Fermented Cocoa Research Articles

Effect of biologically upgraded cocoa pod husk meal on growth, serum and antioxidant properties of two rabbit breeds

BackgroundThe study evaluated the growth performance, haemabiochemical and antioxidant profile of two breeds of rabbits fed diets containing varying levels of Rhizopus stolonifer fermented cocoa pod husk meal (FCPHM). Cocoa pod husk meal was taken through solid-state fermentation for a period of 2 weeks using Rhizopus stolonifer as its starter culture. One hundred and eighty, 5-week-old unsexed weaned rabbits 90 New Zealand white (NZW) and 90 Chinchilla (CHL) were randomly allocated into three dietary treatments, Diets I, II and III (0%, 12.5 and 25%FCPHM, respectively), and were arranged in a 2 × 3 factorial design (2 breeds and 3 diets) of a randomized block design.ResultsThe study showed that the breed effect and dietary treatment were significant (p < 0.01) for total weight gain and feed conversion ratio with NZW breed fed 12.5% FCPHM having higher values than CHL breed. Serum total protein concentration was significantly (p < 0.05) influenced by the dietary treatment, while alanine aminotransferase, globulin and total protein concentrations were significantly (p < 0.01) affected by the breed effect. The interactive effect of breed and dietary treatment was only significant (p < 0.05) for serum albumin concentration. Malondialdehyde concentration was significantly influenced by both dietary treatments (p < 0.05) and breed (p < 0.001) factors.ConclusionIt was observed that CHL rabbits were more predisposed to stress than the NZW rabbits as indicated by the lipid peroxidation values, and according to this present finding, 12.5% FCPHM is recommended as active feed ingredient in rabbit diets.

Effects of Processing Time and Temperature on Flavanol and Procyanidin, Proanthocyanidin and Antioxidant Activity of Cocoa Bean in Taiwan

Flavanol in cocoa is beneficial to cardiovascular health. This study investigated the effect of roasting conditions, temperature ranging from 110 to 150°C and time duration 15-35 min, on the level of flavanol and procyanidin (FP), total proanthocyanidin (PAC) and antioxidant activity of fermented cocoa beans in Southern Taiwan. The content of FP in unroasted and roasted cocoa bean was determined by the normal phase HPLC method and the antioxidant activity was assayed by DPPH and ABTS methods. High roasting temperature for long time decreased the content of FP and PAC in cocoa beans, where ranged from 55.30±1.95 mg (-)-epicatechin equivalents /g and 103.56±8.33mg procyanidin B1/g (at 110°C for 15 min) to 19.12±0.66 mg (-)-epicatechin equivalents /g and 51.58±0.86 mg procyanidin B1/g (at 150°C for 35 min), respectively. Meanwhile, as the roasting temperature rose, the DPPH and ABTS free radical scavenging ability of roasted cocoa bean extracts decreased gradually, varying from the maximum (IC50 = 104±2 µg/ml and 50.4±1.1mg TE/g) for unroasted bean to the minimum (IC50 = 144±4 µg/ml and 27.7±2.4 mg TE/g), and then slightly increased to IC50=134±4 µg/ml and 31.5±0.7 mg TE/g, respectively at most severe roasting treatment, showing the formation of melanoidins and polyphenols caused the synergistic effect, leading to the rise of the antioxidant activity. The first order kinetic reaction and the Arrhenius equation were applied to forecast the change of FP concentration during roasting. The activation energy of FP degradation is 11.2 kcal/mol. The obtained kinetic equation could be useful to predict the remaining amount of FP in cocoa beans under the designed experiment. The cocoa products with higher flavanol content and antioxidant activity could be possible by setting appropriate roasting conditions.

