Soil amendments to reduce cadmium in cacao (Theobroma cacao L.): A comprehensive field study in Ecuador.

The new EU regulations on maximum levels of cadmium (Cd) in cacao products sparked research on countermeasures to reduce Cd concentrations in cacao beans. This study was set up to test the effects of soil amendments in two established cacao orchards (soil pH 6.6 and 5.1) in Ecuador. Soil amendments included: 1) agricultural limestone at 2.0 and 4.0 Mg ha-1 y-1, 2) gypsum at 2.0 and 4.0 Mg ha-1 y-1 and 3) compost at 12.5 and 25 Mg ha-1 y-1, all amendments were applied at the surface during two subsequent years. Lime application increased the soil pH by one unit down to 20 cm depth. On the acid soil, leaf Cd concentrations decreased by lime application and the reduction factor gradually rose to 1.5 after 30 months. No effects of liming or gypsum on leaf Cd was found in the pH neutral soil. Compost application in the pH neutral soil reduced leaf Cd concentration with factor 1.2 at 22 months but that effect was absent at 30 months after application. Bean Cd concentrations were unaffected by any of the treatments at 22 months after application (acid soil) or 30 months (pH neutral soil) suggesting that any treatment effects on bean Cd might be even more delayed than in leaves. Soil columns experiments in the laboratory showed that mixing lime with compost largely enhanced the depth of lime penetration compared to lime only. Compost + lime reduced 10-3 M CaCl2 extractable Cd in soil without lowering extractable Zn. Our results suggest that soil liming has the potential to lower Cd uptake in cacao in the long term in acid soils and that the compost + lime treatment should be tested at field scale to accelerate the effects of the mitigation.

An investigation of zinc isotope fractionation in cacao (Theobroma cacao L.) and comparison of zinc and cadmium isotope compositions in hydroponic plant systems under high cadmium stress.

This study aims to establish whether zinc (Zn) and cadmium (Cd) share similar physiological mechanisms for uptake and translocation in cacao plants (Theobroma cacao L.). Multiple-collector ICP-MS was used to determine the Zn stable isotope compositions in the roots, stems and leaves of 19 diverse cacao genotypes grown in hydroponics with 20 µmol L-1 CdCl2. Additional plants of one genotype were grown in hydroponic solutions containing lower Cd concentrations (0 and 5 µmol L-1 added CdCl2). Regardless of the Cd concentration used in the exposures, the Zn stable isotope compositions show the same systematic patterns in plant organs, with δ66Znroot > δ66Znstem > δ66Znleaf (δ66Zn denotes relative differences in 66Zn/64Zn ratios in parts per thousand). The mean Zn stable isotope fractionation between the plants and the hydroponic solutions was ε66Znuptake = -1.15 ± 0.36‰ (2SD), indicating preferential uptake of isotopically light Zn by plants from the hydroponic solution. The mean  stable isotope fractionation factor associated with translocation of Zn from roots to shoots, ε66Znseq-mob = + 0.52 ± 0.36‰ (2SD), shows that isotopically heavy Zn is preferentially sequestered in the cacao roots, whilst isotopically light Zn is mobilised to the leaves. A comparison with the Cd stable isotope compositions of the same plants shows that both isotopically light Zn and Cd are preferentially taken up by cacao plants. In contrast to Zn, however, the cacao roots retain isotopically light Cd and transfer isotopically heavy Cd to the leaves.

Omics approaches to understand cocoa processing and chocolate flavor development: A review.

