Fermented Cocoa Research Articles

Dynamics of volatile compounds in TSH 565 cocoa clone fermentation and their role on chocolate flavor in Southeast Brazil.

The effects of indigenous fermentation on volatile compound profiles in a Theobroma cacao L, TSH565 clone, resistant to Moniliophtora perniciosa and Phytophthora spp. were evaluated in Southern Brazil. Sixty-three volatile flavor compounds in pulp and 36 in grains were identified by SPME-HS/GC-MS and classified as terpenes, alcohols, esters, ketones and aldehydes, among others. The relative amount of these compounds and their evolution until the end of the fermentation process were assessed in both fresh and fermented grains/pulp masses. β-myrcene and β-cis-ocimene, among terpenes, were detected in high amounts and are associated to a fine chocolate aroma. The sensory evaluation of chocolates manufactured from the fermented cocoa was performed by trained panelists, which defined 15 sensory descriptors. Chocolates from the TSH565 cultivar were characterized by a rich, fruity, intense cocoa flavor and bitterness, which are valuable sensorial and commercial attributes.

Chemometric approaches for postharvest quality tracing of cocoa: An efficient method to distinguish plant material origin

The aim of this study was to compare the quality of a mixture of cocoa harvested and fermented in three subregions of Antioquia (Colombia), from the chemometric profile based on multivariate statistical analysis. A mixture of clones CCN-52, ICS-1, FLE-2, and FEC-2 harvested in Bajo Cauca, Uraba and Magdalena Medio were subjected to a spontaneous fermentation. The characterization of raw and well-fermented cocoa was performed through 38 parameters, and results were compared by a Principal Component Analysis (PCA) and a Cluster Analysis (CA), followed by a Principal Factors Analysis (PFA- CA). The CA showed that there are differences among subregions only in raw cocoa from Bajo Cauca. PCA allowed identifying the variability between raw and fermented cocoa in a representative way and these results were consistent with the chemical profile. Besides, the number of parameters to differentiate raw cocoa from different subregions was reduced (11–13 parameters) and it was possible to characterize well fermented cocoa with only 10 parameters of 38. PFA-CA consolidated in three factors a grouping to identify the cocoa quality according to the process or interest of the sensory or functional properties. Factor 1 (cocoa quality indicators with functional properties), Factor 2 (indicators of quality of the beginning of fermentation) and Factor 3 (indicators of quality of well-fermented cocoa) each one with a weight of 39, 35 and 26 respectively.

Analytical performance for determination of synthetic pyrethroids residue in Indonesian fermented cocoa using GC-ECD

Pyrethroid insecticides are causing problems for the environment because they are toxic at such small concentrations and very difficult to determine. Characteristics of pyrethroids insecticides include high efficiency, wide spectrum, low biodegradability, and low mammalian and avian toxicity. They also demonstrate strong selectivity for insects and invertebrates. This study presents the results of the analysis conducted on four synthetic pyrethroids in fermented cocoa samples from Indonesia. The pesticides were extracted using acetonitrile and the analysis was carried out by Gas Chromatography with Electron Capture Detector (GC-ECD). Column programming temperature was conducted to get a complete separation. The total operation took less than 10 minutes to separate four standards of synthetic pyrethroids. The synthetic pyrethroids were identified by their retention times and quantified using an external calibration method. The linearity test was carried out for four standard synthetic pyrethroids, namely: lambda-cyhalothrin, permethrin, cypermethrin, and fenvalerate, from 1 μg/kg to 10 μg/kg. The coefficient of correlation (r) was found to be 0.9999; 0.9998; 0.9944; and 0.9999 for lambda-cyhalothrin, permethrin, cypermethrin, and fenvalerate, respectively. The detection limit for synthetic pyrethroids residue was found to be below the EU and Japan regulations, and the method can be applied for the determination of synthetic pyrethroids residues in fermented cocoa.

Effects of cocoa clones and fermentation times on physical and chemical characteristics of cocoa beans (Theobroma cacao L.)

