New light on the use of Theobroma cacao by Late Classic Maya.

Cacao seeds, Theobroma cacao, provide the basis for a ceremonially important Mesoamerican food. Past efforts to identify cacao in ceramics focused on highly decorative vessel forms associated with elite ceremonial contexts, creating assumptions as to how cacao was distributed and who could access it. This study examines 54 archaeological ceramic sherds from El Pilar (Belize/Guatemala) of Late Classic (600 to 900 CE) residential and civic contexts representing a cross-section of ancient Maya inhabitants. Identification of cacao in ancient sherds has depended on the general presence of theobromine; we used the discrete presence of theophylline, a unique key biomarker for cacao in the region. Analysis was done by grinding off all outside surfaces to reduce contamination, pulverizing the inner clay matrix, extracting absorbed molecules, and concentrating the extractions. In order to obtain especially high selectivity and low limits of detection, our study utilized the technique of resonance-enhanced multiphoton ionization coupled with laser-desorption jet-cooling mass spectrometry. This technique isolates molecules in the cold gas phase where they can be selectively ionized through a resonant two-photon process. Of the sherds analyzed, 30 samples (56%) were found to contain significant amounts of theophylline and thus test positive for cacao. Importantly, cacao is present in all contexts, common to all Maya residents near and far from centers.

Desenvolvimento de achocolatados em pó com adição de subprodutos de frutas, processados por spray dryer e com modificador reológico / Development of powdered chocolate added with fruit byproducts, processed by spray dryer and rheological modifier

O achocolatado em pó é um dos derivados do cacau com maior inserção econômica e cultural em diversos países. O objetivo deste estudo foi avaliar a adição de ingredientes diferenciados nesse tipo de produto, como modificadores reológicos e fruta, bem como à alteração no tipo de cacau utilizado, ocasionando mudanças sensoriais e nutricionais positivas ao produto. O fruto sugerido neste estudo foi o cupuaçu (Theoboroma grandiflorum), fruto típico da região Norte, que apresenta excelente qualidade nutricional. Foram desenvolvidas 7 formulações de achocolatado por método convencional após simples mistura (padrão, com cacau alcalino, com cacau orgânico, com polpa de cupuaçu, com amido pré-gelatinizado, com amido pré-gelatinizado + polpa de cupuaçu, com goma guar, com goma guar + polpa de cupuaçu) e 4 formulações processadas por spray dryer após a simples mistura (padrão, com polpa de cupuaçu, com amido pré-gelatinizado, com amido + polpa de cupuaçu). Todas as formulações foram avaliadas quanto à composição nutricional, calorimetria exploratória de varredura (DSC), análises físico-químicas, reológica, quantificação dos compostos fenólicos e avaliação da capacidade antioxidante por métodos in vitro. Em seguida foi realizada análise sensorial com as formulações: padrão, com polpa de cupuaçu e com amido + polpa de cupuaçu. O achocolatado padrão apresentou tempo de mistura de 38 min, o que foi utilizado como parâmetro para as demais formulações. Os achocolatados que continham polpa de cupuaçu apresentaram maior teor proteico (14,5 a 16,3 g/100g) quando comparados com o padrão (13,6 g/100g). Todos os achocolatados apresentaram umidade entre 1,2% e 3,7%, e atividade de água entre 0,13 e 0,57, considerados microbiologicamente estáveis, sendo bom para a vida útil do produto. Os achocolatados obtiveram tempo de molhabilidade entre 07:15 min e 15:06 min; solubilidade de 1,56 IR% a 7,44 IR%; tamanho de partícula variando entre 0,216 mm e 0,347 mm (partículas finas). O uso do spray dryer não teve impacto significativo nas características físicas das formulações, assim como a utilização dos diferentes tipos de cacau não afetou a composição nutricional e qualidade física dos achocolatados. Houve aumento (p< 0,05) para o tempo de molhabilidade e solubilidade do achocolatado com cacau orgânico em comparação com o padrão (13:30 e 9:33 min; 2,64 e 1,56 IR%, respectivamente). A transição vítrea variou entre 35,2 a 35,7 mW enquanto o ponto de carbonização ficou entre 237,4 a 243,6 mW, indicando que a adição dos agentes espessantes e/ou do cupuaçu não interferiu (p<0,05) na análise térmica dos achocolatados. Todos os achocolatados diluídos em leite apresentaram-se como pseudoplásticos, com aumento de viscosidade nas menores temperaturas, conforme esperado. O achocolatado com cacau orgânico apresentou o maior teor de compostos fenólicos (8,27 mg AG g-1) enquanto observou-se redução no conteúdo de fenólicos nos produtos processados por spray dryer. Os achocolatados apresentaram capacidade antioxidante entre 31,76 µMETrolox/g e 75,62 µMETrolox/g, pelo método do DPPH. A adição do cupuaçu levou ao aumento da capacidade de sequestro de radicais DPPH quando comprados com o padrão (p<0,05). Não foi observada diferença significativa pelo método FRAP. A avaliação sensorial obteve aceitação situada na região positiva da escala (5 a 7). Os achocolatados formulados apresentam formulações adequadas a sua comercialização, com agregação de valor nutricional e econômico

