Development of Real-time PCR Assays for the Detection of the <Emphasis Type="Italic">pin</Emphasis> II Terminator (t<Emphasis Type="Italic">pin</Emphasis>II) Used in GM Constructs and Its Donor Organism,Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis>)

Two singleplex TaqMan methods were developed for the detection of potato targets: one for the detection of the tpinII terminator, which is an emerging terminator used in GM constructs, and one for the detection of the endogenous StLS gene of potato. Performance criteria such as specificity and sensitivity were successfully tested for the two methods, taking into account the recommendations of international guidelines. The presence of the StLS target was checked in 16 potato cultivars. The StLS target is present at low copy number and can be used for quantitation purposes, as demonstrated on transgenic potatoes in this paper. The StLS target is an excellent candidate to replace the presently recommended endogenous target based on the UGP gene, which shows several disadvantages due to its high copy number and lack of specificity. The research also indicates that DNA can easily be extracted from different parts of potato tubers with a classical cetyltrimethylammonium bromide method.     

A First Approach of Using Ultrasound as an Alternative for Blanching in Vacuum-Packaged Potato Strips

The effect of ultrasound (US) (40 kHz, 200 W, 3 min), blanching (85 °C, 3.5 min), and the combination of both methods was evaluated on the quality of vacuum-packaged potato strips stored at 3 ± 1 °C for up to 10 days. For this study, two cultivars of potatoes were assessed. For blanched Agata samples, the lightness (L*) decreased over 12 % (p < 0.05). Moreover, their hue increased up to 100, obtaining lesser yellow potato strips. In contrast, US did not affect the hue values. The losses of firmness of blanched potato strips were notable (35 % for Agata and 51 % for Agria), whereas US did not change this property (p < 0.05). Nevertheless, no significant differences were found in the total starch content at 10 days. Agata and Agria showed different metabolic behaviors of sucrose in the refrigerated storage. Therefore, Agria cultivar retained better color after frying. These results suggest that US had less impact on color and improve the firmness in vacuum-packaged potato strips with no added chemicals.     

Modeling the Inactivation of <Emphasis Type="Italic">Listeria innocua</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> in Fresh-Cut Tomato Treated with Pulsed Light

The effectiveness of pulsed light (PL) treatments to inhibit microorganisms on fresh-cut tomatoes (Lycopersicon esculentum Mill., cv. Daniela) was investigated. Tomato slices inoculated with Escherichia coli or Listeria innocua were exposed to PL treatments (4, 6, or 8 J cm^?2 fluence) and kept cold at 4 °C for 20 days. L. innocua and E. coli counts, gases in the headspace of the containers (O₂ and CO₂), pH, titratable acidity, and soluble solid content were monitored throughout the cold storage. The PL treatments reduced significantly (p < 0.05) initial loads of both microbes. The effect of the PL fluence on the survival number of microoganisms was described by a log-linear model (R ² = 0.849–0.999). At any fixed time within the cold storing, the microbial counts for untreated samples were always higher than those cut tomatoes that had been previously PL-treated. The behavior of L. innocua and E. coli during the storage were well adjusted (R ² > 0.930) by Gompertzian models; the studied microorganisms exhibited different patterns during the storage period. On the other hand, O₂ and CO₂ partial pressures in containers with fresh-cut tomatoes were also significantly affected by PL treatments (p < 0.05). The highest PL fluence caused the greatest changes of O₂ and CO₂ contents. In addition, the application of PL triggered an acceleration of the O₂ consumption during the cold stage. PL treatments might be used to effectively extend the safety of fresh-cut tomatoes over 12 days of storage against E. coli and L. innocua growth.     

Effect of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> Bauer and <Emphasis Type="Italic">Bifidobacterium animalis</Emphasis> ssp. <Emphasis Type="Italic">lactis</Emphasis> BB12 on proteolytic changes in dry-cured loins

The effect of the potentially probiotic bacteria strain of Lactobacillus acidophilus Bauer and probiotic bacteria Bifidobacterium animalis ssp. lactis BB12 on proteolytic changes of proteins in dry-cured loins during fermentation and cold storage was studied. Results of the conducted tests demonstrated that the use of probiotic bacteria for the production of dry-cured meats impacts the generation of products of protein proteolysis with high antioxidant activity. The highest antioxidant activity of peptides after fermentation and cold storage was observed in the loin with the strain B. animalis ssp. lactis BB12 and the loin with the mixture of strains L. acidophilus Bauer and B. animalis ssp. lactis BB12. Qualitative analysis of peptides demonstrated that peptides with weight below 3.5 kDa are characterized by the highest capacity of quenching the ABTS cation radical, including the peptides in loins with the strain B. animalis ssp. lactis BB12.     

