Preliminary study of heart rate variability in Criollo horses for the elucidation of their neurophysiological characteristics of autonomic nerve function.

The Criollo is an Argentine horse breed with a calm temperament. Although its temperament is considered to be related to its neurophysiological characteristics, the details of this are unknown. Therefore, we analyzed the heart rate variability in Criollos as a preliminary study to deepen the neurophysiological understanding of their autonomic function. Electrocardiograms were recorded from Criollos and Thoroughbreds, and the power spectrum of heart rate variability was analyzed. Compared with Thoroughbreds, Criollos showed (i) a significantly higher high-frequency component, which is an index of parasympathetic nerve activity, and (ii) tendency toward a lower ratio of low- to high-frequency power, which is an index of the autonomic balance. These results revealed that parasympathetic nerves might be more active in Criollos compared with Thoroughbreds.

Characterisation of landrace 'criollo' maize silage from the highlands of Mexico in terms of starch content.

Maize silage is appreciated for dairy cows due to its high energy content. Mexico is a centre of origin for maize and small-scale dairy farmers use landrace 'criollo' maize silages to feed their herds. However, the characteristics of landrace 'criollo' maize silages in terms of starch content, which defines their nutritional value in terms of energy, are not known. The objective was to characterise criollo maize silages in terms of starch content. Seventy-seven silage samples from 2011 to 2015 were analysed for contents of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF), crude protein (CP), in vitro dry matter digestibility (IVDMD), and starch content. A factor analysis using principal component analysis as an extraction method was undertaken, and a hierarchical cluster analysis performed. Five factors affected the starch content of silages and five maize silage clusters identified. There was a positive association between IVDMD and metabolisable energy and between crude protein and acid detergent fibre; moreover, a positive relation was found between dry matter and maize starch contents. Significant differences were found (P < 0.001) for the contents of IVDMD, ME, ashes, OM, DM, and starch. The DM content of group 1 silage did not show any significant differences from other groups, except with group 2 silage, which had the lowest contents of DM and starch. Therefore, group 1 and 5 silage had the highest DM and starch contents, while group 3 silage had the highest IVDMD values and ME contents; finally, group 4 had the highest ash contents. Starch content in criollo maize silages was higher that reports for other maize races and is influenced by climate conditions and dry matter content.

Influence of Taxonomic and Functional Content of Microbial Communities on the Quality of Fermented Cocoa Pulp-Bean Mass.

Microbial metabolism drives changes in the physicochemical properties and, consequently, the sensory characteristics of fermented cocoa beans. In this context, information regarding the structure, function, and metabolic potential of microbial communities' present during cocoa pulp-bean mass fermentation is limited, especially concerning the formation of aromatic compounds. To bridge the gap, the metagenome of fermented cocoa pulp-bean mass (Criollo and Forastero) has been investigated using shotgun metagenomics coupled with physicochemical, microbiological, quality, and sensory analyses to explore the impact of microbial communities on the quality of fermented cocoa pulp-bean mass on one farm in one season and in one region under the same environmental conditions. Our findings showed that the metagenomic diversity in cocoa, the fermentation length, and the diversity and function of metagenome-assembled genomes (MAGs) greatly influence the resulting distinctive flavors. From the metabolic perspective, multiple indicators suggest that the heterolactic metabolism was more dominant in Criollo fermentations. KEGG genes were linked with the biosynthesis of acetic acid, ethanol, lactic acid, acetoin, and phenylacetaldehyde during Criollo and Forastero fermentations. MAGs belonging to Lactiplantibacillus plantarum, Limosilactobacillus reuteri, and Acetobacter pasteurianus were the most prevalent. Fermentation time and roasting are the most important determinants of cocoa quality, while the difference between the two varieties are relatively minor. The assessment of microbiological and chemical analysis is urgently needed for developing fermentation protocols according to regions, countries, and cocoa varieties to guarantee safety and desirable flavor development. IMPORTANCE Monitoring the composition, structure, functionalities, and metabolic potential encoded at the level of DNA of fermented cocoa pulp-bean mass metagenome is of great importance for food safety and quality implications.

