Lactic acid bacteria in cow raw milk for cheese production: Which and how many?

Bettera, Luca; Levante, Alessia; Bancalari, Elena; Bottari, Benedetta; Gatti, Monica

doi:10.3389/fmicb.2022.1092224

Cited by 22 publications

(14 citation statements)

References 142 publications

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“…A common practice for cheese production when using pasteurized milk is the addition of commercial starter cultures to standardize the fermentation process and/or improve the sensory characteristics. However, these cultures are responsible for a flat organoleptic pattern rather than a rich traditional taste ( Leroy and De Vuyst, 2004 ; Psoni et al, 2006 ), so the addition of selected autochthonous LAB strains to cheese milk as adjunct cultures is gaining more attention, as it is reflected in the recent scientific literature ( de Souza and Dias, 2017 ; Gaglio et al, 2021 ; Bettera et al, 2022 ). These added microorganisms can release intracellular enzymes during ripening and storage, which mainly impact the nutritional, technological and sensory properties of cheeses ( Escobar-Zepeda et al, 2016 ; Fox et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Application of multi-functional lactic acid bacteria strains in a pilot scale feta cheese production

Kamarinou,

Papadopoulou,

Doulgeraki

et al. 2023

Front. Microbiol.

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Feta cheese is the most recognized Greek Protected Designation of Origin (PDO) product in the world. The addition of selected autochthonous lactic acid bacteria (LAB) strains to cheese milk as adjunct cultures is gaining more attention, since they can impact the nutritional, technological and sensory properties of cheeses, as well as improve the safety of the product. The aim of this study was to produce Feta cheese with enhanced quality and safety, and distinctive organoleptic characteristics by applying autochthonous lactic acid bacteria (LAB) with multi-functional properties as adjunct cultures. Feta cheeses were produced with the commercial lactococcal starter culture and the addition of 9 LAB strains (Lactococcus lactis SMX2 and SMX16, Levilactobacillus brevis SRX20, Lacticaseibacillus paracasei SRX10, Lactiplantibacillus plantarum FRX20 and FB1, Leuconostoc mesenteroides FMX3, FMX11, and FRX4, isolated from artisanal Greek cheeses) in different combinations to produce 13 cheese trials (12 Feta trials with the adjunct LAB isolates and the control trial). In addition, Feta cheese manufactured with FMX3 and SMX2 and control Feta cheese were artificially inoculated (4 log CFU/g) with Listeria monocytogenes (a cocktail of 4 acid or non-acid adapted strains). Cheese samples were monitored by microbiological and physicochemical analyses during ripening, and microbiological, physicochemical, molecular and sensory analyses during storage at 4°C. The results showed that after manufacture, the LAB population was ca. 9.0 log CFU/g at all samples, whereas during storage, their population declined to 6.5–7.0 log CFU/g. In the Listeria inoculated samples, Listeria was absent after 60 days (end of ripening) and after 90 days in the adjunct culture, and in the control trials, respectively. Moreover, the addition of selected strains, especially Lcb. paracasei SRX10, led to cheeses with desirable and distinctive organoleptic characteristics. Furthermore, randomly amplified polymorphic PCR (RAPD-PCR) molecular analysis confirmed that the multi-functional LAB strains were viable by the end of storage. Overall, the results of this study are promising for the use of autochthonous strains in various combinations with the commercial starter culture to satisfy industry requirements and consumer demands for traditional and high added value fermented products.

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Section: Introductionmentioning

confidence: 99%

Application of multi-functional lactic acid bacteria strains in a pilot scale feta cheese production

Kamarinou,

Papadopoulou,

Doulgeraki

et al. 2023

Front. Microbiol.

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“…While starter cultures are produced both at the laboratory or industrial (mixed defined-strain cultures), and artisanal (natural undefined strain culture) level, adjunct cultures with NSLAB are only commercialized as mixed defined-strain cultures. NSLAB are naturally present in raw milk, however, although some species may have very low abundance, since LAB are just a subdominant part of the raw milk microbiota ( Bettera et al, 2023 ). This makes their selection and isolation difficult.…”

