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ObjectivesThe aim of this paper is to assess the relationship between demographic and socioeconomic predictors and the unmet health needs of the older adult population in Serbia.Materials and methodsThe study is part of the Population Health Survey of Serbia, which was conducted in the period from October to December 2019 by the Institute for Public Health of Serbia “Dr. Milan Jovanović Batut” and the Ministry of Health of the Republic of Serbia. The research was conducted on a representative sample of Serbian residents in the form of a cross-sectional study. For the purposes of this research study, data on senior citizens, aged 65 and older, were used.ResultsMultivariate regression analysis of demographic characteristics that showed statistical significance in the univariate model as a whole explains between 4.2% (Cox & Snell R Square) and 5.9% (Nagelkerke R Square) of the variance of unmet health needs and correctly classifies 66.3% cases. Statistically significant demographic predictors were the region where the respondents live, level of education, and material condition. The results of the research show that the most dominant predictors of the unmet health needs of the older adult population are related to socioeconomic inequalities, financial reasons, and predictors related to the inaccessibility of health care.ConclusionThe results suggest that individual socioeconomic predictors have a great influence on the emergence of unmet health needs of the older adult population in Serbia. Every third older adult resident did not receive the necessary health care, most often due to financial constraints.

Despite its promising potential, the capabilities of Saccharomyces paradoxus in commercial winemaking are still unutilized and require further investigation. In this study, the effects of fermentation by a S. paradoxus strain P01-161 on the composition of Malvazija istarska white wine in two harvest years were investigated. A range of complementary standard and metabolomics analysis approaches were applied, including OIV methods for basic parameters; HPLC-DAD-RI for organic acids, glycerol, and proteins; UPLC/MS/MS for phenolic compounds; and GC/FID, GC/MS, and GC × GC/TOF-MS for volatile compounds. The harvest year exhibited a significant impact, but many distinctive traits of S. paradoxus versus S. cerevisiae control wines were consistent across the seasons. These included reductions in malic acid and certain phenols and pathogenesis-related proteins. Saccharomyces paradoxus fermentation yielded higher levels of glycerol, volatile acidity, and specific thaumatin-like proteins. Among a total of 474 identified volatile compounds, S. paradoxus exhibited lower concentrations of several odoriferous alcohols, acids, and esters, as well as higher concentrations of β-damascenone, acetaldehyde, isobutyric acid, ethyl 2-methylbutyrate, ethyl acetate, isobutyl acetate, various esters of succinic and lactic acids, accompanied by numerous minor compounds, when compared to S. cerevisiae. These differences suggest the potential for distinct sensory profiles produced by the two yeasts, indicating that S. paradoxus could be a promising alternative for white wine production.

This study aimed to evaluate bioactive properties of Teran red wine by-products (grape skins, seeds, and wine lees) from six vinification treatments, including a control (7-day standard maceration). Pre-fermentative cryomaceration (8 °C; 48 h) and hot maceration (50 °C; 48 h), followed by the 13-day (CS15; C15; H15) and 28-day (C30; H30) period, considering fermentation/maceration and extended post-fermentative maceration, were conducted. In CS15, the saignée procedure was applied before fermentation/maceration. After maceration, the separation of by-products was performed, followed by lyophilization and solid–liquid extraction. Then, individual phenols were analyzed using high-performance liquid chromatography (HPLC), and total phenolic content (TPC) and antioxidant activity (FRAP) were analyzed using UV/Vis spectrophotometry. The results showed grape skins and wine lees in all treatments had significantly increased TPC and FRAP values compared to the control. The highest concentration of total phenols (HPLC) in grape skins was found in CS15, at 978.54 mg/100 g DW. In wine lees, the highest concentration of total phenols was detected in the 30-day maceration treatments, at 582.04 mg/100 g DW in C30, and 595.83 mg/100 g DW in H30, despite the pre-fermentative procedure. In grape seeds, the highest concentration of total phenols was found in the control (K7), at 432.42 mg/100 g DW. Pre-fermentative heating together with extended 30-day maceration (H30) strongly reduced the total levels of phenols (HPLC and TPC) in grape seed samples. The findings implied an evident impact of pre- and post-fermentative technologies on phenols and antioxidant activity in wine by-products of cv. Teran (Vitis vinifera L.).

The yeast Lachancea thermotolerans has the ability to produce notable amounts of lactic acid and reduce alcoholic strength in fermentation, so it has a considerable potential for mitigating negative impacts of climate changes in winemaking. In this study, a treatment with L. thermotolerans and Saccharomyces cerevisiae in sequential inoculation was compared to a control S. cerevisiae monoculture fermentation of Malvazija istarska (aka Malvasia Istriana) white grape must. Standard physico-chemical parameters of the obtained wines were determined by the OIV methods. Targeted (GC/FID and GC/MS) and untargeted (GC×GC/TOF-MS) gas chromatographic techniques were combined for the analysis of volatile compounds. Phenolic compounds were analyzed by UPLC/QqQ-MS/MS, and proteins by RP-HPLC-DAD, while a sensory analysis of wines was performed by a panel of trained and certified tasters. L. thermotolerans co-fermentation treatment increased the concentration of lactic acid and decreased alcoholic strength. L. thermotolerans increased the concentrations of geraniol, β-ionone, isobutanol, isobutyric acid, ethyl isobutyrate, several major acetates, ethyl lactate, and diethyl succinate, followed by many minor compounds. This wine also contained more hydroxycinnamoyl tartrates, while control S. cerevisiae wine had higher levels of free hydroxycinnamates. The effects on PR proteins were minor. L. thermotolerans co-fermentation slightly enhanced the sensory perception of tropical fruit, herbaceous, tobacco, and buttery odor notes, as well as fullness of body. With the largest number of identified volatile compounds up to date and other results obtained, this study contributes to the better understanding of oenological and especially aromatic potential of L. thermotolerans in white wine production.

