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Designing new, environmentally-friendly, and tunable solvents have been dramatically expanding in popularity in order to overcome the flaws of organic solvents from technological, environmental and economic aspects. Natural deep eutectic solvents (NADES), as a new generation of novel alternative solvents, fully meet green and sustainable technologies principles. Though it is sometimes relatively easy to find an appropriate NADES for certain process, this is occasionally ungrateful procedure. Namely, sometimes is difficult to predict which mixtures and in which molar ratios will originate as a NADES and also which solvent properties crucial will be exerted. Therefore, the composition of NADES and their physical, thermal, chemical or biological properties should be characterized on case-by-case scenario. Abundance of possible NADES formulations, especially when possible addition of certain amount of water is taken into account, makes it impossible to prepare and characterize all of them, pointing out the need for development of predictive NADES structure-activity mathematical models. Approaches used so far for screening appropriate/ideal NADES (out of enormous pool of structural possibilities) have been governed by time consuming empirical methods, while systematic investigation in NADES structure-activity relationship, as a backbone for rational design of these solvents, is still lacking. Current literature suggests COSMO-RS, software that can predict thermodynamic properties and phase equilibrium which would help in designing the best possible solvent for a certain application. COSMO-RS can design new solvent structures with tailored properties and thus the trial-and-error method of NADES preparation can be avoided. The aim of this work is to design rationally NADES for the preparation of therapeutic deep eutectic solvents and biocatalytic preparation of chiral drug by combining experimental and modelling tools.

Food processing generates a substantial volume of solid organic by-products. Only small amounts of these by-products are up-graded or recycled and used usually for composting or even discarded in open areas potentially causing environmental problems. Wine production by products, such as grape pomace, are the most abundant worldwide and have been lately often evaluated as a source of biological active compounds due to high content of polyphenolics. In the last few years, the extraction and identification of phenolic compounds from natural sources has become a major area of scientific research. Due to their complex structure, there is no universal extraction method suitable for extraction of all plant phenolic, whereby conventional extraction techniques are usually associated with high organic solvent consumption and long extraction times. One of the organic solvents replacement could be natural deep eutectic solvents (NADES) since NADES can dissolve both polar and non-polar compounds. They may be used for very efficient extraction of natural products from plants, such as polyphenols. Since NADES has high safety profile it is expected that extract obtained by NADES may be directly used in food, pharmaceutical, cosmetical and agrochemical products without the need for expensive downstream purification steps. NADES is designed solvents, so to save time and money, it should be designed rationally for polyphenolics extraction. Currently, COSMO-RS can design new solvent structures with tailored properties and thus the trial-and-error method of NADES preparation can be avoided Therefore, the aim of this stufy is to design efficient NADES for extraction of polyphenols from wine production by-products.

The synergistic use of deep eutectic solvents (DES) and biocatalyst fits logically to the efficient and sustainable production of various commercially interesting products. Namely, biocatalysis ensures catalyzing otherwise difficult transformations in high regio-, chemo- and enantioselective manner at mild and economic conditions, whereby DES can serve as strong green support for modulating/directing reaction route for obtaining the desired product. As the number of structural combinations encompassed by DES is tremendous, it is possible to design an optimal DES for each specific enzymatic reaction system. This important feature of DES allows to (i) enhance enzyme stereoselectivity, activity and stability, as well as reaction rate through enhanced substrate solubility ; (ii) enhance reaction yield (iii) enhance/modify biocatalyst stereo preference ; (iv) contribute to overall greenness of the process (including recycle and reuse). The use of DES for enzyme-catalysed reaction could be considered as “back to the roots”. Namely, when the compounds that constitute the DES are primary metabolites, namely, amino acids, organic acids, sugars or choline derivatives, they can provide cytoplasm-like natural environment for enzymes, meaning that enzymes can transform unnatural substrates in natural environment. When designing efficient biocatalytic process involving DES, independently of reaction type and enzyme used, the following steps should be included: (i) preparation and characterization of DES (ii) screening of DES for optimal enzyme performance (iii) optimization of the biocatalytic process, and (iv) downstream process design with possible scale-up. In this work green lipase-catalysed process for obtaining commercially interesting secondary alcohol (R)-1- phenylethanol of high optical purity in DES as medium was development. Several cholinium chloride-based DES were prepared and screened for optimal enzyme performance (enzyme activity, stability and enantioselectivity), followed by optimization of biocatalytic process (substrate and enyzme loading, water content in DES, reaction temperature). Recovery of the products/DES and DES recycling was successfully performed. After laboratory scale, production of (R)-1- phenylethanol was also effectively performed on a preparative scale (half liter batch reactor). The results of this study confirmed the great potential of DES in biocatalytic medium in terms of improved lipase stability and process sustainability.

Many industrial processes use large quantities of volatile, flammable and toxic organic solvents based on unsustainable resources such as oil, resulting in significant environmental and economic impacts. Over the past decade, natural deep eutectic solvents (NADES) have become promising alternatives to traditional organic solvents from both environmental and technological perspectives. The choice of a solvent in certain process not only depends on its chemical, and physical properties, but also on its environmental impact (e.g. ecotoxicity and biodegradability), sustainability (possibility of recycle and reuse) and process safety (e.g. flammability and volatility). Therefore, interest for potential application of NADES as solvents and/or supporting medium in different processes has increased significantly, due to their unique physical and chemical properties, and low environmental impact. One of the major attractions of making NADES an alternative to conventional organic solvents lies in the fact that the number of structural combinations encompassed by these solvents is tremendous, thus it is possible to design an optimal one for each specific application. Since the number of possible chemical structures of these solvents is vast NADES are very interesting for use in food industry. Potential use of NADES for extraction of biological active compounds, preparation of ready- touse extracts and use of NADES-based extracts for fortification in food industry will be considered herein.

