Fermented Cocoa Beans Research Articles

Quality Assessment During the Fermentation of Cocoa Beans: Effects of Partial Mucilage Removal

Fermentation of cocoa beans is the most important process contributing to the flavor in chocolate and other related products. The present study aimed to investigate the fermentation at a laboratory scale of cocoa beans with and without 10% w/w mucilage removal (whole beans). The physicochemical properties and microorganism development were monitored for six days of continuous fermentation (sampling was conducted every 24 hours). The results indicated the effects of partial mucilage removal of cocoa beans before the fermentation, in which the temperature, pH, and mucilage content (with/without mucilage removal) were recorded as 36.5 oC/38.6 oC, 3.44/3.31, and 18.41%/21.84%, respectively at the final day. Besides, the density of microorganisms (yeast-mold, lactic acid bacteria, and acetic acid bacteria) of cocoa beans with partial mucilage removal was higher than whole cocoa beans due to the increased aeration of the beans with mucilage removal, creating favorable conditions for the growth of microorganisms. After the fermentation, several physicochemical properties of the two cocoa bean types were compared, which demonstrated the more favorable quality of the cocoa beans with partial mucilage removal compared to the whole cocoa beans for the fermentation, e.g., lower seed shell content (14.1% vs. 17.8%), lower total acid (1.67% vs. 2.77%), and pH of around 5.0.

Hedonic Test of Pegagan Chocolate Products at PT XYZ

Handling of raw materials for cocoa beans is an important process to maintain the quality of the processed chocolate. There are several stages carried out in the post-harvest cocoa process including fruit picking, fruit breaking, cocoa bean fermentation, drying cocoa beans and storage. The purpose of this study was to determine whether or not a sensory assessment of the preference for pegagan chocolate was made, and to analyze the panellists’ level of acceptance of pegagan chocolate. Pegagan leaf chocolate is produced with the aim of providing good nutrition, brain multivitamins and memory enhancers. Sensory assessment of the hedonic test using 20 untrained panellists from the parameters measured were color, taste, texture, aroma and shape. The panellists’ acceptance of chocolate with the addition of pegagan leaves was that the majority liked the chocolate on all parameters.

Saccharomyces cerevisiae and Pichia manshurica from Amazonian biome affect the parameters of quality and aromatic profile of fermented and dried cocoa beans.

The use of yeasts as starter cultures is a promising alternative to produce fermented cacao with particular characteristics regarding the quality of aromas and physical and chemical characteristics that are accepted by the chocolate market. This study aimed to evaluate the physical and chemical transformations of cocoa beans during fermentation after inoculation with starter cultures of yeast species Pichia manshurica (PF) and Saccharomyces cerevisiae (SF), both previously isolated in cocoa bean fermentations in the Brazilian Amazon, in comparison with a fermentation without the inoculum addition (CF). During the fermentation time, which was carried out on a cocoa farm in Igarapé-Miri (Amazon biome, Pará, Brazil), the contents of phenolic compounds (catechin and epicatechin), sugars (glucose, fructose, and sucrose), acetic acid, and ethanol were monitored by HPLC, and the volatile compounds profiles were assessed by GC-MS. The starter culture of P. manshurica was able to produce fermented cocoa beans with highly desirable characteristics for the production of good quality chocolate: low acidity, a broad variety of aromatic compounds with floral, fruity, and sweet characteristics, in addition to showing high contents of catechin and epicatechin, which are known by their antioxidant properties. Therefore, the use of starter cultures with species of yeasts isolated in the Amazon region, during cocoa fermentation, is an alternative to obtain fermented seeds with high quality favoring the commercial agreements in the chocolate market by cocoa producers. PRACTICAL APPLICATION: The addition of starter cultures was able to produce cocoa beans with good quality. Yeasts species isolated and identified in Amazonian cocoa fermentation can improve the profiles of aromatic compounds. Catechin and epicatechin contents are higher in inoculated cocoa beans fermentations.