This study investigates the effect of prolonged maceration, pre-fermentation heating, and barrel aging on the volatile aroma profile and sensory characteristics of Teran wine. The vinification processes included a control treatment (7-day maceration, TM7-Y; Y—young wine), 10-day maceration (TM10-Y), 21-day post-fermentation maceration (TM21-Y), and 48-h pre-fermentation heating at 45 °C followed by 8-day maceration (TPHT-Y). All wines were then aged in oak barrels for six months, resulting in TM7-A, TM10-A, TM21-A, and TPHT-A wines (A—aged wine). Volatile compounds were extracted using headspace solid-phase microextraction (HS-SPME) and analyzed by gas chromatography/mass spectrometry (GC/MS), while sensory profiles were evaluated using quantitative descriptive analysis (QDA). TPHT-Y and TM21-Y treatments reduced several groups of free volatile compounds while enhancing sensory properties, with TM21-Y wines notably exhibiting pronounced dried fruit notes, likely due to high β-damascenone concentrations. Conversely, TM10-Y and TM7-Y treatments resulted in significantly higher concentrations of the most volatile aroma compounds. Aging in oak barrels significantly increased the levels of particular free volatile compounds like C13-norisoprenoids, volatile phenols, furans, and lactones. It also enhanced sensory quality, with fruity aromas prominent across all treatments, and TM21-A and TPHT-A wines showing strong dried fruit, jam, and liqueur notes. This study offers valuable insights into tailoring wine aromas and sensory attributes through specific vinification techniques, contributing to a more refined approach to optimizing wine production. In conclusion, the findings highlight the importance of maceration and aging techniques in developing complex and desirable wine profiles, offering practical guidance for improving Teran wine quality.

It is not yet clear how adding silicon foliar fertilisation affects olive leaf (OL) phenolics and their potential to impact different cancer cells. Thus, we conducted a field trial to study the effect of foliar Si biostimulant fertilisation on the OL phenolic content of the ‘Leccino’ (LE) and ‘Istarska Bjelica’ (IB) cultivars. The experiment compared untreated Control (C) and three distinct levels of silicon (Si1, Si2, Si3) with Si concentrations of 0.55 g/L, 1.1 g/L, and 2.2 g/L, respectively. Si3 application resulted in the highest levels of oleuropein, apigenin-7-O-glucoside, luteolin-4-O-glucoside, rutin, and tyrosol compared to the C treatment. The polyphenols showed high cytotoxic activity in three cancer cell lines tested: cervical adenocarcinoma (HeLa), colon cancer (HCT116), and osteosarcoma (U2OS). The strongest inhibition of cell growth was observed in the HCT116 cell line. All cancer cells tested were more sensitive to treatment with polyphenols isolated from plants with added Si than those without added Si. The cytotoxic activity of the extracts on the healthy cell line RPE1 was similar to that on the cancer cell line HCT116 and U2OS.

In order to investigate the relationship between the occurrence of the “grubby” sensory defect caused by olive fruit fly (Bactrocera oleae (Rossi)) infestation and the resulting volatile composition, virgin olive oils were extracted from olives of the Leccino cultivar with 0%, 50%, and 100% olive fly infestations and subjected to analysis of the basic chemical quality parameters, fatty acids and volatiles, and sensory analysis by the Panel test. A 100% olive fly infestation reduced the basic chemical quality of the oil, while the fatty acid composition was not affected in any case. The overall sensory quality score and intensity of the positive sensory attributes decreased, while the intensity of the “grubby” defect increased proportionally to the degree of infestation. The occurrence and intensity of this defect were clearly causally related to the concentrations of 3-methylbutanal, 2-methylbutanal, β-ocimene, ethyl 2-methylbutyrate, dimethyl sulfoxide, 4-methyl-5H-furan-2-one, α-farnesene, 6-methyl-5-hepten-2-one, 1-octanol, E-2-nonen-1-ol, benzeneacetaldehyde, heptanal, and octanal, implying that the perception of “grubby“ comes from their joint contribution to the overall olive oil flavour. In addition to contributing to the understanding of the chemical origin of “grubby”, the results obtained could potentially be used to develop strategies to support sensory analysis in the classification of olive oil quality and the confirmation of the presence of this sensory defect in oil samples.

To investigate the effects of the application of bentonite, tannins, and their combination in alcoholic fermentation, Malvazija istarska (Vitis vinifera L.) white grape must was treated with 95 g/L of bentonite, 25 g/L of a hydrolysable tannin preparation, while the third treatment received the aforementioned doses of both agents. Control grape must was fermented without bentonite and exogenous tannins. All of the produced wines were additionally fined after fermentation with doses of bentonite needed to achieve complete protein stability. Wines were analyzed both after fermentation and after additional bentonite fining. Standard physicochemical parameters were determined by the OIV methods, and phenols were analyzed by high-performance liquid chromatography with diode-array detection (HPLC-DAD), while the concentrations of free and bound volatile aroma compounds were obtained after solid-phase extraction (SPE) followed by gas chromatography–mass spectrometry (GC-MS). Bentonite and tannins in fermentation generally reduced the total dose of bentonite needed for complete stabilization. Treatments with bentonite slightly decreased the concentration of total dry extract, while tannins preserved total acidity. The negative effect of bentonite on flavonoids was more severe. Tannins in fermentation preserved more hydroxycinnamoyltartaric acids with respect to control wine, and this effect was additionally enhanced by bentonite. Volatile and bound aroma composition was affected by all the treatments, while the addition of tannins resulted in higher concentrations of several important odoriferous esters, such as ethyl hexanoate, ethyl decanoate, and hexyl acetate. Additional fining with bentonite to complete protein stabilization annulled some of the positive effects observed after fermentation.

