The Action Mechanisms, Anti-Cancer and Antibiotic-Modulation Potential of Vaccinium myrtillus L. Extract.

BACKGROUND: Herbal medicinal products containing Vaccinium myrtillus L. (bilberry) fruits and fruit extracts are widely available in the market. Although bilberry leaves and stems are considered as bio-waste, they contain much higher levels of phenolic compounds than fruits. The study aimed to investigate the antimicrobial and anticancer potential of aerial part extracts from Vaccinium myrtillus L. (V. myrtillus, VM) plants harvested at high altitudes in Armenian landscape and characterize the bioactive phytochemicals. MATERIAL AND METHODS: For evaluation of antioxidant properties, chemical-based tests (total phenolic and flavonoid content, and antiradical activity in 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) tests) and cellular antioxidant activity (CAA) assay were applied. Genotoxicity and anticancer properties of the extract alone and in combination with fluorouracil were explored in human cancer and normal cell lines. Antibacterial properties of V. myrtillus extract alone and in combination with antibiotics, as well as their effect on proton-flux rate through cell membrane were explored on bacterial strains. The characterization of active phytochemicals was done using Liquid Chromatography-Quadrupole-Orbitrap High-Resolution Mass Spectrometry (LC-Q-Orbitrap HRMS). RESULTS: The V. myrtillus aerial part extract demonstrated promising antioxidant properties in all tests. The selective cytotoxic activity was documented against various cancer cell lines (human colon adenocarcinoma (HT29), human breast cancer (MCF-7) and human cervical carcinoma (HeLa)), while it did not inhibit the growth of tested human normal primary renal mixed epithelial cells (HREC) even at 10-fold higher concentrations. The extract did not have genotoxic properties in comet assay making it a potential source for the development of anticancer preparations. The investigated extract did not directly inhibit the growth of Escherichia coli (E. coli) and Salmonella typhimurium (S. typhimurium) strains at up to 1 mg/mL concentration. However, V. myrtillus extract enhanced the kanamycin intake and increased its efficiency against E. coli strain. The phytochemical characterization of the extract showed the presence of different groups of phenolics. CONCLUSIONS: Based on obtained data, we suggest the aerial parts of the V. myrtillus plant as an alternative source of bioactive natural products for food supplements, nutraceuticals, functional foods and medicine.

Antidepressant effects of p-coumaric acid isolated from Vaccinium bracteatum leaves extract on chronic restraint stress mouse model and antagonism of serotonin 6 receptor in vitro.

BACKGROUND: Vaccinium bracteatum Thunb. leaves (VBL) are used in traditional herbal medicines to treat various biological diseases. p-coumaric acid (CA), the main active component of VBL, has neuroprotective effects against corticosterone-induced damage in vitro. However, the effects of CA on immobility induced by chronic restraint stress (CRS) in a mouse model and 5-HT receptor activity have not been investigated. HYPOTHESIS/PURPOSE: We investigated the antagonistic effects of VBL, NET-D1602, and the three components of Gαs protein-coupled 5-HT receptors. Additionally, we identified the effects and mechanism of action of CA, the active component of NET-D1602, in the CRS-exposed model. METHODS: For in vitro analyses, we used 1321N1 cells stably expressing human 5-HT6 receptors and CHO-K1 expressing human 5-HT4 or 5-HT7 receptors cell lines to study the mechanism of action. For in vivo analyses, CRS-exposed mice were orally administered CA (10, 50, or 100 mg/kg) daily for 21 consecutive days. The effects of CA were analyzed by assessing behavioral changes using a forced swim test (FST), measuring levels of hypothalamic-pituitary-adrenal (HPA) axis-related hormones in ntial therapeutic effects as 5-HT6 receptor antagonists for neurodegenerative diseases and depressioserum, and acetylcholinesterase (AChE), monoamines, including 5-HT, dopamine, and norepinephrine, using enzyme-linked immunosorbent assay kits. The underlying molecular mechanisms of the serotonin transporter (SERT), monoamine oxidase A (MAO-A), and extracellular signal-regulated kinase (ERK)/protein kinase B (Akt)/mTORC1 signaling were detected using western blotting. RESULTS: CA was confirmed to be an active component in the antagonistic effects of NET-D1602 on 5-HT6 receptor activity through decreases in cAMP and ERK1/2 phosphorylation. Moreover, CRS-exposed mice treated with CA showed a significantly reduced immobility time in the FST. CA also significantly decreased corticosterone, corticotropin-releasing hormone (CRH), and adrenocorticotropic hormone (ACTH) levels. CA enhanced 5-HT, dopamine, and norepinephrine levels in the hippocampus (HC) and prefrontal cortex (PFC) but decreased MAO-A and SERT protein levels. Similarly, CA significantly upregulated the ERK, Ca2+/calmodulin-dependent protein kinase II (CaMKII), Akt/mTOR/p70S6K/S6 signaling pathways in both HC and the PFC. CONCLUSION: CA contained in NET-D1602 may play the antidepressant effects against CRS-induced depression-like mechanism and the selective antagonist effect of 5-HT6 receptor.

Phytonutrients of bilberry fruit and saskatoon berry in the prevention and treatment of dyslipidemia.

The epidemiologic studies from the recent years indicate that high consumption of foods rich in bioactive compounds has a positive effect on human health and could diminish the risk of numerous diseases, such as cancer, heart disease, stroke, Alzheimer's disease, diabetes, cataracts, and even diseases related to age. From all species of fruit, definitely consumption of berries due to its high content of bioactive constituents prevents the risk of cardiovascular disease, oxidative stress and diabetes. The primary phenolic compounds in berries are flavonoids, particularly the anthocyanins. They have potential preventative and therapeutic effects on many diseases such as cancers, inflammation and cardiovascular diseases, obesity, neurodegenerative pathologies, and muscular degeneration. Bilberry fruits have been an important part of local diets in many countries, including Slovakia. They are valued for their pleasant taste and aroma and are often processed into jams, preserves, juices, and alcoholic beverages. In the last two decades, the Saskatoon berry has been cultivated in many parts of the world for its suitability for various food products and due to its high content of nutrients and polyphenols. Cardiovascular disease (CVD) remains the world's leading cause of morbidity and mortality. Dyslipidemia, which results from one or more abnormalities of blood lipids metabolism, remains a major key factor for progression of CVD and leads to the development of atherosclerotic plaques. The aim of this review is to compare and summarize the research evidence on the potential of bilberries and saskatoon berries with an emphasis on recent studies in humans in improving cardiovascular risk factors especially dyslipidemia.

Insights into sugar metabolism during bilberry (Vaccinium myrtillus L.) fruit development.

Bilberry fruit is regarded as one of the best natural sources of anthocyanins and is widely explored for its health-beneficial compounds. Besides anthocyanins, one of the major attributes that determine the berry quality is the accumulation of sugars that provide sweetness and flavor to ripening fruit. In this study, we have identified 25 sugar metabolism-related genes in bilberry, including invertases (INVs), hexokinases (HKs), fructokinases (FKs), sucrose synthases (SSs), sucrose phosphate synthases (SPSs), and sucrose phosphate phosphatases (SPPs). The results indicate that isoforms of the identified genes are expressed differentially during berry development, suggesting specialized functions. The highest sugar content was found in ripe berries, with fructose and glucose dominating accompanied by low sucrose amount. The related enzyme activities during berry development and ripening were further analyzed to understand the molecular mechanism of sugar accumulation. The activity of INVs in the cell wall and vacuole increased toward ripe berries. Amylase activity involved in starch metabolism was not detected in unripe berries but was found in ripe berries. Sucrose resynthesizing SS enzyme activity was detected upon early ripening and had the highest activity in ripe berries. Interestingly, our transcriptome data showed that supplemental irradiation with red and blue light triggered upregulation of several sugar metabolism-related genes, including α- and ß-amylases. Also, differential expression patterns in responses to red and blue light were found across sucrose, galactose, and sugar-alcohol metabolism. Our enzymological and transcriptional data provide new understanding of the bilberry fruit sugar metabolism having major effect on fruit quality.

Assessing compatibility of excipients selected for a sustained release formulation of bilberry leaf extract

Abstract The study is aimed to assess the compatibility of bilberry leaf powder extract (BLPE) with six excipients selected for sustained-release (SR) tablet formulation. The BLPE was obtained with the addition of L-arginine and Myo-inositol as the carriers. Thermogravimetric (TG-DTG) analysis and Fourier-transform infrared spectroscopy (FTIR), supported by Pearson correlation analysis, were applied to detect possible interactions in the binary mixtures (1:1) of the BLPE with each excipient. The TG-DTG showed some deviations in the thermal behavior of the BLPE / excipient mixtures. However, only the thermal behavior of magnesium stearate in the mixture significantly differed from individual samples, which suggested chemical interaction for this excipient. The FTIR analysis confirmed that the BLPE is compatible with Eudragit L100, Methocel K4M, Methocel K100LV, Avicel PH-101, and Plasdone S-630. Whereas it undergoes solid-state chemical interaction in the binary mixture with magnesium stearate. According to the FTIR-spectra, it is suggested that this interaction results in the formation of stearic acid and alkalization of the medium. These findings evidence for the possibility of using TG-DTG analysis as an independent thermal technique for compatibility studies and also confirm the earlier reported interaction of basic lubricants, e.g., stearic salts, with active ingredients containing amino groups.

Insulin receptors in the CA1 field of hippocampus and selected blood parameters in diabetic rats fed with bilberry fruit.

INTRODUCTION: Bilberry fruit is believed to be a promising factor in the treatment of diabetes mellitus. Chronic hyperglycaemia affects the function of the central nervous system, which may be manifested as changes in hypothalamic insulin signalling. MATERIAL AND METHODS: Using DPPH and ABTS assays, total phenolic content in bilberry fruit and its antioxidant activities were examined. The selected biochemical parameters of blood (glucose, fructosamine, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides), as well as the expression of insulin receptors, were studied in the hippocampal CA1 field of healthy and diabetic (streptozotocin-induced; 60 mg kg-1 body weight) Wistar rats fed with bilberry fruit (16 g kg-1 body weight per day; 6 weeks), as well as of the corresponding control groups. RESULTS: Biochemical analyses revealed ambiguous results, but a significantly (P<0.05) decrease in the level of LDL-cholesterol was observed in the group of healthy rats supplemented with bilberry pulp after 6 weeks of the treatment. There was also a difference (P<0.05) in the level of LDL-cholesterol in the mentioned healthy animals fed with bilberry, versus the healthy control group. An increased number of insulin receptors-immunoreactive neurons as well as nerve fibres in the CA1 field of diabetic rats fed with bilberry fruit was also found. CONCLUSIONS: An inclusion of bilberry fruit in the daily diet during the course of diabetes can lead to plasticity of hippocampal neurons/nerve fibres, manifested by changes in insulin receptors expression. Whether or not the observed changes had protective effects (by reducing damages caused by diabetes mellitus) on the function of the central nervous system neurons needs further study.

Analysis of composition, morphology, and biosynthesis of cuticular wax in wild type bilberry (Vaccinium myrtillus L.) and its glossy mutant.

In this study, cuticular wax load, its chemical composition, and biosynthesis, was studied during development of wild type (WT) bilberry fruit and its natural glossy type (GT) mutant. GT fruit cuticular wax load was comparable with WT fruits. In both, the proportion of triterpenoids decreased during fruit development concomitant with increasing proportions of total aliphatic compounds. In GT fruit, a higher proportion of triterpenoids in cuticular wax was accompanied by a lower proportion of fatty acids and ketones compared to WT fruit as well as lower density of crystalloid structures on berry surfaces. Our results suggest that the glossy phenotype could be caused by the absence of rod-like structures in GT fruit associated with reduction in proportions of ketones and fatty acids in the cuticular wax. Especially CER26-like, FAR2, CER3-like, LTP, MIXTA, and BAS genes showed fruit skin preferential expression patterns indicating their role in cuticular wax biosynthesis and secretion.

Plant growth shapes the effects of elevation on the content and variability of flavonoids in subalpine bilberry stands.

The study of morphological and physiological responses of shrubs to climate is crucial for the understanding of future scenarios regarding climate change. In this light, studying shrub growth and physiological acclimation along an elevation gradient might be insightful. The phenolic metabolic pathway represents a powerful tool to interpret such processes. In the South-Eastern Alps, we investigated the relationships between elevation, plant traits (i.e. age, xylem ring width, annual shoot length), plant-plant interaction (i.e. shrub cover) and flavonoids in Vaccinium myrtillus L. (leaves, berries) in stands above the treeline. The relationships were parsed within causal networks using a confirmatory path analysis. Elevation was the main driver of V. myrtillus growth, having both direct and indirect effects on the leaf flavonoid content, but this was less evident for berries. In particular, the content of foliar flavonoids showed a peak at mid-elevation and where the growth of xylem rings was intermediate, while it decreased in stands with higher shoot length. Flavonoid content variability of both leaves and berries was affected by elevation and shoot length. In berries, flavonoid variability was further related to all growth traits and shrub cover. These findings evidence that flavonoid content is influenced by both elevation and growth traits of V. myrtillus, often showing non-linear relationships. These results suggest a trait-mediated response of this plant to climate conditions as a result of trade-offs between plant growth, plant defence, environmental stress and nutrient/resource availability.

Development of new bilberry (Vaccinium myrtillus L.) based snacks: Nutritional, chemical and bioactive features.

The sustainable exploitation of fruit and cereal processing is being conducted to produce novel food products with promising nutritional properties and high content in added value compounds. Herein, three bilberry fruit-based snacks supplemented with edible petals and fruits were characterized for their nutritional properties and chemical composition. The phenolic profile, antioxidant, antibacterial and hepatotoxic properties were analyzed. Protein (3-4 g/100 g dw) and carbohydrates (94.3-94.8 g/100 g dw) represented the major macronutrients. The combination of bilberry fruits with edible petals, calendula and rose, improved the nutritional and phytochemical input in organic acids and tocopherols content, respectively. Also, the supplementation with apple and goji fruits provided higher content in phenolic acids and anthocyanins (up to a 9-fold higher concentration, 199.7 µg/g of extract), also resulting in a higher antioxidant and antibacterial activities. The results obtained can contribute for the development of novel sustainable and healthier snacks for the food industry.

Effects of Anthocyanin Extracts from Bilberry (Vaccinium myrtillus L.) and Purple Potato (Solanum tuberosum L. var. 'Synkeä Sakari') on the Plasma Metabolomic Profile of Zucker Diabetic Fatty Rats.

This study compared the effects of the nonacylated and acylated anthocyanin-rich extracts on plasma metabolic profiles of Zucker diabetic fatty rats. The rats were fed with the nonacylated anthocyanin extract from bilberries (NAAB) or the acylated anthocyanin extract from purple potatoes (AAPP) at daily doses of 25 and 50 mg/kg body weight for 8 weeks. 1H NMR metabolomics was used to study the changes in plasma metabolites. A reduced fasting plasma glucose level was seen in all anthocyanin-fed groups, especially in the groups fed with NAAB. Both NAAB and AAPP decreased the levels of branched-chain amino acids and improved lipid profiles. AAPP increased the glutamine/glutamate ratio and decreased the levels of glycerol and metabolites involved in glycolysis, suggesting improved insulin sensitivity, gluconeogenesis, and glycolysis. AAPP decreased the hepatic TBC1D1 and G6PC messenger RNA level, suggesting regulation of gluconeogenesis and lipogenesis. This study indicated that AAPP and NAAB affected the plasma metabolic profile of diabetic rats differently.

Changes in the Proanthocyanidin Composition and Related Gene Expression in Bilberry (Vaccinium myrtillus L.) Tissues.

Berries of genus Vaccinium are rich in flavonoids and proanthocyanidins (PAs). We studied the PA composition and biosynthesis in bilberry (Vaccinium myrtillus L.) tissues and during fruit development. Soluble PAs, analyzed by UHPLC-MS/MS, were most abundant in stem and rhizome with the mean PA polymerization level varying between 4 and 6 in all tissues. Both A- and B-type PAs were present in all tissues. Procyanidin subunits were more common than prodelphinidin subunits in PAs. During fruit ripening, the amount of procyanidin subunits decreased while prodelphinidin subunits and F3'5'H expression increased, indicating a shift in biosynthesis toward the delphinidin branch of the flavonoid pathway. Epicatechin was the most abundant flavan-3-ol in all tissues. Expression of ANR and three isolated LAR genes, analyzed by qRT-PCR, showed connection to accumulation of PAs and flavan-3-ols biosynthesized from different flavonoid branches. Insoluble PAs accumulated during berry development, suggesting that PAs are not recycled after biosynthesis.

Pyranoanthocyanins in bilberry (Vaccinium myrtillus L.) wines fermented with Schizosaccharomyces pombe and their evolution during aging.

Fifteen vitisin A-type pyranoanthocyanins (vAPs) were determined in bilberry wines fermented with Saccharomyces cerevisiae and Schizosaccharomyces pombe by HPLC-DAD and UPLC-DAD-ESI-MS/MS. The fermentation involving S. pombe enhanced the production of vAPs compared to the fermentation with pure S. cerevisiae. The formation of vAPs correlated significantly with the decrease in the content of monomeric anthocyanins and pyruvic acid during 12 months of aging. vAPs were more stable than their corresponding monomeric anthocyanins. Methylation in the B-ring and glycosylation with galactose and arabinose further improved the stability of vAPs. Aging for 12 months led to depletion of pyruvic acid and reduction of over 50% of monomeric anthocyanins. The content of vAPs increased by 26-54% during the first six months of aging, followed by a 2.2-10.2% reduction over the following six months. More residual pyruvic acid in S. pombe wines after fermentation consequently enhanced the generation of vAPs during aging.

Comparative investigation on metabolite changes in 'wu mi' production by Vaccinium bracteatum Thunb. leaves based on multivariate data analysis using UPLC-QToF-MS.

Vaccinium bracteatum Thunb. leaves (VBTL) are used to juicing and dye rice to produce 'Wu mi', which displays a deep blue color. However, little information is known about the formation mechanism of the pigments. In this research, non-targeted metabolic profiling of VBTL and VBTL juice samples at different growth stages was studied for searching the pigment precursors through UPLC-QToF-MS and multivariate data analysis. The results showed the L* and b* values of 'Wu mi' produced by spring leaves (stages of 2WAB, 4WAB and 6WAB) were significantly lower than other growth stages. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) of the VBTL and VBTL juice samples showed distinct classifications. Further variable importance in projection (VIP) plot in PLS-DA model demonstrated the discriminatory potential biomarkers between VBTL and VBTL juice samples. Some of the identified biomarkers were tentatively identified as the precursor compounds of the iridoid-derived pigments.

Chlorogenic and caffeic acids in 64 fruits consumed in Brazil.

This study presents unpublished data on the chlorogenic acid and caffeic acid content in fruits. Sixty-four fruits consumed in Brazil, most of which were produced domestically, were evaluated based on their levels of 3-caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and caffeic acid. The study investigated 15 fruits that have not been reported in the literature in relation to these compounds, including several native species. The highest concentration of mono-caffeoylquinic acid was observed in strawberry, cherry, bilberry, quince and mulberry, while the dicaffeoylquinic acid was present with highest concentration in kumquat, passion fruit and sweet granadilla. Regarding caffeic acid, the highest content was found in bilberry and yellow pitaya. Considering the sum of the concentrations of these compounds, quince, cherry, bilberry, mulberry and sweet granadilla were exceptional, with concentrations between 200.0 and 569.7 mg kg-1.

Transcriptional profiling of methyl jasmonate-induced defense responses in bilberry (Vaccinium myrtillus L.).

BACKGROUND: Bilberry (Vaccinium myrtillus L.) is one of the most abundant wild berries in the Northern European ecosystems. This species plays an important ecological role as a food source for many vertebrate and invertebrate herbivores. It is also well-recognized for its bioactive compounds, particularly substances involved in natural defenses against herbivory. These defenses are known to be initiated by leaf damage (e.g. chewing by insects) and mediated by activation of the jasmonic acid (JA) signaling pathway. This pathway can be activated by exogenous application of methyl jasmonate (MeJA), the volatile derivative of JA, which is often used to stimulate plant defense responses in studies of plant-herbivore interactions at ecological, biochemical, and molecular organismal levels. As a proxy for herbivore damage, wild V. myrtillus plants were treated in the field with MeJA and changes in gene expression were compared to untreated plants. RESULTS: The de novo transcriptome assembly consisted of 231,887 unigenes. Nearly 71% of the unigenes were annotated in at least one of the databases interrogated. Differentially expressed genes (DEGs), between MeJA-treated and untreated control bilberry plants were identified using DESeq. A total of 3590 DEGs were identified between the treated and control plants, with 2013 DEGs upregulated and 1577 downregulated. The majority of the DEGs identified were associated with primary and secondary metabolism pathways in plants. DEGs associated with growth (e.g. those encoding photosynthesis-related components) and reproduction (e.g. flowering control genes) were frequently down-regulated while those associated with defense (e.g. encoding enzymes involved in biosynthesis of flavonoids, lignin compounds, and deterrent/repellent volatile organic compounds) were up-regulated in the MeJA treated plants. CONCLUSIONS: Ecological studies are often limited by controlled conditions to reduce the impact of environmental effects. The results from this study support the hypothesis that bilberry plants, growing in natural conditions, shift resources from growth and reproduction to defenses while in a MeJA-induced state, as when under insect attack. This study highlights the occurrence of this trade-off at the transcriptional level in a realistic field scenario and supports published field observations wherein plant growth is retarded and defenses are upregulated.

Volatile composition of bilberry wines fermented with non-Saccharomyces and Saccharomyces yeasts in pure, sequential and simultaneous inoculations.

Bilberry (Vaccinium myrtillus L.) juice was fermented with Torulaspora delbrueckii (TD291 and TD70526) and Schizosaccharomyces pombe (SP3796 and SP70572) in pure fermentation as well as in sequential and simultaneous inoculations with Saccharomyces cerevisiae 1116 (SC1116). Altogether, 56 volatile compounds were identified and semi-quantified with HS-SPME-GC/MS in bilberry products. Yeast fermentation prominently enhanced the aroma complexity of bilberry with a sharp increase in alcohols, esters, aldehydes, and acetals. Compared to S. cerevisiae, T. delbrueckii produced less ethanol but more fusel alcohols that potentially enhance "alcohol" and "nail polish" odors in TD70526 and less "fruity" esters in TD291. SP70572 resulted in high productions of undesirable compounds of acetoin and acetaldehyde but a low content of higher alcohols and esters, SP3796 produced a high content of fatty acid ethyl esters and acetoin. In comparison with monoculture of non-Saccharomyces yeast, sequential and simultaneous cultures of S. pombe and S. cerevisiae significantly decreased the content of acetoin while increased the relative level of esters; sequential cultures of T. delbrueckii and S. cerevisiae remarkably increased the concentration of acetaldehyde; simultaneous inoculations of S. cerevisiae with TD70526 and TD291 significantly decreased the content of fusel alcohols and increased the content of esters, respectively. The findings suggested that non-Saccharomyces yeasts possess the potential to affect and modulate the aromatic profile of fermented bilberry products. Sequential and simultaneous inoculations with S. pombe strains and S. cerevisiae as well as simultaneous fermentation using T. delbrueckii strains and S. cerevisiae are optimal strategies to positively influence the aroma profile of bilberry wines.

Effect of anthocyanins on lipid oxidation and microbial spoilage in value-added emulsions with bilberry seed oil, anthocyanins and cold set whey protein hydrogels.

The objective of this work was to explore the storage properties of a structured oil-in-water emulsion containing both water- and fat-soluble bioactive compounds from bilberries (Vaccinium myrtillus L.). Bilberry seed oil (BSO) was dispersed in a continuous aqueous phase of anthocyanins (AC) and whey protein isolate. The microstructure was evaluated using light microscopy and the effect of anthocyanins on lipid oxidation and microbial growth was investigated. The results showed that it was possible to generate a stable emulsion structure that resisted phase separation during 25 weeks of storage. Gas chromatography-mass spectrometry measurements of the fatty acids in the BSO during storage showed that AC had a protective effect against lipid oxidation. The AC did not have an antimicrobial effect against the investigated strains Zygosaccharomyces bailii (ATCC 42476) and Aspergillus niger (ATCC 6275 (M68)).

Chemical composition of bilberry wine fermented with non-Saccharomyces yeasts (Torulaspora delbrueckii and Schizosaccharomyces pombe) and Saccharomyces cerevisiae in pure, sequential and mixed fermentations.

This study evaluated the effects of fermentation with pure cultures of Torulaspora delbrueckii (TD291 and TD70526) and Schizosaccharomyces pombe (SP3796 and SP70572), as well as in sequential and mixed inoculations with Saccharomyces cerevisiae, on the chemical composition of bilberry wine. In comparison to the bilberry wines produced by pure and sequential fermentations, mixed cultures produced bilberry wines with more ethanol, higher pH values, higher percentages of red and yellow shade, but less glycerol and acetaldehyde. Higher values of color intensity and bluish parameter were found in products of pure fermentations with non-Saccharomyces yeasts. Compared to S. cerevisiae, T. delbrueckii contributed to the reduction of ethanol and acetic acid while increasing the content of succinic acid, lactic acid and higher alcohols; S. pombe consumed malic acid almost completely and produced more glycerol, acetaldehyde and/or pyruvic acid. Fermentation with SP70572 had the highest amounts of anthocyanins and hydroxycinnamic acids derivatives.

Daily Consumption of Bilberry ( Vaccinium myrtillus L.) Extracts Increases the Absorption Rate of Anthocyanins in Rats.

The effects of daily consumption of anthocyanins on bioavailability has remained unclear. In this study, we evaluated whether daily consumption affects the absorption rate of anthocyanins in rats when consumed during the active and sleep phase. Eighty rats were randomly divided into two groups. The first group consumed AIN-93G control diets, and the second group consumed AIN-93G diets containing 1% bilberry extract for 2 weeks. After 12 h fast, anthocyanins were not detected in plasma of rats. Bilberry extract (500 mg/kg body weight) was then orally administered at the beginning of the diurnal light period (ZT0, sleep phase) or at the end of the diurnal light period (ZT12, active phase). Blood concentrations of anthocyanins peaked 1 h after administration in both groups. Maximum blood concentration in rats that consumed bilberry extract daily (852 nM) was higher than that in control rats (630 nM) when the extract was administered at ZT0 but not at ZT12. Daily consumption of anthocyanins increases their absorption rate, but this effect is limited to the beginning of the sleep phase.

Recognition of candidate transcription factors related to bilberry fruit ripening by de novo transcriptome and qRT-PCR analyses.

Bilberry (Vaccinium myrtillus L.) fruits are an excellent natural resource for human diet because of their special flavor, taste and nutritional value as well as medical properties. Bilberries are recognized for their high anthocyanin content and many of the genes involved in the anthocyanin biosynthesis have been characterized. So far, neither genomic nor RNA-seq data have been available for the species. In the present study, we de novo sequenced two bilberry fruit developmental stages, unripe green (G) and ripening (R). A total of 57,919 unigenes were assembled of which 80.2% were annotated against six public protein databases. The transcriptome served as exploratory data to identify putative transcription factors related to fruit ripening. Differentially expressed genes (DEGs) between G and R stages were prominently upregulated in R stage with the functional annotation indicating their main roles in active metabolism and catalysis. The unigenes encoding putative ripening-related regulatory genes, including members of NAC, WRKY, LOB, ERF, ARF and ABI families, were analysed by qRT-PCR at five bilberry developmental stages. Our de novo transcriptome database contributes to the understanding of the regulatory network associated with the fruit ripening in bilberry and provides the first dataset for wild Vaccinium species acquired by NGS technology.