Effect of cocoa bean origin and conching time on the physicochemical and microstructural properties of Indonesian dark chocolate

Abstract Indonesian cocoa is cheaper and considered second grade compared with most other cocoa. However, the domestic chocolate industry is not well-developed due to significantly low consumption. To cope with these issues, product innovation through technical process improvement is required to stimulate the domestic chocolate industry. This study aimed to investigate the effect of cocoa bean origin and conching time on the physicochemical (water content, texture, color, crude fat content, and melting enthalpy) and microstructural properties of chocolate. The experiment was conducted under a completely randomized factorial design consisting of two factors: cocoa bean origin (100% fermented cocoa beans from Jember, 100% fermented cocoa beans from Southeast Sulawesi, 50% fermented cocoa beans from Jember + 50% non-fermented cocoa beans from Southeast Sulawesi, and 50% fermented cocoa beans from Southeast Sulawesi + 50% non-fermented cocoa beans from Southeast Sulawesi) and conching time (4, 6 and 8 h). The results showed that cocoa bean origin significantly affected the hardness, gumminess and color of chocolate, including the redness and yellowness level, whereas conching time affected water content, hardness, cohesiveness, elasticity, and crude fat content. Differential scanning calorimetry (DSC) analysis showed that the treatment with 100% fermented cocoa beans from Jember presented higher values of Tonset, ΔHmelt and area compared with those of the treatment with 100% fermented cocoa beans from Southeast Sulawesi, which presented higher values of Tpeak and Tend. Scanning Electron Microscopy (SEM) analysis showed a distribution of solid and dense particles with crystal interaction with chocolate structures.

Use of the &amp;lt;sup&amp;gt;13&amp;lt;/sup&amp;gt;C/&amp;lt;sup&amp;gt;12&amp;lt;/sup&amp;gt;C, &amp;lt;sup&amp;gt;15&amp;lt;/sup&amp;gt;N/&amp;lt;sup&amp;gt;14&amp;lt;/sup&amp;gt;N and &amp;lt;sup&amp;gt;18&amp;lt;/sup&amp;gt;O/&amp;lt;sup&amp;gt;16&amp;lt;/sup&amp;gt;O Isotopic Ratios of Theobromine and Caffeine in the Characterization of Geographic Origin

The aim of this work is to characterize the geographical origin of cocoa beans and coffee beans. This study aims to contribute to the traceability of raw materials in order to fight against falsification. For this purpose, we based our work on the measurement of isotope ratios in 13C, 15N and 18O. The multi-element isotope ratios have been evaluated as a means to distinguish fermented cocoa beans of different geographic and varietal origins. The isotopic ratios of 13C, 15N and 18O were measured in theobromine obtained from samples of fermented cocoa beans. Twenty-two (22) samples of different geographical origins covering the four mainland cocoa producing areas were analyzed on the one hand and on the other hand, 16 caffeine samples from various origins were also analyzed. The treatment of the values resulting from these isotopic analyzes by statistical methods, namely the principal component analysis (PCA) makes it possible to visualize the discriminations between the different origins. The most discriminating variables identified as responsible for the geographic and varietal differences were the δ15N, δ13C and δ18O values of cocoa beans and certain extracts and tissues. We have shown that the isotope ratios are correlated with the altitude and precipitation conditions encountered in the different cocoa growing regions.

Isolation and identification of microbial species found in cocoa fermentation as microbial starter culture candidates for cocoa bean fermentation in Colombia.

