Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments
Bacterial endospores are formed in genera such as Bacillus and Clostridium in response to adverse environmental changes. Endospores have remarkable resistance to various extreme conditions and can remain dormant for extended periods of time. Clostridium spores are of particular interest due to their...
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ftdatacite:10.7907/8bbd-7y02 2023-05-15T16:30:33+02:00 Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments Yang, Wanwan 2010 PDF https://dx.doi.org/10.7907/8bbd-7y02 https://resolver.caltech.edu/CaltechTHESIS:01072010-132355433 en eng California Institute of Technology No commercial reproduction, distribution, display or performance rights in this work are provided. Bacterial Spore Extreme Environments Clostridium Fluorescence Detection Method Environmental Science and Engineering Thesis Text Dissertation thesis 2010 ftdatacite https://doi.org/10.7907/8bbd-7y02 2021-11-05T12:55:41Z Bacterial endospores are formed in genera such as Bacillus and Clostridium in response to adverse environmental changes. Endospores have remarkable resistance to various extreme conditions and can remain dormant for extended periods of time. Clostridium spores are of particular interest due to their significant importance in several industries, such as food processing, wastewater treatment, pharmaceuticals, and health care. They are also the ideal candidates to study Panspermia and potential extraterrestrial life. However, to date, most endospore research has been conducted on Bacillus, and study of the anaerobic spore former, Clostridium, is not adequate. In this study, we have developed a general protocol to produce and purify Clostridium spores. Spectroscopy and microscopy based Endospore Viability Assay (Spectro EVA and Micro EVA) were developed and validated to assess the viability of Clostridium spores. Germinability was used as an indicator for spore viability. The basic principle of the two EVAs is to measure the release of a unique biomarker, dipicolinic acid (DPA), via germination as a proxy for endospore viability. In particular, a luminescence time-gated microscopy technique (Micro EVA) has been developed to enumerate germinable Clostridium endospores within an hour. Micro EVA is based on energy transfer from DPA to terbium ions doped in a solid matrix upon UV excitation. The distinctive emission and millisecond lifetime enables time-resolved imaging to achieve single endospore sensitivity. Comparing to traditional CFU cultivation, EVA probes the early stage of germination, resulting in a much faster detection rate (within 60 minutes) than CFU measurement (more than 3 days incubation). Micro EVA has also been successfully applied to quantify Clostridium spores in an extreme cold biosphere, Greenland ice core, and a hyper-arid biosphere, Atacama Desert, two Mars analogs on earth. The development of EVA provides a faster way to assess viability of Clostridium spores, which has significant importance in various industries. It also enables the determination of the limit and longevity of life, and provides insight on the search of extinct or extant life on Mars and other celestial bodies. Thesis Greenland Greenland ice core ice core DataCite Metadata Store (German National Library of Science and Technology) Greenland |
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collection |
DataCite Metadata Store (German National Library of Science and Technology) |
op_collection_id |
ftdatacite |
language |
English |
topic |
Bacterial Spore Extreme Environments Clostridium Fluorescence Detection Method Environmental Science and Engineering |
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Bacterial Spore Extreme Environments Clostridium Fluorescence Detection Method Environmental Science and Engineering Yang, Wanwan Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments |
topic_facet |
Bacterial Spore Extreme Environments Clostridium Fluorescence Detection Method Environmental Science and Engineering |
description |
Bacterial endospores are formed in genera such as Bacillus and Clostridium in response to adverse environmental changes. Endospores have remarkable resistance to various extreme conditions and can remain dormant for extended periods of time. Clostridium spores are of particular interest due to their significant importance in several industries, such as food processing, wastewater treatment, pharmaceuticals, and health care. They are also the ideal candidates to study Panspermia and potential extraterrestrial life. However, to date, most endospore research has been conducted on Bacillus, and study of the anaerobic spore former, Clostridium, is not adequate. In this study, we have developed a general protocol to produce and purify Clostridium spores. Spectroscopy and microscopy based Endospore Viability Assay (Spectro EVA and Micro EVA) were developed and validated to assess the viability of Clostridium spores. Germinability was used as an indicator for spore viability. The basic principle of the two EVAs is to measure the release of a unique biomarker, dipicolinic acid (DPA), via germination as a proxy for endospore viability. In particular, a luminescence time-gated microscopy technique (Micro EVA) has been developed to enumerate germinable Clostridium endospores within an hour. Micro EVA is based on energy transfer from DPA to terbium ions doped in a solid matrix upon UV excitation. The distinctive emission and millisecond lifetime enables time-resolved imaging to achieve single endospore sensitivity. Comparing to traditional CFU cultivation, EVA probes the early stage of germination, resulting in a much faster detection rate (within 60 minutes) than CFU measurement (more than 3 days incubation). Micro EVA has also been successfully applied to quantify Clostridium spores in an extreme cold biosphere, Greenland ice core, and a hyper-arid biosphere, Atacama Desert, two Mars analogs on earth. The development of EVA provides a faster way to assess viability of Clostridium spores, which has significant importance in various industries. It also enables the determination of the limit and longevity of life, and provides insight on the search of extinct or extant life on Mars and other celestial bodies. |
format |
Thesis |
author |
Yang, Wanwan |
author_facet |
Yang, Wanwan |
author_sort |
Yang, Wanwan |
title |
Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments |
title_short |
Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments |
title_full |
Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments |
title_fullStr |
Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments |
title_full_unstemmed |
Fast Viability Assessment of Clostridium Spores: Survival in Extreme Environments |
title_sort |
fast viability assessment of clostridium spores: survival in extreme environments |
publisher |
California Institute of Technology |
publishDate |
2010 |
url |
https://dx.doi.org/10.7907/8bbd-7y02 https://resolver.caltech.edu/CaltechTHESIS:01072010-132355433 |
geographic |
Greenland |
geographic_facet |
Greenland |
genre |
Greenland Greenland ice core ice core |
genre_facet |
Greenland Greenland ice core ice core |
op_rights |
No commercial reproduction, distribution, display or performance rights in this work are provided. |
op_doi |
https://doi.org/10.7907/8bbd-7y02 |
_version_ |
1766020275372032000 |