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High-yield resistant hybrids are used in cocoa fermentation and result in chocolates with different sensorial profiles. This work aimed to characterize the fermentation microbiologically and physicochemically. Hybrids CEPEC 2004, FA13, PH15, and CEPEC 2002 were used for fermentation. The yeast, acetic acid bacteria, lactic acid bacteria, and mesophilic bacteria population were evaluated in their respective medium. Carbohydrates and acids were detected using a high-performance liquid chromatography system, and volatiles were analyzed using gas chromatography-mass spectrometry equipment. Finally, a consumer acceptance test followed by a check-all-that-apply question and a temporal dominance of sensations assessment was performed in chocolate. The fermentation resulted in a typical succession: yeast-dominated at first, followed by lactic acid, acetic acid, and mesophilic bacteria. In the pulp, carbohydrates and citric acid were consumed. Low concentrations of acetic acid (0.09-1.75 g/kg) were detected. Acids, esters, and alcohols were the most abundant groups. The chocolate profile resulted in sweet, acidic, and fruity, satisfying consumers' tastes. PRACTICAL APPLICATION: The cocoa hybrid-mix fermentation can improve the fermentation process and chocolate quality. The mixture generated a different sensory profile in comparison to other fermentations. The fruity chocolate was accepted and liked by consumers.

We evaluated the probiotic properties of lactic acid bacteria using resistance, safety, and functional assays. A preliminary subtractive screening of nineteen strains was performed based on their survival in simulated gastric and intestinal juice, and cell surface characteristics (hydrophobicity and auto-aggregation). Five strains were selected for further characterization, which included the assessment of their co-aggregation to pathogens, phenol tolerance, antimicrobial activity, and safety. Moreover, their adhesion to Caco-2 and HT-29 cells and the ability to inhibit pathogenic bacteria adhesion were evaluated. All strains had high (≥ 80.0%) survival rates in gastric and intestinal juices. Among them, Lactobacillus brevis CCMA 1284, L. plantarum CCMA 0743, and L. plantarum CCMA 0359 exhibited higher hydrophobicity (95.33, 96.06, and 80.02%, respectively), while L. paracasei CCMA 0504 and L. paracasei CCMA 0505 had the highest auto-aggregation values (45.36 and 52.66%, respectively). However, these last two strains were positive for the DNAse test, which is a safety concern. The CCMA 0359 and CCMA 1284 strains did not show antimicrobial activity, while the CCMA 0505 strain had a higher percentage of adhesion (4.75%) to Caco-2 cells. In the simulated competition and exclusion assays, the CCMA 0743 strain was able to reduce Salmonella adhesion to both cells (Caco-2 and HT-29), but only the CCMA 0743 and CCMA 0505 strains inhibited Escherichia coli adhesion to HT-29 cells in the competition assay. According to the results of these evaluated attributes, this strain showed to be an excellent candidate for probiotic use.

The Saccharomyces cerevisiae has been used for many years in the elaboration of food and beverage products, mainly associated with fermentation processes. The objective of this study was to characterize different indigenous S. cerevisiae strains and guide the notable strains for potential use in productions of fermented maize-based beverages. Initially, 81 strains isolated from different spontaneous food fermentations were evaluated. About 31% of strains showed phytase activity, an important characteristic for their application in cereals beverages production. All strains were able to grow in pH values 2.0, 3.0, and 5.0 and the presence of 5, 15, and 30% of glucose, but none could grow at 42 °C. Only 29.6% of the evaluated strains were able to efficiently grow in up to 1.0 mol L(−1) of NaCl. The Rep-PCR and RAPD-PCR tools showed that the strains were differently grouped by the two techniques, and the grouping was not completely correlated with isolation source. A total of 65 volatile compounds were identified from the maize beverage produced. The profiles of volatile compounds produced by the strains were strain specific. S. cerevisiae strains isolated from the same source showed different chemical and genetic profiles, emphasize the importance to characterize the performance of each strain when searching for starter culture to develop or improve fermented beverages. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s42770-020-00271-8) contains supplementary material, which is available to authorized users.

Fruit juices have shown promising results as new probiotic carriers. This study aimed to evaluate acerola, jelly palm, and passion fruit juices as substrates for fermentation using Lactiplantibacillus plantarum CCMA 0743 and Lacticaseibacillus paracasei LBC-81 in single and mixed cultures. First, the juices were evaluated as substrate and selected based on bacterial growth performance during fermentation. Afterward, the impact of fermentation on sugars, organic acids, and bioactive compounds was also appraised. Phytochemical modification of three different juices fermented by lactic acid bacteria at 37 °C/24 h was evaluated. After 18 h of fermentation, passion fruit juice showed higher cell viable counts of single and mixed L. plantarum CCMA 0743 culture, above 9.00 Log CFU/mL, and pH between 4.07 and 4.10. Sugars consumption and organic acid production were influenced by juice composition and culture used. The mixed culture reduced the total sugars in the passion fruit juice by approximately 53.0% (8.51 g/L). Lactic acid was the main product of the sugars fermentation, with higher concentrations detected in passion fruit juice (8.39-11.23 g/L). Bioactive compounds were analyzed on the selected substrate. The fermentative process reduced antioxidant activity and carotenoid content. However, single L. plantarum CCMA 0743 culture increased the yellow flavonoid content of passion fruit juice by approximately 3.0 µg/mL. L. plantarum CCMA 0743 showed high and suitable cell, viable counts, to claimed probiotic products, increasing bioactive compounds in passion fruit juice. Therefore, this strain and passion fruit substrate showed attractive potential to produce alternative and functional fermented fruit beverages.

