Long-term supplementation with anthocyanin-rich or -poor Rubus idaeus berries does not influence microvascular architecture nor cognitive outcome in the APP/PS-1 mouse model of Alzheimer's disease.

Disruption of microvascular architecture is a common pathogenic mechanism in the progression of Alzheimer's disease (AD). Given the anti-angiogenic activity of berry (poly)phenols, we investigated whether long-term feeding of Rubus idaeus (raspberries) could ameliorate cerebral microvascular pathology and improve cognition in the APP/PS-1 mouse model of AD. Male C57Bl/6J mice (50 wild type, 50 APP/PS-1) aged 4-months were fed for 24-weeks, with a normal diet enriched with either 100 mg/day glucose (control diet) or supplemented with glucose and freeze-dried anthocyanin-rich (red) or -poor (yellow) raspberries (100 mg/day) and assessed/sampled post intervention. Cerebral microvascular architecture of wild-type mice was characterised by regularly spaced capillaries with uniform diameters, unlike APP/PS-1 transgenic mice which showed dysregulated microvascular architecture. Long-term feeding of raspberries demonstrated limited modulation of microbiota and no substantive effect on microvascular architecture or cognition in either mice model although changes were evident in endogenous cerebral and plasmatic metabolites.

Excretion by subjects on a low (poly)phenol diet of phenolic gut microbiota catabolites sequestered in tissues or associated with catecholamines and surplus amino acids.

Phenolic catabolites excreted by fasting subjects with a functioning colon and ileostomists on a low (poly)phenol diet have been investigated. Urine was collected over a 12 h fasting period after adherence to a low (poly)phenol diet for 36 h. UHPLC-HR-MS quantified 77 phenolics. Some were present in the urine of both groups in similar trace amounts and others were excreted in higher amounts by participants with a colon indicating the involvement of the microbiota. Most were present in sub- or low-µmol amounts, but hippuric acid dominated accounting on average for 60% of the total for both volunteer categories indicating significant production from sources other than non-nutrient dietary (poly)phenols. The potential origins of the phenolics associated with the low (poly)phenol diet, include endogenous catecholamines, surplus tyrosine and phenylalanine, and washout of catabolites derived from pre-study intakes of non-nutrient dietary (poly)phenols.

Effect of In Vitro Gastrointestinal Digestion and Colonic Fermentation on the Stability of Polyphenols in Pistachio (Pistacia Vera L.).

The aim of this study was to evaluate the impact of in vitro gastrointestinal digestion and colonic fermentation on the polyphenol compounds from different varieties of pistachio by UHPLC-HRMS analysis. The total polyphenol content decreased significantly, mostly during oral (recoveries of 27 to 50%) and gastric digestion (recoveries of 10 to 18%), with no significant changes after the intestinal phase. After in vitro digestion, the hydroxybenzoic acids and the flavan-3-ols were the main compounds found in pistachio, with respective total polyphenol contents of 73 to 78% and 6 to 11%. More specifically, the main compounds determined after in vitro digestion were 3,4,5-trihydroxybenzoic acid, vanillic hexoside and epigallocatechin gallate. The colonic fermentation affected the total phenolic content of the six varieties studied, with a recovery range of 11 to 25% after 24 h of fecal incubation. A total of twelve catabolites were identified after fecal fermentation, the main compounds being the 3-(3'-hydroxyphenyl)propanoic, 3-(4'-hydroxyphenyl)propanoic, 3-(3',4'-dihydroxyphenyl)propanoic, 3-hydroxyphenylacetic acids and 3,4-dihydroxyphenyl-É£-valerolactone. Based on these data, a catabolic pathway for colonic microbial degradation of phenolic compounds is proposed. The catabolites identified at the end of the process are potentially responsible for the health properties attributed to pistachio consumption.

Acute effect of oat ß-glucan on the bioavailability of orange juice flavanones.