The global market of chocolate has increased worldwide during the last decade and is expected to reach a value of USD 200 billion by 2028. Chocolate is obtained from different varieties of Theobroma cacao L, a plant domesticated more than 4000 years ago in the Amazon rainforest. However, chocolate production is a complex process requiring extensive post-harvesting, mainly involving cocoa bean fermentation, drying, and roasting. These steps have a critical impact on chocolate quality. Standardizing and better understanding cocoa processing is, therefore, a current challenge to boost the global production of high-quality cocoa worldwide. This knowledge can also help cocoa producers improve cocoa processing management and obtain a better chocolate. Several recent studies have been conducted to dissect cocoa processing via omics analysis. A vast amount of data has been produced regarding omics studies of cocoa processing performed worldwide. This review systematically analyzes the current data on cocoa omics using data mining techniques and discusses opportunities and gaps for cocoa processing standardization from this data. First, we observed a recurrent report in metagenomics studies of species of the fungi genus Candida and Pichia as well as bacteria from the genus Lactobacillus, Acetobacter, and Bacillus. Second, our analyzes of the available metabolomics data showed clear differences in the identified metabolites in cocoa and chocolate from different geographical origin, cocoa type, and processing stage. Finally, our analysis of peptidomics data revealed characteristic patterns in the gathered data including higher diversity and lower size distribution of peptides in fine-flavor cocoa. In addition, we discuss the current challenges in cocoa omics research. More research is still required to fill gaps in central matter in chocolate production as starter cultures for cocoa fermentation, flavor evolution of cocoa, and the role of peptides in the development of specific flavor notes. We also offer the most comprehensive collection of multi-omics data in cocoa processing gathered from different research articles.

Characterization of selected dietary fibers microparticles and application of the optimized formulation as a fat replacer in hazelnut spreads.

The present work demonstrates the application of the spray drying technique to produce microparticulates of different dietary fibers with particle sizes<10 µm. It examines their role as potential fat replacers for hazelnut spread creams. Optimization of a dietary fiber formulation containing inulin, glucomannan, psyllium husk, and chia mucilage to obtain high viscosity, water holding capacity, and oil holding capacity was conducted. Microparticles containing 46.1, 46.2, and 7.6 weight percentages of chia seed mucilage, konjac glucomannan, and psyllium husk showed a spraying yield of 83.45 %, a solubility of 84.63 %, and viscosity of 40.49 Pas. When applied to hazelnut spread creams, microparticles substituted palm oil by 100 %; they produced a product with a total unsaturated and saturated fat reduction of 41 and 77 %, respectively. An increase in dietary fibers of 4 % and a decrease in total calories of 80 % were also induced when compared with the original formulation. Hazelnut spread with dietary fiber microparticles were preferred by 73.13 % of the panelist in the sensory study due to an enhancement in brightness. The demonstrated technique could be used to increase the fiber content while decreasing the fat content in some commercial products, such as peanut butter or chocolate cream.

Chloroplast/thylakoid-rich material: A possible alternative to the chemically synthesised flow enhancer polyglycerol polyricinoleate in oil-based systems.

Chloroplasts are abundant organelles in a diverse range of plant materials; they are predominantly composed of multicomponent thylakoid membranes which are lipid and protein rich. Intact or unravelled thylakoid membranes should, in principle, have interfacial activity, but little has been published on their activity in oil-in-water systems, and nothing on their performance on an oil continuous system. In this work different physical methods were used to produce a range of chloroplast/thylakoid suspensions with varying degrees of membrane integrity. Transmission electron microscopy revealed that pressure homogenisation led to the greatest extent of membrane and organelle disruption compared to less energy intensive preparation methods The ability of the derived materials to modulate the flow behaviour of a chocolate model system (65% (w/w) sugar/ sunflower oil (natural amphiphiles removed) suspension) was investigated by acquiring rheological parameters. All chloroplast/thylakoid preparations reduced yield stress, apparent viscosity, tangent flow point and cross over point in a concentration-dependent fashion, although not as significantly as polyglycerol polyricinoleate applied at a commercially relevant concentration in the same chocolate model system. Confocal laser scanning microscopy confirmed presence of the alternative flow enhancer material at the sugar surfaces. This research reveals that low-energy processing methods that do not extensively disrupt thylakoid membranes are applicable to generating materials with marked capacity to affect the flow behaviour of a chocolate model system. In conclusion, chloroplast/thylakoid materials hold strong potential as natural alternatives to synthetic rheology modifiers for lipid-based systems such as PGPR.

Screening for pesticide residues in cocoa (Theobroma cacao L.) by portable infrared spectroscopy.

