دورية أكاديمية
أعرض في EDS

Effect of cryopreservation medium conditions on growth and isolation of gut anaerobes from human faecal samples.

التفاصيل البيبلوغرافية
العنوان: Effect of cryopreservation medium conditions on growth and isolation of gut anaerobes from human faecal samples.
المؤلفون: Biclot A; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium., Huys GRB; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium., Bacigalupe R; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium., D'hoe K; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium.; Research Group of Microbiology, Department of Bioengineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium., Vandeputte D; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium.; Research Group of Microbiology, Department of Bioengineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium.; Present address: Meinig School of Biomedical Engineering, Cornell, USA., Falony G; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium., Tito RY; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium.; Center for Microbiology, VIB, Leuven, Belgium., Raes J; Laboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute, KU Leuven, Leuven, Belgium. jeroen.raes@kuleuven.be.; Center for Microbiology, VIB, Leuven, Belgium. jeroen.raes@kuleuven.be.
المصدر: Microbiome [Microbiome] 2022 May 30; Vol. 10 (1), pp. 80. Date of Electronic Publication: 2022 May 30.
نوع المنشور: Journal Article; Video-Audio Media; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: BioMed Central Country of Publication: England NLM ID: 101615147 Publication Model: Electronic Cited Medium: Internet ISSN: 2049-2618 (Electronic) Linking ISSN: 20492618 NLM ISO Abbreviation: Microbiome Subsets: MEDLINE
أسماء مطبوعة: Original Publication: London: BioMed Central, 2013-
مواضيع طبية MeSH: Cryopreservation* , Dimethyl Sulfoxide*, Culture Media ; Feces/microbiology ; Humans ; RNA, Ribosomal, 16S/genetics
مستخلص: Background: Novel strategies for anaerobic bacterial isolations from human faecal samples and various initiatives to generate culture collections of gut-derived bacteria have instigated considerable interest for the development of novel microbiota-based treatments. Early in the process of building a culture collection, optimal faecal sample preservation is essential to safeguard the viability of the broadest taxonomic diversity range possible. In contrast to the much more established faecal storage conditions for meta-omics applications, the impact of stool sample preservation conditions on bacterial growth recovery and isolation remains largely unexplored. In this study, aliquoted faecal samples from eleven healthy human volunteers selected based on a range of physicochemical and microbiological gradients were cryopreserved at - 80 °C either without the addition of any medium (dry condition) or in different Cary-Blair medium conditions with or without a cryoprotectant, i.e. 20% (v/v) glycerol or 5% (v/v) DMSO. Faecal aliquots were subjected to bulk 16S rRNA gene sequencing as well as dilution plating on modified Gifu Anaerobic Medium after preservation for culturable fraction profiling and generation of bacterial culture collections.
Results: Analyses of compositional variation showed that cryopreservation medium conditions affected quantitative recovery but not the overall community composition of cultured fractions. Post-preservation sample dilution and richness of the uncultured source samples were the major drivers of the cultured fraction richness at genus level. However, preservation conditions differentially affected recovery of specific genera. Presence-absence analysis indicated that twenty-two of the 45 most abundant common genera (>0.01% abundance, dilution 10 -4 ) were recovered in cultured fractions from all preservation conditions, while nine genera were only detected in fractions from a single preservation condition. Overall, the highest number of common genera (i.e. 35/45) in cultured fractions were recovered from sample aliquots preserved without medium and in the presence of Cary-Blair medium containing 5% (v/v) DMSO. Also, in the culture collection generated from the cultured fractions, these two preservation conditions yielded the highest species richness (72 and 66, respectively).
Conclusion: Our results demonstrate that preservation methods partly determine richness and taxonomic diversity of gut anaerobes recovered from faecal samples. Complementing the current standard practice of cryopreserving stool samples in dry conditions with other preservation conditions, such as Cary-Blair medium with DMSO, could increase the species diversity of gut-associated culture collections. Video abstract.
(© 2022. The Author(s).)
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المشرفين على المادة: 0 (Culture Media)
0 (RNA, Ribosomal, 16S)
YOW8V9698H (Dimethyl Sulfoxide)
تواريخ الأحداث: Date Created: 20220601 Date Completed: 20220603 Latest Revision: 20240830
رمز التحديث: 20240830
مُعرف محوري في PubMed: PMC9150342
DOI: 10.1186/s40168-022-01267-2
PMID: 35644616
قاعدة البيانات: MEDLINE
الوصف
تدمد:2049-2618
DOI:10.1186/s40168-022-01267-2