Heat stress response in slick vs normal-haired Criollo Limonero heifers in a tropical environment.

To assess the effect of hair type on the heat stress response, 20 Criollo Limonero heifers with slick (n = 11) or normal hair (n = 9) were studied. Under a high temperature-humidity index (THI) environment, heat stress response was assessed through physiological variables that included respiration rate (RR), heart rate (HR), ruminal frequency (RMF), rectal temperature (RT), saliva pH (SPH), and lymphocyte count (LC) in the morning (5:00 AM, 27.4 °C, 64% relative humidity, THI = 77) and afternoon (1:00 PM, 34.5 °C, 70% relative humidity, THI = 88). A case-control study using a split plot design was used. Data were analyzed using ANOVA (PROC MIXED SAS 2010) and a statistical model comprising the fixed effects of hair length, sampling hour, interaction of hair length by sampling hour, and the random effect of animal nested within hair type on physiological variables associated with heat stress response. Sampling hour influenced (P < 0.0001) RR, RT, and (P < 0.003) SPH. Hair length influenced RR (P < 0.01) and RT (P < 0.04) and tended to influence LC (P < 0.07). The interaction of sampling hour by hair influenced RR (P < 0.04), RT (P < 0.0002), and both SPH and LC (P < 0.05). During afternoon hours, slick-haired heifers had lower values for RR (81 ± 4.2 vs 102 ± 4.7 bpm; P < 0.01), RT (39.5 ± 0.1 vs 40.3 ± 0.1 C°; P < 0.002), and LC (60 ± 3.2 vs 72.3 ± 3.6; P < 0.09) than normal-haired heifers. In normal-haired heifers, SPH increased during afternoon compared to morning-hours (8.66 ± 0.1 vs 9.11 ± 0.1; P < 0.04). It was concluded that slick-coated heifers exhibited an enhanced capability to cope with heat stress compared to normal-haired heifers likely due to an enhanced capacity for heat dissipation.

Seasonal variation of the estrous cycle length, corpus luteum area, and size of the pre-ovulatory follicle in Criollo Limonero heifers.

In order to determine whether seasonal variations may influence the estrous cycle length (ECL), corpus luteum size (CLS), maximum area of CL (MACL), day of cycle with maximum area of CL (DCMACL), and pre-ovulatory follicles size (PFS), ten Criollo Limonero heifers were subjected to daily ultrasound ovary scanning throughout their estrous cycles during three seasons: hot-dry (HD), hot-humid (HH), and wind-rain (WR). The effect of season on ECL, MACL, DCMACL, and PFS was analyzed with an ANOVA (PROC GLM, SAS), whereas, for the effect of season on CLS, an ANOVA with repeated measures (PROC MIXED, SAS) was used. Results showed no effect (P > 0.05) of season on ECL, MACL, and DCMACL. However, size of PFS was larger (P < 0.02) during the WR season and the CLS tended (P < 0.09) to be lower during the HH. In conclusion, the relative stability of ECL, MACL, DCMACL, PFS, and CLS measures suggests no major seasonal variations which could imply adaptation capability of Criollo Limonero cattle to the tropical environment.

Kappa-casein genotyping in tropical milking Criollo and its association to milk production and composition.

Caseins are abundant proteins in milk and found in four types (αS1, αS2, ß, and κ). There is substantial variation in the allelic and genotypic frequencies of the κ-casein gene in different cattle breeds, although the tropical milking Criollo (TMC) has not yet been investigated. The aim was to determine the allelic and genotypic frequencies in the κ-casein gene for two alleles (A and B) in TMC and further investigate its association to milk production and composition. A total of 180 TMC females were genotyped from blood samples. κ-Casein genotyping was performed using restriction fragment length polymorphism (RFLP) after polymerase chain reaction (PCR)-based amplification of genomic DNA. Allele frequencies were 0.39 for A-allele and 0.61 for B-allele (P < 0.05). Genotype frequencies were 0.09 (AA), 0.60 (AB), and 0.31 (BB) (P < 0.05). The κ-casein genotype in TMC cows did not affect milk yield or composition. In sum, the TMC has high frequencies of the B-allele and AB/BB genotypes, although there are no association of such genotypes and milk traits.

