Spontaneous Fermentation Process Research Articles

Effect of spontaneous micro-fermentation on the physicochemical characteristics of Theobroma cacao L. beans

Objective: To evaluate the physicochemical characteristics of cacao beans during the spontaneous fermentation process in tepemixtle wooden boxes of three different sizes. Design/Methodology/Approach: Trinitarian-type cacao (Theobroma cacao L.) fruits were collected in a commercial orchard. The beans were fermented in boxes with the following operating conditions: volume of cacao with pulp, box size, a six-day period, and uncontrolled temperature and pH. The experiments were carried out in triplicate and an analysis of variance and a multiple comparison of means were performed with the Tukey test (p<0.05) using the R program. Results: The parameters evaluated had the same behavior in the 4 kg box than in the 25 kg box and the 60 kg box, which indicates that the physicochemical and quality characteristics of cacao beans are not impacted by the decrease in the box size. Therefore, smaller boxes are viable for small-scale fermentations. Study Limitations/Implications: Only cacao beans from the Trinitarian genetic group were evaluated. Findings/Conclusions: Small-scale fermentations can be an option for the evaluation of varieties when the beans must be fermented for the sensory analysis of their qualitative attributes. This micro-fermentation methodology can also be incorporated into genetic improvement programs or the evaluation of the processes aimed at their optimization.

Development of a defined mixed starter culture for the improvement of Tej, Ethiopian honey wine.

Ethiopian honey wine, Tej, is the most popular traditionally fermented alcoholic beverage in Ethiopia. Owing to the spontaneous fermentation process, the final product is neither predictable nor consistent in quality. Attempts have not been made before to solve this problem. Thus, the aim of this study was to develop a potential mixed starter culture of yeast and lactic acid bacteria isolated from Tej samples for the production of Tej with consistent quality. One hundred seventy-seven lactic acid bacteria and 194 yeasts were isolated from 30 Tej samples collected from southwest Ethiopia. After sequentially testing the isolates towards physiological stress tolerance and desired metabolic products, 10 lactic acid bacteria and 10 yeast isolates were screened. Later, four lactic acid bacteria and four yeast isolates were found to be compatible in co-culture tests. Finally, the combination of lactic acid bacteria and yeast isolates was formulated using the design of expert version 7.0.0 software, and six formulates (F #1-6) were designed. Controlled Tej fermentation was performed under laboratory conditions using six lactic acid bacteria-yeast starter culture formulations. The sensory attributes, in terms of color, flavor, odor, turbidity, and overall acceptance analysis scored 4.8/5.0 (F #2) and 4.7/5.0 (F #6), with mean significant variations (p < 0.05) among the other formulates. These two formulates were considered the best-mixed starter cultures compared to the control and other formulates. Matrix-assisted laser desorption ionization-time of flight analysis revealed that the lactic acid bacteria starters (AAUL7 and AAUL10) belonged to Lactobacillus paracasei. While the yeast starters (AAUY2 and AAUY8) belonged to Saccharomyces cerevisiae. These mixed lactic acid bacteria-yeast starter cultures could be used as the best starter culture for the fermentation of Ethiopian honey wine, Tej, with consistent quality.

Impact of immature coffee fruits and water addition during spontaneous fermentation process: Chemical composition and sensory profile

BackgroundCoffee fermentation process influences the final coffee composition and the sensory aspects which define the quality of the beverage. In this study, coffee fruits underwent spontaneous self-induced anaerobic fermentation using samples with two percentages of immature fruits in submerged and solid-state processing. The effects on the physicochemical composition and sensory quality of coffees were evaluated. ResultsThe two percentages of immature fruits corresponded to 11.0 and 0.3% of unripe fruits. The percentage of immature fruits significantly altered the initial content of sugars (sucrose, glucose, and fructose), ash, and titratable acidity. The water addition during the fermentative process did not significantly influence final moisture, proteins, citric acid, and propionic acid concentrations. Compared to the solid-state, the submerged process gave rise to coffees with lower concentrations of ethanol, glycerol, ash, lipids, succinic, and acetic acids. Coffee fermented with 0.3% of immature fruits showed higher lactic acid production in submerged fermentation (67.44 mg/g), and higher concentrations of ethanol (42.84 mg/g) and glycerol (1.68 mg/g) in solid-state fermentation. All coffees produced were classified as specialty coffees with a score above 84 points. However, the submerged fermented coffee with 11% immature fruit stood out with notes of caramel, brown sugar, honey, orange, lemon, floral, nut, yellow and red fruits. ConclusionsThis study confirmed that spontaneous fermentation can be used to produce specialty coffees. Differentiation in sensory attributes can be achieved through the addition of water and varying the percentage of green fruits during the fermentation process. Up to 11% of immature fruits did not compromise coffee quality.How to cite: Ferreira LJC, Casé IN, Bertarini PLL, et al. Impact of immature coffee fruits and water addition during spontaneous fermentation process: Chemical composition and sensory profile. Electron J Biotechnol 2024;69. https://doi.org/10.1016/j.ejbt.2024.04.001.