The effects of cocoa clones and fermentation time on the physical and chemical characteristics of cocoa beans has been examined. This study used cocoa pods of superior clones, namely Sulawesi 1, Sulawesi 2, and MCC 02 clones cultivated in East Luwu, South Sulawesi, Indonesia. Prior to the fermentation, the mature cocoa pods were stored for six days. Fermentation took place for 24, 48, 72, 96, and 120 hours using fermentation boxes made of styrofoam with the capacity of box is 7 kg of wet beans. The drying of the cocoa beans used sun drying method. The results showed that the interaction between fermentation time and type of clones have significantly affected moisture content, proportion of cocoa nibs and shells, and fat content; but the interaction of the both had no significant effect on pH of cocoa nibs and FFA content. In general, the Sulawesi 2 clone had better physical and chemical characteristics than those of the Sulawesi 1 and MCC 02 clones.

Acceleration of cocoa fermentation through the action of bacteria (Acetobacter aceti) and yeast (Saccharomyces cerevisiae)

Introductionthe management of the curriculum in teacher training within the Catholic University of Salta in the School of Education and Teacher Improvement, this thought from a deductive perspective in which theoretical processes are established that allow the explanation of the curriculum in terms of teacher training and its impact on the organizational articulation. Objectiveto specify a strategic articulated intervention device that requires the constitution of a new emancipatory discourse. Materials and methodsWe present a qualitative research that analyzes the problems and variables established in the study. ResultsThe process of rupture leads us to reflect on the teaching position in the face of praxis, on the processes of "learning to teach" which involves continuous training. DiscussionThis perspective points to the development of certain skills that are developed over time of the formation through diverse strategies Conclusionsthe development of the professional competences of the teacher that confront with the disciplinary approach from an innovative glance

Analysis of Environmental Conditions Affecting Acids Production in Lactic Acid Bacteria Involved in Ivorian Cocoa Fermentation

Original Research Article Acetic and lactic acids produced during cocoa fermentation impact strongly the quality of fermented cocoa beans and chocolate. In this study, acid production from various carbon source and environmental conditions was analyzed in five lactic acid bacteria previously characterized as citrate lyase producers. It appears that production of acetic and lactic acids by these strains is strongly dependent on environmental conditions such as pH and temperature. Indeed, maximum yield of acids occurred at 30°C and pH 4 in Lactobacillus plantarum strain T11G3 and Lactobacillus casei strain T10G5. On the other hand acid production was maximum at 40°C and pH 4 in Lactobacillus plantarum strains T8N10, T10AG26 and at 40°C and pH 4.5 in Leuconostoc mesenteroides strain T8AB6. These strains also presented variable acid production regarding the sugar tested with sucrose allowing the highest acid yield (38.12 mg/mL) comparatively to that obtained from glucose (23.91 mg/mL) and fructose (26.28 mg/mL). Moreover, maximum acid production in all the strains was rapidly achieved, after 36-48h fermentation with sucrose, whereas a delayed production (around 72 h) was observed with glucose or fructose. Alcoholic conditions were found to particularly enhance acid production in the strains studied (Leuconostoc mesenteroides strain T8AB6). The maximum yield of acetic and lactic acids was 12.7 mg/mL and 14.4 mg/mL respectively in Leuconostoc mesenteroides strain T8AB6 and Lactobacillus plantarum strain T8N10 under citric acid condition. Overall, this study highlights that carbon source and environnemental condition impact on the production of acidic compounds by LAB from fermenting cocoa. Furthermore, variability in acid production was observed between the studied strains in most of the considered conditions. Thus, the different LAB strains studied probably behave as complementary consortium to continuously achieve the process of natural cocoa fermentation.

Simulated Gastrointestinal Digestion of Cocoa: Detection of Resistant Peptides and In Silico/In Vitro Prediction of Their Ace Inhibitory Activity.