Six different vinification treatments, including a control treatment (7-day standard maceration) (K7), were performed to study the effects of non-standard techniques on bioactive compounds and sensory attributes of Teran red wine. Pre-fermentative mash cooling (8 °C; 48 h) and heating (50 °C; 48 h) followed by prolonged post-fermentative maceration of 13 days (C15;H15) or 28 days (C30;H30) were applied. In another treatment, after cooling, saignée was performed followed by 13-day prolonged maceration (CS15). Wine phenols and vitamins were analyzed by HPLC-DAD-FLD, minerals by ICP-OES, and sensory analysis was performed using the QDA and 100-point O.I.V./U.I.O.E. methods. Obtained results showed total phenolic concentration was the highest in the H30 treatment. The concentration of anthocyanins, flavan-3-ols and phenolic acids was significantly higher in wines of all vinification techniques compared to the control. Stilbene content was highly affected by pre-fermentative heating. Treatments CS15, H15, C30 and H30 resulted in the highest scores by both the QDA and 100-point sensory methods. The obtained results suggest that advanced non-standard vinification techniques have a significant impact on Teran wine by enhancing its composition of bioactive compounds and improving its sensory profile, which gives it an additional market value. Furthermore, a comprehensive comparison of such techniques applied simultaneously in one study is of substantial importance for additional research in wine production.

Research background. Baranjski kulen is one of the most popular fermented meat sausages originating from Croatia. It has protected geographical indication, and is traditionally produced in the Baranja region of Croatia. Kulenova seka is a fermented sausage very similar to baranjski kulen, but it has a different calibre and consequently, a shorter time of production. In recent decades, due to the high demand and popularity of these products, industrially produced baranjski kulen and kulenova seka have become available on the market. This work aims to identify specific characteristics of traditional and industrial sausages baranjski kulen and kulenova seka on proteome, peptidome and metabolome level, which could potentially lead to better optimization of the industrial production process in order to obtain an equivalent to the traditional product. Experimental approach. Protein profiles of baranjski kulen and kulenova seka (traditional and industrial) were analysed using two-dimensional gel electrophoresis followed by differential display analysis and protein identification by mass spectrometry. Peptidomic profile was analysed via liquid chromatography-tandem mass spectrometry. Furthermore, aroma profiles were investigated via headspace solid phase microextraction and gas chromatography-mass spectrometry. Results and conclusions. The major identified characteristics of each product were: industrial baranjski kulen: specific degradation of myosin-1 and titin, overabundance of stress-related proteins and increased phenylalanine degradation; traditional baranjski kulen: decreased concentration of phenylalanine and overabundance of fructose-bisphosphate aldolase A and carbonic anhydrase 3; industrial kulenova seka - specific myosin-4 and haemoglobin subunit alpha degradation process; traditional kulenova seka - overabundance of dihydropyrimidine dehydrogenase [NADP(+)] and myosin light chain 1/3, skeletal muscle isoform, degradation of albumin and myoglobin, decreased concentrations of almost all free amino acids and increased amounts of smoke-derived volatile compounds. Presented results showed that potential product type-specific quality markers for each sausage could be found. Novelty and scientific contribution. In this preliminary communication, the first insights into protein degradation processes and generation of peptides, free amino acids and aroma compounds of industrial and traditional baranjski kulen and kulenova seka are presented. Although further research is needed to draw general conclusions, the specific profile of proteins, peptides, amino acids and volatile compounds represents the first step in the industrial production of sausages that meet the characteristics of traditional flavour.

The majority of chemical compounds that contribute to varietal aroma originate from grape skin. To investigate the differences between volatile aroma compounds when different maceration conditions are applied, a total of six vinification treatments were carried out on Malvazija istarska (Vitis vinifera L.) variety, non-maceration control treatment (C), pre-fermentative two days cryomaceration treatment at 8 °C (CRYO), seven days maceration treatment at 16 °C (M7), 14 days maceration treatment at 16 °C (M14), and prolonged post-fermentative maceration treatments at 16 °C for 21 day (M21) and 42 days (M42). Wines were subjected to GC/MS and sensory analysis. Obtained results showed that prolonged post-fermentative maceration treatments contained the highest concentration of total volatile aroma compounds, precisely monoterpenes, alcohols, and other esters. Contrary, C and CRYO wines resulted in highest concentration of ethyl and acetate esters, and fatty acids. In addition, sensory analysis showed that longer maceration treatment wines (M14, M21, M42) were characterized by more aroma complexity, varietal flowery typicity, pronounced fruitiness, with accentuated dried fruit, moderate honey, and herbal notes. Obtained results can provide valuable information to producers when choosing an appropriate vinification technique based on the desired wine style which may lead to a further diversification of white wine market.

Malolactic fermentation (MLF) is a complex biochemical process playing an important role in the production of red wines. The main reasons for its implementation are the reduction of titratable acidity and the consequent increase of pH, microbiological stability of wine, and changes in aromatic and sensory properties of wine. The aim of this study was to determine the compatibility of yeast and bacteria used by different malolactic fermentation techniques and their influence on the fermentation duration, concentration of individual organic acids, aroma compounds, and on chemical and sensory properties of Teran wines. The experiment included control treatment (without MLF), spontaneous MLF, induced MLF at the beginning of alcoholic fermentation with simultaneous inoculation of yeast and bacteria (co-inoculation) and induced MLF after alcoholic fermentation (sequential MLF). In the co-inoculation treatment MLF had no negative effect on the alcoholic fermentation kinetic. Alcoholic fermentation was complete in all treatments. Co-inoculation resulted in a significantly shorter duration of the MLF process. In all MLF treatments, significant reduction of titratable acidity and the increase of pH values was noted. Wines of the spontaneous MLF treatment obtained the highest concentration of volatile acidity and ethyl acetate. In all MLF treatments a complete consumption of malic acid and a decrease in concentration of citric acid, total amount of higher alcohols, and acetaldehyde were observed. Furthermore, significantly higher concentrations of ethyl esters, diacetyl, acetoin, and 2,3-butanediol were present in wines from all MLF treatments.

For the first time the effects of different sampling periods and their interaction with five major autochthonous Croatian Istrian olive cultivars and the Italian cultivar ‘Leccino’ on the quantity and composition of olive leaf phenolic compounds and mineral nutrients were investigated. For that purpose, olive leaves were sampled in two collecting periods, in October and March, coinciding with the harvesting and pruning periods, respectively. All selected cultivars had a higher oleuropein leaf content in the pruning collecting period, with the highest levels noted for the ‘Leccino’ and ‘Buža’ cultivars. Cultivar significantly affected almost all the investigated phenols, with higher concentrations of these valuable compounds in the pruning than in the harvesting period. Differences observed in leaf mineral composition were closely related to the differences in phenolic profiles and were significantly affected by genotype. Some of the studied mineral nutrients, such as P, Cu and B, were found to be significantly correlated with the most abundant olive leaf phenolic compounds, oleuropein and verbascoside.

