Determination of microbiological contamination,antibacterial and antioxidant activities of natural plant hazelnut (Corylus avellana L.) pollen

The aim of our study is to determine microbial contamination, antibacterial and antioxidant activities of 14 pollen samples of Corylus avellana collected from different locations in Slovakia. Microbiological analysis was carried out in two steps: microbiological assays and studies of antibacterial activity of pollen extracts. The antimicrobial properties of pollen extracts were carried out with the disc-diffusion method. Methanol (70%), ethanol (70%) and distilled water were used for pollen extracts. Five strains of bacteria such as gram-negative (Salmonella enterica subsp. enterica CCM 3807, Escherichia coli CCM 2024, and Yersinia enterocolitica CCM 5671) and gram-positive (Staphylococcus aureus CCM 2461 and Bacillus thuringiensis CCM 19T) were tested. Antioxidant activity of pollen extracts was determined by the DPPH method. Bacterial analysis includes the determination of the total bacterial count ranged from 4.08 to 4.61 log CFU g^?1, mesophilic aerobic bacteria ranged from 3.40 to 4.89 log CFU g^?1, mesophilic anaerobic bacteria ranged from 3.20 to 4.52 log CFU g^?1, coliform bacteria ranged from 3.30 to 4.55 log CFU g^?1, yeasts and filamentous fungi ranged from 3.00 to 3.56 log CFU g^?1. Microscopic filamentous fungi Aspergillus spp., Alternaria spp., Penicillium spp., Cladosporium spp., Rhizopus spp., and Paecylomyces spp. were isolated from hazelnut pollen. Yersinia enterocolitica was the most sensitive strain among ethanolic and methanolic pollen hazelnut extracts. Staphylococcus aureus was the most sensitive strain against aqueous hazelnut pollen extracts. We determined the following sensitivity against ethanol pollen extracts respectively: Yersinia enterocolitica?>?Salmonella enterica?>?Staphylococcus aureus?>?Bacillus thuringiensis?>?Escherichia coli. Methanol pollen extracts had shown following sensitivity: Yersinia enterocolitica?>?Salmonella enterica?>?Escherichia coli?>?Staphylococcus aureus?>?Bacillus thuringiensis. Aqueous extracts had shown the following sensitivity: Staphylococcus aureus?>?Salmonella enterica?>?Escherichia coli?>?Bacillus thuringiensis?>?Yersinia enterocolitica. Hazelnut pollen extracts have over 82% antioxidant capacity in samples from non-urban zones. An elevated level of antioxidant potential in the pollen is determined by its biological properties conditioned by biologically active substances. DPPH method allowed characterizing pollen as a source of antioxidants.     

Antibacterial activity of different natural honeys from Transylvania,Romania

Honey is used in food industry and medicine due to its nutritive, therapeutic and dietetic qualities. The microbiological characteristics of 10 unpasteurized honey samples of known origin, collected from Transylvania beekeepers (Romania) were determined. The antibacterial activity of these types of honey against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Salmonella enteritidis, Salmonella anatum, Salmonella choleraesuis, Bacillus cereus, Bacillus subtilis subsp. spizizenii and Listeria monocytogenes strains was studied. The most sensitive to the antibacterial activity were the two staphylococus strains (the largest diameter of inhibition zone was 18 mm) and B. subtilis strains (13.5 mm). The strains of B. cereus, E. coli, L. monocytogenes and Salmonella spp. were found to present resistance to some of the honey samples. Manna, sunflower and polyfloral honeys presented high antibacterial activity while acacia and linden honeys had a lower activity in terms of the number of sensible strains. Statistical analysis shows that the type of strains and the type of honey have influence on the diameter of inhibition.     