Perbandingan Kualitas Fermentasi Biji Kakao dengan Menggunakan Kultur Campur dan Kultur Tunggal Saccharomyces cerevisiae

Cocoa fermented products in Indonesia generally produce poor quality fermentation. This study was conducted to compare the quality of fermented cocoa beans with the addition of mixed and single cultures of Saccharomyces cerevisiae. This study used a randomized block design (RAK) method with 7 treatments, namely control, single culture (KT5%), (KT10%), (KT15%) and mixed culture (KC5%), (KC10%), (KC15%). Observation parameters include pH, temperature, water content, reducing sugar content, lactic acid content, acetic acid content, total fat content, cut test and organoleptic. The analytical data obtained were then analyzed with the SAS device with a further test of 0.05% BNJ. The results showed that the addition of mixed culture (KC15%) had an effect on the quality of fermented cocoa beans. The quality of fermented cocoa beans in KC15% treatment showed reducing sugar content of 6.51%, lactic acid content of 0.05%, acetic acid content of 0.79%, total fat content of 50.10%, fermented beans 78% and organoleptic results on texture. , color, aroma and distinctive taste of cocoa, resulted in a good assessment of the KC15% treatment.

Yeasts as Producers of Flavor Precursors during Cocoa Bean Fermentation and Their Relevance as Starter Cultures: A Review

During the fermentation of cocoa beans, the yeasts produce volatile organic compounds (VOCs). Through reactions associated with amino acid metabolism, yeasts generate important aroma precursors as acetate esters and fatty acid ethyl esters are essential in developing fruity flavors and aromas in the final product (usually chocolate). In addition, some yeasts may have pectinolytic and antifungal activity, which is desirable in the post-harvest process of cocoa. The main yeast species in cocoa fermentation are Saccharomyces cerevisiae, Pichia kudriavzevii, and Hanseniaspora opuntiae. These produce higher alcohols and acetyl-CoA to make acetate–esters, compounds that produce floral and fruity notes. However, there are still controversies in scientific reports because some mention that there are no significant differences in the sensory characteristics of the final product. Others mention that the fermentation of cocoa by yeast has a significant influence on improving the sensory attributes of the final product. However, using yeasts as starter cultures for cocoa bean fermentation is recommended to homogenize sensory attributes such as notes and flavors in chocolate.

Effect of spontaneous fermentation location on the fingerprint of volatile compound precursors of cocoa and the sensory perceptions of the end-chocolate.

Cocoa pod-opening delay and bean fermentation promote the organoleptic quality of chocolate. The present research investigated the changes in the volatile fingerprint of cocoa harvested at a traditional plantation. Cocoa beans extracted from 2-days pod-opening delay were simultaneously fermented for 5days using container and then sun-dried to 7-8% moisture content at five different locations: Akoupé, San Pedro, Soubré, Djekanou and Daloa. The aromatic analysis were done on cocoa using the HS-SPME-GC/MS technique. Professional panelists evaluated the sensory perceptions of the chocolate. The results shows that cocoa fermented in both Daloa and Soubré regions were differentiated by 2,3-butanediol while those processed in other regions presented highest acetoin content. However, fermented cocoa from Soubré region exhibited most amount of 2,3-butanediol, diacetate A whereas 2,3,5,6-tetramethylpyrazine differentiated those from Daloa region. Sensory properties of chocolate were not linked to the aromatic compound precursors profile of beans. The fermentation performed in San Pédro region promote both the generation of more desirable aromatic compounds of cocoa and sensory attributes of the finished chocolate. The fermentation location generates a greater differentiation of the volatile fingerprint of cocoa and the sensory perceptions of the finished chocolate.

Unique metabolite profiles of Indonesian cocoa beans from different origins and their correlation with temperature

Chocolate flavors vary depending on the origin of the cocoa beans used. Differences in soil, microorganisms, and environmental factors contribute to the formation of flavor precursors in cocoa beans. During cocoa bean fermentation, environmental temperature has been shown to alter metabolite concentrations. However, the correlation between the metabolite profile of cocoa beans and the temperature of their region of origin has not been fully defined. In this study, the metabolite profiles of Indonesian cocoa beans of various origins were evaluated using gas chromatography/mass spectrometry-based analysis, and were found to differ depending on the origin of the bean. Subsequently, the correlation between metabolite profile and environmental temperature of the origin was assessed using orthogonal projection to latent structure regression (OPLS-R) analysis. The analysis revealed that seven metabolites were associated with temperature: γ-aminobutyric acid (GABA), ethanolamine, glycerol, isocitric acid+ citric acid, succinic acid, malic acid, and saccharic acid. The findings of this study will be valuable to chocolate industries for the production of single-origin chocolates.

