Development and characterization of antimicrobial poly(l-lactic acid) containing trans-2-hexenal trapped in cyclodextrins

Trans-2-hexenal, a naturally occurring plant volatile with antimicrobial capacity, was encapsulated into β-cyclodextrins (β-CDs), enzymatically modified starch, and shown effective to control main microorganisms causing food spoilage (Alternaria solani, Aspergillus niger, Botrytis cinerea, Colletotrichum acutatum, Penicillium sp). Loaded β-CDs were incorporated into a poly(l-lactic acid) (PLA) matrix by extrusion and casting, and yielded antimicrobial polymers made from natural resources. A masterbatch was used prior to sheet casting to improve the dispersion of the antimicrobial agent in the PLA matrix. However, this increased the number of extrusion processes for the material. The concentration of the antimicrobial compound in the polymers and its antimicrobial capacity against one food spoilage microorganism (A. solani) were measured during the different processing operations. Although the concentration of trans-2-hexenal was reduced by processing by about 70 and 99% compared to the loaded β-CDs, for the masterbatch and sheet, respectively, the polymers were still effective in reducing microbial growth. The changes of the polymer properties due to the addition of the antimicrobial agent were investigated, too. It was found that the mechanical and barrier properties of the PLA were changed (decreased by about half the tensile strength and elongation at break and nine-fold increased permeability) while the physical properties remained the same. Based on these results, the developed polymer may be a viable antimicrobial material for applications in food packaging.     

Application of β-Cyclodextrin/2-Nonanone Inclusion Complex as Active Agent to Design of Antimicrobial Packaging Films for Control of <Emphasis Type="Italic">Botrytis cinerea</Emphasis>

The aim of this study was to develop, characterize and evaluate in vitro the efficacy of active films, based on an inclusion complex formed by β-cyclodextrin, 2-nonanone and two polymer matrices (polylactic acid and low density polyethylene). The different films were characterized by scanning electronic microscopy (SEM), differential scanning calorimetry (DSC), themogravimetric analysis (TGA), optical properties and antimicrobial activity against B. cinerea. The results showed important differences in the parameters evaluated where the level of agglomerates of additives was a key to explain these changes. Finally, microbiological analysis showed high effectiveness in reducing the Botrytis cinerea growth. The active films developed in this study were able to inhibit the growth of phytopathogenic fungus B. cinerea at different experimental conditions. The studied films have potential use for packaging fresh fruit susceptible to biological attack by this fungus.     

Physical and antioxidant properties of chitosan and methylcellulose based films containing resveratrol

New trends in edible films focus on the improvement of their functionality through the incorporation of active compounds, such as antimicrobial or antioxidant agents. Resveratrol is a natural antioxidant found in a variety of plant species, such as grapes, and could be used for minimizing or preventing lipid oxidation in food products, retarding the formation of oxidation products, maintaining nutritional quality and prolonging the food shelf life. The aim of this work was to develop and characterize two different polymeric composite films (made with chitosan (CH) and methylcellulose (MC)) containing different amounts of resveratrol. This compound could be incorporated efficiently into both films, but provoke structural changes in the matrices, which became less stretchable (65–70% reduction of deformation at break at the greatest resveratrol content) and resistant to fracture (26 and 54% reduction of tensile at break for MC and CH, respectively, at the greatest resveratrol content) more opaque (significant reduction of the internal transmittance) and less glossy (about 60–65% reduction of gloss at the greatest resveratrol content). Film barrier properties were hardly improved by the presence of resveratrol; water vapour and oxygen permeability tend to slightly decrease when resveratrol was incorporated into both polymers. Composite films showed antioxidant activity, which was proportional to the resveratrol concentration in the film. None of the films showed antimicrobial activity against Penicillium italicum and Botrytis cinerea. Thus, these films could be applied to food products which are sensitive to oxidative processes to prolong their shelf life.     