Antibacterial activity against Clostridium genus and antiradical activity of the essential oils from different origin

In the present study, the antimicrobial and antiradical activities of 15 essential oils were investigated. The antimicrobial activities were determined by using agar disc diffusion and broth microdilution methods against Clostridium genus and antioxidant properties of essential oils by testing their scavenging effect on DPPH radicals activities. We determined the antibacterial activity of Clostridium butyricum, Clostridium hystoliticum, Clostridium intestinale, Clostridium perfringens and Clostridium ramosum. We obtained the original commercial essential oils samples of Lavandula angustifolia, Carum carvi, Pinus montana, Mentha piperita, Foeniculum vulgare Mill., Pinus sylvestris, Satureia montana, Origanum vulgare L. (2 samples), Pimpinella anisum, Rosmarinus officinalis L., Salvia officinalis L., Abies alba Mill., Chamomilla recutita L. Rausch and Thymus vulgaris L. produced in Slovakia (Calendula a.s., Nova Lubovna, Slovakia). The results of the disk diffusion method showed very high essential oils activity against all tested strains of microorganisms. The best antimicrobial activity against C. butyricum was found at Pimpinella anisum, against C. hystoliticum was found at Pinus sylvestris, against C. intestinale was found at Satureia hortensis L., against C. perfringens was found at Origanum vulgare L. and against C. ramosum was found at Pinus sylvestris. The results of broth microdilution assay showed that none of the essential oils was active against C. hystoliticum. The best antimicrobial activity against C. butyricum was found at Abies alba Mill., against C. intestinale was found at Abies alba Mill., against C. perfringens was found at Satureia montana and against C. ramosum was found at Abius alba and Carum carvi. Antioxidant DPPH radical scavenging activity was determined at several solutions of oil samples (50 μL.mL^?1–0.39 μL.mL^?1) and the best scavenging effect for the highest concentration (50 μL.mL^?1) was observed. The antioxidant properties were different in particular plant species. The highest% of inhibition after 30 min. of reaction was observed at Origanum vulgare (93%), Satureia montana (90.66%) and Lavandula augustifolia (90.22%).     

Effect of Cuscuta campestris parasitism on the physiological and anatomical changes in untreated and herbicide-treated sugar beet

The effects of field dodder on physiological and anatomical processes in untreated sugar beet plants and the effects of propyzamide on field dodder were examined under controlled conditions. The experiment included the following variants: N—noninfested sugar beet plants (control); I - infested sugar beet plants (untreated), and infested plants treated with propyzamide (1500 g a.i. ha^?1 (T₁) and 2000 g a.i. ha^?1(T₂)). The following parameters were checked: physiological—pigment contents (chlorophyll a, chlorophyll b, total carotenoids); anatomical -leaf parameters: thickness of epidermis, parenchyma and spongy tissue, mesophyll and underside leaf epidermis, and diameter of bundle sheath cells; petiole parameters: diameter of tracheid, petiole hydraulic conductance, xylem surface, phloem cell diameter and phloem area in sugar beet plants. A conventional paraffin wax method was used to prepare the samples for microscopy. Pigment contents were measured spectrophotometrically after methanol extraction. All parameters were measured: prior to herbicide application (0 assessment), then 7, 14, 21, 28 and 35 days after application (DAA). Field dodder was found to affect the pigment contents in untreated sugar beet plants, causing significant reductions. Conversely, reduction in the treated plants decreased 27% to 4% for chlorophyll a, from 21% to 5% for chlorophyll b, and from 28% to 5% for carotenoids (T₁). Also, in treatment T_2, reduction decreased in infested and treated plants from 19% to 2% for chlorophyll a, from 21% to 2% for chlorophyll b, from 23% to 3% for carotenoids and stimulation of 1% and 2% was observed 28 and 35 DAA, respectively. Plants infested (untreated) by field dodder had lower values of most anatomical parameters, compared to noninfested plants. The measured anatomical parameters of sugar beet leaves and petiole had significantly higher values in noninfested plants and plants treated with propyzamide than in untreated plants. Also, the results showed that propyzamide is an adequate herbicide for control of field dodder at the stage of early infestation.     

Exo-pectinase production by Bacillus pumilus using different agricultural wastes and optimizing of medium components using response surface methodology

In this research, the production of exo-pectinase by Bacillus pumilus using different agricultural wastes was studied. Agricultural wastes containing pectin such as wheat bran, sugar beet pulp, sunflower plate, orange peel, banana peel, apple pomace and grape pomace were tested as substrates, and activity of exo-pectinase was determined only in the mediums containing sugar beet pulp and wheat bran. Then, effects of parameters such as concentrations of solid substrate (wheat bran and sugar beet pulp) (A), ammonium sulphate (B) and yeast extract (C) on the production of exo-pectinase were investigated by response surface methodology. First, wheat bran was used as solid substrate, and it was determined that exo-pectinase activity increased when relatively low concentrations of ammonium sulphate (0.12–0.21 %?w/v) and yeast extract (0.12–0.3 %?w/v) and relatively high wheat bran (~5–6 %?w/v) were used. Then, exo-pectinase production was optimized by response surface methodology using sugar beet pulp as a solid substrate. In comparison to P values of the coefficients, values of not greater than 0.05 of A and B ² showed that the effect of these process variables in exo-pectinase production was important and that changes done in these variables will alter the enzyme activity.     