Microbial activity involved in the cocoa beans fermentation process is essential to maintain and improve the organoleptic and nutritional qualities of chocolate; therefore, the aim of this investigation was to search and select microbial isolates with the potential to improve the quality of cocoa beans. Fermentation experimentswere conducted on farms located in Maceo (Antioquia), San Vicente de Chucurí (Santander), and Rivera and Algeciras (Huila), Colombia. Yeast, lactic acid bacteria (LAB), acetic acid bacteria (AAB), and mesophilic aerobic microorganisms were obtained from different fermentation batches. The growth of these microorganismswas tested in six treatments as follows: 50% cocoa pulp agar (CPA), high concentrations of glucose (10%), ethanol (5%), and acetic acid (7%), an acidic pH of 3.0, and a high temperature of 50oC for 24 h. The isolates with the highest growth were identified by 18S and 16S rRNA gene analysis, revealing a high diversity ofspecies associated with cocoa fermentation, including eight species of yeasts (Debaryomyces hansenii, Meyerozyma guillermondii, Wickerhanomyces anomalus, Pichia guillermondii, Pichia kudriavzevii, Trichosporon asahii, Candida parapsilosis, and Pichia manshurica), six species of LAB (Pediococcus acidilactici, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus farraginis, Lactobacillus rhamnosus, and Leuconostoc mesenteroides), four species of AAB (Gluconobacter japonicus, Acetobacter tropicalis, Acetobacter pasteurianus, and Acetobacter malorum/tropicalis), and three species of Bacillus spp. (Bacillusaryabhattai /megaterium, Bacillus subtilis, and Bacillus coagulans). In general, microbial populations increased in cocoa batches after 12 h of fermentation and decreased after 84-96 h. All the yeast isolates grew in 10% glucose and CPA, 85.7% in 5% ethanol, and 95% at a pH of 3.0. All the yeast isolates were affectedby 7% acetic acid and incubation at 50oC for 24 h. Eighty-five percent of the LAB grew in 10% glucose, 100% in 5% ethanol, 42.8% in CPA, 64% at a pH of 3.0, and 35.7% grew after being exposed to 50oC for 24 h; all were affected by 7% acetic acid. As for the AAB, 100% grew in 10% glucose, 71% in 7% ethanol, 100% grew in CPA, in 7% acetic acid, and at a pH of 3.0, while 100% were affected by incubation at 50oC. Three yeast isolates, W. anomalus, D. hansenii and M. guillermondii, three LAB isolates, P. acidilactici, L. brevis, and L. plantarum, and three AAB isolates, A. tropicalis, A. pasteurianus and G. japonicus, were selected as promising strains to be used in a microbial starter culture for cocoa bean fermentation to improve the organoleptic quality of cocoa.

Understanding amino acids and bioactive amines changes during on-farm cocoa fermentation

Fermentation is of utmost relevance in cocoa processing. Limited information is available on the impact of fermentation on nitrogenous compounds which are associated with relevant health benefits and aroma quality of chocolate. The objective of this study was to investigate the changes on free amino acids and bioactive amines during cocoa on-farm fermentation. In unfermented cocoa 14 amino acids and five amines were detected, with predominance of glutamic acid, leucine, phenylalanine, serine and spermidine and serotonin, respectively. Fermentation followed its due course, confirmed by total titratable acidity, pH and temperature changes. Amino acids levels varied during fermentation and the final levels were higher compared to the unfermented cocoa. Hydrophobic amino acids were prevalent at the end of fermentation. Total levels of most amines decreased (serotonin disappeared) during fermentation; whereas putrescine remained constant and phenylethylamine increased. Multivariate analysis differentiated three distinct phases for these nitrogenous compounds during fermentation: (i) from 0 to 48 h – high serotonin, tryptamine, and pH; (ii) from 60 to 132 h – high phenylalanine, lysine, leucine, alanine, threonine, phenylethylamine, and acidity; and (iii) 144 h – high glycine, histidine, arginine and phenylethylamine. The possibility of using blends with cocoa from different fermentation times, can lead to fermented cocoa with unique functional and sensory properties.

