Antibacterial Activity and Physical Properties of Edible Chitosan Films Exposed to Low-pressure Plasma

The challenge for food industry is developing gentle processes concept, which will prevent food spoilage and leave a food natural, minimally processed, fresh-like and safe. A new technique of food preservation could be usage of combined methods of green process, such as cold gas plasma with bioactive substances, and protective coatings. The aim of this study was to determine the antibacterial activity of chitosan films incorporated with lysozyme exposed to helium plasma treatment as well as evaluate their physical properties. The edible films have been prepared basing on low molecular weight chitosan by casting from lactic acid solution with water solution of lysozyme in three various concentrations (0, 0.5 and 1 %). Dried films were then modified by exposition on cold helium plasma treatment for 0, 5 and 10 min. Obtained films were tested against growth of Listeria monocytogenes, Yersinia enterocolitica and Pseudomonas fluorescens. In order to characterize chitosan-based films, their mechanical properties, theromogravimetric analysis (TGA), dynamic mechanical thermal analysis (DMTA), contact angle measurement, water vapour permeability (WVP) and scanning electron microscopy (SEM) were evaluated. The films with 1 % lysozyme incorporation enhanced the inhibition efficiency of chitosan-based films against gram-positive (L. monocytogenes) and gram-negative (P. fluorescens) bacteria, where reduction zones were 42.5 and 69.8 mm, respectively. Besides that, hydrolytic changes of chitosan chain caused by lysozyme activity were confirmed by TGA and DMTA. Contact angles and WVP of tested films were not significantly affected by helium plasma exposition, nor lysozyme dosage. Hydrophilic natures of chitosan-based films were confirmed by both tests. Microscopy image of cross-section structure was smooth and continuous due to lysozyme addition in film composition. Application of chitosan films incorporated with lysozyme and low-pressure plasma treatment could be used as innovative preservation method in a wide range of food products.     

Optimization of the biocide properties of chitosan for its application in the design of active films of interest in the food area

The influence on biocide performance of some unprecedented physicochemical features of chitosan cast films such as film thickness, pH of the nutrient broth, film neutralization, film autoclave sterilization and temperature exposure were analyzed against Staphylococcus aureus and in some experiments also against Salmonella spp. The work demonstrates for the first time the influence of the release or positive migration of protonated glucosamine fractions from the biopolymer into the microbial culture as the responsible event for the antimicrobial performance of the biopolymer under the studied conditions. From the results, a reliable and reproducible method for the determination of the bactericidal activity of chitosan-based films was developed in an attempt to standardize the testing conditions for the optimum design of active antimicrobial food packaging films and coating applications.     

Development and evaluation of a novel biodegradable film made from chitosan and cinnamon essential oil with low affinity toward water

Combining antimicrobial agents such as plant essential oils directly into a food packaging is a form of active packaging. In this work chitosan-based films containing cinnamon essential oil (CEO) at level of 0.4%, .0.8%, and 1.5% and 2% (v/v) were prepared to examine their antibacterial, physical and mechanical properties. Scanning electron microscopy was carried out to explain structure–property relationships. Incorporating CEO into chitosan-based films increased antimicrobial activity. CEO decreased moisture content, solubility in water, water vapour permeability and elongation at break of chitosan films. It is postulated that the unique properties of the CEO added films could suggest the cross-linking effect of CEO components within the chitosan matrix. Electron microscopy images confirmed the results obtained in this study.     

壳聚糖复合膜在果蔬保鲜中的应用

壳聚糖具有良好的生物相容性、可生物降解性、安全性、成膜性、抗菌性等,常用于果蔬保鲜。但由于机械性能和生物活性不足,为了增强其理化性能和生物活性,常添加生物聚合物、抗菌剂、抗氧化剂等功能成分以提高膜综合性能。综述了壳聚糖膜和壳聚糖/多糖、壳聚糖/蛋白质、壳聚糖/脂质、壳聚糖/抗菌剂、壳聚糖/抗氧化剂等壳聚糖基复合膜在果蔬保鲜中的应用进展,从壳聚糖的诱导活性、成膜特性和抗菌活性三个方面总结了壳聚糖膜的保鲜机理,分析了壳聚糖基复合膜目前在果蔬保鲜实际应用中存在的问题,并对未来发展方向进行了展望,以期为开发安全、高效、绿色、经济的壳聚糖基果蔬保鲜膜提供一定的理论指导。     

Effects of gelatin origin,bovine-hide and tuna-skin,on the properties of compound gelatin–chitosan films