Combined omics expose microbial niches of fungi and bacteria correlating with wine volatile profiles in Douro wine region

Wine microbial communities establish complex ecological ecosystems that modulate the formation of aroma compounds, but only a few studies sought for correlations between specific microorganisms and wine volatiles. The present study combined metabarcoding and metabolomics for identifying microbial niches of fungi and bacteria correlating with the volatile profiles of wines of 3 renowned cultivars of the emblematic Douro region. Three major microbial niches were identified throughout the spontaneous fermentation processes, and the Hanseniaspora-Saccharomyces succession timing was cultivar-dependent. The largest niche included Hanseniaspora, Aureobasidium, Alternaria, Rhodotorula, Sporobolomyces, Massilia, Bacillus, Staphylococcus and Cutibacterium, that positively correlated with 7 metabolites, namely, acetoin, isoamyl acetate, ethyl propanoate, c-3-hexenol, phenylethyl acetate and 4-ethylphenol. The fermentative yeasts S. cerevisiae, Torulaspora delbrueckii and Meyerozyma caribbica strongly correlated with γ-butyrolactone, t-whiskylactone, isoamyl alcohol, ethyl decanoate, ethyl isobutyrate, diethyl succinate, isovaleric acid, 4-ethylguaiacol and 4-propylguaiacol. Lachancea quebecensis clustered with several pathogenic fungi (Penicillium citrinum, Erysiphe necator, Sclerotinia sclerotiorum, Aspergillus, Mycosphaerella tassiana) and bacteria (Pseudomonas spp., Bacteroides acidifaciens, Pantoea, Stenotrophomonas and Enhydrobacter), correlating positively with various monoterpenols and norisoprenoids including linalool and β-ionone, besides with benzyl alcohol, diacetyl, isobutyl acetate, ethyl-vanillate and methyl vanillinate. Metabolite-microbiota correlations denoted cultivar specificities likely underlying regional aromatic signatures.

Fungal Community Composition and Its Relationship with Volatile Compounds during Spontaneous Fermentation of Cabernet Sauvignon from Two Chinese Wine-Growing Regions.

The microbial community structure associated with wine in a wine-growing region is shaped by diverse ecological factors within that region, profoundly impacting the wine flavor. In wine fermentation, fungi contribute more sensory-active biochemical compounds than bacteria. In this study, we employed amplicon sequencing to measure samples from the spontaneous fermentation process of cabernet sauvignon wines from two wine-growing regions in China to study the diversity and structural evolution of fungi during spontaneous fermentation and analyze the correlation between fungi and volatile compounds. The results showed significant differences in fungal community structure and diversity in cabernet sauvignon musts from different geographical origins, and these differences affected the flavor quality of the wines. As alcoholic fermentation progressed, Saccharomyces became the dominant fungal genus and reshaped the fungal community structure, and the diversity of the fungal community decreased. However, the fungal communities of each wine-growing region remained distinct throughout the fermentation process. Furthermore, the correlation between the fungal community and volatile compounds indicated that wine is a product of fermentation involving multiple fungal genera, and the flavor is influenced by a variety of fungi. Our study enhances the comprehension of fungal communities in Chinese wine-growing regions, explaining the regulatory role of wine-related fungal microorganisms in wine flavor.