To investigate in detail the volatilomes of various Brassicaceae species, landraces, and accessions, and to extract specific volatile markers, volatile aroma compounds were isolated from plant samples by headspace solid-phase microextraction and analyzed by gas chromatography/mass spectrometry (HS-SPME-GC/MS). The data obtained were subjected to uni- and multivariate statistical analysis. In general, two cabbage (Brassica oleracea L. var. capitata) landraces emitted the lowest amounts of volatiles generated in the lipoxygenase (LOX) pathway. Wild species Brassica incana Ten. and Brassica mollis Vis. were characterized by relatively high trans-2-hexenal/cis-3-hexen-1-ol ratio in relation to other investigated samples. A Savoy cabbage (Brassica oleracea L. var. sabauda) cultivar and three kale (Brassica oleracea L. var. acephala) accessions exhibited particular similarities in the composition of LOX volatiles, while the LOX volatilome fraction of B. incana and B. mollis partially coincided with that of another wild species, Diplotaxis tenuifolia L. Regarding volatiles formed in the glucosinolate (GSL) pathway, Savoy cabbage and wild species B. incana, B. mollis, and D. tenuifolia showed more intense emission of isothiocyanates than cabbage and kale. Diplotaxis tenuifolia showed a rather limited production of nitriles. The results of this study contribute to the general knowledge about volatile composition from various Brassicaceae species, which could be exploited for their better valorization. Future studies should focus on the influence of various environmental, cultivation, and post-harvest factors to obtain data with a higher level of applicability in practice.

To investigate the utility of comprehensive GC×GC gas chromatography metabolomics in finding varietal markers among volatile compounds in non-aromatic red wines, representative samples of the two most important Croatian monovarietal red wines, Plavac mali and Teran, were subjected to analysis by both conventional gas chromatography–mass spectrometry (GC-MS) and comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC×GC-TOF-MS) after headspace solid-phase microextraction (HS-SPME). GC-MS was useful for the determination of the basic volatile profile composed mainly of major esters and acids, followed by terpenes and C13-norisoprenoids. GC×GC-TOF-MS allowed the identification of 209 volatiles, among which 49 were significantly different across monovarietal wines. The compounds most characteristic for Teran were two theaspirane isomers, ethyl 3-(methylsulfanyl)propanoate, ethyl methyhexanoate, and ho-trienol, whereas Plavac mali stood out with higher concentrations of tridecane and a 2,3-butanediol isomer. The two monovarietal wines were successfully differentiated between each other by multivariate statistical methods mostly based on GC×GC-TOF-MS data. The presented approach that combines conventional GC-MS and advanced GC×GC-TOF-MS showed a great potential for tracking chemical markers of varietal origin and could be practically applied in managing wine production, quality and typicity, marketing, and protection from forgery.

Up to date, there are no reports on the effects of combined sulfur (S) and nitrogen (N) foliar application in olive orchards on volatile compounds and sensory characteristics of virgin olive oil (VOO). In this work, the effects of increasing the fertilizer S and N concentration on volatile compound composition and odor and taste attributes of monovarietal VOOs of Istarska bjelica and Leccino cultivars were investigated. The volatile compounds were analyzed by gas chromatography with flame ionization and mass spectrometric detection after isolation by solid-phase microextraction, while sensory attributes were assessed by a professional panel. In all the investigated VOO samples, aldehydes were the most represented group of compounds, with (E)-2-hexenal as the most abundant, followed by (Z)-3-hexenal and 1-penten-3-one. Significant differences in the volatile profiles and sensory attributes were observed between VOOs from different treatments, mainly attributable to the interactions between all the factors that were investigated: treatment, cultivar, and year. Although significant interactions were noted for sensory attributes as well, the score for overall quality was generally lower for Leccino VOOs and for VOOs from both cultivars that were obtained after the highest SN dose treatment. All the samples were graded by overall sensory scores that were higher than eight and were characterized by well-pronounced fruitiness and the absence of sensory defects. The results of this study showed that foliar application of S and N significantly affects the quality of VOO, confirming that such a practice can modulate the characteristic olive oil odor and taste attributes and thus possibly influence its acceptability and preference among consumers.

There is a lack of studies evaluating the metabolic contribution of non-Saccharomyces yeasts in early fermentation phases. This study aimed to investigate the volatile aroma profiles produced by various non-Saccharomyces yeasts just before sequential inoculation with Saccharomyces cerevisiae to provide an insight into the particular effects they induce at this stage. The grape must of Malvazija istarska was inoculated with monocultures of Torulaspora delbrueckii, Metschnikowia pulcherrima, Pichia kluyveri, Lachancea thermotolerans, and Schizosaccharomyces pombe, alongside a S. cerevisiae control. Eighty volatile compounds were quantified via headspace solid-phase microextraction and gas chromatography–mass spectrometry, and the data were statistically elaborated. Volatile profiles of non-Saccharomyces yeasts differed significantly from the S. cerevisiae control. Most treatments caused increases in linalool and β-damascenone, decreases in higher alcohols and fatty acids, and improved synthesis of odoriferous esters. Torulaspora delbrueckii and M. pulcherrima produced compounds not commonly found in S. cerevisiae fermented wines. Multivariate statistical analysis linked the investigated yeasts to specific, particularly abundant compounds. Future studies should explore to what degree these contributions persist after sequential inoculation with S. cerevisiae in diverse grape must matrices.

