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In Mexico, the prevalence of Salmonella enterica in low-water-activity foods and its link to outbreaks are unknown. The aim of this study was to determine the microbiological profile and the prevalence of S. enterica in several low-water-activity foods, including peanuts, pecans, raisins, sun-dried tomatoes, and chocolate sprinkles, purchased in retail establishments in Querétaro, Mexico. Seventy samples of each food item sold in bulk were purchased. Aerobic plate count, molds, yeasts, total coliforms, Escherichia coli, and Staphylococcus aureus were quantified in 10-g samples. The prevalence of S. enterica in 25-g samples was determined. From positive samples, S. enterica isolates (60) were characterized based on their antimicrobial susceptibility to 14 antibiotics, the presence-absence of 13 virulence genes, and serotype. The concentration of aerobic plate count, molds, yeasts, total coliforms, and E. coli ranged from 3.1 to 5.2 log CFU g-1, from 2.0 to 2.4 log CFU g-1, from 2.0 to 3.0 log CFU g-1, from 0.6 to 1.1 log most probable number (MPN) g-1, and from 0.5 to 0.9 log MPN g-1, respectively. S. aureus was not detected in any sample (10 CFU g-1). The prevalence of S. enterica in chocolate sprinkles, raisins, peanuts, pecans, and sun-dried tomatoes was 26, 29, 31, 40, and 52%, respectively. Most isolates (68.3%) were resistant to at least one antibiotic. Chromosome-associated virulence genes were found in all isolates, and only one strain had sopE, and 98.3% of the isolates were grouped in the same virulotype. Among the isolates, the most frequent serotype was Tennessee (51 of 60). According to the characteristics evaluated, we grouped the isolates into 24 clusters. The elevated prevalence of S. enterica highlights the role of low-water-activity food items sold in bulk at markets as potential vehicles for pathogen transmission. Regardless of the low variability among S. enterica isolates, their characterization could be helpful to elucidate which strains are circulating in these foods for improving epidemiological surveillance.

Chocolate is one of the most popular sweets in the world. In recent years, the bean-to-bar process for chocolate production has attracted global attention. Bean-to-bar is a method of managing the whole production process from cocoa beans to chocolate bars, including single-origin chocolate (SOC). Many manufacturers aim to produce high-quality chocolate to maximize the flavor of cocoa beans. However, chocolate compounds are very complex due to many processes, and there are a limited number of studies on the SOC produced from the bean-to-bar process. Therefore, understanding the effects of processing is important for the growth of the chocolate industry. The objective of this study was to investigate the processing effect on the component changes of SOC during the bean-to-bar process. In this study, the component changes during the bean-to-bar process were monitored using gas chromatography/mass spectrometry (GC/MS). Then, the characteristics of SOC from five regions in Indonesia were further investigated. Lastly, the component profiles were combined with the data obtained from sensory evaluation. Our results showed that the influence of the manufacturing process was greater than that of the difference in the cocoa production area. Moreover, 1-pentanol, raffinose, and heptanoic acid were correlated with sweetness and dairy flavor, whereas glutamic acid, tartaric acid, 3-methyl-2-butanone, mannitol, and ethyl cinnamate were correlated with bitterness, astringency, and cocoa flavor, which were shown to be affected by fermentation, roasting, and sugar addition. This information might provide a basis for improving the chocolate production process and its quality related to the component profiles.

Chocolate flavors vary depending on the origin of the cocoa beans used. Differences in soil, microorganisms, and environmental factors contribute to the formation of flavor precursors in cocoa beans. During cocoa bean fermentation, environmental temperature has been shown to alter metabolite concentrations. However, the correlation between the metabolite profile of cocoa beans and the temperature of their region of origin has not been fully defined. In this study, the metabolite profiles of Indonesian cocoa beans of various origins were evaluated using gas chromatography/mass spectrometry-based analysis, and were found to differ depending on the origin of the bean. Subsequently, the correlation between metabolite profile and environmental temperature of the origin was assessed using orthogonal projection to latent structure regression (OPLS-R) analysis. The analysis revealed that seven metabolites were associated with temperature: γ-aminobutyric acid (GABA), ethanolamine, glycerol, isocitric acid + citric acid, succinic acid, malic acid, and saccharic acid. The findings of this study will be valuable to chocolate industries for the production of single-origin chocolates.

BACKGROUND Coffee consumption is associated with a reduced risk of several chronic diseases in a dose-dependent manner. Chronic intake results in the transient appearance of bioactive phenolic metabolites in the circulatory system. However, there is a lack of information on the impact of different patterns of coffee consumption on plasma and urinary profiles of phenolic metabolites. OBJECTIVES Plasma and urinary phenolic metabolites were investigated following regular consumption of different daily dosages of coffee or cocoa-based products containing coffee (CBPCC) under a real-life setting. METHODS A repeated-dose, randomized, crossover human intervention was conducted with 21 healthy volunteers. For 1 mo, participants consumed 1) 1 cup of coffee (1C), 2) 3 cups of coffee (3C), or 3) 1 cup of coffee + 2 CBPCC twice daily (PC). Plasma and urine samples were collected over a 24-h period after each treatment. The nutrikinetics and urinary excretion of native, human phase II, and colonic metabolites were assessed. RESULTS A total of 51 (poly)phenolic metabolites were quantified, with 41 metabolites being strictly related to coffee consumption. Significant differences were observed among treatments for most of the metabolites. The metabolites present in the highest amounts were the hydroxycinnamate, phenylpropanoic acid, benzaldehyde, and benzene classes, along with (-)-epicatechin and phenyl-γ-valerolactone derivatives after PC treatment. Daily average concentrations did not exceed 200 nmol/L and were

The endocannabinoid system plays an important role in multiple behavioral responses due to its wide distribution in the central nervous system. The cannabinoid CB1 receptor was associated to the loss of behavioral control over food intake occurring during food addiction. The cannabinoid CB2 receptor (CB2R) is expressed in brain areas canonically associated with addictive-like behavior and was linked to drug-addictive properties. In this study, we evaluated for the first time the specific role of the CB2R in food addiction by using a well-validated operant mouse model of long-term training to obtain highly palatable food. We have compared in this model the behavioral responses of wild-type mice, mutant mice constitutively lacking CB2R, and transgenic mice overexpressing CB2R. The lack of CB2R constitutes a protective factor for the development of food addiction and the impulsive and depressive-like behavior associated. In contrast, the overexpression of CB2R induces a vulnerable phenotype toward food addiction after long-term exposure to highly palatable chocolate pellets. Relevant transcriptomic changes were associated to resilience and vulnerability to food addiction depending on the genotype, which provides a mechanistic explanation for these behavioral changes. Therefore, CB2R may constitute a potential therapeutic target for the loss of eating control and the comorbid emotional effects associated to food addiction. This work was supported by the Spanish “Ministerio de Ciencia e Innovación (MICIN), Agencia Estatal de Investigación (AEI)” (PID2020-120029GB-I00/MICIN/AEI/10.13039/501100011033, RD21/0009/0019, to RM; the “Generalitat de Catalunya, AGAUR” (2017 SGR-669, to RM; the “ICREA-Acadèmia” (2020) to RM; the “European Commission-DG Research” (PainFact, H2020-SC1–2019-2-RTD-848099, QSPain Relief, H2020-SC1–2019-2-RTD-848068) to RM; the Spanish”la Caixa” Foundation under the project code LCF/PR/HR22/52420017 to R.M., the Spanish “Instituto de Salud Carlos III, RETICS-RTA” (RD16/0017/0020) to RM; the Spanish “Ministerio de Sanidad, Servicios Sociales e Igualdad, Plan Nacional Sobre Drogas” (PNSD- 2021I076, to RM; PNSD- 2019I006, to EMG; Ministerio de Ciencia e Innovación (ERA-NET) PCI2021–122073-2A to EMG; and “Instituto de Salud Carlos III”, Spanish “Ministerio de Ciencia e Innovación” (RD21/0009/0008, “Red de Investigación en Atención Primaria de Adicciones”) to JM.

