Effect of Thermosonic Pretreatment and Microwave Vacuum Drying on the Water State and Glass Transition Temperature in <Emphasis Type="Italic">Agaricus bisporus</Emphasis> Slices

This study aimed to understand the micromechanism of thermosonic pretreatment and microwave vacuum drying on Agaricus bisporus. The water state and glass transition temperature (T _g ) of fresh and thermosonically treated Agaricus bisporus slices during microwave vacuum drying were studied using differential scanning calorimetry (DSC), low-field nuclear magnetic resonance (LF-NMR), and magnetic resonance imaging (MRI). Results showed that four population groups were contained in the initial distribution of transverse relaxation time (T ₂) data of fresh A. bisporus slices: T ₂₁ (0.38–7.05 ms), T ₂₂ (9.33–32.75 ms), T ₂₃₁ (37.65–265.61 ms), and T ₂₃₂ (305.39–811.13 ms). Thermosonic pretreatment significantly decreased the initial free water content of A. bisporus sample but was accompanied by a sharp increase in its immobilized water. “Semi-bound water transfer” appeared during microwave vacuum drying (MVD) at moisture contents (X _w ) of 0.70 and 0.60 g/g (wet basis (w.b.)) for untreated and thermosonically treated samples, respectively. MVD caused dramatic changes in the water state and enhanced the T _g by decreasing the content and mobility of immobilized water in A. bisporus tissues. The mobility of semi-bound water for thermosonically and MVD-treated samples was higher than for MVD-untreated samples, resulting in T _g values decreasing by approximately 2–11.5 °C, but the uniformity of water distribution in thermosonic-treated and MVD-treated samples was better at X _w  ≤ 0.52 g/g (w.b.).     

Characterization of aromatic compounds and biological activities of essential oils from Tunisian aromatic plants

The aromatic compounds and biological activities of essential oils from six Tunisian aromatic plants including Artemisia herba-alba, Rosmarinus officinalis, Thymus capitatus, Mentha piperita, Ocimum basilicum and Artemisia absinthium were investigated. Hydro-distillation was used to extract essential oil from these plants. The identification of compounds from essential oils was performed using GC–MS analysis. Camphor (28.47%) was the major compound of A. absinthium essential oil. High contents of verbenone (20.99%) and camphor (19.72%) were found in R. officinalis. In the case of T. capitatus, carvacrol (81.09%), gamma terpinene (6.61%) and caryophyllene (4.87%) were identified as the major compounds. While eugenol (24.69%), linalool (18.00%) were characteristic compounds of O. basilicum essential oil, camphor (39.10%) and farnesol (14.25%) together with bornyl acetate (12.31%) were the main constituents of A. absinthium. These oils were also subjected to a screening for their antioxidant activity and essential oil from A. absinthium showed a greater antioxidant activity (IC₅₀?=?0.0063 mg/mL) compared to the standard Vitamin E (IC₅₀?=?0.019 mg/mL). The antibacterial activities of the oils against seven pathogenic strains, Pseudomonas aeruginosa, Bacillus cereus, Salmonella, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Micrococcus luteus, were tested. The highest and broadest activity was shown by M. piperita; however, Ocimum basilicum was inactive against all strains. Essential oils were also evaluated for antidiabetic and anti-acetylcholinesterase activities. The IC₅₀ values of A. herba-alba and O. basilicum against α-amylase were respectively 17.76 and 16.32 µg/mL suggesting a powerful anti-diabetic effect comparable to that of acarbose (IC₅₀?=?14.88 µg/mL). R. officinalis, M. piperita and A. absinthium exhibited an interesting acetylcholinesterase inhibition with IC₅₀ equal to 22, 24 and 58 µg/mL respectively.     