Exploring the Capability of Yeasts Isolated from Colombian Fermented Cocoa Beans to Form and Degrade Biogenic Amines in a Lab-Scale Model System for Cocoa Fermentation

Yeast starters for cocoa fermentation are usually tested according to their enzymatic activities in terms of mucilage degradation and flavor improvement, disregarding their influence on the production or elimination of toxic compounds as biogenic amines (BAs), important for human health. In this work, we tested 145 strains belonging to 12 different yeast species and isolated from the Colombian fermented cocoa beans (CB) for their capability of producing BAs in vitro. Sixty-five strains were able to decarboxylate at least one of the amino acids tested. Pichia kudriavzevii ECA33 (Pk) and Saccharomyces cerevisiae 4 (Sc) were selected to evaluate their potential to modulate BAs, organic acids, and volatile organic compounds (VOCs) accumulation during a simulated cocoa fermentation. The growth of Sc or Pk in the presence of CB caused a significant reduction (p < 0.05) of 2-phenylethylamine (84% and 37%) and cadaverine (58% and 51%), and a significant increase of tryptamine and putrescine with a strong influence of temperature in BA formation and degradation. In addition, our findings pointed out that Pk induced a major production of fatty acid- and amino acid-derived VOCs, while Sc induced more VOCs derived from fatty acids metabolism. Our results suggest the importance of considering BA production in the choice of yeast starters for cocoa fermentation.

Classification of fermented cocoa beans (cut test) using computer vision

Fermentation of cocoa beans is a critical step for chocolate manufacturing, since fermentation influences the development of flavour, affecting components such as free amino acids, peptides and sugars. The degree of fermentation is determined by visual inspection of changes in the internal colour and texture of beans, through the cut-test. Although considered standard for evaluation of fermentation in cocoa beans, this method is time consuming and relies on specialized personnel. Therefore, this study aims to classify fermented cocoa beans using computer vision as a fast and accurate method. Imaging and image analysis provides hand-crafted features computed from the beans, that were used as predictors in random decision forests to classify the samples. A total of 1800 beans were classified into four grades of fermentation. Concerning all image features, 0.93 of accuracy was obtained for validation of unbalanced dataset, with precision of 0.85, recall of 0.81. Although the unbalanced dataset represents actual variation of fermentation, the method was tested for a balanced dataset, to investigate the influence of a smaller number of samples per class, obtaining 0.92, 0.92 and 0.90 for accuracy, precision and recall, respectively. The technique can evolve into an industrial application with a proper integration framework, substituting the traditional method to classify fermented cocoa beans.

Co-culturing fructophilic lactic acid bacteria and yeast enhanced sugar metabolism and aroma formation during cocoa beans fermentation

Glucose and fructose are the main fermentable sugars in cocoa pulp. During fermentation, glucose is consumed within 48–72 h and fructose only after 120 h, mainly associated with the preferential use of glucose by microorganisms. In the first stage of this study, the complete genome sequence of a lactic acid bacterium with high fructose consumption capacity (Lactobacillus plantarum LPBF35) was reported. The notable genomic features of L. plantarum LPBF35 were the presence of alcohol/acetaldehyde dehydrogenase gene and improved PTS system, confirming its classification as a “facultatively” fructophilic bacterium. Subsequently, this bacterium was introduced into cocoa fermentation process in single and mixed cultures with Pediococcus acidilactici LPBF66 or Pichia fermentans YC5.2. Community composition by Illumina-based amplicon sequencing and viable counts indicated suppression of wild microflora in all treatments. At the beginning of the fermentation processes, cocoa pulp consisted of approximately 73.09 mg/g glucose and 73.64 mg/g fructose. The L. plantarum LPBF35 + P. fermentans YC5.2 process showed the lowest levels of residual sugars after 72 h of fermentation (7.89 and 4.23 mg/g, for fructose and glucose, respectively), followed by L. plantarum LPBF35 + Ped. acidilactici LPBF66 (8.85 and 6.42 mg/g, for fructose and glucose, respectively), single L. plantarum LPBF35 treatment (4.15 and 10.15 mg/g, for fructose and glucose, respectively), and spontaneous process (22.25 and 14.60 mg/g, for fructose and glucose, respectively). The positive interaction between L. plantarum LPBF35 and P. fermentans YC5.2 resulted in an improved formation of primary (ethanol, lactic acid, and acetic acid) and secondary (2-methyl-1-butanol, isoamyl acetate, and ethyl acetate) metabolites during fermentation. The primary metabolites accumulated significantly in cocoa beans fermented by P. fermentans YC5.2 + L. plantarum LPBF35, causing important reactions of color development and key flavor molecules formation. The results of this study suggest that fructophilic lactic acid bacteria and yeast is a microbial consortium that could improve sugar metabolism and aroma formation during cocoa beans fermentation.