The impact of ß-glucan on the bioavailability of orange juice (OJ) flavanones was investigated in a randomised controlled trial. Volunteers consumed 500 mL of OJ without or with either 3 g (OB-3) or 6 g (OB-6) of ß-glucan. Urine samples, collected 12 h before and over a 0-24 h period post-supplementation, were analysed by high-performance liquid chromatography-high resolution mass spectrometry. The overall 0-24 h urinary excretion of the 17 flavanone metabolites identified and quantified in urine after OJ ingestion corresponded to 29.7 µmol, and 25.0 and 9.3 µmol, respectively, after OB-3 and OB-6 intake. This corresponds to 9.3, 7.9, and 2.9% recoveries of the 318 µmol of the ingested flavanones. The acute ingestion of OJ with 6 g, but not 3 g of ß-glucan led to a significant reduction (p < 0.05) in the excretion of flavanone metabolites compared with consumption of OJ alone.

Impact of Metallic Nanoparticles on In Vitro Culture, Phenolic Profile and Biological Activity of Two Mediterranean Lamiaceae Species: Lavandula viridis L'Hér and Thymus lotocephalus G. López and R. Morales.

Nanoparticles (NPs) recently emerged as new chemical elicitors acting as signaling agents affecting several processes in plant metabolism. The aim of this work was to investigate the impact of the addition of copper oxide (CuO), zinc oxide (ZnO) and iron oxide (Fe3O4) NPs (<100 nm) at different concentrations (1, 5 and 10 mg/L) to the culture media on several morphological, physiological and -biochemical parameters of in vitro shoot cultures of Lavandula viridis L'Hér and Thymus lotocephalus G. López and R. Morales (Lamiaceae), as well as on phenolic profile and bioactivity (antioxidant and enzyme inhibition capacities). Although some decreases in shoot number and length were observed in response to NPs, biomass production was not affected or was improved in both species. Most NPs treatments decreased total chlorophyll and carotenoid contents and increased malondialdehyde levels, an indicator of lipid peroxidation, in both species. HPLC-HR-MS analysis led to the identification of thirteen and twelve phenolic compounds, respectively, in L. viridis and T. lotocephalus extracts, being rosmarinic acid the major compound found in all the extracts. ZnO and Fe3O4 NPs induced an increase in total phenolic and rosmarinic acid contents in T. lotocephalus extracts. Additionally, some NPs treatments also increased antioxidant activity in extracts from this species and the opposite was observed for L. viridis. The capacity of the extracts to inhibit tyrosinase, acetylcholinesterase and butyrylcholinesterase enzymes was not considerably affected. Overall, NPs had a significant impact on different parameters of L. viridis and T. lotocephalus in vitro shoot cultures, although the results varied with the species and NPs type.

Catabolism of citrus flavanones by the probiotics Bifidobacterium longum and Lactobacillus rhamnosus.

PURPOSE: Orange juice (OJ) flavanones undergo limited absorption in the upper gastrointestinal tract and reach the colon where they are transformed by the microbiota prior to absorption. This study investigated the ability of two probiotic bacteria, Bifidobacterium longum R0175 and Lactobacillus rhamnosus subsp. Rhamnosus NCTC 10302 to catabolise OJ flavanones. METHODS: The bacteria were incubated with hesperetin-7-O-rutinoside, naringenin-7-O-rutinoside, hesperetin and naringenin, and the culture medium and intracellular cell extracts were collected at intervals over a 48 h of incubation period. The flavanones and their phenolic acid catabolites were identified and quantified by HPLC-HR-MS. RESULTS: Both probiotics were able to subject hesperetin to ring fission yielding 3-(3'-hydroxy-4'-methoxyphenyl)propionic acid which was subsequently demethylated producing 3-(3',4'-dihydroxyphenyl)propionic acid and then via successive dehydroxylations converted to 3-(3'-hydroxyphenyl)propionic acid and 3-(phenyl)propionic acid. Incubation of both bacteria with naringenin resulted in its conversion to 3-(4'-hydroxyphenyl)propionic acid which underwent dehydroxylation yielding 3-(phenyl)propionic acid. In addition, only L. rhamnosus exhibited rhamnosidase and glucosidase activity and unlike B. longum, which was able to convert hesperetin-7-O-rutinoside and naringenin-7-O-rutinoside to their respective aglycones. The aglycones were then subjected to ring fission and further catabolised in a similar manner to that described above. The flavanones and their catabolites were found in the culture medium but not accumulated in the bacterial cells. CONCLUSIONS: These findings demonstrate the enzymatic potential of single strains of bifidobacterium and lactobacillus which may be involved in the colonic catabolism of OJ flavanones in vivo.