The chocolate powdered is a cocoa-derived with greater economic and cultural integration in several countries. The objective of this study was to evaluate the addition of different ingredients in this type of product, such as rheological modifiers and fruit, as well as the change in the type of cocoa used, causing positive sensory and nutritional changes to the product. The fruit suggested in this study was cupuassu (Theoboroma grandiflorum), a typical fruit from the northern region, which has excellent nutritional quality. Seven powdered chocolate formulations were developed by conventional method after simple mixing (standard, with alkaline cocoa, with organic cocoa, with cupuassu pulp, pre-gelatinized starch, pre-gelatinized starch + cupuassu pulp, guar gum, with guar gum + cupuassu pulp) and 4 formulations processed by spray dryer after simple mixing (standard, with cupuassu pulp, pre-gelatinized starch, starch + cupuassu pulp). All formulations were evaluated for nutritional composition, differential scanning calorimetry (DSC), physicochemical, rheological analyzes, quantification of phenolic compounds and antioxidant capacity evaluation by in vitro methods. Then, sensory analysis was performed with the formulations: standard, with cupuassu pulp and starch + cupuassu pulp. The standard powdered chocolate had a mixing time of 38 min, which was used as parameter for the other formulations. The powdered chocolate containing cupuassu pulp had higher protein content (14.5 to 16.3 g / 100g) when compared to the standard (13.6 g / 100g). All powdered chocolate presented humidity between 1.2% and 3.7%, and water activity between 0.13 and 0.57, considered microbiologically stable, wich is good for the shelf life of the product. The powdered chocolate obtained wettability time between 07:15 min and 15:06 min; solubility from 1.56 IR% to 7.44 IR%; particle size ranging from 0.216 mm to 0.347 mm (fine particles). The use of the spray dryer had no significant impact on the physical characteristics of the formulations, as well as the use of different types of cocoa did not affect the nutritional composition and physical quality of the powdered chocolate. There was an increase (p <0.05) for the time of wettability and solubility in chocolate powdered formulated with organic cocoa when compared to the standard (9:33 and 13:30 min; IR 2.64 and 1.56%, respectively). The glass transition ranged from 35.2 to 35.7 mW while the carbonization point ranged from 237.4 to 243.6 mW, indicating that the addition of thickening agents and / or cupuassu did not interfere (p <0.05) in the thermal analysis of powdered chocolate. All powdered chocolate when diluted in milk presented as pseudoplastics, with viscosity increase at lower temperatures, as expected. Chocolate powdered with organic cocoa presented the highest content of phenolic compounds (8.27 mg AG g-1) whereas there was a reduction in phenolic content in products processed by spray dryer. The powdered chocolates presented antioxidant capacity between 31.76 µMETrolox / g and 75.62 µMETrolox / g, by the DPPH method. The addition of cupuassu led to increased ability to sequester DPPH radicals when compared to the standard (p <0.05). No significant difference was observed by the FRAP method. Sensory evaluation was accepted in the positive region of the scale (5 to 7). The formulated powdered chocolates have appropriate formulations for marketing, with added nutritional and economic value

Chemical modification of the cocoa shell surface using diazonium salts.

The outer portion of the cocoa bean, also known as cocoa husk or cocoa shell (CS), is an agrowaste material from the cocoa industry. Even though raw CS is used as food additive, garden mulch, and soil conditioner or even burnt for fuel, this biomass material has hardly ever been investigated for further modification. This article proposes a strategy of chemical modification of cocoa shell to add value to this natural material. The study investigates the grafting of aryl diazonium salt on cocoa shell. Different diazonium salts were grafted on the shell surface and characterized by infrared spectroscopy and scanning electronic microscopy imaging. Strategies were developed to demonstrate the spontaneous grafting of aryl diazonium salt on cocoa shell and to elucidate that lignin is mainly involved in immobilizing the phenyl layer.

Trichoderma species form endophytic associations within Theobroma cacao trichomes.

