Assessment of anthocyanins in grape (Vitis vinifera L.) berries using a noninvasive chlorophyll fluorescence method

Anthocyanins (Anths) in grape (Vitis vinifera L.) berries harvested at véraison from Pinot Noir and Pinot Meunier cultivars were assessed nondestructively by measuring chlorophyll fluorescence (ChlF) excitation spectra. With increasing Anth content, less excitation light was transmitted to the deeper Chl layers, and thus the ChlF signal decreased proportionally. By applying Beer-Lambert's law, the logarithm of the ratio between the fluorescence excitation spectra (log FER) from a green and a red berry gave the in vivo absorption spectrum of Anths, which peaked at about 540 nm. Absolute quantitative nondestructive determination of Anths for each berry was obtained by the log FER calculated for two excitation wavelengths, 540 and 635 nm (absorbed and not-absorbed by Anths, respectively) of ChlF at 685 nm. Over a range of skin colors going from green to purple, the relationship between the log [ChlF(635)/ChlF(540)] and the Anth concentration of berry extracts was fairly well fitted (r 2 = 0.92) using a power function. Reflectance spectra on the same berry samples were also measured, and Anth reflectance indices, which were originally developed for apples and table grapes, were derived. The log FER Anth index was superior to the reflectance-ratio-based index, but was as good as the color index for red grapes (CIRG) calculated from the whole visible reflectance spectrum. The proposed log FER method, applied by means of suitable portable devices, may represent a new, rapid, and noninvasive tool for the assessment of grape phenolic maturity in vineyards.     

Antioxidant,Antimicrobial and Genotoxicity Screening of Hydro-alcoholic Extracts of Five Serbian <Emphasis Type="Italic">Equisetum</Emphasis> Species

The hydro-alcoholic extracts of five Equisetum species, E. arvense L., E. sylvaticum L., E. fluviatile L., E. palustre L. and E. telmateia Ehrh., growing-wild in Serbia were evaluated for their genotoxicity, antimicrobial activity, antioxidant capacity and the results related to the total phenol content and HPLC flavonoid profiles. The total phenol content was 92–349 μmol expressed as equivalents of chlorogenic acid per g of dried plant material. Main identified compounds were kaempferol-, quercetin- glycosides and caffeic acid derivatives. E. telmateia extract showed the greatest antioxidant capacity. Almost all tested microorganisms demonstrated some degree of sensitivity to the examined extracts. All tested extracts at 62.5 μg/ml showed higher incidence of micronucleus formation than in the control sample. The obtained data allowed mutual comparison of examined species and their assessment as possible sources of antioxidants, antimicrobials and/or genotoxic substances.     

Quantifying current and future risks of invasiveness of non-native aquatic species in highly urbanised estuarine ecosystems—A case study of the River Neretva Estuary (Eastern Adriatic Sea: Croatia and Bosnia–Herzegovina)

Biological invasions are a major driver of global environmental change as invasive non-native species can exert severe environmental impacts on invaded ecosystems. Estuaries are especially vulnerable to biological invasions, which in highly urbanised areas are further facilitated by introduction pathways linked to commercial activities. This study provides a risk screening of non-native invasive species for the highly urbanised River Neretva Estuary (Eastern Adriatic Sea: Croatia and Bosnia–Herzegovina). In total, 12 species of non-native aquatic organisms were identified and screened for their invasiveness with the Aquatic Species Invasiveness Screening Kit. Of these species, eight were classified as carrying a high risk of invasiveness under current climate conditions and nine under future climate conditions. Amongst the high-risk species, blue crab Callinectes sapidus has already caused impacts in the risk assessment area, where it also represents an important economic resource. The “horizon” species Pacific oyster Crassostrea gigas and Manila clam Ruditapes philippinarum also carried a high risk of invasiveness, which for the latter species applied under predicted global warming. The present findings will contribute towards preventative management and control measures for the conservation of the natural ecosystem of the River Neretva Estuary whilst accounting for aquatic farming demands.     

Phytochemical Properties of Satureja kitaibelii,Potential Natural Antioxidants: a New Insight

Plant Foods for Human Nutrition - Satureja kitaibelii Wierzb. ex Heuff. has a great importance in Serbian ethnopharmacology/herbal traditional medicine, as well as a flavoring food additive....     