Since NADES consist of simple, inexpensive and naturally occurring compounds with an implied high safety profile, it is assumed that extracts obtained by NADES may be directly used in products for human consumption without the need for expensive downstream purification steps. Considering the potential applications of NADES and NADES-based extracts, it is necessary to further evaluate their biological activity. Human cell lines are already widely used in testing biological activity of compounds present in plants and plant’s products. The application of in vitro cell cultures for this purpose is based on the fact that interaction of the test substance and biological systems occurs preferentially at the cellular level, and has an impact on the whole organism. With such approach a large number of samples, including crude extracts from screening phase, as well as pure fractions obtained during the isolation procedure, could be evaluated. Beyond that, the application of in vitro cell cultures is a good choice for preliminary screen of biological activities, which enables faster and more cost-effective studies compared to in vivo studies. The findings obtained by in vitro tests are valuable and can serve as guidelines for further required in vivo and epidemiological studies of biological activity of plants, its compounds, and products. Based on aforementioned, potential biological activity of plant polyphenolic extracts was evaluated by testing antiproliferative and antioxidant activity. In vitro cytotoxicity of prepared extracts was assessed by antiproliferation assay on two tumour cell lines, whereas for investigation of type of cell death and cell cycle arrest a flow cytometric analysis was applied. Antioxidant activity and possibility of extracts to protect cells from oxidative stress was assessed by ORAC method as well as cell-based methods.

The synthesis of silver nanoparticles (AgNPs) is an important area of research in nanotechnology due to their unusual size- and shape-dependent properties, strong antimicrobial activity and their attractive applications in medicine catalysis optoelectronics, and biotechnology. Various methods can be used for the synthesis of AgNPs such as the sol-gel method, chemical vapour deposition, or thermal decomposition, but these methods are often expensive and employ toxic chemicals. To overcome these drawbacks, biosynthetic approaches using water extracts from different organisms, including fungi, plants and algae, have recently emerged as simple and viable substitutes because they are simple, cheap, and environmentally friendly. Marine macroalgae or seaweeds, such as Fucus virsoides and Cystoseira barbata, are an abundant source of secondary metabolites such as polyphenols, polysaccharides, carotenoids, vitamins, minerals, amino acids, and proteins. Their aqueous extract can act as an efficient metal reducing agent as well as a capping agent to provide excellent stability to the formed nanoparticles. UV-visible spectroscopy is a very useful and reliable technique for the primary characterization of synthesized nanoparticles because the conducting electrons in the outer orbital of AgNPs resonate with specific wavelengths. Reduction of silver nitrate to AgNPs is confirmed with a peak in the range from 335 nm to 560 nm, while the absence of peaks indicates the absence of NP aggregation. In the present study, the water extracts of F. virsoides and C. barbata were used to prepare AgNPs by reducing Ag+ ions from a 1 mM silver nitrate solution to Ag0. Nanoparticle formation was confirmed by UV/Vis spectroscopy, with peaks detected at 380 nm for both algae. The antimicrobial activity of the synthesised nanoparticles against Bacillus subtilis, Pseudomonas aeruginosa Staphylococcus aureus, Salmonella thyphimunum, Enterococcus faecalis, Escherichia coli and Listera monocytogenes was investigated and compared with the antioxidant activity of the seaweed extracts.

Fennel (Foeniculum vulgare Mill.) is a perennial aromatic and medicinal plant belonging to the Apiaceae family. It originates from the Mediterranean area, but it has spread worldwide through naturalization and cultivation. The use of fennel in culinary purposes and traditional medicine is known from ancient times, but nowadays its application has been extended to the pharmaceutical, cosmetic and food industries. Although different parts of fennel (leaves, young shoots, roots and seeds) can be utilized, fennel seeds are mostly exploited due to their abundance of essential oil (up to 5-6%) which is characterized by a strong and pleasant aroma. Fennel essential oil has been recognized to possess antimicrobial, antifungal and antioxidant properties, therefore it is widely used in various areas. The chemical composition of fennel essential oil showed up to 80 volatile compounds, among which trans-anethole, fenchone and estragole are major compounds, being the key compounds of the characteristic fennel flavor. Besides these compounds, the presence of other compounds such as α- and β-pinene, myrcene, α-phellandrene, limonene and p-anisaldehyde have also been reported. The isolation of essential oil can be accomplished by various extraction techniques, such as hydrodistillation (HD) and steam distillation (SD) which are known as conventional ones and still are the most common in use. They are effective for the extraction of essential oils from spices and herbs from which such oils are difficult to isolate and they do not include the use of chemical solvents. On the other hand, their main drawbacks are long duration, difficult regulation of heat and possible loss of thermolabile and light volatiles. Therefore, in order to overcome these deficiencies, advanced extraction techniques have been developed, including microwave-assisted extraction (MAE), enzyme-assisted extraction (EAE), ultrasound-assisted extraction (UAE), subcritical water extraction (SWE) and supercritical fluids extraction (SFE), namely extraction with supercritical carbon dioxide (SC-CO2) which is considered “green” showing the most promising beneficial effects due to being sustainable, environmentally friendly and cost-effective. However, regardless of the selected extraction technique, it is necessary to define optimal process conditions taking into account all process parameters in accordance with the raw material used, and optimize the extraction process with the intention to achieve the maximum yield of essential oil of the best quality and unaltered composition.

