Nanoliposomal delivery systems of natural antibacterial compounds; properties, applications, and recent advances.

Todays, nanoliposomes (NLPs) are considered as one of the most efficient nanocarriers to deal with bacteria, practically in food products. These nanodelivery systems are able to be loaded with different bioactive compounds. The main aim of this review is investigating recent approaches (mostly from the years of 2018 to 2022) regarding development of nanoliposomal natural antibacterial compounds. In this regard, NLPs alone, combined with films, coatings, or fibers, and in coated forms are reviewed as advanced delivery systems of antibacterial substances. Moreover, a robust and comprehensive coverage of the morphological and physical properties of formulated NLPs as well as their interactions with antibacterial substances are discussed. The importance of NLPs to encapsulate antibacterial ingredients, advantages and drawbacks, antibacterial pathways of formulated NLPs, and comparison of them with pure antibacterial bioactive compounds are also explained.

Valorization of Amaranth (Amaranthus cruentus) Grain Extracts for the Development of Alginate-Based Active Films.

This research work investigates the development of alginate-based films incorporating phenolic compounds extracted from Amaranthus cruentus grain using different solvents. Alginate, glycerol, and amaranth grain phenolic compounds at various concentrations were used to produce the films. An experimental Central Composite Rotatable Design (CCRD) was used to evaluate the effect of these variables on different film's properties, i.e., water vapor permeability, hydrophobicity, moisture content, solubility, thermal, mechanical, and optical properties. This study demonstrated that high phenolic compound content and antioxidant capacity were obtained from amaranth grain using ethanol as the extraction solvent. Alginate films incorporating amaranth phenolic compounds were successfully manufactured, and this study can be used to tailor the formulation of alginate films containing amaranth phenolic compounds, depending on their final food application. For example, less flexible but more resistant and water-soluble films can be produced by increasing the alginate concentration, which was confirmed by a Principal Component Analysis (PCA) and Partial Least Squares (PLS) analysis. This study showed that active alginate films with amaranth phenolic compounds can be tailored to be used as food packaging material with potential antioxidant activity.

Effect of Chitosan Nanoparticles Incorporating Antioxidants from Salvia hispanica L. on the Amaranth Flour Films.

Research background: Amaranth (Amaranthus hypochondriacus) flour produces films with excellent barrier properties against water vapor, allowing food preservation, but the mechanical properties are poor compared to synthetic films. One strategy to improve these properties is the incorporation of nanoparticles. The particles can also serve as a vehicle for the addition of antioxidant agents into the films. The objective of this work is to optimize the formulation for the preparation of amaranth flour films treated with antioxidant chia (Salvia hispanica L.) extract-loaded chitosan particles using response surface methodology (RSM). Experimental approach: Chitosan nanoparticles with the extract were synthesized by ionic gelation, and the films were made by the casting method. Three independent variables were assigned: amaranth flour (4-6%), glycerol (25-35%) and chitosan nanoparticles loaded with the chia extract (0-0.75%). We then evaluated the physical (thickness), mechanical (tensile strength, Young´s modulus and elongation), barrier (water vapor permeability, moisture and water solubility) and antioxidant properties of the films. The experimental results of the properties were analyzed using a Box-Behnken experimental design generating 15 runs with three replicates at the central point. Results and conclusions: Second and third order polynomial models were obtained from the ANOVA analysis of the evaluated responses, and high coefficients of determination were found (0.91-1.0). The water vapor permeability of the films was 0.82-2.39·10-7 (g·mm)/(Pa·s·m2), tensile strength was 0.33-1.63 MPa and antioxidant activity 2.24-5.65%. The variables had different effects on the films: glycerol negatively affected their properties, and the permeability values increased with increased amaranth flour content. The nanoparticles improved the mechanical, barrier and antioxidant properties of the films compared to the films without nanosystems. The optimal formulation was 4% amaranth flour, 25% glycerol and 0.36% chitosan nanoparticles. The optimized films had better mechanical (1.62 MPa) properties, a low water vapor permeability value (0.91·10-7 (g·mm)/(Pa·s·m2)) and moderate antioxidant activity (6.43%). Novelty and scientific contribution: The results show the effect of chitosan nanoparticles on the properties of amaranth flour films for the first time. The resulting equations are useful in the design of food packaging.