In this study we investigated the oligopeptide pattern in fermented cocoa beans and derived products after simulated gastrointestinal digestion. Peptides in digested cocoa samples were identified based on the mass fragmentation and on the software analysis of vicilin and 21 KDa cocoa seed protein sequences, the most abundant cocoa proteins. Quantification was carried out by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) using an internal standard. Sixty five peptides were identified in the digested samples, including three pyroglutamyl derivatives. The in vitro angiotensin-converting enzyme (ACE)-inhibitory activity of cocoa digests were tested, demonstrating a high inhibition activity, especially for digestates of cocoa beans. The peptides identified were screened for their potential ACE inhibitory activity through an in silico approach, and about 20 di-, three- and tetra-peptides actually present in our samples were predicted as active. Two of the potentially active peptides were chemically synthesized and then assessed for their inhibitory activity by using the ACE in vitro assay. These peptides demonstrated an ACE inhibitory activity, however, that was too weak to explain alone the high activity of cocoa digestates, suggesting a synergic effect of all cocoa peptides. As a whole, results showed that an average chocolate portion (30 g) ensures an amount of peptides after digestion that, assuming complete absorption, could reach almost a complete inhibition of ACE.

Traceability of Functional Volatile Compounds Generated on Inoculated Cocoa Fermentation and Its Potential Health Benefits.

Microbial communities are responsible for the unique functional properties of chocolate. During microbial growth, several antimicrobial and antioxidant metabolites are produced and can influence human wellbeing. In the last decades, the use of starter cultures in cocoa fermentation has been pushed to improve nutritional value, quality, and the overall product safety. However, it must be noted that unpredictable changes in cocoa flavor have been reported between the different strains from the same species used as a starter, causing a loss of desirable notes and flavors. Thus, the importance of an accurate selection of the starter cultures based on the biogenic effect to complement and optimize chocolate quality has become a major interest for the chocolate industry. This paper aimed to review the microbial communities identified from spontaneous cocoa fermentations and focused on the yeast starter strains used in cocoa beans and their sensorial and flavor profile. The potential compounds that could have health-promoting benefits like limonene, benzaldehyde, 2-phenylethanol, 2-methylbutanal, phenylacetaldehyde, and 2-phenylethyl acetate were also evaluated as their presence remained constant after roasting. Further research is needed to highlight the future perspectives of microbial volatile compounds as biomarkers to warrant food quality and safety.

Analysis of the performance of a newly designed fermenter built in local materials for improvement of cocoa fermentation, in Ivory Coast

Objective: Cocoa beans fermentation is a spontaneous chemical process, traditionally done in box or in a heap. Equipment is also important in achieving the final chocolate aroma. This study analyzes the performances of a new-designed cocoa bean fermenter. Methodology and results: In this study, a new type of fermenter, a rotating cylindrical fermenter (RCF) has been designed in order to improve the fermentation of cocoa that remains difficult to control, because of the spontaneous nature of the microbiota. The performances of this fermenter was analyzed and compared to those of the traditional wooden box (TWB) fermenter that is commonly used on farm. During the 6 days of fermentation in both fermenters, the growth of microorganisms such as yeasts, lactic bacteria, acetic bacteria, bacillus and moulds as well as chemical and physical changes of the fermenting cocoa were monitored. The results showed that in the fermenter (RCF) a rapid temperature increase was observed in the course of the fermentation process with a temperature reaching 51°C within 73 h comparatively to the traditional fermenter (48°C within 118 h). This leads to a higher proportion of brown beans, indicator of a good fermentation from RCF fermenter as assessed by the cut test. This proportion was 94.44% for RCF and 85.88% for TWB. Moreover the optimization of heat generated in the RCF fermenter, allowed a normal browning (final gray level was 77 in both fermenters) despite modification of microbiota growth order (early growth of acetic bacteria and stunted growth of yeast in RCF, but not for TWB). Conclusion and application of results: The high proportions of brown beans in RCF suggest that this equipment is liable to contribute to the improvement of standard quality of cocoa beans.

