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Bibliografia_____________________________________________________________________

(1) Todorovic V, Milenkovic M, Vidovic B, Todorovic Z, Sobajic S. Correlation between Antimicrobial, Antioxidant Activity, and Polyphenols of Alkalized/Nonalkalized Cocoa Powders. J Food Sci. 2017 Apr;82(4):1020-1027. doi: 10.1111/1750-3841.13672. Epub 2017 Mar 8. PMID: 28272800.

Abstract. Many factors can influence antioxidative and antimicrobial characteristics of plant materials. The quality of cocoa as functional food ingredient is influenced through its processing. The main aim of this study was to test if there is difference in polyphenol content, antioxidant capacity, and antimicrobial activity between nonalkalized and alkalized cocoa powders. To estimate polyphenol and flavonoid content in cocoa samples the spectrophotometric microassays were used. Flavan-3ols were determined with reversed-phase high-performance liquid chromatography (RP-HPLC). Antimicrobial activity against 3 Gram positive bacteria, 4 Gram negative bacteria and 1 strain of yeast was determined using broth microdilution method. Total polyphenol content was 1.8 times lower in alkalized cocoa samples than in natural ones. Epicatechin/catechin ratio was changed due to the process of alkalization in favor of catechin (2.21 in natural and 1.45 in alkalized cocoa powders). Combined results of 3 antioxidative tests (DPPH, FRAP, ABTS) were used for calculation of RACI (Relative Antioxidant Capacity Index) and GAS (Global Antioxidant Score) values that were consistently higher in natural than in alkalized cocoa extracts. Obtained results have shown significant correlations between these values and phenolic content (0.929 ≤ r ≤ 0.957, P < 0.01). Antimicrobial activity varied from 5.0 to 25.0 mg/ml (MICs), while Candida albicans was the most sensitive tested microorganism. Cocoa powders subjected to alkalization had significantly reduced content of total and specific phenolic compounds and reduced antioxidant capacity (P < 0.05), but their antimicrobial activity was equal for Gram-positive bacteria or even significantly enhanced for Gram-negative bacteria. © 2017 Institute of Food Technologists®.

Alasti FM, Asefi N, Maleki R, SeiiedlouHeris SS. The influence of three different types and dosage of alkaline on the inherent properties in cocoa powder. J Food Sci Technol. 2020 Jul;57(7):2561-2571. doi: 10.1007/s13197-020-04293-w. 

Montagna MT, Diella G, Triggiano F, Caponio GR, De Giglio O, Caggiano G, Di Ciaula A, Portincasa P. Chocolate, "Food of the Gods": History, Science, and Human Health. Int J Environ Res Public Health. 2019 Dec 6;16(24):4960. doi: 10.3390/ijerph16244960. 

Abstract. Chocolate is well known for its fine flavor, and its history began in ancient times, when the Maya considered chocolate (a cocoa drink prepared with hot water) the "Food of the Gods". The food industry produces many different types of chocolate: in recent years, dark chocolate, in particular, has gained great popularity. Interest in chocolate has grown, owing to its physiological and potential health effects, such as regulation of blood pressure, insulin levels, vascular functions, oxidation processes, prebiotic effects, glucose homeostasis, and lipid metabolism. However, further translational and epidemiologic studies are needed to confirm available results and to evaluate other possible effects related to the consumption of cocoa and chocolate, verifying in humans the effects hitherto demonstrated only in vitro, and suggesting how best to consume (in terms of dose, mode, and time) chocolate in the daily diet.

Ferri C, Desideri G, Ferri L, Proietti I, Di Agostino S, Martella L, Mai F, Di Giosia P, Grassi D. Cocoa, blood pressure, and cardiovascular health. J Agric Food Chem. 2015 Nov 18;63(45):9901-9. doi: 10.1021/acs.jafc.5b01064. 