The influence of under-fermented (UF) cocoa (0 to 65 %) on bioactive amines in chocolate and their in vitro bioaccessibility was investigated. The same amines were found in all treatments; however, treatments were divided into two groups regarding total amines [020 % UF (34 mg/kg) and 35 to 65 % UF (17 mg/kg)] and phenolic levels [lower and higher, respectively]. Serotonin, tyramine, putrescine, cadaverine, agmatine and phenylethylamine were higher in chocolate with ≤ 20 % UF cocoa. Histamine and spermidine were not affected. Digestibility studies indicated that low levels of amines were present in the oral phase. Gastric digestion was effective in releasing tyramine, spermidine and phenylethylamine from conjugates. Serotonin and agmatine were not detected after in vitro digestion of chocolate with ≥ 35 % UF cocoa. Histamine was released during in vitro intestinal digestion. By adding different proportions of UF cocoa during chocolate production, the levels and bioaccessibility of amines can be modulated.

Background Previous studies have shown favorable effects of chocolate products on the cardiovascular system and reported an inverse relationship between chocolate consumption and adverse cardiac events; however, relationships between chocolate eating habits and heart rate variability have not yet been studied. The purpose of the present research was to determine the relationship, if any, between chocolate consumption frequency and heart rate variability. Methods A cross-sectional study was carried out in a group of 98 young females (19–21 years old). The study included the evaluation of chocolate eating habits by a questionnaire and heart rate variability parameters by Finometer and Powerlab as primary outcomes along with measuring anthropometry, blood pressure and fasting blood sugar levels as predictors to account for potential confounding. Results Three chocolate eating patterns were distinguished: “No regular chocolate consumption (n:42), 2–4 servings/week (n:20), 5 or more servings/week (n:36)”. An ANOVA test revealed insignificant differences (p > 0.05) in heart rate variability parameters (time domain: Heart rate, RRms, SDNN, RMSSD; frequency domain: LF, HF, LF/HF) and pressures (systolic, diastolic, pulse, mean arterial) among three groups of individuals having different chocolate eating habits. Relative frequency of chocolate intake did not correlate with any of the cardiovascular, time-domain or frequency-domain parameters of heart rate variability in study participants. Conclusion Based on our results, we conclude that relative frequency of chocolate eating may not affect autonomic regulation of the heart in young females. Also, it may have no correlation with any of the cardiovascular, time-domain or frequency-domain parameters of heart rate variability.

The survival and virulence of Listeria monocytogenes was assessed during storage on three low-moisture foods (LMFs): chocolate liquor, corn flakes, and shelled, dry-roasted pistachios (water activity [aw] of 0.18, 0.27, and 0.20, respectively). The LMFs were inoculated with a four-strain cocktail of L. monocytogenes at 8 log CFU/g, dried, held until the aw stabilized, and then stored at 4°C and 25 to 81% relative humidity (RH) and at 23°C and 30 to 35% RH for at least 336 days. At 4°C, L. monocytogenes remained stable on the LMFs for at least 336 days. At 23°C, L. monocytogenes levels declined on the chocolate liquor, corn flakes, and pistachios at initial rates of 0.84, 0.88, and 0.32 log CFU/g/month, respectively. After 8 months at 23°C, L. monocytogenes levels on the chocolate liquor and corn flakes decreased to below the limit of detection (i.e., 0.48 log CFU/g). Relative populations of each strain were assessed before storage (i.e., day 0) and after 6 and 12 months of storage at 23 and 4°C, respectively. Generally, a decline in the relative level of the serotype 1/2a strain was observed during storage, coupled with the relative increase in other strains, depending on the LMF and storage temperature. The total viable populations of L. monocytogenes determined by the PMAxx quantitative PCR method after >12 months of storage at 4°C were significantly (1.8- to 3.7-log) higher than those obtained by plating on tryptic soy agar with yeast extract. Decreases in the culturable population of L. monocytogenes during storage on the LMFs were the result of both cellular inactivation and transition to a viable-but-nonculturable state. The surviving cells, specifically after long-term storage at 4°C on the chocolate liquor and pistachios, remained infectious and capable of intracellular replication in Caco-2 enterocytes. These results are relevant for predictive modeling used in microbial health risk assessments and support the addition of LMFs to food safety questionnaires conducted during listeriosis outbreaks. HIGHLIGHTS

The presence of bacterial spores in cocoa powders is inevitable due to the cocoa bean fermentation process, during which members of the genera Bacillus and Geobacillus are typically present. Spores are a concern in heat-treated foods when they survive heat treatments and the finished product supports germination, growth, and potentially toxin production. In this study, available methods for the enumeration of total mesophilic and thermophilic spores (TMS and TTS, respectively) were evaluated, leading to the recommendation of one global method specifically for cocoa powders. The proposed method was validated during a ring test on seven selected cocoa powders and applied during routine analyses on commercial powders. The method includes dilution of cocoa powder using buffered peptone water, heating at 80°C for 10 min for TMS and TTS counts, and heating at 100°C for 30 min for a heat-resistant (HR) spore count. Tryptic soy agar is used as a recovery medium with a maximal concentration of cocoa powder of 2.5 mg/mL (to prevent growth inhibition) and a nonnutrient agar overlay to prevent swarming of bacteria. Plates are incubated for at least 72 h at 30°C for recovery of mesophilic bacteria and 55°C for thermophilic bacteria. Suitable alternatives to specific method parameters are provided. Median values of total spore concentrations are low (

Modifications to pathogen detection kits to accomplish simplified protocols with reduced time to results may impact method performance, particularly when combining shortened enrichment times and simplified enrichment procedures. We used Salmonella detection in dark chocolate as a model to test the impact of different enrichment times (minimum and maximum validated times) and procedures on detection of low levels of difficult-to-detect Salmonella strains, for three PCR kits that were AOAC International Performance Tested Method certified for detection of Salmonella spp. in dark chocolate. Initial inclusivity studies with pure cultures showed that all three kits detected 70 of 70 Salmonella spp. strains at 1 log above the theoretical limit of detection, with some strains yielding later cycle threshold values or having variable detection among technical replicates, indicating reduced assay performance for these strains. Based on these data, we selected a S. enterica subsp. enterica serovar Poona strain as well as three non-subsp. enterica strains to test the ability of the three kits to detect Salmonella in dark chocolate inoculated at low levels (0.06 to 1.18 most probable number per 25 g). With primary enrichment in skim milk at 35°C, detection frequency for all assays did not significantly differ from the reference method for both the minimum and maximum validated enrichment times. However, a pilot study that used primary enrichment in buffered peptone water at 42°C yielded significantly fewer positive samples (13 of 80) than were obtained with the U.S. Food and Drug Administration Bacteriological Analytical Manual method using enrichment in skim milk at 35°C (40 of 80 positive samples); strains representing subsp. houtenae and salamae were detected in significantly fewer chocolate samples than enrichment with skim milk. Our data indicate that continued efforts to simplify rapid pathogen detection kits may reduce kit performance in a way that can only be detected with stringent evaluation protocols that are designed to identify kit failure modes. HIGHLIGHTS