Polyethylene Films Containing Plant Extracts in the Polymer Matrix as Antibacterial and Antiviral Materials.

Low density polyethylene (LDPE) films covered with active coatings containing mixtures of rosemary, raspberry, and pomegranate CO2 extracts were found to be active against selected bacterial strains that may extend the shelf life of food products. The coatings also offer antiviral activity, due to their influence on the activity of Φ6 bacteriophage, selected as a surrogate for SARS-CoV-2 particles. The mixture of these extracts could be incorporated into a polymer matrix to obtain a foil with antibacterial and antiviral properties. The initial goal of this work was to obtain active LDPE films containing a mixture of CO2 extracts of the aforementioned plants, incorporated into an LDPE matrix via an extrusion process. The second aim of this study was to demonstrate the antibacterial properties of the active films against Gram-positive and Gram-negative bacteria, and to determine the antiviral effect of the modified material on Φ6 bacteriophage. In addition, an analysis was made on the influence of the active mixture on the polymer physicochemical features, e.g., mechanical and thermal properties, as well as its color and transparency. The results of this research indicated that the LDPE film containing a mixture of raspberry, rosemary, and pomegranate CO2 extracts incorporated into an LDPE matrix inhibited the growth of Staphylococcus aureus. This film was also found to be active against Bacillus subtilis. This modified film did not inhibit the growth of Escherichia coli and Pseudomonas syringae cells; however, their number decreased significantly. The LDPE active film was also found to be active against Φ6 particles, meaning that the film had antiviral properties. The incorporation of the mixture of CO2 extracts into the polymer matrix affected its mechanical properties. It was observed that parameters describing mechanical properties decreased, although did not affect the transition of LDPE significantly. Additionally, the modified film exhibited barrier properties towards UV radiation. Modified PE/CO2 extracts films could be applied as a functional food packaging material with antibacterial and antiviral properties.

Active biopolymer films based on furcellaran, whey protein isolate and Borago officinalis extract: characterization and application in smoked pork ham production.

BACKGROUND: The meat industry is determined to find biodegradable packaging with properties similar to plastic. Furcellaran (FUR) and whey protein isolate (WPI) were used as a film matrix in which Borago officinalis extract (BOE) was incorporated as an antioxidant compound. The film's mechanical properties, water behavior, surface color, and antioxidant power were analyzed.Smoked hams were manufactured using two different types of film application: cured meat covered with film, smoked and cooked or hamsafter smoking, cooking and cooling. Smoked, vacuum packed ham was used as a control sample. The products were stored at 4 °C for 21 days and analyzed every 7 days. RESULTS: The elongation at break (EAB) and tensile strength (TS) of FUR/WPI films without the extract were 6.30% and 20.59 MPa, respectively, and after incorporating BOE, the EAB and TS were 24.30% and 15.33 MPa, respectively. The films with BOE were darker and had greater antioxidant capacity. The water content and activity in the products with films decreased along with storage time while the control remained stable. The results of microbiological, oxidation product accumulation, and sensory analysis were comparable in all the products. CONCLUSIONS: The smoking time can be reduced due to the dark color of the hams covered with BOE film. The barrier properties of those films should be increased. Other parameters were comparable to plastic packaging. The films therefore have the potential to be used instead of plastic packaging in the meat industry. © 2020 Society of Chemical Industry.

Effect of Sonication on the Properties of Flaxseed Gum Films Incorporated with Carvacrol.

Carvacrol is a natural compound known to be a highly effective antibacterial; however, it is a hydrophobic molecule, which is a limitation to its use within food packaging. Flaxseed gum (FG) films containing different contents of carvacrol (C) were produced by a film-casting method with sonication. The effects of sonication power and time on the properties of the FG-C films were investigated by measuring the film thickness, mechanical properties, contact angle, opacity, water vapor permeability (WVP), water sorption isotherm, Fourier transform infrared spectroscopy(FTIR), differential scanning calorimetry (DSC), antibacterial and antioxidant activities, and microstructure. The results showed that sonication power and time had significant effects on mechanical and barrier properties, film opacity, and degradability (p < 0.05). The tensile strength (TS) and elongation at break (EB) values exhibited an obvious improvement after sonication, and FG-0.5C-6030 had the lowest TS (33.40 MPa) and EB (4.46%) values. FG-C films formed a denser structure and the contact angle was improved as a result of sonication, which improved the integration of carvacrol into the FG matrix. In terms of microstructure, sonication resulted in a homogeneous and continuous crosssection of FG-C films, and regular surface and cross-sectional images were obtained through the highest acoustic intensity and longest time treatment. The FG films incorporated with carvacrol displayed antibacterial properties against Staphylococcus aureus, Vibrio parahaemolyticus, Shewanella putrefaciens, and Pseudomonas fluorescens, as well as increased antioxidant properties, and sonication was proven to enhance both of them.