Integrating shotgun metagenomics and metabolomics to elucidate the dynamics of microbial communities and metabolites in fine flavor cocoa fermentation in Hainan

The aim of this study was to investigate the dynamic profile of microorganisms and metabolites in Hainan Trinitario cocoa during a six-day spontaneous box fermentation process. Shotgun metagenomic and metabolomic approaches were employed for this investigation. The potential metabolic functions of microorganisms in cocoa fermentation were revealed through a joint analysis of microbes, functional genes, and metabolites. During the anaerobic fermentation phase, Hanseniaspora emerged as the most prevalent yeast genus, implicated in pectin decomposition and potentially involved in glycolysis and starch and sucrose metabolism. Tatumella, possessing potential for pyruvate kinase, and Fructobacillus with a preference for fructose, constituted the primary bacteria during the pre-turning fermentation stage. Upon the introduction of oxygen into the fermentation mass, acetic acid bacteria ascended to dominant within the microflora. The exponential proliferation of Acetobacter resulted in a decline in taxonomic richness and abundance. Moreover, the identification of novel species within the Komagataeibacter genus suggests that Hainan cocoa may serve as a valuable reservoir for the discovery of unique cocoa fermentation bacteria. The KEGG annotation of metabolites and enzymes also highlighted the significant involvement of phenylalanine metabolism in cocoa fermentation. This research will offer a new perspective for the selection of starter strains and the formulation of mixed starter cultures.

Identification, semi-quantification, and dynamics of volatile organic compounds during spontaneous fermentation of two cocoa varieties from northern Peru, using HS-SPME/GC-MS techniques

During spontaneous fermentation, an important process through which cocoa beans pass, volatile organic compounds (VOCs) and non-VOCs are generated and act as precursors of aromas and flavors that influence the chocolate. In the north of Peru, cocoas of different varieties and qualities are grown, and one of the most valued is the fine aroma native cocoa (Cacao Fino de Aroma (CFA)) compared with the Castro Naranjal 51 Collection (CCN-51) variety. In this investigation, the VOCs generated during the spontaneous fermentation of CFA and CCN-51 varieties, from the department of Amazonas, Peru, were analyzed. For the extraction of VOCs, the headspace (HS) and solid-phase microextraction (SPME) techniques were used, while gas chromatography coupled to a mass spectrophotometer (GC-MS) was employed for their subsequent separation and identification. The spontaneous fermentation process lasted for 156 h, and a total of 122 VOCs were identified in 2 varieties. The dynamics of 20 VOCs were assessed, and the principal component analysis of 22 VOCs was performed to compare the profiles of VOC of the 2 cocoa varieties. In conclusion, the CFA variety presented higher concentrations of floral, fruity, and sweet aromatic VOCs that positively influence the quality of the chocolate.

Physical–Chemical and Metataxonomic Characterization of the Microbial Communities Present during the Fermentation of Three Varieties of Coffee from Colombia and Their Sensory Qualities

The microbial composition and physical-chemical characteristics were studied during the coffee fermentation of three Coffea arabica L. varieties, Var. Tabi, Var. Castillo General® and Var. Colombia. Mucilage and washed coffee seeds samples were collected at different stages of fermentation. Mucilage microbiology characterization and metataxonomic analysis were performed using 16S rDNA sequencing to determine bacterial diversity and ITS sequencing for fungal diversity. Additionally, the microorganisms were isolated into pure cultures. The molecular diversity analyses showed similarities in microorganisms present during the fermentation of Var. Castillo General and Var. Colombia, which are genetically closely related; mixed-acid bacteria (Enterobacteriaceae, Tatumella sp.) and lactic acid bacteria (Leuconostoc sp., Weissella sp. and Lactobacillaceae) were common and predominant, while in Var. Tabi, acetic acid bacteria (Gluconobacter sp. and Acetobacter sp.) and Leuconostoc sp. were predominant. At the end of the fermentation period, the fungi Saccharomycodaceae, Pichia and Wickerhamomyces were found in Var. Castillo General and Var. Colombia, while in Var. Tabi, Saccharomycodaceae, Pichia and Candida were recorded. Sensory analyses of the coffee beverages were carried out (SCA methodology) for all samples. Var. Tabi had the highest SCA score, between 82.7 and 83.2, while for Var. Colombia, the score ranged between 82.1 and 82.5. These three coffee varieties showed potential for the production of specialty coffees influenced by spontaneous fermentation processes that depend on microbial consortia rather than a single microorganism.