This study investigates the effect of industrial scale filtration of fresh monovarietal virgin olive oil from Buža and Istarska bjelica cultivars on their volatiles, total phenols and sensory characteristics, and compares the oil samples clarified by filtration with those clarified by natural sedimentation/decantation after six months of storage. Filtration had a different effect on volatiles from the oil samples obtained from different cultivars. In the oil from Buža cultivar immediately after filtration only the amount of (Z)-2-pentenol slightly increased, but in Istarska bjelica the oil filtration affected eight compounds (the amount of hexanal, (E)-2-pentenal, (Z)-3-hexenal, (Z)-2-pentenol and (Z)-3-hexen-1-ol increased, while of hexyl acetate, (E)-2-penten-1-ol and (E)-2-hexen-1-ol decreased). In fresh filtered oil from Buža cultivar a slight decrease of total phenols was observed, while in those from Istarska bjelica the decrease was sharp, causing a decrease in the pungency and bitterness. Sedimentation/decantation had advantages over oil filtration of both cultivars, due to improved effect on the preservation of the sensory profile and the level of total phenols. Tentative aroma profiles based on odorant series obtained from the odour activity values were compared to the actual olive oil sensory profiles. These results could have a high level of applications in the olive oil industry for the optimization of the technology for obtaining monovarietal virgin olive oil with preserved specific and typical sensory characteristics, but also may serve experts to choose an appropriate virgin olive oil clarification method prior to analysis of volatile compounds.

Moderate wine consumption is often associated with healthy lifestyle habits. The role of wine as a healthy drink is mainly due to its bioactive compounds, which differ according to various viticultural and enological factors. The aim of the present study was to observe the differences in bioactive compounds of white and red autochthonous Croatian wines, differing in terms of the grape variety and production technology. Our further aim was to explore the effect of their moderate consumption (200 mL per day) over the course of six weeks on some aspects of health in sixty-six healthy individuals. Participants were divided into eight groups depending on the wines consumed, while one group formed a non-consuming control group. Medical examination and laboratory tests were performed before the start and at the end of the consumption period. Systolic and diastolic blood pressure, total cholesterol, and LDL concentrations decreased. Additionally, an increase in HDL concentrations, and serotonin and dopamine levels, was observed. ALT, ALP, and GGT levels did not significantly increase in consumer groups, although alcohol concentration was relatively high in all the wines. Such results support the beneficial effects of wine-derived bioactive compounds on some health aspects resulting from moderate white and red wine consumption.

This study investigates the effect of combined sulfur (S) and nitrogen (N) foliar fertilization on leaf S and N concentration, as well as on the growth of olive fruit and on the quantity and quality of olive oil, obtained from two olive cultivars ‘Istarska bjelica’ and ‘Leccino’ in two consecutive years. S and N are some of the most important nutrients, and both play a crucial role in plant oil production. The here-reported fertilization program significantly increased S concentration in leaves without affecting N concentration, which led to an increase in fruit yield and improvement of all fruit morphological parameters. The best oil yield per tree was obtained under the treatment with the highest S/N dose. Oil quality was not affected by S and N supply, and this allowed us to classify all our oil samples as extra virgin (EVOO). Regarding the content of total phenols (TPC) and composition of fatty acid methyl esters (FAME), they remained unaltered under the applied treatments. All investigated fruit morphological parameters, as well as fruit and oil yield, were highly cultivar-dependent. ‘Istarska bjelica’ was characterized as a cultivar with higher fruit mass and pulp percentage, while its stone parameters were lower than those of ‘Leccino’. Consequently, the extraction oil yield obtained from ‘Istarska bjelica’ fruits was much higher. Moreover, environmental conditions had a great impact on fruit and oil quantity. The here-obtained results led us to the conclusion that supply of S and N can enhance oil production without affecting its nutritional quality, a finding that could generate large long-term effects on economic growth in the olive oil sector.

Olive oil is considered one of the most valuable vegetable oils and is highly appreciated by consumers for its specific and distinguishable taste and aroma, as well as its nutritional value. Sterols and triterpene diols are important carriers of bioactive properties of olive oil and are responsible for some of the beneficial effects of its consumption on human health, such as lowering serum LDL-cholesterol levels and significantly reducing the risk of cardiovascular diseases. The concentration of total sterols and the proportions of particular sterols and triterpene diols are among the parameters used to verify and prove the authenticity of olive oil in accordance with the EU and other countries’ regulations. Finally, their composition has been shown to have high discrimination potential for ensuring traceability with respect to variety, geographical origin, harvest date, and other factors. For these reasons, the research on sterols and triterpene diols in olive oil is an ever-growing field of scientific interest with great practical importance. This review focuses on all the important aspects of sterols and triterpene diols in olive oil, from their chemical structure, biosynthesis, occurrence and role in plants, health benefits, and their use in official controls of olive oil purity and authenticity, to a conclusive survey on the recent findings about the effects of different factors of influence on their content and composition, with a detailed comparative analysis of studies that investigated the effects of the two most important factors, variety and ripening degree.

The effect of the addition of different amounts of olive leaf (1, 2.5, and 5%, m/m) during Buža olive cv. oil production on the quantitative production parameters, composition, and sensory characteristics of the obtained oils were investigated in this study. The addition of leaf during oil extraction increased oil yield and extractability index by 97% compared to the control oil. The addition of leaf during extraction increased the concentration of pigments in oils, and the oil positive sensory attributes intensities, such as fruitiness and green grass/leaf notes. The influence on oil phenolic composition was dependent on the amount of leaf added. When 1% leaf was added, most of the phenolic compounds were preserved, while the addition of leaf at 5% decreased the concentration of the majority of phenols, especially secoiridoids by 45% compared to the control oil. The addition of leaf slightly increased the concentration of fatty acid ethyl esters and waxes in the oils. The obtained results indicate that particular importance should be given to the amount of olive leaf present in olive paste during oil extraction, since it apparently can increase the extractability of oil, but can also have negative effects on phenolic composition when added in excess.