Fumaric acid production using renewable resources from biodiesel and cane sugar production processes

The microbial production of fumaric acid by Rhizopus arrhizus NRRL 2582 has been evaluated using soybean cake from biodiesel production processes and very high polarity (VHP) sugar from sugarcane mills. Soybean cake was converted into a nutrient-rich hydrolysate via a two-stage bioprocess involving crude enzyme production via solid state fermentations (SSF) of either Aspergillus oryzae or R. arrhizus cultivated on soybean cake followed by enzymatic hydrolysis of soybean cake. The soybean cake hydrolysate produced using crude enzymes derived via SSF of R. arrhizus was supplemented with VHP sugar and evaluated using different initial free amino nitrogen (FAN) concentrations (100, 200, and 400 mg/L) in fed-batch cultures for fumaric acid production. The highest fumaric acid concentration (27.3 g/L) and yield (0.7 g/g of total consumed sugars) were achieved when the initial FAN concentration was 200 mg/L. The combination of VHP sugar with soybean cake hydrolysate derived from crude enzymes produced by SSF of A. oryzae at 200 mg/L initial FAN concentration led to the production of 40 g/L fumaric acid with a yield of 0.86 g/g of total consumed sugars. The utilization of sugarcane molasses led to low fumaric acid production by R. arrhizus, probably due to the presence of various minerals and phenolic compounds. The promising results achieved through the valorization of VHP sugar and soybean cake suggest that a focused study on molasses pretreatment could lead to enhanced fumaric acid production.     

Low dose exposure of patulin and protective effect of epicatechin on blood cells in vitro

In the present study, we aimed to assess antioxidant status in erythrocytes in vitro after patulin (PAT) and epicatechin exposure by measuring antioxidant enzymes (superoxide-dismutase – SOD, glutathione peroxidase – GPx and catalase – CAT) and parameters associated with oxidative stress (malondialdehyde – MDA and ROS). We also investigated the effect of PAT on viability and count of lymphocytes and lymphocyte subpopulations in rabbit blood in vitro. Whole blood of rabbits was used for analysis of antioxidant changes in rabbit erythrocytes after epicatechin and PAT treatment (separately or in combination, at concentrations of 0.2; 2; 20; 200?µg mL^–1 of epicatechin and 0.5; 5; 10?µg mL^–1 of PAT). Whole blood of rabbits was also used for analysis of count and viability of lymphocytes after PAT treatment at concentrations of 10; 25 and 50?µg mL^–1. Results from our experiment confirmed the ability of epicatechin to protect cells against oxidative stress and lipoperoxidation. Our findings indicate that mycotoxin PAT in low concentrations did not affect the activity of antioxidant enzymes in erythrocytes of rabbits significantly. Only slight non-significant changes in lymphocytes count after treatment with low doses of PAT in rabbit blood were observed.     

Antioxidant and metabolism responses to polyphenol stress in cyanobacterium Microcystis aeruginosa

Three common polyphenol compounds Gallic Acid (GA), Pyrogallic Acid (PA) and Catechol (CA) are known to have allelochemical-exhibiting inhibitory effects on the growth of the cyanobacterium Microcystis aeruginosa (M. aeruginosa). Metabolism and antioxidant responses in M. aeruginosa were investigated to elucidate the mechanism by which the three polyphenols inhibit algal growth. The inhibition effects of polyphenols were in the order of CA > PA > GA. The GA and CA exposures increased protein contents, superoxide dismutase (SOD) activity, catalase (CAT) activity and soluble sugar, especially for exposure to GA of 25 mg L^?1. Soluble sugar content increased significantly especially when exposed to CA for 72 h. When exposed to PA, protein content, and SOD and CAT activities initially increased but over longer treatment time the activities decreased, in contrast to sugar content. Our results suggest that PA exposure for longer periods of time may inhibit catabolism action, while CA exposure could induce more oxide stress than GA or PA. The overall study showed that polyphenol-induced oxidative damage might be responsible for polyphenol inhibition on M. aeruginosa growth. The increases in cellular antioxidant enzymes and soluble sugar may have been to counteract the oxidative stress.     

