Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria
Abstract Background Workers of agriculture and intensive life stock farming are exposed to highly contaminated workplaces. Bioaerosol exposures are suspected to trigger respiratory health effects of the workers. So far, risk evaluation of bioaerosols has been assessed through the infectivity of comp...
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doaj-31c0b8ac92814432b501c32b7e116d722021-08-15T11:16:29ZengBMCJournal of Occupational Medicine and Toxicology1745-66732021-08-0116111010.1186/s12995-021-00317-zApplication of impedance measurement to investigate in vitro inhalation toxicity of bacteriaStefanie Klar0Dierk-Christoph Poether1Jessica Reinert2Nicole Hüttig3Gunter Linsel4Udo Jäckel5Federal Institute for Occupational Safety and HealthFederal Institute for Occupational Safety and HealthFederal Institute for Occupational Safety and HealthFederal Institute for Occupational Safety and HealthFederal Institute for Occupational Safety and HealthFederal Institute for Occupational Safety and HealthAbstract Background Workers of agriculture and intensive life stock farming are exposed to highly contaminated workplaces. Bioaerosol exposures are suspected to trigger respiratory health effects of the workers. So far, risk evaluation of bioaerosols has been assessed through the infectivity of comprising biological agents that is classified in Europe by four risk groups according to the criteria of Directive 2000/54EC of the European Parliament. However, this directive additionally requires the risk assessment of allergenic and toxigenic effects without further elaboration. The aim of our study was to establish an in vitro screening system that is able to measure inhalative toxic effects of bacteria and their metabolites. Methods In this study, we analyzed three bacterial toxins and five culture supernatants of selected bacteria with known toxicity as model agents exposed to the lung epithelial cell line NuLi-1. We used electrical cell-substrate impedance sensing (ECIS) method to monitor real-time cell changes and the viability test Prestoblue™. Results We confirmed concentration dependent cytotoxic effects of the selected toxins in NuLi-1 cells over a period of up to 48 h. Each toxin resulted in a different but specific impedance profile over time according to their mode of action, whereas viability assay showed the metabolic activity of the cells at a chosen time point without revealing any information on their mode of action. Furthermore, dose-response-relationships were monitored. Tested model bacteria (Streptoccous pneumoniae, Acinetobacter radioresistens, Aerococcus viridans, Aeromonas hydrophila) reacted according to their expected toxicity except one bacterium (Enterococcus faecalis). The established assays revealed the concentration dependent onset and intensity of bacterial cytotoxicity and the viability of the cells at 24 h and 48 h exposure. Conclusion Impedance measurement and the viability assay Prestoblue™ in combination are suitable as sensitive screening methods to analyze toxic potential of bacteria and can therefor support the risk assessment of workplaces in terms of the directive 2000/54/EC.https://doi.org/10.1186/s12995-021-00317-zECISCytotoxicityBacteriaOccupational healthIn vitroNuLi-1 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Stefanie Klar Dierk-Christoph Poether Jessica Reinert Nicole Hüttig Gunter Linsel Udo Jäckel |
spellingShingle |
Stefanie Klar Dierk-Christoph Poether Jessica Reinert Nicole Hüttig Gunter Linsel Udo Jäckel Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria Journal of Occupational Medicine and Toxicology ECIS Cytotoxicity Bacteria Occupational health In vitro NuLi-1 |
author_facet |
Stefanie Klar Dierk-Christoph Poether Jessica Reinert Nicole Hüttig Gunter Linsel Udo Jäckel |
author_sort |
Stefanie Klar |
title |
Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria |
title_short |
Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria |
title_full |
Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria |
title_fullStr |
Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria |
title_full_unstemmed |
Application of impedance measurement to investigate in vitro inhalation toxicity of bacteria |
title_sort |
application of impedance measurement to investigate in vitro inhalation toxicity of bacteria |
publisher |
BMC |
series |
Journal of Occupational Medicine and Toxicology |
issn |
1745-6673 |
publishDate |
2021-08-01 |
description |
Abstract Background Workers of agriculture and intensive life stock farming are exposed to highly contaminated workplaces. Bioaerosol exposures are suspected to trigger respiratory health effects of the workers. So far, risk evaluation of bioaerosols has been assessed through the infectivity of comprising biological agents that is classified in Europe by four risk groups according to the criteria of Directive 2000/54EC of the European Parliament. However, this directive additionally requires the risk assessment of allergenic and toxigenic effects without further elaboration. The aim of our study was to establish an in vitro screening system that is able to measure inhalative toxic effects of bacteria and their metabolites. Methods In this study, we analyzed three bacterial toxins and five culture supernatants of selected bacteria with known toxicity as model agents exposed to the lung epithelial cell line NuLi-1. We used electrical cell-substrate impedance sensing (ECIS) method to monitor real-time cell changes and the viability test Prestoblue™. Results We confirmed concentration dependent cytotoxic effects of the selected toxins in NuLi-1 cells over a period of up to 48 h. Each toxin resulted in a different but specific impedance profile over time according to their mode of action, whereas viability assay showed the metabolic activity of the cells at a chosen time point without revealing any information on their mode of action. Furthermore, dose-response-relationships were monitored. Tested model bacteria (Streptoccous pneumoniae, Acinetobacter radioresistens, Aerococcus viridans, Aeromonas hydrophila) reacted according to their expected toxicity except one bacterium (Enterococcus faecalis). The established assays revealed the concentration dependent onset and intensity of bacterial cytotoxicity and the viability of the cells at 24 h and 48 h exposure. Conclusion Impedance measurement and the viability assay Prestoblue™ in combination are suitable as sensitive screening methods to analyze toxic potential of bacteria and can therefor support the risk assessment of workplaces in terms of the directive 2000/54/EC. |
topic |
ECIS Cytotoxicity Bacteria Occupational health In vitro NuLi-1 |
url |
https://doi.org/10.1186/s12995-021-00317-z |
work_keys_str_mv |
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