Study on Media Formulation and Production Process of Bacillus subtilis Spores for Animal Probiotics

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Wairuj Dechmahitkul
Chungeera Youkong
Kanokwan Poomputra
Saengchai Akeprathumchai
Phenjun Mekvijitsaeng

Abstract

The utilization of probiotic microorganism in animal feed has gained considerable attention becauseit helps improving animal health, increasing animal production and reducing antibiotic utilization. Bacillussubtillis (BS) is microorganism used in this study because it is a spore-forming bacteria and it can growrapidly. The aims of this study were to achieve high spore concentration by using media formulation andmode of cultivation. In addition, the antagonistic activity of BS against some pathogens was investigated.

Two sets of media formulations were studied in shake flask culture, i.e., molasses based formulaand defatted soy bean meal based formula. The molasses based media contained 19.85 g/L molasses (asreducing sugar), 0.35 g/L CaCI2 .2H2O and 0.15 g/L MnSO4 .H2O gave the highest spore concentrationof 5.5 x 109 CFU/ml whereas the soy bean meal based media contained 20 g/L soy bean meal, 3 g/Lmolasses and 0.5 g/L K2HPO4 gave the highest spore concentration of 1.3 x 109 CFU/ml. However, molassesbased media led to foaming problem in a fermentation process. Soy bean meal based media was thereforeused for fermentor cultivation. In 2 L-fermentor, spore concentrations at 48 h were 1.78 x 109 and4.03 x 108 CFU/ml for batch and fed-batch process respectively. Molasses added during cultivation couldenhance cell growth but it could impede spore formation. Therefore, in 75-L fed-batch cultivation, nomolasses was fed after the stationary phase of growth, resulting in the spore concentration of 5.73 x 109CFU/ml at 48 h. The results of well diffusion assay and paper disc method show that this probiotic doesnot have an antagonistic activity against several pathogens tested.

Keywords : Bacillus subtillis / Probiotics / Batch Cultivation / Fed-batch Cultivation /Media Formulation

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Author Biographies

Wairuj Dechmahitkul, King Mongkut's University of Technology Thonburi, Thakham, Bangkuntien, Bangkok 10150

Research Assistant, Biochemical Engineering Research and Pilot Plant Development Unit, BIOTECH.

Chungeera Youkong, King Mongkut's University of Technology Thonburi, Thakham, Bangkuntien, Bangkok 10150

Graduate Student, Bioresource and Technology Department.

Kanokwan Poomputra, King Mongkut's University of Technology Thonburi, Thakham, Bangkuntien, Bangkok 10150

Lecturer, Bioresource and Technology Department.

Saengchai Akeprathumchai, King Mongkut's University of Technology Thonburi, Thakham, Bangkuntien, Bangkok 10150

Lecturer, Bioresource and Technology Department.

Phenjun Mekvijitsaeng, King Mongkut's University of Technology Thonburi, Thakham, Bangkuntien, Bangkok 10150

Researcher, Pilot Plant Development and Training Institute.