Cereal Research Communications

, Volume 44, Issue 1, pp 131–140 | Cite as

Spray-dried Powder of Bacillus megaterium for Control of Rice Sheath Blight Disease: Formulation Protocol and Efficacy Testing in Laboratory and Greenhouse

  • A. ChumthongEmail author
  • R. Wiwattanapatapee
  • H. Viernstein
  • A. Pengnoo
  • M. Kanjanamaneesathian


A spray-dried powder containing Bacillus megaterium was developed and tested for control of rice sheath blight disease in the greenhouse. The formulation consisted of 20 ml of an endospore suspension of B. megaterium, 20% w/v of skim milk powder and 1.25% w/v of polyvinyl pyrrolidone k-90, that were mixed and spray dried at 120 °C. The powder displayed good physical characteristics, such as a low-moisture content and a high solubility in water. Bacterial viability in the powder was 3.5±0.1 × 1011 cfu/g after production and remained relatively stable (at 2.2±0.1 × 1010 cfu/g) after 12 months of storage at room temperature. In the laboratory, a 0.1% (w/v) aqueous solution of the formulation was effective in inhibiting the mycelia growth of Rhizoctonia solani (98.5±0.1% inhibition). Under greenhouse conditions, a 0.1% (w/v) aqueous solution applied by either spraying 1 day before inoculating R. solani or spraying 1, 7 and 15 days after inoculation of rice plants with R. solani was more effective in suppressing sheath blight disease than the blank formulation but was less effective than a chemical fungicide control.


Bacillus megaterium bio-pesticide formulation rice disease spray-dried powder 


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We would like to thank Dr. Brian Hodgson for assistance with the English.


