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Different methods to quantify Listeria monocytogenes biofilms cells showed different profile in their viability.

Authors :
Winkelströter LK
De Martinis EC
Source :
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Braz J Microbiol] 2015 Mar 01; Vol. 46 (1), pp. 231-5. Date of Electronic Publication: 2015 Mar 01 (Print Publication: 2015).
Publication Year :
2015

Abstract

Listeria monocytogenes is a foodborne pathogen able to adhere and to form biofilms in several materials commonly present in food processing plants. The aim of this study was to evaluate the resistance of Listeria monocytogenes attached to abiotic surface, after treatment with sanitizers, by culture method, microscopy and Quantitative Real Time Polymerase Chain Reaction (qPCR). Biofilms of L. monocytogenes were obtained in stainless steel coupons immersed in Brain Heart Infusion Broth, under agitation at 37 °C for 24 h. The methods selected for this study were based on plate count, microscopic count with the aid of viability dyes (CTC-DAPI), and qPCR. Results of culture method showed that peroxyacetic acid was efficient to kill sessile L. monocytogenes populations, while sodium hypochlorite was only partially effective to kill attached L. monocytogenes (p < 0.05). When, viability dyes (CTC/DAPI) combined with fluorescence microscopy and qPCR were used and lower counts were found after treatments (p < 0.05). Selective quantification of viable cells of L. monocytogenes by qPCR using EMA revelead that the pre-treatment with EMA was not appropriate since it also inhibited amplification of DNA from live cells by ca. 2 log. Thus, the use of CTC counts was the best method to count viable cells in biofilms.

Details

Language :
English
ISSN :
1678-4405
Volume :
46
Issue :
1
Database :
MEDLINE
Journal :
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
Publication Type :
Academic Journal
Accession number :
26221112
Full Text :
https://doi.org/10.1590/S1517-838220131071