Nutritional,functional, thermal and structural characteristics of <Emphasis Type="Italic">Citrullus lanatus</Emphasis> and <Emphasis Type="Italic">Limonia acidissima</Emphasis> seed flours

Seeds from fruits such as Citrullus (C.) lanatus (watermelon) and Limonia (L.) acidissima (wood apple) are not commonly utilized but could be suitable in numerous food formulations. It was shown that the protein content of defatted seed flours was 71.38 and 49.51 % and that these contained considerable amounts of minerals such as Na, Mn, Mg, K, Cu, Fe and Zn. The defatted L. acidissima seed flour was superior to C. lanatus in essential amino acids. The flours obtained from both seeds were also evaluated for functional properties and characterized by X-ray diffraction, differential scanning calorimeter and scanning electron microscope (SEM). Amorphous nature was observed in defatted C. lanatus and L. acidissima flours due to the low percentage of degree of crystallinity. Spherical morphologies were observed through SEM. The exothermic peak was recorded in defatted C. lanatus and L. acidissima flour.     

Kinetics of Quality Changes of Shrimp (<Emphasis Type="Italic">Litopenaeus setiferus</Emphasis>) During Pasteurization

Effect of cooking between 1 and 80 min at 60 to 100 °C on several quality attributes of whole peeled shrimp (Litopenaeus setiferus) (80–90 counts/kg) was studied using an isothermal heating method. Cook loss, area shrinkage, and hardness of shrimp increased with increasing heating time and temperature, following a fractional first-order kinetic model with activation energies (E _a ) of 71.0, 53.3, and 29.9 kJ/mol, respectively. Cook loss, area shrinkage, and hardness were positively correlated. The toughness of shrimp muscle increased in the initial period of heating, then decreased in the later period during the treatments. The overall color change (ΔE) increased with increasing treatment time and temperature, and followed a zero kinetic model with an E _a of 37.2 kJ/mol. The kinetic parameters obtained from this study can be applied toward understanding and predicting shrimp-quality changes during pasteurization treatments, and further provides insight into the pasteurization conditions required to achieve safe and high-quality shrimp products and potentially other crustacean shellfish and seafood products.     

Effect of Low-pressure Cold Plasma (LPCP) on the Wettability and the Inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> on Fresh-Cut Apple (<Emphasis Type="Italic">Granny Smith</Emphasis>) Skin

The aim of this study was to investigate the effect of low-pressure cold plasma (LPCP) on the inactivation of Escherichia coli and Listeria innocua on fresh-cut apple skin and its influence on wettability. Cold plasma treatments have shown to be effective to decontaminate foods, but their effect on the wettability has not been well studied. Surface-inoculated apple samples were treated with argon (Ar), nitrogen (N2), oxygen (O₂), and argon-oxygen (Ar-O₂) cold plasma using a commercial LPCP unit. Three different models were used to fit bacterial survival curves after the LPCP treatments. Changes in surface wettability were also determined by measuring the contact angle. The LPCP treatments using Ar, O₂, or Ar-O₂ mixture for 20 min were the most effective to inactivate E. coli with O₂, while the LPCP treatment with N2 for 20 min reduced L. innocua the most for (p?<?0.05). The highest increase in surface wettability was observed in samples treated for 20 min with O₂ and Ar-O₂. Different LPCP treatments have not great effectivity on the inactivation of E. coli and L. innocua on fresh-cut apple surface, but the all treatments changed the surface wettability of apples, making it more hydrophilic. This can be considered as a negative effect of the LPCP treatment because it can facilitate the adhesion and proliferation of re-contaminating microorganisms. More research should be undertaken to explore the use of other gases and complex surfaces.     