This work aimed to evaluate the performance of co-cultivation of potential probiotic yeast and lactic acid bacteria (LAB) in producing plant-based fermented beverages. The co-culture comprised LAB Lactiplantibacillus plantarum CCMA0743 with the yeasts Pichia kluyveri CCMA 0615, Pichia guilliermondii CCMA 1753 and Debaryomyces hansenii CCMA 1761 separately. The plant substrate was 75 g oat, 175 g sunflower seeds, and 75 g almonds. The viability of microorganisms in the plant-based matrix was evaluated during fermentation, storage at 4 °C, and under simulated gastrointestinal tract (GIT) conditions. Chemical analysis, antioxidant activity, and sensory profile of the beverages were also determined. The three yeasts and the LAB showed counts greater than 6.0 log CFU/mL after fermentation, and the plant-based matrix protected the yeasts during simulated digestion. P. kluyveri and D. hansenii showed higher survival than P. guilliermondii and L. plantarum after exposure to simulated GIT conditions. The pH of the plant-based matrix reduced from approximately 7 to 3.8. Lactic acid was the main organic acid produced during fermentation. In addition, 113 volatile compounds were detected by gas chromatography-mass spectrometry (GC-MS), including alcohols, aldehydes, alkanes, alkenes, acids, ester, ether, ketones, phenol, and amides. The beverage sensory profile varied with the co-culture. The co-culture D. hansenii and L. plantarum showed higher antioxidant activity than the other co-culture tested, and the homogeneous texture attribute characterized the beverage produced with this combination. Results show the suitability of tested co-cultures to produce a plant-based fermented beverage and indicate more significant potential for D. hansenii and L. plantarum co-culture as a starter for its functionalization.

The probiotic strains Lactobacillus brevis CCMA1284 and Lactobacillus plantarum CCMA0359 were microencapsulated by spray drying using different matrices – whey powder (W), whey powder with inulin (WI) and whey powder with maltodextrin (WM). Viability of the microencapsulated strains in acid and bile juices and during 90 days of storage (seven and 25 °C) was evaluated. The two strains exhibited high encapsulation efficiency (> 86%) by spray drying. The different matrices maintained L. plantarum viability above six log CFU g⁻¹ at 7 °C for 90 days, whereas similar results for L. brevis were observed only for W. The use of inulin as matrix of encapsulation did not enhance bacterial viability in the evaluated conditions. In general, the use of W and WM as matrices was effective for L. plantarum viability. However, only W was effective for L. brevis in the evaluated conditions. The spray drying technique was successfully adopted for the encapsulation of L. plantarum CCMA0359 and L. brevis CCMA1284 strains.

Background The demand for industrialized foods that contribute to health and well‐being has characterized the new generation of consumers. Yam (Dioscorea sp.) is a nutritious food; however, it is not used very much in industrial food processes. The objective of this study was to develop and to characterize a truly dairy‐free low‐fat ice cream prepared from unfermented and fermented with yam dough. Results The fermentation was conducted by Leuconostoc lactic CCMA 0415 remained viable (107 CFU/g) during 90 days of storage. The fermentation process reduced the starch concentration from 26.82% to 22.35% and the protein concentration from 4.68% to 3.99% and increased the concentration of some minerals (K, S, Cu, Mn, Zn, and Fe). The total phenolic contents for fermented and unfermented ice creams were 51 and 54 mg, respectively. The radical scavenging activity were 18% and 10% with the 1,1‐diphenyl‐2‐picrylhydrazyl method and 44% and 26% with the 2,2’‐azino‐bis (3 ethylbenzo‐thiazoline‐6‐sulfonic acid) method for the unfermented and fermented samples, respectively. The fermented and unfermented ice creams were both characterized as non‐Newtonian fluids exhibiting pseudoplastic behaviors. Conclusions These results indicated that yam is a suitable substitute for milk, thus making it an option to produce edible lactose‐free ice cream with low fat.

While many bacteria have been used as probiotics by industries, only two yeasts, Saccharomyces cerevisiae var. boulardii and Kluyveromyces fragilis (B0399), have been used for this purpose. In the present work, a total of 116 yeasts isolated from Brazilian indigenous fermented food, cocoa fermentation, and kefir were in vitro characterized for probiotic attributes. From 116 isolates, 36 were tolerant to gastrointestinal conditions evaluated by tolerance to pH 2.0, bile salts (0.3% w/v), and 37 °C temperature. From those, 15 isolates showed a similar or higher percentage (P 0.05) of hydrophobicity, autoaggregation, and coaggregation with E. coli than the reference strain S. boulardii. All these strains showed a high percentage of adhesion to Caco-2 cells ( 63%) and antioxidant activity (ranging from 18 to 62%). Phytate hydrolysis was evaluated for these yeasts and 13 strains showed positive results, which is important for nutrient availability in plant-based foods. These results are important insights for characterization of novel probiotic yeast strains as well as to aggregate functional value to these food products.