Combined effect of high pressure treatment and anti-microbial bio-sourced materials on microorganisms' growth in model food during storage

Antimicrobial bio-sourced films based on poly(lactic acid) containing either carvacrol or allyl isothiocyanate (AITC) were prepared and their antimicrobial properties were assessed on Botrytis cinerea during storage and after a high pressure (HP) “pasteurisation-like” treatment (up to 800 MPa at ambient temperature). A dry process (extrusion + thermomoulding) was used to shape the material. The high temperature encountered during film processing dramatically decreased the carvacrol and AITC content in the film, leading to a less efficient antimicrobial activity. The use of β-cyclodextrin (β-CD) to encapsulate the active compounds before film processing proved to be efficient to protect the AITC against thermal degradation and to control its release from the films during its use. PLA-based films containing either AITC or β-CD encapsulated AITC showed a significant activity against B. cinerea. An effective combination between the antimicrobial activity of AITC-based films and the high pressure treatment was observed on a model food system (PDA) inoculated with N × 10⁴ (N ~ 1–9) conidia of B. cinerea. An HP treatment of only 300 MPa associated with an antimicrobial PLA/β-CD system providing an initial quantity of active agents equivalent to 4 mg of AITC/L of air (i.e. almost 2 folds lower than the minimal inhibition concentration of the active packaging used alone, which was determined to be equal to 10 mg/L of air in the same conditions) was found more efficient (total inhibition of B. cinerea growth during 10 days) than an 800 MPa HP treatment used alone (increase of the lag phase growth of 3.3 days).

Industrial relevance

The consumer demand for “fresh like” product containing reduced amount of preservatives without compromising human and environmental safety needs the development of new preservation strategies. As a consequence, the concept of “hurdle technologies” has risen up. The combined effect of HP treatment and volatile antimicrobial packaging allowed the use of lower individual treatment intensities to inhibit B. cinerea growth. Combining such “hurdles” is of relevance in the context of development of low-cost and eco-friendly food technologies.     

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.     

Characterization and antimicrobial analysis of chitosan-based films

Chitosan-based films for food packaging applications were prepared by casting and dried at room temperature or heat-treated in order to study functional properties and antimicrobial activity. In all cases, films were flexible and transparent, regardless of chitosan molecular weight, glycerol content, and temperature. Regarding antimicrobial activity, chitosan film forming solutions showed antimicrobial behaviour against Escherichia coli and Lactobacillus plantarum. It was also observed that the bacteriostatic property of chitosan-based films against bacteria employed in this study was notably affected by temperature. Moreover, temperature produced significant variation in the functional properties of chitosan-based films, such as colour, wettability, resistance against UV light and mechanical properties. In good agreement with this behaviour, total soluble matter (TSM), fourier transform infrared (FTIR) spectroscopy, thermo-gravimetric analysis (TGA) and X-ray diffraction (XRD) results suggested a change in the chemical structure of chitosan films, possibly due to Maillard reaction when heat treatment was used.     

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.     

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.     

Antioxidant, antifungal and antiviral activities of chitosan from the larvae of housefly, Musca domestica L.

Antioxidant activity of the chitosan from the larvae of Musca domestica L. was evaluated in two different reactive oxygen species assays, and inhibitory effects against seven fungi were also tested. The results showed that the chitosan had scavenging activity for hydroxyl and superoxide radicals which were similar to that of ascorbic acid. Also the chitosan exhibited excellent antifungal activity, especially in the low concentration, it could significantly inhibit the growth of Rhizopus stolonifer. Besides, antiviral results demonstrated that the chitosan could effectively inhibit the infection of AcMNPV and BmNPV. These results suggested that the chitosan from the larvae of housefly could be effectively used as a natural antioxidant to protect the human body from free radicals and retard the progress of many chronic diseases. Furthermore, the chitosan with antiviral and antifungal activity might provide useful information for antiviral breeding technology of economic insect and development of plant pathological control.     

