Metabolic indicators for assessing bacterial viability in hygiene sampling using cells in suspension and swabbed biofilm

Johanna Maukonen, Tiina Mattila-Sandholm, Gun Wirtanen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

13 Citations (Scopus)

Abstract

In the present study various combinations of metabolic indicators were used in viability studies of foodborne spoilage and pathogenic organisms. Various metabolic stains were tested in pure culture suspensions of Pseudomonas fragi and Listeria monocytogenes containing only viable cells, both viable and dead cells or only dead cells. In addition, foodborne monospecies biofilms of L. monocytogenes and P. fragi were studied. The biofilms were grown on stainless steel (AISI 304, 2B) surfaces. The results showed that both the LIVE/DEAD Bac Light Viability Kit (LIVE/DEAD) and the 5-cyano-2,3-ditolyl tetrazolium chloride (CTC)-4′,6-diamidino-2-phenylindole (DAPI) staining procedures offer a rapid, easy and reliable method for metabolic investigation of bacteria in suspension and swabbed biofilm bacteria. Assessment of replicate samples using luciferin–luciferase-based ATP measurement and conventional cultivation corroborated the results of swabbed biofilm bacteria obtained by staining. LIVE/DEAD staining cannot be used for direct staining of biofilms on surfaces, due to interference between the stain and polysaccharides of the biofilm matrix and slime. Staining by CTC-DAPI is suitable for studying viability of cells both in suspension and in biofilms attached to surfaces. The other staining procedures tested were not satisfactory or only slightly satisfactory in distinguishing between viable and dead cells.
Original languageEnglish
Pages (from-to)225 - 233
Number of pages9
JournalLWT - Food Science and Technology
Volume33
Issue number3
DOIs
Publication statusPublished - 2000
MoE publication typeA1 Journal article-refereed

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Microbial Viability
Biofilms
Hygiene
hygiene
biofilm
cell suspension culture
Suspensions
viability
Staining and Labeling
Pseudomonas fragi
tetrazolium
sampling
Listeria monocytogenes
Bacteria
dyes
bacteria
chlorides
Coloring Agents
cells
Stainless Steel

Cite this

Maukonen, Johanna ; Mattila-Sandholm, Tiina ; Wirtanen, Gun. / Metabolic indicators for assessing bacterial viability in hygiene sampling using cells in suspension and swabbed biofilm. In: LWT - Food Science and Technology. 2000 ; Vol. 33, No. 3. pp. 225 - 233.
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Metabolic indicators for assessing bacterial viability in hygiene sampling using cells in suspension and swabbed biofilm. / Maukonen, Johanna; Mattila-Sandholm, Tiina; Wirtanen, Gun (Corresponding Author).

In: LWT - Food Science and Technology, Vol. 33, No. 3, 2000, p. 225 - 233.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Metabolic indicators for assessing bacterial viability in hygiene sampling using cells in suspension and swabbed biofilm

AU - Maukonen, Johanna

AU - Mattila-Sandholm, Tiina

AU - Wirtanen, Gun

PY - 2000

Y1 - 2000

N2 - In the present study various combinations of metabolic indicators were used in viability studies of foodborne spoilage and pathogenic organisms. Various metabolic stains were tested in pure culture suspensions of Pseudomonas fragi and Listeria monocytogenes containing only viable cells, both viable and dead cells or only dead cells. In addition, foodborne monospecies biofilms of L. monocytogenes and P. fragi were studied. The biofilms were grown on stainless steel (AISI 304, 2B) surfaces. The results showed that both the LIVE/DEAD Bac Light Viability Kit (LIVE/DEAD) and the 5-cyano-2,3-ditolyl tetrazolium chloride (CTC)-4′,6-diamidino-2-phenylindole (DAPI) staining procedures offer a rapid, easy and reliable method for metabolic investigation of bacteria in suspension and swabbed biofilm bacteria. Assessment of replicate samples using luciferin–luciferase-based ATP measurement and conventional cultivation corroborated the results of swabbed biofilm bacteria obtained by staining. LIVE/DEAD staining cannot be used for direct staining of biofilms on surfaces, due to interference between the stain and polysaccharides of the biofilm matrix and slime. Staining by CTC-DAPI is suitable for studying viability of cells both in suspension and in biofilms attached to surfaces. The other staining procedures tested were not satisfactory or only slightly satisfactory in distinguishing between viable and dead cells.

AB - In the present study various combinations of metabolic indicators were used in viability studies of foodborne spoilage and pathogenic organisms. Various metabolic stains were tested in pure culture suspensions of Pseudomonas fragi and Listeria monocytogenes containing only viable cells, both viable and dead cells or only dead cells. In addition, foodborne monospecies biofilms of L. monocytogenes and P. fragi were studied. The biofilms were grown on stainless steel (AISI 304, 2B) surfaces. The results showed that both the LIVE/DEAD Bac Light Viability Kit (LIVE/DEAD) and the 5-cyano-2,3-ditolyl tetrazolium chloride (CTC)-4′,6-diamidino-2-phenylindole (DAPI) staining procedures offer a rapid, easy and reliable method for metabolic investigation of bacteria in suspension and swabbed biofilm bacteria. Assessment of replicate samples using luciferin–luciferase-based ATP measurement and conventional cultivation corroborated the results of swabbed biofilm bacteria obtained by staining. LIVE/DEAD staining cannot be used for direct staining of biofilms on surfaces, due to interference between the stain and polysaccharides of the biofilm matrix and slime. Staining by CTC-DAPI is suitable for studying viability of cells both in suspension and in biofilms attached to surfaces. The other staining procedures tested were not satisfactory or only slightly satisfactory in distinguishing between viable and dead cells.

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DO - 10.1006/fstl.2000.0650

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