ANALISIS SIFAT FISIK BIJI KAKAO PADA BERBAGAI METODE FERMENTASI DAN KONSENTRASI FERMIPAN

This research was done to analyze physical properties of fermented cocoa beans using various fermentation methods and fermipan concentrations. The aim of this study was to identify effective fermentation methods to produce quality cocoa beans. For each trial 10 kg of fresh cocoa beans were treated with fermipan in concentrations 0.5% w/v and 1% w/v. Fermentation trials were conducted in wooden boxes, plastic baskets and in heaps without turning the materials. The trials were conducted aerobically at room temperature for the next 6 days (144 hours). The results showed that the cocoa bean temperature increased significantly the 24th hour of fermentation and then slowly decreased until the end of the fermentation period except for the beans in heaps. The highest temperature of 42oC was found in beans in wooden boxes using the 1% w/v fermipan concentration. During fermentation, the pH of the pulp increased according to the period of fermentation and reached a peak at 144 hour. The highest pH was 7.02 found in beans placed in plastic baskets with 1% w/v of fermipan concentration. Overall the cocoa beans which fermented in wooden boxes with 1% w/v of fermipan gave the best quality with shell content of 15.05 %, brown color intensity of 90%, unfermented beans 2%, insect infestation 1.14% and beans germinated 1.90%.

ANALISIS EKONOMI RUMAH TANGGA PETANI PADA USAHATANI KAKAO RAKYAT DI KABUPATEN BLITAR

As annual crop farmers, the household of cocoa farmers apply diversification livelihood strategy which depends on the season, the labor market, and the free time of each family member to make ends meet. This particular research was conducted in order to analyze the performance and economic behavior of the cocoa farmer households. The research was located in Udanawu and Bakung district with a sample of 45 cocoa farmer households. In order to answer the purpose of this research,a sequences of analysis through tabulation analysis and simultaneous equation models which then estimated using the Two Stage Least Squares (2SLS) method. The results showed that cocoa farming income is significantly influenced by the cocoa prices, production, the number of productive cocoa plants and costs. The expenditure of food on farmer households is significantly affected by the number of the children of the family who are at the time receiving education in school. The most important thing that can be recommended to increase farmer’s income is cocoa bean should be fermented. There needs to be strengthening of institution therefore, the farmers' bargaining position increases and new government policies to push the issuance of mandatory rules for cocoa fermentation.

Effect of aroma potential of Saccharomyces cerevisiae fermentation on the volatile profile of raw cocoa and sensory attributes of chocolate produced thereof

This research describes on-farm-inoculated cocoa bean fermentation with two selected strains of high aroma-producing Saccharomyces cerevisiae isolated from wine fermentation and their effects on volatile profile of resulted raw cocoa material and sensory characteristics of chocolates produced. Both strains produced suitably flavour-active esters, aldehydes, ketones and pyrazines in raw cocoa. Inoculation of S. cerevisiae at 0.5 g kg−1 significantly improved the volatile profile of raw cocoa beans and enabled production of chocolate with enhanced intense fresh fruits and floral notes. Chocolate produced thereof presented better overall quality attributed whatever the fermentation time in comparison to the chocolate issued from spontaneous fermentation. Our results suggested the ways of using selected enhanced yeasts as S. cerevisiae for on-farm implementation of bean fermentation as promising alternative possibilities to improve the cocoa fermentation process, desirable flavourful raw cocoa material and high-aroma-quality chocolates produced.

