Fermented Cocoa Research Articles

Variations in Ecuadorian Cocoa Fermentation and Drying at Two Locations: Implications for Quality and Sensory.

In Ecuador, various processes are applied during cocoa post-harvesting. This study, therefore, explored fermentation parameters across two locations with 2-7 independent runs, focusing on temperature, microbial counts, pH during fermentation and drying, and their impact on cocoa bean quality. Factors including fermentation devices (jute bags, plastic bags, and wooden boxes), pre-drying, turning during fermentation, fermentation duration, and drying temperature were investigated. Fermenting in plastic bags without pre-drying or turning and fermenting in jute bags for only 40 ± 2.0 h yielded low maximal fermentation temperatures Tmax (31.1 ± 0.4 °C and 37.6 ± 1.8 °C), leading to bitter, astringent, woody, and earthy cocoa liquor. Longer fermentation (63 ± 6 h) in wooden boxes with turning (Wt) and in jute bags with pre-drying and turning (Jpt) achieved the highest Tmax of 46.5 ± 2.0 °C, and a more acidic cocoa liquor, particularly in Wt (both locations) and Jpt (location E). Therefore, it is recommended to ferment for a minimum duration from day 1 to 4 (63 ± 6 h), whether using plastic bags (with mandatory pre-drying) or jute bags (with or without pre-drying or turning). Furthermore, this study underscores the risks associated with excessively high drying temperatures (up to 95.2 ± 13.7 °C) and specific dryer types, which can falsify cut-tests and introduce unwanted burnt-roasted off-flavors in the cocoa liquor.

PHYSICAL AND CHEMICAL CHARACTERICSTICS OF SULAWESI COCOA BEANS 2 (Theobroma cacao L) FERMENTED AND UNFERMENTED RESULT

This study aims to determine the physical and chemical characteristics of fermented and unfermented BR 25 clone dry cocoa beans. The treatment in this study was the fermented treatment (Treatment A) and the unfermented treatment (Treatment B). The fermented and unfermented treatments were processed from wet cocoa beans with 10 kilograms of beans for each treatment. The fermentation treatment uses a box of Styrofoam, while the unfermented treatment is directly dried in the sun after removing the mucus (pulp). The parameters for fermented and unfermented dry cocoa beans were water, pH, fat content, protein, polyphenols, antioxidant activity (IC50), and ALT. The results showed that fermented cocoa beans had a moisture content of 5.77%, pH of 6.41; fat content of 55.63%; protein of 14.36%; polyphenol content of 4.62%; antioxidant activity (IC50) of 21.96%; and ALT value of 1.0 x 102 colonies/gram. Unfermented cocoa beans have a moisture content of 6.52%, pH of 5.20; fat content of 59.63%; protein of 17.83%; polyphenol content of 2.06%; antioxidant activity (IC50) of 62.23%; and ALT value of 1.1 x 102 colonies/gram.Keywords: cacao beans; fermented; unfermented; characteristics

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

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

Evaluation of a cocoa dryer prototype using LabVIEW software and Arduino hardware

In a country like Colombia, food drying plays an important role in the extension of technology so that farmers improve the quality of their productivity. The objective of this work is to develop a system for acquiring thermodynamic variables, to establish the energy efficiency of a product drying oven, specifically for cocoa beans using Arduino hardware and LabView software. This research begins by establishing the variables to be measured and recorded by the DHT11 sensors, such as temperature and humidity. These sensors are connected to an Arduino Uno board that has an Atmel microcontroller, which captures the information from each of these. An algorithm was also developed that captures the temperature and humidity data and sends it through the serial port to the LabView software, in which the visualization on the front panel and programming in the block diagram have been developed, being viewed from the HMI (Human Machine Interface). For the experimental tests, 3,309 grams of fermented cocoa beans were taken, and dried for 36 hours, removing a total mass of water of 1,650 grams. The results show an energy efficiency of 10.62%, concluding that the drying oven that takes advantage of the residual heat of an 18000 BTU condenser integrated with the proposed variable system is suitable for drying cocoa beans. Despite this efficiency being low, it meets the objective of removing moisture from them.

