Optimization of Spore Purification and Quality Assessment in Paenibacillus Species

To enhance the consistency and quality of Paenibacillus spore preparations used in food safety research, we examined two widely adopted purification methods—lysozyme treatment and HistDenz gradient centrifugation—under different sporulation conditions (solid agar and liquid media). Spores from four...

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Main Authors: Souichirou Kawai, Miyo Nakano
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Journal of Food Protection
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Online Access:http://www.sciencedirect.com/science/article/pii/S0362028X25001474
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author Souichirou Kawai
Miyo Nakano
author_facet Souichirou Kawai
Miyo Nakano
author_sort Souichirou Kawai
collection DOAJ
description To enhance the consistency and quality of Paenibacillus spore preparations used in food safety research, we examined two widely adopted purification methods—lysozyme treatment and HistDenz gradient centrifugation—under different sporulation conditions (solid agar and liquid media). Spores from four Paenibacillus strains (P. odorifer JCM 21743T and JCM 13339, P. terrae JCM 11466T, and P. polymyxa JCM 2507T) were purified and evaluated for metabolic activity using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) staining and for germination capacity through growth curve analysis. CTC staining verified the complete absence of metabolically active vegetative cells in all purified samples. Growth assays demonstrated that lysozyme treatment reduced spore viability in certain strains, whereas HistDenz purification more reliably preserved germination potential. Additionally, spores generated on solid medium displayed improved growth performance—evidenced by higher peak turbidity and shorter doubling times—compared to those produced in liquid medium for selected strains. These results demonstrate that both the purification technique and sporulation environment have a significant impact on the physiological quality of Paenibacillus spores. Therefore, selecting strain-specific preparation protocols is essential for accurately assessing spore viability and ensuring their effective application in downstream detection or safety assessments.
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spelling doaj-art-ccbe8fabace7403cb6cb3f40656f11992025-08-20T05:04:16ZengElsevierJournal of Food Protection0362-028X2025-09-01881010059510.1016/j.jfp.2025.100595Optimization of Spore Purification and Quality Assessment in Paenibacillus SpeciesSouichirou Kawai0Miyo Nakano1Corresponding author.; Toyo Institute of Food Technology, 23-2, 4-chome, Minami-hanayashiki, Kawanishi, Hyogo 666-0026, JapanToyo Institute of Food Technology, 23-2, 4-chome, Minami-hanayashiki, Kawanishi, Hyogo 666-0026, JapanTo enhance the consistency and quality of Paenibacillus spore preparations used in food safety research, we examined two widely adopted purification methods—lysozyme treatment and HistDenz gradient centrifugation—under different sporulation conditions (solid agar and liquid media). Spores from four Paenibacillus strains (P. odorifer JCM 21743T and JCM 13339, P. terrae JCM 11466T, and P. polymyxa JCM 2507T) were purified and evaluated for metabolic activity using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) staining and for germination capacity through growth curve analysis. CTC staining verified the complete absence of metabolically active vegetative cells in all purified samples. Growth assays demonstrated that lysozyme treatment reduced spore viability in certain strains, whereas HistDenz purification more reliably preserved germination potential. Additionally, spores generated on solid medium displayed improved growth performance—evidenced by higher peak turbidity and shorter doubling times—compared to those produced in liquid medium for selected strains. These results demonstrate that both the purification technique and sporulation environment have a significant impact on the physiological quality of Paenibacillus spores. Therefore, selecting strain-specific preparation protocols is essential for accurately assessing spore viability and ensuring their effective application in downstream detection or safety assessments.http://www.sciencedirect.com/science/article/pii/S0362028X25001474CTC stainingFlow cytometryHistDenz treatmentLysozyme treatmentSpore purification
spellingShingle Souichirou Kawai
Miyo Nakano
Optimization of Spore Purification and Quality Assessment in Paenibacillus Species
Journal of Food Protection
CTC staining
Flow cytometry
HistDenz treatment
Lysozyme treatment
Spore purification
title Optimization of Spore Purification and Quality Assessment in Paenibacillus Species
title_full Optimization of Spore Purification and Quality Assessment in Paenibacillus Species
title_fullStr Optimization of Spore Purification and Quality Assessment in Paenibacillus Species
title_full_unstemmed Optimization of Spore Purification and Quality Assessment in Paenibacillus Species
title_short Optimization of Spore Purification and Quality Assessment in Paenibacillus Species
title_sort optimization of spore purification and quality assessment in paenibacillus species
topic CTC staining
Flow cytometry
HistDenz treatment
Lysozyme treatment
Spore purification
url http://www.sciencedirect.com/science/article/pii/S0362028X25001474
work_keys_str_mv AT souichiroukawai optimizationofsporepurificationandqualityassessmentinpaenibacillusspecies
AT miyonakano optimizationofsporepurificationandqualityassessmentinpaenibacillusspecies