Reversed-phase HPLC determination of mangiferin,isomangiferin and hesperidin in <Emphasis Type="Italic"> Cyclopia </Emphasis>and the effect of harvesting date on the phenolic composition of <Emphasis Type="Italic"> C. genistoides</Emphasis>

A reversed-phase HPLC method for separation of polyphenols in honeybush tea (Cyclopia spp.) is presented. Separation of eriodictyol, luteolin, medicagol, formononetin, mangiferin, isomangiferin, hesperetin and hesperidin was investigated. A C12 stationary phase was required to separate mangiferin and isomangiferin. The method was used to quantify the three major polyphenols (mangiferin, isomangiferin and hesperidin) in C. genistoides, C. intermedia, C. maculata and C. sessiliflora and to study the effect of harvesting date on these compounds in two types of C. genistoides. The highest levels of the xanthones, mangiferin (3.61 g/100 g) and isomangiferin (0.54 g/100 g), and the flavanone, hesperidin (1.74 g/100 g), were found for C. genistoides (both xanthones) and C. intermedia, respectively. Cyclopia sessiliflora contained the lowest levels of mangiferin (1.04 g/100 g) and hesperidin (0.29 g/100 g). The mangiferin content of both the Overberg and West Coast types decreased with harvesting date (P <0.05). The Overberg type contained more mangiferin, but hesperidin was more prominent in the West Coast type.     

Development and characterization of reconstituted hydrogel from <Emphasis Type="Italic">Aloe vera</Emphasis> (<Emphasis Type="Italic">Aloe barbadensis</Emphasis> Miller) powder

Structural and rheological characterization of reconstituted hydrogels developed from A. vera non-fibrous alcohol insoluble residue (NFAIR) powder using different methods [viz., shaking (S), heating-shaking (HS), and heating (H)] and concentrations (viz., 0.2–1.6 %, w/v) was carried out. Functional group distribution by FTIR spectroscopy and Congo red (CR) method revealed the presence of acetylated acemannan in A. vera powder. Dynamic oscillation studies of A. vera (NFAIR) fluids at all concentrations of 0.2–1.6 %, w/v, showed gel strength in the order of H > HS > S method. However, in H method, increase in concentration from 0.2 to 1.6 %, w/v showed the conformational transition from semi-diluted solution to weak gel nature. Rheological models described the effect of heating temperatures (HT); 30–90 °C, and times (Ht); 15–60 min on viscoelastic behavior in reconstituted A. vera fluids. The reconstituted A. vera hydrogel prepared with a concentration of 1.6 %, w/v using 50 °C (HT) and 30 min (Ht) condition showed a good agreement with the Power law (storage modulus, G′) and Weak gel model (complex modulus, G*) fitted data (R² > 0.94) resulting higher viscoelastic moduli intercepts; G′₀ (71.5 Pa s ^n′), G″₀ (33.5 Pa s ^n″), lower slopes; n′ (0.22), n″ (0.06), higher network strength (A _F , 121.3 Pa s^1/z ) and number of network (z, 5.3) values. The obtained results suggested that heating at 50 °C/30 min can develop aqueous weak gel networks of A. vera with enhanced gel strength which may be utilized as a novel gelling agent for wide variety of targeted applications in food and pharmaceutical sectors.     

An approach for extraction,purification, characterization and quantitation of acylated-anthocyanins from <Emphasis Type="Italic">Nitraria tangutorun</Emphasis> Bobr. fruit

In the present study, a systematic approach for extraction, purification and analysis of acylated-anthocyanins from Nitraria tangutorun Bobr. fruit was explored. Six acylated-anthocyanins in N. tangutorun fruit were identified by HPLC-MS/MS, and a rapid and efficient HPLC-DAD method was developed to analyze the acylated-anthocyanins. Ultrasonic-assisted extraction conditions of acylated-anthocyanins were optimized using response surface methodology, extraction at 70 °C for 32 min using 70% methanol solution (0.1% HCl, v/v) rendered an extract with 80.37?±?2.66 mg/100 g of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside and 97.88?±?4.06 mg/100 g of total acylated-anthocyanins. Nine macroporous resins were investigated for preliminary purification of acylated-anthocyanins. According to the static/dynamic adsorption and desorption tests, XDA-6 macroporous resin exhibited the maximum potential for preparing acylated-anthocyanins. The purity of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside (43.30 mg/g) in purified acylated-anthocyanins was 201.89 times of that of the extract (0.21 mg/g), and the purity of total acylated-anthocyanins increased from 0.36 to 56.44 mg/g. Besides, the stability (t _1/2) of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside and total acylated-anthocyanins increased by more than five-fold after purification using XDA-6. The established methods of analysis, extraction and purification of acylated-anthocyanins were hopefully utilized in food industry.     