Palatability Test of Urea Molasses Multinutrient Block (UMMB) Based on Cocoa Pods Fermented with LMO (Local Microorganisms) in Sheep

This study aims to determine the benefits of giving UMMB (Urea molasses Multinutrient Block) based on cocoa fruit peels fermented by local microorganisms (LMO). The research was conducted at Dompet Dhuafa farm, Bulu Cina village, Hamparan Perak district, Deli Serdang. The research was conducted using a completely randomized design (CRD) with 3 treatments and 3 replications. The three treatments are K1 = UMMB containing 10% unfermented cocoa pods; K2 = UMMB containing 10% fermented cocoa pods; and K3 = UMMB containing 15% fermented cocoa pods. The parameters observed were palatability, feed consumption, feed conversion, and body weight gain. The results showed that UMMB (Urea molasses Multinutrient Block) based on fermented cocoa fruit peel with LMO did not significantly affect UMMB palatability, feed consumption, body weight gain, and feed conversion of sheep.

Fermentasi Kakao di Kelompok Tani Plea Puli Desa Bloro Kabupaten Sikka

Bloro Village is one of the villages in Nita District, Sikka Regency, East Nusa Tenggara Province with the majority of the population working as farmers. The leading commodity in this village is Cocoa. The Plea Puli Farmers Group is one of the farmer groups involved in cocoa cultivation, including the seeding and processing of the results. The Plea Puli Farmers Group has a chocolate industrial house with production products in the form of cocoa powder and chocolate bars. To get a quality chocolate bar and cocoa powder products, you need to pay attention to post-harvest handling, namely cocoa fermentation. Because it is at this stage that determines the precursor to the distinctive taste of cocoa. However, the fact is that most farmers do not ferment properly. This program aims to provide education to farmers about post-harvest handling, especially cocoa fermentation. Through this activity, it is hoped that farmers can understand that to obtain good quality cocoa beans, the thing that must be considered is post-harvest handling, especially cocoa fermentation. This program begins with a discussion with several members of the Plea Puli Farmers Group and the preparation of an activity plan with the farmer group. Followed by the preparation stage, namely preparing tools and materials. At the implementation stage, activities began with socialisation about cocoa fermentation, then continued with harvesting, fruit sorting, ripening, fruit splitting, wet bean sorting, fermentation, and drying activities with the Plea Puli Farmers Group.

Investigating luxS gene expression in lactobacilli along lab-scale cocoa fermentations

Previous metagenomic analyses have suggested that lactobacilli present potential for Quorum Sensing (QS) in cocoa fermentation, and in the present research, laboratory scale fermentations were carried out to monitor the expression of luxS, a universal marker of QS. For that, 96 h-fermentations were studied, as follows: F0 (non inoculated control), F1 (inoculated with yeasts, lactic acid bacteria, and acetic acid bacteria), F2 (inoculated with yeasts and acetic acid bacteria), F3 (inoculated with yeasts only). The parameters evaluated were: plate counting, quantification of key enzymes and analysis of volatile organic compounds associated with key sensory descriptors, using headspace gas chromatography-mass spectrometry (GC-MS). Furthermore, QS was estimated by the quantification of the expression of luxS genes by Reverse Transcriptase Real-Time PCR. The results demonstrated that microbial succession occurred in pilot scale fermentations, but no statistical differences for microbial enumeration and α-diversity index were observed among experiments and control. Moreover, it was not possible to make conclusive correlations of enzymatic profile and fermenting microbiota, likely due to the intrinsic activity of plant hydrolases. Regarding to the expression of luxS genes, in Lactiplantibacillus plantarum they were active along the fermentation, but for Limosilactobacillus fermentum, luxS was expressed only at early and middle phases. Correlation analysis of luxS expression and production of volatile metabolites evidenced a possible negative association of Lp. Plantarum with fermentation quality. In conclusion, these data corroborate former shotgun metagenomic analysis by demonstrating the expression of luxS by lactobacilli in pilot scale cocoa fermentation and evidence Lp. Plantarum is the main lactic acid bacteria related to its expression.

