Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores†
We have developed a rapid endospore viability assay (EVA) in which endospore germination serves as an indicator for viability and applied it to (i) monitor UV inactivation of endospores as a function of dose and (ii) determine the proportion of viable endospores in arctic ice cores (Greenland Ice Sh...
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ftpubmed:oai:pubmedcentral.nih.gov:1610317 2023-05-15T15:00:00+02:00 Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† Shafaat, Hannah S. Ponce, Adrian 2006-10 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1610317 http://www.ncbi.nlm.nih.gov/pubmed/17021233 https://doi.org/10.1128/AEM.00255-06 en eng American Society for Microbiology http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1610317 http://www.ncbi.nlm.nih.gov/pubmed/17021233 http://dx.doi.org/10.1128/AEM.00255-06 Copyright © 2006, American Society for Microbiology Methods Text 2006 ftpubmed https://doi.org/10.1128/AEM.00255-06 2013-08-31T08:48:36Z We have developed a rapid endospore viability assay (EVA) in which endospore germination serves as an indicator for viability and applied it to (i) monitor UV inactivation of endospores as a function of dose and (ii) determine the proportion of viable endospores in arctic ice cores (Greenland Ice Sheet Project 2 [GISP2] cores; 94 m). EVA is based on the detection of dipicolinic acid (DPA), which is released from endospores during germination. DPA concentrations were determined using the terbium ion (Tb3+)-DPA luminescence assay, and germination was induced by l-alanine addition. The concentrations of germinable endospores were determined by comparison to a standard curve. Parallel EVA and phase-contrast microscopy experiments to determine the percentage of germinable spores yielded comparable results (54.3% ± 3.8% and 48.9% ± 4.5%, respectively), while only 27.8% ± 7.6% of spores produced CFU. EVA was applied to monitor the inactivation of spore suspensions as a function of UV dose, yielding reproducible correlations between EVA and CFU inactivation data. The 90% inactivation doses were 2,773 J/m2, 3,947 J/m2, and 1,322 J/m2 for EVA, phase-contrast microscopy, and CFU reduction, respectively. Finally, EVA was applied to quantify germinable and total endospore concentrations in two GISP2 ice cores. The first ice core contained 295 ± 19 germinable spores/ml and 369 ± 36 total spores/ml (i.e., the percentage of germinable endospores was 79.9% ± 9.3%), and the second core contained 131 ± 4 germinable spores/ml and 162 ± 17 total spores/ml (i.e., the percentage of germinable endospores was 80.9% ± 8.8%), whereas only 2 CFU/ml were detected by culturing. Text Arctic Greenland Greenland Ice Sheet Project ice core Ice Sheet PubMed Central (PMC) Arctic Greenland Applied and Environmental Microbiology 72 10 6808 6814 |
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Methods Shafaat, Hannah S. Ponce, Adrian Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† |
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Methods |
description |
We have developed a rapid endospore viability assay (EVA) in which endospore germination serves as an indicator for viability and applied it to (i) monitor UV inactivation of endospores as a function of dose and (ii) determine the proportion of viable endospores in arctic ice cores (Greenland Ice Sheet Project 2 [GISP2] cores; 94 m). EVA is based on the detection of dipicolinic acid (DPA), which is released from endospores during germination. DPA concentrations were determined using the terbium ion (Tb3+)-DPA luminescence assay, and germination was induced by l-alanine addition. The concentrations of germinable endospores were determined by comparison to a standard curve. Parallel EVA and phase-contrast microscopy experiments to determine the percentage of germinable spores yielded comparable results (54.3% ± 3.8% and 48.9% ± 4.5%, respectively), while only 27.8% ± 7.6% of spores produced CFU. EVA was applied to monitor the inactivation of spore suspensions as a function of UV dose, yielding reproducible correlations between EVA and CFU inactivation data. The 90% inactivation doses were 2,773 J/m2, 3,947 J/m2, and 1,322 J/m2 for EVA, phase-contrast microscopy, and CFU reduction, respectively. Finally, EVA was applied to quantify germinable and total endospore concentrations in two GISP2 ice cores. The first ice core contained 295 ± 19 germinable spores/ml and 369 ± 36 total spores/ml (i.e., the percentage of germinable endospores was 79.9% ± 9.3%), and the second core contained 131 ± 4 germinable spores/ml and 162 ± 17 total spores/ml (i.e., the percentage of germinable endospores was 80.9% ± 8.8%), whereas only 2 CFU/ml were detected by culturing. |
format |
Text |
author |
Shafaat, Hannah S. Ponce, Adrian |
author_facet |
Shafaat, Hannah S. Ponce, Adrian |
author_sort |
Shafaat, Hannah S. |
title |
Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† |
title_short |
Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† |
title_full |
Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† |
title_fullStr |
Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† |
title_full_unstemmed |
Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores† |
title_sort |
applications of a rapid endospore viability assay for monitoring uv inactivation and characterizing arctic ice cores† |
publisher |
American Society for Microbiology |
publishDate |
2006 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1610317 http://www.ncbi.nlm.nih.gov/pubmed/17021233 https://doi.org/10.1128/AEM.00255-06 |
geographic |
Arctic Greenland |
geographic_facet |
Arctic Greenland |
genre |
Arctic Greenland Greenland Ice Sheet Project ice core Ice Sheet |
genre_facet |
Arctic Greenland Greenland Ice Sheet Project ice core Ice Sheet |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1610317 http://www.ncbi.nlm.nih.gov/pubmed/17021233 http://dx.doi.org/10.1128/AEM.00255-06 |
op_rights |
Copyright © 2006, American Society for Microbiology |
op_doi |
https://doi.org/10.1128/AEM.00255-06 |
container_title |
Applied and Environmental Microbiology |
container_volume |
72 |
container_issue |
10 |
container_start_page |
6808 |
op_container_end_page |
6814 |
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1766332110325415936 |