Development and Application of Starter Cultures

A special issue of Fermentation (ISSN 2311-5637). This special issue belongs to the section "Fermentation for Food and Beverages".

Deadline for manuscript submissions: closed (31 December 2023) | Viewed by 15025

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Agris Sardegna, Agenzia Regionale per la Ricerca in Agricoltura, Associated Member of the JRU MIRRI-IT, Loc. Bonassai, SS291 km 18.600, 07100 Sassari, Italy
Interests: food microbiology; fermented food; microbial biodiversity; microbial culture collections; lactic acid bacteria; starter cultures; fingerprint; antibiotic resistance
Special Issues, Collections and Topics in MDPI journals

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Guest Editor
Agris Sardegna, Agenzia Regionale per la Ricerca in Agricoltura, Associated Member of the JRU MIRRI-IT, Loc. Bonassai, SS291 km 18.600, 07100 Sassari, Italy
Interests: food microbiology; fermented food; microbial biodiversity; microbial culture collections; lactic acid bacteria; starter cultures; fingerprint; antibiotic resistance
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

In recent decades, starter cultures have been developed to aid raw material processing with the aim of obtaining different types of fermented food products. In fact, properly developed selected starters represent a convenient solution to easily and safely carry out the fermentation when the concentration of the microbiota colonizing the production environment and the raw material itself turns out to be inadequate, or natural starter cultures are difficult to obtain and manage.

Different purposes can be pursued when developing a starter culture: a) selecting a low, defined number of species/strains, on the basis of their strong aptitude to fulfil the biochemical processes required by each production technology and for their suitability to be grown in the laboratory, or b) trying to reproduce autochthonous biodiverse natural cultures where an indefinite number of species and strains, starter and nonstarter (but crucial during the whole fermentation and ripening of food), coexist in equilibrium. Both choices have pros and cons.

On one hand, selected cultures, because of and despite their high technological efficiency, easily become the dominant microbiota of the product, causing a dramatic decrease in microbial biodiversity and loss of peculiar sensory characteristics of fermented food. In fact, this kind of cultures are widely applied, at high concentration, to industrial level productions that do not possess geographic niches and typicity.

On the contrary, natural microbial communities have a strains composition which is not reproducible in any place other than that of their origin, contributing to preserve microbial biodiversity and enriching products with peculiar sensory features that bind them to the territory. Indeed, autochthonous natural starter cultures usually characterize the most typical and high quality agri-food products. However, their technological performance is not standardized and their use is not risk-free since, together with useful autochthonous microorganisms, even pathogen or spoilage ones could be potentially inoculated and allowed to contaminate the product.

The goal of this Special Issue is to host innovative or review papers facing the challenge of developing starter cultures or, at least, laying the foundation for them, which could be applicable at artisanal, pilot or industrial scale, able to guaranteeing safety, quality constancy, technological performances reproducibility, preserving biodiversity, and peculiar sensory characteristics usually linked to traditional products, while overcoming the problems associated with the daily propagation of natural cultures. Furthermore, papers dealing with if and how fermented products consumption could affect human gastro-intestinal tract microbiota, and eventually the health of the consumers, would be welcome.

Dr. Roberta Comunian
Dr. Luigi Chessa
Guest Editors

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Keywords

  • starter cultures
  • fermented food
  • microbial biodiversity
  • metagenomics
  • traditional foods
  • food safety
  • antibiotic resistance
  • microbial culture collections
  • gut microbiota
  • probiotics

Published Papers (10 papers)

