<Emphasis Type="Italic">Aronia melanocarpa</Emphasis> phenolics and their antioxidant activity

The purpose of the present study was to evaluate small and high molecular phenolics (tannins) and antioxidant activity of Aronia melanocarpa berries, juice and pomace in order to find new potential sources of natural antioxidants. The fruits of Aronia melanocarpa Elliot were collected in the middle of October at a plantation near Wroclaw, Poland. The pomace has a much higher content of phenolics in comparision to juice and fruits. Results showed that polymeric proanthocyanins, predominantly of (−)epicatechin, are the major class of polyphenolic compounds in chokeberry, represent 66% of fruits polyphenols. The average concentration ranged from 1578.79 mg/100 g of DW for chokeberry juice up to 8191.58 mg/100 g in pomace. The concentration of phenolic acids (chlorogenic and neochlorogenic acids) in juice was higher than in pomace. Anthocyanins in Aronia melanocarpa are second phenolic compound group and represent about 25% of total polyphenols, mixture of four different cyanidin glycosides: 3-galactoside, 3-glucoside, 3-arabinoside and 3-xyloside. The higher antioxidant activity expressed as TEAC was measured in pomace >fruit >juice.     

Antitumor activities of liposome-incorporated aqueous extracts of <Emphasis Type="Italic">Anodonta </Emphasis><Emphasis Type="Italic">woodiana</Emphasis> (Lea, 1834)

To acquire the effectiveness of oral treatments, aqueous extracts of Anodonta woodiana (HB) were incorporated into liposome and its antitumor activities evaluated in vivo. The HB-loaded liposomes (HBL) were prepared at a mean size of 14.85 μm by reverse-phase evaporation method. Referring to the maximum tolerated doses test, mice with orally administrated HBL, at a 3 g/kg body weight dosage, showed no obvious acute toxic sign. Furthermore, the tumoricidal activities of HBL against C26 murine colon carcinoma, Lewis lung tumor, human QGY hepatic carcinoma and MKN-45 gastric tumor were examined, respectively. In contrast to free HB, HBL possessed remarkable antitumor activity. Simultaneously, the effect of HBL was observed in a dose-dependent manner. For C26 murine colon carcinoma and Lewis lung tumor, the inhibitory ratios of HBL were 54.36 and 51.97% at a dose of 400 mg/kg, respectively. The suppression of tumor growth and the reduction in body weight were more pronounced in human QGY hepatic carcinoma and MKN-45 gastric tumors inoculated mice by treatment of HBL during 30 days. All these promising results implied that liposome-incorporated aqueous extracts of Anodonta woodiana had a more potential application as a natural antitumor and immunomodulator formulation.     

Production of biogenic amines by<Emphasis Type="Italic"> Morganella morganii</Emphasis>,<Emphasis Type="Italic"> Klebsiella pneumoniae</Emphasis> and<Emphasis Type="Italic"> Hafnia alvei</Emphasis> using a rapid HPLC method

The histidine decarboxylating activity and production of biogenic amines by Morganella morganii (NCIMB, 10466), Klebsiella pneumoniae (NCIMB, 673) and Hafnia alvei (NCIMB, 11999) were investigated using a rapid HPLC method. Derivatisation of the bacterial samples was carried out using benzoyl chloride. A gradient elution system was used for analysis with a mixture of acetonitrile and HPLC grade water. Bacterial strains not only produce histamine in histidine-enriched broth but also the other biogenic amines. The chromatographic results show that bacterial strains are also capable of producing spermine and spermidine in histidine-enriched broth. Bacterial ammonia production by all three strains was clearly detected since ammonia is generated during the degradation of histidine. The study demonstrates that the highest histamine production was obtained by Morganella morganii, followed by Klebsiella pneumoniae, and the lowest with the Hafnia alvei. Therefore, Morganella morganii and Klebsiella pneumoniae have strong histidine decarboxylase activity since they are prolific histamine-forming bacteria     

Isolation and structure elucidation of phenolic compounds in Chinese olive (<Emphasis Type="Italic">Canarium album</Emphasis> L.) fruit

Chinese olive (Canarium album L.), one native and well-known tropical fruit tree in the southeast of China, contain a large amount of phenolics and possess great pharmacological activities. In this study, phenolics were extracted from Chinese olive fruit using 80% (v/v) aqueous acetone, and seven phenolic compounds were isolated and purified by Polyamide column and Toyopearl HW-40 column chromatography from crude extracts. Their structures were elucidated by high performance liquid chromatography coupled to diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS), and where possible by ¹H NMR and ¹³C NMR spectrometry. Except gallic acid and hyperin, five phenolic compounds including methyl gallate, ethyl gallate, corilagin, kaempferol-3-glucoside and amentoflavone were first identified in Chinese olive.     

