Combined Effect of Citric Acid and Polyphenol-Rich Grape Seed Extract towards Bioactive Smart Food Packaging Systems.

Alginate films (2% w·v-1) were prepared with varying concentrations (5-20% w/w) of citric acid and aqueous grape seed extract (GSE) filtrate (11.66 ± 1.32 g GAE/L) using the solvent-evaporation method. Crosslinking alginate via ester bonds (FTIR analysis) with citric acid up to 10% (w/w) led to a 33% increase in tensile strength, a 34% reduction in water vapor transmission rate (WVTR), and had no impact on elongation at break. Crosslinking alginate with citric acid in the presence of GSE increased the tensile strength by 17%, decreased WVTR by 21%, and significantly improved DPPH scavenging activity. Moreover, after incubation for 24 h at 37 °C, the film-forming solutions exhibited increased antimicrobial activity, resulting in 0.5- and 2.5-log reductions for Escherichia coli and Staphylococcus aureus, respectively, compared to the values obtained without the addition of GSE. The stronger inhibitory effect observed against Gram-positive bacteria can be attributed to the unique composition and structure of their cell walls, which creates a barrier that restricts the penetration of polyphenols into the cells. The pH adjustment of the GSE film-forming solution from 2.0 to 10.0 shifted the UV/VIS absorption spectra, resulting in a colour change from yellow to red. The findings of this study have showcased the potential of combining GSE and citric acid to enhance the functionality and bioactivity of alginate films for applications in smart food packaging.

Susceptibility and transcriptomic response to plasma-activated water of Listeria monocytogenes planktonic and sessile cells.

Plasma-Activated Water (PAW) was generated from tap water using a surface dielectric barrier discharge at different discharge power (26 and 36 W) and activation time (5 and 30 min). The inactivation of a three-strain Listeria monocytogenes cocktail in planktonic and biofilm state was evaluated. PAW generated at 36 W-30 min showed the lowest pH and the highest hydrogen peroxide, nitrates, nitrites contents and effectiveness against cells on planktonic state, resulting in 4.6 log reductions after a 15-min treatment. Although the antimicrobial activity in biofilms formed on stainless steel and on polystyrene was lower, increasing the exposure time to 30 min allowed an inactivation >4.5 log cycles. The mechanisms of action of PAW were investigated using chemical solutions that mimic its physico-chemical characteristics and also RNA-seq analysis. The main transcriptomic changes affected carbon metabolism, virulence and general stress response genes, with several overexpressed genes belonging to the cobalamin-dependent gene cluster.

Mechanical, Barrier, Antioxidant and Antimicrobial Properties of Alginate Films: Effect of Seaweed Powder and Plasma-Activated Water.

The incorporation of natural fillers such as seaweed may potentially enhance the properties of biopolymer films. In this study, we investigated the effect of seaweed powder as a bio-filler in alginate-based films at different concentrations (10, 30, and 50%, w/w alginate) and particle sizes (100 and 200 µm) on the mechanical, barrier, antioxidant, and antimicrobial properties of alginate which are essential for food packaging applications. Initially, mechanical properties of the alginate films prepared at different temperatures were evaluated to find the optimal temperature for preparing alginate solution. The addition of seaweed powder did not have any positive effect on the mechanical properties of the alginate films. However, the barrier (water vapor transmission rate) and antioxidant properties were improved with the addition of seaweed filler regardless of concentration. In addition, selected films were prepared in plasma-activated water (PAW). The mechanical properties (tensile strength, but not elongation at break) of the films prepared with PAW improved compared to the films prepared in distilled water, while a significant decrease was observed when incorporated with the seaweed filler. The films prepared in PAW also showed improved barrier properties compared to those prepared in distilled water. The antimicrobial activity of the alginate-seaweed film-forming solution was in general more pronounced when prepared with PAW and stored at 10 °C, particularly at the highest concentration of the film-forming solution (83.3% v/v). A more pronounced inhibitory effect was observed on the Gram-positive S. aureus than on the Gram-negative E. coli, which has been attributed to the different composition and structure of the respective cell walls. This study has demonstrated the potential of seaweed filler in combination with PAW towards enhanced functionality and bioactivity of alginate films for potential food packaging applications.

Effect of Citric Acid Cross Linking on the Mechanical, Rheological and Barrier Properties of Chitosan.

