The cacao fruit is a rich source of polyphenols, including flavonoids and phenolic acids, which possess significant health benefits. The accurate identification and quantification of these bioactive compounds extracted from different parts of the cacao fruit, such as pods, beans, nibs, and cacao shells, require specific treatment conditions and analytical techniques. This review presents a comprehensive comparison of extraction processes and analytical techniques used to identify and quantify polyphenols from various parts of the cacao fruit. Additionally, it highlights the environmental impact of these methods, exploring the challenges and opportunities in selecting and utilizing extraction, analytical, and impact assessment techniques, while considering polyphenols\' yield. The review aims to provide a thorough overview of the current knowledge that can guide future decisions for those seeking to obtain polyphenols from different parts of the cacao fruit.

The food industry is increasingly striving to produce probiotics-based food and beverages using sustainable processes. Therefore, the use of by-products in product development has been investigated by several authors. The aim of this work was to investigate the effects of cocoa bean shell infusion in the production of kombucha through microbiological and genetic characterization. Three beverage formulations were prepared, one based on black tea (KBT), one based on cocoa bean shell infusion (KCS) and one containing 50 % black tea and 50 % cocoa shell infusion (KBL). The infusions were prepared with water, filtered, and sucrose was added. They were then homogenized and a portion of finished kombucha and SCOBY (symbiotic culture of bacteria and yeast) were added. Fermentation took place for 13 days and aliquots were collected every three days for physicochemical and microbial count analyses. Samples from the last day of fermentation were sent for DNA sequencing, extraction and quantification. The results were subjected to analysis of variance and compared by using Tukey\'s test (p < 0.05). The results show that there was a significant decrease in pH over time in all samples, while the titratable acidity increased, indicating an acidification of the beverage due to the production of organic acids. There was an increase in lactic acid bacterial colonies in all the formulations, which have a probiotic nature and are not always found in this type of beverage. Regarding the taxonomic classification of the samples, microorganisms of the kingdoms Fungi and Bacteria, of the families Saccharomycetaceae and Acetobacteraceae, were found in KBT, KCS and KBL, but with different microbiological compositions, with different amounts of yeasts and bacteria. Therefore, the use of by-products such as cocoa bean shell in the production of kombucha can contribute to the reduction of waste in the food industry and, at the same time, accelerate fermentation increasing the presence of lactic acid bacteria when compared to black tea.

Cocoa tree plantations aim to harvest grains found in the cob to produce cocoa and chocolate. There has been a growing interest in valorizing the secondary components of the cocoa fruit, such as the peel, placenta, and mucilage/pulp, as valuable sources of nutrients for healthy food preparation. In other words, by-products derived from these raw materials are an exploitable source of nutrients in the preparation of healthy food. In the present study, two varieties of cocoa, National Cocoa Fino de Aroma (NCFA) and Colección Castro Naranjal 51 (CCN-51), were evaluated and harvested during both dry and rainy seasons. This evaluation was based on the profiling of the cob, peel, grain, placenta, and mucilage in different stages of ripeness (underripe, ripe, and overripe). Also, from the ripe raw material, a fermented beverage prototype was developed, such as kombucha, with different concentrations of mucilage (40, 60, 80, and 100 g/L). Physicochemical analyses, such as acidity, °Brix, pH, moisture, ash, protein, fat, fiber, vitamins, sugars, and polyphenols of the raw mucilage material and acidity, °Brix, and pH values of the fermented kombucha, were carried out. The best performances were obtained with the CCN-51 variety in the rainy season. Among the fermented drink panelists, the CN40 treatment (Nacional Mucilage + 40 g/L of sugar) received the highest acceptability and was considered the best. Given its efficiency, nutritional content, and potential applications, this product presents a promising strategy to address Sustainable Development Goals related to zero hunger, health and well-being, and climate action.