Chitosan and mint mixture: A new preservative for meat and meat products

Meat is prone to both microbial and oxidative spoilage and therefore it is desirable to use a preservative with both antioxidant and antimicrobial properties. Mint extract alone had good antioxidant activity but poor antimicrobial activity, while chitosan alone showed poor antioxidant activity with excellent antimicrobial properties. Therefore, the potential of chitosan and mint mixture (CM), as a preservative for meat and meat products, was investigated. Addition of chitosan to mint extract did not interfere with the antioxidant activity of mint. In the case of 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, the IC₅₀ value for CM (17.8 μg/ml) was significantly (p ? 0.05) lower than that for mint extract (23.6 μg/ml). CM efficiently scavenged superoxide and hydroxyl radicals. The antimicrobial activities of CM and chitosan were comparable against the common food spoilage and pathogenic bacteria, the minimum inhibitory concentration being 0.05%. CM was more effective against Gram-positive bacteria. The shelf life of pork cocktail salami, as determined by total bacterial count and oxidative rancidity, was enhanced in CM-treated samples stored at 0–3 °C.     

Chitosan coatings enriched with essential oils: Effects on fungi involved in fruit decay and mechanisms of action

BackgroundPesticides have a negative impact on the health of consumers and on the environment, and as a consequence, the use of naturally occurring antimicrobial agents have become more popular to prevent food spoilage. An alternative is the use of chitosan, because of its antimicrobial, antioxidant activities and its capacity to form films having good mechanical properties. Chitosan has been used in the preparation of films and coatings for the preservation of different food products, and the addition of essential oils have been shown to be a good strategy to improve its antimicrobial activity in situ.Scope and approachThis review compiles information related to studies and research that employ chitosan as matrix for films, coatings and nanogels with essential oils extracted from plants, as antifungal agent. A special emphasis has been addressed to the effect exerted on the most pathogenic fungi for crops, also highlighting the mechanisms of action that researchers attribute to the composite, together with inhibition data.Key Findings and ConclusionEffective natural treatments, using chitosan-essential oils films or nano-emulsions, improve the preservation of fruit in terms of fungal decay. The effect is particularly dependent on the ability to release the antimicrobial compounds from the polymer matrix, the type of essential oils, the fungal species, and the incubation temperature. Several efforts are still required to understand in detail the mechanism of degradation of perishable food, how the chitosan-essential oils composites control or inhibit food decay, the effects on other pathogenic and non-pathogenic moulds as well as on the production of mycotoxins.     

Physical and antimicrobial properties of banana flour/chitosan biodegradable and self sealing films used for preserving Fresh-cut vegetables

Kluai Namwa is a local species of banana grown in Thailand. Flour made from this banana was mixed with chitosan, a by product of the shrimp industry, for casting films as it has valuable characteristics including antimicrobial effects. Utilization of excess banana available in high season and chitosan can from the waste crustacean shells could help reduce waste while making available a value added product. Banana/chitosan films were produced using 0.5–2 g banana flour and 0.5 g chitosan in 100 ml aqueous solution. Film water vapor permeability, tensile properties, solubility and morphology were investigated. The composite yellowish film exhibited great water permeability of 38.81–41.66 g mm/m² day kPa. Tensile strength and elongation were in the range of 5.19–14.22 MPa and 1.64–2.59%, respectively while the solubility obtained was 40.90–64.21%. The presence of starch in the composite film makes possible water soluble and sealable bags or wraps, while the presence of chitosan gives them the antimicrobial property. The composite bags were found to protect asparagus, baby corn and Chinese cabbage against Staphylococcus aureus activity by serving as a good barrier and as a antimicrobial agent. With these properties the edible bag with its contents can be processed together during food preparation making its use very convenient.     