Effect of olive cultivar on bioaccessibility and antioxidant activity of phenolic fraction of virgin olive oil.

AIM: This study aims to characterize the phenolic profile and antioxidant capacity of seven monovarietal virgin olive oils (VOOs) and evaluate their in vitro gastrointestinal stability. METHODS: 'Picual', 'Blanqueta', 'Sevillana', 'Habichuelero', and 'Chetoui' olive cultivars were selected for VOO extraction. The oils were subjected to in vitro digestion. The recovery index (RI) of phenolic compounds after each digestion step and the bioaccessibility index (BI) were evaluated. In addition, the antioxidant activity of the bioaccessible fraction (BF) of VOOs was determined by DPPH, ABTS, and ORAC assays, as well as by studying the intracellular reactive oxygen species in Caco-2 cells. RESULTS: Differences were found in the composition of phenolic compounds in VOOs depending on cultivars. During the digestive process, important losses of phenolic compounds were observed between the buccal and duodenal steps, unlike HTy and Ty, which presented increased recovery due to the hydrolysis of secoiridoid derivatives. Differences in the bioaccessibility of phenolic compounds were found between varieties of VOOs. 'Sevillana' VOO had the highest total bioaccessibility (36%), followed by the 'Picual' (19%), 'Chetoui' (17%), 'Habichuelero' (10%), and 'Blanqueta' (8%) varieties. The BF of all the varieties of VOO showed similar radical ABTS scavenging capacity, 'Chetoui', and 'Blanqueta'-BF having the highest radical DPPH scavenging capacity, and 'Habichuelero' and 'Picual'-BF showing protective effects against the peroxyl radical measured by ORACFL assay. All VOO-BFs presented decreases in ROS levels in Caco-2 cells. CONCLUSIONS: Our results suggest differences in the bioaccessibility of phenolics from diverse VOO varieties, which could lead to different biological properties. Therefore, this study represents a first step toward the development of novel dietary strategies focusing on the phenolic supplementation of different VOOs to preserve human health.

Quantification of Total Phenolic and Carotenoid Content in Blackberries (Rubus Fructicosus L.) Using Near Infrared Spectroscopy (NIRS) and Multivariate Analysis.

A rapid method to quantify the total phenolic content (TPC) and total carotenoid content (TCC) in blackberries using near infrared spectroscopy (NIRS) was carried out aiming to provide reductions in analysis time and cost for the food industry. A total of 106 samples were analysed using the Folin-Ciocalteu method for TPC and a method based on Ultraviolet-Visible Spectrometer for TCC. The average contents found for TPC and TCC were 24.27 mg·g-1 dw and 8.30 µg·g-1 dw, respectively. Modified partial least squares (MPLS) regression was used for obtaining the calibration models of these compounds. The RPD (ratio of the standard deviation of the reference data to the standard error of prediction (SEP)) values from external validation for both TPC and TCC were between 1.5 < RPDp < 2.5 and RER values (ratio of the range in the reference data to SEP) were 5.92 for TPC and 8.63 for TCC. These values showed that both equations were suitable for screening purposes. MPLS loading plots showed a high contribution of sugars, chlorophyll, lipids and cellulose in the modelling of prediction equations.

Application of visible/near-infrared reflectance spectroscopy for predicting internal and external quality in pepper.

