A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products

IntroductionThe fundamental step in microbiological detection is the preparation of culture medium. The reliability and accuracy of microbiological assay heavily relies on this step. Currently in China, the most recognized standard method for such medium evaluation is ISO 11133-2014. However, this m...

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Main Authors: Wen Cui, Liang Zhao, Yuwen Shi, Wei Feng, Xiao Chen, Hui Liu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-02-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1493947/full
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author Wen Cui
Liang Zhao
Yuwen Shi
Wei Feng
Xiao Chen
Hui Liu
author_facet Wen Cui
Liang Zhao
Yuwen Shi
Wei Feng
Xiao Chen
Hui Liu
author_sort Wen Cui
collection DOAJ
description IntroductionThe fundamental step in microbiological detection is the preparation of culture medium. The reliability and accuracy of microbiological assay heavily relies on this step. Currently in China, the most recognized standard method for such medium evaluation is ISO 11133-2014. However, this method requires highly complicated biosafety management, detailed standard strains record-keeping and substantial working time.Methods and resultsBacterial tablet is considered to be a novel strategy for bacteria culture medium evaluation. The filter membrane as a carrier ensures uniform cell dispersion, forming a singular colony that can facilitate counting. We examined the viability and storage durability of vacuum freeze-dried bacterial tablets using a filter membrane as a carrier and utilized the results to evaluate culture medium. We found that the recipe GBSS (Glycerinum, Brain Heart Infusion, Sucrose, Sodium Glutamate) showed the highest survival rate for Escherichia coli in vacuum freeze-drying. As a qualified medium, the productivity of target bacterium should be greater than 0.7. A comparison of freeze-dried bacterial tablet method with ISO 11133-2014 quantitative method showed the sensitivity and specificity of this novel method were 94.1% and 88.9% respectively. The results suggested that vacuum freeze-dried bacterial tablet method had high conformity when compared to ISO 11133-2014 quantitative method (χ2 = 0.25, p > 0.05; Kappa = 0.75).DiscussionHence, vacuum freeze-drying method is an integral part of preservation of bacterial strains and the preparation of related biological products. In conclusion, we have developed a novel and effective disposable product for estimating efficiency of the culture medium.
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institution Kabale University
issn 1664-302X
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publishDate 2025-02-01
publisher Frontiers Media S.A.
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spelling doaj-art-a136eb318a9d434f824b9279bcc4e3c62025-02-10T06:48:44ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-02-011610.3389/fmicb.2025.14939471493947A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic productsWen Cui0Liang Zhao1Yuwen Shi2Wei Feng3Xiao Chen4Hui Liu5Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan, Shandong, ChinaDepartment of Anesthesiology, Children’s Hospital Affiliated to Shandong University and Jinan Children’s Hospital, Jinan, Shandong, ChinaJinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan, Shandong, ChinaDepartment of Clinical Laboratory, Yantai Yuhuangding Hospital, Yantai, ChinaJinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan, Shandong, ChinaJinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan, Shandong, ChinaIntroductionThe fundamental step in microbiological detection is the preparation of culture medium. The reliability and accuracy of microbiological assay heavily relies on this step. Currently in China, the most recognized standard method for such medium evaluation is ISO 11133-2014. However, this method requires highly complicated biosafety management, detailed standard strains record-keeping and substantial working time.Methods and resultsBacterial tablet is considered to be a novel strategy for bacteria culture medium evaluation. The filter membrane as a carrier ensures uniform cell dispersion, forming a singular colony that can facilitate counting. We examined the viability and storage durability of vacuum freeze-dried bacterial tablets using a filter membrane as a carrier and utilized the results to evaluate culture medium. We found that the recipe GBSS (Glycerinum, Brain Heart Infusion, Sucrose, Sodium Glutamate) showed the highest survival rate for Escherichia coli in vacuum freeze-drying. As a qualified medium, the productivity of target bacterium should be greater than 0.7. A comparison of freeze-dried bacterial tablet method with ISO 11133-2014 quantitative method showed the sensitivity and specificity of this novel method were 94.1% and 88.9% respectively. The results suggested that vacuum freeze-dried bacterial tablet method had high conformity when compared to ISO 11133-2014 quantitative method (χ2 = 0.25, p > 0.05; Kappa = 0.75).DiscussionHence, vacuum freeze-drying method is an integral part of preservation of bacterial strains and the preparation of related biological products. In conclusion, we have developed a novel and effective disposable product for estimating efficiency of the culture medium.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1493947/fullvacuum freeze-dryingculture mediumcryoprotectantbacterial tabletprobiotic
spellingShingle Wen Cui
Liang Zhao
Yuwen Shi
Wei Feng
Xiao Chen
Hui Liu
A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products
Frontiers in Microbiology
vacuum freeze-drying
culture medium
cryoprotectant
bacterial tablet
probiotic
title A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products
title_full A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products
title_fullStr A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products
title_full_unstemmed A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products
title_short A vacuum lyophilization and bacterial tablet-based method for culture medium evaluation and potential use in probiotic products
title_sort vacuum lyophilization and bacterial tablet based method for culture medium evaluation and potential use in probiotic products
topic vacuum freeze-drying
culture medium
cryoprotectant
bacterial tablet
probiotic
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1493947/full
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