Section: Discussionmentioning

confidence: 99%

Selective enrichment of the raw milk microbiota in cheese production: Concept of a natural adjunct milk culture

et al. 2023

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In cheese production, microorganisms are usually added at the beginning of the process as primary starters to drive curd acidification, while secondary microorganisms, with other pro-technological features important for cheese ripening, are added as selected cultures. This research aimed to investigate the possibilities of influencing and selecting the raw milk microbiota using artisanal traditional methods, providing a simple method to produce a natural supplementary culture. We investigated the production of an enriched raw milk whey culture (eRWC), a natural adjunct microbial culture produced from mixing an enriched raw milk (eRM) with a natural whey culture (NWC). The raw milk was enriched by spontaneous fermentation for 21 d at 10°C. Three milk enrichment protocols were tested: heat treatment before incubation, heat treatment plus salt addition, and no treatment. The eRMs were then co-fermented with NWC (ratio of 1:10) at 38°C for 6 h (young eRWC) and 22 h (old eRWC). Microbial diversity during cultures’ preparation was evaluated through the determination of colony forming units on selective growth media, and next-generation sequencing (16S rRNA gene amplicon sequencing). The enrichment step increased the streptococci and lactobacilli but reduced microbial richness and diversity of the eRMs. Although the lactic acid bacteria viable count was not significantly different between the eRWCs, they harbored higher microbial richness and diversity than NWC. Natural adjunct cultures were then tested in cheese making trials, following the microbial development, and assessing the chemical quality of the 120 d ripened cheeses. The use of eRWCs slowed the curd acidification in the first hours of cheese making but the pH 24 h after production settled to equal values for all the cheeses. Although the use of diverse eRWCs contributed to having a richer and more diverse microbiota in the early stages of cheese making, their effect decreased over time during ripening, showing an inferior effect to the raw milk microbiota. Even if more research is needed, the optimization of such a tool could be an alternative to the practice of isolating, geno-pheno-typing, and formulating mixed-defined-strain adjunct cultures that require knowledge and facilities not always available for artisanal cheese makers.

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“…casei group (LCG) and are supposed to arise from raw cow milk and dairy environment ( Bottari et al, 2018 ). LCG becomes dominant starting from 2 months of ripening ( Bottari et al, 2018 ; Bettera et al, 2023 ), due to their tolerance toward pH values as low as 4.9, salt concentrations up to 6%, and a wide range of temperatures (2–53°C) ( Settanni and Moschetti, 2010 ; Gobbetti et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Cultivable microbial diversity, peptide profiles, and bio-functional properties in Parmigiano Reggiano cheese

Martini,

Sola,

Cattivelli

et al. 2024

Front. Microbiol.

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IntroductionLactic acid bacteria (LAB) communities shape the sensorial and functional properties of artisanal hard-cooked and long-ripened cheeses made with raw bovine milk like Parmigiano Reggiano (PR) cheese. While patterns of microbial evolution have been well studied in PR cheese, there is a lack of information about how this microbial diversity affects the metabolic and functional properties of PR cheese.MethodsTo fill this information gap, we characterized the cultivable fraction of natural whey starter (NWS) and PR cheeses at different ripening times, both at the species and strain level, and investigated the possible correlation between microbial composition and the evolution of peptide profiles over cheese ripening.Results and discussionThe results showed that NWS was a complex community of several biotypes belonging to a few species, namely, Streptococcus thermophilus, Lactobacillus helveticus, and Lactobacillus delbrueckii subsp. lactis. A new species-specific PCR assay was successful in discriminating the cheese-associated species Lacticaseibacillus casei, Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, and Lacticaseibacillus zeae. Based on the resolved patterns of species and biotype distribution, Lcb. paracasei and Lcb. zeae were most frequently isolated after 24 and 30 months of ripening, while the number of biotypes was inversely related to the ripening time. Peptidomics analysis revealed more than 520 peptides in cheese samples. To the best of our knowledge, this is the most comprehensive survey of peptides in PR cheese. Most of them were from β-caseins, which represent the best substrate for LAB cell-envelope proteases. The abundance of peptides from β-casein 38–88 region continuously increased during ripening. Remarkably, this region contains precursors for the anti-hypertensive lactotripeptides VPP and IPP, as well as for β-casomorphins. We found that the ripening time strongly affects bioactive peptide profiles and that the occurrence of Lcb. zeae species is positively linked to the incidence of eight anti-hypertensive peptides. This result highlighted how the presence of specific LAB species is likely a pivotal factor in determining PR functional properties.

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Lactic acid bacteria in cow raw milk for cheese production: Which and how many?

Cited by 22 publications

References 142 publications

Application of multi-functional lactic acid bacteria strains in a pilot scale feta cheese production

Application of multi-functional lactic acid bacteria strains in a pilot scale feta cheese production

Selective enrichment of the raw milk microbiota in cheese production: Concept of a natural adjunct milk culture

Cultivable microbial diversity, peptide profiles, and bio-functional properties in Parmigiano Reggiano cheese

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