Green technologies and their possible applications lately promote the installation of green roofs. In Croatia, there is lack of legislation or regulation for the construction of green roofs. It covers a When planning green roofs, the most attention is paid to the impact of the system on the building ; leakage / drainage, load on the structure, fire protection, etc. Thus living elements such as substrate and vegetation remain on the margins. The rationale for the research was to identify elements that can ensure an integrated approach to planning and implementation of green roof systems. Defined by specific climatic conditions, the possibility of using local materials and soil to create special substrates for green roofs, it is important to determine optimal soil dept level for specific plant categories and the use of indigenous species in the planting plan. The general categorisation of special substrates for green roofs is divided into three main types ; (1) substrate for intensive, (2) simple intensive and (3) extensive green roof systems. They are defined in accordance with the FLL regulations (“Richtlinien für die Ausführung und Pflege von Dachbegrünungen” / “Directives for Green Roof Planning, Implementation and Maintenance”) through: pH value, balanced provision of nutrients, water and air, water holding capacity, drainage and technical suitability. Within these parameters, it is still possible to make a mistake in selecting an adequate system. The microclimatic conditions of the site define the selection of plant species. So, the selected species must be correlated with specific substrate parameters. Plant selection is crucial especially because of long-term maintenance (nutrient content, pest and disease problem, effective drainage, weed reduction) of the green roof system since it is an unrealistic option to expect completely maintenance-free green roof. Also, this research proposes additional expert studies that will propose guidelines adapted to the local climate. It would contribute to the quality and simplicity of installation of green roof systems and give an overview of available options when resources are limited ; economic, technical or plant based.

Both nettle and fennel are valuable plants with high content of bioactive molecules. Beneficial effects of nettle are mainly attributed to polyphenols, while fennel is recognized for its essential oil (EO) content. Although chemically different, both nettle extract and fennel EO are characterized with loss of stability and bioactivity during prolonged storage and processing. Spray drying, as a method for encapsulation of both hydrophilic and lipophilic compounds, can provide stabilization, preservation as well as controlled release and delivery of both nettle bioactives and fennel EO. Therefore, the aim of this study was to evaluate and optimize the spray drying encapsulation of nettle leaves extract and fennel EO using different carrier materials and drying temperatures. Nettle leaves extract was obtained by microwave-assisted extraction and encapsulated using maltodextrin (MD) and β-cyclodextrin (CD) as single carriers or in mixtures with gum arabic (GA) in proportion 1:1 and 3:1, respectively. Carriers were used in sample:carrier ratios 1:1, 1:2 and 1:3, while spray drying was performed at 120, 160 and 200 °C. Fennel EO was spray dried at same temperatures and emulsified with following carrier mixtures, MD:CD, MD:GA and CD:GA in proportions 1:1, 1:3 and 3:1 at fixed EO:carrier ratio of 1:3. The highest yield and encapsulation capacity for nettle leaves extract was achieved with addition of MD as a carrier agent in ratio 1:3 at 120 °C, while optimal encapsulation conditions for fennel EO were mixture of CD:GA in proportion 3:1 at 200 °C which resulted in the highest EO retention in microcapsules and the highest encapsulation efficiency. The differences in obtained encapsulation conditions arise from the differences in chemical properties of encapsulated materials, showing that carrier agents such as CD and GA are suitable for non-polar constituents of EO, while MD is more efficient in encapsulation of polar compounds such as polyphenols. The encapsulation of nettle extract and fennel EO provided preservation and stabilization of respective bioactive compounds and increased their bioavailability in encapsulated form, indicating the great potential of such microcapsules for further application in food industry.

Recycling of waste materials and production of new products with more added values are defined by the principles of the circular economy. Given that the circular economy imply also the continual use of resources and raw materials, this paper deals with the possibility of waste paper use as nano sized filler for cellulose acetete based bio composites. Potential use of multiple times recycled, waste packaging (Kraft) paper and laboratory filter paper as starting materials for nano sized cellulose based filler preparation was determined. As for cellulose acetate polymeric matrix synthesis, Black alder (Alnus glutinosa (L.) Gaertn.) wood and commercial microcrystalline cellulose (Avicel) were used. The optical (UV spectroscopy) and thermal (DSC) properties of prepared bio-nano composites were determined, with supplementary FT-IR analysis of synthesised cellulose acetates. The test results indicated the exceptional importance of the purity of the cellulose base both for the synthesis of nano particles and for the synthesis of the polymeric base of bio- nano composites.

Green infrastructure is term mainly used in recent decade due to Green Infrastructure Strategy realised in 2013th by European commission. Research of evolution of historical towns give us many insights in green infrastructure development (especially urban green infrastructure) because town planning from the past had shown that systematic approach in development of urban open spaces has been conducted many centuries ago. Present situation indicates many formal recommendations of its implementation emphasised with Green Plan from 2020th year. This research shows what kind of changes can be expected in urban planning in future and in which direction our towns are going to be developed. Sustainability, as a key concept of the future urban spaces, needs to envision many changes in urban planning and in policies connected to it. This changes in traditional, less flexible, spatial and urban planning processes needs to accommodate new way of thinking, conceptualising with public participation in development of plans. In these transformational processes concerns in lack of environmental empathy of society in hole (including different stakeholders – planners, local authorities, citizens etc.) is going to be crucial in Croatia. There are highly visible needs for education, transparency and accessibility of different stakeholders. In these processes knowledge based planning, mainly in bioecology and urban morphology premises is going to be crucial. Experiences in landscape studies of different towns are giving us insight of diverse problematic agenda which needs to be addressed in this transformation.

Urban forest ecosystems have their own specific climate or microclimate in relation to other parts of cities. The aim of the research was to determine and analyse the microclimate of urban forest ecosystems in the city of Zagreb. Microclimatic conditions were measured at five different locations in the urban forests of the city of Zagreb. The locations where the measurements were performed were the park of the Faculty of Forestry, University of Zagreb, Forest Park Maksimir, meadow in Forest Park Maksimir, second Maksimir Lake, and avenue of wild chestnuts (Aesculus hippocastanum L.) in Maksimirska Street. At 7 h, 14 h and 21 h for ten consecutive days in July, air temperature (° C), relative air humidity (%), wind speed (m / s), soil temperature (° C) and soil volumetric water content were measured. The comfort index is calculated based on the values of air temperature, relative air humidity and wind speed. During the ten days of July, the mean values of air temperature (24.0 ° C), soil temperature (24.9 ° C) and wind speed (1.0 m/s) were the highest in the tree line in Maksimirska Street. The mean value of relative air humidity was highest in the Forest Park Maksimir, and the soil volumetric water content along the lake in the Park Forest Maksimir. In contrast, mean air and soil temperature values were lowest in the Forest Park Maksimir. The mean values of relative air humidity were the lowest in the tree line in Maksimirska Street, and the soil volumetric water content in the Forest Park Maksimir. According to the comfort index for the human body, the park, forest park and lake were "comfortable", while the meadows and tree-lined avenues in Maksimirska Street were "fresh" for the human body. Wind speeds were higher in more open locations such as park, meadow and tree-lined avenue in Maksimirska Street, while they were smaller in locations with a complete set of canopies, such as in the Forest Park Maksimir and by the lake. The significance of these measurements is in determining the contribution of forest vegetation to the modification of the microclimate of urban ecosystems, which contributes to a more pleasant stay and recreation of residents in these locations.

