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The olive oil industry generates large quantities of waste pomace which has the potential to be used in a range of applications, including as a source of dietary fibre in the food ingredients sector. This study determined that hexane extracted olive pomace still retained 10.6% soluble dietary fibre (SDF) although the total sugar content of 27% was low. The lower and upper yields from single trials on freeze dried olive pomace (10g) for hexane extracted IDF (insoluble dietary fibre) and SDF were 41–53% and 0.5–2.5%, respectively. These results tentatively indicated that pH and homogenization (high shear mixing) were important factors that affect IDF and SDF yields. Thepilotscale processing of 36kg (wet weight) frozen olive pomace, equivalent to 5.65kg (dry weight), focused on recovery yield of SDF using an alkaline treatment approach combined with wet milling (2h). There was no increase in microbial contamination during the trial. While a relatively high yield of SDF (5.6%) was obtained, themonosaccharidecontent was low, and this fraction did not exhibitgelationproperties which is one of the key indicators for functionality in the food ingredients sector. A lower-than-expected IDF yield (13.6%) was obtained during the pilot trial compared with initial laboratory results (41.3–53.0%). However, this process enriched the fibre content from 40% to more than 70% in the majority of the IDF samples collected during the pilot trial. It was determined that the highest water and oil holding capacities in these samples were 6.9 and 4.1, respectively, which were associated with IDF extracted at pH 4.5. This study revealed that SDF could be recovered from olive pomace and the recovery of IDF could be scaled up, where physical disruption and pH conditions caused apparent changes in the yields, and the water and oil holding capacities of dietary fibre fractions.

Olive mill waste water (OMWW), a by-product created during the processing of olive oil, contains high amounts of polyphenolic compounds. If put to further use, these polyphenolic compounds could be a valuable resource for the speciality chemical industry. In order to achieve this, isolation of the polyphenolic compounds from OMWW is needed. Several techniques for this process already exist, the most widely used of which is adsorption beds. This research describes new ways of collecting polyphenolic compounds by using unmodified iron oxide (Fe3O4) particles and Fe3O4modified with silica gel (Fe3O4@C18), citric acid (Fe3O4@CA), and sodium dodecyl sulphate (Fe3O4@SDS). This approach is superior to adsorption beds since it can be used in a continuous system without clogging, while the nano-sized shapes create a high surface area for adsorption. The results of this study show that, if used in a loop system of several adsorption and desorption cycles, (un)modified Fe3O4has the potential to collect high concentrations of polyphenolic compounds. A combination of different modifications of the Fe3O4particles is also beneficial, as these combinations can be tailored to allow for the removal of specific polyphenolic compounds.

It is important from a techno-economic perspective to understand the challenges associated with developing scalable processes to extract functional proteins from rapeseed cake for applications as functional food ingredients. One of these challenges is to develop a better understanding of the link between upstream mechanical processing of the rapeseed cake and the yield of extracted protein. An initial trial with cold pressed rapeseed cake (CPR) indicated that wet milling had no impact on protein yield. Measurement of protein yields with the Bradford assay indicated that 14% would be recovered. The total protein recovered (5.4% by Kjeldahl analysis) contained aglucosinolateandphytatecontent that was similar to the rapeseed cake at the start of the process. The fate of the protein extraction was followed during a pilot scale trial with hot-pressed rapeseed cake (HPR), using only stirring followed by sieving, revealing an increase in protein yields, as determined by the Bradford assay, from 8.0% at the start to 22.6% at the end of processing. In addition, only 16% of HPR was solubilized. A similar pilot scale trial with HPR using wet milling and stirring followed by decanting also showed an increase in protein concentration from 15.3% at the start to 23.7% at the end of processing. In addition, 38% of HPR was solubilized. This study provides an insight into what the effect of upstream mechanical processing (wet milling) has in releasing protein from rapeseed cake in order to improve process efficiency. This could have important consequences for upscaling and commercial extraction of rapeseed cake protein.

