Hydrodistillation and Microwave Extraction of Volatile Compounds: Comparing Data for Twenty-One Veronica Species from Different Habitats.

Free volatile compounds were isolated from 21 Croatian Veronica species studied by hydrodistillation (HD) and microwave extraction (ME) and analyzed by gas chromatography coupled with mass spectrometry. Principal Component Analysis (PCA) distinguished some clusters based on the relative proportion of major compounds, such as hexadecanoic acid, hexahydrofarnesyl acetone, phytol, E-caryophyllene, and caryophyllene oxide, which were identified in all species studied by both isolation methods. In addition to these compounds, germacrene D, δ-selinene, and eicosane were also identified in five samples from dry habitats isolated using ME. Allo-aromadendrene and ß-ionone are particularly abundant in five species from wet habitats isolated by both methods. The peculiarities of Veronica species from moderate habitats isolated with HD are benzene acetaldehyde, n-nonanal, and the identification of significant compounds from the hydrocarbon class, while the peculiarity of ME is (E)-ß-damascenone. In this article, we present new results on the phytochemical characterization of Veronica species from different habitats. The biological potential of these compounds should be further investigated for a better understanding and utilization of the specialized plant metabolites.

Physicochemical and aromatic characterization of carob macerates produced by different maceration conditions.

Carob liqueur is an alcoholic drink (minimum 15% v/v of ethanol and 100 g/L of sugar) typical for the Mediterranean countries. In the current work, carob macerate produced by maceration of carob pods in hydroalcoholic base at different maceration conditions was characterized for the first time based on its aroma compounds/profile, physicochemical parameters, and chromatic characteristics. The results confirm the migration process of bioactive compounds, aroma compounds, and sugars flowing from the carob pod to the hydroalcoholic base. Changes in ethanol concentration modify the physical properties of the solvent and influence the phenolic and aroma compounds extraction, color, and acidity of the obtained samples. The higher content of phenolic compounds was determinate in the samples obtained in the darkness. The amounts of phenols were in the range of some red fruit liqueurs or walnut liqueurs, and sugars (mostly sucrose) ranging between 96 and 107 g/L. Twenty-six (out of total 94) aroma compounds were detected in all samples, of which 17 esters, 3 alcohols, 4 ketones, and 2 acids. Low molecular weight ethyl esters, ethyl hexanoate, ethyl 2-methyl propanoate, ethyl octanoate, ethyl benzoate, ethyl butanoate, and ethyl cinnamate, were the most abundant. Carob pod maceration in 50% v/v hydroalcoholic base (1:5 solid to liquid ratio) in darkness at room temperature during 8 weeks can be recommended as optimal maceration conditions for production of the aromatic carob macerate with functional properties.

Quality and Sensory Study of Fresh Sour Cherry Juices Upon Cultivar, Growing Area and Weather Conditions.

Freshly squeezed sour cherry (Prunus cerasus L.) juices of Maraska Brac (BR), Maraska MA (MA), Oblacinska 18 (18), Oblacinska Osjecka (OS), Rexelle (REX), Heimann conserved (HEI), and Kelleris 16 (KEL) grown in different climate regions (Mediterranean and continental) in two consecutive harvest years were analyzed for quality and sensory properties. Obtained results showed that investigated sources of variation (cultivar, growing area, harvest year) significantly influenced physical, chemical, and sensory properties of tested juices. All juices had high soluble solids (SS) (17.26% to 25.89%), especially BR (24.19%), MA (25.89%), 18 (24.66%), and OS (23.61%). Juices also had considerable total phenols (TP) content (2.19 to 5.87 mg GAE ml-1 ) and antioxidant capacity (AC) (0.020 to 0.045 mmol TE mL-1 ) with BR, MA, REX, and HEI containing the highest amounts of bioactive compounds (BR = 5.87 mg GAE mL-1 , MA = 5.68 mg GAE mL-1 , REX = 4.42 mg GAE mL-1 , HEI = 4.27 mg GAE mL-1 ). Sensory evaluation showed that all juices had satisfying sensory profiles with BR, MA, 18, OS, and KEL juices scoring particularly high (5.3 to 8.9 of 10) in desirable sensory properties (color intensity [CI], sour cherry taste [SCT], harmonious taste [HT], taste fullness [TF], sour cherry odor [SCO], and viscosity [V]) of fresh sour cherry cloudy juice. Juices from Mediterranean area achieved higher values for SS, pH, SS/TA ratio, TP, AC, SCT, HT, TF, SCO, and V compared to continental zone juices. Harvest year with higher temperatures and less rainfall also resulted in higher SS, pH, TP, and AC as well as stronger expression of desirable sensory properties. PRACTICAL APPLICATION: The results of this study provide an insight into sour cherry potential for use in functional food products, including today's popular smoothies, since it is fruit rich with health contributing compounds and possesses desirable sensory characteristics (color intensity, sour cherry taste, harmonious taste, taste fullness, sour cherry odor, and viscosity) at the same time, which can differ due to cultivar, climate diversity as well as weather conditions.