Effect of heat-treated <Emphasis Type="Italic">Nuruk</Emphasis> on the quality characteristics of aged <Emphasis Type="Italic">Yakju</Emphasis>

Long-term aging of Yakju, a traditional Korean liquor made of rice and Nuruk (a fermentation agent), causes browning and odor and flavor development. This study investigated the effects of heat-treated Nuruk (50–80 °C, 30 min) on Yakju quality. The saccharogenic powers and glucoamylase, α-amylase, and carboxypeptidase activities were similar in non-heat-treated Nuruk and that treated at 50 °C. However, acidic protease and alcohol dehydrogenase decreased above 50 °C. The content of nitrogen-containing compounds was inversely proportional to the heat-treatment temperature. Compounds that cause off-flavors decreased at 50–60 °C, but increased at 70–80 °C, whereas compounds that provide fragrance increased at 50–60 °C. Sensory evaluation indicated that bad taste attributes were higher in Yakju produced using non-heat-treated Nuruk. Therefore, heat treatment of Nuruk at 50 °C can be adopted as a method for improving Yakju quality, as enzymatic activities that affect color, aroma, and taste are regulated.     

Probe-Based Fluorescence Melting Curve Analysis for Differentiating <Emphasis Type="Italic">Larimichthys polyactis</Emphasis> and <Emphasis Type="Italic">Larimichthys crocea</Emphasis>

Larimichthys polyactis (redlip yellow croaker) and Larimichthys crocea (large yellow croaker) are commercially important fish species in East Asia with high differences in their market values. In Korea, consumers prefer L. polyactis to L. crocea, although it is difficult to distinguish them based on their morphological traits. The objective of this study was to develop an assay for differentiating L. polyactis and L. crocea using fluorescence melting curve analysis (FMCA) with a single locked nucleic acid (LNA) probe. Species-specific regions of the mitochondrial 16S rDNA were selected as LNA probes. The target sequences of L. polyactis and L. crocea had a 2-bp difference, and a single LNA probe was identified using melting temperature (Tm) shift. LNA probe was 100 % complementary to the target sequence of ten L. polyactis samples, giving a significantly higher Tm value (66 °C) than that of five L. crocea samples (42 °C). Overall, the developed LNA-based FMCA system had high efficiency, multiplexity, and simplicity, and could be effectively used for differentiating L. polyactis and L. crocea, and as a food analyzing method based on DNA sequence.     

Differences of Bactericidal Efficacy on <Emphasis Type="Italic">Escherichia coli</Emphasis>, <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>, and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> of Slightly and Strongly Acidic Electrolyzed Water

In the present study, the disinfection efficacy of slightly acidic electrolyzed water (SAEW) and strongly acidic electrolyzed water (AEW) was tested on three bacteria, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis and the disinfection mechanism was discussed. The results showed that SAEW had a stronger antibacterial efficacy against these tested bacteria in comparison with AEW. The results also showed that both SAEW and AEW treatments could damage the cell membrane, which was demonstrated microcosmically by scanning electron microscopy (SEM), thus causing leakages of protein, DNA, RNA, and ATP, resulting in the death of microbes. Moreover, AEW treatment could not cause the degradations of DNA and RNA, and nucleic acids including DNA and RNA are not the target point of its bactericidal efficacy. However, SAEW could maybe cause the degradation of RNA, and RNA may be the target in its antibacterial activity. We suggested that the differences in antibacterial efficacy between SAEW and AEW could be explained by the different impacts on RNA of tested strains.     

Characterization of brewing microorganisms isolated from Korean traditional <Emphasis Type="Italic">nuruk</Emphasis> for <Emphasis Type="Italic">Cheongju</Emphasis> production

The objective of this study was to select and identify yeasts and molds isolated from traditional nuruk and to investigate their brewing characteristics for Cheongju production. The yeast strains Y190 (Accession ID-KACC 93251P), Y263 (Accession ID-KACC 93252P), and Y270 (Accession ID-KACC 93253P) showing high alcohol and flavor productivity were isolated and identified by phylogenetic inference based on an internal transcribed spacer 2 region sequence analysis. In addition, Aspergillus oryzae 83-10 (Accession ID-KACC 93254P) showing the highest enzyme activity was isolated. This study provides basic data for the production of Korean Cheongju and assesses the applicability of these three yeast strains and A. oryzae 83-10 isolated from traditional nuruk.     