Manganese (Mn) is an essential element that intervenes in several plant metabolic processes. The olive tree, and its fruits and leaves, are known as a source of nutraceuticals since they are rich in biophenols. However, there is still a serious lack of data about biophenolic distribution in olive stems and roots under Mn fertilisation. In this context, our study aimed to examine the effects of Mn fertilisation on the biophenolic profile in the leaves, stems, and roots of the ‘Istarska bjelica’ olive cultivar. The experiment was set up in a greenhouse, during a period of five months, as a random block design consisting of three treatments with varying Mn concentrations in full-strength Hoagland’s nutrient solution (0.2 µM Mn, 12 µM Mn, and 24 µM Mn). The obtained results indicate that the amount of Mn in the examined olive plant tissues was significantly higher under 12 µM Mn and 24 µM Mn treatments compared to that of the 0.2 µM Mn treatment. While the concentration of biophenols varied in roots depending on the compound in question, a strong positive impact of the increased Mn concentration in nutrient solution (12 µM Mn and 24 µM Mn) on the concentrations of the main biophenolic compounds was observed in stems. The concentration of oleuropein in leaves almost doubled at 24 µM Mn, with the highest Mn concentration, as compared to the 0.2 µM Mn treatment. The obtained results led to the conclusion that the supply of Mn could enhance the concentration of some biologically active compounds in olives grown hydroponically, implying a critical need for further investigation of Mn fertilisation practices in the conventional olive farming system.

To differentiate monovarietal wines made from native and introduced varieties in Istria (Croatia), samples of Malvazija istarska, Chardonnay and Muscat yellow from two harvest years (2013 and 2014) were subjected to headspace solid-phase microextraction and gas chromatographic/mass spectrometric analysis (HS-SPME-GC/MS) of volatile aroma compounds. Significant effects of variety and harvest year were determined, but their interaction complicated the differentiation. Particular compounds were consistent as markers of variety in both years: nerol for Malvazija, ethyl cinnamate and a tentatively identified isomer of dimethylbenzaldehyde for Chardonnay, and terpenes for Muscat yellow. Wines from 2013 contained higher concentrations of the majority of important volatiles. A 100 % correct differentiation of Malvazija istarska and Chardonnay wines according to both variety and harvest year was achieved by stepwise linear discriminant analysis.

To differentiate white wines from Croatian indigenous varieties, volatile aroma compounds were isolated by headspace solid-phase microextraction (HS-SPME) and analyzed by comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC×GC-TOF-MS) and conventional one-dimensional GC-MS. The data obtained were subjected to uni- and multivariate statistical analysis. The extra separation ability of the GC×GC second dimension provided additional in-depth volatile profile information, with more than 1000 compounds detected, while 350 were identified or tentatively identified in total by both techniques, which allowed highly efficient differentiation. A hundred and sixty one compounds in total were significantly different across monovarietal wines. Monoterpenic compounds, especially α-terpineol, followed by limonene and linalool, emerged as the most powerful differentiators, although particular compounds from other chemical classes were also shown to have notable discriminating ability. In general, Škrlet wine was the most abundant in monoterpenes, Malvazija istarska was dominant in terms of fermentation esters concentration, Pošip contained the highest levels of particular C13-norisoprenoids, benzenoids, acetates, and sulfur containing compounds, Kraljevina was characterized by the highest concentration of a tentatively identified terpene γ-dehydro-ar-himachalene, while Maraština wine did not have specific unambiguous markers. The presented approach could be practically applied to improve defining, understanding, managing, and marketing varietal typicity of monovarietal wines.

The interactive effects of cultivar, collecting period, and geographical location on the content and composition of biophenols and macro and micronutrients in olive (Olea europaea L.) leaf were investigated. Leaves of six cultivars were collected at three periods in two locations in Croatia. The leaves of Istarska bjelica cultivar had the greatest biophenol (oleuropein) potential, especially those sampled in January and in March at the location of Pag. All the cultivars yielded leaves with the highest concentration of biophenols in March, which coincided with the pruning period. Except for high oleuropein concentration in Istarska bjelica, flavonoids were found to be most useful for differentiating olive leaves according to cultivar. Verbascoside turned out to be the most potent differentiator of collecting periods, while phosphorus and zinc turned out to be most useful for differentiating locations. Despite different agroecological conditions at the two locations, cultivar exhibited a significant effect on olive leaf nutrient composition, which was certainly causally related to that of the biophenols. The results obtained showed that it is possible to plan more well-timed and efficient exploitation of biophenols from olive leaf based on the knowledge about the interactive effects of the three studied factors.

With the aim to investigate the influence of post-harvest olive fruit storage temperatures on virgin olive oil production parameters, composition and quality, Istarska bjelica (IB) and Rosinjola (RO) fruits were stored for seven days at room temperature (RT), +4 °C and −20 °C prior to oil production. Lower temperatures delayed post-harvest maturation of IB fruits. Theoretical oil content did not change depending on the storage temperature, while the highest oil yield and extractability index were obtained after storage at RT. Chlorophylls decreased in IB-RT and in IB-20. A decrease in the sensory quality of oils was detected after fruit storage at RT and −20 °C, while the refrigeration temperature of +4 °C preserved it. Regarding the content of fatty acid ethyl esters, an increase was observed in IB-RT oils. Storage at RT increased the content of waxes, while the lower temperatures partially suppressed this phenomenon. In oils of both cultivars, storage at +4 °C preserved the concentration of most phenolic compounds at a level more similar to that of the fresh oil when compared to the other two treatments. In the production conditions, when prolonged fruit storage is necessary, refrigeration seems to be the most suitable option.

To test the effect of the moment of bentonite addition, co-addition of tannins, and bentonite type on the differential affinity of pathogenesis-related (PR) proteins towards bentonite during grape must fermentation, three separate experiments were set up. PR proteins in the obtained wines were analyzed by reverse phase and size exclusion high-performance liquid chromatography (HPLC). The most significant reduction of bentonite dose and PR protein concentration was achieved by applying bentonite in the last third of fermentation. Particular thaumatin-like proteins (TLP) and proteins with lower molecular mass in general were more affected than others, while TLPs were more affected than chitinases. Exogenous enological tannins interacted with particular PR proteins, mostly TLPs, and lowered the total bentonite dose required. The combined application of tannins and bentonite in fermentation removed more PR proteins than bentonite alone, but did not achieve a synergistic effect in reducing the bentonite dose. Various bentonite types, including two Na-activated bentonites, an activated Na bentonite with specifically adsorbed silica, and an active Na-Ca bentonite, exhibited differential affinity towards different PR proteins. The results obtained could be used in developing wine fining protocols which combine treatments with complementary affinity for adsorption and removal of PR proteins, and in this way achieve greater efficiency of bentonite fining by reducing its total dose, which is of significant interest to the wine industry.