Modeling high pressure inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> in whole milk

The survival curves of Escherichia coli and Listeria innocua inactivated by high hydrostatic pressure (HHP) were obtained at room temperature (∼22 °C) and at five pressure levels (400, 450, 500, 550 and 600 MPa) in whole milk. These curves were described by the Weibull model and parameters of this model were reduced from two to one with slight loss of goodness-of-fit. The logarithm of the time constant parameter (δ) of the reduced Weibull model was described with respect to high pressure (P). This approach can be used to define a z _p value analogous to the modeling of the classical D value (increase in pressure that results in one log unit decrease of δ values). The development of accurate survival models under high pressure, as presented here, can be very beneficial to food industry for designing, evaluating and optimizing HHP processes as a new preservation technology.     

Investigations of the natural microflora of poppy seeds (<Emphasis Type="Italic">Papaver somniferum</Emphasis>) and hazelnut kernels (<Emphasis Type="Italic">Corylus avellana</Emphasis>) including microbiological decomposition

Fatty seeds of Papaver somniferum and Corylus avellana undergo a rapid microbial degradation after being ground. Those bacteria and fungi which are mainly responsible for the microbial decay were identified, and the most important growth and death processes were documented using crucial indicator-organisms. Additionally, an aflatoxin-screening was carried out in order to assess the possible risk-potential of food intoxication. The acid value (indicator for free fatty acids) of poppy seeds and hazelnut kernels was determined during their fermentation in order to document the decomposition of triglycerides.In this study it could be proved that initially a natural decay of oil seeds is caused by bacteria, yeasts and mould fungi. After the bacteria died in the course of time, yeasts and mould fungi dominated the germ spectrum. Bacteria taking part in the degradation were identified as varieties of Staphylococcus xylosus, Enterobacteriaceae and coliforms. Yeasts were identified as Pichia burtonii, and the mould fungi are associated with the genus Alternaria.On account of the absence of the genus Aspergillus in the spectrum of mould fungi, no aflatoxin was produced.     

Influence of<Emphasis Type="Italic"> Lactobacillus plantarum</Emphasis> on<Emphasis Type="Italic"> Staphylococcus aureus</Emphasis> growth in a fresh vegetable model system

The influence of a Lactobacillus plantarum (B₄) on the growth of Staphylococcus aureus (Sa₄) was verified by impedometric methods in a suitable model reproducing the characteristics of fresh vegetables. The inoculum size of the single strains and their growth temperature were varied according to a Central Composite Design. The results obtained via statistical analysis showed that the temperature affected the growth of both S. aureus and L. plantarum strains. The pathogenic strain, independently of its inoculum size, was inhibited by L. plantarum at all the tested temperatures. A proper combination of specific lactic acid bacteria and storage temperature should improve the safety of the vegetable products.     

Efficiency of pulse pressure treatment for inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> in whole milk

Escherichia coli ATCC 11775 and Listeria innocua ATCC 33090 in whole milk were inactivated by single- and multi-pulsed (up to 10 pulses) high hydrostatic pressure (HHP) treatments. Both bacteria showed similar resistance to single- and multi-pulsed HHP. The efficiency of pulsed pressure treatment depends on the combination of holding time of each pulse and number of pulses. It was observed that multi-pulsed pressure treatment instead of traditional single-pulsed HHP could be used to pasteurize milk at a lower pressure level. Nevertheless, an optimization is necessary between the pulse holding time, number of pulses, and pressure levels to reach the desirable log-reduction of microorganisms compatible with industrial application. This study was partly presented in Joint 21st AIRAPT and 45th EHPRG International Conference on High Pressure Science and Technology held September 17–21, 2007 in Catania (Italy).     

Identification of a new phenolic compound from Chinese olive (<Emphasis Type="Italic">Canarium album</Emphasis> L.) fruit

Chinese olive (Canarium album L.), a native and a well-known tropical fruit tree in the southeast of China, contains large amount of phenolics and possesses great pharmacological activities. In this study, phenolics were extracted from Chinese olive fruit pulp using 80% (v/v) aqueous acetone, and acetone extracts were further fractioned with petroleum ether, ethyl acetate and n-butanol sequentially. From n-butanol fraction, a new phenolic compound was isolated and purified through AB-8 adsorption resin column chromatography, polyamide column chromatography and TSK Toyopearl HW-40 (S) column chromatography, and the structure of the new compound was established as 3-O-galloyl quinic acid butyl ester by electrospray ionization mass spectrometry (ESI-MS), 1D- and 2D-NMR (DEPT, COSY, HMBC, HMQC) and UV–vis techniques.     