Genome-wide differential DNA methylation in tropically adapted Creole cattle and their Iberian ancestors.

Enhancing climate resilience and sustainable production for animals in harsh environments are important goals for the livestock industry given the predicted impacts of climate change. Rapid adaptation to extreme climatic conditions has already been imposed on livestock species, including those exported after Columbus's arrival in the Americas. We compared the methylomes of two Creole cattle breeds living in tropical environments with their putative Spanish ancestors to understand the epigenetic mechanisms underlying rapid adaptation of a domestic species to a new and more physiologically challenging environment. Reduced representation bisulfite sequencing was used to assess differences in methylation in Creole and Spanish samples and revealed 334 differentially methylated regions using high stringency parameters (P-value <0.01, ≥4 CpGs within a distance of 200 bp, mean methylation difference >25%) annotated to 263 unique features. Gene ontology analysis revealed candidate genes involved in tropical adaptation processes, including genes differentially hyper- or hypomethylated above 80% in Creole samples displaying biological functions related to immune response (IRF6, PTGDR, FAM19A5, PGLYRP1), nervous system (GBX2, NKX2-8, RPGR), energy management (BTD), heat resistance (CYB561) and skin and coat attributes (LGR6). Our results entail that major environmental changes imposed on Creole cattle have had an impact on their methylomes measurable today, which affects genes implicated in important pathways for adaptation. Although further work is needed, this first characterization of methylation patterns driven by profound environmental change provides a valuable pointer for the identification of biomarkers of resilience for improved cattle performance and welfare under predicted climatic change models.

The cacao Criollo genome v2.0: an improved version of the genome for genetic and functional genomic studies.

BACKGROUND: Theobroma cacao L., native to the Amazonian basin of South America, is an economically important fruit tree crop for tropical countries as a source of chocolate. The first draft genome of the species, from a Criollo cultivar, was published in 2011. Although a useful resource, some improvements are possible, including identifying misassemblies, reducing the number of scaffolds and gaps, and anchoring un-anchored sequences to the 10 chromosomes. METHODS: We used a NGS-based approach to significantly improve the assembly of the Belizian Criollo B97-61/B2 genome. We combined four Illumina large insert size mate paired libraries with 52x of Pacific Biosciences long reads to correct misassembled regions and reduced the number of scaffolds. We then used genotyping by sequencing (GBS) methods to increase the proportion of the assembly anchored to chromosomes. RESULTS: The scaffold number decreased from 4,792 in assembly V1 to 554 in V2 while the scaffold N50 size has increased from 0.47 Mb in V1 to 6.5 Mb in V2. A total of 96.7% of the assembly was anchored to the 10 chromosomes compared to 66.8% in the previous version. Unknown sites (Ns) were reduced from 10.8% to 5.7%. In addition, we updated the functional annotations and performed a new RefSeq structural annotation based on RNAseq evidence. CONCLUSION: Theobroma cacao Criollo genome version 2 will be a valuable resource for the investigation of complex traits at the genomic level and for future comparative genomics and genetics studies in cacao tree. New functional tools and annotations are available on the Cocoa Genome Hub ( http://cocoa-genome-hub.southgreen.fr ).

The legacy of Columbus in American horse populations assessed by microsatellite markers.

Criollo horse populations descend from horses brought from the Iberian Peninsula over the period of colonization (15th to 17th century). They are spread throughout the Americas and have potentially undergone genetic hybridization with other breeds in the recent past. In this study, 25 autosomal microsatellites were genotyped in 50 horse breeds representing Criollo populations from 12 American countries (27 breeds), breeds from the Iberian Peninsula (19), one breed each from France and Morocco and two cosmopolitan horse breeds (Thoroughbred and Arabian). The genetic relationships among breeds identified five clusters: Celtic; Iberian; North American with Thoroughbred influence; most Colombian breeds; and nearly all other Criollo breeds. The group of "all other Criollo breeds" had the closest genetic relationship with breeds originating from the Iberian Peninsula, specifically with the Celtic group. For the whole set of Criollo breeds analysed, the estimated genetic contribution from other breeds was approximately 50%, 30% and 20% for the Celtic, Iberian and Arab-Thoroughbred groups, respectively. The spatial distribution of genetic diversity indicates that hotspots of genetic diversity are observed in populations from Colombia, Ecuador, Brazil, Paraguay and western United States, possibly indicating points of arrival and dispersion of Criollo horses in the American continent. These results indicate that Criollo breeds share a common ancestry, but that each breed has its own identity.