Visible light photocatalytic antibacterial activity of Ni-doped and N-doped TiO<Subscript>2</Subscript> on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> bacteria

The Ni-doped and N-doped TiO₂ nanoparticles were investigated for their antibacterial activities on Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) bacteria. Their morphological features and characteristics such as particle size, surface area, and visible light absorbing capacity were compared and discussed. Scanning electron microscopy, X-ray diffraction, and UV–visible spectrophotometry were used to characterize both materials. The inactivation of E. coli (as an example of Gram-negative bacteria) and S. aureus (as an example of Gram-positive bacteria) with Ni-doped and N-doped TiO₂ was investigated in the absence and presence of visible light. Antibacterial activity tests were conducted using undoped, Ni-doped, and N-doped TiO₂. The N-doped TiO₂ nanoparticles show higher antibacterial activity than Ni-doped TiO₂. The band gap narrowing of N-doped TiO₂ can induce more visible light absorption and leads to the superb antibacterial properties of this material. The complete inactivation time for E. coli at an initial cell concentration of 2.7?×?10⁴ CFU/mL was 420 min which is longer than the 360 min required for S. aureus inactivation. The rate of inactivation of S. aureus using the doped TiO₂ nanoparticles in the presence of visible light is greater than that of E. coli. The median lethal dose (LD50) values of S. aureus and E. coli by antibacterial activity under an 18-W visible light intensity were 80 and 350 mg/ml for N-doped TiO₂, respectively.     

Production of sugarcane bagasse-based activated carbon for formaldehyde gas removal from potted plants exposure chamber

Agricultural wastes such as rice straw, sugar beet, and sugarcane bagasse have become a critical environmental issue due to growing agriculture demand. This study aimed to investigate the valorization possibility of sugarcane bagasse waste for activated carbon preparation. It also aimed to fully characterize the prepared activated carbon (BET surface area) via scanning electron microscope (SEM) and in terms of surface functional groups to give a basic understanding of its structure and to study the adsorption capacity of the sugarcane bagasse-based activated carbon using aqueous methylene blue (MB). The second main objective was to evaluate the performance of sugarcane bagasse-based activated carbon for indoor volatile organic compounds removal using the formaldehyde gas (HCHO) as reference model in two potted plants chambers. The first chamber was labeled the polluted chamber (containing formaldehyde gas without activated carbon) and the second was taken as the treated chamber (containing formaldehyde gas with activated carbon). The results indicated that the sugarcane bagasse-based activated carbon has a moderate BET surface area (557 m²/g) with total mesoporous volume and microporous volume of 0.310 and 0.273 cm³/g, respectively. The prepared activated carbon had remarkable adsorption capacity for MB. Formaldehyde removal rate was then found to be more than 67% in the treated chamber with the sugarcane bagasse-based activated carbon. The plants’ responses for this application as dry weight, chlorophyll contents, and protein concentration were also investigated.

Implications: Preparation of activated carbon from sugarcane bagasse (SCBAC) is a promising approach to produce cheap and efficient adsorbent for gas pollutants removal. It may be also a solution for the agricultural wastes problems in big cities, particularly in Egypt. MB adsorption tests suggest that the SCBAC have high adsorption capacity. Formaldehyde gas removal in the plant chambers indicates that the SCBAC have potential to recover volatile gases. The results confirmed that the activated carbon produced from sugarcane bagasse waste raw materials can be used as an applicable adsorbent for treating a variety of gas pollutants from the indoor environment.     