Rapid assessment of pesticide residues ensures cocoa bean quality and marketability. In this study, a portable FTIR instrument equipped with a triple reflection attenuated total reflectance (ATR) accessory was used to screen cocoa beans for pesticide residues. Cocoa beans (n = 75) were obtained from major cocoa growing regions of Peru and were quantified for pesticides by gas chromatography (GC) or liquid chromatography (LC) coupled with mass spectrometry (MS). The FTIR spectra were used to detect the presence of pesticides in cocoa beans or lipid fraction (butter) by using a pattern recognition (Soft Independent Modeling by Class Analogy, SIMCA) algorithm, which produced a significant discrimination for cocoa nibs (free or with pesticides). The variables related to the class grouping were assigned to the aliphatic (3200-2800 cm-1) region with an interclass distance (ICD) of 3.3. Subsequently, the concentration of pesticides in cocoa beans was predicted using a partial least squares regression analysis (PLSR), using an internal validation of the PLRS model, the cross-validation correlation coefficient (Rval = 0.954) and the cross-validation standard error (SECV = 14.9 mg/kg) were obtained. Additionally, an external validation was performed, obtaining the prediction correlation coefficient (Rpre = 0.940) and the standard error of prediction (SEP = 16.0 µg/kg) with high statistical performances, which demonstrates the excellent predictability of the PLSR model in a similar real application. The developed FTIR method presented limits of detection and quantification (LOD = 9.8 µg/kg; LOQ = 23.1 µg/kg) with four optimum factors (PC). Mid-infrared spectroscopy (MIR) offered a viable alternative for field screening of cocoa.

Effectiveness of various cacao pod husk extraction byproducts in promoting growth and immunocompetence in Litopenaeus vannamei.

Extracts from plant products can promote growth, can act as immunostimulants, and have antibacterial and antiparasitic properties. These extracts can be used as alternatives to the chemical treatments commonly used to prevent and control disease in aquatic species. Research on the subject has focused on identifying invasive plants or agricultural waste products that can be used as immunostimulants. The present study further identified an optimal means of extracting pectin from cacao pod husks to promote growth performance and immunocompetence in Litopenaeus vannamei that would both reduce production costs and enable waste recycling. The byproducts of pectin extraction from cacao pod husks, that is, dried cacao pod husk powder (DCP), steamed DCP (sDCP), hot water-treated cacao pod husk powder (HCP), hot water-treated cacao pod husk supernatant (HCS), and cacao pod husk pectin (CPH pectin), were used to create five experimental diets, which were administered to five groups. The control group was fed a basal diet. The growth and immunocompetence of the shrimp were determined after 30, 60, 90 and 120 days of feeding. To identify the most cost-effective means of obtaining dried cacao pod husks, this study firstly determined the costs and effectiveness of the sun-drying, dehumidification, and heated-wind drying techniques. According to the results of growth performance, the CPH pectin group had higher survival but lower weight gain than the DCP, sDCP, HCP, and HCS groups did. At 30, 60, and 90 days, the clearance efficiency of the experimental groups was higher than that of the control group. At 60 days, the experimental groups had significantly higher phagocytic activity than the control group did. However, at 30 and 90 days the HCP, HCS and CPH pectin group had higher phagocytic activity. The total hemocyte count, differential hemocyte count, phenoloxidase activity, and respiratory bursts of the CPH pectin group were higher at 30 days but the same as those of the control group at 60 and 90 days. After 120 days of feeding trial, the resistance of L. vannamei fed with diets containing byproducts of pectin extraction from cacao pod husks significantly enhanced compared to that in BD group when they were infected with Vibrio aliginolyticus for 168 h, and the related higher survival rate can be observed in HCP, HCS and CPH pectin groups. The study findings suggest that diet-administered HCP and HCS have long-term immunostimulant potential and that CPH pectin has potential in the early stages of feeding. In addition, when heated air drying was employed, a moisture level of below 10% was obtained within 12 h. The results of this study indicate that adding HCP obtained from heated air-dried cacao pod husks to the feed of L. vannamei is the most cost-effective and sustainable means of promoting long-term growth performance and immunocompetence in the species.

Short-Term Intake of Theobroma grandiflorum Juice Fermented with Lacticaseibacillus rhamnosus ATCC 9595 Amended the Outcome of Endotoxemia Induced by Lipopolysaccharide.

