Development of W/O/W Multiple Emulsion Formulation Containing Burkholderia gladioli

  • Published : 2005.02.01

Abstract

W/O/W (water-in-oil-in-water) type multiple emulsion was applied to improve the storage stability of an antagonistic microorganism, Burkholderia gladioli. Encapsulation of microorganism into a W/O/W emulsion was conducted by using a two-step emulsification method. W/O/W emulsion was prepared by the incorporation of B. gladioli into rapeseed oil and the addition of polyglycerin polyriconolate (PGPR) and castor oil polyoxyethylene (COG 25) as the primary and secondary emulsifier, respectively. Microcrystalline cellulose was used as an emulsion stabilizer. To evaluate the usefulness of W/O/W emulsion formulation as a microbial pesticide for controlling the bacterial wilt pathogen (Ralstonia solanacearum), the storage stability and antagonistic activity of emulsion formulation were tested in vitro. The storage stability test revealed that the viability of formulated cells in emulsion was higher than that of unformulated cells in culture broth. At $4^{\circ}C$, the viabilities of formulated cells and unformulated cells at the end of 20 weeks decreased to about 2 and 5 log cycles, respectively. At $37^{\circ}C$, the viability of formulated cells decreased to only 2 log cycles at the end of storage. On the other hand, the viable cells in culture broth were not detected after 13 weeks. In activity test, formulated cells in emulsion were more effective in inhibiting the growth of pathogen than unformulated cells in culture broth. Unformulated cells completely lost their antagonistic activity during storage under similar conditions. The W/O/W multiple emulsion formulation was shown to be useful as the novel liquid formulation for biological control.