Olive oil production using three-phase decanter systems creates olive oil and two by-products: olive mill wastewater (OMWW) and pomace. These by-products contain the highest share of polyphenolic compounds that are known to be associated with beneficial effects on human health. Therefore, they are an attractive source of phenolic compounds for further industrial use in the cosmetic, pharmaceutical and food industries. The use of these phenolics is limited due to difficulties in recovery, high reactivity, complexity of the OMWW matrix and different physiochemical properties of phenolic compounds. This research, focused on OMWW, was performed in two phases. First, different polyphenol extraction methods were compared to obtain the method that yields the highest polyphenol concentration. Twenty-five phenolic compounds and their isomers were determined. Acidifying OMWW, followed by five minutes of ultrasonication, resulted in the highest measured polyphenol content of 27 mg/L. Second, the collection of polyphenolic compounds from OMWW via adsorption on unmodified iron (II, III) oxide particles was investigated. Although low yields were obtained for removed polyphenolic compounds in one removal cycle, the process has a high capability to be repeated.

Helichrysum italicumis an aromatic plant with promising pharmacological activities. Bioactive compounds found in plants represent an important alternative treatment for weight loss and an infusion ofH. italicumcontains compounds which could have such effect. Our aim was, therefore, to investigate its acute effects on resting energy expenditure (REE) and possible differences in substrate oxidation in a pilot study. A dried, ground plant material ofH. italicumssp.italicumwas infused with hot water and chemically characterized with HPLC-MS analysis. Sensory evaluation of herbal tea was performed. A randomized, crossover, controlled pilot study was then conducted on eleven healthy male subjects. The REE and substrate oxidation were measured by indirect calorimetry at baseline and 30 and 120 min after ingestion of infusion or hot water. The expression of genes involved in lipid metabolism was examined inH. italicuminfusion-treated hepatocytes. Several phenolic classes were identified in the infusion, caffeoylquinic acids were the most abundant, followed by pyrones and flavonols. A single ingestion ofH. italicuminfusion significantly increased REE by 4% and fat oxidation by 12% compared to hot water ingestion. A significant 2-fold up-regulation of β-oxidation-related genes in HepG2 cells, exposed toH. italicuminfusion, was detected. This pilot study suggests thatH. italicuminfusion possesses bioactive substances with potential application in obesity prevention, which could, with additional studies, become an economically interesting novel application of the plant. Clinical trial registration number: NCT04818905

Mediterranean plantHelichrysum italicumrepresents a rich source of versatile bioactive compounds with potential benefits for human health. Despite extensive research on the plant’s active constituents, little attention has yet been paid to characterizing the relationship between its intra-specific genetic diversity and metabolite profile. The study aimed to determine metabolic profile ofH. italicumssp.italicum(HII) and ssp.tyrrhenicum(HIT) cultivated on the experimental plantation in Slovenia and to compare the chemical composition of extracts regarding the solvent extraction process. Extracts were prepared upon conventional extract preparation procedures: maceration with 50% methanol or ethanol and cold or hot water infusion and analyzed using High Performance Liquid Chromatography-Diode Array Detection-Electrospray Ionization-Quadrupole Time-of-Flight-Mass Spectrometry (HPLC-DAD-ESI-QTOF-MS). One hundred compounds were identified in the samples, among them several isomers and derivatives were reported for the first time, while caffeoylquinic acids and pyrones were the most abundant. Semi-quantitative comparison revealed that the extraction procedure had a greater impact on the chemical profile than genetic variability. AllHITextracts showed a higher total phenolic content compared toHII, while the antioxidant potential evaluated by 1,1-diphenyl-2-picrylhydrazil test was not proportionally higher. In addition, hot water extracts proved to be comparably active as alcoholic ones, confirming high commercial potential ofHelichrysum italicumas herbal functional beverages.

Fish products are often sold together with vegetable oil such as olive oil. Lately, the market also offers fish products with extra virgin olive oil...