Roasting conditions for preserving cocoa flavan-3-ol monomers and oligomers: interesting behaviour of Criollo clones.

BACKGROUND: Cocoa bean roasting is important for creating the typical chocolate aroma through Maillard reactions, but it is also a key step deleterious to the polyphenol content and profile. RESULTS: Compared with usual roasting at 150 °C, keeping the beans for 30 min at 120 °C or for 1 h at 90 °C proved much better for preventing strong degradation of native P1, P2 and P3 flavan-3-ols in cocoa (shown for Forastero, Trinitatio and Criollo cultivars). Surprisingly, Cuban, Mexican and Malagasy white-seeded beans behaved atypically when roasted for 30 min at 150 °C, releasing a pool of catechin. Enantiomeric chromatographic separation proved that this pool contained mainly (-)-catechin issued from (-)-epicatechin by epimerisation. As the (-)-epicatechin content remained relatively constant through Criollo bean roasting, flavan-3-ol monomers must have been regenerated from oligomers. This emergence of (-)-catechin in Criollo beans only, reported here for the first time, could be due to increased flavan-3-ol monomer stability in the absence of anthocyanidin-derived products. CONCLUSION: The degradation rate of flavan-3-ols through roasting is higher in cocoa beans containing anthocyani(di)ns. The liberation of a pool of (-)-catechin when submitted to roasting at 150 °C allows to distinguish white-seeded cultivars. © 2017 Society of Chemical Industry.

Relevance of Individual Data When Assessing the Gastrointestinal Nematode Infection Level, Nutritional and Productive Variables in a Tropical Farm Context: The Median Isn't the Message.

We evaluated the relationship between individual and herd GIN infection level, nutrition, production performance and anemia parameters in a tropical farm context. Fifty-four female goats were monitored to assess their body condition score (BCS, nutritional status indicator), live weight (LW) and LW gain (LWG, both used as production level indicators), FAMACHA© and hematocrit (HT, both used as anemia indicators). Goats browsed for 4 h in a tropical forest and received balanced feed and chopped grass. The eggs per gram of feces (EPG) indicated the GIN burden, with fecal samples obtained at 7:00 (AM) and 15:00 h (PM.) from each goat at six sampling points during the study. The variables and their relationship with GIN burdens were analyzed using Kruskall-Wallis, ANOVA and Friedman tests and Spearman correlations. The fecal samples obtained in the AM and PM can be equally representative of parasitic burdens (similar and highly correlated). However, the EPG of individual goats from periods of 30 days apart can be considered independent. The BCS and LWG varied between sampling times (p < 0.05), whereas EPG, LW and HT did not (p > 0.05). The GIN burden was negatively correlated with HT and BCS (-0.21, p = 0.01 for each one). The individual pattern of infection demonstrates the true impact of GINs on their hosts. Additionally, feeding and nutritional status may present important variations influencing the performance of the goats more than the impact of GINs under the farm conditions of the present study. However, GIN infection contributed to the variation in goat health and productivity in this tropical farm.

Reduction in the Cocoa Spontaneous and Starter Culture Fermentation Time Based on the Antioxidant Profile Characterization.