To reduce postharvest losses, substandard fruit and agricultural surpluses can productively be used as raw material for vinegar production. The present study aimed to prepare vinegars from surpluses of physalis (Physalis pubescens L.) and red pitahaya (Hylocereus monacanthus) and then evaluate their sensorial characteristics, antimicrobial activities, total phenolic content (TPC) and total antioxidant capacity by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2?-azino-bis(3-ethylbenzo thiazoline-6-sulfonic acid) methods. RESULTS Two vinegars were produced by submerged fermentation using physalis and red pitahaya fruits surpluses. Physalis and red pitahaya vinegars had 47 and 45 g L?1 acetic acid, respectively, and both vinegars contained approximately 1 g L?1 ethanol. Both vinegars displayed antimicrobial activity against Escherichia coli, Listeria monocytogenes, Staphylococcus aureus and Salmonella enteritidis. The TPC of physalis and red pitahaya vinegar was 0.5638 and 0.3656 g L?1 g gallic acid L?1 for physalis and red pitahaya, respectively. A similar antioxidant activity was detected in both the wines and vinegars. The sensorial analysis revealed that the consumers ?like moderately? each vinegar, and citric aroma was noted in the physalis vinegar. CONCLUSION In the present study, vinegars with sensorial characteristics approved by consumers were developed using fruit surpluses, adding value through a new product making use of a simple methodology that is both inexpensive and demonstrates a good yield. ? 2018 Society of Chemical Industry

Several non-dairy probiotic beverages are already available to consumers and have been considered suitable carriers for probiotic bacteria. This study aimed to investigate the effect of Lactiplantibacillus plantarum CCMA 0743 in single and co-culture on the volatile compounds and sensory profiles of fermented passion fruit juice. The viability of strains inoculated in juice and MRS matrices was evaluated in a simulated gastrointestinal condition. The bacterial viability after 28 days of refrigerated storage of the juices was also evaluated. L. plantarum CCMA 0743 showed high viability (6.18 Log CFU/mL) after passage throughout simulated digestion in the passion fruit juice matrix. Both juices maintained high probiotic counts (>8.0 Log CFU/mL) during storage. Also, the yellow color was stable after 28 days of storage. Volatile compounds of passion fruit juices were modified after the fermentation process, such as ketones and alcohol formation degradation. The sensory profile of passion fruit juice was modified by single and co-culture fermentations. The fermented samples were mainly correlated with the terminologies “salty, acidic and bitter tastes” and “sweetener aftertaste”. Overall, passion fruit juice proved to be an adequate food matrix to deliver the evaluated strains. However, individual strains or strain-strain interactions with the food matrix affect the fermented product, demonstrating that strain and matrices evaluations are essential for developing novel products with acceptable characteristics.

Cereal-based fermented beverages are non-dairy products which are considered possible carriers for probiotic strains and alternatives for use by vegans and lactose-intolerant consumers. In the present work, the commercial probiotic, Lactobacillus paracasei LBC-81, was used singly and in co-culture with potential probiotic yeasts, Saccharomyces cerevisiae CCMA 0731, S. cerevisiae CCMA 0732, and Pichia kluyveri CCMA 0615, to ferment a maize-based substrate. All tested strains showed viability higher than 6 log CFU/mL, as recommended for food probiotic products, except for the yeast P. kluyveri which decreased during fermentation and storage time. A reduction in pH value, from approximately 7 to 4, was observed. This decrease was due organic acid production, which did not affect the microbial viability. Lactic and acetic acids were the main organic acids produced during fermentation, and they decreased over 28 days of storage (0.5 and 0.1 g/L for lactic and acetic acids, respectively). Ethanol was detected in the S. cerevisiae assays; however, the content was5 g/L in a non-alcoholic beverage. Seventy volatile compounds were detected, including acids, alcohols, aldehydes, esters, ketones, and other compounds. Sensory analysis showed score of 5.93-4.57, respectively for appearance and taste. This is an important result, considering that the beverage had no flavoring additive and lacked a sweet taste. Therefore, probiotic beverages were successfully obtained by maize fermentation inoculated with co-culture of S. cerevisiae (CCMA 0731 or CCMA 0732) and L. paracasei LBC-81.