In order to investigate the potential of various olive cultivars and leaf sampling times for phytochemical farming practice in Croatia, phenolic and mineral composition was determined in olive leaves of four Croatian cultivars and Italian cultivar Leccino collected at three occasions, in October 2017, January 2018, and March 2018. Istarska bjelica turned out to have the largest phytochemical potential among the investigated cultivars due to steady high oleuropein concentrations found in its leaves. The concentration of main phenolic components in Istarska bjelica leaves changed only slightly during the sampling period, suggesting the possibility of its higher capability for low air temperatures stress resistance and different metabolic response compared to the other studied cultivars. Low air temperatures increased the oleuropein level and antioxidant activity in leaves of Leccino, Oblica, Levantinka, and Drobnica cultivars, which may be of crucial phytochemical farming interest. Each of the investigated olive cultivars was characterized by a specific leaf mineral nutrient composition, which could have had a specific role in their interplay with phenols.

To investigate the phenolic and aroma composition of Malvazija istarska (Vitis vinifera L.) white wines produced by an unconventional technology comprising prolonged maceration followed by maturation in wooden barrels, representative samples were subjected to analysis by UV/Vis spectrometry, high-performance liquid chromatography, and gas chromatography-mass spectrometry. When compared to standard wines, the investigated samples contained higher levels of dry extract, volatile acidity, lactic acid, phenols, colour intensity, antioxidant activity, majority of monoterpenes, C13-norisoprenoids, methanol, higher alcohols, ethyl acetate, branched-chain esters and esters of hydroxy and dicarboxylic acids, ethylphenols, furans, and acetals, as well as lower levels of malic acid, β-damascenone, straight-chain fatty acids, ethyl and acetate esters. It was estimated that maceration had a stronger influence on phenols, and maturation on volatile aromas. Despite different vintages and technological details, the investigated wines showed a relative homogeneity in the composition, representing a clear and distinctive type.

This interdisciplinary study aims to contribute to the characterization of Istrian (Croatia) olive cultivars and oil, giving for the first time the morphological and genetic profile of Bova cultivar, and chemical and sensorial characteristics of its oil. Morphological features of Bova cv. were determined according to the International Olive Council methodology, while molecular characterization was performed using eleven microsatellite markers. Bova cultivar was morphologically and genetically different from other described Istrian olive cultivars. The microsatellite profile of Bova was also unique when compared to more than 200 different Italian genotypes using the same set of markers. In order to characterize the oil from Bova cv., fruits from three trees at the same ripening stage were harvested and processed separately under the same conditions. Volatile composition of the obtained oil samples was determined using solid-phase microextraction with gas chromatography/mass spectrometry. About 50 volatiles were detected, mostly hydrocarbons (34.69 % of total peak area), followed by aldehydes (25.80 %), alcohols (22.24 %), ketones (8.76 %), organic acids (4.08 %), terpenes (2.10 %), esters (2.18 %) and furans (0.26 %). Bova oil was rich in total C6 (39.87 %) and C5 volatiles (13.85 %), biogenerated through the lipoxygenase pathway. The most prevalent volatile compound was C6 aldehyde E-2-hexenal. Quantitative descriptive sensory analysis of the investigated olive oil samples was carried out by a sensory panel. The sensory profile was characterized by medium intensities of olive fruity, green leaves and grass, light tomato and aromatic herbs flavours with mild apple and other ripe fruit notes. The taste was characterized by medium to strong bitterness, followed by medium pungency and mild sweetness.

Despite having an interesting native olive gene pool and a rapidly emerging olive oil industry, monovarietal extra virgin olive oils (EVOO) from Croatia are relatively unexplored. To investigate the inter-varietal diversity of typical volatile and phenolic profiles of Croatian EVOO, 93 samples from six olive (Olea europaea L.) varieties were subjected to gas chromatography-ion trap mass spectrometry (GC-IT-MS) and ultra-performance liquid chromatography with diode array detection (UPLC-DAD), respectively. Quantitative descriptive sensory analysis was also performed. Analysis of variance extracted many relevant exclusive or partial discriminators between monovarietal EVOOs among the identified volatile compounds and phenols. Successful differentiation model with a 100% correct classification was built by linear discriminant analysis, while the most typical volatiles for each monovarietal EVOO were confirmed by partial least squares discriminant analysis. Diverse typical sensory attributes among the EVOOs were tentatively ascribed to the variations in the composition of volatiles and phenols. It was proven that the approach that comprises GC-IT-MS and UPLC-DAD analysis may provide additional objective information about varietal origin and typicity which successfully complement those obtained by sensory analysis. The approach was characterized as universal in nature, with a significant potential to contribute in strengthening the varietal identities and position on the market of monovarietal and Protected Denomination of Origin (PDO) EVOO.

In order to differentiate the extra virgin olive oils (EVOO) of different origin of purchase, such as monovarietal Italian EVOO with protected denomination of origin (PDO) and commercial-blended EVOO purchased in supermarkets, a number of samples was subjected to the analysis of volatile aroma compounds by both targeted gas chromatography/mass spectrometry (GC-MS) and untargeted profiling by comprehensive two-dimensional gas chromatography/mass spectrometry (GC×GC-TOF-MS), analysis of phenols by targeted high-performance liquid chromatography/mass spectrometry (HPLC-DAD-ESI/MS), and quantitative descriptive sensory analysis. Monovarietal PDO EVOOs were characterized by notably higher amounts of positive LOX-derived C6 and C5 volatile compounds, which corresponded to the higher intensities of all the assessed positive fruity and green odor sensory attributes. Commercial-blended EVOOs had larger quantities of generally undesirable esters, alcohols, acids, and aldehydes, which coincided with the occurrence of sensory defects in many samples. Many minor volatile compounds that were identified by GC×GC-TOF-MS were found to differentiate each of the two investigated groups. The differences between the groups with respect to phenols and taste characteristics were evident, but less pronounced. The results that were obtained in this study have undoubtedly confirmed the existence of the large heterogeneity of oils that are sold declared as EVOO. It was shown that GC-MS, GC×GC-TOF-MS, and HPLC-DAD-ESI/MS analyses have complementary outputs, and that their use in combination has advantages in supporting the results of sensory analysis and objectively differentiating these groups of EVOO.