In this study, acetic acid (AA-2% w/v), a combination of acetic acid and citric acid (AA-1% w/v + CA-1% w/w), and three different concentrations of citric acid (CA-2, 4 and 6% w/w) were used to create chitosan solution. The FTIR analysis showed the presence of residual CA in all the CA-containing samples where no trace of AA was observed. The tensile strengths of the CA-containing samples were lower than the AA samples. Whereas the values for the elongation at break of the CA samples were higher than the AA samples, which kept increasing with an increasing CA content due to the plasticizing effect from residual citric acid. The elongation at break values for 4 and 6% CA-containing samples were 98% higher than the AA samples. The samples prepared with CA showed shorter LVE regions that reduced with an increasing CA concentration compared to the AA samples. Different acid concentrations did not have a large effect on the gelation time. However, CA-containing samples showed higher viscosities as compared to the AA-containing solution, which increased with an increasing CA content. The water vapour transmission rates of the CA-containing samples were lower than the others. All the chitosan solutions suppressed the growth of the two test strains, and none of the variants reached an abs 600 nm at 0.2.

Impact of Plasma-Activated Water Treatment on Quality and Shelf-Life of Fresh Spinach Leaves Evaluated by Comprehensive Metabolomic Analysis.

Fresh baby spinach leaves are popular in salads and are sold as chilled and plastic-packed products. They are of high nutritional value but very perishable due to microbial contamination and enzymatic browning resulting from leaf senescence. Therefore, innovative food processing methods such as plasma-activated water (PAW) treatment are being explored regarding their applicability for ensuring food safety. PAW's impact on food quality and shelf-life extension has, however, not been investigated extensively in vegetables so far. In the present study, a comprehensive metabolomic analysis was performed to determine possible changes in the metabolite contents of spinach leaves stored in a refrigerated state for eight days. Liquid chromatography high-resolution mass spectrometry, followed by stringent biostatistics, was used to compare the metabolomes in control, tap-water-rinsed or PAW-rinsed samples. No significant differences were discernible between the treatment groups at the beginning or end of the storage period. The observed loss of nutrients and activation of catabolic pathways were characteristic of a transition into the senescent state. Nonetheless, the presence of several polyphenolic antioxidants and γ-linolenic acid in the PAW-treated leaves indicated a significant increase in stress resistance and health-promoting antioxidant capacity in the sample. Furthermore, the enhancement of carbohydrate-related metabolisms indicated a delay in the senescence development. These findings demonstrated the potential of PAW to benefit food quality and the shelf-life of fresh spinach leaves.

Synthesis of Sodium Alginate-Silver Nanocomposites Using Plasma Activated Water and Cold Atmospheric Plasma Treatment.

In this study, sodium alginate (SA)-based, eco-friendly nanocomposites films were synthesized for potential food packaging applications using silver nitrate (AgNO3) as the metal precursor, reactive nitrogen and oxygen species (RNOS) created within plasma activated water (PAW), or through cold plasma treatment (CP) as reducing agent and SA as stabilizing agent. The formation of silver nanoparticles (AgNPs) was confirmed via the absorption peaks observed between 440 and 450 nm in UV-vis spectroscopy. The tensile strength (TS) and tensile modulus (TM) of the nanocomposite films were significantly higher than those of the SA films. An increase in the TS was also observed as the AgNP concentration was increased from 1 to 5 mM. The storage modulus (G') of the nanocomposite solution was higher than that of the SA solution. The synthesis of AgNPs resulted both in a higher solution viscosity and a more marked shear-thinning effect. The synthesized AgNPs showed antimicrobial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. The AgNPs were spherical in shape with an average size of 22 nm.

PLA-Based Materials Containing Bio-Plasticizers and Chitosan Modified with Rosehip Seed Oil for Ecological Packaging.

