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. 1982 Jan;43(1):39–49. doi: 10.1128/aem.43.1.39-49.1982

Partial Purification and Characterization of Hemolysin from a Psychrotrophic Kanagawa-Positive Marine Vibrio †

Susan Peters 1,‡, John A Baross 1, Richard Y Morita 1
PMCID: PMC241778  PMID: 16345927

Abstract

Psychrotrophic Kanagawa-positive marine vibrios were isolated from soft-shelled clams (Mya arenaria) collected in Yaquina Bay, Oreg. The 235 vibrio isolates obtained were screened for Gram reaction and morphology, Kanagawa reaction on Wagastsuma agar, and response to selected biochemical tests. The vibrio selected for further study was grown in broth, and the hemolysin was precipitated from a cleared supernatant with solid ammonium sulfate. The hemolytic substance was partially purified by DEAE-cellulose and Sephadex G-100 column chromatography. The hemolysin contained protein essential for activity, was thermolabile, and was more active against rabbit erythrocytes at 37°C than at lower temperatures. The molecular weight was estimated at 55,000 by using a Sephadex G-100 column. Hemolytic activity was partially inactivated by gangliosides and lowered against horse erythrocytes. The hemolysin did not react with antibody prepared against vibriolysin from Vibrio parahaemolyticus WP-1 by the Ouchterlony method. The hemolysin was high in aspartic and glutamic acids and low in arginine and histidine. Electrophoresis on a sodium dodecyl sulfate-polyacrylamide gel gave three major bands. The hemolysin from a psychrotrophic vibrio and the hemolytic exotoxin of V. parahaemolyticus had some similar and dissimilar characteristics. The possibility that a Vibrio sp. other than V. parahaemolyticus might serve as the reservoir for the Kanagawa phenotype is discussed.