Microbial diversity on grape epidermis and wine volatile aroma in spontaneous fermentation comprehensively driven by geography, subregion, and variety

On their journey from the wine grape to the resulting wine, microbiota from grape surfaces controlled by multiple factors is transferred to wine spontaneous fermentation process with indisputable consequences for wine quality parameters. The associated microbiota was regionally distinct (defined to microbial terroir) but how these microbial patterns with significantly regional distinctiveness quantitatively drive the wine regional characteristics are not definite within a complete grape ecosystem at different geographical (> 300 km), subregional (< 10 km), and varietal scales. Here, we collected 24 samples (containing two grape varieties) from four subregions of two regions in Xinjiang wine production area to investigate fungal distribution patterns and the association with wine chemical composition at different evaluation scales. Meanwhile, the relationships were established between geographical, subregional, varietal community of fungi, and wine volatile aroma using partial least squares regression (PLSR) and structural equation modeling (SEM). Results show that microbial and volatile samples present the significantly regional difference inside the complete ecosystem. Microbiota showed a stronger heterogeneity at geography scales, which drove the distributions of subregional and varietal microbiota thereby influencing the volatile composition of finished wines. Moreover, geographical microbiota seems to weaken the effects of varietal community on wine aroma compounds. Microbial communities respond to environmental changes within a completely set grape-related ecosystem at different scales, and these responses resulted in the wine regional distinctiveness based on the volatile profiles. Our findings further confirmed the important role of microbial terroir in shaping wine styles and provided the new cerebration for the terroir drivers of microbiota.

Microbial communities and their correlation with flavor compound formation during the mechanized production of light-flavor Baijiu

Light-flavor Baijiu fermentation is a typical spontaneous solid-state fermentation process fueled by a variety of microorganisms. Mechanized processes have been increasingly employed in Baijiu production to replace traditional manual operation processes, however, the microbiological and physicochemical dynamics in mechanized processes remain largely unknown. Here, we investigated the microbial community succession and flavor compound formation during a whole mechanized fermentation process of light-flavor Baijiu using the conventional dilution plating method, PacBio single-molecule real-time (SMRT) sequencing and headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The results showed that largely different fungal and bacterial communities were involved in the soaking and fermentation processes. A clear succession from Pantoea agglomerans to Bacillus (B.) smithii and B. coagulans in dominant bacterial species and from Cladosporium exasperatum to Saccharomyces cerevisiae and Lichtheimia ramosa in dominant fungal species occurred in the soaking processes. In the fermentation process, the most dominant bacterial species was shifted from Pantoea agglomerans to Lactobacillus (La.) acetotolerans and the most dominant fungal species were shifted from Lichtheimia ramose and Rhizopus arrhizus to Saccharomyces cerevisiae. The bacterial and fungal species positively associated with acidity and the formation of ethanol and different flavor compounds were specified. The microbial species exhibited strong co-occurrence or co-exclusion relationships were also identified. The results are helpful for the improvement of mechanized fermentation process of light-flavor Baijiu production.

Microbial, physicochemical and proximate analysis of Tej collected from Amhara regional state of Ethiopia

Tej is an Ethiopian traditional alcoholic beverage with significant social and economic importance. Due to the spontaneous fermentation process of Tej, several issues such as safety, quality, and physicochemical properties of the final products is rquired to be assessed. Thus, this study was aimed to assess the microbial quality, physicochemical, and proximate properties of Tej associated with different maturity time. The microbial, physicochemical and proximate analyses were carried out by standard protocol. Lactic acid bacteria (6.30 log CFU/mL) and yeast (6.22 log CFU/mL) were the dominat microorganisms of all Tej samples at different maturity time, with significant differences (p = 0.001) in mean microbial count among samples. The mean pH, titratable acidity and ethanol content of Tej samples were 3.51, 0.79 and 11.04% (v/v), respectively. There were significant differences (p = 0.001) among the mean pH and titratable acidity values. The mean proximate compositions (%) of Tej samples were as follows: moisture (91.88), ash (0.65), protein (1.38), fat (0.47) and carbohydrate (3.91). Statistically significant differences (p = 0.001) were observed in proximate compositions of Tej samples from different maturity time. Generally, Tej maturity time has a great impact on the improvement of nutrient composition and the increment of the acidic contents which in turn suppress the growth of unwanted microorganisms. Further evaluation of the biological, and chemical safety and development of yeast-LAB starter culture are strongly recommended to improve Tej fermentation in Ethiopia.