In current systems, the fermentation spontaneous process produces fermented beans of heterogeneous quality due to the fermentation time. This study demonstrated that the fermentation time should be reduced. For this purpose, the physicochemical parameters, antioxidant profile, and volatile compounds were characterized in two types of fermentation (spontaneous and starter culture) for 168 h in cocoa from three altitude levels. Multivariate analysis (cluster and PCA) was used to discriminate the fermentation stages. We found three stages in all fermentations, where the first two stages (0 h to 96 h) were characterized by a higher antioxidant potential of the cocoa bean and the presence of desirable volatile compounds such as acids, alcohols, aldehydes, ketones, and esters, which are precursors of cocoa aroma; however, prolonged fermentation times affected the antioxidant profile of the bean. In addition, the use of a starter culture facilitates the release of compounds in a shorter time (especially alcohols and esters). It is concluded that it is necessary to reduce the fermentation time under these conditions in the region of Amazonas.

Proximate Composition and Bioactive Compounds of Cocoa Bean Shells as a By-Product from Cocoa Industries in Indonesia.

Cocoa bean shell (CBS) is a by-product from cocoa processing which is abundant in Indonesia, one of the largest cocoa-producing countries. It has a great potential for being processed into food ingredients due to its comparable composition to cocoa nibs. The present study was conducted to identify the proximate composition and bioactive compounds in CBS produced at several cocoa industries in Indonesia utilizing different cocoa varieties (Criollo and Forastero) and processing techniques (fermented, non-fermented, pulp washing, and drying), which remain unknown. The results showed that the CBS derived from roasted Criollo cocoa pods in the Kendeng Lembu cocoa industry had ash and protein content of about 8.21% and 18.79%, respectively, which was higher than other industries. Additionally, the concentration of bioactive substances was higher here than it was elsewhere. This included total phenolic (136.2 mg GAE g-1) and theobromine (22.50 mg g-1). The lowest ash and protein concentration found in CBS was from Forastero cocoa pods, non-fermented like Sulawesi cocoa. These values were 6.48% and 15.70%, respectively. The concentration of theobromine (15.40 mg g-1) was also lower compared to other industries.

Chili Pepper Jojutla Morelos (Capsicum annuum L.), CJ-2018: A Variety Resistant to Bactericera cockerelli.

Chili pepper is a vegetable of worldwide economic and gastronomic importance. The psyllid, Bactericera cockerelli, is an economically important pest in this crop, causing considerable losses in its production. Currently, the application of insecticides is the main way to control B. cockerelli. However, the use of varieties resistant to this insect is a viable alternative for its control and management. In this work, the oviposition rate, development, and survival of B. cockerelli in two native varieties of chili were evaluated. Choice and non-choice trials showed that the B. cockerelli oviposition was reduced on CJ-2018 by 92.17 and 80.18%, respectively, compared to the control. In CM-334, the insect showed a behavior similar to the control in the non-choice test, while in the choice test it laid more eggs on CM-334 compared to the control. The development and survival assay showed that only 1.33% of the eggs managed to reach the adult stage on CJ-2018. In contrast, on CM-334 the survival of B. cockerelli was similar to the control. These results suggest that CJ-2018 presented a resistance based on antixenosis and antibiosis against B. cockerelli.

Use of molecular networking to identify 2,5-diketopiperazines in chocolates as potential markers of bean variety.

2,5-diketopiperazines are cyclic dipeptides found, among others, in chocolate. Although those compounds are contributing greatly to its pleasant bitterness, they can also be seen as interesting markers of cocoa beans processing. To evaluate the influence of bean variety and processing technology on the quantity of 2,5-diketopiperazines formed in chocolates, HPLC-MS/MS analyses were conducted, and a molecular network was built with the MS2 data. This approach eases the identification of 2,5-diketopiperazines within complex datasets and allows to visualize the chemical diversity of all samples. Using this methodology, 33 dark chocolates were analysed. 18 different diketopiperazine were identified and quantified. Among them, cyclo(L-ile-L-val), cyclo(L-leu-L-ile) and cyclo(L-phe-L-phe) were, to the best of our knowledge, detected for the first time in chocolate. The molecular network allows the clear visualization of differences between samples. The principal component analysis revealed the clustering of small batch chocolate samples according to bean variety, suggesting that bean genotype has a strong influence on the 2,5-diketopiperazines content of bean-to-bar chocolates, regardless of the degree of roasting or the technological process used by the small producers. The presence of two unique diastereoisomers in the classical chocolates bought in the supermarket indicates that the beans have probably undergone a more intense heat treatment. This study proposes the use of 2,5-diketopiperazines as potential markers of cocoa beans variety, as well as an indicator of post-harvest processing and processing technology, and highlights the potential of the molecular networks in the field of food and drink innovation as a promising tool to understand the complex chemistry of flavours.