Antioxidant activity and acetylcholinesterase inhibition of field and in vitro grown <Emphasis Type="Italic">Musa</Emphasis> L. species

Bananas and plantains (Musa L. species) have medicinal applications against diseases such as hypoglycemia, hypertension, and neurological disorders, especially Alzheimer’s disease. The demand for these plants is growing very fast, resulting in a relatively heavy load on Musaceae genetic resources. The study evaluated and compared the acetylcholinesterase (AChE) inhibition and antioxidant activity of field and in vitro plant materials of nine accessions of Musa spp. consisting of Tropical Musa banana (TMb: TMb 106, TMb 145, TMb 8, TMb 82, TMb 55) and Tropical Musa plantain (TMp: TMp 116, TMp 24, TMp 36) from the International Institute of Tropical Agriculture and Musa sapientum (MS) from the University of Ibadan Botanical garden, Nigeria. Musa accessions were estimated onto Murashige and Skoog (MS) medium supplemented with 0.18 mg/L indole acetic acid (IAA) and 4.5 mg/L benzyl amino purine (BAP). The in vitro grown accessions behaved differently with TMb 8 having the highest average shoot length of 5.03?±?0.66 cm, and average number of leaves of 5.65?±?0.38 cm at the end of 6 weeks. Leaf extracts provide more quantity of phenolics, flavonoids and higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity than the fruit extracts. The AChEI activity of the field plants ranged from 60.54?±?0.54 to 98.46?±?0.09?% and in vitro plants from 61.88?±?0.11 to 76.60?±?0.34?% at 200 µg/mL. Crude methanol extract (CME) of the in vitro plants showed higher DPPH antioxidant activity than the field plants with IC₅₀ (extract concentration providing 50?% inhibition) values ranging from 9.57?±?0.24 to 48.37?±?0.62 µg/mL compared with CME of the field samples, which had IC₅₀ ranging from 75.86?±?1.76 to 162.20?±?3.77 µg/mL. Plant tissue culture can be a reliable alternative cultivation method for mass propagation and conservation of Musa species for the production of antioxidant and acetylcholinesterase metabolites.     

Glycerides isolated from the aerial parts of <Emphasis Type="Italic">Malva verticillata</Emphasis> cause immunomodulation effects via splenocyte function and NK anti-tumor activity

A preliminary study revealed that a 10 µg/mL n-BuOH fraction of Malva verticillata aerial parts significantly enhanced splenocyte proliferation and induced significant enhancement of natural-killer (NK) cell activity against tumor cells (YAC-1). This study was initiated to identify the principal components that exhibited these activities, and four glycerides were isolated through repeated SiO₂ and ODS column chromatography. Structures of compounds 1–4 were determined to be (2S)-1-O-palmitoyl glyceride, (2S)-1-O-stearoyl glyceride, (2S)-1-O-linolenoyl glyceride, and (2S)-1,2-di-O-linoleoyl glyceride, respectively. Compounds 1–3 showed potential immune-enhancing activity in murine splenocyte and natural-killer (NK) cells at 10 µM. In contrast, compound 4 showed weak activity, indicating the monoacyl glycerides (1–3) are more effective than diacyl glyceride (4). Also, the longer the carbon number of the fatty acid in monoacyl glyceride, the better the activity, and the monoacyl glyceride including an unsaturated fatty acid (3) is more effective than the glycerides including the saturated fatty acids (1–2).     

Response surface optimization of conditions for debittering of white mahlab (<Emphasis Type="Italic">Prunus mahaleb</Emphasis> L.) juice using polystyrene resins

In the current study, debittering of white mahlab (Prunus mahaleb L.) juice using polystyrene resins was investigated and optimized using the Box–Behnken response surface methodology. The effects of independent variables including resin type (cation exchange resin, anion exchange resin, and 50:50 cation/anion exchange resin), resin dosage (0.05, 0.075, and 0.1 g/mL) and agitation speed (50, 150, and 250 rpm) on total phenolic content, total flavonoid content, total anthocyanin content (TAC), antioxidant activity, and bitterness of white mahlab juice were studied. Effects of resin type and dosage were found to be significant (p < 0.05) for all responses. A dosage of 0.05 g/mL of the anion exchange resin and an agitation speed of 250 rpm were selected as the optimum conditions based on minimum bitterness (2.42) and maximum TAC (0.018 g/mol). We could conclude that the anionic resin can greatly reduce the intensity of bitterness and maintain the composition and characteristics of the P. mahaleb juice.     