Evaluations and intake of unhealthy, palatable food can be targeted via motor response training procedures such as Go/No-Go training (GNG) and Approach-Avoidance training. While evidence especially supports the effectiveness of GNG in changing food intake, both tasks seem to affect evaluations of trained stimuli. Associative accounts explain this devaluation through the formation of associative links with rudimentary appetitive/aversive motivational systems. Alternative models that focus on the resolution of conflict between appetitive stimuli and inhibition, or on inferences about stimulus value through valenced actions, however, may better explain the boundary conditions of motor response training effects. Future research should further test hypotheses derived from these accounts using comparable research protocols to elucidate commonalities and differences between these motor response training tasks.

Gastric distension and detection of macronutrients (calories) in the gut are determinants of satiation and satiety. We tested effects of these variables on body sensations after eating, and their connection with visual-analogue scale (VAS) hunger and fullness ratings. Participants completed VAS ratings and quality and location of body sensations tasks after consumption of milk chocolate (38 g, 200 kcal) versus fresh apple fruit matched for weight (38 g, 20 kcal) and matched for calories (380 g, 200 kcal). Effects of food weight (380 vs 38 g) were large and located predominantly in the abdominal region. They also occupied a greater body area and occurred sooner after eating than effects related to calories (200 vs 20 kcal). The same pattern was apparent in the results from the quality of sensations task. VAS ratings indicated that hunger was affected by food volume and calories, whereas fullness was affected primarily by food volume. Together, these results provide evidence of dissociation of the perceived after-effects of food ingestion related to food volume and food calorie content in humans. Additionally, the studies demonstrate the utility of two rarely used, semi-quantitative tasks, which generate information on the identity, intensity, valence, and location of eating-related sensations.

Origin-based identification and awards are used by craft chocolate makers as cues to signify superior quality and more inclusive practices. However, there is a lack of consumer choice and willingness to pay (WTP) literature that explores the impact of sensory and external cues, particularly award and origin labels, of craft chocolates. Thus, this study aims to combine hedonic and WTP measurements through an experimental auction of three Philippine tablea (cocoa liquor) variants from three origins (i.e., Bohol, Bicol, and Davao) in the Philippines. Information was revealed in a four-round second-price auction: blind condition, award or origin label (ran-domized order), and informed condition. Results indicated that participants were willing to pay a premium to switch their endowed tablea to the three auctioned tablea. Hedonic and WTP measurements resulted in an overall consistency of the hierarchy of preference between tablea variants. Award and origin significantly improved sensory ratings and WTP premiums for the three variants. Econometric analyses revealed that taste positively influenced the propensity to exchange and WTP premiums for all variants. In conclusion, this study found that taste was the most vital factor in choosing tablea, while award and origin labels may be used to further enhance perceived quality and value.

Industrially tempered dark chocolate processed under different conditions was analyzed using Attenuated Total Reflection-Fourier Transform-Infrared spectroscopy (ATR-FT-IR). Spectra were collected during the cooling process right after tempering using two distinct machines and machine operational modes and over short and long storage (1 day and two months, respectively). To interpret the spectra collected, Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) analysis was used. MCR-ALS incorporated a dedicated implementation of hard modeling constraints for the elucidation of the spectral and concentration profiles of chocolate contributions during the monitored process with the aim of reducing ambiguity in the data interpretation. Thus, the spectral signature of the amorphous form of chocolate was constrained to follow a Gaussian shape, in line with previously reported works and with measurements observed at the beginning of the cooling process. Additionally, hard modeling constraints were applied to the concentration profiles linked to the cooling process by implementing the Avrami kinetic model, often postulated to describe thermally induced fat crystal transformations. The application of the hard modeling constraints eased the generation of interpretable solutions and resulted in concentration profiles and spectral signatures that were defining much better the crystal state of the chocolate samples. A two-component system explained the studied procedures, with a contribution S1 representing the highly crystalline state of the chocolate fats and S2 representative of the amorphous/less stable state. In general, we were able to track the transition from a less ordered to a highly ordered crystal state of the fats during the cooling stage of chocolate. After short storage, the chocolates had contributions of both S1 and S2 profiles. After long storage, the S1 crystalline form was the most dominant interpreted as the reorganization of the triglyceride acyl chains towards a thermodynamically favorable state. The latter was observed for all analyzed chocolates, implying that, regardless the tempering process or temper regime tested, the fats in chocolates reach the same high crystalline state after maturing.

Spontaneous fermentation is a process that depends on substrates' physical characteristics, crop variety, and postharvest practices; it induces variations in the metabolites that are responsible for the taste, aroma, and quality. Metabolomics makes it possible to detect key metabolites using chemometrics and makes it possible to establish patterns or identify biomarker behaviors under certain conditions at a given time. Therefore, sensitive and highly efficient analytical techniques allow for studying the metabolomic fingerprint changes during fermentation; which identify and quantify metabolites related to taste and aroma formation of an adequate processing time. This review shows that studying metabolomics in spontaneous fermentation permits the characterization of spontaneous fermentation in different stages. Also, it demonstrates the possibility of modulating the quality of cocoa by improving the spontaneous fermentation time (because of volatile aromatic compounds formation), thus standardizing the process to obtain attributes and quality that will later impact the chocolate quality.

The impact of cocoa lipid content on chocolate quality has been extensively described. Nevertheless, few studies have elucidated the cocoa lipid composition and their bioactive properties, focusing only on specific lipids. In the present study the lipidome of fine-flavor cocoa fermentation was analyzed using LC-MS-QTOF and a Machine Learning model to assess potential bioactivity was developed. Our results revealed that the cocoa lipidome, comprised mainly of fatty acyls and glycerophospholipids, remains stable during fine-flavor cocoa fermentations. Also, several Machine Learning algorithms were trained to explore potential biological activity among the identified lipids. We found that K-Nearest Neighbors had the best performance. This model was used to classify the identified lipids as bioactive or non-bioactive, nominating 28 molecules as potential bioactive lipids. None of these compounds have been previously reported as bioactive. Our work is the first untargeted lipidomic study and systematic effort to investigate potential bioactivity in fine-flavor cocoa lipids.

Green analysis is defined as the analysis of chemicals in a manner where sample extraction and analysis are performed with least amounts of steps, low hazardous materials, while maintaining efficiency in terms of analytes detection. Coffee and cocoa represent two of the most popular and valued beverages worldwide in addition to their several products i.e., cocoa butter, chocolates. This study presents a comprehensive overview of green methods used to evaluate cocoa and coffee seeds quality compared to other conventional techniques highlighting advantages and or limitations of each. Green techniques discussed in this review include solid phase microextraction, spectroscopic techniques i.e., infra-red (IR) spectroscopy and nuclear magnetic resonance (NMR) besides, e-tongue and e-nose for detection of flavor. The employment of multivariate data analysis in data interpretation is also highlighted in the context of identifying key components pertinent to specific variety, processing method, and or geographical origin.