With the purpose to improve the physico-chemical performance of plain gelatin and chitosan films, compound gelatin–chitosan films were prepared. The effect of the gelatin origin (commercial bovine-hide gelatin and laboratory-made tuna-skin gelatin) on the physico-chemical properties of films was studied. The dynamic viscoelastic properties (elastic modulus G′, viscous modulus, G″ and phase angle) of the film-forming solutions upon cooling and subsequent heating revealed that the interactions between gelatin and chitosan were stronger in the blends made with tuna-skin gelatin than in the blends made with bovine-hide gelatin. As a result, the fish gelatin–chitosan films were more water resistant (∼18% water solubility for tuna vs 30% for bovine) and more deformable (∼68% breaking deformation for tuna vs 11% for bovine) than the bovine gelatin–chitosan films. The breaking strength of gelatin–chitosan films, whatever the gelatin origin, was higher than that of plain gelatin films. Bovine gelatin–chitosan films showed a significant lower water vapour permeability (WVP) than the corresponding plain films, whereas tuna gelatin–chitosan ones were only significantly less permeable than plain chitosan film. Complex gelatin–chitosan films behaved at room temperature as rubbery semicrystalline materials. In spite of gelatin–chitosan interactions, all the chitosan-containing films exhibited antimicrobial activity against Staphylococcus aureus, a relevant food poisoning. Mixing gelatin and chitosan may be a means to improve the physico-chemical performance of gelatin and chitosan plain films, especially when using fish gelatin, without altering the antimicrobial properties.     

Effects of drying methods and conditions on antimicrobial activity of edible chitosan films enriched with galangal extract

The aim of this work was to study the effects of drying methods and conditions (i.e., ambient drying, hot air drying at 40 °C, vacuum drying and low-pressure superheated steam drying within the temperature range of 70–90 °C at an absolute pressure of 10 kPa) as well as the concentration of galangal extract on the antimicrobial activity of edible chitosan films against Staphylococcus aureus. Galangal extract was added to the film forming solution as a natural antimicrobial agent in the concentration range of 0.3–0.9 g/100 g. Fourier transform infrared (FTIR) spectra and swelling of the films were also evaluated to investigate interaction between chitosan and the galangal extract. The antimicrobial activity of the films was evaluated by the disc diffusion and viable cell count method, while the morphology of bacteria treated with the antimicrobial films was observed via transmission electron microscopy (TEM). The antimicrobial activity, swelling and functional group interaction of the antimicrobial films were found to be affected by the drying methods and conditions as well as the concentration of the galangal extract. The electron microscopic observations revealed that cell wall and cell membrane of S. aureus treated by the antimicrobial films were significantly damaged.     

Preparation and Characterization of Antimicrobial Films Based on Chitosan for Active Food Packaging Applications

The aim of this paper was to characterize chitosan samples from the shrimp shells for the later development of antimicrobial active systems. These systems include 100 % chitosan-based films obtained by casting, polyamide films with 5 and 10 % of chitosan obtained by extrusion and polyethylene/polyethylene terephthalate films with a coating of 0.6 % of chitosan. For that purpose, several analytical techniques including IR, ¹H NMR, GPC, and microscopic techniques (scanning electron microscopy and transmission electron microscopy) were used. Within the studied samples, C1 showed the lowest DA and MW and consequently presented the most suitable properties for the development of an active packaging. Additionally, mechanical properties were performed. The effectiveness of the developed systems was evaluated by means of microbiological assays. The tested films showed antimicrobial capacity against coliform enterobacteria, mesophilic aerobic microorganism, and yeast and moulds.     

The antimicrobial,mechanical, physical and structural properties of chitosan–gallic acid films

Chitosan films incorporated with various concentrations of gallic acid were prepared and investigated for antimicrobial, mechanical, physical and structural properties. Four bacterial strains that commonly contaminate food products were chosen as target bacteria to evaluate the antimicrobial activity of the prepared gallic acid–chitosan films. The incorporation of gallic acid significantly increased the antimicrobial activities of the films against Escherichia coli, Salmonella typhimurium, Listeria innocua and Bacillus subtilis. Chitosan films incorporated with 1.5 g/100 g gallic acid showed the strongest antimicrobial activity. It was also found that tensile strength (TS) of chitosan film was significantly increased when incorporating 0.5 g/100 g gallic acid. Inclusion of 0.5 g/100 g gallic acid also significantly decreased water vapor permeability (WVP) and oxygen permeability (OP). Microstructure of the films was investigated by Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) and it was found that gallic acid was dispersed homogenously into the chitosan matrix.     