Moist Olive Husks Addition to a Silty Clay Soil: Influence on Microbial and Biochemical Parameters

To investigate the effects of moist olive husks (MOH-residues) on soil respiration, microbial biomass, and enzymatic (o-diphenoloxidase, β-glucosidase, dehydrogenase and alkaline phosphatase) activities, a silty clay soil was incubated with 0 (control), 8 × 10³ (D), 16 × 10³ (2D) and 80 × 10³ (10D) kg ha^?1 of MOH-residues on a dry weight basis. Soil respiration and microbial biomass data indicated that the addition of MOH-residues strongly increased microbial activity proportionally to the amounts added. Data of qCO₂ suggested that the respiration to biomass ratio of the microbial population was strongly modified by MOH-residues additions during the first 90 days of incubation. The qCO₂ data suggested a low efficiency in energy yields from C oxidation during the first 2 months of soil incubation. qFDA seemed to be relatively unaffected for treatments D and 2D as compared to the control, but was significantly lowered by the application of 10D, showing the lowest hydrolytic activity of microbial biomass in this treatment up to 360 days of incubation.

o-Diphenoloxidase activity was delayed, and this delay was extended with the addition of larger quantities of MOH-residues. Alkaline phosphatase, β-glucosidase and dehydrogenase activities were in line with the findings on microbial biomass changes and activities. The biological and biochemical data suggest that the addition of a large quantity of MOH-residues (80 × 10³ kg ha^?1) strongly modifies the soil characteristics affecting the r- and K-strategist populations, and that these changes last for at least the 360 days of incubation. The data also suggest that application rates exceeding 16 × 10³ kg ha^?1 are not recommended until the agro-chemical and -physical functions of the soil are further studied.     

Co-digestion of sewage sludge and dewatered residues from enzymatic hydrolysis of sugar beet pulp

Sugar beet pulp residues (SBPR) from hydrolysis and dewatering of beet pulp were co-digested with municipal sewage sludge (MSS). The highest biogas yields of nearly 512 dm³/kg VS_fed (volatile solids fed) were achieved for SBPR, treated both as the monosubstrate and as a mixture with MSS (1 : 1 by weight). Simultaneously, the highest methane production of 348 dm³ CH₄/kg VS_fed was determined when the sewage sludge was co-digested with 35% SBPR. The analysis of digestate showed that neither ammonia nor volatile fatty acids destabilized the biogas production.

Implications:?Processing of sugar beet pulp into bioethanol via enzymatic hydrolysis and microbial fermentation has become increasingly attractive. However, in this process, only the liquid fraction derived from hydrolysis is subjected into alcoholic fermentation, whereas the remaining solid fraction needs to be utilized. This study demonstrated that sugar beet pulp residues after bioethanol production can successfully be co-digested with sewage sludge to increase biogas productivity of anaerobic digesters located at wastewater treatment plants.     

Trypsin inhibitor,antioxidant and antimicrobial activities as well as chemical composition of potato sprouts originating from yellow- and colored-fleshed varieties

Abstract

Potato sprouts could be a valuable resource of phytochemicals such as secondary plant metabolites, potential antioxidants and nutritive compounds. In this work, potato sprout extracts of five varieties were examined; they differed in major glycoalkaloid content, trypsin inhibitor activity, total polyphenol content and antioxidant activity, as well as in antimicrobial activity against Gram?+?and G???bacteria, and yeast. Sprouts of colored-fleshed tubers were characterized by higher trypsin inhibitor activity than sprouts of yellow potatoes. The strongest microorganism growth inhibition effect was observed for macerate with sprouts from the purple-fleshed Blaue Annelise variety against B. subtilis, whereas C. albicans yeasts were sensitive to macerates with sprouts from purple-fleshed Blue Congo and yellow-fleshed Vineta potato varieties.     

The in vitro effect of selected essential oils on the growth and mycotoxin production of Aspergillus species