Abstract. High blood pressure is an important risk factor for cardiovascular disease and cardiovascular events worldwide. Clinical and epidemiological studies suggest that cocoa-rich products reduce the risk of cardiovascular disease. According to this, cocoa has a high content in polyphenols, especially flavanols. Flavanols have been described to exert favorable effects on endothelium-derived vasodilation via the stimulation of nitric oxide-synthase, the increased availability of l-arginine, and the decreased degradation of NO. Cocoa may also have a beneficial effect by protecting against oxidative stress alterations and via decreased platelet aggregation, decreased lipid oxidation, and insulin resistance. These effects are associated with a decrease of blood pressure and a favorable trend toward a reduction in cardiovascular events and strokes. Previous meta-analyses have shown that cocoa-rich foods may reduce blood pressure. Long-term trials investigating the effect of cocoa products are needed to determine whether or not blood pressure is reduced on a chronic basis by daily ingestion of cocoa. Furthermore, long-term trials investigating the effect of cocoa on clinical outcomes are also needed to assess whether cocoa has an effect on cardiovascular events. A 3 mmHg systolic blood pressure reduction has been estimated to decrease the risk of cardiovascular and all-cause mortality. This paper summarizes new findings concerning cocoa effects on blood pressure and cardiovascular health, focusing on putative mechanisms of action and "nutraceutical " viewpoints.

Jean-Marie E, Jiang W, Bereau D, Robinson JC. Theobroma cacao and Theobroma grandiflorum: Botany, Composition and Pharmacological Activities of Pods and Seeds. Foods. 2022 Dec 8;11(24):3966. doi: 10.3390/foods11243966. 

Abstract. Cocoa and cupuassu are evergreen Amazonian trees belonging to the genus Theobroma, with morphologically distinct fruits, including pods and beans. These beans are generally used for agri-food and cosmetics and have high fat and carbohydrates contents. The beans also contain interesting bioactive compounds, among which are polyphenols and methylxanthines thought to be responsible for various health benefits such as protective abilities against cardiovascular and neurodegenerative disorders and other metabolic disorders such as obesity and diabetes. Although these pods represent 50-80% of the whole fruit and provide a rich source of proteins, they are regularly eliminated during the cocoa and cupuassu transformation process. The purpose of this work is to provide an overview of recent research on cocoa and cupuassu pods and beans, with emphasis on their chemical composition, bioavailability, and pharmacological properties. According to the literature, pods and beans from cocoa and cupuassu are promising ecological and healthy resources.

Tan TYC, Lim XY, Yeo JHH, Lee SWH, Lai NM. The Health Effects of Chocolate and Cocoa: A Systematic Review. Nutrients. 2021 Aug 24;13(9):2909. doi: 10.3390/nu13092909. 

Abstract. Chocolate has a history of human consumption tracing back to 400 AD and is rich in polyphenols such as catechins, anthocyanidins, and pro anthocyanidins. As chocolate and cocoa product consumption, along with interest in them as functional foods, increases worldwide, there is a need to systematically and critically appraise the available clinical evidence on their health effects. A systematic search was conducted on electronic databases such as MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials (CENTRAL) using a search strategy and keywords. Among the many health effects assessed on several outcomes (including skin, cardiovascular, anthropometric, cognitive, and quality of life), we found that compared to controls, chocolate or cocoa product consumption significantly improved lipid profiles (triglycerides), while the effects of chocolate on all other outcome parameters were not significantly different. In conclusion, low-to-moderate-quality evidence with short duration of research (majority 4-6 weeks) showed no significant difference between the effects of chocolate and control groups on parameters related to skin, blood pressure, lipid profile, cognitive function, anthropometry, blood glucose, and quality of life regardless of form, dose, and duration among healthy individuals. It was generally well accepted by study subjects, with gastrointestinal disturbances and unpalatability being the most reported concerns.

Schwan RF, Wheals AE. The microbiology of cocoa fermentation and its role in chocolate quality. Crit Rev Food Sci Nutr. 2004;44(4):205-21. doi: 10.1080/10408690490464104. 

Abstract. The first stage of chocolate production consists of a natural, seven-day microbial fermentation of the pectinaceous pulp surrounding beans of the tree Theobroma cacao. There is a microbial succession of a wide range of yeasts, lactic-acid, and acetic-acid bacteria during which high temperatures of up to 50 degrees C and microbial products, such as ethanol, lactic acid, and acetic acid, kill the beans and cause production of flavor precursors. Over-fermentation leads to a rise in bacilli and filamentous fungi that can cause off-flavors. The physiological roles of the predominant micro-organisms are now reasonably well understood and the crucial importance of a well-ordered microbial succession in cocoa aroma has been established. It has been possible to use a synthetic microbial cocktail inoculum of just 5 species, including members of the 3 principal groups, to mimic the natural fermentation process and yield good quality chocolate. Reduction of the amount of pectin by physical or mechanical means can also lead to an improved fermentation in reduced time and the juice can be used as a high-value byproduct. To improve the quality of the processed beans, more research is needed on pectinase production by yeasts, better depulping, fermenter design, and the use of starter cultures.