Packaging and storage of cocoa beans fermented with Lactobacillus plantarum HL-15 in Agricultural Technology Park Nglanggeran, Yogyakarta

In Gunungkidul, there is Agricultural Techno Park (ATP) Nglanggeran which processes fermented cocoa beans from local farmers for making chocolate products. This research aimed to obtain a suitable packaging method for fermented cocoa beans during storage in ATP Nglanggeran. This research was carried out using a completely randomized factorial design. The first factor was the addition of starter (B), which consisted of B1 (with Lactobacillus plantarum HL 15) and B2 (without Lactobacillus plantarum HL 15). The second factor was the packaging methods (K), which consisted of K1 (vacuumed PP plastic 0.8 mm in thickness), K2 (non-vacuumed PP plastic 0.8 mm in thickness), K3 (plastic container), K4 (vacuumed PP plastic 0.8 mm in thickness stored within the plastic container), and K5 (nylon sack). The third factor was the storage time (P), which were P0 (0 month), P1 (1 month), and P2 (2 months). The result showed that the samples with the addition of the starter had a lower number of fungal colonies than those without the addition. Besides, the packaging using vacuumed PP plastic 0.8 mm in thickness stored within a plastic container gave the best result on minimizing the total fungal growth contamination and the peroxide value.

Pod storage time and spontaneous fermentation treatments and their impact on the generation of cocoa flavour precursor compounds

SummaryMost of volatile flavour compounds occurred during the cocoa postharvest operations including pod storage and fermentation whom effects on these metabolites were aimed by the present study. Three different spontaneous fermentation treatments were performed on the farm, and the main isolated yeasts were identified using PCR‐DGGE technique. Fermented cocoa beans were solar‐dried, and their flavour compounds were identified using solid phase microextraction technique–gas chromatography–mass spectrometry. The main results showed that Pichia kudriavzevii was common yeast involved in the cocoa fermentation. Thirty flavour precursor compounds grouped into six chemical families including alcohols and pyrazines were identified in raw cocoa samples. ACP showed that short durations of pod storage and fermentation promote the formation of flavour precursors of cocoa. No spontaneous cocoa fermentation treatment influenced the occurrence of aroma compounds. The generation of flavour compounds depends only on the pod storage and the fermentative microorganisms of cocoa.

The Contribution of Fermentation Technology to the Value Added of Certification Cocoa in the Subak Abian Amerta Nadi Tuwed Village Melaya District Jembrana Regency

The development of cocoa as an export commodity has become an important issue in the global trade sector, where cocoa is one of the export commodities. The implementation of a sustainable certification program with fermentation processing is very important, this is because consumers' demands are increasingly critical, especially in the aspects of quality, health and product safety to how the product can be socially beneficial and not cause negative impacts on the environment.&#x0D; The study was conducted on sustainable certification cocoa farmers in Jembrana Regency. Determination of respondents carried out intentionally (purposive sampling) as many as 40 respondents did cocoa fermentation and as many as 30 respondents who did not ferment cocoa taken by survey methods. Data analysis techniques using different test analysis and value added analysis using the Hayami method. &#x0D; The results showed that the income of cocoa farmers experienced a significant increase in the fermentation and non-fermentation processing. The fermentation production process with an output of 118 kg gets a profit of Rp. 8.911,- with a percentage or level of profit of 39,27 percent of the output value of Rp. 20.911, so from the calculation of profit margins in the cocoa fermentation process the net profit received by farmers is Rp. 8.065,- per production process. Based on the analysis of farmer's income and analysis of added value of cocoa fermentation shows that there is an increase in farmer's income and there is added value obtained by farmers who carry out cocoa fermentation, it provides benefits for cocoa farmers in Jembrana District in addition to increasing income, cocoa added value, and can maintain quality cocoa. It is expected that farmers who participate in certification will conduct fermentation processing to maintain the quality of cocoa beans and increase income, it is necessary to support from local and provincial governments as well as other parties as a steakholder providing technical guidance to cocoa farmers.