Climate change and the growing world population are affecting every human activity, especially agricultural production. The main goal is to produce adequate yields of high-quality plant material rich in nutritive value. One of the possible solution to solve the above-mentioned problems is the introduction of wild species into agricultural production. Wild species such as stinging nettle (Urtica dioica L.) and fennel (Foeniculum vulgare Mill.) are plant species that have a strong biological potential and nutritive value whether wild or grown under controlled conditions (cultivated). The aim of the present research was to test the influence of different fertilization treatments on the quality of wild nettle and fennel. A split-plot experiment was set with four fertilization treatments as factors – control (no fertilization), mineral fertilizer, organic fertilizer and a combination of mineral and organic fertilizers, set on both stinging nettle and fennel. Fennel experiment results showed a significant increase in the vegetation index under mineral fertilizer application, compared to other treatments. Both mineral and combination (mineral and organic fertilizer) resulted in higher yield. Stinging nettle experiment results showed a significant increase in vegetation index under mineral, organic and combination treatments application, compared to control treatment. Furthermore, the same results were obtained regarding stinging nettle yield. It can be concluded that the introduction of wild species such as stinging nettle and fennel into agricultural production can be successfully achieved and fertilizer application showed a positive effect on both vegetation index and yield.

materials for different biotechnological processes [1, 2]. Pre-treatment of such materials is necessary due to their complex structure. With desirable products formed during pretreatment as glucose and xylose for example [1] a variety of undesirable by products are formed as organic acids and aromatic compounds (eg. furfural, formic acid, hidroxymetil furfural, levulinic acid, acetic acid)[3]. Those compounds have inhibitory effect on selected microorganism and on bioprocess efficiency [1, 4]. Origin of inhibitors can be categorized in three notable groups. First group are inhibitors present in lignocellulosic structure. Compounds as aldehydes and phenols are created during depolymerization of lignin. Acids as acetic, formic, galacturonic are formed during degradation of hemicellulose [4]. Second group of inhibitors are compounds that are formed during degradation of lignocellulosic materials. In the structure of lignin, generally they are present as non-inhibitors (pentose and hexose sugars). Under high temperature and acidic conditions, pentose sugars are degraded to furfural (furan 2- carbaldehyde) and hexose sugars to 5- hydroxymethyl furfural. Also 5- hydroxymethyl furfural can be degraded to levulinic acid and formic acid [3, 4, 5]. Third group of inhibitors are compounds present on lignocellulosic materials and are not integral part of materials itself. Most common are nitrogen and sulphur chemical compounds, heavy metals and different pesticides or other chemicals used in agriculture [3, 4, 5]. Furfural and hydroxymethyl furfural are two most important inhibitors that are created during lignocellulosic pre-treatment. Influence of furans (furfural and hydroximethylfurfural) on the biomass growth and activity can be used as an inhibition indicator for other compounds present in hydrolysates too [6, 7]. Set of experiments were carried out to investigate the inhibitory effect of furfural on the growth and ethanol production with wild strain Saccharomyces cerevisiae. Selected microorganism is potential ethanol producer from hydrolysates obtained with weak acid pre- treatment of lignocellulosic materials as wheat straw.

Lactic acid is mainly produced by lactic acid bacteria (LAB ; Lactobacillus sp., Streptococcus sp., Pediococcus sp.) and molds (e. g. Rhizopus oryzae, Rhizopus arrhizus)[1]. This bulk chemical can be produced from first and second generation raw materials [2]. First generation raw materials are feedstocks that contain sugar and starch and are widely used in industrial-scale lactic acid production. Second generation raw materials are ligncellulosic raw materials that have to be pretreated prior to fermentation [3]. During pretreatment variety of toxic compounds like furfural, hydroxymethylfurfural, levulinic and acetic acid and different lignin derivatives are beeing formed [4]. Obtained hydrolysates are usually treated by different expensive and long-lasting methods that can be used for detoxification procedures. Therefore, tolerance of working microorganism to above mentioned toxic compounds could be an asset in lactic acid production process [5]. Furans can be used as good predictor of the inhibition effect of other toxic substances present in lignocellulosic hydrolysate [2] and furfural was identified as main inhibiting compound obtained after dilute acid pretratment of lignocellulose [6]. Since there is limited amount of data available on toxic effect of inhibitors formed during pretreatment of lignocellulose [6], in this work the influence of different concentrations of furfural (from 100 to 5000 mg L-1) on growth and activity of lactic acid bacterium Lactobacillus amylovorus DSM 20531T in De Man-Rogosa-Sharpe (MRS) medium was investigated.

In this study, activated carbon derived from Amorpha fruticosa L. was evaluated as a potential adsorbent for removal of methylene blue dyes (C.I. 52015). The adsorption process with respect to operating parameters (contact time, pH, adsorbent dose, temperature, concentration) was investigated to evaluate the adsorption characteristics. Research of equilibrium, kinetics and thermodynamic studies was performed. The results indicated that the maximum amount of adsorbed dye (99, 78%) was reached at pH 5, 5 and at 20 min. This study suggests a potential use of Amorpha fruticosa L. for the removal of cationic dye from aquatic media.

