Evaluation of factors influencing the growth of non-toxigenic Clostridium botulinum type E and Clostridium sp. in high-pressure processed and conditioned tender coconut water from Thailand
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Bacterial spores survive high pressure processing (HPP). Group II Clostridium botulinum is an obligate anaerobe spore-forming pathogen that can produce the botulinum neurotoxin under refrigeration. This study assessed nontoxigenic type E C. botulinum and Group II Clostridium sp. growth in raw and HPP (550 MPa, 3 min, 10 °C) Thai coconut water (CCW; pH 5.2). No spore germination or growth occurred in HPP CCW inoculated with 105 CFU/ml after 61 days regardless of oxygen concentration (<0.5 – 11 mg/l) or storage temperature (4 and 20 °C). Spore concentration decreased by 3.0 ± 0.1 log CFU/ml in a worst-case scenario consisting of non-HPP filter-sterilized CCW (pH 7.0) under anoxic incubation at 30 °C during 61 days, suggesting spore germination followed by cellular death. Supplementing filter-sterilized CCW (pH 7.0) with selected germinants and free amino acids did not support spore development, but the addition of nutrient-rich laboratory media (TPGY broth) at low concentrations (6.25%) promoted growth, suggesting that a lack of nutrients prevents C. botulinum development in CCW. Further risk assessment will require evaluating other CCW varieties and toxin production.
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HHP is not considered a validated process for eliminating the spores of Clostridium botulinum in low-acid products (pH > 4.6), including CW with a pH range between 4.8 and 5.7.
“Unfortunately, HHP is not considered a validated process for eliminating the spores of Clostridium botulinum in low-acid products (pH > 4.6), including CW with a pH range between 4.8 and 5.7 [59].”
From Research Progress in Coconut Water: A Review of Nutritional Composition, Biological Activities, and Novel Processing Technologies (2025) · cites this paper as [59]
CW is deficient in some essential nutrients required by C. botulinum to grow.
“By supplementing CW with selected germinants, amino acids, and nutrient-rich laboratory media (TPGY broth), the authors concluded that CW was deficient in some essential nutrients required by C. botulinum to grow.”
From Research Progress in Coconut Water: A Review of Nutritional Composition, Biological Activities, and Novel Processing Technologies (2025) · cites this paper as [59]
After HHP treatment at 550 MPa and 10 °C for 3 min, the total counts of inoculated non-toxigenic C. botulinum spores in CW were not significantly reduced and remained constant during 61 days of storage at 4 °C, regardless of initial dissolved oxygen content.
“After HHP treatment at 550 MPa and 10 °C for 3 min, the total counts of inoculated non-toxigenic C. botulinum spores in CW were not significantly reduced and remained constant during the 61 days of storage at 4 °C, regardless of the initial dissolved oxygen content [59].”
From Research Progress in Coconut Water: A Review of Nutritional Composition, Biological Activities, and Novel Processing Technologies (2025) · cites this paper as [59]
Storage of HHP-treated CW at 10 and 20 °C showed no growth of C. botulinum, but there was a significant increase in total aerobic counts within 10 and 4 days, respectively, suggesting limited capacity of HHP to inactivate all spoilage microorganisms.
“Storage of HHP-treated CW at 10 and 20 °C also showed no growth of C. botulinum, but there was a significant increase in total aerobic counts within 10 and 4 days, respectively, suggesting the limited capacity of HHP to inactivate all spoilage microorganisms.”
From Research Progress in Coconut Water: A Review of Nutritional Composition, Biological Activities, and Novel Processing Technologies (2025) · cites this paper as [59]
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References 71 · Cited by 23
- Authors
- González-Angulo, M., Clauwers, C., Harastani, R., Tonello, C., Jaime, I., Rovira, J., & Michiels, C. W.
- Affiliations
- Universidad de Burgos (ES)
- KU Leuven (BE)
- Corresponding author
- González-Angulo, Mario
- Venue
- Food Research International
- Publisher
- Elsevier BV
- Published
- 2020-08
- Volume
- 134
- Pages
- 109278
- ISSN
- 0963-9969 (ISSN-L)
- OpenAlex
- W3022650741
- Cited by
- 23
- References
- 71
- Citations by year
- 2026: 3 · 2025: 1 · 2024: 3 · 2023: 5 · 2022: 5 · 2021: 3 · 2020: 3
- FWCI
- 2.07
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Food Research International (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Repositorio Institucional de la Universidad de Burgos (University of Burgos) (submittedVersion, cc-by)
- Lirias (submittedVersion, other-oa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Microbial Inactivation Methods
- Health Sciences › Medicine › Neurology › Botulinum Toxin and Related Neurological Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Listeria monocytogenes in Food Safety
- Keywords
- Clostridium botulinum type E, coconut water, high pressure processing, spore germination, spore survival, bacterial growth, oxygen concentration, storage temperature, germinants, free amino acids, nutrient limitation, food safety, risk assessment
- MeSH terms
- Clostridium, Cocos, Thailand, Water, Clostridium botulinum type E
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