Lactic acid fermentation of<Emphasis Type="Italic"> Brassica rapa</Emphasis>: chemical and microbial evaluation of a typical Italian product (<Emphasis Type="Italic">brovada</Emphasis>)

Brovada is an ancient traditional product from the northeast Italian region, included in any list of typical Italian products, which is obtained by natural fermentation of turnips (Brassica rapa). Turnips are cleaned and put in vats, alternating them with a layer of grape skins. Before covering the vat a mix of water and salt or water only is added. The vegetable fermentation is a spontaneous process caused by the microorganisms present on the various components (turnip and grape skins). A total 225 lactic acid bacteria (LAB) and 63 yeasts were isolated during this natural fermentation process. The main heterofermentative population was identified as Lactobacillus spp. while the major homofermentative LAB were Pediococcus spp., Candida spp. was the main yeast present. During the first 24 h of fermentation, the pH decreased rapidly to 3.7. At different steps of fermentation the headspace aroma compounds of brovada were analyzed by solid-phase-micro-extraction/gas chromatography/flame ionization detection to define the characteristic aroma profile of this particular product.     

Seasonal changes in the fatty acids of gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>) and white sea bream (<Emphasis Type="Italic">Diplodus sargus</Emphasis>) captured in Iskenderun Bay,eastern Mediterranean coast of Turkey

Seasonal variations in the fatty acid compositions of gilthead sea bream (Sparus aurata) and white sea bream (Diplodus sargus), captured in Iskenderun Bay, Eastern Mediterranean of Turkey, were investigated. Results from studying the composition over all seasons showed that the basic saturated, monounsaturated and polyunsaturated fatty acids for gilthead sea bream and white sea bream were palmitic acid (16:0), oleic acid (18:1) and docosahexaenoic acid (DHA, 22:63). The other main fatty acids for both species were myristic acid (14:0), stearic acid (18:0), palmitoleic acid (16:1), linoleic acid (18:26) (especially in autumn and winter for gilthead sea bream), and eicosapentaenoic acid (EPA, 20:53). Gilthead sea bream and white sea bream exhibited seasonal fluctuations in their fatty acid contents. EPA ratios in gilthead sea bream in the autumn, winter, spring and summer were 5.42%, 4.69%, 5.20% and 4.27%, whereas the ratios in white sea bream in autumn, spring and summer were found to be 5.03%, 4.53% and 6.97%, respectively. DHA ratios in gilthead sea bream in autumn, winter, spring and summer were 15.37%, 14.16%, 9.51% and 7.07%, whereas the ratios in white sea bream in autumn, spring and summer were found to be 11.49%, 20.17% and 7.74%, respectively. The present study suggests that the daily consumption of either 100 g of gilthead sea bream captured in any season or 100 g white sea bream captured in spring or summer could meet peoples needs for EPA+DHA fatty acids.     

Antioxidant activity of <Emphasis Type="Italic">Dioscorea batatas</Emphasis> Decne glycoprotein

Dioscorea batatas Decne (DBD) has been traditionally used as folk medicine and health food in Korea. One glycoprotein was isolated from DBD and confirmed to have 30 kDa molecular weight. The DBD glycoprotein was tested its antioxidative activity and characterized in various chemical conditions. The DBD glycoprotein has the optimal free radical scavenging activities in acidic and neutral pH and up to 85 °C. In the M²⁺ ions (Ca²⁺, Mn²⁺, and Mg²⁺) in the presence of EDTA, the activities of DBD glycoprotein reduced, compared to DBD glycoprotein alone without metal ion. Interestingly, the results in this study indicated that the activities of DBD glycoprotein do not depend on the presence of EDTA. Interestingly, when DBD glycoprotein was treated with deactivation agents (pronase E or NaIO₄), scavenging activity of DBD glycoprotein was decreased. The anti-oxidative effects of DBD glycoprotein on hydroxyl radicals in cell-free system revealed, and the DBD glycoprotein has remarkable scavenging effects on hydroxyl radicals generated by G/GO. Furthermore, the results in this study showed that the DBD glycoprotein (200 μg/ml) significantly inhibits intracellular ROS amounts and protects from cytotoxicity in primary mouse splenocyte culture treated with GO (30 mU/ml). Therefore, we speculate that DBD glycoprotein has an antioxidative potential as one of natural antioxidants.     