Effectiveness of public service program related cocoa fermentation sop in increasing the knowledge of cocoa farmer

Cocoa farmers in this area were not familiar with post-harvest processing, usually, the cocoa bean was sold directly without any fermentation. This public service objective provides information on good quality control of cocoa plant beans, such as fermentation pH, sorting, and contained water content. Followed to improve the knowledge about fermented cocoa bean's characteristics, especially about fine flavor cocoa (FFC) which has a flavor profile and also several active compounds for healthy. Data collection was carried out by filling out questionnaires from 25 farmer respondents. The results of the service showed that as many as 52% of the respondents have been cocoa farmers for more than 5 years. Most of the cocoa farmers produced more than 500 kg/harvest. Many farmers do not ferment cocoa beans because the fermentation process takes a long time, while farmers want to sell cocoa beans quickly. This causes the selling price of cocoa beans to become low.

Viabilitas mikroba selulolitik pada media pod dan pulp kakao (Theobroma cacao L.)

Cocoa production in Indonesia is increasing from year to year. Cocoa production increased has an impact on increasing waste cocoa pod and pulp. Cocoa pod contains lignocellulose, while cocoa pulp contains many sugars. The composition of cocoa pod and pulp allows the cellulolytic microbe to grow. This study aims to perform microbial selection from fermented cocoa and sugar cane can grow in cocoa pod and pulp (Theobroma cacao). The selection method using the CMC 1%, pod 1%, pulp 1%, and pod 0,5%+pulp 0,5% media. Then, the analysis of reducing sugar using DNS method, while the total sugar analysis using phenol-H2SO4 method. Sugars of hydrolisis result were analyzed quantitatively using TLC. The results of microbial selection on CMC 1%, pod %, pulp 1%, and pod 0,5%+pulp 0,5% media obtained TBT 3.2 have the largest cellulolytic index on the pod 1% media. Isolates TBT 3.2 grown on pod 1% medium produce reducing sugar is 4,965 mg/mL at 30 hours sampling with highest total sugar at 6 hours sampling is 9,789 mg/mL. The types of sugars identified using the TLC are mannose, galactose, glucose, and selobiose.

Viabilitas mikroba selulolitik pada media pod dan pulp kakao (Theobroma cacao L.)

Cocoa production in Indonesia is increasing from year to year. Cocoa production increased has an impact on increasing waste cocoa pod and pulp. Cocoa pod contains lignocellulose, while cocoa pulp contains many sugars. The composition of cocoa pod and pulp allows the cellulolytic microbe to grow. This study aims to perform microbial selection from fermented cocoa and sugar cane can grow in cocoa pod and pulp (Theobroma cacao). The selection method using the CMC 1%, pod 1%, pulp 1%, and pod 0,5%+pulp 0,5% media. Then, the analysis of reducing sugar using DNS method, while the total sugar analysis using phenol-H2SO4 method. Sugars of hydrolisis result were analyzed quantitatively using TLC. The results of microbial selection on CMC 1%, pod %, pulp 1%, and pod 0,5%+pulp 0,5% media obtained TBT 3.2 have the largest cellulolytic index on the pod 1% media. Isolates TBT 3.2 grown on pod 1% medium produce reducing sugar is 4,965 mg/mL at 30 hours sampling with highest total sugar at 6 hours sampling is 9,789 mg/mL. The types of sugars identified using the TLC are mannose, galactose, glucose, and selobiose.

