Journal of Industrial Microbiology

, Volume 13, Issue 2, pp 112–119 | Cite as

Fermentation and toxin studies of the molluscicidal strains ofBacillus brevis

  • Samuel Singer
  • Thomas B. Bair
  • Terry B. Hammill
  • Aminata Maman Berte
  • Margarita M. Correa-Ochoa
  • Angela D. Stambaugh


Strain SS86-4 was one of 40Bacillus brevis strains shown to be molluscicidal to the schistosomiasis snail vectorBiomphalaria glabrata. When grown in mB4 medium in 2-L fermentors, SS86-4 was molluscicidal only if fructose or phenylalanine was present in the medium. This is reminiscent of secondary fermentation factor effects, in this case an antioxidant effect. In vivo proteases also were capable of reducing molluscicidal activity. The molluscicidal toxin has an LC50 of 1 μg toxin protein ml−1 (approx. 1 p.p.m.) and may be described as a small proteinaceous, heat-stable, oxygen-sensitive entity associated with the particulate portion of the cell wall fraction ofB. brevis that is formed prior to sporulation. Initial information indicates that its HPLC signature shows major peaks at 148.37 and 163.96 s and consists of two bands of approximately 5.3 kDa and 8.7 kDa on PAGE gel.

Key words

MolluscicidalBacillus toxin Bacillus brevis Biomphalaria glabrata Biocontrol of snails Antioxidant preservation of toxin Secondary fermentation factor 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Agathos, S.N. and A.L. Demain. 1983. Optimization of fermentation processes through the control of in vivo inactivation of microbial biosynthetic enzymes. In: Foundations of Biochemical Engineering (Blanch, H.W., E.T. Papoutsakis and G. Stephanopoulos, eds), pp. 53–67, American Chemical Society, Washington, DC.Google Scholar
  2. 2.
    Agathos, S.N. and A.L. Demain. 1986. Substrate amino acid-mediated stabilization of gramicidin S synthetase activity against inactivation in vivo. Enzyme Microb. Technol. 8: 465–468.Google Scholar
  3. 3.
    Baumann, P., M.A. Clark, L. Baumann and A.H. Broadwell. 1991.Bacillus sphaericus as a mosquito pathogen: properties of the organism and its tovins. Bacteiol. Rev. 55: 425–436.Google Scholar
  4. 4.
    Bone, L.W., K.P. Bottjer and S.S. Gill. 1986.Trichostrongylus colubriformis: isolation and characterization of ovicidal activity fromBacillus thuringiensis isrealensis. Exp. Parasitology 62: 247–253.Google Scholar
  5. 5.
    Bradford, M.M. 1976. A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.Google Scholar
  6. 6.
    Cheng, T.C. 1974. Molluscicides in Schistosomiasis Control. Academic Press, New York.Google Scholar
  7. 7.
    Davidson, E.W. 1983. Alkaline extraction of toxin from spores of the mosquito pathogen,Bacillus sphaericus strain 1593. Can. J. Microbiol. 29: 271–275.Google Scholar
  8. 8.
    Kreig, A. and H.G. Miltenburger. 1984. Bioinsecticides: 1.Bacillus thuringiensis. Adv. Biotechnol. Proc. 3: 273–290.Google Scholar
  9. 9.
    Luthy, P., H.R. Ebersold, J.L. Cordier and H.M. Fischer. 1985. Insecticidal metabolites of spore-forming bacilli. In: Fundamental and Applied Aspects of Bacterial Spores (Dring, G.J., D.J. Ellar and G.W. Gould, eds), pp. 475–484, Academic Press, London.Google Scholar
  10. 10.
    Margalith, P. 1964. Secondary factors in fermentation processes. Adv. Appl. Microbiol. 6: 69–90.Google Scholar
  11. 11.
    McCullogh, F.S. and K.E. Mott. 1983. The role of molluscicides in schistosomiasis control. Document WHO/SCHISTO/83.72.Google Scholar
  12. 12.
    Myers, P.S. and A.A. Youston. 1980. Localization of a mosquito-larval toxin ofBacillus sphaericus 1593. Appl. Environ. Microbiol. 39: 1205–1211.Google Scholar
  13. 13.
    Schagger, H. and G. von Jagow. 1987. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166: 368–379.Google Scholar
  14. 14.
    Singer, S. 1979. Use of entomogenous bacteria against insects of public health importance. Devel. Indust. Microbiol. 20: 117–122. Society for Industrial Microbiology, Arlington, Virginia.Google Scholar
  15. 15.
    Singer, S. 1987. Current status of the microbial larvicideBacillus sphaericus. In: Biotechnology Advances in Insect Pathology and Cell Culture (Maramorosch, K., ed.), pp. 133–163, Academic Press, New York.Google Scholar
  16. 16.
    Singer, S., K.A. Doherty and A.D. Stambaugh. 1988. Preliminary examination of molluscicidal strains of the genus Bacillus. 88th Annual Meeting American Society for Microbiology, Miami, Florida (Abstr., p. 76).Google Scholar
  17. 17.
    Stecher, P.G., M.J. Finkel, O.H. Siegmund and B.M. Zafranski. 1960. The Merck Index of Chemicals and Drugs, p. 497, Merck and Co. Inc., Rahway.Google Scholar
  18. 18.
    Williston, B.K. and S. Singer. 1987. Initial studies of the use of aminopeptidases for the differentiation ofBacillus sphaericus strains. J. Ind. Microbiol. 2: 285–292.Google Scholar
  19. 19.
    World Health Organization. 1984. Report of an informal consultation on research on the biological control of snail intermediate hosts. Document TDR/BCV-SCH/SIH/84.3, Geneva.Google Scholar

Copyright information

© Society for Industrial Microbiology 1994

Authors and Affiliations

  • Samuel Singer
    • 1
  • Thomas B. Bair
    • 1
  • Terry B. Hammill
    • 1
  • Aminata Maman Berte
    • 1
  • Margarita M. Correa-Ochoa
    • 1
  • Angela D. Stambaugh
    • 1
  1. 1.Department of Biological SciencesWestern Illinois UniversityMacombUSA

Personalised recommendations