The essential oils have been in use for many years in industries, especially in the pharmaceutical, cosmetic and sanitary industry. Recently, many essential oils are used in culinary and food industry as spices and flavouring agents. Aluminium alloys with magnesium are one of the most important construction materials, especially in shipbuilding, but also for use in the chemical and food industry. Despite the good properties of Al-Mg alloys, they are not perfect materials for engineering applications in all environments, since they suffer from corrosion caused by chemical interactions with their surroundings. For the selection of aluminium and its alloys for the needs of the food industry, their good corrosion resistance to most food products is particularly important. High corrosively environment for aluminium and aluminium alloys in food industry are foodstuffs with pH 3 – 5, such as fruit juices, canned fruits, sauces as well as vegetables and fish pickled in brines with 1 –3 % salt. For this reason, NaCl and acetic acid needs to be taken into corrosion consideration, due to its wide usage in food industry. This study was carried out on AA5754 alloy (aluminium alloy with 3% of magnesium) using gravimetric and potentiodynamic polarization methods, while the alloy surfaces were examined by scanning electron microscopy (SEM). Corrosion study of AA5754 alloy in 3 % NaCl, 1 % CH3COOH and their mixture with addition of 20 ppm laurel, basil and dill essential oils as three very commonly used herbs to improve food flavorings, was conducted in temperature range 25°C to 80°C. Results show that the best inhibitory action on Al-Mg alloy corrosion have basil oil at all investigated temperatures, especially in mixture of 3% NaCl and 1% CH3COOH solutions.

Sugar and starch containing raw materials are widely used as food and feed thus, second generation raw materials like waste lignocellulosic biomass present sustainable alternative for production of different biochemicals, e.g. lactic acid. In this work, alkali pre-treatment of wheat straw with 2 % w/w sodium hydroxide solution in high-pressure reactor at different temperatures (from 120°C to 210°C) and residence times (from 1 to 20 minutes) was performed. After alkali pre-treatment of wheat straw, solid and liquid phase was obtained and composition of the two phases was determined. Solid phase (wheat straw cellulose) with high glucan content was used to study the fermentation capabilities of selected strain in two bioprocess types: Separate Hydrolysis and Fermentation (SHF) and Simultaneous Cellulose Hydrolysis and Fermentation (SCHF). Cellulose hydrolysis was performed with commercially obtained cellulases and β-glucosidases isolated from mold Aspergillus niger. In SHF bioprocess, after enzymatic hydrolysis of wheat straw cellulose, obtained hydrolysates were used for lactic acid production by lactic acid bacterium Lactobacillus amylovorus DSM 20531T. While, in SCHF bioprocess, enzymatic hydrolysis of wheat straw cellulose to simpler carbohydrates and fermentation by L. amylovorus DSM 20531T was performed simultaneously. Process efficiency parameters, like final lactic acid concentration, lactic acid yield and bioprocess productivity, in SCHF bioprocess were approximately 60 % higher than in SHF bioprocess.

The volatile compounds from virgin olive oil (VOO) are chemical species responsible for its specific flavour. This group of compounds consists mainly of C5 and C6 volatiles that are biogenerated from polyunsaturated fatty acids via lipoxygenase pathway. In this paper the composition of volatile compounds, total phenolic content and sensory profile of monovarietal VOOs from four Dalmatian most common cultivars: Oblica, Lastovka, Levantinka and Krvavica were studied. Fruits from each variety were handpicked at the same ripening stage and processed separately in oil under the same conditions using laboratory oil mill Abencor mc2. All analyzed VOOs were within limits established for extra virgin olive oil category based on chemical parameters. Total phenolic content (TP) of obtained oils were determined spectrophotometrically using modified Gutfinger method and expressed as caffeic acid. The highest TP value was detected in Oblica and Levantinka oils (482.32 mg/kg and 398.85 mg/kg, respectively). Volatile composition of analyzed monovarietal olive oils was determined using headspace solid-phase microextraction with gas chromatography/mass spectrometry (HS-SPME/GC-MS). The most prevalent volatile compound was C6 aldehyde E- 2-hexenal, that contributes to green, fruity, bitter and astringent sensory characteristic of olive oil. The highest value for E-2-hexenal was detected in Levantinka oils (75.89% of total peak area), followed by Lastovka (55.27%) and Oblica (54.86%). In the same time, Lastovka oils had the highest ammount of hexanal (25.72%), C6 aldehide associated with sensory characteristics of green fruitiness, apple and grass. Quantitative descriptive sensory analysis of studied olive oil samples were carried out by a sensory panel using IOC methodology. The sensory profile of Lastovka oil was characterized by medium intensity of olive fruity, green leaves, apple and almonds, strong bitterness and medium pungency. Krvavica oil sensory profile showed the mildest intensity of bitter and pungent of among all analysed oils. Mild to medium fruitiness with grass and ripe apple notes are olfactory characteristics of Krvavica oils.

Adsorption is the method most often used for the removal of dyes from industrial effluents. However, due to the high-cost of conventional adsorbents, a wide range of lignocellulosic waste materials are being extensively investigated as a possible novel adsorbents. Many of them proved to be very effective. Dye-loaded lignocellulosic material cannot be directly discarded to the environment, so one of the alternatives is to ferment it using fungi that are capable of degrading the dye. For the purpose of future analyses of fermented dye-loaded lignocellulosic material, i.e. the quantification of the dye degradation extent, a series of organic solvents were used to extract the methylene blue (MB) adsorbed onto brewers’ spent grain (BSG). Both unfermented and fermented (solid-state fermentation using T. versicolor) dye-adsorbed BSG samples were tested. The individual solvents used in this study included nonpolar slovents (benzene, toluene, chloroform, dichloromethane and xylene), polar aprotic solvents (tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide), and polar protic solvents (ethanol, methanol, amyl alcohol and water). Furthermore, a two-component solvent system (methanol : water 1 : 1) and three-component solvent system (methanol : chloroform : water 1 : 1 : 1) were also used. To stress the importance of the principles of green chemistry, ultrasonic-assisted extraction of MB from BSG with deep eutectic solvents (DES), choline chloride/hydrogen bond donor (ChCl/HBD) was also explored. The results showed that the highest desorption of MB from BSG was achieved using polar protic solvents, out of which methanol was the best. The lowest desorption was achieved using non-polar solvents. The results of the MB extraction from BSG using deep eutectic solvents indicated that some of the tested eutectic solvents could be used as an “green” alternative to organic solvents. However, they were far less efficient than the polar protic solvents.

