CoChat
Sign Up
Cited by 45Probiotics and Fermented FoodsIdentification and Quantification in FoodAdvanced Scientific Research Methods

Development of a multiplex real time PCR to detect thermophilic lactic acid bacteria in natural whey starters.

Bottari, Benedetta, Agrimonti, Caterina, Gatti, Monica, Neviani, Erasmo, Marmiroli, Nelson·2013·International Journal of Food Microbiology
DOI 10.1016/j.ijfoodmicro.2012.10.011
Open this paper in CoChat — free account

Abstract

A multiplex real time PCR (mRealT-PCR) useful to rapidly screen microbial composition of thermophilic starter cultures for hard cooked cheeses and to compare samples with potentially different technological properties was developed. Novel primers directed toward pheS gene were designed and optimized for multiple detection of Lactobacillus helveticus, Lactobacillus delbrueckii, Streptococcus thermophilus and Lactobacillus fermentum. The assay was based on SYBR Green chemistry followed by melting curves analysis. The method was then evaluated for applications in the specific detection of the 4 lactic acid bacteria (LAB) in 29 different natural whey starters for Parmigiano Reggiano cheese production. The results obtained by mRealT-PCR were also compared with those obtained on the same samples by Fluorescence in Situ Hybridization (FISH) and Length-Heterogeneity PCR (LH-PCR). The mRealT-PCR developed in this study, was found to be effective for analyzing species present in the samples with an average sensitivity down to less than 600 copies of DNA and therefore sensitive enough to detect even minor LAB community members of thermophilic starter cultures. The assay was able to describe the microbial population of all the different natural whey starter samples analyzed, despite their natural variability. A higher number of whey starter samples with S. thermophilus and L. fermentum present in their microbial community were revealed, suggesting that these species could be more frequent in Parmigiano Reggiano natural whey starter samples than previously shown. The method was more effective than LH-PCR and FISH and, considering that these two techniques have to be used in combination to detect the less abundant species, the mRealT-PCR was also faster. Providing a single step sensitive detection of L. helveticus, L. delbrueckii, S. thermophilus and L. fermentum, the developed mRealT-PCR could be used for screening thermophilic starter cultures and to follow the presence of those species during ripening of derived dairy products. A major increase in understanding the starter culture contribution to cheese ecosystem could be harnessed to control cheese ripening and flavor formation.

Integrity signals

Registry

Crossref · 10.1016/j.ijfoodmicro.2012.10.011

Cites

36 of 36 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

This paper’s full text isn’t openly available, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • The presence of whole cells of L. helveticus and L. delbrueckii in curd samples confirmed what was previously observed, as these species come mainly from the natural whey starter and are frequently found at the beginning of ripening.

    “The presence of whole cells of L. helveticus and L. delbrueckii in curd samples confirmed what was previously observed, as these species come mainly from the natural whey starter and are frequently found at the beginning of ripening (Bottari et al., 2013; Gatti et al., 2014).”

    From The Interrelationship Between Microbiota and Peptides During Ripening as a Driver for Parmigiano Reggiano Cheese Quality (2020) · cites this paper as Bottari et al., 2013

  • The small differences among dairies at curd stage are likely due to the prevalence of SLAB coming from the starter, which generally contains mainly L. helveticus and L. delbrueckii and possibly S. thermophilus and L. fermentum.

    “This is likely due to the prevalence of SLAB coming from the starter, which, although prepared differently by each dairy, generally contains mainly L. helveticus and L. delbrueckii and possibly S. thermophilus and L. fermentum (Bottari et al., 2010, 2013; De Filippis et al., 2014; Bertani et al., 2020).”

    From The Interrelationship Between Microbiota and Peptides During Ripening as a Driver for Parmigiano Reggiano Cheese Quality (2020) · cites this paper as Bottari et al., 2013

  • The abundance of αS1-CN f(1-23) is positively correlated with the total amount of DNA from entire cells of L. helveticus and L. delbrueckii.

    “Interestingly, the abundance of αS1-CN f(1–23) – characteristic of the first stages of cheese manufacturing, within 48 h, thus derived from the proteolytic actions happening in the curd – is positively correlated with the total amount of DNA from entire cells of L. helveticus and L. delbrueckii, two species typical of PR natural whey starters (Bottari et al., 2010, 2013; De Filippis et al., 2014;”

    From The Interrelationship Between Microbiota and Peptides During Ripening as a Driver for Parmigiano Reggiano Cheese Quality (2020) · cites this paper as Bottari et al., 2013

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Bottari, B., Agrimonti, C., Gatti, M., Neviani, E., & Marmiroli, N.
Affiliations
University of Parma (IT)
Corresponding author
Gatti, Monica
ORCID
Venue
International Journal of Food Microbiology
Publisher
Elsevier BV
Published
2013-01
Volume
160
Issue
3
Pages
290-297
ISSN
0168-1605 (ISSN-L)
DOI
10.1016/j.ijfoodmicro.2012.10.011
OpenAlex
W2087597620
Cited by
45
References
36
Citations by year
2025: 6 · 2024: 5 · 2023: 1 · 2022: 1 · 2021: 4 · 2020: 5 · 2019: 4 · 2018: 2 · 2017: 6 · 2016: 4 · 2015: 1 · 2014: 1 · 2013: 4
FWCI
2.95
Type
journal article
Language
en
Update notices
erratum (2019-09-16)
Open access
closed
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • International Journal of Food Microbiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Teagasc
  • Regione Emilia-Romagna
Topics
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Identification and Quantification in Food
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Advanced Scientific Research Methods
Keywords
multiplex real-time PCR, thermophilic lactic acid bacteria, whey starter cultures, Parmigiano-Reggiano cheese, Lactobacillus helveticus, Lactobacillus delbrueckii, Streptococcus thermophilus, Lactobacillus fermentum, cheese ripening
MeSH terms
Cheese, Food Microbiology, Lactobacillus, Sensitivity and Specificity, Polymerase Chain Reaction, In Situ Hybridization, Fluorescence, Streptococcus thermophilus, Real-Time Polymerase Chain Reaction

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review