Keywords

References

  1. Alves, R. T., R. P. Bateman, C. Prior, and S. R. Leather. 1998. Effects of simulated solar radiation on conidial germination of Metarhizium anisopliae in different formulations. Crop Prot. 17: 675- 679 https://doi.org/10.1016/S0261-2194(98)00074-X
  2. Amsellern, Z., N. K. Zidack, P. C. Quimby Jr., and J. Gressel. 1999. Long-term dry preservation of viable mycelia of two mycoherbicidal organisms. Crop Prot. 18: 643- 649 https://doi.org/10.1016/S0261-2194(99)00070-8
  3. Bagnasco, P., L. de la Fuente, G. Gualtieri, F. Noya, and A. Arias. 1998. Fluorescent Pseudomonas spp. as biocontrol agents against forage legume root pathogenic fungi. Soil Biol. Biochem. 30: 1317- 1322 https://doi.org/10.1016/S0038-0717(98)00003-0
  4. Bae, E. K., Y. Cho, and J. Park. 2004. Studies on the development of microbial cryoprotectant formulation using a W/O/W multiple emulsion system. J. Microbiol. Biotechnol. 14: 673- 679
  5. Burges, H. D. 1998. Formulation of Microbial Biopesticides. London: Kluwer Academic Publishers, pp. 21- 26
  6. Cotxarrera, L., M. I. Trillas-Gay, C. Steinberg, and C. Alabouvette. 2002. Use of sewage sludge compost and Trichoderma asperellum isolates to suppress Fusarium wilt of tomato. Soil Biol. Biochem. 34: 467-476 https://doi.org/10.1016/S0038-0717(01)00205-X
  7. Garti, N. 1997. Progress in stabilization and transport phenomena of double emulsions in food applications. Lebensm.-Wiss. u-Technol. 30: 222- 235 https://doi.org/10.1006/fstl.1996.0176
  8. Hino, T., Y. Kawashima, and S. Shimabayashi. 2000. Basic study for stabilization of w/o/w emulsion and its application to transcatheter arterial embolization therapy. Advanced Drug Delivery Reviews 45: 27-45 https://doi.org/10.1016/S0169-409X(00)00098-3
  9. Kim, J. W., J. G. Kim, B. K Park, O. H. Choi, C. S. Park, and I. G. Hwang. 2003. Identification of genes for biosynthesis of antibacterial compound from Pseudomonas fluorescens B16 and its activity against Ralstonia solanacearum. J. Microbiol. Biotechnol. 13: 292- 300
  10. Kim, Y. S., H. S. Lim, and S. D. Kim. 1994. Bacillus subtilis YB-70 as a biocontrol agent of Fusarium solani causing plant root-rot. J. Microbiol. Biotechnol. 4: 68- 74
  11. Klein, T. A., B. A. Auld, and F. Wang. 1995. Evaluation of oil suspension emulsions of Colletotrichum orbiculare as a mycoherbicide in field trials. Crop Prot. 14: 193-197 https://doi.org/10.1016/0261-2194(95)00009-B
  12. Koch, E. 1999. Evaluation of commercial products for microbial control of soil-borne plant diseases. Crop Prot. 18: 119-125 https://doi.org/10.1016/S0261-2194(98)00102-1
  13. Li, W., D. P. Roberts, P. D. Dery, S. L. F. Meyer, S. Lohrke, R. D. Lumsden, and K. P. Hebbar, 2002. Broad spectrum antibiotic activity and disease suppression by the potential biocontrol agent Burkholderia ambifaria BC-F. Crop Prot. 21: 129-135 https://doi.org/10.1016/S0261-2194(01)00074-6
  14. Lim, H. S., J. M. Lee, and S. D. Kim. 2003. A Plant growth-promoting Pseudomonas fluorescens GL20 - mechanism for disease suppression, outer membrane receptors for ferric siderophore, and genetic improvement for increased biocontrol efficacy. J. Microbiol. Biotechnol. 12: 249-257
  15. Nonomura, T., Y. Matsuda, M. Bingo, M. Onishi, K. Matsuda, S. Harada, and H. Toyoda. 2001. Algicidal effect of 3-(3-indolyl) butanoic acid, a control agent of the bacterial wilt pathogen, Ralstonia solanacearum. Crop Prot. 20: 935-939 https://doi.org/10.1016/S0261-2194(01)00048-5
  16. Okochi, H. and M. Nakano. 2000. Preparation and evaluation of w/o/w type emulsions containing vancomycin. Advanced Drug Delivery Reviews 45: 5- 26 https://doi.org/10.1016/S0169-409X(00)00097-1
  17. Quimby, Jr. P. C, J. L. Birdsall, A. J. Caesar, W. J. Connick Jr., C. D. Boyette, T. C. Caesar, and D. C. Sands. 1994. Oil and absorbent coated granules containing encapsulated living organisms for controlling agricultural pests. U.S. patent no. 5,358,863
  18. Risch, S. J. and G. A. Reineccius. 1988. Spray dried orange oil-effect of emulsion size on flavor retention and shelf stability, pp. 67- 77. In Risch, S. J. and Reineccius, G. A. (eds.), Flavor Encapsulation. ACS symposium series 370, American Chemical Society, Illinois
  19. Sabaratnam, S. and J. A. Traquair. 2002. Formulation of a Streptomyces biocontrol agent for the suppression of Rhizoctonia damping-off in tomato transplants. Biol. Control 23: 245-253 https://doi.org/10.1006/bcon.2001.1014
  20. Shasha, B. S. and R. L. Dunkle. 1989. Starch encapsulation of entomopathogens. U.S. patent no. 4,859,377
  21. Shigemitsu, H. 1987. Immobilization of microorganisms antagonistic to plant pathogenic microorganisms. U.S. patent no. 4,647,537
  22. Shimizu, M. and Y. Nakane. 1995. Encapsulation of biologically active proteins in a multiple emulsion. Biosci. Biotech. Biochem. 59: 492-496 https://doi.org/10.1271/bbb.59.492
  23. Silva-Cunha, A., J. L. Grossiord, F. Pusieux, and M. Seiller. 1997. W/O/W multiple emulsions of insulin containing a protease inhibitor and and absorption enhancer: Preparation, characterization and determination of stability towards proteases in vitro. Int. J. Pharmaceutics 158: 79-89 https://doi.org/10.1016/S0378-5173(97)00249-4
  24. Silver, S. C. 1999. Granular formulation of biological entities with improved storage stability. U.S. patent no. 5,965,149
  25. Stirling, G. R., K. A. Licastro, L. M. West, and L. J. Smith. 1998. Development of commercially acceptable formulations of the nematophagous fungus Verticillium chlamydosporium. Biol. Control 11: 217-223 https://doi.org/10.1006/bcon.1997.0602
  26. Vidhyasekaran, P, K. Sethuraman, K. Rajappan, and K. Vasumathi. 1997. Powder formulations of Pseudomonas fluorescens to control pigeonpea wilt. Biol. Control 8: 166-171 https://doi.org/10.1006/bcon.1997.0511
  27. Yoon, C. S., K. W. Yu, S. H. Bae, H. H. Song, H. S. Park, and C. Lee. 2003. Chemical properties and physiological activities of synnemata of Beauveria hassiana. J. Microbiol. Biotechnol. 13: 125- 133