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Selected References

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  1. Barker W. H., Jr, Hooper D., Baross J. A. Shellfish-related gastroenteritis. N Engl J Med. 1970 Aug 6;283(6):319–319. [PubMed] [Google Scholar]
  2. Baross J. A., Liston J., Morita R. Y. Ecological relationship between Vibrio parahaemolyticus and agar-digesting vibrios as evidenced by bacteriophage susceptibility patterns. Appl Environ Microbiol. 1978 Sep;36(3):500–505. doi: 10.1128/aem.36.3.500-505.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baross J. A., Liston J., Morita R. Y. Incidence of Vibrio parahaemolyticus bacteriophages and other Vibrio bacteriophages in marine samples. Appl Environ Microbiol. 1978 Sep;36(3):492–499. doi: 10.1128/aem.36.3.492-499.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baross J., Liston J. Occurrence of Vibrio parahaemolyticus and related hemolytic vibrios in marine environments of Washington State. Appl Microbiol. 1970 Aug;20(2):179–186. doi: 10.1128/am.20.2.179-186.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blake P. A., Merson M. H., Weaver R. E., Hollis D. G., Heublein P. C. Disease caused by a marine Vibrio. Clinical characteristics and epidemiology. N Engl J Med. 1979 Jan 4;300(1):1–5. doi: 10.1056/NEJM197901043000101. [DOI] [PubMed] [Google Scholar]
  6. Carpenter C. C. More pathogenic vibrios. N Engl J Med. 1979 Jan 4;300(1):39–41. doi: 10.1056/NEJM197901043000111. [DOI] [PubMed] [Google Scholar]
  7. Guerry P., Colwell R. R. Isolation of cryptic plasmid deoxyribonucleic acid from Kanagawa-positive strains of Vibrio parahaemolyticus. Infect Immun. 1977 Apr;16(1):328–334. doi: 10.1128/iai.16.1.328-334.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hansen W., Crokaert F., Yourassowsky E. Two strains of Vibrio species with unusual biochemical features isolated from ear tracts. J Clin Microbiol. 1979 Jan;9(1):152–153. doi: 10.1128/jcm.9.1.152-153.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Honda T., Taga S., Takeda T., Hasibuan M. A., Takeda Y., Miwatani T. Identification of lethal toxin with the thermostable direct hemolysin produced by Vibrio parahaemolyticus, and some physicochemical properties of the purified toxin. Infect Immun. 1976 Jan;13(1):133–139. doi: 10.1128/iai.13.1.133-139.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Mertens A., Nagler J., Hansen W., Gepts-Friedenreich E. Halophilic, lactose-positive Vibrio in a case of fatal septicemia. J Clin Microbiol. 1979 Feb;9(2):233–235. doi: 10.1128/jcm.9.2.233-235.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Miwatani T., Sakurai J., Yoshihara A., Takeda Y. Isolation and partial purification of thermolabile direct hemolysin of Vibrio parahaemolyticus. Biken J. 1972 Jun;15(2):61–66. [PubMed] [Google Scholar]
  13. Miwatani T., Takeda Y., Sakurai J., Yoshihara A., Taga S. Effect of heat (Arrhenius effect) on crude hemolysin of Vibrio parahaemolyticus. Infect Immun. 1972 Dec;6(6):1031–1033. doi: 10.1128/iai.6.6.1031-1033.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Morris D. L. Quantitative Determination of Carbohydrates With Dreywood's Anthrone Reagent. Science. 1948 Mar 5;107(2775):254–255. doi: 10.1126/science.107.2775.254. [DOI] [PubMed] [Google Scholar]
  15. Pezzlo M., Valter P. J., Burns M. J. Wound infection associated with Vibrio alginolyticus. Am J Clin Pathol. 1979 Apr;71(4):476–478. doi: 10.1093/ajcp/71.4.476. [DOI] [PubMed] [Google Scholar]
  16. Roland F. P. Leg gangrene and endotoxin shock due to vibrio parahaemolyticus--an infection acquired in New England coastal waters. N Engl J Med. 1970 Jun 4;282(23):1306–1306. doi: 10.1056/NEJM197006042822306. [DOI] [PubMed] [Google Scholar]
  17. Ryan W. J. Marine vibrios associated with superficial septic lesions. J Clin Pathol. 1976 Nov;29(11):1014–1015. doi: 10.1136/jcp.29.11.1014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sakurai J., Bahavar M. A., Jinguji Y., Miwatani T. Interaction of thermostable direct hemolysin of Vibrio parahaemolyticus with human erythrocytes. Biken J. 1975 Dec;18(4):187–192. [PubMed] [Google Scholar]
  19. Sakurai J., Matsuzaki A., Miwatani T. Purification and characterization of thermostable direct hemolysin of Vibrio parahaemolyticus. Infect Immun. 1973 Nov;8(5):775–780. doi: 10.1128/iai.8.5.775-780.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sakurai J., Matsuzaki A., Takeda Y., Miwatani T. Existence of two distinct hemolysins in Vibrio parahaemolyticus. Infect Immun. 1974 May;9(5):777–780. doi: 10.1128/iai.9.5.777-780.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Spark R. P., Fried M. L., Perry C., Watkins C. Vibrio alginolyticus wound infection: case report and review. Ann Clin Lab Sci. 1979 Mar-Apr;9(2):133–138. [PubMed] [Google Scholar]
  22. Spark R. P. Marine vibrio septicemia. N Engl J Med. 1979 Apr 26;300(17):989–989. doi: 10.1056/nejm197904263001718. [DOI] [PubMed] [Google Scholar]
  23. Takeda R., Honda T., Sakural J., Otomo N. Inhibition of hemolytic activity of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside. Infect Immun. 1975 Oct;12(4):931–933. doi: 10.1128/iai.12.4.931-933.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Takeda Y., Hori Y., Sakurai J., Miwatani T. Proceedings: Effect of heat on direct hemolysin of Vibrio parahaemolyticus; demonstration of an inactivating factor of purified thermostable direct hemolysin. Jpn J Med Sci Biol. 1975 Feb;28(1):90–93. [PubMed] [Google Scholar]
  25. Takeda Y., Takeda T., Honda T., Miwatani T. Inactivation of the biological activities of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside Gt1. Infect Immun. 1976 Jul;14(1):1–5. doi: 10.1128/iai.14.1.1-5.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zen-Yoji H., Hitokoto H., Morozumi S., Le Clair R. A. Purification and characterization o;f a hemolysin produced by Vibrio parahaemolyticus. J Infect Dis. 1971 Jun;123(6):665–667. doi: 10.1093/infdis/123.6.665. [DOI] [PubMed] [Google Scholar]

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