Effects of Commercial Apple Varieties on Human Gut Microbiota Composition and Metabolic Output Using an In Vitro Colonic Model.
ABSTRACT: Apples are a rich source of polyphenols and fiber. A major proportion of apple polyphenols escape absorption in the small intestine and together with non-digestible polysaccharides reach the colon, where they can serve as substrates for bacterial fermentation. Animal studies suggest a synergistic interaction between apple polyphenols and the soluble fiber pectin; however, the effects of whole apples on human gut microbiota are less extensively studied. Three commercial apple varieties-Renetta Canada, Golden Delicious and Pink Lady-were digested and fermented in vitro using a batch culture colonic model (pH 5.5-6.0, 37 °C) inoculated with feces from three healthy donors. Inulin and cellulose were used as a readily and a poorly fermentable plant fiber, respectively. Fecal microbiota composition was measured by 16S rRNA gene Illumina MiSeq sequencing (V3-V4 region) and Fluorescence in Situ Hybridization. Short chain fatty acids (SCFAs) and polyphenol microbial metabolites were determined. The three apple varieties significantly changed bacterial diversity, increased Actinobacteria relative abundance, acetate, propionate and total SCFAs (p < 0.05). Renetta Canada and Golden Delicious significantly decreased Bacteroidetes abundance and increased Proteobacteria proportion and bifidobacteria population (p < 0.05). Renetta Canada also increased Faecalibacterium prausnitzii, butyrate levels and polyphenol microbial metabolites (p < 0.05). Together, these data suggest that apples, particularly Renetta Canada, can induce substantial changes in microbiota composition and metabolic activity in vitro, which could be associated with potential benefits to human health. Human intervention studies are necessary to confirm these data and potential beneficial effects.
Project description:Optimisation of processing time and pre-treatments are crucial factors prior to apple drying to produce a high-quality product. The purpose of the present study was to test the utility of physical (hot-water, HWB and steam blanching, SB) and chemical (1% ascorbic acid, AA; and 1% citric acid, CA) treatments, alone or in combination in reducing surface discolouration as well as oxidative enzyme activity in apple slices (cv. Golden Delicious and Elstar) exposed to air at room temperature for 0, 30 and 60 min. The total colour change (?E) for Golden Delicious was equal to 2.38, 2.68, and 4.05 after 0, 30 and 60 min of air exposure, respectively. Dipping in AA solution (1% w/v) was found to be the best treatment to limit surface discolouration of both apple cultivars. The best heat treatments to inhibit polyphenol oxidase/peroxidase enzymes activity were 70 °C HWB for Golden Delicious and 60 °C HWB for Elstar slices, both in combination with a solution of 1% AA and 1% CA. The tested apple cultivars were found to require different treatments at minimum ambient air exposure to obtain the best surface colour condition.
Project description:Apples are a nutritious food source with significant amounts of polyphenols that contribute to human health and wellbeing, primarily as dietary antioxidants. Although numerous pre- and post-harvest factors can affect the composition of polyphenols in apples, genetics is presumed to play a major role because polyphenol concentration varies dramatically among apple cultivars. Here we investigated the genetic architecture of apple polyphenols by combining high performance liquid chromatography (HPLC) data with ~100,000 single nucleotide polymorphisms (SNPs) from two diverse apple populations. We found that polyphenols can vary in concentration by up to two orders of magnitude across cultivars, and that this dramatic variation was often predictable using genetic markers and frequently controlled by a small number of large effect genetic loci. Using GWAS, we identified candidate genes for the production of quercitrin, epicatechin, catechin, chlorogenic acid, 4-O-caffeoylquinic acid and procyanidins B1, B2, and C1. Our observation that a relatively simple genetic architecture underlies the dramatic variation of key polyphenols in apples suggests that breeders may be able to improve the nutritional value of apples through marker-assisted breeding or gene editing.