BACKGROUND: The characterization of internal (°Brix, pH, malic acid, total phenolic compounds, ascorbic acid and total carotenoid content) and external (color, firmness and pericarp wall thickness) pepper quality is necessary to better understand its possible applications and increase consumer awareness of its benefits. The main aim of this work was to examine the feasibility of using visible/near-infrared reflectance spectroscopy (VIS-NIRS) to predict quality parameters in different pepper types. Commercially available spectrophotometers were evaluated for this purpose: a Polychromix Phazir spectrometer for intact raw pepper, and a scanning monochromator for freeze-dried pepper. RESULTS: The RPD values (ratio of the standard deviation of the reference data to the standard error of prediction) obtained from the external validation exceeded a value of 3 for chlorophyll a and total carotenoid content; values ranging between 2.5 < RPD < 3 for total phenolic compounds; between 1.5 < RPD <2.5 for °Brix, pH, color parameters a* and h* and chlorophyll b; and RPD values below 1.5 for fruit firmness, pericarp wall thickness, color parameters C*, b* and L*, vitamin C and malic acid content. CONCLUSION: The present work has led to the development of multi-type calibrations for pepper quality parameters in intact and freeze-dried peppers. The majority of NIRS equations obtained were suitable for screening purposes in pepper breeding programs. Components such as pigments (xanthophyll, carotenes and chlorophyll), glucides, lipids, cellulose and water were used by modified partial least-squares regression for modeling the predicting equations. © 2015 Society of Chemical Industry.

Seasonal and Morphology Effects on Bioactive Compounds, Antioxidant Capacity, and Sugars Profile of Black Carrot (Daucus carota ssp. sativus var. atrorubens Alef.).

Black carrot (Daucus carota ssp. sativus var. atrorubens Alef.) is widely recognized for its bioactive compounds and antioxidant properties. The black carrot of Cuevas Bajas (Málaga) is a local variety characterized by a black/purple core, which differs from other black carrot varieties. Therefore, this autochthonous variety was characterized according to the root size and the harvesting season by means of a study of its antioxidant capacity analyzed by three methods, its total carotenoids content, and its sugars and phenolic compounds profile by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-MS). A total of 20 polyphenolic compounds were quantified in 144 samples analyzed. The anthocyanidins group was observed to be the most abundant, followed by the hydroxycinnamic acids group. Moreover, pelargonidin 3-sambubioside was observed in black carrot for the first time. The medium-sized carrots presented the highest content of phenolic compounds, largely due to their significantly higher anthocyanidins content. Comparatively, the small carrots showed a higher content of simple sugars than the large ones. Regarding the influence of season, significantly higher quantities of glucose and fructose were observed in the late-season carrots, while sucrose was the main sugar in early-season samples. No significant differences were observed in the total carotenoid content of black carrot.

Colon-available mango (poly)phenols exhibit mitigating effects on the intestinal barrier function in human intestinal cell monolayers under inflammatory conditions.

This study investigated the impact of in vivo available colon-mango (poly)phenols on stress-induced impairment of intestinal barrier function. Caco-2/HT29-MTX cells were incubated with six extracts of ileal fluid collected pre- and 4-8 h post-mango consumption before being subjected to inflammatory stress. (Poly)phenols in ileal fluids were analysed by UHPLC-HR-MS. Epithelial barrier function was monitored by measurement of trans-epithelial electrical resistance (TEER) and the production of selected inflammatory markers (interleukin-8 (IL-8) and nitric oxide (NO)) and the major mucin of the mucosal layer (MUC2). Post-mango intake ileal fluids contained principally benzoic acids, hydroxybenzenes and galloyl derivatives. There was a high interindividual variability in the levels of these compounds, which was reflected by the degree of variability in the protective effects of individual ileal extracts on inflammatory changes in the treated cell cultures. The 24 h treatment with non-cytotoxic doses of extracts of 4-8 h post-mango intake ileal fluid significantly reduced the TEER decrease in monolayers treated with the inflammatory cytomix. This effect was not associated with changes in IL-8 expression and secretion or claudine-7 expression. The mango derived-ileal fluid extract (IFE) also mitigated cytomix-dependent nitrite secretion, as a proxy of NO production, and the MUC2 reduction observed upon the inflammatory challenge. These insights shed light on the potential protective effect of mango (poly)phenols on the intestinal barrier exposed to inflammatory conditions.