Endotoxemia is a condition caused by increasing levels of lipopolysaccharide (LPS) characterized by an impaired systemic response that causes multiple organ dysfunction. Lacticaseibacillus rhamnosus ATCC 9595 is a strain with probiotic potential which shows immunomodulatory properties. The incorporation of this bacterium in food rich in bioactive compounds, such as cupuaçu juice (Theobroma grandiflorum), could result in a product with interesting health properties. This work evaluated the effects of the oral administration of cupuaçu juice fermented with L. rhamnosus on the outcome of LPS-induced endotoxemia in mice. C57BL/6 mice (12/group) received oral doses (100 µL) of saline solution and unfermented or fermented cupuaçu juice (108 CFU/mL). After 5 days, the endotoxemia was induced by an intraperitoneal injection of LPS (10 mg/kg). The endotoxemia severity was evaluated daily using a score based on grooming behavior, mobility, presence of piloerection, and weeping eyes. After 6 h and 120 h, the mice (6/group) were euthanized for analysis of cell counts (in peritoneal lavage and serum) and organ weight. L. rhamnosus grew in cupuaçu juice and produced organic acids without the need for supplementation. The bacteria counts were stable in the juice during storage at 4 °C for 28 days. The fermentation with L. rhamnosus ATCC 9595 changed the metabolites profile of cupuaçu juice due to the biotransformation and enhancement of some compounds. In general, the administration of L. rhamnosus-fermented juice allowed a significant improvement in several characteristics of endotoxemic status (weight loss, hypothermia, severity index, cell migration). In addition, treatment with fermented juice significantly reduced the weight of the spleen, liver, intestine, and kidneys compared to the saline-treated endotoxemic group. Taken together, our data show that short-term intake therapy of cupuaçu juice fermented with L. rhamnosus ATCC 9595 can reduce systemic inflammation in an experimental model of LPS-induced endotoxemia in mice.

Rapid determination of the shell content in cocoa products using FT-NIR spectroscopy and chemometrics.

The determination of the cocoa shell content is of interest because a high shell content causes a reduction in the quality of cocoa products. Consequently, the aim of the present study was the development of a routinely applicable method for the quantitation of shell material in cocoa nibs. For this, 51 fermented cocoa samples of different varieties from 14 cocoa growing countries covering the crop years 2012-2017 were acquired. Admixtures of cocoa nibs with shell material were prepared in a range of 0-20% cocoa shell and subsequently analysed by Fourier transform near-infrared spectroscopy (FT-NIRS). Support vector machine regression models were created, which enabled the prediction of the cocoa shell content in a mixing ratio range of 0-20% with an RMSE of 2.05% and a R2 of 0.88 and in a range of 0-10% with an RMSE of 1.70% and a R2 of 0.72. This predictive capability suggests that the presented method is suitable for rapid determination of cocoa shell content in cocoa nibs. In addition, it was demonstrated that the method is applicable to other relevant cocoa matrices, as the prediction of the shell content of several industrial cocoa masses by the FT-NIRS-based model showed good consistency with the prediction by liquid chromatography-mass spectrometry. This emphasizes that FT-NIRS combined with chemometrics has great potential for the determination of cocoa shell content in cocoa nibs and cocoa masses in routine analysis, such as incoming inspection.

Cocoa Flavanol Supplementation and the Effect on Insulin Resistance in Females Who Are Overweight or Obese: A Randomized, Placebo-Controlled Trial.

There is interest in the impact that dietary interventions can have on preventing the transition from insulin resistance to type 2 diabetes, including a suggestion that the bioactive components of cocoa may enhance fasting insulin sensitivity. However, a role for cocoa flavanols (CF) in reducing insulin resistance in the insulin-stimulated state, an important risk factor for cardiovascular disease, is unresolved. This study investigated whether CF consumption improved whole-body insulin-mediated glucose uptake ('M') in females with overweight/obesity, using a randomized, double-blinded, placebo-controlled, parallel-group design. Thirty-two premenopausal females (19-49 years; 27-35 kg·m-2) with elevated HOMA-IR (HOMA-IR >1.5) supplemented their habitual diet with two servings/day of a high-flavanol cocoa drink (HFC; 609 mg CF/serving; n = 16) or low-flavanol cocoa drink (LFC; 13 mg CF/serving; n = 16) for 4 weeks. Assessment of HOMA-IR and 'M' during a 3-h, 60 mIU insulin·m-2·min-1 euglycemic clamp was performed before and after the intervention. Data are the mean (SD). Changes to HOMA-IR (HFC -0.003 (0.57); LFC -0.0402 (0.86)) and 'M' (HFC 0.99 (7.62); LFC -1.32 (4.88) µmol·kg-1·min-1) after the intervention were not different between groups. Four weeks' consumption of ~1.2 g CF/day did not improve indices of fasting insulin sensitivity or insulin-mediated glucose uptake. A recommendation for dietary supplementation with cocoa flavanols to improve glycemic control is therefore not established.