Phytolacca americana extract as a quality-enhancing factor for biodegradable double-layered films based on furcellaran and gelatin - Property assessment.

For the first time, novel active double-layered films based on furcellaran (FUR) and gelatin (GEL) with the addition of Phytolacca americana (PA) extract were obtained. The 1st layer consisted of FUR and GEL, while the aqueous extract of P. americana berries was added in three different concentrations to the 2nd FUR-based layer. The films were characterised by good mechanical (TS range of 0.0011-0.0013 MPa, EAB range between 30.38 %-33.51 %) and water properties (WVTR range of 574.74-588.49 g/m2xd). Structural analysis (SEM and AFM) confirmed good film structure: regular, without cracks or air bubbles. The films showed antioxidant activity tested via the Folin-Ciocâlteu method (4.77-20.70 mg GAExg-1), FRAP assay (0.18-3.40 mM TExg-1) and CUPRAC assay (48.63-53.99 mM TExg-1). The film with the highest PA concentration (6 %) demonstrated the ability to neutralise free radicals, DPPH• and ABTS2+•, at the levels of 1.97 % and 17.34 %, respectively. The ecotoxicity test performed on Lepidium sativum seeds confirmed the lack of ecotoxic film aspects. The biodegradation test indicated that the films are biodegradable. The obtained films can be a good alternative to plastic packaging films (used in the food packaging industry), which are currently a global problem related to the development of post-consumer plastics.

Multifunctional packaging film with sustained release behavior triggered by pH microenvironment for efficient preservation of pork.

To tackle microbial contamination and spoilage during pork storage, transportation, and sales, a novel packaging film with exogenously driven controlled release behavior was designed through the demand-directed preparation of the emulsion of shellac-encapsulated cinnamaldehyde nano-capsules (SNCs) and the ingenious integration of chitosan (CS) film matrix. Among them, the SNCs were synthesized using the solvent exchange method, allowing controlled release of cinnamaldehyde (CA) in response to volatile alkaline substances present in the meat. The electrostatic interaction between SNCs and CS molecules improved the thermal stability, water resistance, tensile strength, and viscosity of the film-forming solution, while the CA in SNCs enhanced UV resistance and antimicrobial properties of the film. Notably, the CS film with SNC-100 loaded (CSNCs-100 film) achieved 99% inhibition against both E. coli and S. aureus, regulated environmental moisture, and prolonged the freshness duration of pork by an impressive six days.

Apple pectin-based active films to preserve oil: Effects of naturally branched phytoglycogen-curcumin host.

Pectin has excellent film-forming properties, but its functional properties need to be enhanced. Therefore, we constructed naturally branched phytoglycogen (PG) nanoparticles to solubilize curcumin (CCM) and further enhance the properties of apple pectin-based active films. The size of the PG spherical particles ranged from 30 to 100 nm with some aggregates. The branch density of the PG was 6.02 %. These PG nanoparticles increased the solubility of CCM nearly 1742-fold and a nanosized phytoglycogen-curcumin (PG-CCM) host was formed via hydrogen bonding and hydrophobic interaction. This host promoted the formation of pectin-based films with a dense structure and increased their tensile strength to 23.51 MPa. The coefficient to water vapor permeability, oxygen permeability and carbon dioxide permeability were all decreased indicating their barrier performance were improved. Among them, the oxygen permeability coefficient decreased most, from 1.14 × 10-7 g·m-1·s-1 to 0.8 × 10-7 g·m-1·s-1. Also, the transmittance of the active film at 280 nm and 660 nm decreased to 0.65 % and 72.10 %. Antioxidant and antibacterial properties were significantly enhanced (P < 0.05). And the results showed this film was an excellent oil packaging material. The active film incorporating PG-CCM host can replace heat-sealed plastic bags/pouch made from polyethylene and polypropylene synthetic plastics, and solve the problem that plastic packaging is difficult to degrade and cannot be squeezed clean. This provides a new conceptual framework for developing pectin-based active films by incorporating of PG and CCM.