New insights into the role of key microorganisms and wooden barrels during lambic beer fermentation and maturation

Belgian lambic beers are still produced through traditional craftsmanship. They rely on a spontaneous fermentation and maturation process that is entirely carried out in wooden barrels. The latter are used repetitively and may introduce some batch-to-batch variability. The present systematic and multiphasic study dealt with two parallel lambic beer productions carried out in nearly identical wooden barrels making use of the same cooled wort. It encompassed a microbiological and metabolomic approach. Further, a taxonomic classification and metagenome-assembled genome (MAG) investigation was based on shotgun metagenomics. These investigations provided new insights into the role of these wooden barrels and key microorganisms for this process. Indeed, besides their role in traditionality, the wooden barrels likely helped in establishing the stable microbial ecosystem of lambic beer fermentation and maturation by acting as an inoculation source of the necessary microorganisms, thereby minimizing batch-to-batch variations. They further provided a microaerobic environment, which aided in achieving the desirable succession of the different microbial communities for a successful lambic beer production process. Moreover, these conditions prevented excessive growth of acetic acid bacteria and, therefore, uncontrolled production of acetic acid and acetoin, which may lead to flavor deviations in lambic beer. Concerning the role of less studied key microorganisms for lambic beer production, it was shown that the Acetobacter lambici MAG contained several acid tolerance mechanisms toward the harsh environment of maturing lambic beer, whereas genes related to sucrose and maltose/maltooligosaccharide consumption and the glyoxylate shunt were absent. Further, a Pediococcus damnosus MAG possessed a gene encoding ferulic acid decarboxylase, possibly contributing to 4-vinyl compound production, as well as several genes, likely plasmid-based, related to hop resistance and biogenic amine production. Finally, contigs related to Dekkera bruxellensis and Brettanomyces custersianus did not possess genes involved in glycerol production, emphasizing the need for alternative external electron acceptors for redox balancing.

Advances in the Application of the Non-Conventional Yeast Pichia kudriavzevii in Food and Biotechnology Industries.

Pichia kudriavzevii is an emerging non-conventional yeast which has attracted increased attention for its application in food and biotechnology areas. It is widespread in various habitats and often occurs in the spontaneous fermentation process of traditional fermented foods and beverages. The contributions of P. kudriavzevii in degrading organic acid, releasing various hydrolase and flavor compounds, and displaying probiotic properties make it a promising starter culture in the food and feed industry. Moreover, its inherent characteristics, including high tolerance to extreme pH, high temperature, hyperosmotic stress and fermentation inhibitors, allow it the potential to address technical challenges in industrial applications. With the development of advanced genetic engineering tools and system biology techniques, P. kudriavzevii is becoming one of the most promising non-conventional yeasts. This paper systematically reviews the recent progress in the application of P. kudriavzevii to food fermentation, the feed industry, chemical biosynthesis, biocontrol and environmental engineering. In addition, safety issues and current challenges to its use are discussed.

Screening of Anti-fungal Bacillus Strains and Influence of their Application on Cocoa Beans Fermentation and Final Bean Quality