Obesity causes inflammation which may lead to development of co-morbidities like cardiovascular diseases. Cocoa is a popular food ingredient that has been shown to mitigate obesity and inflammation in preclinical models. Cocoa typically undergoes fermentation and roasting prior to consumption, which can affect the polyphenol content in cocoa. The aim of this study was to compare the effect of fermentation and roasting protocols on the ability of cocoa to mitigate obesity, gut barrier dysfunction, and chronic inflammation in high fat (HF)-fed, obese C57BL/6J mice. We found that treatment of mice with 80 mg/g dietary cocoa powder for 8 weeks reduced rate of body weight gain in both male and female mice (46-57%), regardless of fermentation and roasting protocol. Colonic length was increased (11-24%) and gut permeability was reduced (48-79%) by cocoa supplementation. Analysis of the cecal microbiome showed that cocoa, regardless of fermentation and roasting protocol, reduced the ratio of Firmicutes to Bacteroidetes. Multivariate statistical analysis of markers of inflammation and body weight data showed sex differences in the effect of both the HF diet as well as cocoa supplementation. Based on this data there was strong protective efficacy from cocoa supplementation especially for the more processed cocoa samples. Overall, this study shows that anti-obesity and anti-inflammatory efficacy of cocoa is resilient to changes in polyphenol content and composition induced by fermentation or roasting. Further, this study shows that although cocoa has beneficial effects in both males and females, there are significant sex differences.

With an aim to prepare the healthier functional chocolate, olive oil-based diacylglycerol plastic fat (ODAGP) was prepared by mixing olive oil-based diacylglycerol stearin (O-DAGS) and olive oil-based diacylglycerol olein (O-DAGO) as confectionery fat in compound chocolate. We reported the physicochemical properties of ODAGP and ODAGP-CB blends, and then evaluated their application potential in compound chocolate based on sensory, blooming property and polymorphic transition. ODAGP (40% O-DAGS) showed a wide plastic range (7.1-45.2%) and high component compatibility. The crystal properties results indicated that ODAGP mainly existed in stable β-forms (β

The aim was to develop a fluorescent multiplex array for simultaneously measuring regulated food allergens using specific allergen protein molecules from peanut, tree nut, cow's milk, egg, soy, fish, shellfish, sesame, mustard and celery. Microspheres coupled to specific monoclonal antibodies were used for allergen detection, with purified allergens as reference standards.Standard curves for 17 allergens covered a 5-log dynamic range. Intra- and inter-assay acceptance criteria were within 70-130% recovery and a CV of ≤15%. Food reference materials contained high levels of their respective major allergens (2000-175,000 µg/g), Similar high allergen levels were found in 10 selected foods analysed using a 9-plex array. Egg, milk, peanut, hazelnut and walnut allergens were detectable in chocolate bars with incurred allergens at 3, 10, 30, and 100 ppm. The multiplex array is an efficient tool for measuring specific food allergens, with applications for risk assessment and standardization of therapeutic products for food allergy.

This study aimed to evaluate the volatile compounds of chocolates made of Brazilian cocoas and statistically track them according to the products' sensorial profile in order to relate them to consumers' acceptance by preference map methodology. The intensity of the chocolate, acidity, woody, smoked, green, floral, burned, musty, and cocoa notes from chocolates produced with cocoa from different Brazilian states were analyzed by a trained panel and by 128 consumers. Samples from Côte d'Ivoire, which is known for its high-quality chocolate, were evaluated for comparison. Solid-phase microextraction headspace sampling/gas chromatography-mass spectrometry was employed to evaluate the samples' volatile compounds. One hundred volatile compounds were identified within the samples. The results from the preference maps showed that the maximum preference was found for chocolate made of cocoa from Rondônia, Bahia, and Espírito Santo and Côte d'Ivoire and organic samples from Pará. The ideal sample point was characterized by intense chocolate, floral, and woody notes and mild green and burned notes. The presence of furfural, 3-methyl butanal, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, methyl pyrazine, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, and tetramethyl pyrazine were shown to be important for consumer acceptance in the ideal product, whereas the presence of (Z)-2-heptenal and 2-pentyl furan may increase consumer rejection. 2,3-Methyl pyrazine, methyl pyrazine, and 2,3-butanediol, which are important volatile compounds previously reported in the literature, were statistically tracked to both positive and negative sample attributes and must be better explored concerning consumers' acceptance of chocolates.

In this work, a facile ratiometric electrochemical aptasensor was developed towards sensitive and selective detection of vanillin, based on Ketjen black/ferrocene dual-doped zeolite-like MOFs (Fc-KB/ZIF-8) and electrodeposited gold nanoparticles (AuNPs) coupling with DNA aptamer. Fc-KB/ZIF-8 composites were prepared via one-pot solvothermal reaction and drop-coated on glassy carbon electrode (GCE) surface to form Fc-KB/ZIF-8@GCE. AuNPs were in-situ electro-deposited on the modified GCE. 5′-SH terminated aptamer of vanillin was combined with AuNPs via Au-S coupling to form aptamer-AuNPs/Fc-KB/ZIF-8@GCE as a new sensing platform. Under optimal conditions, electrochemical (square wave voltammetry) curves of this sensing platform were measured in electrolyte solutions containing vanillin. With increase of vanillin concentration (Cvan), vanillin had an increased peak current intensity (Ivan, as response signal). Fc doped into ZIF-8 had slight changes in its peak current intensity (IFc, as reference signal). There is a well plotting linear relationship between Ivan/IFc and the logarithm of Cvan ranging from 10 nM to 0.2 mM, with a low limit of detection of 3 nM. The aptamer-AuNPs/Fc-KB/ZIF-8@GCE was applied as a ratiometric electrochemical aptasensor of vanillin. This aptasensor had sensitive and selective electrochemical signal responses on vanillin, over potential interferents. This aptasensor enabled vanillin detection in real food samples, showing high detection performance. Experimental results testified that this aptasensor had high reliability and practicability for vanillin determination in real samples.

Bone loss resulting in increased risk for osteoporosis is a major health issue worldwide. Chocolate is a rich source of antioxidant and antiinflammatory flavonoids and dietary minerals with the potential to benefit bone health. However, other chocolate constituents such as cocoa butter, sugar, and methylxanthines may be detrimental to bone. Human studies investigating the role of chocolate consumption on serum bone markers and bone mineral density (BMD) have been inconsistent. A contributing factor is likely the different composition and thereby the nutrient and bioactive content among chocolate types. White and milk chocolate are high in sugar and low in flavonoids and most minerals. Dark chocolate (45-85% cocoa solids) is high in flavonoids, most minerals, and low in sugar with ≥70% cocoa solids resulting in higher fat and methylxanthine content. The aim of this review was to examine the relationship between chocolate consumption and its constiuents, including flavonoid content, on bone health and osteoporosis risk. Studies showed postmenopausal women had no bone effects at moderate chocolate intakes, whereas adolescents consuming chocolate had greater longitudinal bone growth. Based on flavonoid and mineral content, unsweetened cocoa powder appeared to be the best option followed by dark chocolate with higher cocoa content in terms of supporting and preserving bone health. Determining dietary recommendations for chocolate consumption relative to bone health is important because of the growing popularity of chocolate, particularly dark chocolate, and an expected increase in consumption owing to suggestions of health benefits against various degenerative diseases.