Similar to cacao pod rot, cherelle wilt decreases production from cacao fields. Among all known fungal pathogens of the cacao, Colletotrichum spp. are common infectious agents that affect the cherelles and pods of cacao; thus, cacao diseases are often classified by stage. Therefore, knowing whether these pathogens are common in both fruit stages is necessary for implementing disease control measures. Symptoms of cherelle wilt were found in cacao plants in Pangasinan, Philippines, in 2022. The fungal strain obtained from the lesion was found to be pathogenic towards cherelles, but not towards pods. The strain was classified as an unknown species belonging to the gigasporum species complex, based on the morphological and molecular phylogenetic analyses of ITS, GAPDH, CHS1, ACT, and TUB2. We propose Colletotrichum kapreanum sp. nov. as a causal agent of cacao cherelle wilt, but not pod rot.

Chagas disease (CHD) is the highest economic burden parasitosis worldwide and the most important cardiac infection, without therapeutic alternatives to halt or reverse its progression. In CHD-experimental models, antioxidant and anti-inflammatory compounds have demonstrated therapeutic potential in cardiac dysfunction. Theobroma cacao polyphenols are potent natural antioxidants with cardioprotective and anti-inflammatory action, which are susceptible to degradation, requiring technological approaches to guarantee their protection, stability, and controlled release. Here, 21 cocoa polyphenol-rich microencapsulates were produced by spray-drying and freeze-drying techniques using two wall materials (maltodextrin and gum arabic). Chemical (total and individual phenolic content and antioxidant activity), structural (morphology), and biological parameters (cytotoxicity, trypanocidal, antioxidant, and immunomodulatory activities) were assessed to determine the most efficient microencapsulation conditions on Trypanosoma cruzi-infected myocardioblast and macrophage cells. Significant antiproliferative properties against infected cells (superior to benznidazole) were found in two microencapsulates which also exhibited cardioprotective properties against oxidative stress, inflammation, and cell death.

The aim of this study was to determine the chemical structure and biological activity of melanoidin fractions derived from cocoa beans, carob kibbles, and acorns roasted at different temperature-time conditions. The results showed that plant origin and roasting conditions had significant effects on the chemical composition, structural features, and morphology of melanoidins. All tested melanoidins exhibited significant antioxidant properties in three in vitro assays. In addition, they show significant in vitro anti-inflammatory activity by reducing lipoxygenase. The results from MTT assay showed that the all studied melanoidins had a cytotoxic effect against SW-480 cells in a dose- and time-dependent manner. Furthermore, the most pronounced activity was observed for acorn melanoidins. This is a unique finding, as the specific cytotoxic effect has not been reported for cocoa, carob and acorn melanoidins, and opens up a great opportunity to develop a potential novel cytotoxic agent against deadly colon cancer in the future.

The present work was conducted to evaluate the volatile profile of Ecuadorian Forastero, CCN-51, ETT103 and LR14 cocoa beans during traditional fermentation in laurel wood boxes followed by a sun-drying process. Fifty-six volatiles were identified with HS-SPME-GC-MS. Aldehydes, alcohols and ketones were the compounds that mainly characterized the fresh cocoa. The main compounds formed during the anaerobic fermentation step were esters and acids, while in the aerobic fermentation step, an increase in ester-, aldehyde- and acid-type compounds was observed. Finally, after the drying step, a notable increase in the acid (i.e., acetic acid) content was the predominant trend. According to the genotypes, ETT103 presented high contents of terpenes, alcohols, aldehydes and ketones and low contents of unfavorable acid compounds. The CCN-51 and LR14 (Trinitarian) varieties stood out for their highest amounts in acids (i.e., acetic acid) at the end of primary processing. Finally, the Forastero cocoa beans were highlighted for their low acid and high trimethylpyrazine contents. According to the chemometric and Venn diagram analyses, ETT-103 was an interestingly high-aromatic-quality variety for cocoa gourmet preparations. The results also showed the need for good control of the processing steps (using prefermentative treatments, starter cultures, etc.) on Ecuadorian genotypes of Trinitarian origin.