Is the Intrasexual Competition in Male Red Deer Reflected in the Ratio of Stable Isotopes of Carbon and Nitrogen in Faeces?

Isotopic analysis of carbon and nitrogen in faeces is a reliable methodology for studying ecology in wildlife. Here, we tested this technique to detect variations in carbon and nitrogen isotopic ratios (δ13C and δ15N) in two different intrasexual competition scenarios of male Iberian red deer (Cervus elaphus hispanicus) using faeces of individuals collected during hunting actions in South-eastern Spain. The carbon isotopic ratio (δ13C) was not found to be significant, likely due to similar diet composition in all individuals. However, the nitrogen isotopic ratio (δ15N) was found to be lower in populations where sexual competition between males during the rut was higher compared to low-competition populations. Therefore, this study suggests a different use of proteins by an individual male red deer depending on the sexually competitive context in which he lives. Although further research is needed, these results show the potential of isotopic analysis as a tool for studying individual and populational variations in the level of intrasexual competition, with implications in evolutionary ecology and population management.

Urinary excretion of organosulfur compounds after acute ingestion of black onion.

Onion (Allium cepa L.) and its newly derived product "black onion" are characterised by the presence of compounds with potential bioactivity, particularly organosulfur compounds (OSCs). However, little is known about the metabolism, distribution, and excretion of these compounds as they pass through the gastrointestinal tract. This study monitored healthy subjects after an acute intake of black onion and analysed the excretion of OSCs using UHPLC-HRMS. A total of 31 OSCs were detected in urine after the acute ingestion of black onion, the main components being S-methyl-L-cysteine sulfoxide (methiin) (13.6 ± 3.9 µmol), isoalliin (12.4 ± 4.7 µmol) and S-propyl-L-cysteine (deoxypropiin) (3.1 ± 0.7 µmol). Moreover, N-acetylated metabolites of the major OSCs detected in black onion, namely, N-acetyl-S-(1-propenyl)-L-cysteine sulfoxide (NAS1PCS) and N-acetyl-S-(1-propenyl)-L-cysteine (NAS1PC), were found in urine after black onion consumption. The N-acetylation reaction takes place in the kidneys and liver, and metabolism pathways are proposed to explain the excretion of OSCs in urine. The basis of the identification of OSCs as urinary metabolites after black onion consumption is described for the first time and provides the basis for further research.

Bioavailability of Organosulfur Compounds after the Ingestion of Black Garlic by Healthy Humans.

The consumption of black garlic has been related to a decreased risk of many human diseases due to the presence of phytochemicals such as organosulfur compounds (OSCs). However, information on the metabolization of these compounds in humans is limited. By means of ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), this study aims to determine the OSCs and their metabolites excreted in urine 24 h after an acute intake of 20 g of black garlic by healthy humans. Thirty-three OSCs were identified and quantified, methiin (17,954 ± 6040 nmol), isoalliin (15,001 ± 9241 nmol), S-(2-carboxypropyl)-L-cysteine (8804 ± 7220 nmol) and S-propyl-L-cysteine (deoxypropiin) (7035 ± 1392 nmol) being the main ones. Also detected were the metabolites N-acetyl-S-allyl-L-cysteine (NASAC), N-acetyl-S-allyl-L-cysteine sulfoxide (NASACS) and N-acetyl-S-(2-carboxypropyl)-L-cysteine (NACPC), derived from S-allyl-L-cysteine (SAC), alliin and S-(2-carboxypropyl)-L-cysteine, respectively. These compounds are potentially N-acetylated in the liver and kidney. The total excretion of OSCs 24 h after the ingestion of black garlic was 64,312 ± 26,584 nmol. A tentative metabolic pathway has been proposed for OSCs in humans.

Bioavailability of orange juice (poly)phenols: ß-glucan-rich oat bran decreases urinary excretion of flavanone phase II metabolites and enhances excretion of microbiota-derived phenolic catabolites.