Characterization of salmon gelatin based film on antimicrobial properties of chitosan against E. coli

Salmon gelatin and chitosan are high potential biopolymer to obtain edible films with antimicrobial effect for fresh meat. Therefore, it is important to characterize the structural (glassy or rubbery) state of the gelatin film on antimicrobial properties of chitosan. Extracted salmon (Salmo Salar) gelatin (acid-basic extraction) and low molecular weight chitosan solutions (0; 0.25; 0.5 and 1% w/w) at pH 5.5 were prepared to obtain a final concentration to 7% w/w (gelatin + chitosan). Films were obtained by casting at 5 °C and equilibrated at 33% and 85% of relative humidity (RH) in order to obtain a glassy and rubbery state, respectively. Water content, specific volume and glass transition temperature (Tg) were measured and antimicrobial properties against Escherichia coli (E.c.) (105 ufc/ml) were obtained by agar plate diffusion and kinetic measurements at 37 °C. Chitosan diffusion in agar plate was determined using liquid solutions and films over agar plate. The results showed diminution of Tg (maintaining structural state) as increase chitosan concentration, increasing also specific volume and water content due their high capacity to adsorb water. Although inhibition by chitosan of bacteria growth kinetic was 100%, solutions and films samples could not diffuse in the agar. In conclusion, the chitosan could not diffuse on the films matrix and also in agar plate and therefore in food when is combined with salmon gelatin. The implication of this work have been investigate the use of edible films from marine sources with antimicrobial effects in an effort to growing demand from consumers for safer and better quality foods.     

Physical and antimicrobial properties of chitosan–tea tree essential oil composite films

Antimicrobial films were prepared by incorporating different concentrations of tea tree essential oil (TTO) into chitosan (CH) films. Film-forming dispersions (FFD) were characterized in terms of rheological properties, particle size distribution and ζ-potential. In order to study the impact of the incorporation of TTO into the CH matrix, the water vapour permeability (WVP), mechanical and optical properties of the dry films were evaluated. The properties of the films were related with their microstructure, which was observed by SEM. Furthermore, the antimicrobial effectiveness of CH–TTO composite films against Listeria monocytogenes and Penicillium italicum was studied.     

Biodegradable gelatin–chitosan films incorporated with essential oils as antimicrobial agents for fish preservation

Essential oils of clove (Syzygium aromaticum L.), fennel (Foeniculum vulgare Miller), cypress (Cupressus sempervirens L.), lavender (Lavandula angustifolia), thyme (Thymus vulgaris L.), herb-of-the-cross (Verbena officinalis L.), pine (Pinus sylvestris) and rosemary (Rosmarinus officinalis) were tested for their antimicrobial activity on 18 genera of bacteria, which included some important food pathogen and spoilage bacteria. Clove essential oil showed the highest inhibitory effect, followed by rosemary and lavender. In an attempt to evaluate the usefulness of these essential oils as food preservatives, they were also tested on an extract made of fish, where clove and thyme essential oils were the most effective. Then, gelatin–chitosan-based edible films incorporated with clove essential oil were elaborated and their antimicrobial activity tested against six selected microorganisms: Pseudomonas fluorescens, Shewanella putrefaciens, Photobacterium phosphoreum, Listeria innocua, Escherichia coli and Lactobacillus acidophilus. The clove-containing films inhibited all these microorganisms irrespectively of the film matrix or type of microorganism. In a further experiment, when the complex gelatin–chitosan film incorporating clove essential oil was applied to fish during chilled storage, the growth of microorganisms was drastically reduced in gram-negative bacteria, especially enterobacteria, while lactic acid bacteria remained practically constant for much of the storage period. The effect on the microorganisms during this period was in accordance with biochemical indexes of quality, indicating the viability of these films for fish preservation.     

Antimicrobial activity and physical properties of chitosan–tapioca starch based edible films and coatings

Antimicrobial activity of edible coating solutions based on chitosan and blends of chitosan–tapioca starch with or without potassium sorbate (KS) addition was studied. The agar well diffusion assay showed an antagonist effect on the efficiency of chitosan against Lactobacillus spp. when KS and/or tapioca starch were present. A salmon slice coating assay showed that the chitosan solution was the best coating since aerobic mesophilic and psychrophilic cell counts were reduced, pH and weight loss remained acceptable throughout refrigerated storage, extending global quality to 6-days. Chitosan–tapioca starch based films reduced Zygosaccharomyces bailii external spoilage in a semisolid product but were not effective against Lactobacillus spp. The results suggest that antibacterial action depended on the application technique, due to the fact that chitosan is more available in a coating solution than in a film matrix. Interactions between chitosan–starch and/or KS could affect film physical properties and the antimicrobial activity of chitosan. The addition of chitosan reduced water vapor permeability and solubility of starch films.     