Jujube (Ziziphus jujuba Mill.) has favourable horticultural properties including adaptation to arid conditions, abiotic and biotic stresses, as well as positive impact on human health. The present study describes the characterization of genetic diversity of the germ­plasm of jujube from the Istrian peninsula, the determination of important chemical compounds, antioxidative properties in relation to antibacterial and antifungal activities of jujube fruit extracts, and the determination of nutritional properties of jujube fruit. The results of the genetic analysis showed that most of the samples from the Istrian peninsula belong to two recently introduced varieties, 'Li' and 'Lang', and the most widespread local variety 'Navadna žižola'. The local variety has smaller fruit than the ‘Li’ and ‘Lang’ varieties, with thick and fleshy mesocarp. Chemical analysis indicated that fruits of the local variety contained a valuable source of dietary fibre ((9.7±0.6) g/100 g) and were rich in minerals such as (in g/100 g dry mass): potassium (829±51), calcium (177±11) and phosphorus (129±19). Aqueous extracts showed slight antibacterial activity, while ethanol extracts had higher mass fractions of phenolic compounds (expressed as gallic acid equivalents (GAE), 5.8-8.7 mg/g) than the aqueous extracts, but did not show antimicrobial activity. Compounds other than phenolic compounds in jujube fruit may be more biologically active. Based on the results of these analyses, the local Istrian jujube variety is a promising candidate for cultivation potential.

In recent years, Slovenian olive growers and producers have faced problems in achieving constant yields and quality of olive oil due to extreme weather conditions, especially because of the more frequent occurrence of drought. A technically accomplished pilot automatic irrigation system for olive trees was established. In 2015, the impact of different irrigation treatments (0, 15, 33, 40 and 100% ETc (crop evapotranspiration)) on olive cultivar 'Istrska belica' were studied to determine the minimum amount of added water that has a positive effect on the yield and quality of the extra virgin olive oil produced. The study results showed that the quantity of applied water equal to 100% ETc had a significant effect on yield of olive fruits. The average fruit mass of the trees grown under full irrigation (100% ETc) was significantly higher than that of those given rain-fed and deficit irrigation treatments (15, 33 and 40% ETc), with no significant differences found between these treatments. The fatty acid contents showed the highest amount of oleic acid (75.61%) at 100% ETc and the lowest amount (74.69%) at 15% ETc, while the highest amount of linoleic acid (6.52%) was determined at 15% ETc and the lowest (5.98%) at 100% ETc. Total biophenol content and biophenol composition was determined. The 40% ETc irrigation system produced the highest amounts of total biophenols, total oleuropein and total ligstroside derivatives (1076, 562 and 388 mg kg-1, respectively), while the lowest results (883, 477 and 299 mg kg-1, respectively) were determined in the control system (0% ETc). The organoleptic assessment of the olive samples showed that the positive sensory attributes of fruity, bitter and pungent were more intensive at 33% ETc and were graded highest. Olive oil from non-irrigated trees (0% ETc) received the lowest sensory score.

Sugars, the primary metabolites of plants, play an important role in the process of olive ripening, as they provide energy for metabolic changes and serve as a source for the biosynthesis of fatty acids. Phenolic compounds, which are secondary metabolites, can be synthesized in plants as a reflection of stressful situations such as tissue damage and UV irradiation. The objective of this study was to assess the impact of different irrigation treatments (0, 15, 33, 40, and 100% crop evapotranspiration (ETc)) on primary metabolites (sugars, sugar alcohols) and secondary metabolites (phenolic compounds). In different phenological periods, the levels of oleuropein, sugars and sugar alcohols were determined in olives and leaf samples in the crop years 2014, 2015, and 2016. The levels of oleuropein in olive fruits and leaves were highest under the irrigation treatment of 40% ETc and decreased noticeably under the irrigation treatment of 100% ETc. The median levels of glucose were higher compared with the mannitol and fructose levels in olive fruits. Total determined sugars in olive fruits during maturation decreased. While significant correlations were found in olive fruits between determined sugars and sugar alcohols and different irrigation treatments, we did not observe any correlation between determined sugars and sugar alcohols in olive leaves and irrigation treatments. Our research confirmed the complexity of the content of oleuropein and sugars or sugar alcohols in the leaves and fruits of olive trees from Slovenian Istra, which depend on various factors, not just drought stress. Weaker and less obvious correlations between the studied compounds and drought stress might be a consequence of unusual rainfall in the studied crop years.