The survival of bacterial strains isolated from tejuino was evaluated in the human gastrointestinal model (ARIS) using saline solution and simulated food as matrices. Antagonistic activity towards pathogenic bacteria, short-chain fatty acids production and adhesion to HT-29 cells were also studied. Leuconostoc citreum CIATEJ BI–49.1 , Weissella cibaria CIATEJ BI–48.1 , Chryseobacterium bovis CIATEJ BI–55.1 and Bacillus safensis CIATEJ BI–75.1 were selected and inoculated in ARIS model. B. safensis and C. bovis were unable to survive the simulated human gastrointestinal tract conditions. Leu. citreum seemed to be more resistant to gastrointestinal conditions than W. cibaria , recovering 7–8 log CFU/mL. In the antagonistic assay, Leu. citreum showed the highest (p S. aureus and S. Typhimurium strains, while W. cibaria had the highest (p L. monocytogenes . In regards of short chain fatty acids production, Leu. citreum showed highest (p W. cibaria (approximately 0.5%). Leu. citreum could be a potential probiotic strain due to its tolerance to gastrointestinal conditions, antagonistic activity towards foodborne pathogens, short-chain fatty acids production and adhesion to HT-29 cells.

This work aimed to develop a nondairy fermented beverage from a blend of cassava and rice based on Brazilian indigenous beverage cauim using probiotic lactic acid bacteria (LAB) and yeast. The indigenous strains Lactobacillus plantarum CCMA 0743 (from cauim) and Torulaspora delbrueckii CCMA 0235 (from tarubá), and the commercial probiotic, L. acidophilus LAC-04, were used as starter cultures in single and co-cultivations. The bacteria populations were around 8.0 log (CFU/mL) at the end of all fermentations as recommended for probiotic products. Higher residual starch contents were noted in the single LAB cultures (10.6% [w/w]) than in co-cultures (6% [w/w]), showing that co-culture may help the digestibility. For all different assays (single and co-culture), lactic acid was the main organic acid detected (1.6g/L) and ethanol was lower than 0.5% (w/v) consisting in a non-alcoholic beverage. The assays containing yeast showed the highest antioxidant activity (around 10% by DPPH and ABTS methods). Therefore, a nondairy fermented beverage was successfully obtained, and the co-culture of LAB and T. delbrueckii could increase the product's functional properties.

The main carbon source used for growth by four yeast strains (Yarrowia lipolytica CCMA 0357, Y. lipolytica CCMA 0242, Wickerhamomyces anomalus CCMA 0358, and Cryptococcus humicola CCMA 0346) and their lipid production were evaluated, using different concentrations of crude and pure glycerol and glucose. Whereas crude glycerol (100 g/L) was the main carbon source used by Y. lipolytica CCMA 0357 (nearly 15 g/L consumed at 120 hr) and W. anomalus CCMA 0358 (nearly 45.10 g/L consumed at 48 hr), pure glycerol (150 g/L) was the main one used by C. humicola CCMA 0346 (nearly 130 g/L consumed). On the other hand, Y. lipolytica CCMA 0242 used glucose (100 g/L) as its main source of carbon (nearly 96.48 g/L consumed). Y. lipolytica CCMA 0357 demonstrated the highest lipid production [about 70% (wt/wt)], forming palmitic (45.73% of fatty acid composition), stearic (16.43%), palmitoleic (13.29%), linolenic (10.77%), heptadecanoic (4.07%), and linoleic (14.14%) acids. Linoleic acid, an essential fatty acid, was produced by all four yeast strains but in varying degrees, representing 70.42% of the fatty acid profile of lipids produced by C. humicola CCMA 0346.

Microbial diversity, final concentration of volatiles compounds during fermentation and the characterization and acceptability of chocolates of three cocoa hybrids (PH 9, PH 15 and PH 16) were evaluated. Hanseniaspora uvarum, Pichia kluyveri, Pichia caribbica and Saccharomyces cerevisiae were the predominant yeasts in the three fermentations. Lactobacillus plantarum, Acetobacter pasteurianus, Bacillus cereus and Lysinibacillus fusiformis were the common bacterial species isolated from the fermentations. The PH 9 hybrid showed highest diversity of lactic acid bacteria and acetic acid bacteria. Twenty-seven volatile compounds were identified during the cocoa fermentation of all hybrids. Chocolate from PH 9 hybrid had most desirable flavors and obtained the most ideal score in acceptance test. Chocolate from PH 15 hybrid was considered the most bitter, acidic and astringent. Chocolates produced by different cocoa hybrids showed differences in sensory attributes and acceptance. Therefore, special management is required for the different hybrid fermentations to obtain high-quality chocolate. Practical Applications As a solution to the recovery of the cocoa crop in Brazil, many hybrids resistant to the disease called “witches’ broom” were developed, but the impact of these crosses on cocoa quality, consequently in chocolate is not known. The producers’ cooperatives and farmers of cocoa have no technical or scientific information about the behavior of cocoa hybrids during spontaneous fermentation process, which, oftentimes give rise to an uncontrolled fermentation of cocoa hybrids mixture. The results of this paper could be used among the producers’ cooperatives and cocoa farmers for guidance about the influence of genetic improvement on the quality of cocoa fermented beans and chocolates, particularly among the three hybrids evaluated in this study.