ANALYSIS OF COCOA FERMENTATION WITH THE ADDITION OF YEAST AND LACTIC ACID BACTERIA

Abstract One of the factors that affect the quality of cocoa beans is post-harvest handling such as the fermentation process. Fermentation plays an important role in determining the final quality of dried cocoa beans. Chocolate flavors can be produced from microbial fermentation from high-quality cocoa bean raw materials by utilizing microbes. This study aims to calculate the number of microbes at the beginning and end of the cocoa bean fermentation process, observing the pH value, temperature, and color of cocoa beans. The treatment carried out is a fermentation variation consisting of: (1) natural fermentation or without the addition of microorganisms; (2) controlled fermentation with the addition of microorganisms consisting of Saccharomyces cerevisiae, Lactic Acid Bacteria simultaneously added at the beginning of fermentation. The addition of microorganisms is 106 CFU / 1kg of cocoa beans each. Fermentation is carried out in a fermentation box with a capacity of 1 kg of fresh cocoa, at room temperature (33-35oC), for 4 days. The analysis carried out includes calculating the number of microbes at the beginning and end of fermentation, measuring pH, and organoleptic observations of the color of fermented cocoa beans. The results of the experiment can be concluded that there is an increase in the addition of mixed inoculum (Saccharomyces cerevisiae and Lactobacillus lactis) affecting the influence of chemical changes of the substrate during fermentation; the temperature value in cocoa fermentation ranges from 30 – 32 oC, the pH value in cocoa fermentation does not have a significant increase; and the color of cocoa beans from mixed inoculum fermentation is close to good with a dominant brown color against purple

Comparative genomic analysis of Periweissella and the characterization of novel motile species.

The genus Periweissella was proposed as a novel genus in the Lactobacillaceae in 2022. However, the phylogenetic relationship between Periweissella and other heterofermentative lactobacilli, and the genetic and physiological properties of this genus remain unclear. This study aimed to determine the phylogenetic relationship between Periweissella and the two closest genera, Weissella and Furfurilactobacillus, by the phylogenetic analysis and calculation of (core gene) pairwise average amino acid identity. Targeted genomic analysis showed that fructose bisphosphate aldolase was only present in the genome of Pw. cryptocerci. Mannitol dehydrogenase was found in genomes of Pw. beninensis, Pw. fabaria, and Pw. fabalis. Untargeted genomic analysis identified the presence of flagellar genes in Periweissella but not in other closely related genera. Phenotypes related to carbohydrate fermentation and motility matched the genotypes. Motility genes were organized in a single operon and the proteins shared a high amino acid similarity in the genus Periweissella. The relatively low similarity of motility operons between Periweissella and other motile lactobacilli indicated the acquisition of motility by the ancestral species. Our findings facilitate the phylogenetic, genetic, and phenotypic understanding of the genus Periweissella.ImportanceThe genus Periweissella is a heterofermentative genus in the Lactobacillaceae which includes predominantly isolates from cocoa fermentations in tropical climates. Despite the relevance of the genus in food fermentations, genetic and physiological properties of the genus are poorly characterized and genome sequences became available only after 2020. This study characterized strains of the genus by functional genomic analysis, and by determination of metabolic and physiological traits. Phylogenetic analysis revealed that Periweissella is the evolutionary link between rod-shaped heterofermentative lactobacilli and the coccoid Leuconostoc clade with the genera Weissella and Furfurilactobacillus as closest relatives. Periweissella is the only heterofermentative genus in the Lactobacillaceae which comprises predominantly motile strains. The genomic, physiological, and metabolic characterization of Periweissella may facilitate the potential use of strains of the genus as starter culture in traditional or novel food fermentations.

Fungal inhibition by lactic acid bacteria (LAB) is modulated in vitro by cocoa fermentation-related conditions: towards a biocontrol of fungi in processing cocoa

Fungal growth impair strongly the quality of processed cocoa, leading to low crops value for farmers and manufacturers. The antifungal activity of lactic acid bacteria (LAB) from fermenting cocoa was analyzed and the conditions of fungal inhibition were evaluated. Fungi were isolated from stored fermented and dried cocoa beans on sabouraud plate medium. The results showed that, LAB strongly inhibit fungi isolated from fermented cocoa beans notably Mucors and Aspergilli, with Lactobacillus plantarum inducing the stronger inhibition whereas Leuconostoc mesenteroides produced a weaker inhibition. Acids production assayed by HPLC was found to be not related to antifungal activity, since LAB strains producing strong antifungal activity were not necessarly the best acid producers. Maximum fungal inhibition occured at 35 °C in Lactobacillus plantarum and Leuconostoc mesenteroides but also at 30 °C in Lactobacllus casei. Likewise, a narrow acidic pH range (3.0-4.0) allowed full expression of fungal inhibtion in LAB analyzed that decreased progressively toward pH 7.0 and failed at alkalin pH. However, sucrose even at high content (8%) was found to have no effect on antifungal activity of LAB, but its bioconversion product glucose and fructose decreased this activity when concentrations were set at 6 et 2 %, respectively. All together, the results suggest that, LAB may effectively exert fungal inhibition in a narrow timing of cocoa fermentation, with pulp contained sugars as metabolic regulators of this inhibition. These results may contribute to a better management of LAB as starter culture for an efficient inhibition of fungal growth and prevention of cocoa contamination from mycotoxins.
 Key Words : Antifungal activity ; lactic acid bacteria ; cocoa fermentation ; inhibition variation; pH and temperature ; sugars content.