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Research

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15 pages, 946 KiB  
Article
Biodiversity and Safety: Cohabitation Experimentation in Undefined Starter Cultures for Traditional Dairy Products
by Luigi Chessa, Elisabetta Daga, Ilaria Dupré, Antonio Paba, Maria C. Fozzi, Davide G. Dedola and Roberta Comunian
Fermentation 2024, 10(1), 29; https://doi.org/10.3390/fermentation10010029 - 29 Dec 2023
Viewed by 1287
Abstract
Natural starter cultures, characterised by undefined microbiota, can contribute to the technological process, giving peculiar characteristics to artisanal fermented foods. Several species have a long history of safe use and have obtained Qualified Presumption of Safety (QPS) status from the European Food Safety [...] Read more.
Natural starter cultures, characterised by undefined microbiota, can contribute to the technological process, giving peculiar characteristics to artisanal fermented foods. Several species have a long history of safe use and have obtained Qualified Presumption of Safety (QPS) status from the European Food Safety Authority (EFSA), whereas others (non-QPS) could represent a potential risk for consumers’ health and must undergo a safety assessment. In this work, the biodiversity, at species and strain level, by pulsed-field gel electrophoresis (PFGE) and (GTG)5 rep-PCR, of an undefined natural starter culture, in frozen and lyophilized form, obtained from ewe’s raw milk avoiding thermal treatment or microbial selection, was investigated. The culture was constituted by different biotypes of Enterococcus durans, Enterococcus faecium, Enterococcus faecalis, and Lacticaseibacillus paracasei. Streptococcus oralis and Streptococcus salivarius were also found, over species belonging to the Streptococcus bovisStreptococcus equinus complex (SBSEC), like Streptococcus gallolyticus subsp. macedonicus, Streptococcus lutetiensis, and Streptococcus equinus. Molecular investigation on virulence and antibiotic resistance genes, as well as minimum inhibitory concentration (MIC) determination, revealed that all the non-QPS strains can be considered safe in the perspective of using this culture for cheesemaking. The obtainment of a natural culture directly from ewe’s raw milk bypassing thermal treatment and selection of pro-technological bacteria can be advantageous in terms of biodiversity preservation, but non-QPS microorganisms can be included in the natural starter and also in cheeses, especially in traditional ones obtained from fermenting raw milk. Following EFSA guidelines, artisanal factories should not be allowed to produce starter cultures by themselves from raw milk, running the risk of including some non-QPS species in their culture, and only selected starters could be used for cheesemaking. A revision of the criteria of QPS guidelines should be necessary. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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18 pages, 1633 KiB  
Article
Screening of Acetic Acid Bacteria Isolated from Various Sources for Use in Kombucha Production
by Dong-Hun Lee, Su-Hwan Kim, Chae-Yun Lee, Hyeong-Woo Jo, Won-Hee Lee, Eun-Hye Kim, Byung-Kuk Choi and Chang-Ki Huh
Fermentation 2024, 10(1), 18; https://doi.org/10.3390/fermentation10010018 - 26 Dec 2023
Cited by 1 | Viewed by 1791
Abstract
The objective of this study was to isolate and identify strains of Acetobacter suitable for use in the development of a complex microbial culture for producing Kombucha and to examine the fermentation characteristics for selection of suitable strains. A medium supplemented with calcium [...] Read more.