The total antioxidant capacity (TAC) and total phenolic compounds (TPC) of cocoa beans and chocolate produced from spontaneous and inoculated fermentations of different cocoa varieties were evaluated. Fourier transform infrared spectroscopy (FTIR), as well as conventional methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), was used to determine TAC and TPC. Chocolate showed higher (p0.05) TPC (47.17-57.16mgGAE/g) and TAC (1.66-2.33mMTE/g and 8.86-11.35mMTE/g as measured by DPPH and ABTS, respectively) than cocoa beans (6.30-26.05mgGAE/g, 0.24-1.17mMTE/g and 1.29-4.83mMTE/g for TPC, DPPH and ABTS, respectively). Partial least square (PLS) model for infrared data showed a good calibration coefficient (R

The use of lactic bacteria in the development of functional foods has increased in recent years. In addition to their probiotic characteristics, they can ferment a variety of substrates, such as cereals, roots, and tubers. Phytase producer lactic acid bacteria strains and their behavior during the fermentation process of yam-based food were studied. Leuconostoc lactis CCMA 0415, Lactobacillus plantarum CCMA 0744, and Lactobacillus fermentum CCMA 0745 were selected due to phytase production, pH reduction, and growth during 24 h of fermentation. Oxalate activity was not detected in all assays, suggesting its concentration was reduced due to the bleaching process. Among the selected strains, L. lactis CCMA 0415 appeared to be a promising strain in yam-based fermentations because it maintained a cell viability above 8 log CFU/mL and did not reduce diosgenin concentrations (around 8.0 μg/mL) after fermentation for 24 h, thereby, generating a potentially functional yam food. Furthermore, this strain promoted the decrease of pH value from 6.1 to 3.8 and produced 8.1 g/L lactic acid, at 6 h of fermentation. The L. lactis CCMA 0415 was reported as a starter culture in fermented products based on cereals, roots, and tubers.

The development of nondairy fermented beverages with health benefits has been a key for what is termed the functional food market. Cereal grains are one of the most important sources of protein, carbohydrates, vitamins, minerals, and fiber for the worldwide consumers. Traditional fermented foods and beverages still play a significant role in the diet of numerous civilizations, and the world dietary culture can be classified based on the staple cereal used. In the last few decades, the literature regarding the technological and nutritional aspects of fermented beverages has increased substantially. Consequently, new insights about microorganisms and bioactive compounds have improved, helping develop novel fermented beverages with healthy properties. This paper will assess the relevant findings concerning the characteristics of traditional fermented beverages produced from cereals by focusing on their functional properties. Furthermore, the efforts to improve the health benefits of these beverages will be explored.

Properly conducted cocoa fermentation is an important step for the production of high-quality chocolate. The aim of this work was to investigate the effect of four cocoa varieties (CCN51, PS1030, FA13, and CEPEC 2004) inoculated with Saccharomyces cerevisiae CA11 on microbial communities and the profile of volatile compounds and sensory characteristics of chocolate. The S. cerevisiae population increased significantly (p S. cerevisiae and Hanseniaspora guilliermondii , while Lactobacillus casei and Gluconobacter oxidans were the predominant bacteria in the four different fermentations analyzed. Sixty-one volatile compounds—including aldehydes (11), ketones (10), esters (14), acids (8), alcohols (8), pyrazines (5), furans (3), and others (caffeine and heptadecane)—were detected and quantified by GC–MS in the different chocolates. The sensory analysis showed that caramel was perceptible in the chocolate of PS1030, while CEPEC2004 was related to astringency, bitterness, and chocolate flavor attributes. The chocolates produced from FA13 and CCN51 were more similar in terms of sour and chocolate aroma. A “temporal dominance of sensation” (TDS) analysis showed that although the bitter attribute was dominant, the fruity, sweet, sour, astringent, and cocoa attributes were also perceptible, depending on the cocoa variety. These results suggest that the cocoa varieties had an influence on the chocolate's quality, which should be considered to obtain chocolate with different sensory characteristics or for better standardization of the process, even when using yeast as a starter culture.

The consumption of probiotics has increased due to the reported health benefits, mainly in preventing or treating gastrointestinal pathology. This study investigated the antimicrobial capacity of yeasts, Saccharomyces cerevisiae and Pichia kluyveri, previously isolated from fermented foods (indigenous beverage, kefir and cocoa) against the adhesion of foodborne pathogens to Caco-2 cells. Co-aggregation of yeasts with pathogens and were evaluated by quantitative analysis and using scanning electron and laser confocal microscopies. All yeasts strains were able to co-aggregate with the tested pathogens, however, this activity was strain-dependent. The inhibition tests showed that the adhesion of Escherichia coli EPEC, Listeria monocytogenes and Salmonella Enteritidis to Caco-2 was reduced by all the yeasts studied. Most of the evaluated yeasts showed inhibition rates equal to or greater than the commercial probiotic Saccharomyces boulardii. The yeasts were able to reduce up to 50% of the bacterial infection, as observed for CCMA0615 towards EPEC in exclusion assay; CCMA0731, CCMA0732 and CCMA0615 towards L. monocytogenes in exclusion and competition assays; and CCMA0731 in exclusion and CCMA0731, CCMA0732, CCMA0615 in competition assay towards S. Enteritidis. No antimicrobial compounds were produced by the yeasts, showing that competition for nutrients and/or receptors in the intestinal mucosa was the mechanism to bacterial inhibition.