Dandelion ( Taraxacum officinale) leaves and roots are good sources of health-promoting phytochemicals. Generally, applications of modern green extraction techniques, such as High Hydrostatic Pressure Assisted Extraction (HPAE) and Pressurized Liquid Extraction (PLE) showed significant advantages over conventional extractions in terms of less solvent and energy consumption, shorter extraction time, higher yields, etc. Thus, it could be useful for the effective extraction of various bioactive compounds from plant materials. In line with this, the aim of this study was to investigate the use of HPAE and PLE for the recovery of polyphenols from dandelion leaves and roots. Studied parameters for HPAE were: ethanol concentrations (50 and 70%), high pressures (300, and 500 MPa), times (5 and 15 min), and temperatures (22 and 60°C) PLE was conducted at constant static time (5 min) and pressure (10.34 MPa) by varying: ethanol concentrations (50 and 70%), temperatures (60, 80 and 100 °C), and number of extraction cycles (1, 2 and 3). Optimal HPAE parameters for the polyphenols recovery of dandelion leaves and roots was at 5 min, 60°C, and 500 MPa, with 70% ethanol (v/v) that was the most effective solvent for the leaves ; and 50% ethanol (v/v) for the dandelion root extract. Dandelion leaves extract was richer with polyphenols than the root extract (44.99 vs.11.46 mg GAE/g). With respect to PLE, 50% ethanol (v/v) was the most appropriate extraction solvent from both plant matrices. Moreover, optimal PLE conditions with the highest recovery (93.94 mg GAE/g) for the leaves were 60°C with 2 extraction cycles, while for the roots highest yields (5199 mg GAE/g) were achieved under higher temperature and longer extraction time (80°C and 3 extraction cycles). In conclusion, both extraction techniques were suitable for the efficient green recovery of polyphenols from dandelion leaves and roots. However, according to obtained results PLE performance has proven to be more suitable than the HPAE. Therefore, it should be recommended for the fast and effective polyphenols recovery from both, leaves and roots.

Brewers’ spent grain (BSG), being the food industry lignocellulosic waste, was tested for its ability to adsorb azo dye Congo red (CR) from aqueous solutions. Effects of the initial dye concentration (30, 50, 100 and 150 mg L-1), contact time, adsorbent concentration (5, 10, and 15 g L-1), and pH (7, 8, 9 and 10) on dye removal were investigated. Dye removal was rapid within the first 60 min of the contact time. The percentage of dye removal increased with the increase in the adsorbent concentration and decrease in the initial dye concentration. The increase in pH caused reduction of the percentage removal. The percentage of CR adsorptive removal after 240 min contact time was over 85% for all runs. Dye-adsorbed biosorbent was further used as a substrate for solid-state fermentation (SSF) by the white rot fungus Trametes versicolor CCBAS AG613. Prior to solid-state fermentation experiment four white rot fungi strains, Phanerochaete chrysosporium CCBAS 570, T. versicolor CCBAS AG613, T. versicolor TV6 and Ceriporiopsis subvermispora, were screened for their ability to decolourise CR during cultivation (9 days at 27 ⁰C) on agar plates containing dye at final concentrations of 50, 100 and 150 mg L-1. Radial growth and the zone of colour change on agar plates were measured (at two perpendicular directions) every three days. Fungus T. versicolor CCBAS AG613 showed the highest decolourisation index (decolourization diameter/mycelial diameter) and was chosen for further SSF experiment. SSF of dye-adsorbed BSG (250 g BSG soaked in 350 mL of 150 mgL-1 dye solution) was carried out in a horizontal cylindrical glass flask at an ambient temperature. Temperature and weight were monitored on-line, while moisture, protein content, and colour of samples were measured after 7, 14 and 21 days of fermentation. Continuous protein content increase and weight loss were observed as a result of fungal growth and material biodegradation. Complete decolourisation of samples was not observed. However, colour determination showed the continuous increase of total colour change and colour intensity change of fermented samples, compared to their abiotic controls.

1) microwave extraction of Dalmatian sage (Salvia officinalis L.) extracts that aimed to evaluate the influence of solvents (30% ethanol/water, v/v ; 30% acetone/water, v/v ; water), time (3, 5, 7, 9, 10 min), temperature (30, 50, 60, 80 °C), and acidity (addition of 10 % HCl) on polyphenolic recovery. Total polyphenols were best extracted with 30% acetone/without HCl/T=80°C for 10 minutes. The main polyphenol, rosmarinic acid was best extracted with 30% ethanol/with HCl at T=80°C for 10 minutes ; 2) cold atmospheric gas phase plasma extraction of anthocyanins from pomegranate (Punica granatum L.) juice. Plasma treatment covered different operating settings: (i) time (3, 5, 7 min), (ii) juice volume (3, 4, 5 cm3), and (iii) plasma gas flow (0.75, 1, 1.25 dm3/min). Anthocyanins were best stable at: 3 min of treatment time, 5 cm3 sample volume, and 0.75dm3/min gas flow. Anthocyanin content was higher for 21-35% in plasma treated samples vs. fresh juices. In summary, it was shown that plasma treatment had positive influences on anthocyanins stability in cloudy pomegranate juice ; 3) cold atmospheric gas phase plasma of chokeberry juice (Aronia melancarpa) extracts that aimed to evaluate influence of different treatment times and juice volumes under constant gas flow (0.75 dm3min-1) on polyphenolic content. All samples were compared to the fresh and pasteurized chokeberry juice (80°C/2min). Plasma treatment better preserved hydroxycinnamic acids, but showed loss of anthocyanins. Pasteurisation negatively affected hydroxycinnamic acids, while flavonols and anthocyanins were well preserved.