Several recipes based on PLA, bio-plasticizers, and active agents such as vitamin E and cold-pressed rosehip seed oil encapsulated into chitosan by the emulsion method named here as chitosan modified (CS-M) were elaborated by melt compounding for food packaging applications. Resulted biocomposites have been investigated from the point of view of physical-mechanical, thermal, barrier, antimicrobial, and antioxidant properties to select the formulations with the optimum features to produce food trays and films for packaging applications. The obtained results showed that the elaborated formulations exhibit tensile strength and flexibility dependent on their composition being either rigid or flexible, as well as antimicrobial and antioxidant activity, which will potentially lead to prolonged use for food packaging. The recipe with PLA matrix and 40:60 Lapol®108 as masterbarch/polyethylene glycol (MB/PEG) bio-plasticizers ratio was distinguished by an improvement of over 100 times in terms of flexibility compared with neat PLA, while the highest antioxidant activity (36.27%) was recorded for the sample containing a CS-M and MB/PEG ratio of 60:40. An enhancement of ~50% for the water vapor barrier was recorded for PLA/CS-M_100:0 material. By modulating the MB and PEG bio-plasticizers ratio, the design of new eco-friendly food packaging materials with antimicrobial/antioxidant characteristics by using the existing technologies for processing synthetic polymers (melt mixing, compounding, pressing, thermoforming) has been successfully realized.

Innovative Ultrasound-Assisted Approaches towards Reduction of Heavy Metals and Iodine in Macroalgal Biomass.

The aim of this work was to evaluate the potential of ultrasound (US), alone or in combination with mild heating and/or EDTA towards reduction of As, Cd, I, and Hg content of Laminaria hyperborea. Concentrations of As, Cd, I, and Hg of 56.29, 0.596, 7340, and <0.01 mg kg-1 of dry weight, respectively, were found in L. hyperborea blades. Treatment with US at 50 °C increased approx. 2-fold the amount of As released, although did not affect significantly the content of Cd or I, as compared to control (no US) samples. Reducing the temperature to 8 °C significantly decreased the effect of US, but heating at 80 °C did not cause a significant effect as compared to treatments at 50 °C. On the other hand, treatment with 0.1 N EDTA at 50 °C enhanced the percentage of Cd released by approximately 7-fold, regardless of sonication. In the present work, the combination of US and EDTA at 50 °C for 5 min led to a significant reduction of the As (32%), Cd (52%) and I (31%) content in L. hyperborea, thus improving the product's safety for consumers.

Current status of biobased and biodegradable food packaging materials: Impact on food quality and effect of innovative processing technologies.

Fossil-based plastic materials are an integral part of modern life. In food packaging, plastics have a highly important function in preserving food quality and safety, ensuring adequate shelf life, and thereby contributing to limiting food waste. Meanwhile, the global stream of plastics into the oceans is increasing exponentially, triggering worldwide concerns for the environment. There is an urgent need to reduce the environmental impacts of packaging waste, a matter raising increasing consumer awareness. Shifting part of the focus toward packaging materials from renewable resources is one promising strategy. This review provides an overview of the status and future of biobased and biodegradable films used for food packaging applications, highlighting the effects on food shelf life and quality. Potentials, limitations, and promising modifications of selected synthetic biopolymers; polylactic acid, polybutylene succinate, and polyhydroxyalkanoate; and natural biopolymers such as cellulose, starch, chitosan, alginate, gelatine, whey, and soy protein are discussed. Further, this review provides insight into the connection between biobased packaging materials and innovative technologies such as high pressure, cold plasma, microwave, ultrasound, and ultraviolet light. The potential for utilizing such technologies to improve biomaterial barrier and mechanical properties as well as to aid in improving overall shelf life for the packaging system by in-pack processing is elaborated on.

Towards the Next-Generation Disinfectant: Composition, Storability and Preservation Potential of Plasma Activated Water on Baby Spinach Leaves.

Plasma activated water (PAW) has rapidly emerged as a promising alternative to traditional sanitizers applied in the fresh produce industry. In the present study, PAW chemistry and storage stability were assessed as a function of plasma operating conditions. Increasing plasma exposure time (5, 12.5, 20 min) and power (16, 26, 36 W) led to a significant drop in pH (2.4) and higher nitrates and nitrites levels (320 and 7.2 mg/L, respectively) in the PAW. Non-detectable hydrogen peroxide concentration, irrespective of the treatment conditions, was attributed to its instability in acidic environments and the remote PAW generation mode. pH, nitrates and nitrites levels in the PAW remained unaffected after two weeks at 4 °C. The potential of PAW for microbial inactivation and quality retention was demonstrated on baby spinach leaves. Rinsing steps influenced colour development during chilled storage to a greater extent than PAW treatment itself. About 1 log reduction in total bacterial counts (5 log CFU/g) was achieved through PAW rinsing, with no variability after eight days at 4 °C (typical shelf-life at retailers). Moreover, microbial levels on PAW-treated samples after storage were significantly lower than those on control samples, thus contributing to extended product shelf-life and reduced food waste generation.