Traditional Brazilian indigenous fermented foods and beverages are potential sources of new food products that promote health, but they are still produced by natural fermentation. In the present work, Lactobacillus fermentum CCMA 0215 isolated from the indigenous fermented cassava beverage yakupa was used as single or mixed starter culture with five different yeast strains (Torulaspora delbrueckii CCMA 0234 and CCMA 0235, Pichia caribbica CCMA 0198, and Saccharomyces cerevisiae CCMA 0232 and CCMA 0233) to ferment cassava. Fermentations using each yeast as single starter culture were also performed. The microbial population and metabolites produced during cassava fermentation were investigated. In all assays, the inoculated microorganisms fermented cassava, judged by lowering the pH from 6.0 to 4.0–5.0 within 24h. Lactic acid bacteria (LAB) and yeast population increased during fermentation. Lactic acid was the main organic acid produced, reaching a maximum value of 4.5g/L at 24h in the co-culture with L. fermentum CCMA 0215 and T. delbrueckii CCMA 0234. Other organic acids, such as malic, tartaric, and succinic acids, were detected in low concentrations (less than 0.5g/L). Ethanol and glycerol were produced in all assays inoculated with yeasts (single and co-cultured with LAB), reaching the maximum concentration of approximately 2.3g/L and 0.6g/L, respectively. Twenty-two volatile compounds were detected after 48h of fermentation, varying widely between single and co-cultures. The compounds 2-phenylethyl alcohol, 1-butanol, 3-methyl (isoamyl alcohol), and acetoin were detected in single and co-cultures. This study demonstrated co-cultures of yeasts and LAB had the ability to improve the aroma profile of the final product and the safety of the product by lowering the pH.

The dynamic of Saccharomyces cerevisiae, Pichia kluyveri and Hanseniaspora uvarum during spontaneous and inoculated cocoa fermentations and their effect on sensory characteristics of chocolate were investigated. Yeast populations were assessed by qPCR. S. cerevisiae was predominant during spontaneous (average 5.4 log cell/g) and inoculated (average 7.2 log cell/g) fermentations. The H. uvarum seemed to be suppressed by the other two yeasts, as it showed similar population (approximately 4.0 log cell/g) even in the inoculated assay. Carbohydrates were consumed quickly at inoculated fermentation (68% and 42% were consumed in the inoculated and control assays respectively, at 24 h). Ethanol content was higher in the inoculated (8.3 g/kg at 48 h) than in the control (4.6 g/kg at 96 h) fermentation. Consumers did not report a significant preference for either chocolate (p

The aim of this work was to identify and characterize the microbiota present during fermentation and in the final beverage, tarubá, by culture-dependent and -independent methods. In addition, target chemical compounds (carbohydrates, organic acids, and ethanol) were evaluated. Lactic acid bacteria (LAB) and mesophilic bacteria were the predominant microorganisms. Among them, Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides, and Bacillus subtilis were frequently isolated and detected by DGGE analysis. Torulaspora delbrueckii was the dominant yeast species. Yeast isolates Pichia exigua, Candida rugosa, T. delbrueckii, Candida tropicalis, Pichia kudriavzevii, Wickerhamomyces anomalus, and Candida ethanolica and bacteria isolates Lb. plantarum, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus sp., and Chitinophaga terrae showed amylolytic activity. Only isolates of P. exigua and T. delbrueckii and all species of the genus Bacillus identified in this work exhibited proteolytic activity. All microbial isolates grew at 38 °C, and only the isolates belonging to Hanseniaspora uvarum species did not grow at 42 °C. These characteristics are important for further development of starter cultures; isolates of T. delbrueckii, P. exigua, and Bacillus species identified in this work displayed all of these properties and are potential strains for use as starter culture in cassava fermented food.

This exploratory work aims to identify the potential inhibitors of lactic bacterial growth and to propose enological practices to guarantee the occurrence of spontaneous malolactic fermentation (MLF) in wines from traditional and double-pruning management harvests in southeast Brazil. One white wine from a summer harvest and one red wine from a winter harvest that failed to complete MLF were utilized as comparative models to identify inhibitor compounds to lactic bacteria. Wine composition, alcoholic-fermentation temperature and bacterial strain contribute to the success or failure of MLF. Temperatures below 12 °C during alcoholic fermentation decrease lactic bacterial metabolism and may impair the bacteria’s growth after yeast cells lysis. A must pH below 3.2 in a summer harvest impairs bacterial growth, and the association of low pH with a free-SO2 concentration above 10 mg L-1 may inhibit MLF. For grapes with a high sugar content, harvested in the winter cycle, enologists should keep the alcohol content below 15% and control the alcoholic-fermentation temperature.