An overview: the effect of fermentation and roasting methods on cocoa quality

Cocoa is Indonesia’s most important commodity and one that can contribute to economic development but has a problem, which is the low quality of cocoa produced. Differences in cocoa type, roasting and fermentation affect the quality content of cocoa such as antioxidants, free fatty acid and the fat profile of cocoa. Of the different methods of cocoa bean processing that affect the changes in antioxidant compounds are fermentation and roasting. Fermentation is best with 6 days in producing the aroma flavor of cocoa precursors and bulk cocoa type with an optimal time of 5-7 and fine cocoa type with an optimal time of 3-4 days. Superheated steam method slows down the rate of decline in antioxidant content, this condition is different from the conventional roasting method commonly used because it removes a lot of cocoa antioxidant content and accelerates the decline in antioxidant activity. Non-fermentation treatment results in low free fatty acid values while fermentation treatment can increase free fatty acid content but can be controlled by the addition of alkalization. on the fat profile geographical location such as differences in country of origin affects the content of fatty acid composition but fermentation and roasting processes has no significant effect on the fatty acid composition of cocoa.

Influence of cocoa varieties on carbohydrate composition and enzymatic activity of cocoa pulp

In Brazil, after the witch's broom disease incidence, diverse cocoa hybrids were developed, and variations were reported on their composition and characteristics. Based on this, the present study aimed to evaluate the pulp composition of several cocoa hybrids in order to better understand these variations. Results show that cocoa pulp is composed, on average, of 76 % sugar, and a wide variation (20 %) was observed in sugar content between hybrids. Regarding the sugar profile, a prevalence of reducing sugars was observed. Pod origin also plays an important role in pulp composition, with variations between hybrids from Espírito Santo and Bahia states. In relation to the degree of ripeness, ripe pods showed higher fructose and glucose content, while unripe pods presented mainly sucrose. Similar to sugars, the cello-oligossacharides profile was influenced by the degree of pod ripeness and origin and most ripe samples presented mainly cellobiose, cellotriose and cellotetrose. In addition, the prebiotic potential of cocoa pulp was highlighted by cello-oligossacharides digestion assay which exhibited low rates of degradation. Varying enzymatic activity was observed amongst different pulp hybrids, with polyphenol oxidase showing a higher variation when compared to invertase and polygalacturonase ranging. This study shows that the pod hybrid, origin and ripening degree may change the cocoa pulp composition. Therefore, it is very important to understand and evaluate these variations, in order to obtain better results in pulp utilization either in cocoa fermentation or as a coproduct.

Effects of adding cocoa fermentation medium on cigar leaves in agricultural fermentation stage.