Project description:In terms of the quality of minimally processed fruit, flesh browning is fundamentally important in the development of an aesthetically unpleasant appearance, with consequent off-flavours. The development of browning depends on the enzymatic action of the polyphenol oxidase (PPO). In the 'Golden Delicious' apple genome ten PPO genes were initially identified and located on three main chromosomes (2, 5 and 10). Of these genes, one element in particular, here called Md-PPO, located on chromosome 10, was further investigated and genetically mapped in two apple progenies ('Fuji x Pink Lady' and 'Golden Delicious x Braeburn'). Both linkage maps, made up of 481 and 608 markers respectively, were then employed to find QTL regions associated with fruit flesh browning, allowing the detection of 25 QTLs related to several browning parameters. These were distributed over six linkage groups with LOD values spanning from 3.08 to 4.99 and showed a rate of phenotypic variance from 26.1 to 38.6%. Anchoring of these intervals to the apple genome led to the identification of several genes involved in polyphenol synthesis and cell wall metabolism. Finally, the expression profile of two specific candidate genes, up and downstream of the polyphenolic pathway, namely phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO), provided insight into flesh browning physiology. Md-PPO was further analyzed and two haplotypes were characterised and associated with fruit flesh browning in apple.
Project description:To learn how the endogenous polyphenols may play a role in fruit ripening and senescence, apple pulp discs were used as a model to study the influences of chlorogenic acid (CHA, a major polyphenol in apple pulp) on fruit ripening and senescence. Apple ('Golden Delicious') pulp discs prepared from pre-climacteric fruit were treated with 50 mg L(-1) CHA and incubated in flasks with 10 mM MES buffer (pH 6.0, 11% sorbitol). Compared to the control samples, treatment with CHA significantly reduced ethylene production and respiration rate, and enhanced levels of firmness and soluble solids content of the pulp discs during incubation at 25°C. These results suggested that CHA could retard senescence of the apple pulp discs. Proteomics analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry (MALDI-TOF/TOF) revealed that the expressions of several key proteins correlated to fruit ripening and senescence were affected by the treatment with CHA. Further study showed that treating the pulp discs with CHA remarkably reduced levels of lipoxygenase, ?-galactosidase, NADP-malic enzyme, and enzymatic activities of lipoxygenase and UDP-glucose pyrophosphorylase, all of which are known as promoters of fruit ripening and senescence. These results could provide new insights into the functions of endogenous phenolic compounds in fruit ripening and senescence.
Project description:Apple polyphenols have been studied for various beneficial bioactivities. Especially interesting are traditional, old varieties of apples for which some initial studies have suggested significant bioactivities, but they are still not completely understood. Polyphenol bioactivities can be affected by interactions with dietary fibers such as ?-glucans. The aim of this study was to investigate for the first time interactions between individual polyphenols from traditional, old apple varieties ("Boži?nica" and "Batulenka") and ?-glucans by studying the adsorption process. Polyphenols were extracted from the peel and flesh of traditional apples by using an ultrasonic bath and characterized with high-performance liquid chromatography. The amounts of adsorbed (qe) and un-adsorbed (ce) polyphenols were modeled with adsorption isotherms (Langmuir, Dubinin-Radushkevich, and Hill) by using improved non-linear fitting in a novel R algorithm, developed specifically for the modeling of adsorption isotherms. Polyphenols adsorbed onto ?-glucan from 9 to 203 (peel, "Boži?nica"), 1 to 484 (peel, "Batulenka"), 5 to 160 (flesh, "Boži?nica"), and 19 to 28 mg g-1 (flesh, "Batulenka"). The adsorption was concentration dependent (polyphenols present in higher amount adsorbed in higher amounts). Physical sorption can be suggested. Polyphenols from traditional apples adsorb onto ?-glucan and should be further studied.