Optimization of Extraction Parameters of Total Phenolics from <Emphasis Type="Italic">Annona crassiflora</Emphasis> Mart. (Araticum) Fruits Using Response Surface Methodology

In this study, response surface methodology was used to optimize the extraction temperature (25–75 °C) and ethanol concentration (0–70 %, ethanol/water, v/v) to maximize the extraction of total phenolic compounds (TPC) from araticum pulp. The efficiency of the extraction process was monitored over time, and equilibrium conditions were reached between 60–90 min. A second-order polynomial model was adequately fit to the experimental data with an adjusted R ² of 0.9793 (p < 0.0001) showing that the model could efficiently predict the TPC content. Optimum extraction conditions were ethanol concentration of 46 % (v/v), extraction temperature of 75 °C and extraction time of 90 min. Under the optimum conditions, the araticum pulp showed high TPC content (4.67 g GAE/100 g dw) and also high antioxidant activity in the different assays used (46.56 μg/mL, 683.65 μmol TE/g and 1593.72 μmol TE/g for DPPH IC₅₀, TEAC and T-ORAC_FL, respectively). From our extraction procedure, we successfully recovered a significantly higher amount of TPC compared to other studies in the literature to date (1.5–22-fold higher). Furthermore, TPC and antioxidant activity were present in the fruit in levels that are difficult to find in other common fruits. These results expose a potential approach for improving human health through consumption of araticum fruit.     

Saltatory Rolling Circle Amplification (SRCA): a Novel Nucleic Acid Isothermal Amplification Technique Applied for Rapid Detection of <Emphasis Type="Italic">Shigella</Emphasis> Spp. in Vegetable Salad

Shigella spp. are enteric pathogens that pose a serious threat to public health worldwide. A novel saltatory rolling circle amplification (SRCA) assay was developed to detect Shigella spp. in food targeting the ipaH gene. SRCA as an isothermal amplification method requires no expensive thermocycle instrument and could avoid electrophoresis as visualization results was successfully applied for SRCA. In order to confirm the specificity of this assay, 34 strains including 11 strains belonging to different Shigella species and 23 non-Shigella bacteria were detected with pure cultures. The sensitivity of Shigella flexneri by SRCA was evaluated using agarose gel electrophoresis, which was 7.3 × 10¹ fg/μL. In addition, the amplification results were also determined by adding the fluorochrome, SYBR Green I (1 μL of 1000×), allowing naked eye visualization of results, and the sensitivity was 7.3 × 10⁰ fg/μL. Moreover, the sensitivity of PCR was 7.3 × 10² fg/μL, showing that the sensitivity of SRCA by electrophoresis and SYBR Green I fluorescence were 10- and 100-fold higher than that of PCR, respectively. The detection limit of SRCA was also evaluated with artificially inoculated vegetable salad without enrichment, and it was 4.7 × 10² and 4.7 × 10¹ CFU/g by electrophoresis and fluorescence, respectively. The detection limit by PCR was 4.7 × 10³ CFU/g, which was 10- and 100-fold higher than that of SRCA. Therefore, SRCA is a potentially reliable tool for rapid and specific detection of Shigella in food and could be useful in underdeveloped countries with limited resources.     

Detection of Viable <Emphasis Type="Italic">Vibrio cholerae</Emphasis> Cells in Seafood Using a Real-Time Visual Loop-Mediated Isothermal Amplification Combined with Propidium Monoazide

Vibrio cholerae is an important foodborne pathogen causing severe intestinal infectious diseases that have high incidence and mortality. Almost all of rapid testing methods including immunological and molecular assays for V. cholerae are incapable of distinguishing live cells from dead ones, which may overestimate the number of bacteria and result in many false positive results. To address the problems, live cell-specific dye such as propidium monoazide (PMA) is employed. The loop-mediated isothermal amplification (LAMP) assay is a nucleic acid amplification method that is fast, specific, and sensitive. In this study, we developed a real-time visual LAMP assay using PMA dye to detect thyA gene, thereby identifying viable V. cholerae cells. The results showed that only V. cholarae strains could be detected, and there was no cross-reaction with non-V. cholarae strains. Besides, the sensitivity of the PMA-LAMP assay was 1.1 × 10² CFU/mL and the entire reaction could be accomplished within 1 h. The sensitivity was on par with that of the PMA-qPCR assay. The detection limit in different artificially inoculated samples was 5 CFU/25 g materials for the tested pathogens. In the practical test, the PMA-LAMP assay performed well in comparison with PMA-qPCR and the culture method. Hence, PMA-LAMP assay can provide a highly effective and rapid approach for detecting viable V. cholerae.     