Identification of Lactic Acid Bacteria on Raw Material for Cocoa Bean Fermentation in the Brazilian Amazon

The prevalent microbiota during cocoa fermentation is one of the main factors responsible for the chemical and biochemical changes that lead to desirable chocolate characteristics. However, the microbiota can be affected by several factors, including the edaphoclimatic conditions, which are typical of the production region. The objective of this study was to identify LAB in Amazonian cocoa prior to fermentation. LAB were isolated using MRS agar. By sequencing the 16S region of isolated LAB in MRS agar, it was possible to identify five LAB species that were registered in the GenBank database (accessions from MT117900 to MT117915). They included Pediococcus acidilactici, which was the most prevalent, followed by Lactobacillus farraginis, L. parafarraginis, L. zeae, and L. casei. Studies are needed to ascertain their specific roles and impact on cocoa quality. It is likely that they can be available as starter cultures to enhance the quality characteristics of chocolate.

Non‐conventional fermentation at laboratory scale of cocoa beans: Using probiotic microorganisms and substitution of mucilage by fruit pulps

SummaryIn a non‐conventional lab‐scale fermentation of cocoa beans using probiotic microorganisms and substituting the cocoa pulp for fruit pulp, physicochemical, microbiological and quality parameters were investigated. Two hundred grams of beans were fermented in a controlled environmental chamber (temperature ramp rate of 25°C for 48 h; 35°C for 48 h and 45°C for 48 h; and 65% HR). pH, titratable acid, citric, lactic and acetic acids, as well as sugars and ethanol were measured. A cut test was also performed on the cocoa beans fermented 5 and 6 days. As the fermentation time progressed, citric acid concentration decreased until 0.53 g kg−1, whereas both lactic and acetic acids increased until 0.44 and 16.58 g kg−1, respectively. Sucrose content decreased from 12.26 g kg−1 (in fresh) to 6.54 g kg−1 on the 6th day. Fructose and glucose contents increased in the cotyledons from day five, reaching a maximum concentration of 1.14 and 1.01 g kg−1, respectively, on day six. Yeasts were the main microorganisms during the first 24–48 h (8.4 log CFU g−1), while bacterial counts reached its highest number (7.8 log CFU g−1) on day four. Beans fermented 5 and 6 days resulted in more fermented beans (>81%) and less violet ones (<18.4%) than the control.

Fermentation Practices and Microbiological Profile of Fermented Cacao Beans (Theobroma cacao) from Mvila Division, South Region, Cameroon

Fermentation is an important process in the production of commercial cocoa beans. Conducted in a natural way, it takes place through the action of microorganisms such as lactic acid bacteria (LAB), acetic bacteria and fungi. This generally contributes to obtaining cocoa of different grades. In order to improve the quality of commercial cocoa in the division of Mvila, the aim of this study was to assess the fermentation practices and the microbiological profile of fresh fermented cocoa beans. For this purpose, a survey on the fermentation techniques of the cocoa sector was firstly done with 75 producers. Thereafter, samples of fermented and dried cocoa beans from different producers were collected for commercial quality assessment (grades, impurities rate and water content). Producers with cocoa beans of good quality were retained and the microbial (LAB, yeast enumeration and characterization) quality of their fresh fermented cocoa were monitored after fermentation. The results obtained on the post-harvest treatment techniques of cocoa beans revealed that the technical itinerary is respected with 100% of producers who put the pots in incubation before fermentation. In addition, fermentation techniques are practiced by 87,5% of producers and take place for 6 days long (in 95 %), notably fermentation in wooden box (47.5%), fermentation in heaps (32.50%). According to the commercial quality, the localities of Melane and Nkoemvone have good quality (Grade=1) cocoa with a rate of brown beans of 95.4% and 94% respectively. Lactic acid bacteria, acetic bacteria and yeast were presents in all fresh cocoa beans with the final load of 1.17 × 107 CFU/g, 1.87 × 105 CFU/g and 5.60 × 106 CFU/g respectively. The fermentative capacity revealed that among the 13 isolates of yeasts, 11 were alcohol producers. This study shows that these microorganisms are responsible for the good fermentation of cocoa beans. It would be judicious to make molecular identification of these strains and to use them as a starter for the controlled fermentation of cocoa.