Kinetics Crystallization and Polymorphism of Cocoa Butter throughout the Spontaneous Fermentation Process.

The spontaneous fermentation process of Criollo cocoa is studied for its importance in the development of chocolate aroma precursors. This research supports the importance of spontaneous fermentation, which was studied through the crystallization behavior and polymorphisms of cocoa butter (CB), the most abundant component of chocolate that is responsible for its quality physical properties. The k-means technique was used with the CB crystallization kinetics parameters to observe the division of the process during the first stage (day 0-3). The experimental crystallization time was 15.78 min and the second stage (day 4-7) was 17.88 min. The Avrami index (1.2-2.94) showed that the CB crystallizes in the form of a rod/needle/fiber or plate throughout the process. CB produced metastable crystals of polyforms ß1' and ß2'. Three days of fermentation are proposed to generate Criollo cocoa beans with acceptable CB crystallization times.

Inheritance of Yield Components and Morphological Traits in Avocado cv. Hass From "Criollo" "Elite Trees" via Half-Sib Seedling Rootstocks.

Grafting induces precocity and maintains clonal integrity in fruit tree crops. However, the complex rootstock × scion interaction often precludes understanding how the tree phenotype is shaped, limiting the potential to select optimum rootstocks. Therefore, it is necessary to assess (1) how seedling progenies inherit trait variation from elite 'plus trees', and (2) whether such family superiority may be transferred after grafting to the clonal scion. To bridge this gap, we quantified additive genetic parameters (i.e., narrow sense heritability-h 2, and genetic-estimated breeding values-GEBVs) across landraces, "criollo", "plus trees" of the super-food fruit tree crop avocado (Persea americana Mill.), and their open-pollinated (OP) half-sib seedling families. Specifically, we used a genomic best linear unbiased prediction (G-BLUP) model to merge phenotypic characterization of 17 morpho-agronomic traits with genetic screening of 13 highly polymorphic SSR markers in a diverse panel of 104 avocado "criollo" "plus trees." Estimated additive genetic parameters were validated at a 5-year-old common garden trial (i.e., provenance test), in which 22 OP half-sib seedlings from 82 elite "plus trees" served as rootstocks for the cv. Hass clone. Heritability (h 2) scores in the "criollo" "plus trees" ranged from 0.28 to 0.51. The highest h 2 values were observed for ribbed petiole and adaxial veins with 0.47 (CI 95%0.2-0.8) and 0.51 (CI 0.2-0.8), respectively. The h 2 scores for the agronomic traits ranged from 0.34 (CI 0.2-0.6) to 0.39 (CI 0.2-0.6) for seed weight, fruit weight, and total volume, respectively. When inspecting yield variation across 5-year-old grafted avocado cv. Hass trees with elite OP half-sib seedling rootstocks, the traits total number of fruits and fruits' weight, respectively, exhibited h 2 scores of 0.36 (± 0.23) and 0.11 (± 0.09). Our results indicate that elite "criollo" "plus trees" may serve as promissory donors of seedling rootstocks for avocado cv. Hass orchards due to the inheritance of their outstanding trait values. This reinforces the feasibility to leverage natural variation from "plus trees" via OP half-sib seedling rootstock families. By jointly estimating half-sib family effects and rootstock-mediated heritability, this study promises boosting seedling rootstock breeding programs, while better discerning the consequences of grafting in fruit tree crops.

Immune response of adult horses, pregnant mares and foals to an experimental vaccine with recombinant EMA-2 protein of Theileria equi.