Discriminating Aroma Compounds in Five Cocoa Bean Genotypes from Two Brazilian States: White Kerosene-like Catongo, Red Whisky-like FL89 (Bahia), Forasteros IMC67, PA121 and P7 (Pará).

Fine-grade cocoa beans are characterized by their great organoleptic quality. Brazilian cocoa producers increasingly privilege organoleptic quality over yield. Catongo and FL89, fine Brazilian cocoa genotypes from Bahia, are characterized by particular flavors (respectively, kerosene and whisky). The beans IMC67, PA121, and P7 from the state of Pará are genotypes that have high resistance to diseases. Solvent-assisted flavor evaporation (SAFE) aroma extraction was used here to identify and quantify the volatile compounds discriminating these genotypes. The results show that the kerosene aroma of Catongo is likely due to the presence, at high levels, of ethyl acetate and isobutyl acetate. On the other hand, ethyl benzoate, heptanoate, and octanoate, trans-2-nonenal, 1-octen-3-ol, and 3-methylbutanol could play a key role in the whisky notes of FL89. Heptan-2-ol, heptan-2-one, nonan-2-one, linalool (although still more concentrated in IMC67 from Pará state), benzaldehyde, and phenylacetaldehyde also discriminate these beans. Other compounds, although not discriminating, appear important in determining their aromatic quality. The PCA showed that cocoa from Pará state formed a cluster due to similar aromas, while FL89 was the most distinct among the genotypes. Beans from Brazil show great potential and diversity for the fine cocoa market.

Estimated Dietary Fluoride Intake by 24-Month-Olds from Chocolate Bars, Cookies, Infant Cereals, and Chocolate Drinks in Brazil.

The use of fluoride (F) in the prevention of dental caries is established. However, a high amount of F intake during tooth development can cause dental fluorosis The aim of this study was to analyze variations in F concentrations in chocolate bars (CB), chocolate cookies (CC), infant cereals (IC), and chocolate milk drinks (CD) to determine the daily intake of F from different sources by children at the age of risk for developing dental fluorosis. Distinct brands of CB, CC, IC, and CD were analyzed. Fluoride was separated by hexamethyldisiloxane-facilitated diffusion. Analysis was made in triplicate with an F ion-specific electrode. F ingestion (mg/kg body weight) was evaluated with the suggested consumption (0.05-0.07 mg/kg/day) for children aged 24 months (12 kg). The concentrations for all the analyzed products ranged from 0.025 to 1.827 µg/g F. The mean (range) F concentrations were CB= 0.210 ± 0.205 µg/g (0.073-0.698, n = 8), CC = 0.366 ± 0.416 µg/g (0.320-1.827, n = 9), IC = 0.422 ± 0.395 µg/g (0.073-1.061, n = 5), and CD = 0.169 ± 0.170 µg/mL (0.025-0.443, n = 12). The products that had the highest concentration in the categories CB, CC, IC, and CD, respectively, were Nescau-Ball (0.698 µg/g), Passatempo (1.827 µg/g), Milnutri (1.061 µg/g), and Toddynho (0.443 µg/mL). The consumption of only one unit of Toddynho (CD) is equivalent to more than 11% of the maximum suggested daily intake for a 24-month-old child (0.07 mg/kg body weight). When one product from each category is consumed together only once a day, this consumption is equivalent to approximately 24% of the suggested daily intake of fluoride for a 24-month-old child. The presence of high levels of fluoride in certain products suggests that they play a significant role in overall fluoride intake. It is crucial to closely monitor the fluoride content of food and drinks that are consumed by children who are at risk for dental fluorosis, and for product labels to clearly display the fluoride concentrations.

Rationale and design of an ancillary study evaluating the effects of the cocoa flavanol and/or multivitamin interventions on falls and physical performance outcomes in the COcoa Supplement and Multivitamin Outcomes Study (COSMOS).

