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Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples

Culture-independent sequence data from various environmental samples have revealed an immense microbial diversity of environmental, clinical, and industrial importance that has not yet been cultured. Cultivation is imperative to validate findings emerging from cultivation-independent molecular data...

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Detalles Bibliográficos
Autores principales: Yadav, Krishna K., Nimonkar, Yogesh, Poddar, Bhagyashri J., Kovale, Lochana, Sagar, Isita, Shouche, Yogesh, Purohit, Hemant J., Khardenavis, Anshuman A., Green, Stefan J., Prakash, Om
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9248899/
https://www.ncbi.nlm.nih.gov/pubmed/35467387
http://dx.doi.org/10.1128/spectrum.00007-22
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author Yadav, Krishna K.
Nimonkar, Yogesh
Poddar, Bhagyashri J.
Kovale, Lochana
Sagar, Isita
Shouche, Yogesh
Purohit, Hemant J.
Khardenavis, Anshuman A.
Green, Stefan J.
Prakash, Om
author_facet Yadav, Krishna K.
Nimonkar, Yogesh
Poddar, Bhagyashri J.
Kovale, Lochana
Sagar, Isita
Shouche, Yogesh
Purohit, Hemant J.
Khardenavis, Anshuman A.
Green, Stefan J.
Prakash, Om
author_sort Yadav, Krishna K.
collection PubMed
description Culture-independent sequence data from various environmental samples have revealed an immense microbial diversity of environmental, clinical, and industrial importance that has not yet been cultured. Cultivation is imperative to validate findings emerging from cultivation-independent molecular data and exploit the isolated organisms for biotechnological purposes. Efforts have been made to boost the cultivability of microbes from environmental samples by use of a range of techniques and instrumentation. The manuscript presents a novel yet simple and innovative approach to improving the cultivability of natural microorganisms without sophisticated instrumentation. By employing gradient centrifugation combined with serial dilution (“two-dimensional cell separation”), significantly higher numbers of genera (>2-fold higher) and species (>3-fold higher) were isolated from environmental samples, including soil, anaerobic sludge, and landfill leachate, than from using serial dilution alone. This simple and robust protocol can be modified for any environment and culture medium and provides access to untapped microbial diversity. IMPORTANCE In the manuscript, we have developed a novel yet simple and innovative approach to improving the cultivability of natural microorganisms without sophisticated instrumentation. The method used gradient centrifugation combined with serial dilution (two-dimensional cell separation) to improve taxum recovery from samples. This simple and robust protocol can be modified for any environment and culture medium and provides access to untapped microbial diversity. This approach can be incorporated with less labor and complexity in laboratories with minimal instrumentation. As cultivation is a workflow that is well suited to lower-resource microbiology labs, we believe improvements in cultivability can increase opportunities for scientific collaborations between low-resource labs and groups focused on high-resource cultivation-independent methodologies.
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spelling pubmed-92488992022-07-02 Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples Yadav, Krishna K. Nimonkar, Yogesh Poddar, Bhagyashri J. Kovale, Lochana Sagar, Isita Shouche, Yogesh Purohit, Hemant J. Khardenavis, Anshuman A. Green, Stefan J. Prakash, Om Microbiol Spectr Research Article Culture-independent sequence data from various environmental samples have revealed an immense microbial diversity of environmental, clinical, and industrial importance that has not yet been cultured. Cultivation is imperative to validate findings emerging from cultivation-independent molecular data and exploit the isolated organisms for biotechnological purposes. Efforts have been made to boost the cultivability of microbes from environmental samples by use of a range of techniques and instrumentation. The manuscript presents a novel yet simple and innovative approach to improving the cultivability of natural microorganisms without sophisticated instrumentation. By employing gradient centrifugation combined with serial dilution (“two-dimensional cell separation”), significantly higher numbers of genera (>2-fold higher) and species (>3-fold higher) were isolated from environmental samples, including soil, anaerobic sludge, and landfill leachate, than from using serial dilution alone. This simple and robust protocol can be modified for any environment and culture medium and provides access to untapped microbial diversity. IMPORTANCE In the manuscript, we have developed a novel yet simple and innovative approach to improving the cultivability of natural microorganisms without sophisticated instrumentation. The method used gradient centrifugation combined with serial dilution (two-dimensional cell separation) to improve taxum recovery from samples. This simple and robust protocol can be modified for any environment and culture medium and provides access to untapped microbial diversity. This approach can be incorporated with less labor and complexity in laboratories with minimal instrumentation. As cultivation is a workflow that is well suited to lower-resource microbiology labs, we believe improvements in cultivability can increase opportunities for scientific collaborations between low-resource labs and groups focused on high-resource cultivation-independent methodologies. American Society for Microbiology 2022-04-25 /pmc/articles/PMC9248899/ /pubmed/35467387 http://dx.doi.org/10.1128/spectrum.00007-22 Text en Copyright © 2022 Yadav et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Yadav, Krishna K.
Nimonkar, Yogesh
Poddar, Bhagyashri J.
Kovale, Lochana
Sagar, Isita
Shouche, Yogesh
Purohit, Hemant J.
Khardenavis, Anshuman A.
Green, Stefan J.
Prakash, Om
Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples
title Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples
title_full Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples
title_fullStr Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples
title_full_unstemmed Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples
title_short Two-Dimensional Cell Separation: a High-Throughput Approach to Enhance the Culturability of Bacterial Cells from Environmental Samples
title_sort two-dimensional cell separation: a high-throughput approach to enhance the culturability of bacterial cells from environmental samples
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9248899/
https://www.ncbi.nlm.nih.gov/pubmed/35467387
http://dx.doi.org/10.1128/spectrum.00007-22
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