Aims: Contamination with filamentous fungi reduces the quality of cocoa beans and poses a health risk for consumers due to the potential accumulation of mycotoxins. The aim of this study was to develop antifungal Bacillus cultures for cocoa beans fermentation.&#x0D; Methods and Results: Antifungal activity of 7 Bacillus isolates was investigated using overlay technique. Solid culture media inoculated with bacterial and fungal isolates were cultivated under aerobic conditions at 30 ºC for 5 days. The Bacillus strain with strong antifungal activity was studied and inoculated to 18 kg box fermentations cocoa beans in Cote d'Ivoire in three time-independent replications each including a spontaneous control fermentation. The comparison of inoculated and spontaneous fermentation processes revealed that the Bacillus thuringiensis strain ATCC 10792 don't affected the fermentation process and cocoa bean quality. The anti-fungal in vivo assays revealed that the Bacillus thuringiensis strain ATCC 10792 completely inhibited growth of all fungi on the surface of cocoa beans. The result showed that sensory evaluation of chocolates is not significant differences for all treatment. The sensory evaluation of control and chocolates made from the inoculated cocoa beans with Bacillus showed excellent sensory quality, demonstrating that Bacillus thuringiensis strain ATCC 10792 did not affect aroma, texture and appearance of chocolate.&#x0D; Conclusion: From this study, it is revealed that the addition of Bacillus thuringiensis strain ATCC 10792 during fermentation cocoa beans suppress fungal growth and improve the quality of cocoa beans. Therefore, the anti-fungal culture Bacillus thuringiensis strain ATCC 10792 is recommended for future applications and its capacity to limit fungal growth during industrial-scale cocoa bean fermentation should be investigated in further studies.

Aroma precursor enhancing in dried cocoa beans fermentation using enzyme and heat addition

&lt;abstract&gt; &lt;p&gt;Changes in amino acids and reducing sugars in cocoa beans during fermentation were investigated using a 3 × 3 full complete factorial design using different enzyme additions (cellulase, papain and control–no enzyme) and water bath temperatures (40, 45 and 50 ℃) as variables over three days of fermentation. Aroma precursors (reducing sugars and free amino acids) developed inside the bean by enzymatic mechanisms during fermentation are converted into volatile compounds such as pyrazines and aldehydes during roasting. This study aimed to improve the fermentation process of dried beans by adding acetic acid, heat and enzymes, because there is insufficient pulp for the ideal spontaneous fermentation process. Samples were analyzed for fermentation index, cut bean, reducing sugar amino acid composition and volatile aroma composition profile using headspace solid phase microextraction (HS-SPME) and gas chromatography mass spectrometry (GC-MS). The results showed that the fermentation index was significantly affected by the addition of enzymes and water temperature. Although amino acids rose to 200%–300%, the composition contained several acidic amino acids because the pH utilized less than 4. Adding cellulase enzymes increases the amount of reducing sugars and amino acids but does not result in the formation of various amino acids.&lt;/p&gt; &lt;/abstract&gt;

Investigation by High-Throughput Sequencing Methods of Microbiota Dynamics in Spontaneous Fermentation of Abruzzo (South Italy) Wines

Spontaneous wine fermentation is a meaningful topic that cannot be disregarded among winemakers and consumers due to the peculiarity of the organoleptic profile that it confers to the wine. Nevertheless, in this process the activity of indigenous microorganisms might be a threat. We studied the evolution of the spontaneous fermentation process in a traditional Italian winery in order to understand the origin of spoilage microorganisms, and to characterize the peculiarity of the microbiota associated with spontaneous fermentation. Six Trebbiano and Montepulciano wine production chains were monitored by plate counts made by OIV methods and by Illumina MiSeq technique. Despite some compositional deficiencies, all grape musts were characterized by a highly concentrated microbial population. Non-Saccharomyces yeasts revealed an unexpected tolerance to ethanol, which has contributed to the evolution of alcoholic fermentation. Lactic bacteria were detectable from the very first steps of the winemaking process, with a prevalence of Leuconostoc spp. which is nowadays, rarely isolated in wine. The combination between culture-dependent and high-throughput sequencing (HTS) approaches allowed to estimate microbial diversity and growth dynamics in wine fermentations of different grape varieties and under different treatments; these results could be used by winemakers as a starting point to drive a more mindful, accurate and, controlled fermentation process and to set up the most suitable environmental conditions to enhance wine singularities.