The volatile compounds found in virgin olive oil, mainly C6 and C5 volatile compounds biogenerated from polyunsaturated fatty acids through the lipoxygenase pathway, are responsible for their particular aroma. The composition of volatile compounds in olive oil depends on the cultivar, the ripening degree of the fruits and processing conditions. Among many different autochthonous cultivars in Istria (Croatia), some of the most prevalent are Buža, Črna and Rosinjola. The volatiles and sensory characteristics of their monovarietal virgin olive oil are little known. Therefore, fruits from these three cultivars were handpicked at the same ripening degree and processed under the same conditions. Quantitative descriptive sensory analysis of monovarietal virgin olive oil was carried out by the panel. Volatile composition was evaluated by headspace solid phase microextraction-gas chromatography, previously optimized and validated. The main parameters affecting effectiveness, time and temperature of extraction were optimized. The extraction procedure showed detection and quantification limits, as well as linear ranges adequate for the analysis of selected volatile compounds. Good precision was obtained both in terms of intra-day repeatability (relative standard deviations generally lower than 7 %) and inter-day precision. The tested types of monovarietal olive oil showed different volatile profiles, although E-2-hexenal was the main compound in all samples. Buža oil was the richest in total C6 and C5 volatile compounds. The results show that the most important contributors to the olive oil aroma (odour activity value >1.0) were 1-penten-3-one, E-2-hexenal, hexanal, hexanol, Z-3-hexen-1-ol and Z-2-penten-1-ol. These chemical findings were compared with those provided by the panel test. Buža had the highest intensity of sensory characteristic 'other ripe fruits' and Rosinjola had the highest intensity of sensory characteristic 'bitter'. All results show that olive oil aroma compounds accumulate differently depending on the cultivar, indicating a close dependence on the enzymatic pool, which is genetically dependent.

The influence of different grape mash maceration treatments on the varietal and secondary aroma profiles of wines produced from an aromatic red grape variety, Muškat ruža porečki, have been investigated. Two essentially different techniques were applied, fermentative maceration at room temperature and prefermentative cryomaceration, both in durations of one, three and five days. Generally, higher concentrations of free and bound varietal aroma compounds were found in wines obtained by maceration at room temperature in relation to cryomaceration. Regarding the effect of the duration of maceration, the highest concentrations were determined in wines obtained by three-day maceration treatments, in both fermentative and cryomaceration treatments. Secondary aroma compounds followed a less uniform pattern. The compounds with the highest odour unit values in all investigated wines were linalool, citronellol, geraniol, β-damascenone, β-ionone, isoamyl alcohol, 2-phenylethanol, ethyl hexanoate, ethyl octanoate, isoamyl acetate, ethyl acetate, and diethyl succinate. It has been shown that Muškat ruža porečki is an aromatic variety, producing wines with notable monoterpenol fraction, which are characterized by a typical varietal Muscat aroma with a dominant rose odour accompanied by red fruit nuances. Sensorially, longer maceration treatments improved odour and overall wine quality, together with the intensity and recognisability of varietal Muscat aroma, while short-term cryomaceration emerged as a preferable technique for the production of light rosé wines with pronounced Muscat aroma and low phenolic content. The presented maceration techniques were shown to be applicable for the production of different types of Muškat ruža porečki rosé and red wines.

In order to investigate the composition of secondary aroma compounds of fresh grape marc distillates as a result of variety and production technology, 30 samples (6 varieties×5 samples) were analysed. White grape marc samples from Malvazija istarska, Chardonnay and Muscat Blanc were obtained as by-products in standard white wine production, while red grape marc samples from Teran and Cabernet Sauvignon were obtained after standard red wine production procedures. Marc from red grape variety Muškat ruža porečki was obtained during the production of rosé wines. All fermented marc samples were distilled using a traditional copper alembic. The obtained distillates were subjected to GC/MS and GC/FID analyses. Malvazija istarska distillates exhibited exceptionally high methanol content. Distillates from white grape varieties were found to be characterized by higher C6 alcohol and 1-propanol concentrations, while red grape distillates contained higher amounts of the majority of alcohols, acids, and esters. In Muškat ruža distillates intermediate concentrations of many important aroma compounds were found. It was concluded that differences in the production technology parameters, depending on the variety, resulted in differences in secondary aroma profiles, most evident between distillates from white and red varieties. These findings were confirmed applying stepwise linear discriminant analysis (SLDA), which resulted in 100 % correct classification of distillates according to the variety and corresponding production technology.

During a two-year study, the effect of different grape mash maceration treatments (skin contact at 20 °C for 10, 20 and 30 h) and cryomaceration (skin contact at 7 °C for 10, 20 and 30 h) on the concentration of free and bound monoterpenes and total phenols in Malvazija istarska wine (the most spread white wine variety in Istria) was monitored. Monoterpenic alcohols linalool, a-terpineol, citronellol, nerol and geraniol were determined by gas chromatography after isolation on octadecylsilica (C18) sorbent and elution with pentane/dichlorometane ratio of 2:1, while total phenols were determined spectrophotometrically with Folin-Ciocalteau reagent. Sensory evaluation of wines was performed using the Buxbaum method. The obtained results show that linalool and geraniol are the most abundant monoterpenes in Malvazija istarska, and that grape mash maceration treatments cause significant increase of free and bound monoterpene concentrations compared to those in control treatment wine obtained without maceration. The content of phenols, which are subject to oxidation, was found to be significantly lower in wines produced by cryomaceration treatments, while in those produced by maceration at 20 °C the concentration of total phenols increased proportionally with the duration of maceration, negatively influencing wine quality by causing higher colour intensity and bitter taste.