The impact of ß-glucan-rich oat bran on the bioavailability of orange juice (OJ) flavanones was investigated. Volunteers consumed 500 mL of OJ with and without 22 g of oat bran containing 6 g of ß-glucan (OB-6). Urine collected 12 h prior to and over a 0-24 h period post-supplementation was analysed by UHPLC-HRMS. Sixteen flavanone metabolites and thirty-nine colon-derived phenolic catabolites were identified and quantified. The major compounds were hesperetin-3'-glucuronide, along with hippuric acids and the C6-C3 phenolic acids 3-(3'-hydroxy-4'-methoxyphenyl)hydracrylic acid and 3-(4'-hydroxy-3'-methoxyphenyl)propanoic acid. A marked reduction in the 0-24 h excretion of flavanone metabolites from 29.7 µmol (9.3% recovery) to 9.3 µmol (2.9% recovery), occurred following consumption of OB-6 compared to OJ. This appeared not to be an effect of fiber on the rate of transport in the upper gut. After consumption of OJ there was a 163 ± 15 µmol excretion of colon-derived phenolic catabolites, equivalent to 43% of (poly)phenol intake and following OB-6 intake there was a further significant 30% increase. The ß-oat bran in OB-6 contained 5.8 µmol of free and 52 µmol of bound phenolic derivatives compared to 371 µmol of OJ (poly)phenols. The elevated excretion of phenolics after OB-6 consumption appears not to be due to bound phenolics in the bran, rather it is consequence, principally, of a bran-mediated increase in the quantities of flavanones passing from the upper to the lower bowel where they were subjected to microbiota-mediated catabolism. CLINICAL TRIAL REGISTRATION NUMBER: This trial was registered at clinicaltrials.gov as NCT04867655.

UHPLC-HRMS Spectrometric Analysis: Method Validation and Plasma and Urinary Metabolite Identification after Mango Pulp Intake.

After an acute intake of 300 g of mango purée by 10 subjects, 0 and 24 h urine and plasma samples were analyzed by high-performance liquid chromatography-high-resolution mass spectrometry. The method was first validated for 44 reference polyphenols in terms of linearity, specificity, limits of detection and quantification, intra-day and inter-day precision, recovery, and matrix effects in two biological matrices. After method validation, a total of 94 microbial-derived phenolic catabolites, including 15 cinnamic acids, 3 phenylhydracrylic acids, 14 phenylpropanoic acids, 12 phenylacetic acids, 28 benzoic acids, 2 mandelic acids, 15 hydroxybenzenes, and 5 hippuric acid derivatives, were identified or tentatively identified in urine and/or plasma. These results establish the value of the UHPLC-HRMS protocol and the use of authentic standards to obtain a detailed and accurate picture of mango polyphenol metabolites, together with their phase II conjugated metabolites, in human bioavailability studies.

Effect of Black Garlic Consumption on Endothelial Function and Lipid Profile: A Before-and-After Study in Hypercholesterolemic and Non-Hypercholesterolemic Subjects.

BACKGROUND: Black garlic is obtained from raw garlic (Allium sativum L.), by a fermentation process, under humidity and heat treatment, showing a high concentration of organosulfur compounds, which have been related to benefits in the prevention or delay of cardiovascular diseases (CVDs). The objective of the research was to evaluate whether long-term consumption of black garlic improves endothelial function and lipid profile in subjects with hypercholesterolemia. METHODS: Single center, controlled clinical trial with two branches: Hypercholesterolemia vs. Healthy condition. Sixty-two subjects of both sexes were distributed in two groups, the hypercholesterolemia group (n = 31) (total cholesterol (TC) range 200-300 mg/dL and low-density lipoprotein (LDL)-cholesterol range 135-175 mg/dL) and the healthy group (n = 31). The intervention consisted of the ingestion of 4 cloves of black garlic (12 g) daily for 12 weeks. RESULTS: significant increases in Apolipoprotein (Apo)A1 occurred in both groups: Hypercholesterolemia (Δ 11.8 mg/dL p < 0.001) vs Healthy (Δ 11.1 mg/dL p < 0.001). Besides, significant reductions for endothelial adhesion molecules monocyte chemoattractant protein-1 (MCP-1) (Δ -121.5 pg/mL p = 0.007 vs. Δ -56.3 pg/mL p = 0.015), intracellular adhesion molecule-1 (ICAM-1) (Δ -39.3 ng/mL p < 0.001 vs. Δ 63.5 ng/mL p < 0.001), and vascular cyto-adhesion molecule-1 (VCAM-1) (Δ -144.4 ng/mL p < 0.001 vs. Δ -83.4 ng/mL p = 0.061) were observed, for hypercholesterolemic and healthy subjects, respectively. CONCLUSIONS: These data show that black garlic consumption could improve some parameters related to endothelial function and lipid profile, which may have a favorable impact on the risk of CVDs, although more long-term studies are necessary to confirm.