Cárdenas A, Mojica L, Coronado-Cáceres L, Castillo-Herrera GA. Unlocking the Alkaloid Biological Potential of Chili Pepper (Capsicum spp.), Cacao (Theobroma cacao L.), and Coffee (Coffea spp.) Byproducts: Characterization, Non-Conventional Extraction, Applications, and Future Perspectives. Molecules. 2025 Sep 18;30(18):3795. doi: 10.3390/molecules30183795. 

Abstract. Chili peppers (Capsicum spp.), cacao (Theobroma cacao L.), and coffee (Coffea spp.) are important crops worldwide. Nearly 35%, 80%, and 45% of the respective fruits are underutilized or discarded, representing a considerable economic loss. This work reviews and analyzes the environmental factors that influence the concentration of the main alkaloids in these crops, including capsaicin, theobromine, and caffeine. Their reported anti-inflammatory, cardioprotective, neuroprotective, and cytotoxic properties are also reviewed. This work explores strategies for the revalorization of these crops, comparing alkaloid extraction methods that use non-conventional techniques, including supercritical fluid extraction (SFE), ultrasound-assisted extraction (UAE), high-pressure and -temperature extraction (HPTE), pressurized liquid extraction (PLE), pressurized hot water extraction (PHWE), enzyme-assisted extraction (EAE), and pulsed electric field-assisted extraction (PEFAE), and their combination to enhance the recovery of capsaicin, theobromine, and caffeine, leading to sustainable and innovative uses of these crops' byproducts. Capsaicin, theobromine, and caffeine alkaloids are promising ingredients for the development of functional foods, cosmeceuticals, and pharmaceuticals.

Díaz-Valderrama JR, Leiva-Espinoza ST, Aime MC. The History of Cacao and Its Diseases in the Americas. Phytopathology. 2020 Oct;110(10):1604-1619. doi: 10.1094/PHYTO-05-20-0178-RVW. 

Abstract. Cacao is a commodity crop from the tropics cultivated by about 6 million smallholder farmers. The tree, Theobroma cacao, originated in the Upper Amazon where it was domesticated ca. 5450 to 5300 B.P. From this center of origin, cacao was dispersed and cultivated in Mesoamerica as early as 3800 to 3000 B.P. After the European conquest of the Americas (the 1500s), cacao cultivation intensified in several loci, primarily Mesoamerica, Trinidad, Venezuela, and Ecuador. It was during the colonial period that cacao diseases began emerging as threats to production. One early example is the collapse of the cacao industry in Trinidad in the 1720s, attributed to an unknown disease referred to as the "blast". Trinidad would resurface as a production center due to the discovery of the Trinitario genetic group, which is still widely used in breeding programs around the world. However, a resurgence of diseases like frosty pod rot during the republican period (the late 1800s and early 1900s) had profound impacts on other centers of Latin American production, especially in Venezuela and Ecuador, shifting the focus of cacao production southward, to Bahia, Brazil. Production in Bahia was, in turn, dramatically curtailed by the introduction of witches' broom disease in the late 1980s. Today, most of the world's cacao production occurs in West Africa and parts of Asia, where the primary Latin American diseases have not yet spread. In this review, we discuss the history of cacao cultivation in the Americas and how that history has been shaped by the emergence of diseases.

Martin MÁ, Ramos S. Impact of cocoa flavanols on human health. Food Chem Toxicol. 2021 May;151:112121. doi: 10.1016/j.fct.2021.112121. Epub 2021 Mar 13. PMID: 33722594.

Kerimi A, Williamson G. The cardiovascular benefits of dark chocolate. Vascul Pharmacol. 2015 Aug;71:11-5. doi: 10.1016/j.vph.2015.05.011. Epub 2015 May 27. PMID: 26026398.