Temperature and pH Monitoring System Design in the Fermentation of Cocoa Beans Based on Android

The fermentation of cocoa beans is a key step in increasing their quality. Temperature and pH have an impact on the fermentation process' performance. Until, temperature and pH measurements in the cocoa bean fermentation process have been done with a thermometer and a pH meter, and have been heavily impacted by experience and knowledge. The goal of this research is to create a temperature and pH monitoring system for the fermentation of cocoa beans using an android platform. The sensor must be able to measure temperature and pH during the fermentation process, and the sensor reading data must be displayed on the android display, according to the system design criteria. A temperature sensor, pH sensor, microcontroller, TP-Link or WiFi modem, ThingSpeak (data storage), and Smartphone make up the system's hardware. Arduino CC software version 1.8.14 was used to create the software. The system hardware was made up of numerous components, including the NodeMCU ESP32 microcontroller, SKU SEN0161 pH sensor, DS18B20 temperature sensor, 4.2k resistor, jumper cable, and 5 volt charger adaptor, according to the results. A plastic container houses the electronic system circuit. The monitoring system can read the temperature and pH during the fermentation process, which lasts 112 hours or 6 days, according to the performance test findings. During fermentation, the temperature of cocoa beans ranged from 27.19 °C to 41.75 °C, with the pH fluctuating between 2.50 and 7.10.

Improving Data Security with Blockchain and Internet of Things in the Gourmet Cocoa Bean Fermentation Process.

Brazil was one of the largest cocoa producers in the world, mainly supported by the South of Bahia region. After the 1980s, the witch-broom disease demolished plantations, and farmers were forced into bankruptcy. The worldwide search for gourmet cocoa has rekindled interest in this production, whose fermentation process is a key step in obtaining fine cocoa, thanks to the fact that many processing properties and sensory attributes are developed in this phase. This article presents a blockchain-IoT-based system for the control and monitoring of these events, aiming to catalog in smart contracts valuable information for improvement of the cocoa fermentation process, and future research. Blockchain is used as a distributed database that implements an application-level security layer. A proof of concept was modeled and the performance of the emulated system was evaluated in the OMNet simulator, where a technique based on the SNMP protocol was applied to reduce the amount of data exchanged and resources served/consumed in this representation. Then, a physical platform was developed and preliminary experiments were performed on a real farm, as a way to verify the improvement of the cocoa fermentation process when using a technological approach.

Comparison of the Aroma Composition and Sensory Properties of Dark Chocolates Made with Moist Incubated and Fermented Cocoa Beans.

In a previous investigation, "moist incubation" was described as a novel postharvest treatment for cocoa and the aroma composition of the resulting cocoa nibs was compared to unfermented and fermented cocoa nibs. For this treatment, unfermented and dried nibs are rehydrated with an aqueous solution containing lactic acid and ethanol to adjust the pH value and are subsequently incubated at 45 °C under aerobic conditions for 72 h before drying. The aim of the present study was to investigate the sensory properties and aroma composition of dark chocolates made of these materials after roasting. Therefore, gas chromatography-olfactometry (GC-O) in combination with aroma extract dilution analysis (AEDA), quantitation with isotopically labeled standards, odor activity value (OAV) determination, and sensory analysis were performed. The three different chocolates had distinct sensory and OAV profiles. The sensory profiles showed a higher intensity of fruity aroma notes and a lower intensity of bitterness and astringency in the chocolate made with the moist incubated cocoa, while the chocolate made of fermented cocoa reached higher scores in the roasty aroma notes. Furthermore, higher OAVs were determined for the Strecker aldehydes in the chocolate made of the moist incubated cocoa, whereas higher OAVs for the pyrazines and the acids were detected in the chocolate made of fermented cocoa. In contrast, the chocolate produced with the unfermented cocoa showed low cocoa specific aroma notes and high levels of astringency and bitterness. The detected differences reveal interesting insights into the influence of different postharvest treatments on the resulting aroma composition in the final chocolate. Furthermore, the alternative postharvest treatment was demonstrated to result in chocolates with a pleasant sensory profile.

Acetobacter senegalensis isolated from mango fruits: Its polyphasic characterization and adaptation to protect against stressors in the industrial production of vinegar: A review.