Influence of Carob Flour and Carob Bean Gum on Rheological Properties of Cocoa and Carob Pastry Fillings.

The aim of this study was to develop a new cocoa and carob based pastry filling and explore the influences of carob flour and carob gum on the rheological and textural properties, specifically (i) the effect of increasing the amount of carob flour and (ii) the effect of carob bean gum naturally present in the carob flour with seeds versus the commercially available carob bean gum. All samples analyzed in this study exhibited shear thinning behavior. The texture analysis revealed a significant (p < 0.01) increase in consistency and firmness in samples with higher amounts of carob flour added, while higher temperatures significantly (p < 0.01) decreased adhesiveness. When comparing naturally occurring and commercially available LBG (locust bean gum), it was concluded that lower concentrations (up to 0.45% w/w) of naturally occurring LBG work just as well at the same concentrations of commercially available LBG, but this effect cannot be confirmed for higher LBG concentrations, nor for rheological properties determined at higher temperatures (80 °C).

Polysaccharides and Antioxidants from Culinary-Medicinal White Button Mushroom, Agaricus bisporus (Agaricomycetes), Waste Biomass.

Agaricus bisporus, also known as the white button mushroom or champignon, is the most cultivated mushroom species worldwide. In addition to its favorable nutrient profile, it contains a number of compounds with antioxidant, antibacterial, anti-inflammatory, antitumor, and immunomodulatory activities. Waste biomass is a secondary product obtained from A. bisporus during the harvesting stage. It is underused, although it could be a cheap source of polysaccharides and antioxidants for use in food and feed production, or a source of nutraceuticals and cosmeceuticals. In this study, waste biomass was used as raw material for extraction of crude polysaccharides. The mean amount of crude polysaccharides extracted was 106 g/kg dry weight debris-free mushroom waste biomass. The crude polysaccharides recovered contained 11.57% α-glucan and 16.37% ß-glucan. Total carbohydrates composed 44.18%. No significant differences were found in the Fourier transform infrared spectra, which confirmed the presence of protein, α-glucan, and ß-glucan in all samples; phenols were detected only in waste biomass and market-ready A. bisporus fruiting bodies. The total phenol content in methanol extracts of waste biomass and A. bisporus fruiting bodies was 6.16 and 11.25 mg gallic acid equivalents/g extract, respectively. Antioxidant capacities of methanol extracts from waste biomass, as determined by spectrophotometric techniques, were 22.67 µmol Trolox/g extract (ABTS radical scavenging), 51.77 µmol Fe2+/g extract (ferric-reducing antioxidant power), and 51.52% (DPPH radical scavenging). Although these values were lower than those for A. bisporus fruiting bodies, the waste biomass has great potential for use in food, feed, and other bioproducts of economic importance.

Flow properties and chemical composition of carob (Ceratonia siliqua L.) flours as related to particle size and seed presence.

Due to abundance in carbohydrates, dietary fibres and bioactive compounds, as well as for its outspread and low prices, carob (Ceratonia siliqua L.) flour has a great potential of use as a functional ingredient. The aim of this study was to analyse this potential by physical and chemical properties assessment of different particle sizes of carob flour with and without seeds. The influence of seed presence on physical and chemical properties of flour was also investigated. Seed presence in carob flour led to higher cohesivity and cake strength. It also affected the extraction efficiency of polyphenols, which was confirmed by the ranking of samples according to their procyanidin and tannins contents. With regard to the carbohydrate content, significant differences (P<0.05) between the contents of fructose and glucose was established in samples differing by the presence of carob seeds. Spearman rank order correlations revealed a significant difference (P<0.05) between physical and chemical properties of carob flours. These findings confirm the importance of understanding physical and chemical properties of carob flours in order to use them efficiently as a functional food ingredient.

Assessment of Drying Characteristics and Texture in Relation with Micromorphological Traits of Carob (Ceratonia silliqua L.) Pods and Seeds.