Project description:Triterpene acid and phenolic constituents from nine ancient varieties of apple (Malus domestica) fruits cultivated in Fanna, Friuli Venezia Giulia region, northeast Italy, were analyzed and compared with four commercial apples ('Golden Delicious', 'Red Delicious', 'Granny Smith' and 'Royal Gala'). Total phenolic and flavonoid contents were measured by spectrophotometric assays. The quali-quantitative fingerprint of secondary metabolites including triterpene acid was obtained by LC-DAD-(ESI)-MS and LC-(APCI)-MS, respectively. Based on the two LC-MS datasets, multivariate analysis was used to compare the composition of ancient fruit varieties with those of four commercial apples. Significant differences related mainly to the pattern of triterpene acids were found. Pomolic, euscaphyc, maslinic and ursolic acids are the most abundant triterpene in ancient varieties pulps and peels, while ursolic and oleanolic acids were prevalent in the commercial fruits. Also, the content of the phenolic compounds phloretin-2-O-xyloglucoside and quercetin-3-O-arabinoside was greater in ancient apple varieties. The antioxidant (radical scavenging, reducing power, metal chelating and phosphomolybdenum assays) and enzyme inhibitory effects (against cholinesterase, tyrosinase, amylase and glucosidase) of the samples were investigated in vitro. Antioxidant assays showed that the peels were more active than pulps. However, all the samples exhibited similar enzyme inhibitory effects. Ancient Friuli Venezia Giulia apple cultivars can be a source of chlorogenic acid and various triterpene acids, which are known for their potential anti-inflammatory activity and beneficial effects on lipid and glucose metabolism. Our results make these ancient varieties suitable for the development of new nutraceutical ingredients.
Project description:Background:Fruit pomaces are by-products rich in polyphenol compounds and dietary fiber. They seem to play an important role in regulating the gut microbiota, morphology and physiology. The aim of this study was to assess whether apple (A), blackurrant (B) or strawberry (S) pomaces could be suitable ingredients in broiler diets and their effect on gut health. A total of 480 male broilers were randomly allotted to 8 dietary treatments with lower (3%-L) or higher (6%-H) dietary fiber content: two control groups (CL/CH), two A diets (AL/AH), two B diets (BL/BH), two S diets (SL/SH). Diet and fruit pomaces were chemically analyzed to assess polyphenol concentration and fibre fraction content. After the evaluation of growth performance, 6 birds/group were slaughtered at 35?days of age. Morphometric and histopathological investigations were performed on duodenum, jejunum and ileum. Excreta were collected to perform microbiota evaluation by 16S DNA sequencing. Weight, viscosity, enzymatic activity, short chain fatty acid (SCFAs) and ammonia concentration were determined in ileum and/or ceca content. Results:A pomace and A diets showed the lowest polyphenol content and the highest content of soluble fibre fraction. No significant differences were observed for growth performance, gut morphometry and histopathology (P?>?0.05). Dietary fruit pomace inclusion increased the weight of ileum and ceca and the ileum digesta viscosity (P?<?0.05). In the ileum, A and S groups showed lower bacterial ?-glucosidase activity than C groups. Moreover, small intestine SCFAs concentration was higher in fruit pomaces diets (P?<?0.05). In ceca, B and S groups showed lower ammonia concentration and higher SCFAs than C. Dietary treatments also influenced the activity of ?-glucosidase, ?-galactosidase, ?-galactosidase ?-glucuronidase and xylase. Regarding microbiota, at phylum level, Firmicutes were differentially abundant across treatment (maximum for C and minimum in S, FDR?>?0.05). At genus level, an increase of Weissella in AH and Erwinia in S/B diets, as well as a decrease of Lactobacillus in all fruit pomace groups were recorded (P?<?0.05). Conclusions:Fruit pomaces could be suitable ingredients in poultry nutrition even if further studies are needed to better understand which doses is more recommended to avoid negative effects on gut microbiota.
Project description:This study examined the effect of debagging time on color and flavor / taste compounds in the non-red apple cultivar 'Golden Delicious' and red cultivar 'Qinguan' at mid and late stages of fruit development. Debagging briefly improved the red color in both cultivars, the peel of 'Golden Delicious' presenting pale-pink hue. However, rapid anthocyanin accumulation occurred in apple peel at a specific time (after 179 days after flowering (DAF) in 'Qinguan') and was unaltered by debagging time in the red cultivar 'Qinguan'. Furthermore, untimely debagging had a detrimental effect on the content of anthocyanin. All sugars increased and organic acids decreased in apple pulp at mid to late stages of development. Bagging treatment reduced the content of most sugars and organic acids, as well as, the overall total. However, glucose and citric acid contents were higher in bagged fruit than non-bagged fruit; the maximum occurred in T7 treatment that was no-debagging at DAF 159 / 196 ('Golden delicious' / 'Qinguan'), i.e., 24.35 and 0.07 mg g-1 FW in 'Golden delicious', and 38.86 and 0.06 mg g-1 FW in 'Qinguan', respectively. In a word, bagging treatment can alter the pattern of peel color development in apple fruit; however, it remains difficult to alter the timing of rapid anthocyanin accumulation as it is regulated solely by development. Moreover, bagging treatment reduced the total accumulation of sugars and organic acids, and even the over total in pulp, but increased the glucose and citric acid contents in apple pulp.