Effect of different cooking methods on vitamins,minerals and nutritional quality indices of orange-spotted grouper (<Emphasis Type="Italic">Epinephelus coioides</Emphasis>)

This study aimed to evaluate the nutritional value [proximate, fatty acid profiles, vitamins and minerals and also nutritional quality indices (NQI)] of orange-spotted grouper (Epinephelus coioides) prepared according to common consumer techniques: raw, poached, steamed, microwaved, pan-fried and deep-fried (in olive oil). There was an increase in protein, lipid and ash contents in cooked fish as compared to raw fish fillets. Cooking treatment had no significant effect on total n-3 fatty acids except for deep-fried fillets. Cooking methods had no significant effect on total n-6 fatty acids. In all the cooking methods, n-6 fatty acids were well preserved. There was no significant difference in the Mn, Zn, Ca and Fe contents of fillet between raw and cooked samples. Vitamin D content in all cooking methods did not differ significantly. None of cooking methods had a significant effect on vitamin A except for deep-frying fillets. However, vitamin B1 and B3 contents of cooked fish significantly decreased. Considering the overall nutritional quality indices, vitamin and mineral contents, steaming is the best cooking method.     

Exposure assessment of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> in <Emphasis Type="Italic">Kimbab</Emphasis>, a ready-to-eat Korean food

The exposure assessment was carried out for Staphylococcus aureus in kimbab by predicting growth of S. aureus and the production of enterotoxin using Food MicroModel® program. Environmental parameters selected were pH 5.5, Aw 0.999, and storage temperatures in the range of 10 to 30°C. It was predicted that 6.3 hr could be a critical time for enterotoxin production while kimbab was stored at 30°C. Mild case scenario analysis showed that enterotoxin could not be produced if kimbab was kept at 10°C during preparation and distribution and then left at 25°C for 4 hr before consumption. In the worst case scenario, the keeping time at 25°C was assumed to be 7.0 hr. The level of S. aureus in the worst case was predicted to be 6.8×10⁶ CFU/g which is lower than the critical level (7.8×10⁶ CFU/g) for toxin production.     

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.     

Synergistic interaction of <Emphasis Type="Italic">Auricularia auricula</Emphasis>-<Emphasis Type="Italic">judae</Emphasis> polysaccharide with yam starch: effects on physicochemical properties and in vitro starch digestibility

Thermal stable polysaccharides from Auricularia auricula-judae (AP) have unique molecularstructures and multiple bioactivities. The effects of AP on the physicochemical properties and in vitro starch digestibility of yam starch (YS) were studied. The addition of AP induced a significant increase in the swelling power, solubility, mean volume diameter and adhesiveness as well as a dramatic decrease in the hardness and gumminess (p?<?0.05). AP showed a strong suppressive effect on in vitro starch digestibility. Higher modulus (G′, G″) and stiffness parameters (A_α), and lower order of relaxation function (α), were observed in oscillatory rheological measurements, indicating that the gels were more elastic-like and had higher pseudoplasticity in the presence of AP. Furthermore, AP remarkably decreased the syneresis and storage modulus (G′), and also retarded the retrogradation process of YS gel at 4°C, revealing a synergistic interaction between AP and YS, which could also be demonstrated by scanning electron microscopy.     

Integrated Analytical Methods to Characterize Lipids from <Emphasis Type="Italic">Prosopis</Emphasis> spp. and <Emphasis Type="Italic">Ceratonia siliqua</Emphasis> Seed Germ Flour

Flour from seed germ of European carob (Ceratonia siliqua) and South American algarrobo (Prosopis spp.) is a potential ingredient for health-promoting baked products. Herein, lipids from germ of three Argentinean Prosopis (P. alba, P. nigra, and P. ruscifolia) and one European carob species were characterized in detail, exploiting an array of up-to-date analytical techniques. Total lipids ranged from 7.1 to 8.1% (w/w). Linoleic acid (C18:2, ω-6) predominated the GC flame ionization detector profiles of fatty acid in all samples (43.3–50.6%). Prosopis spp. contained 6–7% of C20–C24 fatty acids and, consistently, C56–C60 triacylglycerols, as detected by MALDI-TOF mass spectrometry (MS), which were practically missing in C. siliqua germ. Phospholipids were isolated by hydroxyapatite chromatography, characterized by MALDI-TOF MS and grossly quantified by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. Because of a relatively high content (9.5–11.8%, v/v), phospholipids might increase the antioxidant potential and improve the baking performances of flour fortified with carob and algarrobo seed germ.     