Chitosan application for active bio-based films production and potential in the food industry: Review

During the past decade, there was an increasing interest to develop and use bio-based active films which are characterized by antimicrobial and antifungal activities in order to improve food preservation and to reduce the use of chemical preservatives. Biologically active bio-molecules such as chitosan and its derivatives have a significant potential in the food industry in view of contaminations associated with food products and the increasing concerns in relation with the negative environmental impact of conventional packaging materials such as plastics. Chitosan offers real potential for applications in the food industry due to its particular physico-chemical properties, short time biodegradability, biocompatibility with human tissues, antimicrobial an antifungal activities, and non-toxicity. Thus, chitosan-based films have attracted serious attention in food preservation and packaging technology. This is mainly due to a fact that chitosan exhibits high antimicrobial activity against pathogenic and spoilage micro-organisms, including fungi, and both Gram-positive and Gram-negative bacteria. The aim of the present review was to summarize the most important information on chitosan from its bioactivity point of view and to highlight various preparative methods used for chitosan-based active bio-films and their potential for applications in the food preservation and packaging technology.     

Influence of α-tocopherol on physicochemical properties of chitosan-based films

Chitosan has been exploited as a material for the development of edible films, and additionally can be used as a carrier of functional compounds such as α-tocopherol. The aim of this work was to evaluate the effects of the incorporation of α-tocopherol in chitosan-based films. FTIR and thermal analyses were performed and showed that the incorporation of α-tocopherol affects the chemical structure of chitosan-based films with the establishment of new chemical bonds and the decrease of crystallinity. Results also showed that the increase of α-tocopherol concentration promotes a decrease of water content (from 12.6 to 11.4%) of the films. The addition of α-tocopherol to chitosan films leads to a significant reduction (p < 0.05) of tensile strength from 34.06 to 16.24 MPa, and elongation-at-break from 53.84 to 23.12%. Film opacity values (ranging from 4.74 to 7.83%) increased when α-tocopherol was incorporated into the film. Antioxidant capacity of chitosan-based films was evaluated and was enhanced when α-tocopherol was present in the film matrix. Results showed that α-tocopherol can be successfully added to the chitosan films enhancing the final quality and shelf-life extension of food products.     

Chitosan Polymer as Bioactive Coating and Film against Aspergillus niger Contamination

ABSTRACT: The inhibitory activity of chitosan-based edible coatings and films was assessed against the Aspergillus niger food pathogen and deterioration microorganism. Spore-counting assays showed an almost total inhibition of A. niger growth when either film-forming solution or film were used at a low concentration of chitosan (0.1% w/v). Epifluorescence microscopic results showed the action of chitosan on the relative proportion of RNA compared with DNA. The water vapor permeability (WVP) of chitosan film was relatively low compared with the poor moisture barrier of some polysaccharide films. Moreover, a coating with chitosan film on an agar gel, used as a food model, induced a 30% reduction in water loss. These results showed potential applications of chitosan-based films as bioactive packaging with properties to limit the food dehydration phenomenon.     

Effects on Salmonella shell contamination and trans-shell penetration of coating hens' eggs with chitosan

Chitosan is a biopolymer with antimicrobial activity and film-forming properties. In this study, the effects on Salmonella shell contamination and trans-shell penetration of coating hens' eggs with chitosan was evaluated. A chitosan was selected from eight types (four non-commercial and four commercial) based on its antimicrobial activity against Salmonella enterica serovar Enteritidis (S. Enteritidis). For this purpose, a contact plate method was developed and chitosans were applied at a concentration of 0.25% (w/v). A commercial type with a molecular weight of 310-375 kDa and a deacetylation degree of 75% that reduced S. Enteritidis by 0.71 log₁₀ colony forming units compared to the control (without chitosan) was selected for further studies. The chitosan was shown to have antimicrobial activity against other egg borne bacteria, i.e., Acinetobacter baumannii, Alcaligenes sp., Carnobacterium sp., Pseudomonas sp., Serratia marcescens and Staphylococcus warneri, and against S. enterica serovar Typhimurium, Escherichia coli and Listeria monocytogenes. The effects of various concentrations of the selected chitosan (0.25%, 1% and 2%) on Salmonella shell contamination and trans-shell penetration were assessed using the agar molding technique. Effective reduction of eggshell contamination could not be demonstrated, but trans-shell penetration was significantly reduced in the presence of a 2% chitosan eggshell coating, with only 6.1% of the eggs being penetrated compared to 24.5% of the uncoated eggs. It was concluded that the 2% chitosan coating has the potential to reduce contamination of egg contents resulting from trans-shell penetration by S. Enteritidis.     

