title: A method to isolate bacterial communities and characterize ecosystems from food products: Validation and utilization in as a reproducible chicken meat model.
abstract: Influenced by production and storage processes and by seasonal changes the diversity of meat products microbiota can be very variable. Because microbiotas influence meat quality and safety, characterizing and understanding their dynamics during processing and storage is important for proposing innovative and efficient storage conditions. Challenge tests are usually performed using meat from the same batch, inoculated at high levels with one or few strains. Such experiments do not reflect the true microbial situation, and the global ecosystem is not taken into account. Our purpose was to constitute live stocks of chicken meat microbiotas to create standard and reproducible ecosystems. We searched for the best method to collect contaminating bacterial communities from chicken cuts to store as frozen aliquots. We tested several methods to extract DNA of these stored communities for subsequent PCR amplification. We determined the best moment to collect bacteria in sufficient amounts during the product shelf life. Results showed that the rinsing method associated to the use of Mobio DNA extraction kit was the most reliable method to collect bacteria and obtain DNA for subsequent PCR amplification. Then, 23 different chicken meat microbiotas were collected using this procedure. Microbiota aliquots were stored at -80°C without important loss of viability. Their characterization by cultural methods confirmed the large variability (richness and abundance) of bacterial communities present on chicken cuts. Four of these bacterial communities were used to estimate their ability to regrow on meat matrices. Challenge tests performed on sterile matrices showed that these microbiotas were successfully inoculated and could overgrow the natural microbiota of chicken meat. They can therefore be used for performing reproducible challenge tests mimicking a true meat ecosystem and enabling the possibility to test the influence of various processing or storage conditions on complex meat matrices.
authors.affiliation: INRA, UMR1014, Secalim, Nantes, France; LUNAM Université, Oniris, Nantes, France.
authors.forename: Amélie
authors.initials: A
authors.lastname: Rouger
authors.affiliation: INRA, UMR1014, Secalim, Nantes, France; LUNAM Université, Oniris, Nantes, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Remenant
authors.affiliation: INRA, UMR1014, Secalim, Nantes, France; LUNAM Université, Oniris, Nantes, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Prévost
authors.affiliation: INRA, UMR1014, Secalim, Nantes, France; LUNAM Université, Oniris, Nantes, France. Electronic address: monique.zagorec@oniris-nantes.fr.
authors.forename: Monique
authors.initials: M
authors.lastname: Zagorec
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002645:Chickens / Q000382:microbiology*; D015169:Colony Count, Microbial; D005516:Food Microbiology / Q000379:methods*; D061353:Food Storage; D008460:Meat / Q000382:microbiology*; D064307:Microbiota
keywords: Challenge-test; DNA extraction; Microbiota; Poultry meat