PTR-QiToF-MS and HSI for the characterization of fermented cocoa beans from different origins

The wide range of geographical cocoa production areas and the increasing consumption trend towards single origin products induced the necessity to verify and certify cocoa beans origin for quality assurance purposes. In this study cocoa beans of various origins were examined by machine olfaction and machine vision techniques. Fifty-nine fermented and dried Forastero cocoa beans from 23 different geographical origins (Africa, Americas, Southeast Asia) were investigated using Proton Transfer Reaction-Quadrupole interface-Time of Flight-Mass Spectrometry and Hyperspectral Imaging to elucidate the geographical information in the beans. The volatile and spectral fingerprints showed the same tendency in clustering samples from Africa separate from those from the Americas. High variability was observed for the Southeast Asian samples, which is most likely related to differences in fermentation. Machine olfaction and machine vision characterization provided a similar degree of separation but are complementary rapid techniques, which may be further developed for use in practical settings.

Development and Characterization of a Pilot-Scale Model Cocoa Fermentation System Suitable for Studying the Impact of Fermentation on Putative Bioactive Compounds and Bioactivity of Cocoa.

Cocoa is a concentrated source of dietary flavanols—putative bioactive compounds associated with health benefits. It is known that fermentation and roasting reduce levels of native flavonoids in cocoa, and it is generally thought that this loss translates to reduced bioactivity. However, the mechanisms of these losses are poorly understood, and little data exist to support this paradigm that flavonoid loss results in reduced health benefits. To further facilitate large-scale studies of the impact of fermentation on cocoa flavanols, a controlled laboratory fermentation model system was increased in scale to a large (pilot) scale system. Raw cocoa beans (15 kg) were fermented in 16 L of a simulated pulp media in duplicate for 168 h. The temperature of the fermentation was increased from 25–55 °C at a rate of 5 °C/24 h. As expected, total polyphenols and flavanol levels decreased as fermentation progressed (a loss of 18.3% total polyphenols and 14.4% loss of total flavanols during fermentation) but some increases were observed in the final timepoints (120–168 h). Fermentation substrates, metabolites and putative cocoa bioactive compounds were monitored and found to follow typical trends for on-farm cocoa heap fermentations. For example, sucrose levels in pulp declined from >40 mg/mL to undetectable at 96 h. This model system provides a controlled environment for further investigation into the potential for optimizing fermentation parameters to enhance the flavanol composition and the potential health benefits of the resultant cocoa beans.

Diversity of yeasts during fermentation of cocoa from two sites in the Brazilian Amazon

ABSTRACT The purpose of this study was to identify the yeasts involved in spontaneous fermentation of cocoa from the Brazilian Amazon region. The fermentation process was carried out experimentally with cocoa seeds from two sites (Medicilândia and Tucumã), State of Pará, northern Brazil, during a six-day period. Totals of 44 yeasts were isolated from Medicilândia and 29 from Tucumã. Molecular identification was carried out by sequencing the D1/D2 region fragment of the rRNA 26S gene, expanded with universal primers for the NL1GC and LS2 eukaryotes. Pichia manshurica and Saccharomyces cerevisiae were identified in Medicilândia and five yeast species (Pichia fermentans, P. kudriavzevii, P. manshurica, S. cerevisiae and Zygosaccharomyces bailii) were identified in Tucumã. The results showed that P. manshurica and S. cerevisiae may have potential for use as starter cultures in future studies to improve the quality of cocoa seeds fermented in the Brazilian Amazon region.

Quality of Chocolate Bar from Fermented Cocoa Beans from Lombok, West Nusa Tenggara

About 500 kgs cocoa beans have been fermented using box fermentation. The fermentation experiments were conducted at the Gangga district North Lombok, West Nusa Tenggara - Indonesia. The starter culture, which contains yeast, lactic acid bacteria, and acetic acid bacteria (107/mL) was added to cocoa beans mass in box fermentation. The fermentation was conducted for 5 days and every 24 hours were mixed. After fermentation, the cocoa beans were dried for 5 days to reduce the moisture content using a sun drying. Dried fermented cocoa beans were processed to be a chocolate bar. The quality of cocoa liquor was determined using organoleptic score. Chocolate aroma, chocolate flavour, and caramelly score were 8.25. The texture and colour score were 8.00 and off flavours score was 0.00. Overall quality of cocoa liquor score was excellent. Analysis of proximate and microbiological properties was carried out to obtain the chocolate bar qualities. The result showed that quality of chocolate bar from fermented cocoa beans appropriate with SNI 7934:2014 about chocolate and chocolate products.