Antioxidant/Antibacterial Electrospun Nanocoatings Applied onto PLA Films.

Polylactic acid (PLA) films were coated by coaxial electrospinning with essential and vegetable oils (clove and argan oils) and encapsulated into chitosan, in order to combine the biodegradability and mechanical properties of PLA substrates with the antimicrobial and antioxidant properties of the chitosan⁻oil nanocoatings. It has been established that the morphology of the electrospun nanocoatings mainly depend on the average molecular weight (MW) of chitosan. Oil beads, encapsulated into the main chitosan nanofibers, were obtained using high-MW chitosan (Chit-H). Oil encapsulated in chitosan naoparticles resulted when low-MW chitosan (Chit-L) was used. The coating layer, with a thickness of 100 ± 20 nm, had greater roughness for the samples containing Chit-H compared with the samples containing Chit-L. The coated PLA films had higher antibacterial activity when the nanocoating contained clove oil rather than when argan oil was used, for both types of chitosan. Nanocoatings containing Chit-H had higher antibacterial activity compared with those containing Chit-L, for both types of oil tested, due to the larger surface area of the rougher nanoscaled morphology of the coating layer that contained Chit-L. The chitosan⁻clove oil combination had higher antioxidant activity compared to the simple chitosan nanocoating, which confirmed their synergistic activities. The low activity of systems containing argan oil was explained by big differences between their chemical composition and viscosity.

Comparative Analysis of the Composition and Active Property Evaluation of Certain Essential Oils to Assess their Potential Applications in Active Food Packaging.

The antifungal, antibacterial, and antioxidant activity of four commercial essential oils (EOs) (thyme, clove, rosemary, and tea tree) from Romanian production were studied in order to assess them as bioactive compounds for active food packaging applications. The chemical composition of the oils was determined with the Folin-Ciocâlteu method and gas chromatography coupled with mass spectrometry and flame ionization detectors, and it was found that they respect the AFNOR/ISO standard limits. The EOs were tested against three food spoilage fungi-Fusarium graminearum, Penicillium corylophilum, and Aspergillus brasiliensis-and three potential pathogenic food bacteria-Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes-using the disc diffusion method. It was found that the EOs of thyme, clove, and tea tree can be used as antimicrobial agents against the tested fungi and bacteria, thyme having the highest inhibitory effect. Concerning antioxidant activity determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid (ABTS) methods, it has been established that the clove oil exhibits the highest activity because of its high phenolic content. Promising results were obtained by their incorporation into chitosan emulsions and films, which show potential for food packaging. Therefore, these essential oils could be suitable alternatives to chemical additives, satisfying the consumer demand for naturally preserved food products ensuring its safety.

Detection of small bioactive peptides from Atlantic herring (Clupea harengus L.).

Recent research has shown that fish residual materials contain a range of components with interesting biological activity. Therefore, there is a great potential in the marine bioprocess industry to utilize these by-products as starting material for generating more valuable products. The aim of the present study was to search for bioactive peptides (in particular small natural bioactive peptides with molecular weight lower than 10 kDa) in Atlantic herring (Clupea harengus L.) by-products such as skin and more general residual materials. By such means a range of peptides with claimed interesting biological activities was found. Herein the activity of the detected bioactive peptides and strategies for isolating peptide fragments containing the bioactive motif is discussed. Identification of bioactive peptides in crude peptide/protein sources (skin and residual materials) was performed directly using a combination of mass spectrometry (Orbitrap), bioinformatics and database search. This method was a good angle of approach in order to map the potential in new species and species that have been very little studied.

Cod (Gadus morhua) muscle proteome cataloging using 1D-PAGE protein separation, nano-liquid chromatography peptide fractionation, and linear trap quadrupole (LTQ) mass spectrometry.

Because Atlantic cod (Gadus morhua) has high economic value and its protein-rich muscle tissue is a food source, an increased understanding of the effects and consequences of environmental, nutritional, biological, and industrial factors on meat quality is necessary. To gain insight into cod muscle tissue protein composition, a large-scale proteomics approach has been used. One-dimensional polyacrylamide gel electrophoresis, nanoflow liquid chromatography peptide separation, and linear trap quadrupole mass spectrometry were used to identify 4804 peptides, which retrieved 9113 cod expressed sequence tags (ESTs), which in turn were mapped to 446 unique proteins. The same data set identified 3924 proteins from the zebrafish protein database, which highlights the complementary value of the two approaches. The generated data sets will act as a foundation for studies related to physiological status assessment of cod under different environmental conditions, screening for diseases, and biomarker identification for assessment of fish quality during industrial processing and preservation.