BACKGROUND: Falls and decreased physical function increase markedly with age and result in injury, hospitalization, and premature death. Emerging studies show potential benefits of supplemental cocoa extract on physical performance, including grip strength and walking speed in older adults. However, there are no large, long-term randomized controlled trials of effects of supplemental cocoa extract on falls, muscle performance, and/or fall-related injuries. METHODS: The COcoa Supplement and Multivitamin Outcomes Study (COSMOS) is a double-blind, placebo-controlled, 2 × 2 factorial trial investigating effects of supplementation with cocoa extract (500 mg/d, including 80 mg (-)-epicatechin) and/or a multivitamin on prevention of cardiovascular disease and cancer in 21,442 women (≥65 years) and men (≥60 years). COSMOS: Effects on Falls and Physical Performance is an ancillary study to COSMOS that will clarify effects of cocoa extract and/or multivitamin supplementation on falls, physical performance, and incident fracture outcomes in older adults. Injurious fall(s) resulting in healthcare utilization and recurrent falls were regularly assessed by follow-up questionnaires in the overall cohort. Incident fractures were also assessed by annual questionnaires. Circumstances surrounding falls and any fall-related injuries will be confirmed by medical record review. Effects of the interventions on 2-year changes in physical performance measures (grip strength, walking speed, and the Short Physical Performance Battery) will be tested in a clinic sub-cohort (n = 603). CONCLUSION: Results from this ancillary study will determine whether supplemental cocoa extract slows age-related declines in physical performance and decrease injurious and recurrent falls and fall-related injuries and fractures that are major public health problems in older adults.

Physiological and growth responses of cacao to glyphosate exposure.

Herbicide drift phytotoxicity is a problem in plantation crops due to application failures and unfavorable spray conditions. With the increased use of glyphosate in cacao plantations in Ghana, there are concerns about the effect on cacao growth and productivity from doses that potentially could be expected from drift. The aim of this study was to evaluate the physiological and growth response of young cacao plants exposed to glyphosate. Two field experiments were conducted in randomized blocks, with four replications. Glyphosate was applied at rates 0 to 720 g a.e. ha-1. Crop injury, shikimate accumulation, chlorophyll content, quantum efficiency of PSІІ (Fv/Fm), height, and stem diameter were evaluated. Increased glyphosate rates increased crop injury and shikimate accumulation and decreased chlorophyll content, quantum efficiency of PSІІ (Fv/Fm), and plant growth. Glyphosate rates 360 g a.e. ha-1 or higher resulted in >60% foliar injury and more than 10-fold increase in shikimate accumulation. Glyphosate reduced chlorophyll content to <10 and Fv/Fm to <0.35 at the highest rates. Glyphosate rates ≥180 g a.e. ha-1 reduced height and stem diameter of plants and caused reductions in stand count. Thus, cacao showed sensitivity to glyphosate, and severe injury impaired plant growth.

Alkaline Modification of Arabica-Coffee and Theobroma-Cocoa Agroindustrial Waste for Effective Removal of Pb(II) from Aqueous Solutions.

Arabica-coffee and Theobroma-cocoa agroindustrial wastes were treated with NaOH and characterized to efficiently remove Pb(II) from the aqueous media. The maximum Pb(II) adsorption capacities, qmax, of Arabica-coffee (WCAM) and Theobroma-cocoa (WCTM) biosorbents (qmax = 303.0 and 223.1 mg·g−1, respectively) were almost twice that of the corresponding untreated wastes and were higher than those of other similar agro-industrial biosorbents reported in the literature. Structural, chemical, and morphological characterization were performed by FT-IR, SEM/EDX, and point of zero charge (pHPZC) measurements. Both the WCAM and WCTM biosorbents showed typical uneven and rough cracked surfaces including the OH, C=O, COH, and C-O-C functional adsorbing groups. The optimal Pb(II) adsorption, reaching a high removal efficiency %R (>90%), occurred at a pH between 4 and 5 with a biosorbent dose of 2 g·L−1. The experimental data for Pb(II) adsorption on WACM and WCTM were well fitted with the Langmuir-isotherm and pseudo-second order kinetic models. These indicated that Pb(II) adsorption is a chemisorption process with the presence of a monolayer mechanism. In addition, the deduced thermodynamic parameters showed the endothermic (ΔH0 > 0), feasible, and spontaneous (ΔG0 < 0) nature of the adsorption processes studied.

Flavan-3-ol-methylxanthine interactions: Modulation of flavan-3-ol bioavailability in volunteers with a functional colon and an ileostomy.

