Effect of Biodegradable Coatings on the Growth of Aspergillus flavus In Vitro, on Maize Grains, and on the Quality of Tortillas during Storage.

The fungus Aspergillus flavus causes serious damage to maize grains and its by-products, such as tortilla. Currently, animal and plant derivatives, such as chitosan and propolis, and plant extract residues, respectively, are employed as alternatives of synthetic fungicides. The objective of this research was to evaluate the efficacy of several formulations based on propolis-chitosan-pine resin extract on the in vitro growth of A. flavus, the growth of maize grain plantlets and the quality of stored tortillas at 4 and 28 °C. The most outstanding formulation was that based on 59.7% chitosan + 20% propolis nanoparticles + 20% pine resin extract nanoparticles; since the in vitro conidia germination of A. flavus did not occur, disease incidence on grains was 25-30% and in tortillas, 0% infection was recorded, along with low aflatoxin production (1.0 ppb). The grain germination and seedling growth were markedly reduced by the nanocoating application. The percentage weight loss and color of tortillas were more affected by this coating compared to the control, and the rollability fell within the scale of non-ruptured at 4 °C and partially ruptured at 28 °C. The next step is to evaluate the toxicity of this formulation.

Non-Chemical Treatments for the Pre- and Post-Harvest Elicitation of Defense Mechanisms in the Fungi-Avocado Pathosystem.

The greatest challenge for the avocado (Persea americana Miller) industry is to maintain the quality of the fruit to meet consumer requirements. Anthracnose is considered the most important disease in this industry, and it is caused by different species of the genus Colletotrichum, although other pathogens can be equally important. The defense mechanisms that fruit naturally uses can be triggered in response to the attack of pathogenic microorganisms and also by the application of exogenous elicitors in the form of GRAS compounds. The elicitors are recognized by receptors called PRRs, which are proteins located on the avocado fruit cell surface that have high affinity and specificity for PAMPs, MAMPs, and DAMPs. The activation of defense-signaling pathways depends on ethylene, salicylic, and jasmonic acids, and it occurs hours or days after PTI activation. These defense mechanisms aim to drive the pathogen to death. The application of essential oils, antagonists, volatile compounds, chitosan and silicon has been documented in vitro and on avocado fruit, showing some of them to have elicitor and fungicidal effects that are reflected in the postharvest quality of the fruit and a lower incidence of diseases. The main focus of these studies has been on anthracnose diseases. This review presents the most relevant advances in the use of natural compounds with antifungal and elicitor effects in plant tissues.

Effective In Vitro Control of Two Phytopathogens of Agricultural Interest Using Cell-Free Extracts of Pseudomonas fluorescens and Chitosan.

A biofungicide is a natural product that can be derived from various sources such as, among others, microorganisms, higher plants, animal products, phytochemicals, semiochemicals, and antagonist microorganisms. One of the most important approaches for the production of biofungicides is the combination of biocontrol agents. This study showed the inhibition growth of Alternaria alternata and Fusarium solani treated with cell-free extracts of P. fluorescens. Using thin-layer chromatography and plate assays it was also demonstrated that the cell-free extracts of P. fluorescens contained siderophores and derivates of 4-diacetylphloroglucinol and phenazine. Moreover, the combination of cell-free extracts of P. fluorescens and chitosan [50-1.5% (v/v)] had a synergistic effect since they notably inhibited the mycelial growth of A. altenata and F. solani. Various morphological alterations to the mycelia and conidia of the treated fungi as a result of this combination were also observed. The present study could be a starting point to control other fungal phytopathogens using different cell-free extracts and chitosan as biocontrol agents.

Physical-Chemical Evaluation of Active Food Packaging Material Based on Thermoplastic Starch Loaded with Grape cane Extract.

The aim of this paper is to evaluate the physicochemical and microbiological properties of active thermoplastic starch-based materials. The extract obtained from grape cane waste was used as a source of stilbene bioactive components to enhance the functional properties of thermoplastic starch (TPS). The biomaterials were prepared by the compression molding technique and subjected to mechanical, thermal, antioxidant, and microbiological tests. The results showed that the addition of grape cane extract up to 15 wt% (TPS/WE15) did not significantly influence the thermal stability of obtained biomaterials, whereas mechanical resistance decreased. On the other side, among all tested pathogens, thermoplastic starch based materials showed antifungal activity toward Botrytis cinerea and antimicrobial activity toward Staphylococcus aureus, suggesting potential application in food packaging as an active biomaterial layer.