It has been more than a decade since Acetobacter senegalensis was isolated, identified and described as a thermotolerant strain of acetic acid bacteria. It was isolated from mango fruits in Senegal and used for industrial vinegar production in developing countries, mainly in sub-Saharan Africa. The strain was tested during several spirit vinegar fermentation processes at relatively high temperatures in accordance with African acclimation. The upstream fermentation process had significant stress factors, which are highlighted in this review so that the fermentation process can be better controlled. Due to its high industrial potential, this strain was extensively investigated by diverse industrial microbiologists worldwide; they concentrated on its microbiological, physiological and genomic features. A research group based in Belgium proposed an important project for the investigation of the whole-genome sequence of A. senegalensis. It would use a 454-pyrosequencing technique to determine and corroborate features that could give this strain significant diverse bio-industrial applications. For instance, its application in cocoa bean fermentation has made it a more suitable acetic acid bacterium for the making of chocolate than Acetobacter pasteurianus. Therefore, in this paper, we present a review that summarizes the current research on A. senegalensis at its microbial and genomic levels and also its specific bio-industrial applications, which can provide economic opportunities for African agribusiness. This review summarizes the physiological and genomic characteristics of Acetobacter senegalensis, a thermotolerant strain isolated from mango fruits and intended to be used in industrial vinegar fermentation processes. It also explores other bio-industrial applications such as cocoa fermentation. Vinegar fermentation is usually performed with mesophilic strains in temperate regions of the world. Developing countries, such as Senegal, import vinegar or make 'fake' vinegar by diluting acetic acid obtained from petrochemicals. The use of a thermotolerant Acetobacter senegalensis strain as a solid functional starter culture, as well as the design of a new adapted bioreactor, has significantly contributed to food security and the creation of small- to medium-sized enterprises that produce mango vinegar in West Africa.

PENINGKATAN MUTU KAKAO MELALUI FERMENTASI MENGGUNAKAN STARTER KERING BAKTERI ASAM LAKTAT DAN BAKTERI ASAM ASETAT INDIGENUS KAKAO ACEH

Indonesia is one of the cocoa producing countries in the world. Although the amount of cocoa production and planted area has increased, the quality of cocoa beans in Indonesia including Aceh is still low. This happened because the farmers did not carry out the fermentation process. Fermentation is one of the most important steps that must be taken to improve cocoa quality. However, it is rarely done because it takes a long time. The purpose of this study was to analyze the use of starter bacteria lactic acid (LAB) and acetic acid (BAA) indigenous Aceh cocoa in fermenting Aceh cocoa. The research was conducted by fermenting cocoa with two different factors, namely the type of starter (control, LAB, BAA, LAB-BAA mixture) and fermentation time (3,4 and 5 days). The results showed that the use of Acehnese cocoa indigenus bacteria starter and different fermentation time had an effect on fermentation conditions and the quality of fermented cocoa beans. Fermentation of cocoa beans with the addition of a dry starter affects the pH value and the percentage of fermented cocoa beans. The best treatment was obtained when using LAB starter with fermentation time of 4 days, with the same fermented quality of cocoa beans with fermentation using LAB and BAA for 5 days of fermentation. Further research is needed to confirm the quality of fermented cocoa beans produced by analyzing volatile chemical components and flavors.

Exploring cocoa bean fermentation mechanisms by kinetic modelling.

Compared with other fermentation processes in food industry, cocoa bean fermentation is uncontrolled and not standardized. A detailed mechanistic understanding can therefore be relevant for cocoa bean quality control. Starting from an existing mathematical model of cocoa bean fermentation we analyse five additional biochemical mechanisms derived from the literature. These mechanisms, when added to the baseline model either in isolation or in combination, were evaluated in terms of their capacity to describe experimental data. In total, we evaluated 32 model variants on 23 fermentation datasets. We interpret the results from two perspectives: (1) success of the potential mechanism, (2) discrimination of fermentation protocols based on estimated parameters. The former provides insight in the fermentation process itself. The latter opens an avenue towards reverse-engineering empirical conditions from model parameters. We find support for two mechanisms debated in the literature: consumption of fructose by lactic acid bacteria and production of acetic acid by yeast. Furthermore, we provide evidence that model parameters are sensitive to differences in the cultivar, temperature control and usage of steel tanks compared with wooden boxes. Our results show that mathematical modelling can provide an alternative to standard chemical fingerprinting in the interpretation of fermentation data.

Screening Wild Yeast Isolated from Cocoa Bean Fermentation Using Volatile Compounds Profile.