Preparation,characterization and antimicrobial study of a hydrogel (soft contact lens) material impregnated with silver nanoparticles

PurposeContact lenses that incorporate antimicrobial properties may reduce the risk for microbial-associated adverse events for lens wearers. The aim of this study was to assess the antimicrobial effects of silver nanoparticles (NP) when impregnated in a hydrogel material.MethodsHydrogel disks, used as a proxy for soft contact lenses, were prepared with silver NPs to add an antimicrobial effect to the polymer. Six groups of disks were created, each with a different concentration of silver NPs. The antimicrobial effect of the hydrogels against Pseudomonas aeruginosa (ATCC15442) and Staphylococcus aureus (ATCC6538) was evaluated at 6, 24, 48 and 72 h.ResultsSilver NP concentrations ranged from 20.71 to 98.06 μg/disk. All groups demonstrated excellent antibacterial effects against P. aeruginosa at each time point. After 6 h all disks didn’t exhibit desirable antibacterial activity against S. aureus; whereas except those with 20.71 μg silver NPs showed antibacterial activity at 24 h and only the disks with 57.13 and 98.06 μg silver NPs showed antimicrobial activity at 48 and 72 h.ConclusionsThe development of contact lenses made of a silver NP-impregnated hydrogel material may bring antimicrobial effects sufficient to decrease the risk of microbial-related adverse events for lens wearers.     

Chitosan Controls Postharvest Decay and Elicits Defense Response in Kiwifruit

Chitosan is an eco-friendly alternative to synthetic fungicides for managing postharvest decay of fruits and vegetables. The ability of chitosan to enhance resistance to postharvest gray (Botrytis cinerea) and blue mold (Penicillium expansum) in kiwifruit was investigated. The gene expression and activity of antioxidant enzymes, and total phenolic compounds were determined. Results showed that chitosan at 5 g/L significantly inhibited gray and blue mold in kiwifruit stored at 25 and 4 °C. Chitosan significantly induced the gene expression of catalase, superoxide dismutase, and ascorbate peroxidase, as well as increased the enzyme activity. Moreover, chitosan markedly increased the content of total phenolic compounds in kiwifruit. Importantly, chitosan also exhibited beneficial effects on fruit quality. Taken together, the ability of chitosan to reduce mold in stored kiwifruit may be associated with the elicitation of host defense response. These results have practical implications for the application of chitosan to reduce postharvest losses.     

Botryticidal Activity of Nanosized Silver‐Chitosan Composite and Its Application for the Control of Gray Mold in Strawberry

Botrytis cinerea Pers, the gray mold fungus, is among the most dangerous plant pathogens that cause great losses in agricultural crops. The botryticidal activities of nanosized silver (nano Ag), fungal chitosan (CTS) irradiated fungal chitosan (IrCTS), and nano Ag‐IrCTS composite were investigated. All of the examined materials exhibited powerful antifungal activity against B. cinerea. The most effective compound was nano Ag‐IrCTS composite with a minimal inhibitory concentration of 125 μg/mL. The microscopic examination, of treated B. cinerea with the composite, revealed that an obvious alteration in mycelial shape was appeared accompanied and moderate lysis in fungal hyphae. With the prolongation of treatment, most of the fungal mycelia were lysed into small and elastic fragments. The consequence of strawberries coating, with antifungal composite based solution, was the disappearance of gray mold infection signs in 90% of the contaminated fruits after 7 d of storage, treated fruits had a fresh‐like appearance at the end of storage. Coating with nano Ag‐IrCTS solution could be highly recommended regarding its efficiency in prohibiting B. cinerea growth, preventing gray mold decay and enhancing the overall quality of coated strawberry fruits.     

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.     

In vivo and in vitro antifungal effect of fungal chitosan nanocomposite edible coating against strawberry phytopathogenic fungi

This study compared the effectiveness of fungal chitosan nanocomposite, chitosan gel and chitosan nanoparticles against strawberry phytopathogenic fungi (Botrytis cinerea; Rhizopus stolonifer and Aspergillus niger). Nanoparticles were prepared by ionic gelation method and characterised by dynamic light scattering and scanning electron microscopy. The influence of the edible coatings on fungal growth was analysed in vitro and in vivo The fungal chitosan nanoparticles presented an average size = 331.1 nm (±7.21) with a narrow size distribution (PDI = 0.377) and a zeta potential = +34 mV. The edible coating made by the nanocomposite exhibited important changes in fungal morphology, and the best control of the growth of the assayed fungal strains in artificially infected strawberries. Therefore, nanotechnology brought some benefits to the conventional chitosan gel edible coating, improving its antifungal activity and forming a new eco-friendly coating.     