Introduction Neurotoxicity due to acute prenatal exposure to high-dose of mercury (Hg) is well documented. However, the effect of prenatal exposure to low Hg levels on child neurodevelopment and the question about “safety” of fish-eating during pregnancy remain controversial. International comparisons of Hg concentrations in mother-child biological samples and neurodevelopmental scores embedded in birth cohort studies may provide useful evidence to explore this issue. Materials and methods The Mediterranean (Italy, Slovenia, Croatia, and Greece) cohort study included 1308 mother-child pairs enrolled in the Public Health Impact of long-term, low-level, Mixed Element exposure in a susceptible population EU Sixth Framework Programme (PHIME). Maternal hair and venous blood, cord blood and breast milk samples were collected, and total Hg (THg) levels were measured. Demographic and socioeconomic information, lifestyles and nutritional habits were collected through questionnaires at different phases of follow-up. Children at 18 months of age underwent neurodevelopmental testing using the Bayley Scales of Infant and Toddler Development, Third Edition (BSID-III). Multivariate linear and logistic regressions were performed, for each country, to assess the association between THg and BSID-III scores, obtaining adjusted β coefficients and odds ratios (ORs). These values were used to conduct a meta-analysis, to explore possible heterogeneity among countries and to obtain combined estimates of the association between THg exposure and BSID-III scores. Results Median THg (ng/g) was: 704 in maternal hair, 2.4 in maternal blood, 3.6 in cord blood, and 0.6 in breast milk. THg concentrations were highest in Greece and lowest in Slovenia. BSID-III neurodevelopmental scores were higher in Croatia and Slovenia. The meta-analysis of multivariate linear models found an overall positive association between language composite score and receptive communication scaled score and increasing THg in maternal hair (n = 1086; β = 0.55; 95%CI: 0.05–1.05 and n = 1075; β = 0.12; 95%CI: 0.02–0.22, respectively). The meta-analysis of logistic regression models showed that the overall adjusted OR between THg in cord blood and suboptimal gross motor score was borderline significant (n = 882; OR = 1.03; 95%CI: 1.00–1.07). Heterogeneity was found across the four sub-cohorts for language composite score in maternal blood, and for fine motor scaled score in cord blood and breast milk. Language composite score and THg concentrations in maternal venous blood were positively related (n = 58; β = 4.29; CI95% (−0.02, 8.60)) in Croatia and an increase of 1 ng/g of THg in maternal venous blood was associated with a reduced risk for children to fall in the lowest quintile of language score by 31% (n = 58; OR = 0.69; CI 95%: 0.37, 1.01). The comparison of β coefficients obtained by multiple linear regression model showed an inverse association between fine motor score and THg concentrations in cord blood for Croatia (n = 54; β = −0.53; CI 95%: −1.10, 0.04) and Slovenia (n = 225; β = −0.25; CI 95%: −0.49, −0.01). In Slovenia THg level in breast milk was associated with suboptimal fine motor performance (n = 195; OR = 5.25; CI 95%: 1.36, 21.10). Conclusions This study showed an inverse relation between THg levels and developmental motor scores at 18 months, although the evidence was weak and partially internally and externally inconsistent. No evidence of detrimental effects of THg was found for cognitive and language outcomes at these concentrations and age.