Antimicrobial compression-moulded wheat gluten films containing potassium sorbate

Antimicrobial glycerol-plasticized wheat gluten (WG) films containing potassium sorbate (PS) were successfully produced by compression moulding; a thermoplastic process involving high temperature and high pressure. Antifungal properties of the films were tested against Aspergillus niger and Fusarium incarnatum by the agar diffusion assay. The results indicated that films containing more than 10 wt.% PS showed antimicrobial activity against A. niger while films containing 2.5 wt.% or more of PS showed antimicrobial activity against F. incarnatum. It was also found that when the film was exposed to an absorbing medium (the agar solution), most of the PS was released, an interesting feature for edible active packaging. Despite the loss, a very promising result was that, without seeding of spores, the films resisted microbial growth for at least one week when the films were left in the agar solution. X-ray diffraction and field emission scanning electron microscopy revealed that the PS crystals were dissolved in the wheat gluten material. In addition to the antimicrobial properties, dynamic mechanical, tensile, PS loss, water vapour transmission rate and oxygen permeability data also indicated that PS acted as a plasticiser in the wheat gluten film.     

Silver/chitosan-based Janus particles: Synthesis,characterization, and assessment of antimicrobial activity in vivo and vitro

Janus particles containing chitosan and silver were synthesized in an eco-friendly manner and were confirmed using transmission electron microscopy. Based on the data of the antimicrobial activity assessment, this material exhibited a higher antimicrobial activity than virgin chitosan with long–lasting antibacterial effectiveness against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Salmonella choleraesuis bacteria, as well as Botrytis cinerea fungi. The results showed that the Janus polymer could completely suppress the growth and germination of B. cinerea at a concentration of 0.02 mg/mL in vitro and in vivo. This Janus polymer is an advanced functional material that combines the suitable properties of both components and could be an alternative new antimicrobial agent due to its unique chemical properties and pronounced antimicrobial activity. This material is a potential candidate for use in the food industry to prevent microbial contamination and to inhibit the growth of microorganisms, enhancing product quality and, extend shelf-life of fresh and processed agri-food products.     

Active chitosan–polyvinyl alcohol films with natural extracts

Active films were prepared from chitosan (Ch) and polyvinyl alcohol (PVA) containing aqueous mint extract (ME)/pomegranate peel extract (PE). The effect of these extracts on the physical, mechanical, antimicrobial and antioxidant properties of the films were studied. Increased protection against UV light was observed in the films containing the extracts. Addition of ME/PE improved the tensile strength of the films without affecting their puncture strength. Ch–PVA films incorporated with PE had the highest tensile strength (41.07 ± 0.88 MPa). Permeability characteristics of the films were not altered due to addition of extracts. ME/PE conferred antioxidant properties to Ch–PVA films as determined by DPPH radical scavenging activity. The films also exhibited antibacterial activity against Staphylococcus aureus and Bacillus cereus. PE containing films totally inhibited the growth of B. cereus and reduced the number of S. aureus by 2 log cycles. These results suggest that Ch–PVA film containing ME/PE can be used for development of active food packaging materials.     

Use of the jabuticaba (Myrciaria cauliflora) depulping residue to produce a natural pigment powder with functional properties

The aim of this study was to produce a natural pigment powder with functional properties. To optimise the spray-drying process, a central composite design with 17 treatments was used, in which the independent variables were the inlet drying air temperature (138–202 °C), the feed flow rate (20–67 mL/min), and the concentration of the carrier (maltodextrin, 100–300 g/kg of extract). The dependent variables were the moisture content, hygroscopicity, and anthocyanin retention. The samples were analysed by: anthocyanin and phenolic content; antioxidant properties based on the ORAC and DPPH assays; antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, Salmonella enteritidis, and Escherichia coli; and inhibitory activity against arginase, an enzyme produced by Leishmania amazonensis. The experimental design was not significant or predictive in the ranges studied. The selected samples contained high concentrations of anthocyanins and phenolics, and high antioxidant capacities. In addition, they exhibited antimicrobial activity against three of the four microorganisms tested, and some of the samples also exhibited a bactericidal effect. Moreover, the powders exerted inhibitory activity (64–69%) against arginase. The results obtained in this study suggest that the jabuticaba depulping residue could be used to produce a natural pigment with functional properties.