The aim of the present study was to assess the antifungal and anti-toxinogenic activity of 15 essential oils (EOs) against three fungi of the genus Aspergillus (A. parasiticus KMi-227-LR, A. parasiticus KMi-220-LR and A. flavus KMi-202-LR). The minimum inhibitory doses (MIDs) of the tested essential oils and their antifungal activity were determined using the micro-atmosphere method. The original commercial essential oil samples of Jasminum officinale L., Thymus vulgaris L., Syzygium aromaticum (L.) Merrill &; Perry, Rosmarinus officinalis L., Ocimum basilicum L., Eucalyptus globulus Labill., Salvia officinalis L., Citrus limon (L.) Burm, Origanum vulgare L., Lavandula angustifolia Mill., Carum carvi L., Citrus sinensis (L.) Osbeck., Zingiber officinalis Rosc., Mentha piperita L. and Cinnamomum zeylanicum Nees. (C. verum J.S.Presl.) were produced in Slovakia (Calendula a.s., Nová ?ubovňa, Slovakia). All essential oils exhibited activity against all tested strains of fungi. After 14 days of incubation, A. flavus (KMi-202-LR) showed the highest susceptibility with a growth inhibition percentage (GIP) of 18.70% to C. limon and 5.92% to C. sinensis, while A. parasiticus (KMi-220-LR) exhibited a GIP of 20.56% to J. officinale. The minimum inhibitory doses (MIDs) of EOs with the most significant activity were recorded. The best antifungal activity, using the micro-atmosphere method was found in S. aromaticum with an MID of 62.5 μL L^?1 air, T. vulgaris (MID of 62.5 μL L^?1 air) and O. vulgare (MID of 31.5 μL L^?1 air) against all tested strains. Mycotoxin production of the tested strains was evaluated by the thin layer chromatography (TLC) method. Mycotoxin production of AFB₁ and AFG₁ was inhibited following all treatments with C. carvi, R. officinale and S. officinale, Eucalyptus globulus L. and O. basilicum L. Essential oils exhibited a potential inhibition activity against toxic fungi, although, these affected only the production of AFB₁.     

Microwave synthesis,characterization, and bio-efficacy of novel halogenated Schiff bases

A new series of halogenated Schiff bases was synthesized by the condensation of 5-fluoro-2-hydroxy acetophenone and 3,5-dichloro-2-hydroxy acetophenone with different alkyl amines, namely propyl, pentyl, hexyl, heptyl, octyl, nonyl, dodecyl, tetradecyl, hexadecyl, and octadecyl amines, under microwave irradiation. Newly formed molecules were characterized by Infrared and nuclear magnetic resonance (¹H NMR and ¹³C NMR) spectroscopic techniques. Further, the Schiff bases were screened for antifungal bioassay, and the results showed potential fungicidal activity against two very important plant infecting fungi, viz. Rhizoctonia solani and Sclerotium rolfsii. Among the screened compounds, 2,4-dichloro-2-[1-(propylimino)ethyl]phenol was found to be the most active compound against both R. solani (ED₅₀ 8.02 mg L^?1) and S. rolfsii (ED₅₀ 21.51 mg L^?1) followed by 2,4-dichloro-2-[1-(pentylimino) ethyl]phenol (ED₅₀ 13.02 and 29.57 mg L^?1, respectively). The synthesized compounds were also screened for antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl (DPPH)-free radical scavenging technique. All the compounds showed very low to moderate activity as compared with Gallic acid.     

Synthesis and characterization of antibacterial bio-nano films for food packaging

This study prepared antibacterial nanocomposite films for food packaging from Montmorillonite, which was modified by quaternary ammonium salts such as cetyltrimethylammonium bromide (CT), hexadecyl-tributyl phosphonium bromide (HD) and corn starch (CS). After this, it determined the antimicrobial activity of CS nanofilms against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) bacteria. Dispersion of silicate layers and starch nanocomposite films was analyzed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM), and the results indicated that presence of quaternary ammonium salts enhanced clay dispersion, and the starch films incorporated with quaternary ammonium salts would provide potential use in food packaging as nanostructural materials. The nanofilms that were obtained based on the results of the antibacterial analysis were confirmed to have much stronger antibacterial properties than those in similar studies in the literature.     

Structure and antioxidant activity of phenolic compounds isolated from the edible fruits and stem bark of Harpephyllum caffrum

Antioxidant activity in edible fruits is an important characteristic in the choice of fruits for human consumption, and has profound influence on nutrition and health. Two pharmacologically active triterpenoids, β-sitosterol and lupeol, and the powerful flavan-3-ol antioxidant, (+)-catechin, were isolated from the edible fruits of Harpephyllum caffrum while a mixture of cardanols, an alkyl p-coumaric acid ester, and (+)-catechin were isolated from the stem bark. This is the first report of these compounds being isolated from this plant. The antioxidant capacity of (+)-catechin was higher than the other isolated compounds as well as the known antioxidant, ascorbic acid.     

