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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Application of Microbubbles Combining with Disinfectants to Inactivate Salmonella Typhimurium on Alfalfa Seeds and the Effects on Sprouting

by
Chih-Yao Hou
1,
Shih-Kao Chou
1,
Jong-Shinn Wu
2,
Hsiu-Ling Chen
3,
Pei-Wen Zhang
1,
Chih-Tung Liu
2,
Chun-Ping Hsiao
2 and
Chia-Min Lin
1,*
1
Department of Seafood Science, National Kaohsiung University of Science and Technology, Kaohsiung 81157, Taiwan
2
Department of Mechanical Engineering, National Yang Ming Chiao Tung University, Hsinchu 30010, Taiwan
3
Department of Food Safety/Hygiene and Risk Management, College of Medicine, National Cheng Kung University, Tainan 70403, Taiwan
*
Author to whom correspondence should be addressed.
Seeds 2025, 4(4), 51; https://doi.org/10.3390/seeds4040051 (registering DOI)
Submission received: 28 July 2025 / Revised: 3 October 2025 / Accepted: 23 October 2025 / Published: 24 October 2025
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Abstract

Microbial contamination is the main safety concern of sprouts and seeds are the major source. High concentrations of sanitizers (>10,000 mg/kg) are recommended for effective sanitation. Microbubble (MB) was reported to elevate sanitizer efficacy. Hence, MBs combined with disinfectants, chlorine dioxide (ClO2, 500 ppm), and slightly acidic electrolyzed water (SAEW, 250 ppm), were used to inactivate Salmonella Typhimurium on alfalfa seeds. After fulfilling MBs for 10 min, alfalfa seeds were washed in 10 L of water for 10, 20, or 30 min. Compared with untreated seeds, S. Typhimurium reductions obtained by SAEW-MBs (SMBs) and ClO2-MBs (CMBs) for 20 min were 3.8 and 3.3 log CFU/g, respectively. Conversely, the 20 min treatments of SAEW and ClO2 only obtained reductions of 0.9 and 1.1 log CFU/g, respectively. More surface ruptures on the seeds treated with CMBs were observed under a scanning electron microscope compared with the ones treated by water and ClO2 only. No adverse effects on the seed germination rate and the weight yield of sprouts were observed when treated with CMBs for 20 min. An MB device with capacity of 100 L was assembled and achieved reductions of 3.9 and 3.2 log CFU/g of natural microbes and S. Typhimurium, respectively, after 20 min CMB washing. Additionally, an MB device at 250 L was assembled and achieved 3.0 log CFU/g reduction in natural microbes. This study demonstrated that MBs enhanced the efficacy of disinfectants and could be applied in industrial-scale operations.
Keywords: alfalfa; microbubble; chlorine dioxide; electrolyzed water; Salmonella alfalfa; microbubble; chlorine dioxide; electrolyzed water; Salmonella
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MDPI and ACS Style

Hou, C.-Y.; Chou, S.-K.; Wu, J.-S.; Chen, H.-L.; Zhang, P.-W.; Liu, C.-T.; Hsiao, C.-P.; Lin, C.-M. Application of Microbubbles Combining with Disinfectants to Inactivate Salmonella Typhimurium on Alfalfa Seeds and the Effects on Sprouting. Seeds 2025, 4, 51. https://doi.org/10.3390/seeds4040051

AMA Style

Hou C-Y, Chou S-K, Wu J-S, Chen H-L, Zhang P-W, Liu C-T, Hsiao C-P, Lin C-M. Application of Microbubbles Combining with Disinfectants to Inactivate Salmonella Typhimurium on Alfalfa Seeds and the Effects on Sprouting. Seeds. 2025; 4(4):51. https://doi.org/10.3390/seeds4040051

Chicago/Turabian Style

Hou, Chih-Yao, Shih-Kao Chou, Jong-Shinn Wu, Hsiu-Ling Chen, Pei-Wen Zhang, Chih-Tung Liu, Chun-Ping Hsiao, and Chia-Min Lin. 2025. "Application of Microbubbles Combining with Disinfectants to Inactivate Salmonella Typhimurium on Alfalfa Seeds and the Effects on Sprouting" Seeds 4, no. 4: 51. https://doi.org/10.3390/seeds4040051

APA Style

Hou, C.-Y., Chou, S.-K., Wu, J.-S., Chen, H.-L., Zhang, P.-W., Liu, C.-T., Hsiao, C.-P., & Lin, C.-M. (2025). Application of Microbubbles Combining with Disinfectants to Inactivate Salmonella Typhimurium on Alfalfa Seeds and the Effects on Sprouting. Seeds, 4(4), 51. https://doi.org/10.3390/seeds4040051

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