Flavan-3-ols, including the flavan-3-ol monomer (-)-epicatechin, are dietary bioactives known to mediate beneficial cardiovascular effects in humans. Recent studies showed that flavan-3-ols could interact with methylxanthines, evidenced by an increase in flavan-3-ol bioavailability with a concomitant increase in flavan-3-ol intake-mediated vascular effects. This study aimed at elucidating flavan-3-ol-methylxanthine interactions in humans in vivo by evaluating the specific contributions of theobromine and caffeine on flavan-3-ol bioavailability. In ileostomists, the effect of methylxanthines on the efflux of flavan-3-ol metabolites in the small intestine was assessed, a parameter important to an understanding of the pharmacokinetics of flavan-3-ols in humans. In a randomized, controlled, triple cross-over study in volunteers with a functional colon (n = 10), co-ingestion of flavan-3-ols and cocoa methylxanthines, mainly represented by theobromine, increased peak circulatory levels (Cmax) of flavan-3-ols metabolites (+21 ± 8%; p < 0.05). Conversely, caffeine did not mediate a statistically significant effect on flavan-3-ol bioavailability (Cmax = +10 ± 8%, p = n.s.). In a subsequent randomized, controlled, double cross-over study in ileostomists (n = 10), cocoa methylxanthines did not affect circulatory levels of flavan-3-ol metabolites, suggesting potential differences in flavan-3-ol bioavailability compared to volunteers with a functional colon. The main metabolite in ileal fluid was (-)-epicatechin-3'-sulfate, however, no differences in flavan-3-ol metabolites in ileal fluid were observed after flavan-3-ol intake with and without cocoa methylxanthines. Taken together, these results demonstrate a differential effect of caffeine and theobromine in modulating flavan-3-ol bioavailability when these bioactives are co-ingested. These findings should be considered when comparing the effects mediated by the intake of flavan-3-ol-containing foods and beverages and the amount and type of methylxanthines present in the ingested matrixes. Ultimately, these insights will be of value to further optimize current dietary recommendations for flavan-3-ol intake. CLINICAL TRIAL REGISTRATION NUMBER: This work was registered at clinicaltrials.gov as NCT03526107 (study part 1, volunteers with functional colon) and NCT03765606 (study part 2, volunteers with an ileostomy).

Differential Orbitofrontal Cortex Responses to Chocolate Images While Performing an Approach-Avoidance Task in the MRI Environment.

Background: Chocolate is one of the most frequently craved foods, and it often challenges self-regulation. These cravings may be underpinned by a neural facilitation of approach behavior toward chocolate. This preregistered study investigated the behavioral and neural correlates of such a bias using functional magnetic resonance imaging (fMRI) and reaction times (RTs). Methods: A total of n = 30 frequent chocolate eaters performed a relevant-feature approach-avoidance task (AAT) in the MRI scanner using buttons to enlarge (approach) or to shrink (avoid) pictures of chocolate and inedible control objects. We tested (a) whether implicit RT-based approach biases could be measured in a supine position in the scanner, (b) whether those biases were associated with activity in reward-related brain regions such as the insula, amygdala, striatum, and orbitofrontal cortex (OFC), and (c) whether individual RT-based bias-scores correlated with measures of chocolate craving. Results: Behaviorally, we found a highly reliable approach bias toward chocolate, defined by faster RTs in the compatible conditions (approach chocolate, avoid objects) compared to the incompatible conditions (avoid chocolate, approach objects). Neurally, this compatibility effect involved activity in the left medial OFC, a neural response that was positively correlated with individual approach bias scores. Conclusions: This study shows that the relevant feature AAT can be implemented in an fMRI setting in a supine position using buttons. An approach bias toward chocolate seems related to medial OFC activation that might serve to devalue chocolate when it has to be avoided. Our demonstration of neural and behavioral approach biases for chocolate underscores the need for stimulus-specific cognitive trainings to support healthy consumption and successful self-regulation.

Accumulation and decomposition of cultural residues of Theobroma grandiflorum, Paullinia cupana, Bixa orellana and forest in the southern region of Amazonas.