Synthesis and antifungal activity of 2H-1,4-benzoxazin-3(4H)-one derivatives

A series of 2-alkyl-2H-1,4-benzoxazin-3(4H)-ones (4a-l) was easily synthesized by two-step process involving O-alkylation of 2-nitrophenols with methyl 2-bromoalkanoates and next “green” catalytic reductive cyclization of the obtained 2-nitro ester intermediates (3a-l). Further, 6,7-dibromo (5a-c) and N-acetyl (6) derivatives were prepared by bromination and acetylation of unsubstituted 2-alkyl-2H-1,4-benzoxazin-3(4H)-ones (4a-c). The novel compounds (3a-l, 4d-l, 5a-c and 6) were fully characterized by spectroscopic methods (MS, ¹H and ¹³C NMR). 2-Alkyl-2H-1,4-benzoxazin-3(4H)-ones (4a-l, 5a-c and 6) were screened for antifungal activity. Preliminary assays were performed using two methods: in vitro against seven phytopathogenic fungi—Botrytis cinerea, Phythophtora cactorum, Rhizoctonia solani, Phoma betae, Fusarium culmorum, Fusarium oxysporum and Alternaria alternata—and in vivo against barley powdery mildew Blumeria graminis. The tested compounds displayed moderate to good antifungal activity at high concentration (200 mg L^?1). The most potent compounds were 2-ethyl-2H-1,4-benzoxazin-3(4H)-one (4a), 2-ethyl-7-fluoro-2H-1,4-benzoxazin-3(4H)-one (4g) and 4-acetyl-2-ethyl-2H-1,4-benzoxazin-3(4H)-one (6), which completely inhibited the mycelial growth of seven agricultural fungi at the concentration of 200 mg L^?1 in the in vitro tests. Moreover, 2-ethyl-2H-1,4-benzoxazin-3(4H)-one (4a) and 4-acetyl-2-ethyl-2H-1,4-benzoxazin-3(4H)-one (6) were also screened for antifungal activity at concentrations of 100 mg L^?1 and 20 mg L^?1. In the concentration of 100 mg L^?1, the N-acetyl derivative (6) completely inhibited the growth of three strains of fungi (F. culmorum, P. cactorum and R. solani), while 2-ethyl-2H-1,4-benzoxazin-3(4H)-one (4a) completely inhibited only R. solani strain. At the concentration of 20 mg L^?1, compound 6 showed good activity only against P. cactorum strain (72%).     

Effect of carbaryl (carbamate insecticide) on acetylcholinesterase activity of two strains of Daphnia magna (Crustacea,Cladocera)

The present study was designed to investigate the effect of carbaryl (carbamate insecticide) on the acetylcholinesterase activity in two strains (same clone A) of the crustacean cladoceran Daphnia magna. Four carbaryl concentrations (0.4, 0.9, 1.8 and 3.7 µg L^?1) were compared against control AChE activity. Our results showed that after 48 h of carbaryl exposure, all treatments induced a significant decrease of AChE activities whatever the two considered strains. However, different responses were registered in terms of lowest observed effect concentrations (LOEC: 0.4 µg L^?1 for strain 1 and 0.9 µg L^?1 for strains 2) revealing differences in sensitivity among the two tested strains of D. magna. These results suggest that after carbaryl exposure, the AChE activity responses can be also used as a biomarker of susceptibility. Moreover, our results show that strain1 is less sensitive than strain 2 in terms of IC50-48 h of AChE activity. Comparing the EC50-48 h of standard ecotoxicity test and IC50-48 h of AChE inhibition, there is the same order of sensitivity with both strains.     

Antioxidant activity shown by olive pomace extracts

In this study, the effects of experimental variables such as type of solvent, sample/solvent ratio, and time of extraction have been evaluated to individuate the best results in phenolic recovery by Olive Pomaces (OP) belonging to Carolea and Ottobratica cultivars. Folin–Ciocaulteu procedure and DPPH and ABTS assays were used, respectively, for total phenol quantification and total antioxidant activity of pomace extracts. The ethanol/water mixture was resulted the most efficient extraction solvent for both olive cultivars. The highest amount of phenolic compounds (171 ± 4 mg of gallic acid 100 g^?1 of dry pomace) was obtained after extraction at 120 min with 2:1 solvent/OP (v/w) of Ottobratica Olive Pomace. The recovery of phenol compounds from olive wastes increases the sustainability of sector, allowing obtaining an extract that could be a suitable alternative in the food industry to the use of synthetic antioxidants in order to improve the quality of foods.