The objective of this study was to isolate and identify strains of Acetobacter suitable for use in the development of a complex microbial culture for producing Kombucha and to examine the fermentation characteristics for selection of suitable strains. A medium supplemented with calcium carbonate was used for isolation of acetic acid bacteria from 22 various sources. Colonies observed in the clear zone resulting from decomposition of calcium carbonate by acid produced by microorganisms were collected. Identification of the collected strains was based on biological and morphological characteristics, and the results of base sequence analysis. A total of 37 strains were identified, including six species in the Acetobacter genus: Acetobacter pasteurianus, Acetobacter orientalis, Acetobacter cibinongensis, Acetobacter pomorum, Acetobacter ascendens, and Acetobacter malorum, as well as one species in the Gluconobacter genus, Gluconobacter oxydans. Among thirty-seven strains, seven strains of acetic acid bacteria with exceptional acid and alcohol tolerance were selected, and an evaluation of their fermentation characteristics according to fermentation temperature and period was performed. The results showed a titratable acidity of 1.68% for the Acetobacter pasteurianus SFT-18 strain, and an acetic acid bacteria count of 9.52 log CFU/mL at a fermentation temperature of 35 °C. The glucuronic acid and gluconate contents for the Gluconobacter oxydans SFT-27 strain were 10.32 mg/mL and 25.49 mg/mL, respectively. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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27 pages, 4872 KiB  
Article
Functional Characterization of Saccharomyces Yeasts from Cider Produced in Hardanger
by Urban Česnik, Mitja Martelanc, Ingunn Øvsthus, Tatjana Radovanović Vukajlović, Ahmad Hosseini, Branka Mozetič Vodopivec and Lorena Butinar
Fermentation 2023, 9(9), 824; https://doi.org/10.3390/fermentation9090824 - 08 Sep 2023
Cited by 2 | Viewed by 986
Abstract
Saccharomyces cerevisiae is commonly used for the production of alcoholic beverages, including cider. In this study, we examined indigenous S. cerevisiae and S. uvarum strains, both species commonly found in cider from Hardanger (Norway), for their strain-specific abilities to produce volatile and non-volatile [...] Read more.
Saccharomyces cerevisiae is commonly used for the production of alcoholic beverages, including cider. In this study, we examined indigenous S. cerevisiae and S. uvarum strains, both species commonly found in cider from Hardanger (Norway), for their strain-specific abilities to produce volatile and non-volatile compounds. Small-scale fermentation of apple juice with 20 Saccharomyces strains was performed to evaluate their aroma-producing potential as a function of amino acids (AAs) and other physicochemical parameters under the same experimental conditions. After fermentation, sugars, organic acids, AAs, and biogenic amines (BAs) were quantified using the HPLC–UV/RI system. A new analytical method was developed for the simultaneous determination of nineteen AAs and four BAs in a single run using HPLC–UV with prior sample derivatization. Volatile compounds were determined using HS-SPME-GC-MS. Based on 54 parameters and after the removal of outliers, the nineteen strains were classified into four groups. In addition, we used PLS regression to establish a relationship between aroma compounds and predictor variables (AAs, BAs, organic acids, sugars, hydrogen sulfide (H2S) production, CO2 release) of all 19 strains tested. The results of the VIP show that the main predictor variables affecting the aroma compounds produced by the selected yeasts are 16, belonging mainly to AAs. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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14 pages, 3953 KiB  
Article
Effects of Main Nutrient Sources on Improving Monascus Pigments and Saccharifying Power of Monascus purpureus in Submerged Fermentation
by Yingying Huang, Jiashi Chen, Qing Chen and Chenglong Yang
Fermentation 2023, 9(7), 696; https://doi.org/10.3390/fermentation9070696 - 24 Jul 2023
Viewed by 792
Abstract
Hong Qu (HQ), obtained through fermentation of various grains using Monascus spp., has been widely utilized as the main and characteristic initial saccharification and traditional fermentation starter in the food brewing industry. The quality, color, and flavor of HQ and HQ wine are [...] Read more.