Determination of Seven <Emphasis Type="Italic">N</Emphasis>-nitrosamines in Agricultural Food Matrices Using GC-PCI-MS/MS

The quantitative analytical methods for seven N-nitrosamines including N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosodibutylamine (NDBA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), and N-nitrosomorpholine (NMOR) were established for agricultural food matrices. Four food matrices were used for the method development: rice soup as a fatless solid matrix, apple juice as a fatless liquid matrix, corn oil as a fat-rich liquid matrix, and 20 % alcohol as an alcohol matrix. A combination of solid-supported liquid-liquid extraction (SLLE) using Extrelut NT and a solid phase extraction (SPE) using Florisil was employed for fatless matrices. For an alcohol matrix, only SLLE was used without SPE, and liquid-liquid extraction (LLE) was established for a fat-rich matrix. The extract was analyzed by gas chromatography-positive chemical ionization-tandem mass spectrometry (GC-PCI-MS/MS) using ammonia gas as an ion source. Linearity, recovery, repeatability, inter-day precision, reproducibility, and uncertainty were evaluated for method validation using four matrices. Method detection limits for all of the investigated N-nitrosamines were ranged from 0.10 to 0.18 μg/kg for the rice soup, from 0.10 to 0.19 μg/kg for the apple juice, 0.10 μg/kg for the corn oil, and from 0.10 to 0.25 μg/kg for 20 % alcohol, depending on N-nitrosamines. Established methods were applied to determine seven N-nitrosamines in some agricultural food products.     

Effects of organic acid pretreatment on microstructure,functional and thermal properties of unripe banana flour

Starch availability has been implicated in unripe matured banana (Musa species), which when processed yields flour suitable for application in low gluten and composite wheat formulations. Unripe Musa species: Williams, Luvhele, Mabonde and Muomva-red obtained from fruit bunch were pretreated with ascorbic, citric and lactic acids, processed into 50 g of flour and characterised for their functional and thermal properties. Scanning electron microscope of unripe banana flour (UBF) showed varying micrographs of flour, with polygonal for Luvhele, oval for Mabonde, elongated for Muomva-red and between polygonal and spherical for Williams. The bulk density of UBF samples was within the range of 0.66–0.84 g/mL for all organic acid pretreatment while citric acid pretreated UBF had the least browning index. Significant difference (p < 0.05) was recorded in swelling power with no significant difference in water solubility index except for Mabonde UBF. Thermal properties showed single endothermic transition for all UBF samples at various pretreatment concentration. The onset temperature (To) of UBF ranges from 49.82 to 65.59 °C, peak temperature (Tp) from 60.11 to 76.71 °C, conclusion temperature (Tc) from 70.36 to 94.16 °C and enthalpy of gelatinization (ΔH) from 2.61 to 32.24 J/g. Short amylopectin chains present in starch of UBF was attributed to low To, Tp, Tc and ΔH values recorded for Mabonde cultivar, while the contribution of heat-moisture treatment rather than organic acid pretreatment of UBF samples was attributed to different gelatinization and transition temperatures recorded for all cultivars examined.     

Properties of a fibrinolytic enzyme secreted by <Emphasis Type="Italic">Bacillus subtilis</Emphasis> JS2 isolated from saeu (small shrimp) <Emphasis Type="Italic">jeotgal</Emphasis>

Bacillus species were screened to be used as starters for jeotgals, salted and fermented Korean sea foods. A strain, JS2, showing strong fibrinolytic activity was isolated from saeu (small shrimp) jeotgal, and identified as Bacillus subtilis. Bacillus subtilis JS2 grew well at 20% (w/v) NaCl concentration. SDS-PAGE of culture supernatant from JS2 showed 3 major bands of 27, 29, and 60 kDa in size. Fibrin zymography showed that the 27 kDa band was the major fibrinolytic protein. The gene, aprEJS2, was cloned and introduced into B. subtilis WB600 using pHY300PLK. A B. subtilis transformant harboring pHYJS2 showed higher fibrinolytic activity than B. subtilis JS2. aprEJS2 was overexpressed in Escherichia coli BL21 (DE3). The optimum pH and temperature for AprEJS2 were pH 8.0 and 40 °C, respectively. Km and V_max values were determined. AprEJS2 has strong α-fibrinogenase activity and moderate β-fibrinogenase activity.     