Background and Objective: With the development of the world economy and the integration of cultures, the Chinese cigar market has shown a significant upward trend. However, high-quality cigar leaves are mostly produced in Dominica, Cuba, Nicaragua and other places. In contrast, Chinese cigar leaves have problems such as insufficient aroma, which has become one of the main factors restricting the development of Chinese cigars. Adding medium to ferment is a traditional method in the cigar industry. At present, it mostly relies on manual experience, and lacks systematic and scientific research. At the same time, the addition of medium fermentation is mainly concentrated in the industrial fermentation process, and has not yet begun to be applied in the agricultural fermentation process. In this study, the medium was added to the agricultural fermentation process for the first time to explore the possibility of the application. The effects of adding cocoa medium to ferment on the chemical composition, sensory quality and surface microbial diversity of eggplant core cigar leaves were investigated.wrapper. Method: With Dexue 7' as the experimental material, the changes of main chemical components of wrapper fermented with water and cocoa medium were determined, and microbial community structure on the surface and relative abundance of cigar leaves at different turning periods were analyzed, and the functional genes were predicted. The results of the study were as follows: 1) The results of sensory evaluation showed that the addition of cocoa medium could highlight the aroma of bean, cocoa and coffee, improve the sweetness and fluency and the combustibility of cigar leaves. 2) The addition of cocoa medium increased the contents of proline and malic acid which were positively correlated with sensory quality, and decreased the contents of citric acid, linoleic acid, basic amino acids and aromatic amino acids which were negatively correlated with sensory quality. 3) The addition of cocoa medium increased the total amount of aroma components in cigar leaves, especially carotenoid degradation products, and changed the structural composition of some aroma substances in wrappercigar leaves. 4) The similarity of species composition between the water-added group and the cocoa-added group was higher, but the dominant microorganisms were more concentrated. Staphylococcus and Arthrobacter maintained a high relative abundance throughout the fermentation process, which may be the key microorganisms in the agricultural fermentation stage. 5) The addition of cocoa medium increased the expression abundance of related functional genes in cigar leaves, accelerated the fermentation process of cigar leaves, and bacteria played a major role in the fermentation process. Conclusion: Adding cocoa medium in the agricultural fermentation stage, the changes of bacterial community and dominant flora on the surface of cigar leaves are the main factors affecting their internal chemical components, and the addition of media has a positive effect on tobacco fermentation.

Yeast strains do have an impact on the production of cured cocoa beans, as assessed with Costa Rican Trinitario cocoa fermentation processes and chocolates thereof.

The microbiological and metabolic outcomes of good cocoa fermentation practices can be standardized and influenced through the addition of starter culture mixtures composed of yeast and bacterial strains. The present study performed two spontaneous and 10 starter culture-initiated (SCI) cocoa fermentation processes (CFPs) in Costa Rica with local Trinitario cocoa. The yeast strains Saccharomyces cerevisiae IMDO 050523, Hanseniaspora opuntiae IMDO 020003, and Pichia kudriavzevii IMDO 060005 were used to compose starter culture mixtures in combination with the lactic acid bacterium strain Limosilactobacillus fermentum IMDO 0611222 and the acetic acid bacterium strain Acetobacter pasteurianus IMDO 0506386. The microbial community and metabolite dynamics of the cocoa pulp-bean mass fermentation, the metabolite dynamics of the drying cocoa beans, and the volatile organic compound (VOC) profiles of the chocolate production were assessed. An amplicon sequence variant approach based on full-length 16S rRNA gene sequencing instead of targeting the V4 region led to a highly accurate monitoring of the starter culture strains added, in particular the Liml. fermentum IMDO 0611222 strain. The latter strain always prevailed over the background lactic acid bacteria. A similar approach, based on the internal transcribed spacer (ITS1) region of the fungal rRNA transcribed unit, was used for yeast strain monitoring. The SCI CFPs evolved faster when compared to the spontaneous ones. Moreover, the yeast strains applied did have an impact. The presence of S. cerevisiae IMDO 050523 was necessary for successful fermentation of the cocoa pulp-bean mass, which was characterized by the production of higher alcohols and esters. In contrast, the inoculation of H. opuntiae IMDO 020003 as the sole yeast strain led to underfermentation and a poor VOC profile, mainly due to its low competitiveness. The P. kudriavzevii IMDO 060005 strain tested in the present study did not contribute to a richer VOC profile. Although differences in VOCs could be revealed in the cocoa liquors, no significant effect on the final chocolates could be obtained, mainly due to a great impact of cocoa liquor processing during chocolate-making. Hence, optimization of the starter culture mixture and cocoa liquor processing seem to be of pivotal importance.

Cocoa-associated filamentous fungi for the biocontrol of aflatoxigenic Aspergillus flavus.