Novel anticoagulant compound from fermented red alga <Emphasis Type="Italic">Pachymeniopsis elliptica</Emphasis>

Natural fermentation was tested as a method of releasing active compounds during screening for potential anticoagulant activity in three types of algae (Pachymeniopsis elliptica, Sargassum horneri, and Ulva pertusa). Freeze dried algae samples (2.5 g) were fermented by adding 75 g of sugar and 500 mL of water and thereafter kept at room temperature (25 °C) for 3 months. Activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) were measured every 2 weeks for 3 months to determine the optimum time for the highest activity. Fermented P. elliptica, (which had the highest activity) was subjected to anion exchange chromatography (DEAE-cellulose) and sepharose 4B gel permeation chromatography. The purified sample was analyzed by agarose-gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) to confirm the purification and to determine the molecular mass, respectively. The 360 μg/mL of purified compound (Mwt > 500,000 Da) had both APTT and PT activities (>1,000 s). However, at the concentrations of 180 μg/mL, purified compound and heparin showed 540 and >1,000 s APTT activity, respectively. Though, the purified compound of P. elliptica considered as a weaker anticoagulant than heparin, this purified anticoagulant polysaccharide could be considered as a good alternative source as an anticoagulant. Moreover, the technique of fermentation is an inexpensive and feasible, this purified anticoagulant polysaccharide compound could be used in pharmaceutical and biomedical industry. Further investigations need to be performed to determine the mechanism of this novel anticoagulant compound. The authors Prashani Mudika Ekanayake and Chamilani Nikapitiya contributed equally to this work.     

Genetic typification by pulsed-field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA (RAPD) of wild <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> and <Emphasis Type="Italic">Oenococcus oeni</Emphasis> wine strains

Genetic typification of 120 bacterial isolates of Lactobacillus plantarum and Oenococcus oeni from different Rioja musts and wines was performed by numerical analysis of pulsed-field gel electrophoresis (PFGE) patterns with endonuclease SfiI, and 46 of them were also studied by randomly amplified polymorphic DNA (RAPD)-PCR. A comparative study of both typification methods applied to L. plantarum and O. oeni oenological strains was performed. Bacterial species was determined both by biochemical identification methods and by specific PCR analysis. A wide variety of restriction digest patterns were detected by PFGE among L. plantarum strains (36 unrelated patterns and one closely related pattern, out of 48 isolates), as well as among O. oeni strains (18 unrelated patterns out of 72 isolates). PFGE was shown to be a suitable method for strain differentiation and to determine which strains are present in wine fermentations, with a discriminatory power to type L. plantarum and O. oeni strains higher than that of RAPD-PCR.     

Antimicrobial resistance in <Emphasis Type="Italic">Escherichia coli</Emphasis> strains isolated from organic and conventional pork meat: a comparative survey

Mean counts of Escherichia coli were determined for 54 samples of organic pork meat, and in 67 samples of conventional pork meat. Up to three E. coli strains from each sample were analysed by an agar disk diffusion assay for their resistance to ampicillin, chloramphenicol, cephalotin, doxycycline, enrofloxacin, gentamicin, nitrofurantoin, sulfisoxazole and streptomycin by the agar disk diffusion method. Results indicated that the presence of E. coli was significantly (P < 0.05) higher in organic pork meat as compared to conventional pork meat. Isolates from organic pork meat exhibited lower levels of antimicrobial resistance against ampicillin (P < 0.0001), doxycycline (P < 0.0001) and sulfisoxazole (P < 0.0001), as compared to isolates from conventional meat. Moreover, presence of multi-resistant E. coli strains was significantly (P < 0.0001) higher in conventional pork meat as compared to organic pork meat, the largest differences being observed for isolates resistant to combinations of ampicillin, sulfisoxazol and/or doxycycline. Organically-farmed pork samples showed significantly lower development of antimicrobial resistance in E. coli, thus contributing to reduce the development and spread of antimicrobial resistance among these food-borne bacteria.     