The litter deposited on the soil surface at various stages of decomposition is important for primary productivity that impacts the microbial communities and soil carbon storage. The objective of this study was to evaluate the accumulation and decomposition of cultural residues of Theobroma grandiflorum (Willd. ex. Spreng) Schum, Paullinia cupana (Mart.) Ducke, Bixa orellana L., and forest in the Amazon region. The study was carried out in the São Francisco settlement, Canutama in the south of Amazonas, in a randomized block experimental design, and the treatments consisted of four areas with different crops: 1 - P. cupana; 2 - T. grandiflorum; 3 - B. orellana; 4 - Native woodland area (forest), in time subdivided plots: 7, 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, and 330 days after the distribution of the bags in the field, all with four repetitions. To evaluate the contribution and fractions of litter, conical collectors were used in each area, and collections were performed monthly in the period from March 2020 to February 2021. The estimate of the decomposition rate of the litter was done by quantifying the loss of mass, using litter bags, which allow for a direct analysis of the rate of decay over time. The forest and P. cupana environments presented the highest litter production, and greater deposition when compared to environments cultivated with T. grandiflorum and B. orellana. The forest and B. orellana areas showed the highest speed of decomposition, while the opposite situation occurred under T. grandiflorum and P. cupana cultivation.

The contribution of cacao consumption to the bioaccessible dietary cadmium exposure in the Belgian population.

Since 2019, EU limits apply to cadmium (Cd) concentrations in cacao-derived food products. The dietary risk assessment leading to that regulation used consumption surveys aggregated to a limited number of chocolate product categories and did not consider differences in Cd bioaccessibility. Here, the cacao-related dietary Cd exposure in the Belgian population was estimated with higher resolution and accounting for bioaccessibility. A food frequency questionnaire and a 24-h recall (N = 2055) were set up for the Belgian population, in combination with ICP-MS analysis of a large subset of cacao-containing products (N = 349). Both the average chocolate consumption (28 g day-1) and the relative contribution of chocolate to the total dietary Cd exposure (7-9%) were higher than previously estimated for the Belgian population, probably because of some selection bias towards chocolate consumers in the cohort. The Cd bioaccessibility in chocolate products was a factor 5 (cacao powder) and 2 (dark chocolate) lower compared to wheat flour, suggesting lower bioavailability in chocolate than in wheat, which is a main contributor to dietary Cd. This study suggests that Cd intake from cacao consumption has been underestimated because of hidden cacao in non-chocolate food categories but, in contrast, may have overestimated the true exposure because of lower bioavailability compared to the main foodstuffs contributing to Cd exposure.

New trends of new psychoactive substances (NPS)-infused chocolate: Identification and quantification of trace level of NPS in complex matrix by GC-MS and NMR.

For the first time, the identification and quantification of trace level of new psychoactive substances (NPS) in a complex chocolate matrix have been reported. Since the beginning of 2022, suspected NPS-infused chocolate samples confiscated in inbound packages have been continuously sent to our laboratory for analysis. The qualitative gas chromatography-mass spectrometry (GC-MS) results were verified by 1H nuclear magnetic resonance (1H NMR) and 19F NMR to distinguish between potential aromatic isomers. A total of 11 NPS including deoxymethoxetamine, 3-OH-PCP, 6-APB, 4-APB, 4-OH-MiPT, 3-FEA, 2-FEA, 3-MMC, bromazolam, 2-FDCK, and ADB-BUTINACA were detected in 65 seized chocolate samples. A general 1H quantitative NMR (1H qNMR) method for quantification of 297 types of NPS in complex chocolate matrixes was devised for the first time after rigorous analysis of various critical features of merit, including suitable deuterated solvent, internal standard, quantitative peaks, and instrument acquisition parameters. Validation of the method using six different types of NPS afforded limits of detection of 0.05-0.1 mg/mL, limits of quantification of 0.01-0.03 mg/mL, repeatability and reproducibility lower than 0.5% and 3.6%, recoveries of 91.7%∼104.4%, and absence of matrix effect. The quantitative analysis of 65 seized chocolate samples by 1H qNMR and 19F qNMR showed that the content of NPS was in the range of 0.5 mg/g∼44.1 mg/g. Generally, the developed qNMR method was simple, fast, precise, and can be performed without reference materials of NPS. Since the type and content of NPS are relatively random, chocolate consumers will face huge health risks. Therefore, this new trend of NPS-infused chocolate deserves and requires more attention from national NPS monitoring departments as well as forensic laboratories.