The detection of 3- and 2-MCPD ester and glycidyl ester was transformed from selected ion monitoring (SIM) mode to multiple reaction monitoring (MRM) mode by gas chromatography triple quadrupole spectrometry. The derivatization process was adapted from AOCS method Cd 29a-13. The results showed that the coefficient of determination (R2) of all detected compounds obtained from both detection mode was comparable, which falls between 0.997 and 0.999. The limit of detection and quantification (LOD and LOQ) were improved in MRM mode as compared to SIM mode. In MRM mode, the LOD of 3- and 2-MCPD ester was achieved 0.01 mg/kg while the LOQ was 0.05 mg/kg. Besides, LOD and LOQ of glycidyl ester were 0.024 and 0.06 mg/kg respectively. A blank spiked with MCPD esters (0.03, 0.10 and 0.50 mg/kg) and GE (0.06, 0.24 and 1.20 mg/kg) were chosen for repeatability and recovery tests. MRM mode showed better repeatability in area ratio and recovery with relative standard deviation (RSD %)

The aim of this study was to evaluate and explore the benefits of long-term dark chocolate intake in young, healthy adults by measuring cardiovascular function.A randomized study was conducted with 30 healthy participants ages 18 to 27 y. Half of the participants ingested a 20-g dose of lower cocoa chocolate (LCC; ∼55%; 12.6 ± 1.4 mg equivalent of epicatechin/g) and the others ingested a daily dose of 20 g of higher cocoa chocolate (HCC; ∼90%; 18.2 ± 2.6 mg equivalent of epicatechin/g). A baseline evaluation was performed before the participants started ingesting the assigned chocolate for a 30-d period, after which a final evaluation was performed. Each evaluation included heart ultrasonography, carotid-femoral pulse wave velocity (PWV) and carotid pulse wave analysis, flow-mediated slowing (FMS), and an analysis of the ventricular-arterial coupling (VAC), which reflects the matching between the aorta and the left ventricle (ratio of arterial elastance to left ventricle elastance).The baseline evaluation presented similar values within normal range in both groups. The positive vascular effects were overall more distinct in the group eating the HCC. No structural modifications on the heart were found after the intervention, notwithstanding cardiac function was improved on certain functional parameters in the HCC group only. A statistically significant improvement was depicted over the brachial and central systolic and pulse pressures in the HCC group, and a trend for improvement in the reflected waves component (Aix) and the FMS was also observed in the HCC, but not in the LCC group. VAC parameters were similar at baseline between groups, but showed a significant improvement in the HCC group after intervention, increasing from 0.674 to 0.719 (P = 0.004), so that the post-intervention VAC was significantly higher in the HCC group than in the LCC group (P0.05). In addition, significant variation was observed in both groups regarding arterial and left ventricle elastances, stroke work, and potential energy, with greater mean differences identified in the HCC group.This study demonstrated that regular consumption of HCC has beneficial effects on the cardiovascular system in young, healthy adults, improving vascular function by reducing central brachial artery pressures and promoting vascular relaxation, and thus enhancing the matching of the arterial system with the left ventricle.

Cocoa quality depends on several parameters, such as cocoa variety, environmental growth conditions, cultivation technique, and post-harvest treatments applied to coca beans. In this work, we studied the impact of cocoa post-harvest processing on both microbial communities structure and volatile composition. Cocoa beans samples were fermented in wooden boxes in Ivory Coast at different time intervals with turning and without turning, and derived from pods stored for two different duration times. Cocoa beans were analyzed using a molecular fingerprinting method (PCR-DGGE) in order to detect variations in microbial communities' structure; this global analysis was coupled to SPME-GC-MS for assessing cocoa volatile profiles. The results showed that the main parameter that influenced microbial communities structure was fermentation time, followed by turning, whereas, pods storage duration had a minor impact. Similar results were obtained for aromatic profile, except for pods storage duration that significantly affected volatile compound production. Global statistical analysis using Canonical Correspondence Analysis (CCA), showed the relationship between microbial communities and volatile composition. Furthermore, this study allowed the identification of discriminating microbial and chemical markers of cocoa post-harvest processing.

Food additives can be used to enhance processability and/or nutritional properties of food. In this study, two type of additives, magnesium stearate (Mg-ST) powder (as a processing aid) and plant sterol (PS) powder (as a processing and nutritional aids) were added into grated chocolate in order to enhance its flowability during auger type extrusion-based 3D printing. The thermal and rheology data showed that the melting peak of chocolate with additives ranged from 31.4 ± 0.8 °C to 32.1 ± 0.1 °C and a rapid reduction in viscosity began between 31.1 ± 0.7 °C and 33.3 ± 0.2 °C. The addition of additives showed no significant difference (p > .05) in melting peak (Tp) temperature of control chocolate samples and 3D printed chocolate samples suggesting that Mg-ST and PS do not affect the melting behaviour of chocolate. However, the yield stress values of chocolates containing additives were relatively higher than that of control samples indicating the possible effect of particulates. The tribology curve did not represent the typical Stribeck curve with or without the addition of additives. The particles in the additive potentially influenced the lubrication behaviour of 3D printed chocolate as their addition increased the coefficient of friction of the chocolate samples reducing the slippage in auger extrusion.

The impact of pod storage (PS) and roasting temperature (RT) on the aroma profiles of dark chocolates were evaluated. Cocoa liquor samples comprised of ten different combinations of PS and RT, whilst keeping the roasting time fixed at 35 min. Additionally, commercial cocoa liquors from renowned origins (Ecuador, Madagascar, Venezuela, Vietnam, Ivory Coast and Ghana) were acquired for comparison. From these, 70% dark chocolates were produced under the same conditions after which they were subjected to headspace solid-phase microextraction-gas chromatography–mass spectrometry (HS-SPME-GC–MS) analysis. Although both PS and RT were found to influence the aroma volatile concentrations, the impact of RT over PS seemed to be greater. An agglomerative hierarchical clustering (AHC) of all chocolates on the basis of their aroma profiles revealed a similar impact as earlier observed, where major clustering of the chocolates was in accordance with the intensity of the roasting process applied. However, within each group, the dissimilarities owing to PS among the chocolates was clearly depicted. Comparatively, chocolates with low (100–120 °C), instead of moderate to high (135–160 °C) RT's, rather showed a low dissimilarity with those from the commercial cocoa liquors of the different origins. Although from the same beans, the diversity of aroma profiles of these chocolates as well as the similitude of some treatments to some chocolates from commercial grade cocoa liquors, unequivocally underscores the possibility for steering diverse distinct flavors from ‘bulk’ cocoa through PS and roasting, with beneficial implications, both from an application and an economic point of view.

The present study has exploited the rapidity of the analysis and the multi-elemental capability of the energy dispersive X- ray fluorescence (ED-XRF) technique for the mineral profile determination in cocoa powder. A fast, cheap and environmental sustainable method without reagent consumption or toxic waste generation has been proposed. The samples can be prepared in the form of pellets of 13 mm in diameter and 2–3 mm thickness. The different internal calibrations used by ED-XRF equipment did not provide accurate results when comparing the mineral profile with the concentration obtained by Inductively Couple Plasma Optical Emission Spectroscopy (ICP-OES) after microwave assisted digestion of samples. For direct ED-XRF analysis of the cocoa samples, an external calibration using as standards the cocoa samples diluted with sugar was prepared. The analytical parameters of Relative Standard Deviation and Limit of Detection for the determined elements are adequate to the concentration levels found in the samples.