Carob tree (Ceratonia siliqua L.) is a perennial leguminous evergreen tree native to the coastal regions of the Mediterranean basin and is considered to be an important component of vegetation for economic and environmental reasons. Two constituents of the pod, pulp and seeds, can be used as feed or in food production. In this study, drying characteristics, texture and microstructure of carob pods were studied. Three different carob samples were prepared: whole carob pod, carob pod parts and carob seed. The drying experiments and the modelling showed that carob seeds had the highest drying rate, followed by pod parts and the whole, intact carob fruit. Texture studies showed that the maximum compression force depended on the area of the carob fruit on which compression tests were performed. The seeds showed the highest compression force, followed by the stem zone, the tip and the centre of the fruit. Differences in drying behaviour and texture of carob pods can successfully be interpreted by the micromorphology of the carob pods and seeds. Determining the drying rate, maximum compressive force and micromorphological traits is of great importance for further carob processing (e.g. milling, sieving, carob bean gum production or usage in food or feed products).

Antioxidant and Antimicrobial Properties of Veronica spicata L. (Plantaginaceae).

Tea made from Veronica spicata L. (syn. Pseudolysimachion spicatum (L.) Opiz, family Plantaginaceae) herb is used in traditional medicine as expectorant for cough and throat rinsing. To get insight into chemical compounds of V. spicata, the essential oil content (analysed by GC-FID and GC-MS), the content of phenolic compounds (analysed by HPLC), the content of macroelements and trace elements (analysed by ICP-AES), quantity of total phenols and total flavonoids (analysed by UV/Vis spectrophotometer), and antioxidant and antimicrobial properties were investigated. The main compounds of the essential oil were phytol (21.13%), heptacosane (10.22%) and pentacosane (8.91%). The most abundant investigated macroelement was K (8261 mg/kg) while Fe was the most represented element (32.49 mg/kg) among investigated micronutrients. Ten phenolic compounds (chrysin, rutin, quercitrin, quercitrin, and cichoric, ferulic, protocatehuic, rosmarinic, syringic and tannic acid) were identified and quantified. Additionally, V. spicata extract demonstrated notable radical-scavenging and chelating properties. The bacterial and fungal strains used in study were found to be susceptible toward methanolic and ethyl-acetate extracts with MIC values between 1.25 and 5.00 mg/mL using microdilution method. Aquose extracts were found to be antimicrobial inactive.

Antimicrobial and Antioxidant Properties of Satureja Montana L. and S. Subspicata Vis. (Lamiaceae).

Satureja montanaL. and S. subspicata Vis. (Lamiaceae) are used for centuries in traditional medicine of Balcanic people in the healing of the lymphatic nodule and respiratory system inflammation. In this paper the amount of total phenols and flavonoids (analyzed by UV/Vis spectrophotometry), phenolic compounds profile (analyzed by HPLC), antimicrobial and antioxidant activities were studied in samples collected in seven per species populations of S. montanaand S. subspicatain Croatia. Eight phenolic compounds (rutin, quercetin, caffeic, p-coumaric, ellagic, protocatehuic, rosmarinic, and syringic acid) were identified and quantified using HPLC in methanolic and ethanolic extracts. Results showed that both species contained polyphenolics and other antioxidant compounds with chelating and radical-scavenging properties. The extracts prepared from both species showed broad spectrum of antimicrobial activity on in vitrotested microbial species (Staphylococcus aureus, Escherichia coli, Candida albicans, C. dubliniensis, C. krusei, C. glabrata, C. parapsilosis, and Microsporum gypseum).

Fortification of instant coffee beverages - influence of functional ingredients, packaging material and storage time on physical properties of newly formulated, enriched instant coffee powders.

BACKGROUND: Consumer demands for healthy, functional foods are growing rapidly nowadays. Coffee, as one of the most widespread commodities, represents an interesting aspect for enrichment, since it is consumed by millions of people on a daily basis. The aim of this study was to formulate enriched instant coffee powders with the purpose of estimating the influence of storage time, functional ingredients and packaging material on physical and sensory properties of the mixtures. RESULTS: Storage time of 6 months significantly (P <0.05) influenced moisture content of the mixtures, which rose linearly with an increase in storage time. Packaging material proved to be an important variable affecting moisture content, particle size, colour and cohesion index. Functional ingredients (vitamins A and C, iron, inulin and oligofructose) influenced particle size, dispersibility, wettability and, in terms of sensory analysis, grades for aftertaste, chemical taste and overall acceptability. CONCLUSION: Addition of functional ingredients significantly influenced some particle size distribution parameters and reconstitution properties, causing an increase in wettability and dispersibility times. Furthermore, in sensory terms, it influenced aftertaste and chemical taste grades. Packaging material significantly influenced moisture content, some particle size distribution parameters, colour and cohesion index.

