Under these conditions, yields of CT-like toxin were increased by 300- to 500-fold, and the highest yields reached more than 100 micrograms/ml after 44 h of culture. This is substantially higher than that reported in the literature for CT production by any strain of V. cholerae, including hypertoxigenic strain 569B.

Full text

PDF
501

Selected References

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

  1. Bhattacharya S., Bose A. K., Ghosh A. K. Permeability and enterotoxic factors of nonagglutinable vibrios Vibrio alcaligenes and Vibrio parahaemolyticus. Appl Microbiol. 1971 Dec;22(6):1159–1161. doi: 10.1128/am.22.6.1159-1161.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blake P. A., Allegra D. T., Snyder J. D., Barrett T. J., McFarland L., Caraway C. T., Feeley J. C., Craig J. P., Lee J. V., Puhr N. D. Cholera--a possible endemic focus in the United States. N Engl J Med. 1980 Feb 7;302(6):305–309. doi: 10.1056/NEJM198002073020601. [DOI] [PubMed] [Google Scholar]
  3. Craig J. P. A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera. Nature. 1965 Aug 7;207(997):614–616. doi: 10.1038/207614a0. [DOI] [PubMed] [Google Scholar]
  4. Craig J. P., Yamamoto K., Takeda Y., Miwatani T. Production of cholera-like enterotoxin by a Vibrio cholerae non-O1 strain isolated from the environment. Infect Immun. 1981 Oct;34(1):90–97. doi: 10.1128/iai.34.1.90-97.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DE S. N., CHATTERJE D. N. An experimental study of the mechanism of action of Vibriod cholerae on the intestinal mucous membrane. J Pathol Bacteriol. 1953 Oct;66(2):559–562. doi: 10.1002/path.1700660228. [DOI] [PubMed] [Google Scholar]
  6. Davis B. R., Fanning G. R., Madden J. M., Steigerwalt A. G., Bradford H. B., Jr, Smith H. L., Jr, Brenner D. J. Characterization of biochemically atypical Vibrio cholerae strains and designation of a new pathogenic species, Vibrio mimicus. J Clin Microbiol. 1981 Dec;14(6):631–639. doi: 10.1128/jcm.14.6.631-639.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Finkelstein R. A., Vasil M. L., Holmes R. K. Studies on toxinogenesis in Vibrio cholerae. I. Isolation of mutants with altered toxinogenicity. J Infect Dis. 1974 Feb;129(2):117–123. doi: 10.1093/infdis/129.2.117. [DOI] [PubMed] [Google Scholar]
  8. Guerrant R. L., Brunton L. L., Schnaitman T. C., Rebhun L. I., Gilman A. G. Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology: a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli. Infect Immun. 1974 Aug;10(2):320–327. doi: 10.1128/iai.10.2.320-327.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaper J. B., Bradford H. B., Roberts N. C., Falkow S. Molecular epidemiology of Vibrio cholerae in the U.S. Gulf Coast. J Clin Microbiol. 1982 Jul;16(1):129–134. doi: 10.1128/jcm.16.1.129-134.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaper J. B., Moseley S. L., Falkow S. Molecular characterization of environmental and nontoxigenic strains of Vibrio cholerae. Infect Immun. 1981 May;32(2):661–667. doi: 10.1128/iai.32.2.661-667.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelly M. T., Peterson J. W., Sarles H. E., Jr, Romanko M., Martin D., Hafkin B. Cholera on the Texas Gulf Coast. JAMA. 1982 Mar 19;247(11):1598–1599. [PubMed] [Google Scholar]
  12. Kusama H., Craig J. P. Production of Biologically Active Substances by Two Strains of Vibrio cholerae. Infect Immun. 1970 Jan;1(1):80–87. doi: 10.1128/iai.1.1.80-87.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Levner M., Wiener F. P., Rubin B. A. Induction of Escherichia coli and Vibrio cholerae enterotoxins by an inhibitor of protein synthesis. Infect Immun. 1977 Jan;15(1):132–137. doi: 10.1128/iai.15.1.132-137.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nishibuchi M., Seidler R. J., Rollins D. M., Joseph S. W. Vibrio factors cause rapid fluid accumulation in suckling mice. Infect Immun. 1983 Jun;40(3):1083–1091. doi: 10.1128/iai.40.3.1083-1091.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Oashi M., Shimada T., Fukumi H. In vitro production of enterotoxin and hemorrhagic principle by Vibrio cholerae, NAG. Jpn J Med Sci Biol. 1972 Jun;25(3):179–194. doi: 10.7883/yoken1952.25.179. [DOI] [PubMed] [Google Scholar]
  16. Rogers R. C., Cuffe R. G., Cossins Y. M., Murphy D. M., Bourke A. T. The Queensland cholera incident of 1977. 2. The epidemiological investigation. Bull World Health Organ. 1980;58(4):665–669. [PMC free article] [PubMed] [Google Scholar]
  17. Sack D. A., Huda S., Neogi P. K., Daniel R. R., Spira W. M. Microtiter ganglioside enzyme-linked immunosorbent assay for vibrio and Escherichia coli heat-labile enterotoxins and antitoxin. J Clin Microbiol. 1980 Jan;11(1):35–40. doi: 10.1128/jcm.11.1.35-40.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sack D. A., Sack R. B. Test for enterotoxigenic Escherichia coli using Y-1 adrenal cells in miniculture. Infect Immun. 1975 Feb;11(2):334–336. doi: 10.1128/iai.11.2.334-336.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Saunders D. W., Schanbacher K. J., Bramucci M. G. Mapping of a gene in Vibrio cholerae that determines the antigenic structure of cholera toxin. Infect Immun. 1982 Dec;38(3):1109–1116. doi: 10.1128/iai.38.3.1109-1116.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Singh S. J., Sanyal S. C. Enterotoxicity of the so-called NAG vibrios. Ann Soc Belg Med Trop. 1978;58(2):133–140. [PubMed] [Google Scholar]
  21. Tayot J. L., Holmgren J., Svennerholm L., Lindblad M., Tardy M. Receptor-specific large-scale purification of cholera toxin on silica beads derivatized with lysoGM1 ganglioside. Eur J Biochem. 1981 Jan;113(2):249–258. doi: 10.1111/j.1432-1033.1981.tb05060.x. [DOI] [PubMed] [Google Scholar]
  22. Yamamoto K., Takeda Y., Miwatani T., Craig J. P. Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin. Infect Immun. 1983 Mar;39(3):1128–1135. doi: 10.1128/iai.39.3.1128-1135.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zinnaka Y., Carpenter C. C., Jr An enterotoxin produced by noncholera vibrios. Johns Hopkins Med J. 1972 Dec;131(6):403–411. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES