Strategy of Coniferous Needle Biorefinery into Value-Added Products to Implement Circular Bioeconomy Concepts in Forestry Side Stream Utilization.

Sustainable development goals require a reduction in the existing heavy reliance on fossil resources. Forestry can be considered a key resource for the bioeconomy, providing timber, energy, chemicals (including fine chemicals), and various other products. Besides the main product, timber, forestry generates significant amounts of different biomass side streams. Considering the unique and highly complex chemical composition of coniferous needle/greenery biomass, biorefinery strategies can be considered as prospective possibilities to address top segments of the bio-based value pyramid, addressing coniferous biomass side streams as a source of diverse chemical substances with applications as the replacement of fossil material-based chemicals, building blocks, food, and feed and applications as fine chemicals. This study reviews biorefinery methods for coniferous tree forestry biomass side streams, exploring the production of value-added products. Additionally, it discusses the potential for developing further biorefinery strategies to obtain products with enhanced value.

Nutritional and Physicochemical Properties of Wild Lingonberry (Vaccinium vitis-idaea L.)-Effects of Geographic Origin.

In recent years, much attention has been devoted to Vaccinium L. berries because of their substantial potential to be adapted for the development of innovative food and pharmaceutical applications. The accumulation of plant secondary metabolites is extremely dependent on climate and other environmental conditions. In order to increase the reliability of the findings, this study was conducted with samples collected in four regions in Northern Europe (Norway, Finland, Latvia, and Lithuania) and analyzed in a single laboratory using a standardized methodology. The study aims to provide a comprehensive understanding of the nutritional (biologically active compounds (phenolic (477-775 mg/100 g fw), anthocyanins (20-57 mg/100 g fw), pro-anthocyanidins (condensed tannins (141-269 mg/100 g fw)) and antioxidant activity in different systems (ABTS•+, FRAP). Physicochemical properties (acidity, soluble solids, color) of wild Vaccinium vitis-idaea L. were also evaluated. The results may contribute to the development of functional foods and nutraceuticals with potential health benefits in the future. To the best of our knowledge, this is the first comprehensive report on the evaluation of the biologically active compounds of wild lingonberries from different Northern European countries based on one laboratory's validated methods. The results indicated a geomorphological influence on the biochemical and physicochemical composition of wild Vaccinium vitis-idaea L. depending on their place of geographical origin.

Procyanidins from Cranberry Press Residues-Extraction Optimization, Purification and Characterization.

Procyanidins are a polyphenolic group that can be found in a variety of foods such as chocolate, tea, cranberries and others. Type A procyanidins can be found in a handful of sources and one of the richest sources are American cranberries. These compounds possess antioxidative, anticancer and anti-inflammatory activities and are most widely used as prevention for urinary tract infections. Cranberries are utilized for jam and juice production, and the latter produces industrial food waste press residues. Press residues contain free and bound procyanidins which can be extracted for use as nutraceuticals. In this study, the extraction of cranberry press residues has been optimized using RSM and the resulting extracts have been purified and fractionated. The obtained procyanidin fractions have been investigated for their antioxidative potential and analyzed using LC-ESI-FTICR-HRMS to determine individual procyanidins. The optimization showed that the optimal extraction can be conducted using acetone in a concentration of 53% without the addition of an acidifying agent. Strong correlation was observed for procyanidin contents and their antioxidative activity using DPPH, ABTS and FRAP methods. The purified fractions contained 78 individual (65 Type A) procyanidins with the degree of polymerization of up to 9.

Temperature has a major effect on the cuticular wax composition of bilberry (Vaccinium myrtillus L.) fruit.

Cuticle is the first layer protecting plants against external biotic and abiotic factors and is responsive to climatic factors as well as determined by genetic adaptations. In this study, the chemical composition of bilberry fruit cuticular wax was investigated through a latitudinal gradient from Latvia (56°N 24°E) through Finland (65°N 25°E) to northern Norway (69°N 18°E) in two seasons 2018 and 2019. Changes in the major cuticular wax compounds, including triterpenoids, fatty acids, alkanes, aldehydes, ketones, and primary alcohols, were detected by GC-MS analysis. Generally, a decreasing trend in the proportion of triterpenoids from southern to northern latitudes, accompanied with an increase in proportion of fatty acids, aldehydes, and alkanes, in bilberry fruit cuticular wax was observed. A correlation analysis between climatic factors with proportion of wax compounds indicated that temperature was the main factor affecting the cuticular wax composition in bilberries. A controlled phytotron experiment with southern and northern bilberry ecotypes confirmed the major effect of temperature on bilberry fruit cuticular wax load and composition. Elevated temperature increased wax load most in berries of northern ecotypes. The level of triterpenoids was higher, while levels of fatty acids and alkanes were lower, in wax of bilberry fruits ripened at 18°C compared to 12°C in both northern and southern ecotypes. Based on our results, it can be postulated that the predicted increase in temperature due to climate change leads to alterations in fruit cuticular wax load and composition. In northern ecotypes, the alterations were especially evident.

Geographic Variability of Biologically Active Compounds, Antioxidant Activity and Physico-Chemical Properties in Wild Bilberries (Vaccinium myrtillus L.).

The aim of this study was to characterize the variation in biologically active compounds, antioxidant activity and physico-chemical properties in naturally grown bilberries gathered from different sites in Northern Europe. The variability in the biologically active compounds, antioxidant capacity and physico-chemical properties, as well as the development of tools for the authenticity and quality control of wild bilberries (V. myrtillus L.) in different geographical locations was evaluated. The berries of bilberries were handpicked during the summers of 2019 and 2020 during the time periods when they are typically harvested for commercial purposes in Northern Europe (Norway (NOR), Finland (FIN), Latvia (LVA) and Lithuania (LTU)). Berries from locations in NOR were distinguished by their higher mean TPC (791 mg/100 g FW, average), whereas the mean TPC of samples from the most southern country, LTU, was the lowest (587 mg/100 g FW). The TPC of bilberries ranged from 452 to 902 mg/100 g FW. The TAC values of investigated bilberry samples varied from 233 to 476 mg/100 g FW. A high positive correlation was found between TPC and antioxidant activity of the bilberry samples (R = 0.88 and 0.91 (FRAP and ABTS assays, respectively)), whereas the correlation between TAC and antioxidant activity was lower (R = 0.65 and 0.60). There were variations in the TPC and TAC values of investigated berries, suggesting that genotype also affects the TPC and TAC in berries. In 2020, the pH values and TSS contents of berries were significantly lower than in 2019. To the best of our knowledge, this is the first comprehensive reported evaluation of the biologically active compounds in wild bilberries from different Northern European countries using one laboratory-validated method.

Authentication of berries and berry-based food products.

Berries represent one of the most important and high-valued group of modern-day health-beneficial "superfoods" whose dietary consumption has been recognized to be beneficial for human health for a long time. In addition to being delicious, berries are rich in nutrients, vitamins, and several bioactive compounds, including carotenoids, flavonoids, phenolic acids, and hydrolysable tannins. However, due to their high value, berries and berry-based products are often subject to fraudulent adulteration, commonly for economical gain, but also unintentionally due to misidentification of species. Deliberate adulteration often comprises the substitution of high-value berries with lower value counterparts and mislabeling of product contents. As adulteration is deceptive toward customers and presents a risk for public health, food authentication through different methods is applied as a countermeasure. Although many authentication methods have been developed in terms of fast, sensitive, reliable, and low-cost analysis and have been applied in the authentication of a myriad of food products and species, their application on berries and berry-based products is still limited. The present review provides an overview of the development and application of analytical chemistry methods, such as isotope ratio analysis, liquid and gas chromatography, spectroscopy, as well as DNA-based methods and electronic sensors, for the authentication of berries and berry-based food products. We provide an overview of the earlier use and recent advances of these methods, as well as discuss the advances and drawbacks related to their application.

Syntrophy of Crypthecodinium cohnii and immobilized Zymomonas mobilis for docosahexaenoic acid production from sucrose-containing substrates.

