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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.

Agriculture is facing challenges to produce more food in a climate scenario that works in the opposite direction. To amend this, agriculture has to invent new ways of making more with less. Interest in using by-products and finding new ways to utilize them has been increasing in recent years. The use of protein-rich sources for protein hydrolyzation and the use of these protein hydrolysates as biostimulants in plant production have been increasing. These mixtures are mainly produced by chemical and enzymatic hydrolysis from agro-industrial protein-rich by-products of animal, plant, and algal origins. The application of PHs has the potential to alleviate environmental stress; improve plant growth; and increase productivity, fruit yield, and abiotic-stress tolerance in agricultural crops. The use of these biostimulants offers a way to reduce the use of agrochemicals and agrees with the “do more with less” task in the future of agricultural production. This review gives an insight into the production of PHs, referring to sources of raw materials and methods of hydrolysis, the uptake and translocation of PHs, their effect on plant growth, the development and physiology, their role in alleviating stressful conditions, and their use in agriculture. The beneficial effects of PHs on different aspects of plant physiology, metabolism, and plant functioning under stressful conditions are evident. Inconveniently, crops, and sometimes even cultivars, are affected differently based on the way that PH is applied, the timing, and the concentration applied. Further research is needed to elucidate the mechanisms by which the components of PHs modify plant physiology and metabolism.

Objectives: The aim of this study was to determine the antifungal activity of olive leaf extract (OLE) and the synergistic effect of standard antifungal therapy and OLE against clinical oral Candida species’ isolates. Materials and Method: The susceptibility of 60 clinical isolates of the Candida species (36 C. albicans, 16 C. krusei, 5 C. glabrata and 3 C. tropicalis) was tested with four concentrations of OLE (60 µg/µL, 120 µg/µL, 240 µg/µL and 333 µg/µL) and the synergistic effect of standard antifungal therapy and OLE (miconazole (MIC) + 333 µg/µL OLE and nystatin (NYS) + 333 µg/µL OLE). The antimicrobial activity was tested using the disk diffusion method. Results: All concentrations (60 µg/µL, 120 µg/µL, 240 µg/µL and 333 µg/µL) of OLE showed a statistically significant effect on all Candida species compared to the control (DMSO) except for the lowest concentration (60 µg/µL) tested on C. glabrata. There was a dose-dependent effect of OLE on tested samples. Concentrations of 240 µg/µL and 333 µg/µL showed statistically significant higher antifungal activity compared to the lowest concentration of 60 µg/µL. No statistically significant synergistic effect of OLE and standard antifungal therapy was found compared with standard therapy alone. Conclusions: The results of this study present the significant antimicrobial effect of OLE against all tested Candida species except for the lowest concentration on C. glabrata. Increasing the concentration of OLE also increases its effect on Candida species. This indicates the possible potential effect of OLE in the treatment of Candida-related oral diseases.

This study explores the potential of olive leaves, long integral to Mediterranean traditional medicine, as a rich source of valuable compounds. The challenge lies in their considerable water content, hindering these compounds’ full valorization. Four drying methods (air-drying, oven-drying, freeze-drying and solar-drying) were investigated for their impact on nutrient and bioactive compound content in the leaves of four olive varieties (“Arbequina”, “Koroneiki”, “Menara” and “Picholine Marocaine”) cultivated in Morocco. In their fresh state, “Picholine Marocaine” exhibited the highest protein levels (6.11%), “Arbequina” had the highest phenolic content (20.18 mg gallic acid equivalents/g fresh weight (FW)), and “Koroneiki” and “Menara” were highest in flavonoids (3.28 mg quercetin equivalents/g FW). Specific drying methods proved optimal for different varieties. Oven-drying at 60 °C and 70 °C effectively preserved protein, while phenolic content varied with drying conditions. Air-drying and freeze-drying demonstrated effectiveness for flavonoids. In addition, an analytical approach using high-performance liquid chromatography and diode array detection (HPLC-DAD) was applied to investigate the effects of the different drying methods on the bioactive fraction of the analyzed samples. The results showed qualitative and quantitative differences depending on both the variety and the drying method used. A total of 11 phenolic compounds were tentatively identified, with oleuropein being the most abundant in all the samples analyzed. The freeze-dried samples showed the highest content of oleuropein in the varieties “Arbequina” and “Picholine Marocaine” compared to the other methods analyzed. In contrast, “Koroneiki” and “Menara” had higher oleuropein content when air dried. Overall, the obtained results highlight the importance of tailored drying techniques for the preservation of nutrients and bioactive compounds in olive leaves.

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.

The lipoxygenase pathway has a significant influence on the composition of the volatile components of virgin olive oil (VOO). In this work, the influence of the maturity index (MI) on the activity of the lipoxygenase enzyme (LOX) in the fruits of the autochthonous Dalmatian olive cultivars Oblica, Levantinka and Lastovka was studied. The analysis of the primary oxidation products of linoleic acid in the studied cultivars showed that LOX synthesises a mixture of 9- and 13-hydroperoxides of octadecenoic acid in a ratio of about 1:2, which makes it a non-traditional plant LOX. By processing the fruits of MI~3, we obtained VOOs with the highest concentration of desirable C6 volatile compounds among the cultivars studied. We confirmed a positive correlation between MI, the enzyme activity LOX and the concentration of hexyl acetate and hexanol in cultivars Oblica and Lastovka, while no positive correlation with hexanol was observed in the cultivar Levantinka. A significant negative correlation was found between total phenolic compounds in VOO and LOX enzyme activity, followed by an increase in the MI of fruits. This article contributes to the selection of the optimal harvest time for the production of VOOs with the desired aromatic properties and to the knowledge of the varietal characteristics of VOOs.