Hong Qu (HQ), obtained through fermentation of various grains using Monascus spp., has been widely utilized as the main and characteristic initial saccharification and traditional fermentation starter in the food brewing industry. The quality, color, and flavor of HQ and HQ wine are closely related to the saccharifying power (SP) and Monascus pigments (MPs) of Monascus spp. In this study, to optimize the culture medium in submerged fermentation by M. purpureus G11 for improving SP and MPs, the effects of carbon source, nitrogen source, inorganic salts, and vitamins on SP activity and biosynthesis of MPs were explored through single-factor analysis and response surface Box–Behnken experiments. The results showed that the optimal medium composition was 6.008% rice powder, 1.021% peptone, 0.0049% CuSO4, and 0.052% vitamin B1. Validation experiments performed under the optimized fermentation conditions showed a significant increase in MPs and SP by 14.91% and 36.24%, with maximum MPs and SP reaching 112.61 and 365.12 u/mL, respectively. This study provides a theoretical basis for enhancing MPs and SP in M. purpureus for HQ production, to improve the production efficiency and shorten the production cycle of HQ-related fermentation products. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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14 pages, 2002 KiB  
Article
Autochthonous Microbes to Produce Ligurian Taggiasca Olives (Imperia, Liguria, NW Italy) in Brine
by Grazia Cecchi, Simone Di Piazza, Ester Rosa, Furio De Vecchis, Milena Sara Silvagno, Junio Valerio Rombi, Micaela Tiso and Mirca Zotti
Fermentation 2023, 9(7), 680; https://doi.org/10.3390/fermentation9070680 - 19 Jul 2023
Viewed by 816
Abstract
Table olives are considered high-quality food, and Italy has a wealth of varieties and typical features that are truly unique in the world (about eighty cultivars of table olives or dual-purpose olives, four of which are protected by the protected designation of origin—PDO), [...] Read more.
Table olives are considered high-quality food, and Italy has a wealth of varieties and typical features that are truly unique in the world (about eighty cultivars of table olives or dual-purpose olives, four of which are protected by the protected designation of origin—PDO), and it is the second largest European consumer, behind Spain. The Taggiasca olive does not have a PDO, but it is very appreciated not only in the region of production (Liguria), but also in all the Italian regions and abroad. Autochthonous microbes (bacteria, yeasts, and filamentous fungi) are essential in the fermentative processes for brine olive production. However, these microbial communities that colonised the olive drupes are affected by the environmental conditions and the fermentation treatments. Hence the importance of studying and comparing olive microbes from different farms and investigating the relationships between bacteria, yeasts, and filamentous fungi to speed up the deamarisation process. Our results showed that yeasts are dominant relative to lactobacteria in all three brines studied, and Wickerhamomyces anomalus was the most performant fungus for the oleuropein degradation. The latter represents the best candidate for the realisation of a microbial starter. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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20 pages, 4682 KiB  
Article
Exploring the Inhibitory Activity of Selected Lactic Acid Bacteria against Bread Rope Spoilage Agents
by Giovanna Iosca, Joanna Ivy Irorita Fugaban, Süleyman Özmerih, Anders Peter Wätjen, Rolf Sommer Kaas, Quốc Hà, Radhakrishna Shetty, Andrea Pulvirenti, Luciana De Vero and Claus Heiner Bang-Berthelsen
Fermentation 2023, 9(3), 290; https://doi.org/10.3390/fermentation9030290 - 16 Mar 2023
Cited by 5 | Viewed by 1919
Abstract
