Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches

The aims of this study were i) to evaluate the possibility to detect and possibly quantify microorganisms belonging to different domains experimentally spiked in smoked salmon at known concentrations using shotgun metagenomics; ii) to compare the sequencing results using four bioinformatic tools. Th...

Full description

Bibliographic Details
Main Authors: Alessandra De Cesare, Chiara Oliveri, Alex Lucchi, Frederique Pasquali, Gerardo Manfreda
Format: Article
Language:English
Published: PAGEPress Publications 2019-12-01
Series:Italian Journal of Food Safety
Subjects:
Online Access:https://www.pagepressjournals.org/index.php/ijfs/article/view/8462
_version_ 1828486333978378240
author Alessandra De Cesare
Chiara Oliveri
Alex Lucchi
Frederique Pasquali
Gerardo Manfreda
author_facet Alessandra De Cesare
Chiara Oliveri
Alex Lucchi
Frederique Pasquali
Gerardo Manfreda
author_sort Alessandra De Cesare
collection DOAJ
description The aims of this study were i) to evaluate the possibility to detect and possibly quantify microorganisms belonging to different domains experimentally spiked in smoked salmon at known concentrations using shotgun metagenomics; ii) to compare the sequencing results using four bioinformatic tools. The salmon was spiked with six species of bacteria, including potential foodborne pathogens, as well as Cryptosporidium parvum, Saccharomyces cerevisiae and Bovine alphaherpesvirus 1. After spiking, the salmon was kept refrigerated before DNA extraction, library preparation and sequencing at 7 Gbp in paired ends at 150 bp. The bioinformatic tools named MG-RAST, OneCodex, CosmosID and MgMapper were used for the sequence analysis and the data provided were compared using STAMP. All bacteria spiked in the salmon were identified using all bioinformatic tools. Such tools were also able to assign the higher abundances to the species Propionibacterium freudenreichii spiked at the highest concentration in comparison to the other bacteria. Nevertheless, different abundances were quantified for bacteria spiked in the salmon at the same cell concentration. Cryptosporidium parvum was detected by all bioinformatics tools, while Saccharomyces cerevisiae by MG-RAST only. Finally, the DNA virus was detected by CosmosID and OneCodex only. Overall, the results of this study showed that shotgun metagenomics can be applied to detect microorganisms belonging to different domains in the same food sample. Nevertheless, a direct correlation between cell concentration of each spiked microorganism and number of corresponding reads cannot be established yet.
first_indexed 2024-12-11T09:31:00Z
format Article
id doaj.art-aeeda1bc6ee549d1aa8dc2803fb2c674
institution Directory Open Access Journal
issn 2239-7132
language English
last_indexed 2024-12-11T09:31:00Z
publishDate 2019-12-01
publisher PAGEPress Publications
record_format Article
series Italian Journal of Food Safety
spelling doaj.art-aeeda1bc6ee549d1aa8dc2803fb2c6742022-12-22T01:13:01ZengPAGEPress PublicationsItalian Journal of Food Safety2239-71322019-12-018410.4081/ijfs.2019.8462Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approachesAlessandra De Cesare0Chiara Oliveri1Alex Lucchi2Frederique Pasquali3Gerardo Manfreda4Department of Veterinary Medical Sciences, Alma Mater Studiorum University of BolognaDepartment of Agricultural and Food Sciences, Alma Mater Studiorum University of BolognaDepartment of Agricultural and Food Sciences, Alma Mater Studiorum University of BolognaDepartment of Agricultural and Food Sciences, Alma Mater Studiorum University of BolognaDepartment of Agricultural and Food Sciences, Alma Mater Studiorum University of BolognaThe aims of this study were i) to evaluate the possibility to detect and possibly quantify microorganisms belonging to different domains experimentally spiked in smoked salmon at known concentrations using shotgun metagenomics; ii) to compare the sequencing results using four bioinformatic tools. The salmon was spiked with six species of bacteria, including potential foodborne pathogens, as well as Cryptosporidium parvum, Saccharomyces cerevisiae and Bovine alphaherpesvirus 1. After spiking, the salmon was kept refrigerated before DNA extraction, library preparation and sequencing at 7 Gbp in paired ends at 150 bp. The bioinformatic tools named MG-RAST, OneCodex, CosmosID and MgMapper were used for the sequence analysis and the data provided were compared using STAMP. All bacteria spiked in the salmon were identified using all bioinformatic tools. Such tools were also able to assign the higher abundances to the species Propionibacterium freudenreichii spiked at the highest concentration in comparison to the other bacteria. Nevertheless, different abundances were quantified for bacteria spiked in the salmon at the same cell concentration. Cryptosporidium parvum was detected by all bioinformatics tools, while Saccharomyces cerevisiae by MG-RAST only. Finally, the DNA virus was detected by CosmosID and OneCodex only. Overall, the results of this study showed that shotgun metagenomics can be applied to detect microorganisms belonging to different domains in the same food sample. Nevertheless, a direct correlation between cell concentration of each spiked microorganism and number of corresponding reads cannot be established yet.https://www.pagepressjournals.org/index.php/ijfs/article/view/8462Shotgun metagenomics, smoked salmon, microbiological hazards, bioinformatic tools
spellingShingle Alessandra De Cesare
Chiara Oliveri
Alex Lucchi
Frederique Pasquali
Gerardo Manfreda
Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches
Italian Journal of Food Safety
Shotgun metagenomics, smoked salmon, microbiological hazards, bioinformatic tools
title Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches
title_full Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches
title_fullStr Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches
title_full_unstemmed Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches
title_short Application of shotgun metagenomics to smoked salmon experimentally spiked: Comparison between sequencing and microbiological data using different bioinformatic approaches
title_sort application of shotgun metagenomics to smoked salmon experimentally spiked comparison between sequencing and microbiological data using different bioinformatic approaches
topic Shotgun metagenomics, smoked salmon, microbiological hazards, bioinformatic tools
url https://www.pagepressjournals.org/index.php/ijfs/article/view/8462
work_keys_str_mv AT alessandradecesare applicationofshotgunmetagenomicstosmokedsalmonexperimentallyspikedcomparisonbetweensequencingandmicrobiologicaldatausingdifferentbioinformaticapproaches
AT chiaraoliveri applicationofshotgunmetagenomicstosmokedsalmonexperimentallyspikedcomparisonbetweensequencingandmicrobiologicaldatausingdifferentbioinformaticapproaches
AT alexlucchi applicationofshotgunmetagenomicstosmokedsalmonexperimentallyspikedcomparisonbetweensequencingandmicrobiologicaldatausingdifferentbioinformaticapproaches
AT frederiquepasquali applicationofshotgunmetagenomicstosmokedsalmonexperimentallyspikedcomparisonbetweensequencingandmicrobiologicaldatausingdifferentbioinformaticapproaches
AT gerardomanfreda applicationofshotgunmetagenomicstosmokedsalmonexperimentallyspikedcomparisonbetweensequencingandmicrobiologicaldatausingdifferentbioinformaticapproaches