Extracts of plant cell cultures of <Emphasis Type="Italic">Lavandula vera</Emphasis> and <Emphasis Type="Italic">Rosa damascena</Emphasis> as sources of phenolic antioxidants for use in foods

Ethanolic extracts of plant cell cultures of lavender (Lavandula vera) and rose (Rosa damascena) have been examined as potential food antioxidants. The L. vera cell extract quenched the radicals Fremy’s salt, DPPH (2,2-diphenyl-1-picrylhydrazyl radical), and ABTS^·+ (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic) radical) more efficiently than the R. damascena extract. Also the L. vera extract inhibited lipid oxidation in a methyl linoleate emulsion more efficiently than the R. damascena extract. However, the L. vera extract had a prooxidative effect on the iron-based Fenton reaction in an aqueous model system. A similar effect was observed for pure rosmarinic acid, but not for the R. damascena extract. The addition of L. vera extract to minced chicken meat reduced lipid oxidation (measured as thiobarbituric acid reactive species) and the loss of α-tocopherol during cold storage after the meat was cooked. This suggests the antioxidative properties of L. vera extracts dominate in a real food system.     

Phenolic compounds in Imburana (<Emphasis Type="Italic">Amburana cearensis)</Emphasis> powder extracts

In response to the increasing demand for aged distilled spirits, wood extracts have been investigated as accelerating agents in distilled spirits. The aim of this work was to obtain and to evaluate the potential use of Amburana cearensis powder extracts as an accelerating aging agent in sugar cane distilled spirits. Wood powder was submitted to different toasting treatments using conventional drying oven and microwave oven. Instead of using the traditional reflux extraction method, the phenolic compounds were extracted by means of ultrasonic treatment using a 50% ethanolic solution with pH adjusted to 4.5. The concentration of gallic acid, 5-hydroxymethylfurfural, furfural, vanillic acid, syringic acid, vanillin, coniferaldehyde, sinapaldehyde and coumarin, determined by high performance liquid chromatography (HPLC) were used as quality parameters of powder toasting treatment and ethanolic extraction. The use of toasted wood powder combined with ultrasonic extraction showed higher yields when compared to the traditional solvent extraction method when obtaining wood extracts. The extraction method presented in this work is also less time consuming.     

Antioxidant capacity and total phenolics of <Emphasis Type="Italic">Cyphostemma digitatum</Emphasis> before and after processing: use of different assays

In the framework of standardisation of new healthy food sources, this paper aimed to study the total phenolics and the antioxidant power of Cyphostemma digitatum (Vitaceae) in water and ethanol extracts, using 96-well micro plates with BMG FLUOstar Optima micro plate reader. Total phenolics by Folin–Ciocalteu method in the water extracts were significantly lower after processing, decreasing from 1.41 ± 0.06 g GAE/100 g in the raw leaves to 0.80 ± 0.08 g GAE/100 g in the processed sample; the ethanol extract revealed the same trend with higher values, decreasing from 1.95 ± 0.03 to 1.56 ± 0.12 g GAE/100 g. The antioxidant capacity was elucidated by four methods: TEAC, DPPH, FRAP and ORAC. No or very weak correlations were found between antioxidant assays and total phenolics; this confirms that the antioxidant capacity could be attributed to other molecules. The ORAC assay proved to be more powerful than the other assays; it showed 103.3 ± 2.5 mmol/100 g Trolox equivalents in the raw leaves ethanol extract and 91.9 ± 3.0 mmol/100 g in the processed sample. ORAC assay showed the opposite for the water extract where the antioxidant capacity increased from 16.7 ± 0.2 to 41.7 ± 2.7 mmol/100 g Trolox equivalents after processing, which could be attributed to new water-soluble compounds generated in the consumed form.     

Effects of process parameters on growth and metabolism of<Emphasis Type="Italic"> Lactobacillus sanfranciscensis</Emphasis> and<Emphasis Type="Italic"> Candida humilis</Emphasis> during rye sourdough fermentation

The effects of temperature, pH, inoculum level, and NaCl on the growth and metabolism of Lactobacillus sanfranciscensis and Candida humilis in rye sourdough were determined. The temperature optima for growth of C. humilis and L. sanfranciscensis were 28 and 32 °C, respectively. Yeast growth was inhibited at 35 °C. The pH did not affect yeast growth in the range 3.5–5.5, whereas growth of L. sanfranciscensis was inhibited at pH 4.0. A NaCl concentration of 4% (flour base) inhibited growth of L. sanfranciscensis but not C. humilis. The effects of the process parameters on the formation of lactate, acetate, ethanol, and CO₂ by the organisms were generally in agreement with their effects on growth. However, decreased formation of acetate by L. sanfranciscensis was observed at 35 °C although lactate and ethanol formation were not affected. In conclusion, the study provides a rationale for the stable persistence of L. sanfranciscensis and C. humilis in traditional sourdoughs and will facilitate the optimisation of sourdough fermentations in traditional and new applications.