The effects of human exposure to mercury (Hg) and its compounds in Europe have been the focus of numerous studies that differed in their design, including recruiting different population groups at different levels of exposure and using different protocols and recruitment strategies. The objective of the present study was to review current studies of Hg exposure in Europe, taking into account the potential routes of Hg exposure, actual Hg exposure levels assessed by different biomarkers, and the effects of Hg to Europeans. All published studies from 2000 onward were reviewed, and exposure and effects studies were compared with known Hg levels in environmental compartments by mapping the various population groups studied and taking into account known sources of Hg. A study of the spatial distribution trends confirmed that the highest exposure levels to Hg, mostly as methylmercury (MeHg), are found in coastal populations, which consume more fish than inland populations. Fewer studies addressed exposure to elemental Hg through inhalation of Hg in air and inorganic Hg in food, particularly in highly contaminated areas. Overall, at the currently low exposure levels of Hg prevalently found in Europe, further studies are needed to confirm the risk to European populations, taking into consideration exposure to various Hg compounds and mixtures of stressors with similar end-points, nutritional status, and a detailed understanding of Hg in fish present in European markets.

This study characterises the genetic variability of local pomegranate (Punica granatum L.) germplasm from the Slovenian and Croatian areas of Istria. The bioactive components and antioxidant and antimicrobial properties of ethanol and water extracts of different parts of pomegranate fruit were also determined, along with their preliminary nutritional characterisation. Twenty-six different genotypes identified with microsatellite analysis indicate the great diversity of pomegranate in Istria. The pomegranate fruit ethanol extracts represent rich sources of phenolic compounds (mean value of the mass fraction in exocarp and mesocarp expressed as gallic acid is 23 and 16 mg/g, respectively). The ethanol extracts of pomegranate exocarp and mesocarp showed the greatest antimicrobial activity against Candida albicans, Candida parapsilosis, Rhodotorula mucilaginosa, Exophiala dermatitidis and Staphylococcus aureus, and the same water extracts against S. aureus and Escherichia coli. To the best of our knowledge, this study represents the first report of the characterisation of pomegranate genetic resources from Istria at different levels, including the molecular, chemical, antimicrobial and nutritional properties., U ovom je radu opisana genetička raznolikost germplazme šipka (Punica granatum L.) s područja slovenskog i hrvatskog dijela Istre. U etanolnim i vodenim ekstraktima različitih dijelova ploda šipka pronađeni su bioaktivni spojevi, te su ispitana njihova antioksidacijska i antimikrobna svojstva i preliminarno određena hranjiva svojstva ekstrakata. Dvadeset i šest različitih genotipova utvrđenih analizom mikrosatelita upućuju na veliku raznolikost šipka u Istri. Etanolni ekstrakti ploda šipka predstavljaju bogat izvor fenolnih spojeva (srednja vrijednost masenog udjela, izraženog kao galna kiselina, bila je 23 mg/g u egzokarpu i 16 mg/L u mezokarpu). Etanolni ekstrakti egzokarpa i mezokarpa šipka imali su snažan antimikrobni učinak na sljedeće vrste: Candida albicans, Candida parapsilosis, Rhodotorula mucilaginosa, Exophiala dermatitidis i Staphylococcus aureus, dok su vodeni ekstrakti egzokarpa i mezokarpa uspješno suzbijali rast bakterija S. aureus i Escherichia coli. Prema našim spoznajama, ovo je prva karakterizacija genetičkih izvora šipka iz Istre, koja obuhvaća molekularna, kemijska, antimikrobna i hranjiva svojstva plodova.

Celtis australis is a deciduous tree commonly known as Mediterranean hackberry or the European nettle tree. The fruit of hackberry are seldom used for nutritional purposes. The nutritional and physicochemical properties of ripe hackberry fruit from Istria (Marasi village near Vrsar, Croatia) were determined, including water, total fiber, protein, vitamin, mineral, and phenolic contents. This analysis demonstrates that the hackberry fruit is a valuable source of dietary fiber, protein, and vitamins, and of pigments such as lutein, β‐carotene, zeaxanthin, and tocopherols. The seasonal differences associated with the different growth stages for the element composition, total phenolic content, and phenolic profile were also determined for hackberry mesocarp and leaves. Water and ethanol extracts were prepared from mesocarp and leaves harvested at different growth stages and their phenolic profiles and antioxidant and antimicrobial activities were investigated. This study demonstrates that water and ethanol extracts of hackberry fruit and leaves collected at different growth stages contain epicatechin, gallic acid, vanillic acid, 3,4‐dihydroxybenzaldehyde, delphinidin‐3,5‐di‐O‐glucoside, cyanidin‐3,5‐di‐O‐glucoside, and pelargonidin‐3,5‐di‐O‐glucoside. They also show some antimicrobial and antifungal activities. Further studies are needed to identify and define the active ingredients of these hackberry leaf ethanol extracts.