Evaluation biodegradable films with green tea extract for interleafing sliced meat products.

A selection of formulations with different polymers and concentrations of green tea extract was conducted for application as interleafs in sliced meat products. Films were formulated using cellulose acetate, corn starch, and chitosan with the addition of 1.0, 2.5, and 5.0% green tea extract. Higher antioxidant activity was observed with the 1.0% concentration of green tea extract (P < 0.05), regardless of the formulation, with continuous release of the extract for up to 60 days and average IC50 of 0.09 and 0.31 mg/mL for the corn starch and chitosan active films, respectively. Interleafing the sliced ham resulted in lower lipid oxidation after 60 days of storage (P < 0.05). Starch-based films with green tea extract were effective, significantly reducing lipid oxidation in sliced and interleafed cooked ham, suggesting their potential to extend the shelf life of these refrigerated products.

Preparation and characterization of active films based on oregano essential oil microcapsules/soybean protein isolate/sodium carboxymethyl cellulose.

This study aimed to prepare oregano essential oil microcapsules (EOMs) by the active coalescence method using gelatin and sodium alginate as wall materials and oregano essential oil (OEO) as the core material. EOMs were added to the soybean protein isolate (SPI)/sodium carboxymethyl cellulose (CMC) matrix to prepare SPI-CMC-EOM active films, and the physical and chemical features of the active films and EOMs were characterized. The results showed that the microencapsulated OEO could protect its active ingredients. Scanning electron microscopy results showed that EOMs were highly compatible with the film matrix. The solubility of active films decreased upon adding EOMs, and their ultraviolet resistance and thermal stability also improved. When the added amount of EOMs was 5 %, the active films had the best mechanical properties and the lowest water vapor permeability. The active films prepared under this condition had excellent comprehensive performance. Also, adding EOMs considerably enhanced the antioxidant of the active films and endowed them with antibacterial properties. The application of the SPI-CMC-EOM films to A. bisporus effectively delayed senescence and maintained the freshness of the postharvest A. bisporus. This study provided a theoretical foundation for the incorporation of EOMs into active films based on biological materials.

Characterization of active films based on chitosan/polyvinyl alcohol integrated with ginger essential oil-loaded bacterial cellulose and application in sea bass (Lateolabrax japonicas) packaging.

The poor mechanical properties, low water-resistance, and limited antimicrobial activity of chitosan (CS)/polyvinyl alcohol (PVA) based film limited its application in aquatic product preservation. Herein, bacterial cellulose (BC) was used to load ginger essential oil (GEO). The effects of the addition of BC and different concentrations of GEO on the physicochemical and antimicrobial activities of films were systematically evaluated. Finally, the application of sea bass fillets was investigated. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction analysis (XRD) analysis indicated dense networks were formed, which was verified by enhanced physical properties. The mechanical properties, barrier properties, and antimicrobial activities enhanced as GEO concentration increased. CPB0.8 (0.8 % GEO) film had better tensile strength (TS) and barrier performance, improved the quality, and extended the shelf-life of sea bass for another 6 days at least. Overall, active films are potential packaging materials for aquatic products.

Choline chloride-based deep eutectic solvents as plasticizer and active agent in chitosan films.

The growing concern over extending the shelf life of food products, coupled with the escalating environmental impact of synthetic plastic waste, has fuelled a quest for bio-based alternatives in packaging research. In response to this pressing need, our study delves into the synthesis of chitosan-based films incorporating a deep eutectic solvents (DES). Choline chloride and diverse hydrogen bond donors were used as plasticizers, we also explored the active properties of DES integrated into the chitosan (Ch) matrix. The Ch-based films with chlorine chloride: citric acid can prevent the mold spotting up to 29 days longer in comparison to bread wrapped in polyethylene films (PE). The obtained Ch/DES films exhibited mechanical properties comparable to conventional PE (e.g., up to tensile strength of 26 MPa and up to 210% in case of elongation at break). This synthesis approach represents a significant stride towards environmentally friendly packaging materials, aligning with the principles of green chemistry.

Physicochemical properties of active films of rose essential oil produced using soy protein isolate-polyphenol conjugates for cherry tomato preservation.