  1. Abadias, M., Benabarre, A., Teixido, N., Usall, J., Vinas, I. 2001. Effect of freeze-drying and protectants on viability of the biocontrol yeast Candida sake. Int. J. of Food Microbiol. 65:173–182.CrossRefGoogle Scholar
  2. Ananta, E., Volkert, M., Knorr, D. 2005. Cellular injuries and storage stability of spray-dried Lactobacillus rhamnosus GG. Int. Dairy J. 15:399–409.CrossRefGoogle Scholar
  3. Anh, S.W., de La Pena, R., Candole, B.L., Mew, T.W. 1986. A new scale for rice sheath blight disease assessment. International Rice Research Institute. Manila, Philippines. pp. 11, 17.Google Scholar
  4. Boza, Y., Barbin, D., Scamparini, A.R.P. 2004. Effect of spray-drying on the quality of encapsulated cells of Beijerinckia sp. Process Biochem. 39:1275–1284.CrossRefGoogle Scholar
  5. Burges, H.D. 1998. Formulation of mycoinsecticides. In: Burges, H.D. (ed.), Formulation of Microbial Biopesticides: Beneficial microorganism, Nematodes and Seed treatments, Kluwer Academic Publishers. Dordrecht, The Netherlands. pp. 131–187.CrossRefGoogle Scholar
  6. Bühler, V. 2001. Kollidon: Polyvinyl Pyrrolidone for the Pharmaceutical Industry. 6th Ed. BASF, Pharma Ingredients. Ludwigshafen, Germany. pp. 139–157.Google Scholar
  7. Chumthong, A., Kanjanamaneesathian, M., Pengnoo, A., Wiwattanapatapee, R. 2008. Water-soluble granules containing Bacillus megaterium for biological control of rice sheath blight: Formulation, bacterial viability and efficacy testing. World J. of Microbiol. and Biotechnol. 24:2499–2507.CrossRefGoogle Scholar
  8. Conrad, P.B., Miller, D.P., Cielenski, P.R., de Pablo, J.J. 2000. Stabilization and preservation of Lactobacillus acidophilus in saccharide matrices. Crybiology 41:17–24.CrossRefGoogle Scholar
  9. Desmond, C., Stanton, C., Fitzgerald, G.F., Collins, K., Ross, R.P. 2001. Environmental adaptation of probiotic lactobacilli towards improvement of performance during spray drying. Int. Dairy J. 11:801–808.CrossRefGoogle Scholar
  10. Gamliel, A., Katan, J., Cohen, E. 1989. Toxicity of chloronitrobenzenes to Fusarium oxysporum and Rhizoctonia solani as related to their structure. Phytoparasitica 17:101–106.CrossRefGoogle Scholar
  11. Gnanamanickam, S.S., Candole, B.L., Mew, T.W. 1992. Influence of soil factors and cultural practice on biological control of sheath blight of rice with antagonistic bacteria. Plant and Soil 144:67–75.CrossRefGoogle Scholar
  12. Horaczek, A., Viernstein, H. 2004. Comparison of three commonly used drying technologies with respect to activity and longevity of aerial conidia of Beauveria brongniartii and Metarhizium anisopliae. Biol. Control 31:65–71.CrossRefGoogle Scholar
  13. Johnson, J.A.C., Etzel, M.R. 1993. Inactivation of lactic acid bacteria during spray-drying. Aiche Symposium 89:89–107.Google Scholar
  14. Johnson, J.A.C., Etzel, M.R. 1995. Properties of Lactobacillus helveticus CNRZ-32 attenuated by spray-drying, freeze-drying, or freezing. J. Dairy Sci. 78:761–768.CrossRefGoogle Scholar
  15. Kanjanamaneesathian, M., Chumthong, A., Pengnoo, A., Wiwattanapatapee, R. 2009. Bacillus megaterium suppresses major Thailand rice diseases. Asian J. of Food and Agro-Industry S:154–159.Google Scholar
  16. Kanjanamaneesathian, M., Kusonwiriyawong, C., Pengnoo, A., Nilratana, L. 1998. Screening of potential bacterial antagonists for control of sheath blight in rice and development of suitable bacterial formulations for effective application. Australasian Plant Pathol. 27:198–206.CrossRefGoogle Scholar
  17. Kumar, K.V.K., Yellareddygari, S.K.R., Reddy, M.S., Kloepper, .J.W., Lawrence, K.S., Miller, M., Sudini, H., Surendranatha, Reddy, E.C., Zhou, X.G., Groth, D.E. 2013. Ultrastructural studies on the interaction between Bacillus subtilis MBI 600 (Integral) and the rice sheath blight pathogen, Rhizoctonia solani. African J. of Microbiol. Res. 7:2078–2086.CrossRefGoogle Scholar
  18. Lian, W.C., Hsiao, H.C., Chou, C.C. 2002. Survival of bifidobacteria after spray-drying. J. Food Microbiol. 74:79–86.CrossRefGoogle Scholar
  19. Mosquera-Espinosa, A.T., Bayman, P., Prado, G.A., Gómez-Carabalí, A., Otero, J.T. 2013. The double life of Ceratobasidium: orchid mycorrhizal fungi and their potential for biocontrol of Rhizoctonia solani sheath blight of rice. Mycologia 105:141–150.CrossRefGoogle Scholar
  20. Naeimi, S., Okhovvat, S.M., Javan-Nikkhah, M., Vágvölgyi, C., Khosravi, V., Kredics, L. 2010. Biological control of Rhizoctonia solani AG1–1A, the causal agent of rice sheath blight with Trichoderma strains. Phytopathologia Mediterranea 49:287–300.Google Scholar
  21. Savary, S., Willocquet, L., Elazegui, F.A., Castilla, N.P., Teng, P.S. 2000. Rice pest constraints in tropical Asia: quantification of yield losses due to rice pests in a range of production situations. Plant Dis. 84:357–369CrossRefGoogle Scholar
  22. Selmer-Olsen, E., Sorhaug, T., Birkeland, S.E., Pehrson, R. 1999. Survival of Lactobacillus helveticus entrapped in Ca-alginate in relation to water content, storage and rehydration. J. of Industrial Microbiol. Biotechnol. 23:79–85.CrossRefGoogle Scholar
  23. To, B.C.S., Etzel, M.R. 1997a. Spray drying, freeze drying, or freezing of three different lactic acid bacteria species. J. Food Sci. 62:576–578.CrossRefGoogle Scholar
  24. To, B.C.S., Etzel, M.R. 1997b. Survival of Brevibacterium linens ATCC 9174 after spray drying, freeze drying, or freezing. J. Food Sci. 62:167–170.CrossRefGoogle Scholar
  25. Wiwattanapatapee, R., Chumthong, A., Pengnoo, A., Kanjanamaneesathian, M. 2007. Effervescent fast-disintegrating bacterial formulation for biological control of rice sheath blight. J. of Controlled Release 119:229–235.CrossRefGoogle Scholar
  26. Yánez-Mendizábal, V., Viñas, I., Usall, J., Torres, R., Solsona, C., Abadias, M., Teixidó, N. 2012. Formulation development of the biocontrol agent Bacillus subtilis strain CPA-8 by spray-drying. J. Appl. Microbiol. 112:954–965.CrossRefGoogle Scholar
  27. Zamora, L.M., Carretero, C., Pares, D. 2006. Comparative survival rates of lactic acid bacteria isolated from blood, following spray-drying and freeze-drying. Food Sci. and Technol. Int. 12:77–84.CrossRefGoogle Scholar
  28. Zuberer, D.A. 1994. Recovery and enumeration of viable bacteria. In: Weaver, R.W., Angle, S., Bottomley, P., Bezdicek, D., Smith, S., Tabatabai, A., Wollum, A. (eds), Methods of Soil Analysis: Part 2, Microbiological and Biochemical Properties, Number 5. In Soil Science Society of America Book Series. Soil Science Soc. of America Inc. Wisconsin, USA. pp. 119–143.Google Scholar

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© Akadémiai Kiadó, Budapest 2016

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Authors and Affiliations

  • A. Chumthong
    • 1
    Email author
  • R. Wiwattanapatapee
    • 2
  • H. Viernstein
    • 3
  • A. Pengnoo
    • 4
  • M. Kanjanamaneesathian
    • 5
  1. 1.Department of Agricultural Technology, Faculty of Agricultural TechnologySongkhla Rajabhat UniversityMuang, SongkhlaThailand
  2. 2.Department of Pharmaceutical Technology, Faculty of Pharmaceutical SciencesPrince of Songkla UniversityHat Yai, SongkhlaThailand
  3. 3.Department of Pharmaceutical Technology and Biopharmaceutics, Faculty of Life SciencesUniversity of ViennaViennaAustria
  4. 4.Department of Earth Sciences, Faculty of Natural ResourcesPrince of Songkla UniversityHatYai, SongkhlaThailand
  5. 5.Plant Production Technology Program, Faculty of Animal Science and Agricultural TechnologySilpakorn University, Phetchaburi IT campusCha-Am, PhetchaburiThailand

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