Design of Sonotrode Ultrasound-Assisted Extraction of Phenolic Compounds from <Emphasis Type="Italic">Psidium guajava</Emphasis> L. Leaves

Psidium guajava L. has gained a special attention as health plant due to the presence of phenolic compounds. Box-Behnken design (BBD) has been applied for the extraction of target compounds from guava leaves via sonotrode ultrasound-assisted extraction (UAE). Different extraction times (5, 30, and 55 min), ratios of ethanol/water (50, 75, and 100% (v/v)), and ultrasound (US) power (80, 240, and 400 W) were tested to find their effect on the sum of phenolic compound (SPC), flavonols and flavan-3-ols via HPLC-ESI-QqQ-MS, and antioxidant activity (DPPH and TEAC assays). The best process conditions were as follows: 40 min, 60% ethanol/water (v/v), and 200 W. Established method has been used to extract phenolic compounds in two guava leaves varieties (pyrifera and pomifera). Pyrifera var. showed greater values of the SPC via HPLC-ESI-QqQ-MS (49.7 mg/g leaf dry weight (d.w.)), flavonols (12.51 mg/g d.w.), flavan-3-ols (7.20 mg/g d.w.), individual phenolic compounds, and antioxidant activity (8970 ± 5 and 465 ± 6 μmol Trolox/g leaf d.w, respectively) than pomifera var. Conventional extraction showed lower amounts of phenolic compounds (7.81 ± 0.03 and 4.64 ± 0.01 mg/g leaf d.w. for flavonols and flavan-3ols, respectively) in comparison to the ultrasound-assisted ones.     

Enterohemorrhagic <Emphasis Type="Italic">Escherichia coli</Emphasis> (EHEC) in water from karst springs: detection with real-time PCR and isolation of strains

Within 2 months, two water sources in a karst area in Switzerland were sampled 9 times each, and analyzed by real-time PCR for 6 EHEC O-types, Shiga-like-toxin (stx1 and stx2) and intimin (eae) genes. With the exception of O111, 5 O-types were recorded regularly and at high frequencies (O26: 33.3 %; O157: 33.3 %; O104: 66.6 %; O103: 72.2 %; O145: 94.4 %). Genes for Shiga-like-toxins and intimin were almost omnipresent (stx1: 77.8 %; stx2: 83.3 %; eae: 77.8 %). Strain isolation was undertaken for O-groups 26, 103, 104, 145 and 157. Sample selection for strain isolation was based on Cq-values for the O-groups and stx1, stx2 and eae. From selected samples, frozen enrichment cultures were cultivated on EHLY-agar and 50 typical colonies screened for the O-type and genes encoding for stx1, stx2 and eae. With this approach, only one virulent EHEC-strain could be isolated (Escherichia coli O103, stx1 +; stx2 ?; eae +). We carried out one extensive testing with 800 colonies of O-group O145, and no virulent strain was isolated. Our findings showed that PCR-results are not sufficient to formulate epidemiological conclusions and that the isolation of strains is necessary. However, as the detection procedure of EHEC in foods is cumbersome and expensive, the appropriateness of such an approach in official food control is a matter of debate.     

Probiotic lactic acid bacteria isolated from traditional Korean fermented foods based on β-glucosidase activity

This study was designed to isolate lactic acid bacteria (LAB) with β-glucosidase activity and probiotic properties from Korean fermented foods. Among nine isolates, four LAB strains had excellent survival rates at pH 2.5 with 0.3% (w/v) pepsin for 3 h and 0.3% (w/v) oxgall for 24 h. Four LAB strains did not produce β-glucuronidase and showed adhesion ability to HT-29 cells that was superior to that shown by the reference strain Lactobacillus rhamnosus GG. All four strains were sensitive to ampicillin, tetracycline, chloramphenicol, and doxycycline. These strains were identified as Leuconostoc mesenteroides H40, Lactobacillus plantarum FI10604, L. brevis FI10700, and L. perolens FI10842 by 16S rRNA gene sequence, respectively. It was found that L. perolens FI10842 produced the highest β-glucosidase activity (49.10 mU/mL). These results indicate that the four LAB strains could be used as potential probiotic. Especially L. perolens FI10842 could be used as a starter culture for fermentation.     