Soy protein isolate (SPI)-polyphenol conjugates were produced by grafting SPI individually with curcumin, naringenin, and catechin. The resulting conjugates showed better emulsifying properties and were used to develop active films containing rose essential oil. The effect of conjugation on the physicochemical and mechanical properties of these emulsion-based films was evaluated. The results showed that the barrier and mechanical properties of the films were improved when the SPI-polyphenol conjugates were used to emulsify the essential oil; in particular, the SPI-curcumin conjugate showed significant improvement. The improvements on the water vapor and oxygen barrier properties in the films were attributed to the formation of compact structure. Emulsion-based films stabilized by SPI-polyphenol conjugates showed antioxidant and antibacterial activities. They also demonstrated an ability to extend the shelf life of cherry tomatoes, as indicated by better preservation of weight, firmness, and ascorbic acid content.

Improving the Properties of Gelatin-Based Films by Incorporation of "Pitanga" Leaf Extract and Crystalline Nanocellulose.

Biopolymer-based films can be activated by the incorporation of active compounds into their matrix. Plant extracts are rich in phenolic compounds, which have antimicrobial and/or antioxidant properties. The aim of this study was to produce gelatin-based active films and nanocomposite films incorporated with "pitanga" (Eugenia uniflora L.) leaf extract (PLE) and/or crystalline nanocellulose extracted from soybean straw (CN), and to study the physicochemical, functional, microstructural, thermal, UV/Vis light barrier, and antioxidant properties of these materials. PLE enhanced some film properties, such as tensile strength (from 30.2 MPa to 40.6 MPa), elastic modulus (from 9.3 MPa to 11.3 MPa), the UV/Vis light barrier, and antioxidant activity, in addition to affecting the microstructural, surface, and color properties. These improvements were even more significant in nanocomposites simultaneously containing PLE and CN (59.5 MPa for tensile strength and 15.1 MPa for elastic modulus), and these composites also had lower moisture content (12.2% compared to 13.5-14.4% for other treatments) and solubility in water (from 48.9% to 44.1%). These improvements may be the result of interactions that occur between PLE's polyphenols and gelatin, mainly in the presence of CN, probably due to the formation of a stable PLE-CN-gelatin complex. These results are relevant for the food packaging sector, as the activated nanocomposite films exhibited enhanced active, barrier, and mechanical properties due to the presence of PLE and CN, in addition to being entirely produced with sustainable components from natural and renewable sources.

Konjac glucomannan-based highly antibacterial active films loaded with thyme essential oil through bacterial cellulose nanofibers/Ag nanoparticles stabilized Pickering emulsions.

The aim of this study was to develop novel konjac glucomannan (KGM)-based highly antibacterial active films, where five types of films were prepared and compared. The microstructure results showed that KGM-based films loaded with thyme essential oil (TEO) through bacterial cellulose nanofibers/Ag nanoparticles (BCNs/Ag nanoparticles) stabilized Pickering emulsions (Type V films) displayed the smoothest surface and the most evenly dispersed TEO droplets as compared with the other four types of films. Moreover, Type V films showed the highest contact angle value (86.28°), the best thermal stability and mechanical properties. Furthermore, Type V films presented the highest total phenol content (13.23 mg gallic acid equivalent/g film) and the best antioxidant activity (33.96 %) as well as the best sustained-release property, thus showing the best antibacterial activity, which was probably due to that BCNs/Ag nanoparticles and TEO displayed a synergistic effect to some extent. Consequently, Type V film-forming solutions were used as coatings for tangerines. The results showed that the tangerines treated with Type V coatings displayed excellent fresh-keeping properties. Therefore, the coatings, KGM-based film-forming solutions loaded with TEO through BCNs/Ag nanoparticles stabilized Pickering emulsions, have great potential for the preservation of fruits and vegetables.

Films Based on Biopolymers Incorporated with Active Compounds Encapsulated in Emulsions: Properties and Potential Applications-A Review.

