Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores
ABSTRACT 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 (Greenla...
Published in: | Applied and Environmental Microbiology |
---|---|
Main Authors: | , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
American Society for Microbiology
2006
|
Subjects: | |
Online Access: | http://dx.doi.org/10.1128/aem.00255-06 https://journals.asm.org/doi/pdf/10.1128/AEM.00255-06 |
id |
crasmicro:10.1128/aem.00255-06 |
---|---|
record_format |
openpolar |
spelling |
crasmicro:10.1128/aem.00255-06 2024-06-23T07:50:14+00:00 Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores Shafaat, Hannah S. Ponce, Adrian 2006 http://dx.doi.org/10.1128/aem.00255-06 https://journals.asm.org/doi/pdf/10.1128/AEM.00255-06 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Applied and Environmental Microbiology volume 72, issue 10, page 6808-6814 ISSN 0099-2240 1098-5336 journal-article 2006 crasmicro https://doi.org/10.1128/aem.00255-06 2024-05-27T13:00:05Z ABSTRACT 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 (Tb 3+ )-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/m 2 , 3,947 J/m 2 , and 1,322 J/m 2 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. Article in Journal/Newspaper Arctic Greenland Greenland Ice Sheet Project ice core Ice Sheet ASM Journals (American Society for Microbiology) Arctic Greenland Applied and Environmental Microbiology 72 10 6808 6814 |
institution |
Open Polar |
collection |
ASM Journals (American Society for Microbiology) |
op_collection_id |
crasmicro |
language |
English |
description |
ABSTRACT 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 (Tb 3+ )-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/m 2 , 3,947 J/m 2 , and 1,322 J/m 2 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 |
Article in Journal/Newspaper |
author |
Shafaat, Hannah S. Ponce, Adrian |
spellingShingle |
Shafaat, Hannah S. Ponce, Adrian Applications of a Rapid Endospore Viability Assay for Monitoring UV Inactivation and Characterizing Arctic Ice Cores |
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://dx.doi.org/10.1128/aem.00255-06 https://journals.asm.org/doi/pdf/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_source |
Applied and Environmental Microbiology volume 72, issue 10, page 6808-6814 ISSN 0099-2240 1098-5336 |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
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 |
_version_ |
1802641115608776704 |