Validation of a Loop-Mediated Amplification/ISO 6579-Based Method for Analysing Soya Meal for the Presence of <Emphasis Type="Italic">Salmonella enterica</Emphasis>

An alternative method for detection of Salmonella spp. in animal feed, based on the use of loop-mediated amplification (LAMP) in conjunction with a standard culturing procedure, was compared with the standard ISO 6579 as reference method, using soya meal as the test matrix. In the method comparison study, the sensitivities for both the alternative and reference methods were 100 %. The relative level of detection was 1.000. Tested against 100 Salmonella and 30 non-Salmonella strains, the LAMP-based method was 99 % inclusive and 100 % exclusive. The interlaboratory trial involved ten laboratories from eight European countries, testing eight samples at three contamination levels: 0 cfu/100 g, 1–5 cfu/100 g and 14–68 cfu/100 g. The trial specificity, or percentage correct identification of uncontaminated samples, was 96.3 % for both the reference methods and the LAMP/ISO 6579 alternative method, thus demonstrating its suitability for adoption as a procedure for rapid identification of Salmonella uncontaminated soya meal.     

Partition Behaviors of Different Polar Anthocyanins in Aqueous Two-Phase Systems and Extraction of Anthocyanins from <Emphasis Type="Italic">Nitraria tangutorun</Emphasis> Bobr. and <Emphasis Type="Italic">Lycium ruthenicum</Emphasis> Murr.

This study aimed to investigate the partition behaviors of various polar anthocyanins in NaH₂PO₄/(NH₄)₂SO₄-ethanol aqueous two-phase systems (ATPS) and to extract anthocyanins from Nitraria tangutorun Bobr. and Lycium ruthenicum Murr. Anthocyanins in Hibiscus sabdariffa L., Morus atropurpurea Roxb., N. tangutorun, and L. ruthenicum were profiled using HPLC-ESI-MS/MS and HPLC-DAD, and the partition behaviors of total anthocyanins and main anthocyanins were studied. The partition coefficient of anthocyanins increased with increased hydrophobicity, and low-polarity anthocyanins exhibited a higher preference for the top phase in NaH₂PO₄/(NH₄)₂SO₄-ethanol ATPS. Additionally, the NaH₂PO₄-ethanol ATPS gave higher selectivity and total anthocyanin yield than the (NH₄)₂SO₄-ethanol system. Extraction at 65 °C for 45 min and at 45.5 °C for 45 min using 28% NaH₂PO₄ and 26% ethanol (w/w) led to the recovery of 98.91 ± 0.03% of N. tangutorun anthocyanins (3.62 ± 0.05 mg/g) and 99.84 ± 0.01% of L. ruthenicum anthocyanins (13.16 ± 0.29 mg/g) from raw material; more than 70% of total sugars were removed in a single step. NaH₂PO₄-ethanol aqueous two-phase extraction is a promising method for extracting anthocyanins from N. tangutorun and L. ruthenicum.     

Grape Processing by High Hydrostatic Pressure: Effect on Use of Non-<Emphasis Type="Italic">Saccharomyces</Emphasis> in Must Fermentation

Vitis vinifera (variety Tempranillo) grapes were pressurized at 400 MPa by high hydrostatic pressure for 10 min and the effect on wild microbial populations, phenol extraction and wine composition was monitored. After treatment, the grapes were inoculated and fermented with Saccharomyces cerevisiae and several non-Saccharomyces yeasts: Schizosaccharomyces pombe, Torulaspora delbrueckii, Metschnikowia pulcherrima and Lachancea thermotolerans. S. pombe was used as sole fermentative yeast, but T. delbrueckii, M. pulcherrima and L. thermotolerans were used in sequential cultures with S. cerevisiae to completely ferment the sugars. The HHP treatment reduces strongly or eliminates wild microorganisms, especially yeasts, facilitating the growth and development of non-Saccharomyces yeasts. Thus, it helps to get either a better expression of enzymatic activities or metabolites production of non-Saccharomyces affecting wine quality.