Comparative Study on the Inactivation and Photoreactivation Response of <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> Seafood Isolates and a <Emphasis Type="Italic">Listeria innocua</Emphasis> Surrogate after Pulsed Light Treatment

The inactivation and photoreactivation response of six seafood-isolated Listeria monocytogenes and one Listeria innocua strain after pulsed light (PL) treatment was evaluated. The lower inactivation levels found after exposure of treated samples to daylight during the first 90 min of storage confirmed that both L. innocua and L. monocytogenes have the capability to photorepair PL-induced DNA damage upon appropriate conditions. Photoreactivation levels from 0.2 to 2.1 log CFU cm^?2 were observed depending on treatment intensity (fluence) and Listeria strain. Complete photorepair of PL-caused damage was not found even after treatments inducing low inactivation levels. Photoreactivation increased up to 2.1 log with the applied fluence up to a threshold able to cause between 2.4 and 5.4 log reductions under dark storage. Photorepair was not avoided but lower photoreactivation was observed after higher fluence inducing more than 6 log reductions under dark storage. Both L. innocua and L. monocytogenes serotype 1/2b exhibited the highest photoreactivation levels whereas serotypes 1/2a showed the lowest ones. The overall inactivation and photoreactivation responses of tested Listeria strains were comparable indicating that L. innocua may be a good surrogate for the safe evaluation, optimization and validation of PL technology to control L. monocytogenes in food products and food processing facilities.     

Effect of germinated grain flours on physicochemical characteristics of rice cakes, <Emphasis Type="Italic">Seolgitteok</Emphasis>

Minor grains and their germinated shoots such as buckwheat, Job's tears, and mungbean have diverse bioactive functionalities. In this study, the flours of grains and germinated grains were added to Korean rice cakes, known as Seolgitteok, and the effects on properties such as starch gelatinization and retrogradation were examined. The dietary fiber content, blue value index, amylogram, texture properties and sensory evaluation of Seolgitteok were also investigated. Rice flour amylogram results showed that addition of buckwheat, germinated Job’s tears, and mungbean decreased starch setback, and that addition of buckwheat decreased consistency, which are related to slowed retrogradation. Buckwheat, especially when germinated, delayed starch retrogradation, which is one of the main drawbacks of rice cakes. Seolgitteok with added germinated buckwheat possessed preferable sensory qualities.     

Effect of <Emphasis Type="Italic">Punica granatum</Emphasis> peel and <Emphasis Type="Italic">Melia azedarach</Emphasis> bark extracts on durability of European beech and maritime pine

A method to improve wood durability using natural extracts was evaluated. Wood deterioration is a condition caused by several abiotic and biotic factors including fungal contamination. To date, approaches aiming at the reduction of these contaminants mainly involve the use of chemicals agents. Natural products could represent an alternative strategy. Aqueous extracts of Punica granatum L. (pomegranate) peel and Melia azedarach L. barks were evaluated as antifungal agents to improve natural durability of beech wood and maritime pine. To evaluate the effect of treatments under simulated accelerated ageing of wood by natural conditions, impregnation and leaching tests were performed. Results demonstrated that samples impregnated with pomegranate or M. azedarach solutions notably increased the biological resistance of wood in a dose-dependent manner. These results were confirmed by the reduction in weight losses in treated samples even after 6 weeks of fungal exposure. Moreover, after leaching tests, 20 and 7% (w/v) of pomegranate and M. azedarach extract solutions were demonstrated as the better concentrations to enhance wood durability. Total phenol content and characterization of the phenolic compounds in both, natural extracts and wood samples were analyzed by Folin–Ciocalteu assay and HPLC-DAD. In conclusion, it was demonstrated that the present method can be considered as an effective treatment to increase wood durability while it proposes the valorization of natural extractives in wood industry.