Yeasts are one of the main ingredients responsible for flavor precursors production associated with sensorial characteristics in chocolate. Using wild yeast isolated from cocoa beans fermentation is emerging as a strategy for developing starter cultures. However, the volatile compounds (VCs) produced by yeasts are not yet known. This study aimed to select wild yeasts with the potential to produce volatile compounds associated with desirable flavor attributes. A total of 150 wild yeasts strains were isolated from the spontaneous cocoa beans fermentation, of which 40 were identified by morphology and physiological features. VCs produced were identified and quantified using SPME-GC-MS and GC-FID and profiles were evaluated statistically by PCA and cluster analysis for the compounds that had a high odor threshold value. Thirty-six VCs produced by these yeasts were identified into six main families, namely esters, alcohols, acids, aldehydes, ketones, and pyrazines. PCA showed the separation of the yeasts into two main clusters. Strains, Y195 and Y246, belong to the first cluster and are the highest producers of alcohols related to floral perceptions. In the second cluster, thirty-three yeasts were grouped by their ability to produce esters. Of all of them, Y110MRS stood out for producing 2-phenyl ethyl acetate and isoamyl acetate associated with fruity perceptions. This screening allowed us to identify yeasts that produced VCs of technological interest and which could be used to develop a starter culture.

Cocoa bean fingerprinting via correlation networks

Cocoa products have a remarkable chemical and sensory complexity. However, in contrast to other fermentation processes in the food industry, cocoa bean fermentation is left essentially uncontrolled and is devoid of standardization. Questions of food authenticity and food quality are hence particularly challenging for cocoa. Here we provide an illustration how network science can support food fingerprinting and food authenticity research. Using a large dataset of 140 cocoa samples comprising three cocoa fermentation/processing stages and eight countries, we obtain correlation networks between the cocoa samples by computing measures of pairwise correlation from their liquid chromatography-mass spectrometry (LC-MS) profiles. We find that the topology of correlation networks derived from untargeted LC-MS profiles is indicative of the fermentation and processing stage as well as the origin country of cocoa samples. Progressively increasing the correlation threshold firstly reveals network clusters based on processing stage and later country-based clusters. We present both, qualitative and quantitative evidence through network visualization, network statistics and concepts from machine learning. In our view, this network-based approach for classifying mass spectrometry data has broad applicability beyond cocoa.

A method for non-destructive determination of cocoa bean fermentation levels based on terahertz hyperspectral imaging

Fermentation of cocoa is a key process to obtain aromatic chocolate products from raw cocoa beans. Hitherto, the levels of fermentation in cocoa are determined using destructive techniques, for example by a cut-test to manually observe the colour inside the beans, or by quantifying ammonia nitrogen (NH3) in the cocoa powder. In this paper, we present the use of Terahertz hyperspectral imaging as a new way to non-destructively analyse and detect fermented cocoa beans. The study analysed two sets of twenty-two cocoa bean samples with different levels of fermentation from two producers in Brazil. A correlation between fermentation conditions and the outcome results of their THz measurements was observed.

Ecology and population dynamics of yeast starter cultures in cocoa beans fermentation.

This study aimed to investigate controlled fermentation of cocoa beans with selected yeasts as starter cultures via integrating microbiological, biochemical, and chromatographic analyses. The steps involved in the yeast starter culture test were of the following order: 1) counting, isolation, purification, and biochemical identification of yeasts, 2) selection of ethanol-producing yeasts, 3) selection of thermotolerant yeasts, and 4) evaluation of physicochemical parameters of the selected yeasts in controlled fermentation of cocoa (F1 - Saccharomyces ssp. and Hanseniaspora ssp. and F2 - spontaneous fermentation - control). A total of 32 yeasts were isolated from three sampling points (M1, M2, and M3), which comprised 50% Candida ssp., 9.4% Rhodotorula ssp., 18.8% Saccharomyces ssp., and 18.8% Hanseniaspora ssp. The yeasts identified as Saccharomyces ssp. (n = 6) were subjected to the ethanol production test. Saccharomyces spp. CLV09 showed the highest concentration of ethanol in the simulated cocoa medium (3.5% v/v). Hanseniaspora spp. CVL20 and CVL19 strains showed the highest thermotolerance at 42°C after 72 h of growth. The starter cultures with Saccharomyces ssp. and Hanseniaspora ssp. showed a similar growth rate of the mesophilic aerobic population in both F1 and F2. Fermentation of the starter culture showed a higher production of organic acids than spontaneous fermentation (F2). Thus, Saccharomyces ssp. and Hanseniaspora ssp. can be used as a starter culture in cocoa fermentation.