Aflatoxin and other mycotoxin contamination are major threats to global food security and present an urgent need to secure the global food crop against spoilage by mycotoxigenic fungi. Cocoa material is noted for naturally low aflatoxin contamination. This study was designed to assess the potential for harnessing cocoa-associated filamentous fungi for the biocontrol of aflatoxigenic Aspergillus flavus. The candidate fungi were isolated from fermented cocoa beans collected from four cocoa-growing areas in Ghana. Molecular characterization included Internal Transcribed Spacer (ITS)-sequencing for identification and polymer chain reaction (PCR) to determine mating type. Effects of the candidate isolates on growth and aflatoxin-production by an aflatoxigenic A. flavus isolate (BANGA1) were assessed. Aflatoxin production was monitored by UV fluorescence and quantified by enzyme-linked immunosorbent assay (ELISA). Thirty-six filamentous fungi were cultured and identified as Aspergillus, Cladosporium, Lichtheimia, or Trichoderma spp. isolates. The isolates generally interacted negatively with BANGA1 growth and aflatoxin production. The Aspergillus niger and Aspergillus aculeatus biocontrol candidates showed the strongest colony antagonism (54%-94%) and reduction in aflatoxin production (12%-50%) on agar. In broth, the A. niger isolates reduced aflatoxin production by up to 97%. Metabolites from the A. niger isolates showed the strongest inhibition of growth by BANGA1 and inhibited aflatoxin production. Four of the candidate isolates belonged to the MAT1-1 mating type and 12 identified as MAT1-2. This may be indicative of the potential for genetic recombination events between fungi in the field, a finding which is particularly relevant to the risk posed by A. flavus biocontrol measures that rely on atoxigenic A. flavus strains.

Physicochemical and Sensory Analysis of Cocoa-Coffee Blend Drink Using the Check-All-That-Apply (CATA)

Chocolate drinks are widely consumed due to their rich flavor and polyphenol content. Previous research has highlighted the flavor richness and high polyphenol content of Arabica coffee. This study aims to examine the effects of adding Arabica coffee on the physical characteristics and sensory profiles of chocolate drinks. The composition of the chocolate drinks used in this study includes fermented cocoa powder, Java Preanger Arabica coffee powder, Cascara Arabica, and stevia sweetener. Physicochemical analysis methods encompass color testing, pH measurement, and total soluble solids (TSS) determination. The five tested formulations resulted in a slightly dark yellowish-red color, with pH values ranging from 5.82 ± 0.10 to 6.22 ± 0.08, and TSS content ranging from 5.17 ± 0.76 to 5.90 ± 0.46. In addition to physicochemical analysis, sensory analysis was conducted using the Check-All-That-Apply (CATA) method facilitated by XLSTAT. Although no formulations matched the ideal product, the most preferred formulation consisted of 80% cocoa and 20% coffee, attributed to its sweet flavor. With lower percentages of coffee addition, attributes such as chocolate flavor, sweetness, caramel flavor, and milk flavor became more prominent in the chocolate drinks.

Effect of the maturity stage, genotype, and geographical location on the physicochemical characteristics of the cocoa bean during fermentation

An essential aspect of competing in the world market is the quality of the cocoa bean, which is influenced by different factors ranging from genetics to post-harvest processes. The aim of this research was to determine the effect of the maturity stage (2, 3, and 4), genotype (CCN51, ICS95, and TSH565), and geographical location (Algeciras and Garzón, Colombia) of cocoa on the pH and titratable acidity of the pulp and bean, and the fermentation index of the bean during days 0, 3 and 7 of fermentation. The response variables were analyzed through an analysis of variance (ANOVA), and multiple comparisons (pairwise) were made using Tukey's test with a significance level of α = 0.05. Additionally, a longitudinal analysis was carried out as there are measurements over time, and autocorrelation of individuals over time can occur, allowing contrasting the results through another analysis. The results showed that the factor with the strongest influence on the physicochemical characteristics evaluated was the genotype, affecting the bean and pulp pH and the fermentation index. Likewise, it was possible to show that, of the three genotypes evaluated, CCN51 stood out for presenting less bean acidity and higher bean pH and fermentation index. The genotype is the factor with the strongest effect during cocoa fermentation. This solves bean quality heterogeneity, the main international marketing problem.