Celtis australis is a deciduous tree commonly known as Mediterranean hackberry or the European nettle tree. The fruit of hackberry are seldom used for nutritional purposes. The nutritional and physicochemical properties of ripe hackberry fruit from Istria (Marasi village near Vrsar, Croatia) were determined, including water, total fiber, protein, vitamin, mineral, and phenolic contents. This analysis demonstrates that the hackberry fruit is a valuable source of dietary fiber, protein, and vitamins, and of pigments such as lutein, β-carotene, zeaxanthin, and tocopherols. The seasonal differences associated with the different growth stages for the element composition, total phenolic content, and phenolic profile were also determined for hackberry mesocarp and leaves. Water and ethanol extracts were prepared from mesocarp and leaves harvested at different growth stages and their phenolic profiles and antioxidant and antimicrobial activities were investigated. This study demonstrates that water and ethanol extracts of hackberry fruit and leaves collected at different growth stages contain epicatechin, gallic acid, vanillic acid, 3,4-dihydroxybenzaldehyde, delphinidin-3,5-di-O-glucoside, cyanidin-3,5-di-O-glucoside, and pelargonidin-3,5-di-O-glucoside. They also show some antimicrobial and antifungal activities. Further studies are needed to identify and define the active ingredients of these hackberry leaf ethanol extracts.

The effects of human exposure to mercury (Hg) and its compounds in Europe have been the focus of numerous studies that differed in their design, including recruiting different population groups at different levels of exposure and using different protocols and recruitment strategies. The objective of the present study was to review current studies of Hg exposure in Europe, taking into account the potential routes of Hg exposure, actual Hg exposure levels assessed by different biomarkers, and the effects of Hg to Europeans. All published studies from 2000 onward were reviewed, and exposure and effects studies were compared with known Hg levels in environmental compartments by mapping the various population groups studied and taking into account known sources of Hg. A study of the spatial distribution trends confirmed that the highest exposure levels to Hg, mostly as methylmercury (MeHg), are found in coastal populations, which consume more fish than inland populations. Fewer studies addressed exposure to elemental Hg through inhalation of Hg in air and inorganic Hg in food, particularly in highly contaminated areas. Overall, at the currently low exposure levels of Hg prevalently found in Europe, further studies are needed to confirm the risk to European populations, taking into consideration exposure to various Hg compounds and mixtures of stressors with similar end-points, nutritional status, and a detailed understanding of Hg in fish present in European markets.

The olive variety 'Istrska belica' is well known for its numerous positive properties, such as resistance to low temperature and high oil content. The aim was to determine the variations in the levels of phenolic compounds and sensory properties during storage of 'Istrska belica' virgin olive oil. The profile of the phenolic compounds and sensory properties of 'Istrska belica' olive oil were further compared with those for other varieties, including 'Leccino' and 'Maurino'. The content of phenolic compounds of the olive oils decreased after 1 year and 2 years of storage. After 2 years of storage, the levels of oleuropein and the ligstroside derivates significantly decreased, while the end-stage compounds tyrosol and hydroxytyrosol increased. These data show that after 1 year of storage, the 'Istrska belica' olive oil preserves similar intensities for bitterness and pungency, and similar oleuropein and ligstroside derivates levels. In contrast to the other oils analysed, the intensities of bitterness and pungency of 'Istrska belica' olive oil decreased greatly only after 2 years of storage. Moreover, the phenolic compounds content, and oleuropein and ligstroside derivates levels, and the intensities of bitterness and pungency were the highest in fresh 'Istrska belica' olive oil, compared to the other olive oils analysed. Overall, 'Istrska belica' olive oil has important advantages over olive oil from other varieties that are grown in the Istria region.