Microflora assessments using PCR-denaturing gradient gel electrophoresis of ozone-treated and modified-atmosphere-packaged farmed cod fillets.

Denaturing gradient gel electrophoresis (DGGE) of a PCR-amplified 16S rDNA sequence was used to characterize changes in the microbial flora caused by ozone (O3) treatment of farmed cod (Gadus morhua). Portions of cod were produced under controlled conditions, bathed in fresh water supplemented with 2 ppm of O3 for 30 min, and packaged in modified atmosphere (MA: 60% CO2 and 40% N2) before 4 degrees C storage. Control samples were packaged in MA or air, without prior O3 treatment. Samples were analyzed by PCR-DGGE to determine the predominant bacterial flora and to examine possible differences in the microbial community due to O3 treatment. The DGGE analysis during the storage period showed that the O3 treatment produced no significant difference in the microbial flora compared with the controls. Sequencing of 16S rDNA detected the specific spoilage bacteria Photobacterium phosphoreum, Pseudomonas spp., Shewanella baltica, and Shewanella putrefaciens as the predominant bacteria in all samples. PCR-DGGE results were supported by culture and sensory analyses used in predicting product shelf life. Aerobic plate count, H2S-producing bacteria, and psychrotrophic bacterial counts demonstrated no significant extension of the shelf life of MA-packaged, O3-treated cod fillets.

Characterisation of the bacterial flora of modified atmosphere packaged farmed Atlantic cod (Gadus morhua) by PCR-DGGE of conserved 16S rRNA gene regions.

The present article describes the use of broad-range molecular analyses to characterise the microbial population of farmed Atlantic cod (Gadus morhua) packaged for the retail market. Cod was filleted post rigor, packaged in air or in modified atmosphere (MA) (50% CO(2):50% N(2) or 50% CO(2):50% O(2)) and stored at 0 degrees C for 11 days. To determine the community profiles of the samples the variable V3-region of the bacterial 16S rRNA gene were amplified by PCR, before the PCR products were separated by denaturing gradient gel electrophoresis (DGGE). From sequence analyses Pseudomonas spp. were found to be the predominant bacteria in oxygen enriched atmospheres, whereas the spoilage bacteria Photobacterium sp., Shewanella putrefaciens and Pseudomonas spp. dominated in CO(2):N(2) and air packaged samples. Additional microbial analyses by cultivation methods observed highest bacterial numbers in air stored samples, and both MA mixtures gave growth inhibition when measuring aerobic plate count, psychrotrophic bacteria and H(2)S-producing bacteria. The results show that PCR-DGGE can be applied to examine bacterial diversity and population shifts among different MA-packaged products.

Characterisation of the dominant bacterial population in modified atmosphere packaged farmed halibut (Hippoglossus hippoglossus) based on 16S rDNA-DGGE.

It is not well understood why Atlantic halibut (Hippoglossus hippoglossus) has longer shelf-life than most other white fish species. Our approach was to examine the microbiological diversity of the spoilage microbiota during modified atmosphere (MA) packaging of farmed Atlantic halibut. Portions were packaged with gas mixtures of CO(2):N(2) and CO(2):O(2) (50%:50%) and with air as a reference. The packages were stored at 4 degrees C and samples were taken 6 times during the 23 days of storage. Analyses with molecular techniques (PCR-DGGE) determined profiles of the bacterial populations in the various samples and sequencing detected the bacterial species present. In addition, samples were analysed for microbial, chemical and sensory parameters. The shelf-life was 10-13 days when stored in air and between 13 and 20 days for MA packages, with oxygen-enriched packages suggested as the better gas mixture, based on microbial growth and sensory scores. From sequence analyses of the bacterial population Photobacterium phosphoreum and Pseudomonas spp. were found to dominate in the halibut. Brochothrix thermosphacta was found in most samples at the end of the storage period. Shewanella putrefaciens was found sporadically and in low concentrations based on microbial methods, but not detected by PCR-DGGE.