The study presents an analysis of precipitation, runoff and sediment transport on micro-scale plots, on bare ground and on area covered by grass and Spanish broom (Spartium junceum L.), in the Suvava catchment (central Dalmatia, south Croatia). The hydrological experimental plots were set parallel to the inclination of 16°, on marl lime stone and shallow and very skeletal soil (calci-mollic cambisol) and are equipped with rain gauge. At the plots the runoff and suspended sediment were monitored after every rainy day. The monthly runoff was highly variable, ranging between 0.10 to 4.40 mm m-2 on the plot covered by Spanish broom ; between 0.30 to 9.86 mm m-2 on the plots with grass cover and between 1.10 to 21.70 mm m-2 on the bare ground. The monthly sediment transport also was highly variable. It was ranging between 0.04 to 0.71 kg ha-1 on the plots covered by Spanish broom ; grass cover between 0.06 to 11.68 kg ha-1 and on bare ground between 0.4 to 558.1 kg ha-1. Average annual runoff on bare ground was bigger for 10 times and on the plot covered by grass 4 times than on plot covered by Spanish broom. Annual coefficients of runoff on the investigated types of vegetation cover were small and did not exceed 10 % of total annual rainfall. Sediment transport on bare ground plot was bigger 690 times and under grass cover 13 times than on plot covered by Spanish broom on the same inclination. Research results showed that vegetation cover on experimental plots has big impact on the runoff and sediment transport. Runoff and sediment transport are in strong linear dependence on rainfall quantity, duration and intensity.

In recent years the consumers have been showing an increased interest in chestnut fruits because of their nutritional qualities and potential beneficial health effects. Since the information about nutritional composition of the fruits from the natural populations is still limited the aim of this stud was to analyze their mineral content. Nuts were sampled in five natural populations, from the central area of sweet chestnut distribution range in Croatia. Each population was represented with five trees. Macro and micro elements (K, Ca, Mg, Na, Fe, Zn) and heavy metals (Pb, Cd, Hg, As) were determined by atomic absorption spectrophotometry using a Varian SpectrAA 220 device. Univariate and multivariate statistical techniques were used to evaluate the differences among populations. Chestnuts of all five populations contained K, Ca, Mg, Na, Fe, Zn in average 613.96, 35.17, 29.99, 3.69, 1.37 and 0.70 respectively. The obtained results for heavy metals content provided evidence that the concentrations were lower than the toxic law limits. An analysis of variance showed significant difference between populations regarding each mineral. The study also confirmed a trend of population variations according to the eco-geogrphic principle.

Cryphonectria parasitica (Murr.) Barr is an introduced phytopathogenic fungus and causal agent of chestnut blight, a devastating disease of chestnut trees. Since its introduction into Europe in the first half of the 20th century it has caused serious damage in the stands of European chestnut (Castanea sativa Mill.). After initial disease outbreak chestnut populations started to recover due to the natural biocontrol of C. parasitica with cointroduced dsRNA virus Cryphonectria hypovirus 1 (CHV1). CHV1 reduces the virulence of C. parasitica, a phenomenon called hypovirulence, thus enabling the recovery of chestnuts. Based on the genetic variability of European populations five subtypes of CHV1 have been defined, differing in their virulence towards the fungal host. In several European countries chestnut blight biocontrol by application of selected hypoviulent C. parasitica strains on active cankers is implemented. The success of biocontrol is limited by high diversity of fungal vegetative (vc) types which impedes the spread of the virus through C. parasitica populations. Thus the best practice approach in the biocontrol has to take into account specificities of local C. parasitica and CHV1 populations. The aim of the work was to investigate the effect of Croatian virus strains on different Croatian C. parasitica isolates in order to find optimal virus candidate for use in biocontrol treatments of chestnut blight in Croatia. Virus isolates were characterised as “strong” or “weak” based on their impact on fungal growth on chestnut stems. Then they were transferred into three fungal isolates of the same vc type by hyphal anastomosis. The effect of the virus on fungal radial growth in vitro and on the activity of fungal stress response enzymes catalase, glutathione S-transferase and superoxide dismutase was measured. The presence of CHV-1 in the C. parasitica resulted in reduced fungal growth in most virus-fungus combinations. The activities of fungal stress enzymes were affected by the presence of CHV-1, but the direction and intensity of the effects depended on virus and fungus isolate and their combination. Therefore, the “strength” of the virus is not solely determined by its subtype but is very strongly affected by both host and pathogen genotypes. This further confirms that possible biocontrol efforts have to take into account local specificities of C. parasitica and CHV1 populations.

Forest fuel is by nature a very diverse source of energy. Most of its properties are given and limit the possibility of upgrading the fuel in the forest production process. Because of that, the ones that can be manipulated during the transformation from raw material source (tree biomass) to useful and easy to handle forest fuel form (wood chips) are of great importance. This research was conducted in order to determine the impact of raw material preparation by different harvesting systems on the quality of produced wood chips in respect to particle size distribution and ash content. The research material was procured in a spruce plantation thinning operation. It consisted of pulp wood and whole trees harvested with partially mechanized harvesting systems (chain- saw and skidder) and of pulp wood, stem wood and tops harvested with mechanized harvesting systems (harvester and forwarder). The roadside chipping of the unseasoned raw material was done by a drum chipper. Sampling of produced wood chips was done on a total of 21 chipping cycles (truck loads) and laboratory analyses were performed in accordance with the HRN EN standards for Solid biofuels. Results of laboratory analyses were statistically processed. Analysis of variance on the quality parameters of different raw material types was performed and correlation between the quality parameters was tested. The median value of particle size distribution showed statistically significant difference in between the raw material groups. A negative correlation was found between the median value of particle size distribution and the ash content. Results of this research indicate a possibility to manipulate the natural ash content of the forest fuel (a given value that varies mainly with the tree part used as raw material) and the polluting ash content (a consequence of (miss)handling in the procurement chain) to minimize the fuel quality deterioration. Results obtained in this research can be regarded as an indicator of necessity to expand the research-based knowledge to various stand and operational conditions, as a crucial tool of the forest fuels quality control in order to produce the forest fuel of the best possible quality even from the lower quality raw material and more important to avoid that from the high-quality raw materials a forest fuel of poor quality is produced.