Reactive Nitrogen Species Scavenging Capacity of Aqueous and Ethanolic Extracts from Galinsoga parviflora and G. quadriradiata Herbs.

The aim of the study was to determine the scavenging capacity of aqueous and ethanolic extracts derived from the herb of two species of Galinsoga against NO and ONOO-. In both tests the aqueous extracts of both Galinsoga species were more active than the ethanolic extracts. The highest scavenging activity against NO, and also against ONOO- was shown by G. quadriradiata aqueous extract (SC50 ± SD = 1.54 ± 0.32, and 2.10 ± 1.98, respectively). The study showed that the activity of the extracts from both Galinsoga species is related not only to the presence of flavonoids but also to caffeic acid derivatives and caffeic acid. The results of this study partly explain the beneficial effects of Galinsoga herb extracts in the treatment of skin diseases.

Comparison of Campylobacter jejuni pulsotypes isolated from humans and poultry in Split and Dalmatia County, Croatia.

Consumption of poultry is considered to be an important source of human infection with Campylobacter. In the period from 2008 to 2010, 50 isolates of Campylobacter jejuni from human faeces were analysed and compared with 61 isolates from poultry by pulsed field gel electrophoresis using SmaI and KpnI. Based on the analysis of SmaI macrorestriction profiles, 86 isolates (77.5 %) were assigned to 15 S clusters: 31 (62 %) from humans and 55 from poultry (90.2 %). Altogether 21 isolates (19 %) exhibited macrorestriction profiles common to both humans and poultry after restriction with SmaI and KpnI. A total of five identical pulsotypes were isolated from both poultry and patients and one of them appeared in eight different locations in the time interval of one year. These results indicate that poultry could be an important source of Campylobacter infection in Split and Dalmatia County which is the biggest County in Croatia and the most important tourist destination.

Effect of polyoxyethylene and polyoxypropylene nonionic block copolymers on performance and recruitment of immune cell subsets in weaned pigs.

BACKGROUND: Because European-wide directives are restricting the non-clinical use of antibiotics as in-feed growth promotors in swine production, there is an intensive search for alternative strategies for control and prevention of losses among young pigs. With the growing knowledge of the porcine immune system and its endogenous modulation, it has been clearly established that exogenous immunomodulation using adjuvants and immune response modifiers (IRMs) represents an important prophylactic/therapeutic approach in the prevention/treatment of both stress- and microbial-induced disorders that accompaning weaning. However, it is essential to select a fully evaluated agent which may act either as a nonspecific IRM or synergistically as an adjuvant with vaccines. The synthetic macromolecules with a long history as adjuvant and IRM are nonionic block copolymers which consist of polyoxyethylene (POE) and polyoxypropylene (POP) molecules. METHODS: The aim of this work was to evaluate the effectiveness of POE-POP given as a single peroral dose on productivity parameters such as body weight gain, feed intake and feed conversion ratio, and systemic and intestinal immune parameters by assessing the proportions of CD45+ lymphoid cells, CD4+ and CD8+ T cells, and CD21+ B cells in the peripheral blood as well as the number of CD45RA+ naive lymphoid cells residing in the ileal mucosa in weaned pigs during a follow-up study 5 weeks after the treatment. RESULTS: Pigs treated with POE-POP had better feed intake (+ 14.57%), higher average body mass at the end of the experiment (20.91 kg vs. 17.61 kg), and higher body weight gain in relation to Day 0 (191.63% vs. 144.58%) as well as in relation to nontreated pigs (+ 18.74%), with a lower feed conversion ratio (- 30.26%) in comparison to the control pigs. A much lower diarrhea severity score (5 vs. 54) was recorded in pigs treated with POE-POP (- 90.74%) than in the control pigs. A higher average diarrhea severity (ADS) was recorded in the control pigs (1.54 vs. 0.14), whereas the treatmant group had much a lower ADS ratio (- 90.91%) after 35 days of the experiment. The pigs that were treated with POE-POP had an increased proportion of CD45+, CD4+ and CD8+ cells at Day 21 (at p < 0.05, p < 0.05 or p < 0.01, respectively), Day 28 (at p < 0.01, respectively) and Day 35 (at p < 0.01, p < 0.05 or p < 0.01, respectively) as well as of CD21+ cells at Day 28 (p < 0.05) and Day 35 of the experiment (p < 0.01). Also, these pigs had more numerous CD45RA+ cells in interfollicular (p < 0.05) and follicular areas (p < 0.01) of the ileal Peyer's patches than did control pigs. CONCLUSION: This property of POE-POP to induce recruitment of circulating and intestinal immune cell subsets in weaned pigs may allow the use of IRM-active block copolymers as adjuvants for vaccines, particularly those orally delivered and targeted to the gut-associated lymphoid tissues that are well known to promote rather tolerogenic than protective immune responses.