Equine theileriosis, caused by the Theileria equi protozoan, is a disease of worldwide importance. T. equi expresses surface proteins, of which the EMA-2 protein is a promising antigen for vaccine use. The aim of this study was to evaluate the immune response of adult horses, pregnant mares, and foals to an experimental EMA-2 protein of recombinant T. equi vaccine. A total of 46 horses were used in this study for vaccine trials and challenges. Twelve geldings, 14 pregnant mares, and 14 foals were divided into vaccinated and control groups. Total serum specific anti-rEMA-2 IgG, IgG subclasses, and transcription of cytokines related to the immune response were evaluated. For the vaccine challenge, six six-month-old foals were divided into vaccinated and control groups. For the challenge, blood from a horse with theileriosis was transfused to the foals. Geldings and pregnant mares maintained anti-rEMA-2 IgG levels at 130 and 140 days after vaccination, respectively. The most-detected IgG subclasses in vaccinated were IgG3/5, IgG4/7, and IgG1. IL2, IL10, IL12, IL17, IFN-γ, and TNF-α were the most-transcribed cytokines in PBMCs of vaccinated horses stimulated with rEMA-2. Challenge with T. equi demonstrated that vaccinated foals had an increase of 33% in total IgG four days after blood transfusion, while control foals had no significant response, suggesting that vaccine antibodies may have recognized EMA-2 protein of the native T. equi antigen. T. equi recombinant EMA-2 was shown to be a promising vaccine antigen by inducing humoral and cellular immunity similar to that observed in natural parasite infections.

Theobromacacao Criollo var. Beans: Biological Properties and Chemical Profile.

Theobroma cacao provides precious products such as polyphenol-rich beans that are useful for nutraceutical purposes. The geographical area may influence the chemical composition of raw cocoa beans in terms of the polyphenols and biological qualities of the products. This work aimed to investigate the biological properties and the chemical composition of two different samples of Criollo var. cocoa raw beans coming from two areas (Indonesia; Peru). Beans underwent biphasic extraction obtaining lipophilic and hydroalcoholic extracts. The extracts were tested for antiradical, antimutagenic, and antigenotoxic effects. Cell viability inhibition toward breast, gastric/esophageal colorectal adenocarcinoma, and hepatoblastoma human cell lines was evaluated. Extracts were chemically investigated through UV-Vis spectroscopy and ultra-high-pressure liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QqTOF MS/MS). Results showed that the Indonesian bean hydroalcoholic extracts were able to scavenge 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cation radical better than the Peruvian hydroalcoholic extracts (ECs50: 72.63 vs. 322.20 µg/mL). Extracts showed antimutagenic and antigenotoxic activity. The viability inhibitory effect on breast and hepatic cancer cells was reached only for the Indonesian hydroalcoholic extracts at hundreds of µg/mL. Phenylpropenoyl-L-amino acids, hydroxycinnamoyl aminoacids conjugates, and procyanidin compounds were found mainly in the hydroalcoholic extracts, whereas fatty acids and lyso-phospholipids were found mainly in lipophilic fractions. Fatty acid and (epi)catechins appeared to be affected by different environmental conditions of the geographical areas.

Rheological, bioactive properties and sensory preferences of dark chocolates with partial incorporation of Sacha Inchi (Plukenetia volubilis L.) oil.

We studied the effect of substituting partially, cocoa butter (CB) with Sacha Inchi (Plukenetia volubilis L.) oil (SIO) on rheology, bioactive properties, and sensory preferences in potentially functional chocolate. For this 70% dark chocolates were prepared and the CB was substituted with 1.5%, 3%, and 4.5% of SIO. Hardness and viscosity of the SIO-chocolates were significantly reduced compared to the control (5451 ± 658 g; 17.01 ± 0.94 Pa s, respectively). Total phenolic content remained constant while the antioxidant capacity increased up to IC50 of 2.48 ± 0.10 as the content of SIO increased. The Casson yield stress and Casson plastic viscosity decreased as the amount of SIO increased. Chocolates with 4.5% SIO had a similar color, better glossiness, preferable snap attributes, and were more accepted (7.50 ± 0.08) compared to the control (p < 0.05), measured with a hedonic scale. Then, SIO can improve the bioactive properties of dark chocolates obtaining a potentially functional food with acceptable physicochemical characteristics. SIO can be considered as a new cocoa butter equivalent.