Optimization of Spore Purification and Quality Assessment in Paenibacillus Species

IF 2.8 4区农林科学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of food protection Pub Date : 2025-08-06 DOI:10.1016/j.jfp.2025.100595

Souichirou Kawai, Miyo Nakano

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引用次数: 0

Abstract

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 21743^T and JCM 13339, P. terrae JCM 11466^T, and P. polymyxa JCM 2507^T) 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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芽孢杆菌孢子纯化及质量评价的优化。

为了提高食品安全研究中应用的芽孢杆菌孢子制剂的一致性和质量，在不同的产孢条件下（固体琼脂培养基和液体培养基），研究了溶菌酶处理和HistDenz梯度离心两种常用的纯化方法。对4株芽孢杆菌（P. odorifer JCM 21743T和JCM 13339， P. terrae JCM 11466T和P. polymyxa JCM 2507T）的孢子进行纯化，利用5-氰基-2,3-二醇四氯化铵（CTC）染色法评价孢子的代谢活性，并通过生长曲线分析孢子的萌发能力。CTC染色证实在所有纯化的样品中完全没有代谢活性的营养细胞。生长试验表明，溶菌酶处理降低了某些菌株的孢子活力，而HistDenz纯化更可靠地保留了萌发潜力。此外，与某些菌株在液体培养基中产生的孢子相比，在固体培养基中产生的孢子表现出更好的生长性能-证明了更高的峰值浊度和更短的倍增时间。这些结果表明，纯化技术和产孢环境对芽孢杆菌孢子的生理品质有显著影响。因此，选择菌株特异性制备方案对于准确评估孢子活力和确保其在下游检测或安全性评估中的有效应用至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of food protection 工程技术-生物工程与应用微生物

CiteScore

4.20

自引率

5.00%

发文量

296

审稿时长

2.5 months

期刊介绍： The Journal of Food Protection® (JFP) is an international, monthly scientific journal in the English language published by the International Association for Food Protection (IAFP). JFP publishes research and review articles on all aspects of food protection and safety. Major emphases of JFP are placed on studies dealing with: Tracking, detecting (including traditional, molecular, and real-time), inactivating, and controlling food-related hazards, including microorganisms (including antibiotic resistance), microbial (mycotoxins, seafood toxins) and non-microbial toxins (heavy metals, pesticides, veterinary drug residues, migrants from food packaging, and processing contaminants), allergens and pests (insects, rodents) in human food, pet food and animal feed throughout the food chain; Microbiological food quality and traditional/novel methods to assay microbiological food quality; Prevention of food-related hazards and food spoilage through food preservatives and thermal/non-thermal processes, including process validation; Food fermentations and food-related probiotics; Safe food handling practices during pre-harvest, harvest, post-harvest, distribution and consumption, including food safety education for retailers, foodservice, and consumers; Risk assessments for food-related hazards; Economic impact of food-related hazards, foodborne illness, food loss, food spoilage, and adulterated foods; Food fraud, food authentication, food defense, and foodborne disease outbreak investigations.