The microbial community of artisanal corn fermentation called Chicha were isolated, purified and then identified using protein profile by Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and confirmed by partial ribosomal gene sequencing. Samples from Chicha beverage were chemically characterized by gas and liquid chromatography (HPLC and GC-MS). Aerobic mesophilic bacteria (AMB) (35.8% of total of isolated microorganisms), lactic acid bacteria (LAB) (21.6%) and yeast (42.6%) were identified. Species of the genera Klebsiella, Bacillus, Staphylococcus, Micrococcus, Enterobacter, and Weissella were identified. Rhodotorula mucilaginosa, Lodderomyces elongisporus, Candida metapsilosis, and C. bohicensis were the yeasts found. The LAB isolates detected were responsible for the high concentrations of lactic acid found during the fermentation process (1.2 g L− 1), which is directly related to the decrease in pH values (from 6.95 to 3.70). Maltose was the main carbohydrate detected during corn fermentation (7.02 g L− 1 with 36 h of fermentation). Ethanol was found in low concentrations (average 0.181 g L− 1), making it possible to characterize the beverage as non-alcoholic. Twelve volatile compounds were identified by gas chromatography; belonging to the groups acids, alcohols aldehydes, acetate and others. MALDI-TOF was successfully used for identification of microbiota. Weissella confusa and W. cibaria were detected in the final product (after 36 h of fermentation), W. confusa is often classified as probiotic and deserve further application studies.

In the present work, an in vitro model of the gastrointestinal tract (GIT) was developed to obtain real-time observations of the pH homeostasis of single cells of probiotic Lactobacillus spp. strains as a measure of their physiological state. Changes in the intracellular pH (pHi) were determined using fluorescence ratio imaging microscopy (FRIM) for potential probiotic strains of Lactobacillus plantarum UFLA CH3 and Lactobacillus brevis UFLA FFC199. Heterogeneous populations were observed, with pHi values ranging from 6.5 to 7.5, 3.5 to 5.6 and 6.5 to 8.0 or higher during passage of saliva (pH 6.4), gastric (pH 3.5) and intestinal juices (pH 6.4), respectively. When nutrients were added to gastric juice, the isolate L. brevis significantly decreased its pHi closer to the extracellular pH (pHex) than in gastric juice without nutrients. This was not the case for L. plantarum. This study is the first to produce an in vitro GIT model enabling real-time monitoring of pH homeostasis of single cells in response to the wide range of pHex of the GIT. Furthermore, it was possible to observe the heterogeneous response of single cells. The technique can be used to determine the survival and physiological conditions of potential probiotics and other microorganisms during passage through the GIT. 2014 Published by Elsevier Masson SAS on behalf of Institut Pasteur.

A total of 234 LAB isolates from Brazilian food products were initially screened for their ability to survive at pH 2.0. Fifty one of the isolates survived and were selected. They were characterized by phenotypic methods, rep-PCR and identified using 16S rRNA gene sequencing as Lactobacillus fermentum (34 isolates), Lactobacillus plantarum (10) and Lactobacillus brevis (7). Based on being either highly tolerant to bile, showing an ability for auto-aggregation and/or hydrophobic properties, one L. fermentum (CH58), three L. plantarum (CH3, CH41 and SAU96) and two L. brevis (SAU105 and FFC199) were selected. The highest co-aggregation ability with Escherichia coli was observed to L. plantarum CH41. L. brevis SAU105 and FFC199 and L. fermentum CH58 exhibited antagonistic activity towards the pathogens Listeria monocytogenes and Staphylococcus aureus. L. plantarum CH3 and CH41 and L. brevis FFC199 showed adhesion ability to Caco-2 cells (1.6, 1.1 and 0.9%, respectively) similar to the commercial probiotic, Lactobacillus rhamnosus GG (1.5%). They were able to increase the transepithelial electrical resistance (TEER) of Caco-2 cells over 24 h (p

Yakupa is a traditional non-alcoholic cassava beverage produced by Brazilian Amerindians. In this work the microbial dynamics and metabolites involved in yakupa fermentation were investigated by PCR-denaturing gradient gel electrophoresis and chromatography analysis, respectively. The lactic acid bacteria (LAB) population was higher than yeast in the beginning of fermentation (5 log CFU mL(-1) and 3 log CFU mL(-1), respectively) and after 36 h both population increased reaching 7 log CFU mL(-1), remaining constant until 60 h. Culture dependent and independent methods in combination identified the bacteria Lactobacillus fermentum, L. plantarum, Weissela cibaria and W. confusa, and yeasts Saccharomyces cerevisiae and Pichia kudriavzevii. Maltose (41.2 g L(-1)), ethanol (6.5 g L(-1)) and lactic acid (7.8 g L(-1)) were the most abundant compounds identified by high performance liquid chromatography. Aldehydes, acids, alcohols and esters were identified by gas chromatography flame ionization detection. By the metabolites and PCA analysis we may assign the beverage's flavor to the microbial metabolism. Heterolactic LAB and S. cerevisiae dominated the yakupa fermentation, being responsible for the organoleptic characteristics of the final product. This is the first time that the microbial dynamics and physicochemical parameters were investigated in the yakupa beverage and it may contribute to the future selection of starter cultures to perform yakupa fermentations.