This study aimed to evaluate the effect of the process time (5, 10, and 15 min) and flow rate (10, 20, and 30 mL/min) of cold plasma technology on physio-chemical characteristics (pH), bioactive compounds (DPPD, Total Phenolic Compounds, ACE-inhibitory activity values), fatty acid composition, and volatile compounds profile of chocolate milk drink. The mild (lower flow rate and process time) and more severe (higher flow rate and process time) conditions led to a reduction of the bioactive compounds (total phenolic compounds and ACE-inhibitory activity), changes in fatty acid composition (increased saturated fatty acid and decreased monounsaturated fatty acid and polyunsaturated fatty acid), less favorable health indices (higher atherogenic, thrombogenic and hypercholesterolemic saturated fatty acids and lower desired fatty acids), and lower number of volatile compounds. In contrast, in intermediate cold plasma conditions, an adequate concentration of bioactive compounds, fatty acid composition, and health indices, and increased number of volatile compounds (ketones, esters, and lactones) were observed. Overall, cold plasma technology has proven to be an interesting alternative to chocolate milk drinks, being of paramount importance the study of the cold plasma process parameters.

Tracing the geographical origin of chocolate is of increasing importance owing to the market growth of cocoa products of high quality and especially where value is derived from those products being of single origin. The NMR analysis of methanolic/aqueous extracts of dark chocolate samples from Peru, Venezuela and Madagascar is reported and 42 different chemical constituents are identified, quantified and analysed using multivariate techniques. This paper describes a simple non-destructive protocol, which look at the chemical profile for chocolate samples from these three geographical locations and demonstrates potential for assessing the provenance of chocolate products, which has implications in food quality, safety and authenticity.

BACKGROUND: Placing food further away from people decreases likelihood of consumption ("Proximity Effect"). However, it is unclear how proximity affects consumption when both healthier and less healthy foods are available and cognitive resource for self-control is limited. AIMS: To test the hypothesis that when both healthier (raisins) and less healthy (chocolate M&Ms) foods are available, placing less healthy food far, rather than near, increases the likelihood that healthier food is consumed. METHODS: General population participants (N = 248) were all put under cognitive load and randomised to one of four groups: 1. Raisins near (20 cm), M&Ms far (70 cm); 2. Both foods near; 3. M&Ms near, raisins far; 4. Both far. PRIMARY OUTCOME: proportions of participants consuming raisins and M&Ms, respectively. RESULTS: The results did not support the primary hypothesis: when healthier and less healthy foods were both available, placing M&Ms far, rather than near, did not increase likelihood of consuming raisins (OR = 1.54, p = .432). Regardless of the M&Ms proximity, likelihood of consuming raisins was unaffected by the raisins' proximity (62.9%(near) vs. 56.5%(far) OR = 0.61, p = .211). Likelihood of consuming M&Ms non-significantly decreased when they were far and raisins were near, and when both foods were far (OR = 2.83, p = .057). Likelihood of consuming M&Ms was affected by M&Ms proximity, being higher when near (68.3%) than far (55.6%), OR = 0.39, p = .015. Indices of cognitive load impact (higher vs lower) were unrelated to consumption of either food. CONCLUSIONS: Likelihood of consuming a healthier food was unaffected by its proximity and that of a less healthy food. By contrast, likelihood of consuming a less healthy food was influenced by its proximity and possibly by that of a healthier food. These effects need replication in studies designed to detect smaller effect sizes. TRIAL REGISTRATION: This study was registered online with ISRCTN (ISRCTN11740813).

Flavonoid consumption has reported health benefits such as reducing cardiovascular disease risk factors, improving endothelial function, and delaying age-related cognitive decline. However, there are little dietary intake data for Australians, which limit our ability to make dietary recommendations to increase intakes to a level where health benefits are seen. The aim of this cross-sectional study was to determine the intake of flavonoids, flavonoid classes, and flavonoid subclasses of 1183 Australians aged 39 to 65 years using a validated 215-item food frequency questionnaire. Based on limited global flavonoid intake data, flavanols are the major dietary flavonoid and are found predominantly in tea and cocoa. As Australians are large tea drinkers, we anticipated that flavanols would be the major flavonoid in the Australian diet. The flavonoid content of foods was determined using a combination of the United States Department of Agriculture Databases and the Phenol-Explorer Database. One-way analysis of variance was undertaken to examine differences between flavonoid intake between men and women. Total flavonoid intake was 626 ± 579 mg/d. Men and women consumed 566 ± 559 mg and 660 ± 588 mg of total flavonoids per day, respectively. Thearubigin accounted for 58% of the flavonoid intake. Women consumed more total flavonoids, thearubigins (both P .01), anthocyanidins (P .0001), flavan-3-ols, flavones, and flavonols (all P .05) than men, whereas men consumed more flavanones than women (P = .01). There was no difference between sexes for the consumption of isoflavones. The data indicated that flavan-3-ols, predominantly thearubigin from tea, were the main flavonoid consumed by Australians. This information contributes to population flavonoid intakes, which should be considered when exploring flavonoid and health relationships.

A novel way to fully utilize rambutan fruit and seed is to ferment peeled fruits followed by drying and roasting, and use the seeds to produce seed powder similar to that of cocoa powder. Hence, the objective of this study was to optimize the roasting time and temperature of rambutan fruit post-fermentation and drying, and to produce a cocoa-like powder product from the seeds. Parameters monitored during roasting were colour and total phenolic content, while seed powder obtained using optimized roasting conditions was analyzed for its physicochemical properties and toxicity. The latter was examined using the brine shrimp lethality assay. Results showed that the roasted seed powder possessed colour and key volatile compounds similar to that of cocoa powder. Besides, the brine shrimp lethality assay indicated that the roasted seed powder was non-toxic. Thus, the fruit, including its seed could be fully utilized and subsequently, wastage could be reduced.

Pharmaceutical compounding is a core activity in the preparation of patient-specific dosage forms. In the current study we aimed to investigate whether 3D printing could be employed for the preparation of pediatric-friendly personalized dosage forms that fulfil the acceptance criteria specified in the pharmacopoeias for conventional dosage forms. We then compared the 3D printed dosage forms with the same formulations prepared with mold-casting, a method frequently applied during pharmaceutical compounding. The molded dosage forms failed to pass most of the quality control tests, including the mass uniformity and content uniformity tests, as well as dose accuracy, contrary to the 3D printed, which not only passed all tests but also enabled precision overdose adjustment. Hence, 3D printing of chocolate-based dosage forms may effectively serve as an acceptable alternative method to mold casting in compounding patient-specific medication at the point-of-care.