Sustainable Production and Characteristics of Dried Fermented Vegetables

The current fashion for healthy food and the increasing number of people with lactose intolerance make fermented vegetables increasingly important. On top of this, surpluses unused in the vegetable harvest can become a potential source of “green waste”. The use of fermentation and freeze-drying can result in a valuable, sustainable product that can solve the problems of spoiled vegetables and the need for refrigerated storage. Therefore, this study aimed to obtain sustainable dried fermented vegetables and to compare their selected physical and structural properties. Beetroot, carrot, and red pepper were selected for this purpose. These vegetables were subjected to a spontaneous lactic fermentation process. After the process, the vegetables were freeze-dried, and their structure and selected properties (color, dry weight, and the number of lactic acid bacteria) were determined. Fermented vegetables were found to differ from their raw sources in structure and color, the main discrepancies being shown by the b* factor (yellow-blue). Root vegetables had smaller pores of structure in the freeze-dried samples than red peppers. The freeze-drying process did not affect the number of bacteria. It can be concluded that both the fermentation and the freeze-drying processes affected the structure of the selected vegetables. All tested vegetables can be fermented and freeze-dried without major changes in color and microbiological properties and can be used as a potential source of lactic acid bacteria and health-promoting pigments, e.g., in the form of chips. In addition, their shelf life is extended.

Autochthonous starter culture selection for Salame Piemonte PGI production

Salame Piemonte is a dry-fermented meat product typical of the Piedmont region in Italy, manufactured using commercial starter cultures. This study aimed to select autochthonous starter cultures (ASCs) that could be used for sausage fermentation in order to strengthen the link with the geographical area of production and improve the sensory properties of the final product. A culture-dependent approach was adopted during three different spontaneous sausage fermentation processes to isolate and characterise the main bacterial resources involved. Dominant lactic acid bacteria (LAB) in each batch were Pediococcus pentosaceus, Latilactobacillus sakei, and Latilactobacillus curvatus; Staphylococcus xylosus was the most dominant coagulase-negative staphylococci (CNS) in all the studied batches. LAB and presumptive CNS isolates were further evaluated for their physiological properties and biotechnological potential. Thereafter, 11 strains were selected and evaluated for safety. Five selected strains (two P. pentosaceus, two L. sakei, and one S. xylosus strain) were used for pilot-scale Salame Piemonte production with seven different strain combinations. Based on the liking test, three ASC combinations led to the highest liking score compared to industrial products. These three ASCs were then used for the second pilot-scale sausage production confirming the high liking score. In summary, the use of P. pentosaceus and S. xylosus ASC significantly improved product sensory properties compared with that obtained using commercial starter cultures.

Microbial and Chemical Dynamics during Marula Wine Fermentation

Marula wine is traditionally produced through a spontaneous fermentation process and has a huge economic potential in Africa. The current study investigated the contributing microbiota and the metabolites produced during the wine fermentation process. Microbial communities were analyzed by selective cultivation and identified by biotyping and rDNA sequencing. Sugars and volatile compounds were determined with the high performance liquid chromatography and gas chromatography, respectively. Different Lactobacillus spp. were present throughout the fermentation process but dominated the earlier stages of fermentation, together with non-Saccharomyces yeasts, whereas Saccharomyces cerevisiae and acetic acid bacteria dominated the latter stages. Sucrose, glucose and fructose were detected during the early stages, while ethanol and butanol were present during the latter stages of fermentation. Interestingly, acetic acid and formic acid were detected in relatively high amounts at the latter stages of fermentation. Lactobacillus spp. and S. cerevisiae were identified as the primary contributing microbiota, and Acetobacter aceti and Acetobacter pasteuriannus were associated with the off taste and spoilage of the marula wine.

Kinetics Crystallization and Polymorphism of Cocoa Butter throughout the Spontaneous Fermentation Process.

The spontaneous fermentation process of Criollo cocoa is studied for its importance in the development of chocolate aroma precursors. This research supports the importance of spontaneous fermentation, which was studied through the crystallization behavior and polymorphisms of cocoa butter (CB), the most abundant component of chocolate that is responsible for its quality physical properties. The k-means technique was used with the CB crystallization kinetics parameters to observe the division of the process during the first stage (day 0–3). The experimental crystallization time was 15.78 min and the second stage (day 4–7) was 17.88 min. The Avrami index (1.2–2.94) showed that the CB crystallizes in the form of a rod/needle/fiber or plate throughout the process. CB produced metastable crystals of polyforms and . Three days of fermentation are proposed to generate Criollo cocoa beans with acceptable CB crystallization times.