Development of a model for the alcoholic fermentation of cocoa beans by a Saccharomyces cerevisiae strain

The aromatic quality of chocolate requires the use of cocoa with high aromatic potential, this being acquired during the fermentation of cocoa beans. Traditional fermentation is still often carried out on a small scale with wild strains of yeasts and acetic bacteria and under poorly controlled conditions leading to cocoa quality ranging from best to worst. This study is the first part of a project aiming to control quality of cocoa to produce high aromatic quality chocolate by using a mixed starter of selected strains of yeast and acetic bacteria and by controlling the conditions of fermentation. To achieve this objective, a mathematical model of the alcoholic fermentation of cocoa beans has been developed. The growth, glucose consumption and ethanol production of Saccharomyces cerevisiae LM strain in synthetic broth were modeled for the most important intrinsic (pH, glucose, ethanol, free nitrogen and oxygen levels) and extrinsic (temperature, oxygen level) fermentation parameters. The model was developed by combining the effects of individual conditions in a multiplicative way using the gamma concept. The model was validated in liquid synthetic medium at two different inoculation levels 104 and 106 CFU/mL with an increase in temperature that recorded during spontaneous fermentations. The model clearly shows that the level of inoculation and the speed of the increase in temperature clearly drive yeast growth, while other factors including pH and ethanol, free nitrogen and oxygen levels have no significant impact on yeast growth.

Potential of Bacteria from Alternative Fermented Foods as Starter Cultures for the Production of Wheat Sourdoughs.

Microbial strains for starter culture-initiated sourdough productions are commonly isolated from a fermenting flour–water mixture. Yet, starter culture strains isolated from matrices other than sourdoughs could provide the dough with interesting metabolic properties and hence change the organoleptic properties of the concomitant breads. Furthermore, the selection of sourdough starter cultures does not need to be limited to lactic acid bacteria (LAB), as other food-grade microorganisms are sometimes found in sourdoughs. Therefore, different strains belonging to LAB, acetic acid bacteria (AAB), and coagulase-negative staphylococci (CNS) that originated from different fermented food matrices (fermenting cocoa pulp-bean mass, fermented sausage, and water kefir), were examined as to their prevalence in a wheat sourdough ecosystem during 72-h fermentations. Limosilactobacillus fermentum IMDO 222 (fermented cocoa pulp-bean mass isolate) and Latilactobacillus sakei CTC 494 (fermented sausage isolate) seemed to be promising candidates as sourdough starter culture strains, as were the AAB strains Acetobacter pasteurianus IMDO 386B and Gluconobacter oxydans IMDO A845 (both isolated from fermented cocoa pulp-bean mass), due to their competitiveness in the wheat flour-water mixtures. Wheat breads made with G. oxydans IMDO A845 sourdoughs were significantly darker than reference wheat breads.