This study developed an electrocolorimetric extraction technique as a simple, rapid, and green method for the simultaneous preconcentration and determination of Ni(II) in chocolate samples. The system was designed using an agarose gel (3% w/v) solution containing 10% v/v 80 mM dimethylglyoxime (DMG) and 10% v/v 0.03 M ammonia as colorimetric reagents to determine Ni(II) ions. When voltage was applied to the system, Ni(II) ions were extracted from the donor solution into the gel and formed Ni-DMG complexes with a pink color. Under the optimal conditions, a good linear range was obtained from 30 to 750 µg L

The bioavailability impact of serum lipids in compound chocolate products based on structured lipids was studied. Compound chocolate products containing fat with and without structured lipids were digested in vitro under simulated gastrointestinal lipolysis conditions and were studied in vivo in healthy C57BL/6J mice. The in vitro digestion results show that products containing structured lipids, milk compound chocolate filling and white compound coating, significantly reduced the release rate of Free Fatty Acids (FFA) and improved the caloric reduction between 12.49% and 13.71% compared to products without structured lipids, suggesting that FFA were not absorbed. Animal feeding studies revealed no adverse effects on the compound products intake; in fact, these products reduced total cholesterol, LDL-c, VLDL-c and triacylglycerols. The present work shows the relevance of developing functional compound chocolate as providing a potential healthy initiative through the biological effect of the bioactive ingredients incorporated.

The gold standard method to isolate and identify Taylorella equigenitalis, the contagious agent of equine metritis, is the culture method according to the World Organisation for Animal Health Terrestrial Manual. No selective T. equigenitalis chocolate agar medium has been developed since the 1980s and the existing media show limited performances due to the fastidious nature of T. equigenitalis and the presence of interfering bacteria in the genital tract of equines. Here, the growth rates of 6 T. equigenitalis strains and 7 non-T. equigenitalis strains were compared on Timoney's selective medium formulated with 5 different basal agars (Columbia, Eugon, Blood, Mueller-Hinton and Tryptose Blood) provided by 2 to 4 suppliers per basal agar. The impact of glucose and/or Vitox supplementation was also investigated. Overall, the performance of selective T. equigenitalis media could be improved by substituting Eugon or Columbia agar with Blood, Mueller-Hinton or Tryptose Blood agar. It is nevertheless essential to validate the basal agar/supplier pair using a panel of T. equigenitalis strains. Furthermore, our findings confirm the need to supplement the selective media with a mixture of amino acids, nucleotides, and organic, mineral and vitamin compounds, translated here by Vitox supplementation.

Cocoa fermentation is the key and most relevant process in the synthesis of aroma and flavor precursor molecules in dry beans or raw material for producing chocolate. Because this process occurs in an uncontrolled manner, the chemical and sensory quality of beans can vary and be negatively affected. One of the strategies for the standardization and improvement of the sensory quality of chocolate is the introduction of microbial starter cultures. Among these, yeasts involved in fermentation have been studied because of their pectinolytic and metabolic potential in the production of volatile compounds. This study was aimed at isolating and characterizing, both sensory and chemically, yeasts involved in cocoa fermentation that could be used as starter cultures from two agro-ecological regions for the cultivation of cocoa in Colombia. The microbiological analyses identified 22 species represented mostly by Saccharomyces cerevisiae, Wickerhamomyces anomalus and Pichia sp. The preliminary sensory analysis of eight of these species showed that Hanseniaspora thailandica and Pichia kluyveri presented sensory profiles characterized by high intensity levels of fruity notes, which could be ascribed to the production of ethyl acetate, isoamyl acetate, and 2-phenylethyl acetate.

Chocolate manufacturers often develop their own original blends of cocoa beans, which, after processing, will give an unique sensory properties to their chocolates. In recent years, there has been a trend in the production of an increasing amount of low-processed or unprocessed food. Therefore, we carried out the research in which we obtained and analyzed chocolates with different blends of cocoa liquors prepared from roasted and unroasted cocoa beans of Criollo and Forastero varieties.Studies have shown that viscosity and yield value increased and hardness of chocolates lowered with the increase content of cocoa liquor from unroasted beans. Chocolates with cocoa liquor from roasted beans were characterized by the largest ability to scavenge DPPH radicals. Antioxidant activity determined by the ORAC-FL method increased with increasing polyphenol content in the chocolates. Type of the cocoa liquor used induced minor changes in the polyphenol and fatty acid composition. Asparagine was determined at the highest concentration, followed by alanine, glutamine, and serine. The concentration of acetic acid was the higher the higher the content of unroasted beans. The increased content of cocoa liquor from unroasted beans caused the color of the chocolate to brighten, more red and yellow. All tested chocolates were characterized by good organoleptic properties and microbial stability.

International audience; Ganache is a common chocolate preparation used as a filling for chocolate bonbons. The aim of this study was to understand the links between ganache composition, microstructure and macroscopic behaviour. The study proved that hot ganache is an oil-in-water emulsion, which evolves upon cooling into a bicontinuous system. Dark and white model ganaches were manufactured to understand the roles of components such as sugar, water, fat and cocoa in ganache structure and texture. Water activity and consistency of hot ganache, as well as firmness of crystallised ganache were measured. Real ganaches, standard and split (i.e. destabilised), were also manufactured, and analysed by confocal laser scanning microscopy (CLSM). Results showed that the main parameters explaining ganache behaviour and texture were the quantity of free water in ganache, and the balance between the aqueous phase and the fat phase. Results also highlighted the importance of non-fat cocoa in ganache, as a texturing agent that can help stabilise (in the case of standard ganache) or destabilise ganache (in the case of split ganache).

Recently, a huge number of studies have confirmed the important role of chocolate polyphenols in human health, underlining its beneficial effects especially in the treatment of cardiovascular diseases. However, a thorough evaluation of chocolate phenolic profile is still lacking. This study aimed at a comprehensive characterization of dark chocolate phenolic profile, using non-targeted mass spectrometry identification. This approach allowed a tentative identification of 158 individual phenolic compounds: 67 were newly detected in dark chocolate, among these 38 were observed for the first time in chocolate as well as in cocoa beans or products. Ellagitannins, which have never been reported in cocoa or chocolate, represented about the 10% of the phenolic profile of dark chocolate. The enrichment of dark chocolate with Sakura green tea leaves or turmeric powder influenced and modified the phenolic profile, resulting in a phenolic concentration increase. In this way, this functional chocolate might maximize the beneficial effect of chocolate consumption, combining the positive health effects of chocolate, turmeric and green tea and, at the same time, reducing the amount of sugars and calories introduced with chocolate.