Cellular alterations in Pectobacterium carotovorum treated with nanostructured formulations during the incubation time.

Pectobacterium carotovorum was incubated in formulations of chitosan nanoparticles or thyme essential oil-loaded chitosan nanoparticles for a maximum period of 48 h time. The cellular changes and viability were evaluated by transmission electron microscopy (TEM), and two colorimetric assays 3-(4,5 dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide (MTT) and alamar blue (AMB), respectively. The incubation time and the addition of the secondary metabolite to the formulation were key factors to the cell damage and cell inhibitory effects on P. carotovorum, TEM observations overall demonstrated on the treated bacterium, cell surface alterations such as deforming and disappearance of the cell wall and the plasmatic membrane, with agglomeration of nanoparticles outside and inside of the cells, loss of cell content and lysis. Cell viability was reduced about 80% (MTT) and 100% (AMB) in the applied treatment of chitosan-loaded thyme essential oil nanoparticles after 48 h incubation, in addition, the total cell inhibition was shown from 6 h incubation onwards with the AMB assay. The TEM micrographs and the cell viability assays provided enough evidence of the antimicrobial activity of the nanostructured formulations compared with the control where no damage was observed.

Cactus stem (Opuntia ficus-indica Mill): anatomy, physiology and chemical composition with emphasis on its biofunctional properties.

Cactus stem (Opuntia ficus-indica Mill) is native to Mesoamerica and marketed in different forms such as fresh, frozen or pre-cooked. Worldwide, this vegetable is recognized for its pharmaceutical actions, including its antioxidant, diuretic, anticarcinogenic, anti-inflammatory, anti-diabetic, and anti-hypercholesterolemic properties, as well as their antiviral and antispermatogenic effects. However, not all of these properties have been associated with its chemical composition; therefore, this review aims to present and integrate information available on the physiology and anatomy of cactus stem and its chemical composition, focusing on some of the many factors that determine its biofunctionality. © 2017 Society of Chemical Industry.

The Response of Naturally Based Coatings and Citrus Fungicides to the Development of Four Postharvest Fungi.

The tomato (Licopersicon esculentum Mill.) is considered to be one of the products with the highest demand due to its nutritional value; however, it is susceptible to infection by fungi during its pre- and postharvest stages. In this research, three commercial products (1% Citrocover, 1% Citro 80, and 0.002% Microdyn) and two coatings based on 1.0% chitosan/0.1% lime or 0.1% orange essential oils were evaluated in vitro and on Saladette tomatoes that were previously inoculated with four postharvest fungi. The application of the commercial citrus-based product Citrocover was highly effective in reducing the in vitro development of Aspergillus flavus, Fusarium oxysporum, and Colletotrichum gloeosporioides, but not Rhizopus stolonifer. The sanitizer Microdyn promoted infections with most fungi. Citrus-based products were effective in reducing infections with A. flavus in the tomatoes during storage. Overall, mycotoxin production was very low for all treatments. The use of commercial citrus-based products and coatings did not alter the weight loss, firmness, or total soluble solid contents of the treated tomatoes. The changes observed were, rather, associated with the normal ripening process of Saladette tomatoes. The commercial citrus-based products satisfactorily controlled the in vitro growth of the fungi Aspergillus flavus, Fusarium oxysporum, and Colletotrichum gloeosporioides.

Biodegradability Assessment of Prickly Pear Waste-Polymer Fibers under Soil Composting.

Nowadays, solving the problems associated with environmental pollution is of special interest. Therefore, in this work, the morphology and thermal and mechanical properties of extruded fibers based on polylactic acid (PLA) and poly(butylene adipate-co-terephthalate) (PBAT) added to prickly pear flour (PPF) under composting for 3 and 6 months were evaluated. The highest weight loss percentage (92 ± 7%) was obtained after 6-month degradation of the PLA/PBAT/PPF/CO/AA blend, in which PPF, canola oil (CO), and adipic acid (AA) were added. Optical and scanning electron microscopy (SEM) revealed structural changes in the fibers as composting time increased. The main changes in the absorption bands observed by Fourier transform infrared spectroscopy (FTIR) were related to the decrease in -C=O (1740 cm-1) and -C-O (1100 cm-1) groups and at 1269 cm-1, associated with hemicellulose in the blends with PPF. Differential scanning calorimetry (DSC) showed an increase in the cold crystallization and melting point with degradation time, being more evident in the fibers with PPF, as well as a decrease in the mechanical properties, especially Young's modulus. The obtained results suggest that PPF residues could promote the biodegradability of PLA/PBAT-based fiber composites.