The rising consumer demand for safer, healthier, and fresher-like food has led to the emergence of new concepts in food packaging. In addition, the growing concern about environmental issues has increased the search for materials derived from non-petroleum sources and biodegradable options. Thus, active films based on biopolymers loaded with natural active compounds have great potential to be used as food packaging. However, several lipophilic active compounds are difficult to incorporate into aqueous film-forming solutions based on polysaccharides or proteins, and the hydrophilic active compounds require protection against oxidation. One way to incorporate these active compounds into film matrices is to encapsulate them in emulsions, such as microemulsions, nanoemulsions, Pickering emulsions, or double emulsions. However, emulsion characteristics can influence the properties of active films, such as mechanical, barrier, and optical properties. This review addresses the advantages of using emulsions to encapsulate active compounds before their incorporation into biopolymeric matrices, the main characteristics of these emulsions (emulsion type, droplet size, and emulsifier nature), and their influence on active film properties. Furthermore, we review the recent applications of the emulsion-charged active films in food systems.

Preparation and characterization of carboxymethyl chitosan/pullulan composite film incorporated with eugenol and its application in the preservation of chilled meat.

To solve the problem of easy spoilage of chilled meat during storage, we fabricated a novel composite film using carboxymethyl chitosan (CMCS)/pullulan (Pul)/eugenol (E) by casting method. The results showed that the mechanical properties of the films were better when the CMCS/Pul ratio was 2.5/2.5. The Fourier transform infrared spectroscopy (FTIR) results showed that intermolecular hydrogen bonds were formed among E, CMCS, and Pul, which was consistent with the rheological test results. Scanning electron microscopic (SEM) images showed that eugenol was well dispersed in the CMCS/Pul matrix. The addition of eugenol significantly increased the antibacterial properties and antioxidant properties. Moreover, when 5% eugenol was added, the water vapor permeability (WVP) of the film reduced to 2.41 × 10-11 g/m·s·Pa. Finally, the freshness of the chilled meat wrapped with the eugenol-containing composite film was prolonged, thereby offering a potential alternative to synthetic materials.

Physicochemical and antioxidant properties of tannic acid crosslinked cationic starch/chitosan based active films for ladyfinger packaging application.

In the present study, cationic starch (CS)/chitosan (CH) incorporated with tannic acid (TA)(CSCT) eco-friendly films were prepared by employing an inexpensive solvent casting technique. Influence of TA on the physicochemical and antimicrobial properties of CS/CH polymer matrix were studied. The FTIR findings and homogeneous, dense SEM micrographs confirms the effective interaction of TA with CS/CH polymer matrix. CSCT-3 active film displayed tensile strength of 26.99±1.91 MPa, which is more substantial than commercially available polyethylene (PE) (12-16 MPa) films. The active films exhibited excellent barrier properties against moisture and water, supported by increased water contact angle values (86.97±0.29°). Overall migration rate of active films was found to be below the permitted limit of 10mg/dm2. The active films showed around 56% of degradation in soil within 15 days. Besides, the active films showed concurring impact against food borne pathogens like E. coli, S. aureus and C. albicans. The CSCT-3 active film presented 90.83% of antioxidant capacity, demonstrating the effective prevention of food oxidation related deterioration. Ladyfinger packaging was inspected to examine the ability of active films as packaging material resulted in effectively resisting deterioration and extending shelf life in comparison with traditional PE packaging.

Preservative effect of gelatin/chitosan-based films incorporated with lemon essential oil on grass carp (Ctenopharyngodon idellus) fillets during storage.

The present study investigated the effect of fish gelatin/chitosan-based (FG/CS-based) films incorporated with lemon essential oil (LEO) on grass carp fillets in terms of moisture status, total volatile basic nitrogen (TVB-N), and microbial community succession during chilled (4 °C) and iced (0 °C) storage. Low-field nuclear magnetic resonance (LF-NMR) revealed that the active films remarkably inhibited moisture transformation from being the immobilized to free water in grass carp fillets, accompanied with the reduced T22 relaxation time. Besides, magnetic resonance imaging (MRI) detected a higher density of proton in the treated fish samples, indicating that the active films could improve the water-holding capacity of fish samples. Moreover, high-throughput 16S rRNA sequencing suggested that the FG/CS-based films loaded with LEO efficiently decreased the relative abundance of the bacterial genera Shewanella and Aeromonas in grass carp fillets, with minimal accumulation of TVB-N during storage. Additionally, the low storage temperature (0 °C) could further enhance the preservative effect of the active films on the fish samples, which together prolonged their shelf-life to 18 days. Overall, the combination of the active films and iced storage could provide a promising strategy to preserve grass carp fillets.