Water is the essence of life, as well as an important raw material in many industrial processes. Due to rapid economic development and often irresponsible behaviour of mankind towards nature, in many cases the resources of clean water have been endangered. Therefore, water management is a significant part of human prospect and welfare. From the environmental point of view, the possibility of water savings in every field of human activity is very desirable. Since the industrial sector demands huge amounts of water, it is extremely important that industry adopts a concept of various water savings possibilities. This primarily applies to those industries, where health properties of water are not the most important issue, such as the case in manufacturing of bitumen based products, construction works, concrete preparation, the use of water as cooling fluid in power-plants… In this work the study of wastewater reuse in manufacturing of bitumen based products have been studied in one production plant in the North West region of Croatia. A special focus has been put on water reuse and findings of possible solutions regarding minimisation of the amount of fresh water usage, as well as the costs of water drainage in the production cycle.

Water is the essence of life, as well as an important raw material in many industrial processes. Due to rapid economic development and often irresponsible behaviour of mankind towards nature, in many cases the resources of clean water have been endangered. Therefore, water management is a significant part of human prospect and welfare. From the environmental point of view, the possibility of water savings in every field of human activity is very desirable. Since the industrial sector demands huge amounts of water, it is extremely important that industry adopts a concept of various water savings possibilities. This primarily applies to those industries, where health properties of water are not the most important issue, such as the case in manufacturing of bitumen based products, construction works, concrete preparation, the use of water as cooling fluid in power-plants… In this work the study of wastewater reuse in manufacturing of bitumen based products have been studied in one production plant in the North West region of Croatia. A special focus has been put on water reuse and findings of possible solutions regarding minimisation of the amount of fresh water usage, as well as the costs of water drainage in the production cycle.

Within ‘green economy’ approach, growing area of research in the development of green technologies is devoted to designing new, more environmentally friendly solvents. As a promising alternative to traditional organic solvents deep eutectic solvents (DESs) have been dramatically expanding in popularity last few years as a new generation of designer solvents with possible applications in various industrial fields. Herein, we will present recent progress of our multidisciplinary studies on DESs application in biotechnology. Within this study a number of DESs based on renewable sources (e.g. choline chloride, sugar, amino acids and carboxylic acid) were prepared, and screened for their applicability as green solvents in extraction of phenolic compounds from grape skin and lipasecatalyzed synthesis of short chain ester. In order to fully define DESs as environmentally benign, and thus the processes involving them, toxicity profiles of these solvents were also studied. The data obtained would serve to fill the existing gaps in the knowledge regarding DESs application in the extraction of various natural products from plant material as well as in biocatalytic processes. Data present here are our proof-of-concept for possible implementation of DESs into truly green industrial processes.

Mycological and plant characteristics of Austrian pine (Pinus nigra Arnold) stands in Istria, Croatia were surveyed. Mycocoenological description was based on fruit body and species number. Fungi samples were collected on three different permanent research plots of 400 m2 each, for three consecutive years (2011/2012/2013) from week 36 to week 50 every fortnight. Plots showed distinct differences in fungal species and fruit body number. High variation of fruit bodies for the single year was observed. Stands characteristics were also observed in order to define potential natural vegetation status of pine cultures. Phytocoenological survey was performed during April and June of 2014. at all plots. Three soil types recorder were Eutric Cambisol on flysch bedrock, Terra Rossa and Black soil on limestone (Calcomelanosol). Floristic composition of pine stands differed among the plots showing mixtures between eumediterranean/submediterranean and submediterranean/ mountain vegetation regions. Together with plants, different fungi species were found as good indicators of Austrian pine habitat characteristics. Long term survey is needed to better understand the ecology and role of certain fungal species that characterize different Austrian pine habitats.

For successful afforestation in Mediterranean karst areas several important criteria must be met, especially adequate tree species and high-quality seedlings. With that in mind, morphological features of one-year old Aleppo pine (Pinus halepensis Mill.) seedlings were analyzed. Seedlings were grown in different types of containers to establish the effect of container type on development (quality) of above- and belowground parts of the seedlings. Four types of containers were used: MP53/12, MP33/18, T7/24 and T8/24. Studied seedling variables (morphological features) include: seedling height, root collar diameter, biomass of the aboveground part of the seedlings and total seedling biomass. Two morphological indexes were calculated from measured variables: S/R ratio and DQI. Morphological dimensions of root systems were scanned and measured with software WinRhizo, namely: total root length, number of root tips, surface area and root volume. Morphological features with regard to container type were evaluated with analysis of variance (ANOVA) and multiple Turkey's post hoc test. Morphological features of seedlings derived from the samples of 20 seedlings per container type, almost all show significantly higher values in larger containers (T7/24 and T8/24) compared to smaller containers (MP53/12 and MP33/18). Root analyses also show that roots of seedlings from smallest containers (MP53/12) are deformed to a high degree. In larger containers (MP33/18) root deformation is largely reduced. On the other hand, bind of roots into growing medium in seedlings grown in large containers is insufficient, although the roots are not deformed. Seedlings of Aleppo (Pinus halepensis Mill.) grown in container MP53/12 have developed highly deformed root systems because of inadequate dimensions of this container and its technological absoluteness. Therefore is recommended to gradually abandon the use of MP53/12 and MP33/18 containers from nursery production. Further research is needed to eventually define new, modern solutions adapted to production of high-quality seedlings for afforestation of Mediterranean karst area.