Agradecimentos: This work was financially supported by Sao Paulo Research Foundation - FAPESP (grant numbers 2009/53006-0, 2012/10871-6); and PIBITI - UNICAMP Abstract: Two formulations of dark chocolate were developed by adding cocoa butter stearin (CBSt) or sorbitan monostearate (SMS) and compared to a standard formulation in order to investigate fat bloom formation over time. Fat bloom was monitored by Whiteness Index (WI), melting behavior and polymorphism determinations, in bars stored during 90 days at 20 degrees C and under oscillating temperature between 20 and 32 degrees C. All samples stored at 20 degrees C did not develop fat bloom and the required beta(V) form was maintained. Under oscillating storage condition, samples with CBSt (6.0%, w/w) and SMS (0.15%, w/w) delayed the surface fat bloom formation by at least 45 and 15 days, respectively, compared to standard chocolate, observed visually and through WI increments. The beta(V) to beta(VI) polymorphic transition correlated well with the WI, and also with changes in DSC thermograms, confirming the higher effectiveness of specific triacylglycerol (mainly StOSt) in delaying bloom formation FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Cocoa beans (Theobroma cacao L.) are the raw material for chocolate production. Fermentation of the bean pulp by microorganisms is essential for developing the precursors of chocolate flavour. Currently, the cocoa fermentation is still conducted by an uncontrolled traditional process via a consortium of indigenous species of yeasts, lactic acid bacteria and acetic acid bacteria. Although the essential contribution of yeasts to the production of good quality beans and, typical chocolate character is generally agreed, the roles of lactic acid bacteria and acetic acid bacteria are less certain. The objective of this study was to investigate the contribution of LAB and AAB in cocoa bean fermentation by conducting small scale laboratory fermentations under aseptic conditions, inoculated with different groups of microorganisms previously isolated from spontaneous cocoa fermentations. The inoculation protocols were: (1) four yeasts Hanseniaspora guilliermondii, Pichia kudriavzevii, Kluyveromyces marxianus and Saccharomyces cerevisiae; (2) four yeasts plus the lactic acid bacteria Lactobacillus plantarum and Lactobacillus fermentum; (3) four yeasts plus the acetic acid bacteria Acetobacter pasteurianus and Gluconobacter frateuri and (4) four yeasts plus two lactic acid bacteria and two acetic acid bacteria. Only the inoculated species were detected in the microbiota of their respective fermentations. Beans from the inoculated fermentations showed no significant differences in colour, shell weights and concentrations of residual sugars, alcohols and esters (p0.05), but they were slightly different in contents of lactic acid and acetic acid (p0.05). All beans were fully brown and free of mould. Residual sugar levels were less than 2.6 mg/g while the shell contents and ethanol were in the range of 11-13.4% and 4.8-7 mg/g, respectively. Beans fermented in the presence of LAB contained higher levels of lactic acid (0.6-1.2 mg/g) whereas higher concentrations of acetic acid (1.8-2.2 mg/g) were detected in beans inoculated with AAB. Triangle and hedonic sensory evaluations of chocolates prepared from beans taken from the three fermentations showed no significant differences (p 0.05). It was concluded that the growth of lactic acid bacteria and acetic acid bacteria may not be essential for the fermentation of cocoa beans.

Roasting is an important cocoa processing step, but has been reported to reduce the polyphenol content in the beans. We investigated the impact of whole-bean roasting on the polyphenol content, aroma-related chemistry, and in vitro pancreatic lipase (PL) inhibitory activity of cocoa under a range of roasting conditions. Total phenolics, (−)-epicatechin, and proanthocyanidin (PAC) dimer – pentamer content was reduced by roasting. By contrast, roasting at 150 °C or greater increased the levels of catechin and PAC hexamers and heptamers. These compounds have greater PL inhibitory potency. Consistent with these changes in PAC composition and this previous data, we found that roasting at 170 °C time-dependently increased PL inhibitory activity. Cocoa aroma-related compounds increased with roasting above 100 °C, whereas deleterious sensory-related compounds formed at more severe temperatures. Our results indicate that cocoa roasting can be optimized to increase the content of larger PACs and anti-PL activity, while maintaining a favorable aroma profile.

This study aimed to investigate the interactions between insoluble fractions of different coffee infusions and major cocoa free antioxidants, catechin and epicatechin, as well as the interactions between different coffee infusions and dark chocolate. Espresso, filtered coffee, French press and Turkish coffee were used for this purpose. Antioxidant capacity (AC) measurements were performed by monitoring the percentage inhibition of 2,2-Diphenyl-1-picrylhydrazil (DPPH) radical. Multivariate approach was adopted for experimental design and data analysis steps. In dry basis, the AC values of infusions (mmol Trolox/kg) were ranged between 953 ± 2.6 and 1184 ± 11.3, while the AC values for their insoluble fractions were ranged between 45 ± 0.0 and 105–1.3. Interactions between the insoluble fractions of coffee infusions and catechins were synergistic for espresso and additive/antagonistic for the other infusions. Interactions between coffee infusions and chocolate were synergistic for French press and Turkish coffee and additive/antagonistic for the other infusions.

The existence of sex differences was studied in a rat model of operant self-administration of a chocolate-flavored beverage (CFB), which possesses strong reinforcing properties and is avidly consumed by rats. Whether these differences occurred concomitantly to changes in extracellular dopamine in the dialysate obtained from the nucleus accumbens, was assessed by intracerebral microdialysis. Male, ovariectomized and intact female rats showed similar self-administration profiles, with minor differences in both acquisition and maintenance phases. Intact females self-administered larger amounts of CFB, when expressed per body weight, than males and ovariectomized females, in spite of similar values of lever-responding, latency to the first lever-response and consumption efficiency (a measure of rat’s licking effectiveness) in males, ovariectomized and intact females and no difference in breakpoint value and number of lever-responses emerged when males, ovariectomized and intact females were exposed to a progressive ratio schedule of reinforcement. Intracerebral microdialysis revealed a slight but significant increase in dopamine activity in the shell of the nucleus accumbens of male rats when compared to intact female rats during CFB self-administration. The above differences may be caused by the hormonal (mainly estradiol) fluctuations that occur during the estrus cycle in intact females. Accordingly, in intact females CFB self-administration and dopamine activity were found to fluctuate across the estrus cycle, with lower parameters of CFB self-administration and lower dopamine activity in the Proestrus and Estrus phases vs. the Metestrus and Diestrus phases of the cycle.

Children appear to engage in emotional eating (i.e., eating in response to negative and positive emotions), but existing research has predominantly relied on parent-report and child-report, which may not necessarily reflect children's actual emotional eating behaviors. This study examined the effects of happiness and sadness on children's observed snack consumption and examined whether child characteristics (i.e., weight, gender, and age) interact with mood to predict snack consumption. To elicit mood, children (N = 91; Mages = 6.8 years; 48 boys) were randomly assigned to one of the three mood induction conditions (happy, sad, or neutral); children's snack consumption was observed and measured after mood induction. Findings showed that children in the sad condition consumed more energy from chocolate, followed by children in the happy condition, and then the neutral condition. However, the reverse pattern was observed for goldfish crackers: children in the neutral condition consumed more energy from this savory snack than children in the happy condition, followed by children in sad condition. Child weight status and gender did not interact with mood to predict snack consumption. Child age did interact with mood: older children consumed more chocolates in the sad condition compared to younger children. Child age was not related to snack consumption in the happy and neutral conditions. This study suggests that emotional eating in response to positive and negative emotions is evident during early childhood, but that this is behavior is developing during this period.

Objective To test the hypothesis that epicatechin doses of 1.6, 3.2 and 4.8 mg per kg of body weight in a non-alkalized cocoa beverage cause significant acute decreases in human appetite that increase with increasing dose. Methods We conducted a four-way randomized, crossover, placebo-controlled double-blinded human trial with 28 healthy, non-obese, young-adult male participants. Our primary measure of appetite was ad libitum pizza intake 150 min after beverage ingestion. We used linear mixed models analyses. Results Compared to our placebo, the beverages containing 1.6, 3.2 and 4.8 mg per kg of body weight caused mean decreases in pizza intake of 984.3 kJ (18.1%, p = .002), 588.0 kJ (10.8%, p = .06) and 623.2 kJ (11.4%, p = .049), respectively. There were no reported adverse side effects related to the ingested beverages. Conclusions In our randomized trial two doses of epicatechin larger than1.6 mg/kg did not cause larger acute decreases in pizza intake.