This study aimed to select autochthonous lactic acid bacteria (LAB) with probiotic and functional properties from goat dairies and test their addition to artisanal cheese for the inhibition of Salmonella typhi. In vitro tests, including survival in the gastrointestinal tract (GIT), auto- and co-aggregation, the hemolytic test, DNase activity, antimicrobial susceptibility, antibacterial activity, tolerance to NaCl and exopolysaccharide (EPS), gas and diacetyl production were conducted for sixty isolates. Based on these tests, four LAB isolates (UNIVASF CAP 16, 45, 84 and 279) were selected and identified. Additional tests, such as production of lactic and citric acids by UNIVASF CAP isolates were performed in addition to assays of bile salt hydrolase (BSH), β-galactosidase and decarboxylase activity. The four selected LAB produced high lactic acid (>17 g/L) and low citric acid (0.2 g/L) concentrations. All selected strains showed BSH and β-galactosidase activity and none showed decarboxylase activity. Three goat cheeses (1, 2 and control) were produced and evaluated for the inhibitory action of selected LAB against Salmonella typhi. The cheese inoculated with LAB (cheese 2) decreased 0.38 log10 CFU/g of S. Typhy population while in the cheese without LAB inoculation (cheese 1) the pathogen population increased by 0.29 log units. Further, the pH value increased linearly over time, by 0.004 units per day in cheese 1. In the cheese 2, the pH value decreased linearly over time, by 0.066 units per day. The cocktail containing selected Lactobacillus strains with potential probiotic and technological properties showed antibacterial activity against S. typhi in vitro and in artisanal goat cheese. Thus, goat milk is important source of potential probiotic LAB which may be used to inhibit the growth of Salmonella population in cheese goat, contributing to safety and functional value of the product.

Spontaneous cocoa bean fermentations performed under bench- and pilot-scale conditions were studied using an integrated microbiological approach with culture-dependent and culture-independent techniques, as well as analyses of target metabolites from both cocoa pulp and cotyledons. Both fermentation ecosystems reached equilibrium through a two-phase process, starting with the simultaneous growth of the yeasts (with Saccharomyces cerevisiae as the dominant species) and lactic acid bacteria (LAB) ( Lactobacillus fermentum and Lactobacillus plantarum were the dominant species), which were gradually replaced by the acetic acid bacteria (AAB) ( Acetobacter tropicalis was the dominant species). In both processes, a sequence of substrate consumption (sucrose, glucose, fructose, and citric acid) and metabolite production kinetics (ethanol, lactic acid, and acetic acid) similar to that of previous, larger-scale fermentation experiments was observed. The technological potential of yeast, LAB, and AAB isolates was evaluated using a polyphasic study that included the measurement of stress-tolerant growth and fermentation kinetic parameters in cocoa pulp media. Overall, strains L. fermentum UFLA CHBE8.12 (citric acid fermenting, lactic acid producing, and tolerant to heat, acid, lactic acid, and ethanol), S. cerevisiae UFLA CHYC7.04 (ethanol producing and tolerant to acid, heat, and ethanol), and Acetobacter tropicalis UFLA CHBE16.01 (ethanol and lactic acid oxidizing, acetic acid producing, and tolerant to acid, heat, acetic acid, and ethanol) were selected to form a cocktail starter culture that should lead to better-controlled and more-reliable cocoa bean fermentation processes.

Theobroma cacao seeds are the main raw material for chocolate production. During their fermentation, a succession of microorganisms are responsible for the physicochemical changes occurring in the pulp and inside the beans. The aim of this study was to investigate the effects of yeast inoculation (Saccharomyces cerevisiae UFLA CA11, Pichia kluivery CCMA0237, and Hanseniaspora uvarum CCMA0236) on the profile of the volatile compounds and microbial communities in cocoa fermentation. The resulting chocolate was also evaluated by temporal dominance of sensations (TDS) analyses. The dominant microorganisms during spontaneous fermentation were S. cerevisiae, H. uvarum, H. guilliermondii, Lactobacillus fermentum, Pediococcus sp., and Acetobacter pasteurianus. Similarly, S. cerevisiae, P. kluyveri, Candida sp., Pediococcus sp., and A. pasteurianus were the predominant microorganisms assessed by Denaturing Gradient Gel Electrophoresis (DGGE) in inoculated fermentation. Sixty-seven volatile compounds were detected and quantified by gas chromatography/mass spectrometry (GC/MS) at the end of fermentation and chocolates. The main group of volatile compound found after the inoculated and spontaneous fermentations was esters (41 and 39 %, respectively). In the chocolates, the main group was acids (73 and 44 % from the inoculated and spontaneous fermentations, respectively). The TDS analyses showed a dominance of bitter and cocoa attributes in both chocolates. However, in the inoculated chocolate, lingering fruity notes were more intense, while the chocolate produced by spontaneous fermentation was more astringent. Thus, the inoculation of yeast influenced the microbial profile, which likely affected the volatile compounds that affect sensory characteristics, resulting in chocolate with dominant bitter, cocoa, and fruity attributes.