Probiotics are known for their health-promoting properties and are recognized as beneficial microorganisms. The current investigation delves into the isolation and comprehensive in vitro characterization of lactic acid bacteria (LAB) obtained from the Indian-origin Theobroma cacao L. Forastero variety to assess their potential as probiotic candidates. Eleven LAB isolates were obtained, and among them, five exhibited classical LAB traits. These five isolates underwent rigorous in vitro characterization to evaluate their suitability as probiotics. The assessments included resilience against acid and bile salts, which are crucial for probiotic viability. Additionally, the isolates were subjected to simulated gastric and pancreatic fluids and lysozyme exposure to assess their survival rates. Auto- aggregation, co-aggregation, hydrophobicity, and exopolysaccharide production were also examined. The inhibitory potential of α-glucosidase, an enzyme related to glucose metabolism, was measured, and antioxidant activity was evaluated using DPPH and ABTS assays. A safety assessment was conducted to confirm the non-pathogenic nature of the isolates. Among the five isolates, CR2 emerged as a standout candidate with maximal bile salt hydrolase activity, phenol resistance, and lysozyme resistance. CR2 and CYF3 exhibited notable survival rates under simulated conditions. The isolates displayed variable degrees of auto-aggregation, co-aggregation, and hydrophobicity. CR2 exhibited the highest exopolysaccharide production (0.66 mg/mL), suggesting diverse applications in the food industry. CR2 also demonstrated the highest inhibition rate against α-glucosidase (56.55%) and substantial antioxidant activity (79.62% DPPH, 83.45% ABTS). Safety assessment confirmed the non- pathogenic nature of the isolates. Molecular characterization identified CR2 as Lactococcus lactis subsp. lactis and CYF3 as Limnosilactobacillus fermentum. Both strains exhibited commendable probiotic and technological attributes, positioning them as promising candidates for functional foods and beyond. This study provides valuable insights into the in vitro characterization of LAB isolated from Indian Theobroma cacao L., highlighting their potential as probiotic candidates with advantageous traits, including survival in hostile conditions, beneficial enzymatic activities, bioactivity, and other essential attributes.

The roasting process can modulate sensory and physicochemical characteristics of cocoa. This study compared the chemical characteristics of cocoa nibs roasted by a convective oven [slow roasting─(SR)] vs cocoa nibs roasted in a fluidized bed roaster [fast roasting─(FR)] at two temperatures (120 and 140 °C). The contents of sugars, free amino acids (FAAs), polyphenols, acrylamide, 5-hydroxymethylfurfural, and melanoidins were monitored. Roasting reduced fructose, glucose, and sucrose contents by 95, 70, and 55%, respectively. The concentration of total FAAs was reduced up to 40% at 140 °C. The FAA profile revealed that FR favored the reactivity of some amino acids (Leu, Lys, Phe, and Val) relevant in the formation of aroma compounds and melanoidins. FR resulted in the generation of more intense brown melanoidins, a significant increase in catechin content, a higher formation of acrylamide, and a lower formation of 5-hydroxymethylfurfural in cocoa compared to SR.

Aiming to improve the quality of cocoa, preconditioning of cocoa after harvesting and before fermentation has become an on-farm processing step of great interest in recent times. The present work aimed to evaluate the influence of a pre-drying process on the volatile composition of Ecuadorian bulk (Forastero and CCN-51) and fine-flavour (ETT103 and LR14) cocoa at the end of primary processing. A total of 63 volatile compounds including aldehydes, alcohols, acids, ketones, esters, terpenes, lactones and other miscellaneous compounds were identified in cocoa samples by HS-SPME-GC-MS. The use of a pre-drying step revealed a varietal homogenization and a reduction in the fermentation time, making this preconditioning step an inexpensive and attractive option for farmers. Moreover, different varietal behaviour was observed after pre-drying, the fine-flavour varieties obtaining a clear improvement in aromatic quality with higher levels of compounds imparting positive notes.