Characteristic of Chocolate Candy Produced from Fermented Cocoa Bean with Lactobacillus plantarum HL-15 Starter Culture for Inhibition Growth of Mycotoxin -Producing Fungi

Characteristic of Chocolate Candy Produced from Fermented Cocoa Bean with Lactobacillus plantarum HL-15 Starter Culture for Inhibition Growth of Mycotoxin -Producing Fungi

Determination of the microbial community in Amazonian cocoa bean fermentation by Illumina-based metagenomic sequencing

High-throughput DNA sequencing (HTS) was used to investigate the microbial community of cocoa bean spontaneous fermentation in the Amazon biome. The characterization was described in three localities from Para state (Amazon biome) and one locality from Bahia state, the top Brazilian state for cocoa production, for comparison of the microbial community structure. The microbial community profile was determined through a metagenomics approach using the Illumina HiSeq platform. Our data showed that the microbial community is composed of a consortium consisting of yeast (i.e., Pichia and Candida), filamentous fungi (i.e., Aspergillus, Penicillium and Fusarium), lactic acid bacteria (i.e., Leuconostoc, Lactococcus, and Lactobacillus), acetic acid bacteria (i.e., Acetobacter and Gluconobacter), enterobacteria and bacilli. Interestingly, Acetobacter pasteurianus was found in higher relative abundance than expected, and three acetic acid bacteria species were identified for the first time in cocoa fermentation (i.e., Komagataeibacter hansenii, Roseomonas sp. and Acidisphaera sp.). Some microorganisms were only detected in Amazonian cocoa and not detected in Bahia cocoa, including six yeast species (i.e., Starmerella bacillaris, Candida temnochilae, Zygosaccharomyces bailii, Candida glabrata, Saccharomycopsis crataegensis and Saccharomycopsis amapae) and seven lactic acid bacteria (i.e., Lactococcus lactis, Fructobacillus pseudoficulneus, Weissella confusa, Pediococcus pentosaceus, Lactobacillus reuteri, Lactobacillus murinus and Lactobacillus pontis). This study provides the first description of the microbiome in cocoa fermentation from the Amazon region, and these data open new perspectives for exploring these microorganisms in biotechnology applications, as well as their use as a starter culture to obtain high-quality cocoa beans.

Cocoa's Residual Honey: Physicochemical Characterization and Potential as a Fermentative Substrate by Saccharomyces cerevisiae AWRI726.

This study aims to characterize the physicochemical properties of cocoa's residual honey and evaluate its fermentative capacity as a substrate, using Saccharomyces cerevisiae AWRI726 as the starter culture for alcoholic fermentation. The research hypothesis was that cocoa's residual honey can be used for the production of fermented beverages. Cocoa's honey has 14.14 g.100 mL−1 of dry material, containing 11.80 g.100 mL−1 of carbohydrates and 1.20% crude protein, in addition to other minor components, such as pectin, lipids, and Fe, Mn, Na, and Zn, with a carbon-to-nitrogen (C/N) ratio (9.8) most suitable for fermentation. Fermentation at 20°C for 240 hours produced a liquid with 16% v/v ethanol (14 g.L−1 in 144 h). However, 24 hours of fermentation produced the maximum ethanol yield (0.373 g.g−1) and volumetric productivity (0.168 g.L−1.h−1), which were associated with a significant increase in the specific cell growth rate. Saccharomyces cerevisiae AWR1726 performed satisfactorily in the alcoholic fermentation of cocoa's residual honey, similar to that observed in other fruit beverages, thus suggesting the suitability of cocoa's residual honey for future commercial applications.

Probiotic Potential, Antioxidant Activity, and Phytase Production of Indigenous Yeasts Isolated from Indigenous Fermented Foods.

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 pH2.0, 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.