Monitoring Changes in the Volatile Profile of Ecuadorian Cocoa during Different Steps in Traditional On-Farm Processing.

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

Effect of Drying Technique on the Volatile Content of Ecuadorian Bulk and Fine-Flavor Cocoa.

Cocoa (Theobroma cacao L.) is one of the most widely consumed edible seeds in the world affected by on-farm processing. This study investigated the effect of different drying techniques, namely oven drying (OD), sun drying (SD), and a modification of sun drying using black plastic sheeting (SBPD), on the volatile profile of fine-flavor and bulk cocoa varieties analyzed by HS-SPME-GC-MS. A total of sixty-four volatile compounds were identified in fresh and dried cocoa. As expected, the volatile profile was clearly modified after the drying step, showing strong differences among cocoa varieties, this factor and its interaction with the drying technique having greater influence according to the ANOVA simultaneous component analysis. A principal component analysis revealed a close relationship between the volatile content of bulk cocoa samples dried by the OD and SD techniques, whereas slight volatile modifications were perceived among fine-flavor samples dried using the three different techniques under study. Overall, the results provide the basis for the potential application of the simplest inexpensive SBPD technique in order to accelerate the drying process of sun drying and produce cocoa with similar (fine-flavor cocoa) or improved (bulk cocoa) aromatic quality to that formed using the traditional SD or the small-scale OD.

Exploring the potential links between gut microbiota composition and natural populations management in wild boar (Sus scrofa).

We surveyed wild boar (Sus scrofa) populations using 16S rRNA gene analysis of the gut microbiota in fresh faeces taken from 88 animals hunted in 16 hunting estates. The wild boar is a very convenient model system to explore how environmental factors including game management, food availability, disease prevalence, and behaviour may affect different biological components of wild individuals with potential implications in management and conservation. We tested the hypotheses that diet (according to stable carbon isotopes analyses), gender (i.e., animal behaviour studying males and females), and both health (analyses of serum samples to detect exposure to several diseases) and form statutes (i.e., thoracic circumference in adults) are reflected in changes in the intestinal microbiota. We focused on a gut functional biomarker index combining Oscillospiraceae and Ruminococcaceae vs. Enterobacteriaceae. We found that gender and the estate (population) were explanatory variables (c.a. 28% of the variance), albeit a high degree of overlapping among individuals was observed. The individuals with higher abundance of Enterobacteriaceae showed a gut microbiota with low diversity, mostly in males. Significant statistical differences for thoracic circumference were not found between males and females. Interestingly, the thoracic circumference was significantly and inversely related to the relative abundance of Enterobacteriaceae in males. Overall, we found that diet, gender, and form status were major factors that could be related to the composition and diversity of the gut microbiota. A high variability was observed in the biomarker index for populations with natural diet (rich in C3 plants). Although, we noticed a marginally significant negative trend between the index (higher abundance of Enterobacteriaceae) and the continuous feeding of C4 plants (i.e., supplementary maize) in the diet of males. This result suggests that continuous artificial feeding in hunting estates could be one of the factors negatively influencing the gut microbiota and the form status of wild boars that deserves further investigations.