Scoring of the extent and intensity of carp (Cyprinus carpio) skin changes made by cormorants (Phalacrocorax carbo sinensis): relationship between morphometric and histological indices

Cormorants cause damage to fish in aquaculture not only by consuming large number of fish but also by wounding them. Quantification of the actual damage in fish induced by wounding is important for reliable estimations of the overall loss in aquaculture caused by cormorants. In this study, specimens of mirror and scaly carp (Cyprinus carpio L.) injured by attacks of great cormorant (Phalacrocorax carbo L.) were collected from the ??E?ka?? fish farm in Serbia. Surfaces of superficial and open subepithelial wounds (expressed as a percent of the total body surface), estimated by computer-assisted image analysis, had mean values 10.66?±?1.86 and 0.85?±?0.90, respectively. Histological evaluation of wounds was performed by two scoring systems: system proposed by Bernet et al. (J Fish Dis 22:25?C34, 1999) and modified system described by Wahli et al. (Aquaculture 225:371?C386, 2003). The most frequent histopathological changes were disturbed epidermal structure and surface cell morphology and increased number of club cells (on more than 75% of specimens); increased number of melanocytes and their distribution in different layers in dermis; and enlarged hypodermis (more than 80% of samples) and myolysis (50% of samples). Comparison of wounds?? surfaces (surface of superficial injures) and histopathological changes, estimated by two scoring systems, revealed significant positive correlation. Bernet??s scoring system showed higher correlation coefficients, when compared with the modified Wahli??s scoring system.     

Antioxidant activity of Filipendula hexapetala flowers

The antioxidant activity of the methanol extract of Filipendula hexapetala flowers was assessed by the assay for ferric-reducing antioxidant power (FRAP), the assay for DPPH free radical scavenging ability (DPPH) and the assay for the influence of lipid peroxidation in liposomes, induced by Fe(2+)/ascorbate system and measured by the TBA test (LP). The activity of the investigated extract in all test-systems was found to be significant. The principal constituent responsible for the observed effects was isolated and identified as spiraeoside.     

Novel Spray Dried Glycerol 2-Phosphate Cross-Linked Chitosan Microparticulate Vaginal Delivery System—Development,Characterization and Cytotoxicity Studies

Chitosan microparticulate delivery systems containing clotrimazole were prepared by a spray drying technique using glycerol 2-phosphate as an ion cross-linker. The impact of a cross-linking ratio on microparticle characteristics was evaluated. Drug-free and drug-loaded unmodified or ion cross-linked chitosan microparticles were examined for the in vitro cytotoxicity in VK2/E6E7 human vaginal epithelial cells. The presence of glycerol 2-phosphate influenced drug loading and encapsulation efficacy in chitosan microparticles. By increasing the cross-linking ratio, the microparticles with lower diameter, moisture content and smoother surface were observed. Mucoadhesive studies displayed that all formulations possessed mucoadhesive properties. The in vitro release profile of clotrimazole was found to alter considerably by changing the glycerol 2-phosphate/chitosan ratio. Results from cytotoxicity studies showed occurrence of apoptotic cells in the presence of chitosan and ion cross-linked chitosan microparticles, followed by a loss of membrane potential suggesting that cell death might go through the mitochondrial apoptotic pathway.     

Nutritional management of gastrointestinal disease

The gastrointestinal (GI) tract is primarily responsible for acquiring and digesting food, absorbing nutrients and water, and expelling wastes from the body as feces. A proper diet and normally functioning GI tract are integral for the delivery of nutrients, prevention of nutrient deficiencies and malnutrition, repair of damaged intestinal epithelium, restoration of normal luminal bacterial populations, promotion of normal GI motility, and maintenance of normal immune functions (eg, both tolerance and protection from pathogens). The amount of food, its form, the frequency of feeding, and the composition of diet each have important effects on GI function and may be used to help ameliorate signs of GI disease. Although both nutrients and nonnutritional components of a diet are important to GI health, they also may cause or influence the development of GI pathology (eg, antibiotic responsive diarrhea, inflammatory bowel disease, dietary intolerance, or sensitivity and/or allergy). The appropriate diet may have a profound effect on intestinal recovery and successful management of chronic or severe GI disease.     

Development of sperm vitrification protocols for two endangered salmonid species: the Adriatic grayling, <Emphasis Type="Italic">Thymallus thymallus</Emphasis>, and the marble trout, <Emphasis Type="Italic">Salmo marmoratus</Emphasis>

Vitrification was applied to the sperm of two endangered fish species of So?a River basin in Slovenia, the Adriatic grayling (Thymallus thymallus) and marble trout (Salmo marmoratus) following testing different cooling devices and vitrifying media. Sperm was collected, diluted in species-specific non-activating media containing cryoprotectants, and vitrified by plunging directly into liquid nitrogen without pre-cooling. Progressive motility, curvilinear velocity, and straightness of fresh and vitrified-warmed sperm were evaluated with computer-assisted sperm analysis (CASA). Fertilization trials were carried out to test the effectiveness of vitrification in the case of grayling. A protocol utilizing a glucose-based extender, 30% cryoprotectants (15% methanol + 15% propylene glycol), 1:1 dilution ratio, and droplets of 2 μl on a Cryotop as cooling device yielded the highest post-thaw motility values for both Adriatic grayling (7.5?±?6.5%) and marble trout (26.6?±?15.8%). Viable embryos were produced by fertilizing eggs with vitrified grayling sperm (hatching 13.1?±?11.7%, control hatching 73.9?±?10.4%). The vitrification protocol developed in this study can be utilized in the conservation efforts for the two species as an alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.