In this study, a wide pool of lactic acid bacteria strains deposited in two recognized culture collections was tested against ropy bread spoilage bacteria, specifically belonging to Bacillus spp., Paenibacillus spp., and Lysinibacillus spp. High-throughput and ex vivo screening assays were performed to [...] Read more.
In this study, a wide pool of lactic acid bacteria strains deposited in two recognized culture collections was tested against ropy bread spoilage bacteria, specifically belonging to Bacillus spp., Paenibacillus spp., and Lysinibacillus spp. High-throughput and ex vivo screening assays were performed to select the best candidates. They were further investigated to detect the production of active antimicrobial metabolites and bacteriocins. Moreover, technological and safety features were assessed to value their suitability as biocontrol agents for the production of clean-label bakery products. The most prominent inhibitory activities were shown by four strains of Lactiplantibacillus plantarum (NFICC19, NFICC 72, NFICC163, and NFICC 293), two strains of Pediococcus pentosaceus (NFICC10 and NFICC341), and Leuconostoc citreum NFICC28. Moreover, the whole genome sequencing of the selected LAB strains and the in silico analysis showed that some of the strains contain operons for bacteriocins; however, no significant evidence was observed phenotypically. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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19 pages, 2436 KiB  
Article
Dynamics of Microbiota in Three Backslopped Liquid Sourdoughs That Were Triggered with the Same Starter Strains
by Valentina Tolu, Cristina Fraumene, Angela Carboni, Antonio Loddo, Manuela Sanna, Simonetta Fois, Tonina Roggio and Pasquale Catzeddu
Fermentation 2022, 8(10), 571; https://doi.org/10.3390/fermentation8100571 - 21 Oct 2022
Cited by 2 | Viewed by 1713
Abstract
The preparation of sourdough may include the use of starter microorganisms to address the fermentation process toward specific conditions. The aim of this work was to study the dynamics of the microbial ecosystem in three liquid sourdoughs (SD1, SD2 and SD3) triggered with [...] Read more.
The preparation of sourdough may include the use of starter microorganisms to address the fermentation process toward specific conditions. The aim of this work was to study the dynamics of the microbial ecosystem in three liquid sourdoughs (SD1, SD2 and SD3) triggered with the same microbial strains. Lactiplantibacillus plantarum (formerly known as Lactobacillus plantarum), Saccharomyces cerevisiae and Candida lambica strains were inoculated as starters, and sourdoughs were differentiated for the fermentation conditions and for the method of starter inoculation. The analyses were performed on the three sourdoughs propagated in the laboratory for 22 days and on the sample SD1, which was transferred to a bakery and refreshed over many months. The dynamics of microbial communities were studied by plate-count analysis and metataxonomic approach. The acidity of sourdough was evaluated over time. Metataxonomic analysis highlighted a large heterogeneity of fungi microbiota in all sourdough preparations, many of them probably originated from the flour, being pathogens of plants. Few yeast species were found, and S. cerevisiae was plentiful but did not predominate over the other species, whereas the C. lambica species decreased over time and then disappeared in all preparations. The bacterial microbiota was less heterogeneous than the fungi microbiota; the species L. plantarum, Leuconostoc citreum and Levilactobacillus brevis (formerly known as Lactobacillus brevis) were always present in all sourdoughs, whereas Fructilactobacillus sanfranciscensis (formerly known as Lactobacillus sanfranciscensis) became the dominant species in bakery-propagated SD1 and in SD2 at the end of the propagation period. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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Review