Project description:The apple (Malus × domestica Borkh.) cultivar 'Su Shuai' exhibits greater disease resistance, shorter internodes and lighter fruit flavor compared with its parents 'Golden Delicious' and 'Indo'. To obtain a comprehensive overview of the sequence variation in these three horticultural traits, the genomes of 'Su Shuai' and 'Indo' were resequenced using next-generation sequencing and compared to the genome of 'Golden Delicious'. A wide range of genetic variations were detected, including 2?454?406 and 18?749?349 single nucleotide polymorphism (SNP) and 59?547 and 50?143 structural variants (SVs) in the 'Indo' and 'Su Shuai' genomes, respectively. Among the SVs in 'Su Shuai', 17 genes related to disease resistance, 10 genes related to Gibberellin (GA) and 19 genes associated with fruit flavor were identified. The expression patterns of eight of the SV genes were examined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results of this study illustrate the genomic variation in these cultivars and provide evidence for a genetic basis for the horticultural traits of disease resistance, short internodes and lighter flavor exhibited in these cultivars. These results provide a genetic basis for the phenotypic characteristics of 'Su Shuai' and, as such, these SVs could serve as gene-specific molecular markers in maker-assisted breeding of apples.
Project description:BACKGROUND:Apples are rich in bioactive polyphenols and fiber. Evidence suggests that consumption of apples or their bioactive components is associated with beneficial effects on lipid metabolism and other markers of cardiovascular disease (CVD). However, adequately powered randomized controlled trials are necessary to confirm these data and explore the mechanisms. OBJECTIVE:We aimed to determine the effects of apple consumption on circulating lipids, vascular function, and other CVD risk markers. METHODS:The trial was a randomized, controlled, crossover, intervention study. Healthy mildly hypercholesterolemic volunteers (23 women, 17 men), with a mean ± SD BMI 25.3 ± 3.7 kg/m2 and age 51 ± 11 y, consumed 2 apples/d [Renetta Canada, rich in proanthocyanidins (PAs)] or a sugar- and energy-matched apple control beverage (CB) for 8 wk each, separated by a 4-wk washout period. Fasted blood was collected before and after each treatment. Serum lipids, glucose, insulin, bile acids, and endothelial and inflammation biomarkers were measured, in addition to microvascular reactivity, using laser Doppler imaging with iontophoresis, and arterial stiffness, using pulse wave analysis. RESULTS:Whole apple (WA) consumption decreased serum total (WA: 5.89 mmol/L; CB: 6.11 mmol/L; P = 0.006) and LDL cholesterol (WA: 3.72 mmol/L; CB: 3.86 mmol/L; P = 0.031), triacylglycerol (WA: 1.17 mmol/L; CB: 1.30 mmol/L; P = 0.021), and intercellular cell adhesion molecule-1 (WA: 153.9 ng/mL; CB: 159.4 ng/mL; P = 0.028), and increased serum uric acid (WA: 341.4 ?mol/L; CB: 330 ?mol/L; P = 0.020) compared with the CB. The response to endothelium-dependent microvascular vasodilation was greater after the apples [WA: 853 perfusion units (PU), CB: 760 PU; P = 0.037] than after the CB. Apples had no effect on blood pressure or other CVD markers. CONCLUSIONS:These data support beneficial hypocholesterolemic and vascular effects of the daily consumption of PA-rich apples by mildly hypercholesterolemic individuals.This trial was registered at clinicaltrials.gov as NCT01988389.