Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR
Brochothrix thermosphacta, a Cram-positive bacterium, is considered as the predominant spoilage microbiota of modified atmosphere packing (MAP) shrimp and fish. Traditional methods currently used to detect B. thermosphacta in foods are time-consuming and labour-intensive. The aim of this study was t...
Published in: | Food Microbiology |
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Format: | Article in Journal/Newspaper |
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Online Access: | https://hal.archives-ouvertes.fr/hal-01004204 https://doi.org/10.1016/j.fm.2011.09.012 |
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ftccsdartic:oai:HAL:hal-01004204v1 2023-05-15T17:54:32+02:00 Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR Mamlouk, Kelthoum, Mace, Sabrina, Guilbaud, Morgan, JAFFRES, Emmanuel, Ferchichi, Mounir, Prévost, Herve, Pilet, Marie-France, Dousset, Xavier Oniris, Secalim UMR1014 LUNAM Université Nantes Angers Le Mans MICrobiologie de l'ALImentation au Service de la Santé (MICALIS) Institut National de la Recherche Agronomique (INRA)-AgroParisTech SECurité des ALIments et Microbiologie Institut National de la Recherche Agronomique (INRA) Region Pays de la Loire, France 2012 https://hal.archives-ouvertes.fr/hal-01004204 https://doi.org/10.1016/j.fm.2011.09.012 en eng HAL CCSD Elsevier info:eu-repo/semantics/altIdentifier/doi/10.1016/j.fm.2011.09.012 hal-01004204 https://hal.archives-ouvertes.fr/hal-01004204 doi:10.1016/j.fm.2011.09.012 PRODINRA: 215388 WOS: 000300739900023 ISSN: 0740-0020 EISSN: 1095-9998 Food Microbiology https://hal.archives-ouvertes.fr/hal-01004204 Food Microbiology, Elsevier, 2012, 30 (1), pp.173 - 179. ⟨10.1016/j.fm.2011.09.012⟩ RAPQUANTITATIVE DETECTION STAA medium Seafood Spoilage Brochothrix thermosphacta Real-time PCR 16S rRNA COLD-SMOKED SALMON LISTERIA-MONOCYTOGENES PROPIDIUM MONOAZIDE PANDALUS-BOREALIS MICROBIAL ECOLOGY MEAT FOOD CARNOBACTERIUM LACTOCOCCUS [SDV.SA]Life Sciences [q-bio]/Agricultural sciences info:eu-repo/semantics/article Journal articles 2012 ftccsdartic https://doi.org/10.1016/j.fm.2011.09.012 2021-11-28T01:19:32Z Brochothrix thermosphacta, a Cram-positive bacterium, is considered as the predominant spoilage microbiota of modified atmosphere packing (MAP) shrimp and fish. Traditional methods currently used to detect B. thermosphacta in foods are time-consuming and labour-intensive. The aim of this study was to develop a real-time PCR quantification method combined with a propidium monoazide (PMA) sample treatment step to monitor the population of B. thermosphacta in cooked shrimp and salmon. The specificity of the two primers M0405 and M0404 used to amplify a 70 bp fragment of the 16S rRNA gene was demonstrated by using purified DNA from 30 strains, among 21 bacterial species including 22 reference strains. Using these primers for real-time PCR and in pure culture, a good correlation was obtained between real-time PCR and the conventional plating method. Quantification was linear over 7-log units using artificially inoculated samples. The method performed successfully when tested on naturally contaminated cooked shrimp and fresh salmon, with a minimum threshold of 1.9 x 10(2) CFU/g for accurate quantification of B. the rmosphacta. The correlation between the B. thermosphacta counts obtained by real-time PCR and plate counts on naturally contaminated shrimp and salmon was high (R-2=0.895). Thus, this study presents a rapid tool for producing reliable quantitative data on B. thermosphacta in cooked shrimp and fresh salmon. (C) 2011 Elsevier Ltd. All rights reserved. Article in Journal/Newspaper Pandalus borealis Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) Food Microbiology 30 1 173 179 |
institution |
Open Polar |
collection |
Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
op_collection_id |
ftccsdartic |
language |
English |
topic |
RAPQUANTITATIVE DETECTION STAA medium Seafood Spoilage Brochothrix thermosphacta Real-time PCR 16S rRNA COLD-SMOKED SALMON LISTERIA-MONOCYTOGENES PROPIDIUM MONOAZIDE PANDALUS-BOREALIS MICROBIAL ECOLOGY MEAT FOOD CARNOBACTERIUM LACTOCOCCUS [SDV.SA]Life Sciences [q-bio]/Agricultural sciences |
spellingShingle |