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14 pages, 875 KiB  
Review
The Microbial Community of Natural Whey Starter: Why Is It a Driver for the Production of the Most Famous Italian Long-Ripened Cheeses?
by Erasmo Neviani, Alessia Levante and Monica Gatti
Fermentation 2024, 10(4), 186; https://doi.org/10.3390/fermentation10040186 - 29 Mar 2024
Viewed by 480
Abstract
The remarkable global diversity in long-ripened cheese production can be attributed to the adaptability of the cheese microbiota. Most cheese types involve intricate microbial ecosystems, primarily represented by lactic acid bacteria (LAB). The present study aims to review the microbial community’s diversity in [...] Read more.
The remarkable global diversity in long-ripened cheese production can be attributed to the adaptability of the cheese microbiota. Most cheese types involve intricate microbial ecosystems, primarily represented by lactic acid bacteria (LAB). The present study aims to review the microbial community’s diversity in dairy fermentation processes, focusing on two famous Italian cheeses, Grana Padano and Parmigiano Reggiano, produced using natural whey starter (NWS). NWS, created by retaining whey from the previous day’s cheese batches, forms a microbiological connection between daily cheese productions. Through this technique, a dynamic microbiota colonizes the curd and influences cheese ripening. The back-slopping method in NWS preparation ensures the survival of diverse biotypes, providing a complex microbial community in which interactions among microorganisms are critical to ensuring its technological functionality. As highlighted in this review, the presence of microbial cells alone does not guarantee technological relevance. Critical microorganisms can grow and colonize the curd and cheese. This complexity enables NWS to adapt to artisanal production technologies while considering variations in raw milk microbiota, inhibitory compounds, and manufacturing conditions. This critical review aims to discuss NWS as a key factor in cheese making, considering microbial communities’ ability to evolve under different selective pressures and biotic and abiotic stresses. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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18 pages, 1167 KiB  
Review
Underexplored Potential of Lactic Acid Bacteria Associated with Artisanal Cheese Making in Brazil: Challenges and Opportunities
by Bianca de Oliveira Hosken, Gilberto Vinícius Melo Pereira, Thamylles Thuany Mayrink Lima, João Batista Ribeiro, Walter Coelho Pereira de Magalhães Júnior and José Guilherme Prado Martin
Fermentation 2023, 9(5), 409; https://doi.org/10.3390/fermentation9050409 - 25 Apr 2023
Cited by 2 | Viewed by 1608
Abstract
Artisanal cheeses are prepared using traditional methods with territorial, regional and cultural linkages. In Brazil, there is a great diversity of artisanal cheeses (BAC), which have historical, socioeconomic and cultural importance. The diversity of the BAC between producing regions is due to the [...] Read more.
Artisanal cheeses are prepared using traditional methods with territorial, regional and cultural linkages. In Brazil, there is a great diversity of artisanal cheeses (BAC), which have historical, socioeconomic and cultural importance. The diversity of the BAC between producing regions is due to the different compositions of raw milk, the steps involved in the process and the maturation time. The crucial step for cheese differentiation is the non-addition of starter cultures, i.e., spontaneous fermentation, which relies on the indigenous microbiota present in the raw material or from the environment. Therefore, each BAC-producing region has a characteristic endogenous microbiota, composed mainly of lactic acid bacteria (LAB). These bacteria are responsible for the technological, sensory and safety characteristics of the BAC. In this review, the biotechnological applications of the LAB isolated from different BAC were evidenced, including proteolytic, lipolytic, antimicrobial and probiotic activities. In addition, challenges and opportunities in this field are highlighted, because there are knowledge gaps related to artisanal cheese-producing regions, as well as the biotechnological potential. Thus, this review may provide new insights into the biotechnological applications of LAB and guide further research for the cheese-making process. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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18 pages, 772 KiB  
Review
Achievements of Autochthonous Wine Yeast Isolation and Selection in Romania—A Review
by Raluca-Ștefania Rădoi-Encea, Vasile Pădureanu, Camelia Filofteia Diguță, Marian Ion, Elena Brîndușe and Florentina Matei
Fermentation 2023, 9(5), 407; https://doi.org/10.3390/fermentation9050407 - 23 Apr 2023
Cited by 1 | Viewed by 1880
Abstract
Winemaking in Romania has a long-lasting history and traditions and its viticulture dates back centuries. The present work is focused on the development of wine yeast isolation and selection performed in different Romanian winemaking regions during past decades, presenting the advanement of the [...] Read more.
Winemaking in Romania has a long-lasting history and traditions and its viticulture dates back centuries. The present work is focused on the development of wine yeast isolation and selection performed in different Romanian winemaking regions during past decades, presenting the advanement of the methods and techniques employed, correlated with the impact on wine quality improvement. Apart from the historical side of such work, the findings will reveal how scientific advancement in the country was correlated with worldwide research in the topic and influenced local wines’ typicity. To create an overall picture of the local specificities, the work refers to local grape varieties and the characteristics of the obtained wines by the use of local yeasts as compared to commercial ones. Numerous autochthonous strains of Saccharomyces were isolated from Romanian vineyards, of which several demonstrated strong oenological characteristics. Meanwhile, different non-Saccharomyces yeast strains were also isolated and are nowadays receiving the attention of researchers seeking to develop new wines according to wine market tendencies and to support wine’s national identity. Full article
(This article belongs to the Special Issue Development and Application of Starter Cultures)
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