Stabilized nanosilver based antimicrobial poly(3-hydroxybutyrate-co-3-hydroxyvalerate) nanocomposites of interest in active food packaging

Antimicrobial silver based nanocomposites of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) were successfully synthesized and characterized. For the synthesis, a masterbatch of in situ stabilized silver nanoparticles (AgNPs) produced into a mixed microbial cultures based poly(3-hydroxybutyrate-co-18 mol%-3-hydroxyvalerate) (PHBV18) was used, which was diluted by melt compounding with a commercial poly(3-hydroxybutyrate-co-3 mol%-3-hydroxyvalerate) (PHBV3) material. The incorporated AgNPs (0.04 wt.%) led to a surprising oxygen permeability drop of ca. 56% compared to the neat polymer. The thermal stability and optical properties of the nanocomposites were not significantly modified as compared to the neat PHBV3. Moreover, the antimicrobial performance of the PHBVs-AgNPs films against two of the most common food borne pathogens, Salmonella enterica and Listeria monocytogenes, showed a strong and sustained (even after seven-months) antibacterial activity. This study provides an innovative route to generate fully renewable and biodegradable antimicrobial nanocomposites that could potentially be of interest in film and coating applications such as active food packaging.Industrial relevanceAs a response to the consumers for more safety foodstuffs and ecofriendly packaging materials, this work presents a novel methodology to develop antimicrobial packaging by using biodegradable materials obtained from industrial food by-products in combination of an industrially meaningful melt blending process. The methodology here applied allows the use of low doses of stabilized silver nanoparticles in the polymer matrix, without additives, which exhibits prolonged antimicrobial activity against food borne pathogens and enhanced oxygen barrier properties. These materials are of great interest in the development and design of biodegradable active food packaging materials and antibacterial food contact surfaces with the additional advantage that they can be easily scale-up.     

Application of chitosan-incorporated LDPE film to sliced fresh red meats for shelf life extension

Chitosan lactate was impregnated as an antimicrobial additive into low density polyethylene (LDPE) with different concentrations. The antimicrobial effectiveness was tested with three pathogenic bacteria, specifically Listeria monocytogenes, Escherichia coli and Salmonella enteritidis. Also, these chitosan incorporated films were applied on red meat surfaces to determine the effectiveness of chitosan on color shelf life extension and microbial growth inhibition. Chitosan was exposed to 0.1% peptone water containing the three pathogens in separate tests and inhibited microbial growth a higher levels with increasing concentration of chitosan in the film matrix. Oxygen permeability was not affected by the incorporation of chitosan, while the water vapor permeability increased with the addition of chitosan. Film elongation decreased with the addition of chitosan. When chitosan incorporated films were applied on fresh red meat, microorganisms on the meat surface were not inhibited but significant extension of red color shelf life were observed in refrigerated, sliced red meats.     

Characterisation and cooperative antimicrobial properties of chitosan/nano-ZnO composite nanofibrous membranes

Chitosan was combined with nano-ZnO to increase its antimicrobial activity, using polyvinyl alcohol as a support, and then were electronspun to form composite nanofibres. Through SEM, EDX and XRD observations, chitosan was seen to be able to incorporate nano-ZnO in the composite nanofibres. Escherichia coli, expressing recombinant enhanced green fluorescent protein, and Candida albicans were used to test the antimicrobial efficacy of the newly synthesised chitosan/nano-ZnO antimicrobial composite. The CdTe quantum dots were used to rapidly detect the residual changes of C. albicans and determine the end point of using antimicrobial agents. Minimal minimum inhibitory concentration (MIC), post-antibiotic effect and continuous agent effect of the composite were determined. The MIC of chitosan/nano-ZnO against C. albicans was 160 μg/ml, close to the concentration of the treated composite with the lowest fluorescence intensity. The cell damage was observed by SEM, which indicated that nano-ZnO in the nanofibrous membranes played a cooperative role in the antimicrobial process of chitosan.