RAPQUANTITATIVE DETECTION STAA medium Seafood Spoilage Brochothrix thermosphacta Real-time PCR 16S rRNA COLD-SMOKED SALMON LISTERIA-MONOCYTOGENES PROPIDIUM MONOAZIDE PANDALUS-BOREALIS MICROBIAL ECOLOGY MEAT FOOD CARNOBACTERIUM LACTOCOCCUS [SDV.SA]Life Sciences [q-bio]/Agricultural sciences Mamlouk, Kelthoum, Mace, Sabrina, Guilbaud, Morgan, JAFFRES, Emmanuel, Ferchichi, Mounir, Prévost, Herve, Pilet, Marie-France, Dousset, Xavier Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR |
topic_facet |
RAPQUANTITATIVE DETECTION STAA medium Seafood Spoilage Brochothrix thermosphacta Real-time PCR 16S rRNA COLD-SMOKED SALMON LISTERIA-MONOCYTOGENES PROPIDIUM MONOAZIDE PANDALUS-BOREALIS MICROBIAL ECOLOGY MEAT FOOD CARNOBACTERIUM LACTOCOCCUS [SDV.SA]Life Sciences [q-bio]/Agricultural sciences |
description |
Brochothrix thermosphacta, a Cram-positive bacterium, is considered as the predominant spoilage microbiota of modified atmosphere packing (MAP) shrimp and fish. Traditional methods currently used to detect B. thermosphacta in foods are time-consuming and labour-intensive. The aim of this study was to develop a real-time PCR quantification method combined with a propidium monoazide (PMA) sample treatment step to monitor the population of B. thermosphacta in cooked shrimp and salmon. The specificity of the two primers M0405 and M0404 used to amplify a 70 bp fragment of the 16S rRNA gene was demonstrated by using purified DNA from 30 strains, among 21 bacterial species including 22 reference strains. Using these primers for real-time PCR and in pure culture, a good correlation was obtained between real-time PCR and the conventional plating method. Quantification was linear over 7-log units using artificially inoculated samples. The method performed successfully when tested on naturally contaminated cooked shrimp and fresh salmon, with a minimum threshold of 1.9 x 10(2) CFU/g for accurate quantification of B. the rmosphacta. The correlation between the B. thermosphacta counts obtained by real-time PCR and plate counts on naturally contaminated shrimp and salmon was high (R-2=0.895). Thus, this study presents a rapid tool for producing reliable quantitative data on B. thermosphacta in cooked shrimp and fresh salmon. (C) 2011 Elsevier Ltd. All rights reserved. |
author2 |
Oniris, Secalim UMR1014 LUNAM Université Nantes Angers Le Mans MICrobiologie de l'ALImentation au Service de la Santé (MICALIS) Institut National de la Recherche Agronomique (INRA)-AgroParisTech SECurité des ALIments et Microbiologie Institut National de la Recherche Agronomique (INRA) Region Pays de la Loire, France |
format |
Article in Journal/Newspaper |
author |
Mamlouk, Kelthoum, Mace, Sabrina, Guilbaud, Morgan, JAFFRES, Emmanuel, Ferchichi, Mounir, Prévost, Herve, Pilet, Marie-France, Dousset, Xavier |
author_facet |
Mamlouk, Kelthoum, Mace, Sabrina, Guilbaud, Morgan, JAFFRES, Emmanuel, Ferchichi, Mounir, Prévost, Herve, Pilet, Marie-France, Dousset, Xavier |
author_sort |
Mamlouk, Kelthoum, |
title |
Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR |
title_short |
Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR |
title_full |
Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR |
title_fullStr |
Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR |
title_full_unstemmed |
Quantification of viable Brochothrix thermosphacta in cooked shrimp and salmon by real-time PCR |
title_sort |
quantification of viable brochothrix thermosphacta in cooked shrimp and salmon by real-time pcr |
publisher |
HAL CCSD |
publishDate |
2012 |
url |
https://hal.archives-ouvertes.fr/hal-01004204 https://doi.org/10.1016/j.fm.2011.09.012 |
genre |
Pandalus borealis |
genre_facet |
Pandalus borealis |
op_source |
ISSN: 0740-0020 EISSN: 1095-9998 Food Microbiology https://hal.archives-ouvertes.fr/hal-01004204 Food Microbiology, Elsevier, 2012, 30 (1), pp.173 - 179. ⟨10.1016/j.fm.2011.09.012⟩ |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.fm.2011.09.012 hal-01004204 https://hal.archives-ouvertes.fr/hal-01004204 doi:10.1016/j.fm.2011.09.012 PRODINRA: 215388 WOS: 000300739900023 |
op_doi |
https://doi.org/10.1016/j.fm.2011.09.012 |
container_title |
Food Microbiology |
container_volume |
30 |
container_issue |
1 |
container_start_page |
173 |
op_container_end_page |
179 |
_version_ |
1766162323057147904 |