Marine heterotrophic dinoflagellate Crypthecodinium cohnii is an aerobic oleaginous microorganism that accumulates intracellular lipid with high content of 4,7,10,13,16,19-docosahexaenoic acid (DHA), a polyunsaturated ω-3 (22:6) fatty acid with multiple health benefits. C. cohnii can grow on glucose and ethanol, but not on sucrose or fructose. For conversion of sucrose-containing renewables to C. cohnii DHA, we investigated a syntrophic process, involving immobilized cells of ethanologenic bacterium Zymomonas mobilis for fermenting sucrose to ethanol. The non-respiring, NADH dehydrogenase-deficient Z. mobilis strain Zm6-ndh, with high ethanol yield both under anaerobic and aerobic conditions, was taken as the genetic background for inactivation of levansucrase (sacB). SacB mutation eliminated the levan-forming activity on sucrose. The double mutant Zm6-ndh-sacB cells were immobilized in Ca alginate, and applied for syntrophic conversion of sucrose to DHA of C. cohnii, either taking the ethanol-containing fermentation medium from the immobilized Z. mobilis for feeding to the C. cohnii fed-batch culture, or directly coculturing the immobilized Zm6-ndh-sacB with C. cohnii on sucrose. Both modes of cultivation produced C. cohnii CCMP 316 biomass with DHA content around 2-3 % of cell dry weight, corresponding to previously reported results for this strain on glucose.

Trace Element Concentration and Stable Isotope Ratio Analysis in Blueberries and Bilberries: A Tool for Quality and Authenticity Control.

Vaccinium genus berries-wild bilberries (Vaccinium myrtillus L.) and cultivated highbush blueberries (Vaccinium corymbosum L.)-are consumed worldwide, and their consumption has a trend of stable increase. Thus, considering their wide use in ethnomedicine, for juice and jam production, as functional food, as well as their use in preparations of extracts which have application potential in pharmaceutical and cosmetics industries, studies regarding the composition of these berries are of special importance. The aim of this study is to characterise the elemental and isotopic composition, as well as variation in element concentration in bilberries gathered from different sites in Northern Europe and in commercially available blueberry samples from across the World. Furthermore, our aim was to develop tools for authenticity and quality control of these berries. The elemental composition of berries was analysed using inductively coupled plasma with optical emission detection (ICP-OED), while isotope ratio mass spectrometry (IRMS) was used for the determination of isotope ratio values. The results demonstrated detectable differences between macro- and microelement values in bilberries. IRMS analysis of blueberries revealed significant differences in isotope ratios based on the place of origin, indicating the possibility to use this analytical method for authenticity testing. In none of the samples, pollution was detected, even though there were indications of different growth conditions and geochemical differences affecting bilberry composition.

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.

Cuticular Wax Composition of Wild and Cultivated Northern Berries.

The outer-most layer of plant surface, the cuticle, consists of epi- and intra-cuticular wax. It protects the plant from dehydration, extreme temperatures and UV radiation, as well as attacks from pests such as molds and bacteria. Berry cuticular waxes are studied to understand the metabolism character (factors affecting wax layer composition in different berry species) and increase the microbial resistance and shelf life of berries. The aim of this study was analysis of the surface wax composition of nine species of wild and cultivated berries from Northern Europe. Cuticular wax analysis were done using gas chromatography-mass spectrometry. A total of 59 different compounds were identified belonging to nine groups of compounds, namely, alkanes, phytosterols, alcohols, fatty acids, phenolic acids, ketones, aldehydes, esters and tocopherols. The analyzed blueberries had the highest amount of wax present on their surface (0.9 mg berry-1), triterpenoids were the main wax constituent in these berries, with up to 62% wax composition. Berry species and varieties were compared based on their surface wax composition-similarities were found between different blueberry varieties; however, other berries showed differences based on concentration and composition of cuticular wax.

Compositional and morphological analyses of wax in northern wild berry species.

Aerial surfaces of plants are covered by a waxy cuticle protecting plants from excessive water loss and UV light. In the present study, composition and morphology of cuticular waxes of northern wild berry species bilberry (Vaccinium myrtillus L.), lingonberry (V. vitis-idaea L.), bog bilberry (V. uliginosum L.) and crowberry (Empetrum nigrum L.) were investigated. Scanning electron microscopy (SEM) revealed differences in epicuticular wax morphology, and gas chromatography-mass spectrometry (GC-MS) analysis confirmed variation in chemical composition of cuticular waxes between the berry species. The dominant compounds in bilberry and lingonberry cuticular waxes were triterpenoids, while fatty acids and alkanes were the dominant ones in bog bilberry and crowberry, respectively. Wax extracted by supercritical fluid extraction (SFE) from industrial press cakes of bilberry and lingonberry contained linoleic acid and γ-linolenic acid as the dominant compounds. Furthermore, in vitro sun protection factor (SPF) of berry waxes depicted good UV-B absorbing capacities.