Empiric mathematical model for predicting the content of alpha-acids in hop (Humulus lupulus L.) cv. Aurora.

The aim of this research is to find a simple mathematical model due to sum of effective temperatures and rainfalls from second germination after spring pruning till the technological maturity of hop cones, in order to achieve reliable prognosis of alpha-acids content in hop cv. Aurora. After mathematical analyses of experimental data by Eurequa Formulize 0.96 Beta software 17 equations were offered, and after substituting the values of dependent and independent variables in all equations only one equation was chosen with p = 0.034 (p<0.05). This equation is not reliable in extremely drought year if crop evapotranspiration ET0 in July is higher than 4.5, primarily because of negative influence on formation and development of hop glandular trichomes. Considering achieved results it is possible to suggest following general equation for alpha-acids accumulation in hop: y = [(k 1 w) - k 2 - (k 3 w (2))/x] ÷ (-10) ↔ ET 0 July ≤ 4.5. Where y is alpha acids content in dry matter (%), x = sum of effective temperatures and w = sum of rainfals, both from second germination after spring pruning till technological maturity of hop cones. Coefficients k 1 , k 2 and k 3 are determined for cultivar Aurora (53.8, 453 and 1.33, respectively).

Investigation of chemical compounds, antioxidant and antimicrobial properties of teucrium arduini L. (lamiaceae).

In this paper chemical composition of the essential oil (analysed by GC and GC-MS), the content of phenolic compounds (analysed by HPLC), quantity of total phenols and total flavonoids (analysed by UV/Vis spectrophotometer), antioxidant and antimicrobial activities of ethanolic extracts were investigated in endemic Teucrium arduini L. in population of Mt Biokovo (Croatia). The oil was characterized by a high concentration of sesquiterpene hydrocarbons (70.4%) of which ß-caryophyllene (35.2%) and germacrene D (18.7%) being the major compounds. Three phenolic compounds (quercetin, ferulic acid and rosmarinic acid) were identified and quantified in ethanolic extract of T. arduini using HPLC. The results also showed that T. arduini is a source of polyphenolic and other antioxidants with radical-scavenging and chelating properties. The ethanol extracts prepared from the leaf of T. arduini showed broad spectrum of antimicrobial activity on Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus brasiliensis, which are susceptible on concentration below or equal to 4.00 mg/mL, whilst Microsporum gypseum was resistant at investigated concentrations.

Chemotaxonomic and micromorphological traits of Satureja montana L. and S. subspicata Vis. (Lamiaceae).

Satureja montana and S. subspicata are used as spice, pepper substitute, for preparing tea, juice, and as a medicine. Fourteen populations (seven per species) of Satureja montana L. and S. subspicata Vis. growing in Croatia were examined to determine the chemical composition of the essential oil (analyzed by GC-FID and GC/MS), the content of macroelements (Na, K, Ca, Mg) and trace elements (B, Fe, Cu, Mn, Zn, Al, Pb, Cr, Cd, Ni, Hg, As) analyzed by ICP-AES, antioxidant compounds (analyzed by UV/VIS spectrophotometer), and the types and distribution of trichomes (analyzed by scanning electron microscopy). The main constituents of the essential oil were carvacrol and thymol in S. montana (all populations belong to one phenol chemotype), while α-eudesmol, ß-eudesmol, and spathulenol dominated in S. subspicata (three chemotypes could be distinguished). Both species possess considerably higher quantities of Ca and Mg, and moderate concentrations of K and Na, while Hg and As levels were below the limit of quantification. Non-glandular trichomes, peltate trichomes, and three types of capitate trichomes were observed on leaves, stem, calyx, and corolla.