Trichoderma species are usually considered soil organisms that colonize plant roots, sometimes forming a symbiotic relationship. Recent studies demonstrate that Trichoderma species are also capable of colonizing the above ground tissues of Theobroma cacao (cacao) in what has been characterized as an endophytic relationship. Trichoderma species can be re-isolated from surface sterilized cacao stem tissue, including the bark and xylem, the apical meristem, and to a lesser degree from leaves. SEM analysis of cacao stems colonized by strains of four Trichoderma species (Trichoderma ovalisporum-DIS 70a, Trichoderma hamatum-DIS 219b, Trichoderma koningiopsis-DIS 172ai, or Trichoderma harzianum-DIS 219f) showed a preference for surface colonization of glandular trichomes versus non-glandular trichomes. The Trichoderma strains colonized the glandular trichome tips and formed swellings resembling appresoria. Hyphae were observed emerging from the glandular trichomes on surface sterilized stems from cacao seedlings that had been inoculated with each of the four Trichoderma strains. Fungal hyphae were observed under the microscope emerging from the trichomes as soon as 6h after their isolation from surface sterilized cacao seedling stems. Hyphae were also observed, in some cases, emerging from stalk cells opposite the trichome head. Repeated single trichome/hyphae isolations verified that the emerging hyphae were the Trichoderma strains with which the cacao seedlings had been inoculated. Strains of four Trichoderma species were able to enter glandular trichomes during the colonization of cacao stems where they survived surface sterilization and could be re-isolated. The penetration of cacao trichomes may provide the entry point for Trichoderma species into the cacao stem allowing systemic colonization of this tissue.

Adding value to cocoa (Theobroma cacao L.) germplasm information with domestication history and admixture mapping.

A sound understanding of crop history can provide the basis for deriving novel genetic information through admixture mapping. We confirmed this, by using characterization data from an international collection of cocoa, collected 25 years ago, and from a contemporary plantation. We focus on the trees derived from three centuries of admixture between Meso-American Criollo and South American Forastero genomes. In both cacao sets of individuals, linkage disequilibrium extended over long genetic distances along chromosome regions, as expected in populations derived from recent admixture. Based on loose genome scans, genomic regions involved in useful traits were identified. Fifteen genomic regions involved in seed and fruit weight variation were highlighted. They correspond to ten previously identified QTLs and five novel ones. Admixture mapping can help to add value to genetic resources and thus, help to encourage investment in their conservation.

Mapping quantitative trait loci for bean traits and ovule number in Theobroma cacao L.

Quantitative trait loci (QTL) mapping for bean traits and the number of ovules per ovary was carried out in cocoa (Theobroma cacao L.) using three test-cross progenies derived from crosses between a lower Amazon Forastero male parent (Catongo) and three female parents: one upper Amazon Forastero (IMC78) and two Trinitario (DR1 and S52). RFLP (restriction fragment length polymorphism), microsatellite, and AFLP (amplified fragment lengthpolymorphism) markers were used for mapping. Between one and six QTL for bean traits (length, weight, and shape index) and one and four QTL for the number of ovules per ovary were detected using composite interval mapping (CIM). Individual QTL explained between 5 and 24% of the phenotypic variation. QTL clusters were identified on several chromosomes, but particularly on chromosome 4. QTL related to bean traits were detected in the same region in both Trinitario parents and in a close region in the upper Amazon Forastero parent. In reference to a previous diversity study where alleles specific to Criollo and Forastero genotypes were identified, it was possible to speculate on the putative origin (Criollo or Forastero) of favorable QTL alleles segregating in both Trinitario studied.

Composition of cocoa shell fat as related to cocoa butter.

The physical and chemical constants of cocoa shell fat (a by-product resulted during the production of cocoa butter at chocolate factories) were almost identical with those of cocoa butter obtained from the same cocoa beans except for their high acid value. Shell fat contained more amount of phospholipid content (as cephalin) than cocoa butter. The lipid classes were almost the same in cocoa butter and shell fat, however, the latter contained an unidentified constituent which was not found in cocoa butter. The fatty acids were determined quantitatively by GLC, and the results showed that the predominant acids in cocoa butter were palmitic, and oleic. Less amounts of capric, myristic, palmitoleic and linoleic were found in cocoa butter, whereas more amounts of these acids were found in shell fat. Cocoa butter gave higher values of stearic and myristic acids than those of shell fat. Seventeen compounds were detected by GLC in the unsaponifiable matter of both cocoa butter and shell fat from which eight were identified as C30 hydrocarbon, C32 hydrocarbon, squalene, alpha-tocopherol, cholesterol, campsterol, stigmasterol and beta-sitosterol in the two samples. The sterols were determined quantitatively, and it was found that the predominant sterol in cocoa butter and shell fat was B-sitosterol. Cocoa butter contained higher values of stigmasterol than that of shell fat, which contained increasing values of campsterol, low values of cholesterol were found in both samples. Stability of cocoa butter and shell fat towards oxidative rancidity at 100 degrees C was the same (10.5 hrs).