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Soybean molasses-based bioindicator system for monitoring sterilization process: Designing and performance evaluation

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Abstract

A novel cost-effective Bacillus atrophaeus Sterilization Bioindicator System (BIS) with a high quality and performance was developed from a soybean byproduct and compared with the commercial BIS. It was composed of recovery medium and dry-fermented spores with sand as the support. The BIS was developed and optimized using a sequential experimental design strategy. The recovery medium contained soluble starch (1.0 g/L), soybean molasses (30.0 g/L), tryptone (40.0 g/L), and bromothymol blue (0.02 g/L) at pH 8.5. The solid-state fermentation conditions of the bioreactor and environmental humidity had no significant effects on the spore yield and dry-heat resistance. The only substrate mineral that showed a positive effect was Mn2+, allowing Mg2+, K+, and Ca2+ to be eliminated from the formulation. Validation of optimized medium indicated D 160°C = 6.8±1.0 min (3.6 min more than the minimum) and spore yield = 2.3 ± 0.5 × 109 CFU/g dry sand (10,000 × initial values). BIS performance resulted in D 160°C = 6.6 ± 0.1 min. Sporulation and germination kinetics allowed the sporulation process to be reduced to three days, and the growth of heat-damaged spores was sufficient to achieve visual identification of a non-sterile BIS within 21 h. Process economics was a minimum of 23.9%, and process cycle time was reduced from 29 to 15 days. The new BIS parameters demonstrated compliance to all regulatory requirements. No studies have yet described a BIS production from soybean molasses.

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Authors and Affiliations

  1. Production and Research Centre of Immunobiological Products, Parana State Department of Health, Piraquara-PR, 83302-200, Brazil

    Sandra Regina B. R. Sella & João Carlos Minozzo

  2. Luciana P. S. Vandenberghe, Carlos Ricardo Soccol Bioprocess Engineering and Biotechnology Department, Universidade Federal do Paraná, Curitiba-PR, 81531-990, Brazil

    Sandra Regina B. R. Sella, Carla Masetti, Luis Felipe M. Figueiredo, Luciana P. S. Vandenberghe & Carlos Ricardo Soccol

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  1. Sandra Regina B. R. Sella
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Correspondence to Sandra Regina B. R. Sella.

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Sella, S.R.B.R., Masetti, C., Figueiredo, L.F.M. et al. Soybean molasses-based bioindicator system for monitoring sterilization process: Designing and performance evaluation. Biotechnol Bioproc E 18, 75–87 (2013). https://doi.org/10.1007/s12257-012-0356-z

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  • DOI: https://doi.org/10.1007/s12257-012-0356-z

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