Designing an Arduino-based Automatic Cocoa Fermentation Tool

Automatic cocoa fermentation design is expected to facilitate the work of cocoa farmers during the process of reversing and stirring cocoa fermentation based on the right temperature. Fermentation process is of course done in a box or sack so that chocolate quickly produces heat and is cemented. However, in certain conditions, especially when in sacks there are often obstacles in the stirring process. Often the fermented chocolate experiences weathering or moldyness due to the uneven reversal that causes chocolate to clot, causing weathering or moldiness and produce an unpleasant odor and unattractive color on the cocoa beans. To overcome this problem a tool that automatically can turn or stir the cocoa beans evenly. This device is controlled by Arduino Uno R3 with a sensor that is an LM35 temperature sensor and has an LCD output and DC motor. This tool uses Relay to adjust the delay when driving a DC motor. The working principle of this tool, when the LM35 temperature sensor receives heat conditions on the cocoa beans, the LCD will display the condition of the temperature while the relay will instruct the DC motor to move the Cocoa Fermentation rail rotating left or right. The purpose of making this tool is to create a tool that can help alleviate the work of cocoa farmers in cocoa bean stirring activities at the time of cocoa bean fermentation controlled by Arduino.

Profile of bioactive compounds, antioxidant and aromatic component from several clones of cocoa beans during fermentation

Commonly cocoa beans containing various chemical components, nutrients, and bioactive compound. The main bioactive compounds in cocoa beans consisting flavonoids and methylxantines has an positive influence on the health of the human body. Besides the fermented cocoa beans also produce a distinctive of cocoa aromatic. One of the main factors influencing the formation of aroma, chemical content and bioactive compounds of cocoa beans is the fermentation process. The purpose of research is to determine the effect of fermentation duration of cocoa clones with different varieties on the profile of bioactive compounds, antioxidant activity and aromatic components (volatiles) that occur in cocoa beans during fermentation. The treatment was arranged in a Randomized Block Design of two factors, the first factor is cocoa clones (Sulawesi 2/S2 clones and local clones) and the second factor is fermentation time (0, 2, 4 and 6 days). The observed parameters were polyphenol content, antioxidant activity, flavonoid bioactive compounds (catechin and epicatechin) and methylxanthine (theobromin and caffeine), as well as aromatic components (volatile) of fermented cocoa beans. The results of the study was showed that cocoa beans from local clones providing polyphenols, antioxidant activity and bioactive compounds (catechin, epicatechin and theobromine) are higher than S2 clones. While the high caffeine content and distinctive aroma of the pyrazine compound were produced from S2 cocoa beans on 6 days fermentation. The average content of polyphenols, antioxidants and bioactive compounds in non-fermented cocoa beans is higher than fermented beans, but the resulting scent is not strong. The results of the research are expected to obtain information about the profile of bioactive compounds, antioxidants and aromatic components of cocoa beans especially derived from Pinrang district has the potential improving community health.

NMR-based Lavado cocoa chemical characterization and comparison with fermented cocoa varieties: Insights on cocoa’s anti-amyloidogenic activity

The metabolic profile of Lavado cocoa was characterized for the first time by NMR spectroscopy, then compared with the profiles of fermented and processed varieties, Natural and commercial cocoa.The significant difference in the contents of theobromine and flavanols prompted us to examine the cocoa varieties to seek correlations between these metabolite concentrations and the anti-amyloidogenic activity reported for cocoa in the literature.We combined NMR spectroscopy, preparative reversed-phase (RP) chromatography, atomic force microscopy, in vitro biochemical and cell assays, to investigate and compare the anti-amyloidogenic properties of extracts and fractions enriched in different metabolite classes.Lavado variety was the most active and the catechins and theobromine were the chemical components of cocoa hindering Aβ peptide on-pathway aggregation and toxicity in a human neuroblastoma SH-SY5Y cell line.

Characterization of Odorants Causing Smoky Off-Flavors in Cocoa.

Application of an aroma extract dilution analysis in parallel applied to the volatiles isolated from a sample of fermented cocoa with a pronounced smoky off-flavor and a reference sample with flawless aroma revealed seven potential off-flavor compounds with smoky/phenolic odor qualities and higher flavor dilution factors in the off-flavor sample than in the reference sample. These compounds were 2-methoxyphenol, 3- and 4-methylphenol, 3- and 4-ethylphenol, and 3- and 4-propylphenol. Their quantitation in seven off-flavor samples and two reference samples without off-flavor showed that 4-propylphenol did not exceed its odor threshold value. From the concentrations obtained for the other six compounds and their odor threshold values in deodorized cocoa butter, maximum tolerable concentrations were derived for the incoming goods inspection in the chocolate industry. The suggested limits are 2 μg/kg for 3-ethylphenol and 3-propylphenol, 20 μg/kg for 4-methylphenol, 3-methylphenol, and 4-ethylphenol, and 70 μg/kg for 2-methoxyphenol.