Olive groves in the Mediterranean may lose production sustainability because of their vulnerability to climatic change. Irrigation is an important measure that could significantly affect fruit yield, olive fruit fly infestation, and oil characteristics. The aim of paper was to compare the regulated deficit irrigation with different water management practices, in consecutive years, in two locations in Zadar County (Croatia), affecting fruit morphology, olive fruit fly infestation, and quantity and quality of the extracted Coratina cultivar oil. Treatments, namely C—rainfed, T1—deficit irrigation (produce’s practice), T2—regulated deficit irrigation, and T3—full irrigation (100% ECTO), were established. Irrigated treatments had a positive effect on all morphological characteristics of the fruit. The pulp mass, independently of the year, increased in irrigated treatment (ranging from 1.04 to 1.65 in C to 2.25 and 2.30 in the irrigated treatments) and resulted in a higher oil content on a fresh weight basis (ranging from 16.39% to 17.85% in C to 19.48% to 23.26% in the irrigated treatments). However, fruit yield per tree was only location-dependent. When olive fruit fly presence was high, fruit infestation was greatest in the irrigated compared to the rainfed treatment. According to quality parameters, all oils were classified as EVOO. Individual phenols were influenced by irrigation, while the composition of fatty acids was more influenced by location than treatment. The sensory characteristics of the resulting oil were slightly reduced compared to rainfed treatment. The results indicate that regulated deficit irrigation benefits water use sustainability without compromising the quality of the oil.

In this study, we investigated the influence of pedological parameters and variation of altitude on the mineral nutrients, phenolic compounds, and antioxidant activities of olive leaves. Samples of the Chetoui cultivar were collected from eight geographical locations with different altitudes. Levels of phenolic compounds varied according to the altitude. Classification of the locations revealed that altitude 1 (>500 m) was characterized by high levels of secoiridoids and simple phenols, while altitude 2 (500–300 m) and altitude 3 (O glucoside, the most important phenols in Chetoui olive leaves. The results suggest that, in addition to pedological criteria, environmental conditions also influence the formation of phenolic compounds.

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.

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 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.

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.

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.

Organic amendments are often reported to improve soil properties, promote plant growth, and improve crop yield. This study aimed to investigate the effects of the biochar and compost produced from sewage sludge and olive pomace on soil hydraulic properties, water flow, and P transport (i.e., sorption) using numerical modeling (HYDRUS-1D) applied to two soil types (Terra Rosa and Rendzina). Evaporation and leaching experiments on soil cores and repacked soil columns were performed to determine the soil water retention, hydraulic conductivity, P leaching potential, and P sorption capacity of these mixtures. In the majority of treatments, the soil water retention showed a small increase compared to the control soil. A reliable fit with the modified van Genuchten model was found, which was also confirmed by water flow modeling of leaching experiments (R2 0.99). The results showed a high P sorption in all the treatments (Kd 21.24 to 53.68 cm3 g−1), and a high model reliability when the inverse modeling procedure was used (R2 0.93–0.99). Overall, adding sewage sludge or olive pomace as compost or biochar improved the Terra Rosa and Rendzina water retention and did not increase the P mobility in these soils, proving to be a sustainable source of carbon and P-rich materials.

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.

Food safety is one of the main concerns in the world and in wine it depends mostly on metabolites of microbial origin. The aim of this study was to investigate the occurrence of natural contaminants, ochratoxin A and biogenic amines (cadaverine, histamine, putrescine and tyramine), in Croatian commercial red wines originating from different Croatian wine-making regions. Ochratoxin A was detected in 92.8% of samples, however its concentrations in all samples were more than 10-fold lower than the limit set by the European Union (2 µg/kg), marking these wines as safe for consumption. The frequency of occurrence and measured concentrations of ochratoxin A were higher in wines produced in southern regions with highest values obtained in wines from southern Dalmatian islands. All samples were contaminated with cadaverine and putrescine, while 88.2% and 82.7% were contaminated with histamine and tyramine, respectively. Histamine concentrations ranged from below the limit of detection to 8.5 mg/L. Higher histamine concentrations were measured in wines with higher pH values which coincided with southern geographic origin. These results reinforce the need for routine detection and quantification of biogenic amines in Croatian wines to achieve better control of vinification and minimize their formation.

Oral infections caused by Candida species are becoming more common, which may be related to an increase in the number of immunologically compromised patients as well as favorable conditions in the oral cavity that often include removable prosthetic appliances. The purpose of this study was to determine the presence of a particular Candida species in patients with PMMA and Cr-Co prosthetic appliances, as well as the salivary flow rate, and oral signs and symptoms. This investigation included a total of 120 subjects with different kinds of removable dentures. A sample of concentrated oral rinse was collected from all subjects in order to detect Candida colonization and identify the Candida species, a quantum of salivation was measured, and subjects were examined clinically. Candida spp. was predominant among the subjects who were denture wearers (p < 0.0001). In all subjects, the most frequently detected species was C.albicans. A statistically significant difference was found between the prevalence of C.albicans (p < 0.001) and C.krusei (p < 0.001) in denture wearers. Subjects with PMMA-based removable prosthetic appliances mostly demonstrated a significant decrease in salivation (p < 0.001), an increase in burning sensations (p < 0.001), and dry mouth (p < 0.001) compared to the subjects who wore partial dentures with Co–Cr metallic frameworks. Red oral lesions were more frequently found among the subjects with partial dentures with Co–Cr metallic frameworks (p < 0.001). Regardless of the material used for the denture, patients must be regularly checked by their dentists in order to prevent the development of oral lesions.