Aislamiento de microorganismos autóctonos para formular un inoculo definido para la fermentación del cacao en pequeña escala

In cocoa fermentation exists a wide microbial diversity; the most important microorganisms are yeasts, lactic acid bacteria (LAB), and acetic acid bacteria (AAB). Such diversity can result that fermented cocoa quality is not always the same. The use of microbial consortia can steer the process, allowing the production of fermented cocoa beans with homogeneous quality and safety. For that reason, it was proposed to use indigenous microorganisms to formulate defined inoculums to conduct the small- fermentation of Mexican cocoa. A total of 54 strains were isolated from the spontaneous fermentation of cocoa. These included yeasts (Candida, Rhodotorula, Saccharomyces and Yarrowia), LAB (Lactobacillus and Lactococcus), and AAB (Acetobacter and Gluconobacter). The cocoa fermentations were inoculated with an inoculum composed of lipolytic Yarrowia, Lactococcus lactis, and Acetobacter aceti, only varying the form of inoculation (mixed or microbial succession) and were compared with spontaneous fermentation. It was observed that fermentation conducted by succession inoculation form showed similar behavior to the spontaneous process, obtaining well-fermented cocoa beans with homogeneous quality and safety. &nbsp

Effect of Addition of a Specific Mixture of Yeast, Lactic and Acetic Bacteria in the Fermentation Process to Improve the Quality and Flavor of Cocoa Beans in Colombia

Cocoa fermentation process is fundamental to generate flavors and aromas that are characteristics of chocolate. In Colombia, this process is carried out by microbiota that spontaneously colonizes cocoa beans, therefore the quality of the fermentation is inconsistent. Taking into account that the fermentation of cocoa beans is carried out by a consortium of microorganisms, the aim of thisresearch was to describe the effect of the addition of a specific mixture of yeasts, acetic acid bacteria, and lactic acid bacteria on the physicochemical and organolepticcharacteristics of cocoa beans (clone CCN 51). Isolates of two yeasts (Wickerhamomyces anomalus and Debaryomyces hansenii), three acetic acid bacteria (AAB), (Gluconobacter japonicus, Acetobacter tropicalis, and Acetobacter pasteurianus) and three lactic acid bacteria (LAB) (Pediococcus acidilactici, Lactobacillus brevis, and Lactobacillus plantarum) obtained fromprevious cocoa fermentations selected for their pectinases and acid production capacities in a specific mixture were used. Using the micro-fermentation technique, the effect of a biological starter was evaluated under different viable microorganismsratios (Yeasts: LAB: AAB as follows, 1: 1: 1, 1: 2: 2, 1: 2: 1, 1: 1: 2, 2: 1: 1, 2: 2: 1, 2: 1: 2, and 2: 2: 2). The concentration of each microorganism was standardized at 1x107 cfu/mL, then the biomass of 4 mL for ratio 1 and 8 mL for ratio 2 of each suspension of microorganisms was added at time zero. Different doses of inoculum were 0%, 1%, 2%, 3%, 4%, and 5% v/w mL inoculum/g cocoa beans. A beneficial effecton the sensory quality of cocoa beans was evidenced by the addition of microorganisms; the best proportion of microorganisms was 2:1:2 (yeasts:LAB:AAB) and the best inoculum dose was 3% (v/w) showing lower acidity, astringency, and bitterness, and emphasizing the cocoa flavors, fruity, nutty, and panela malt.