The Effect of Netting Bags on the Postharvest Quality, Bioactive and Nutritional Compounds, and the Spoilage Microorganisms Content of Bell Peppers.

The packaging nets used for bell peppers act as a form of protection. However, the manufacturing is based on polymers that cause serious environmental problems. The effects of nets made of biodegradable materials, such as poly(lactic) acid (PLA), poly(butylene adipate-co-terephthalate) (PBAT), and cactus stem residues, were evaluated on four different colors of 'California Wonder' bell peppers stored over a 25-day storage period under controlled and ambient temperature conditions. Compared to commercial polyethylene nets, the bell peppers kept in the biodegradable nets did not show notable differences with respect to color, weight loss, total soluble solids, and titratable acidity. However, there were significant differences (p < 0.05) in terms of phenol content, carotenoids (orange bell peppers), anthocyanins, and vitamin C, with an overall tendency to show a higher content in those kept in PLA 60%/PBTA 40%/cactus stem flour 3% compared to commercial packaging. In addition, the same net notably reduced the development of bacteria, fungi, and yeasts during the storage of red, orange, and yellow bell peppers. As postharvest packaging for bell peppers, this net could be considered a viable option for the storage of this product.

Anthracnose Disease Control and Postharvest Quality of Hass Avocado Stored in Biobased PLA/PBAT/Pine Essential Oil/Chitosan Active Packaging Nets.

For sale in the domestic market, Hass avocados are kept in non-biodegradable plastic nets and stored at an ambient temperature. The fungus Colletotrichum gloeosporioides can cause important losses at this stage. Consequently, formulations based on polylactic acid (PLA)/poly (butylene adipate-co-terephthalate) (PBAT) 60/40 biodegradable blends added with pine essential oil (PEO) at 10, 12, 14, and 20% and coated with 1% chitosan (CH) were used for the elaboration of nets, which were evaluated on C. gloeosporioides and Rhizopus stolonifer on nutrient media and fruit and on the ripening behavior of Hass avocados at ambient temperature. The spore germination stage of C. gloeosporioides was the most extensively damaged (78% inhibition). The incidence of anthracnose was notably reduced by almost 80% in the avocados stored in the 60/40 PLA/PBAT nets coated with 1% CH. The overall values regarding weight loss, °Brix, and dry matter were c.a. 23.5%, 7.5, and 24.5%, respectively. The changes in firmness, color, and CO2 production were particularly associated with the initial harvest index and storage temperature rather than with the nets. In future research, essays on nets should include evaluations at the commercial levels.

Detection of Alternaria alternata in tomato juice and fresh fruit by the production of its biomass, respiration, and volatile compounds.

Tomato is widely consumed and marketed as juice, puree, or fresh product. Nevertheless, 30% of its harvest volume is lost because of the fungus Alternaria alternata. This research aimed to provide early detection methods for this fungal decay on tomato juice and fresh fruit. Biomass content, CO2, O2 and volatile compounds (VOCs) during A. alternata growth in tomato juice and fruit at two ripening stages (breaker and red colour) were evaluated. Additionally, CO2 and VOCs data set were analysed with a hierarchical cluster technique (HCA) to explore the differences between inoculated and non-inoculated samples. Biomass was determined by gravimetry, CO2 and O2 by gas chromatography (GC), and VOCs by GC-mass spectrometry. Biomass content was not drastically modified by tomato's ripening stage (3-6 mg of dry weight). CO2 in tomato juice was considerably higher in the inoculated samples with A. alternata (27-63%) than in the non-inoculated ones (2.8-6.6%), regardless of the ripeness stage; while in tomato fruit CO2 was higher at breaker stage and inoculated with A. alternata (33-41%) than the remaining treatments (9-23%). It was also observed that, except for limonene, trans-sabinene hydrate, and rhodovibrin, VOCs' release during the interaction between tomato juice and A. alternata was different from the fresh tomato and A. alternata interaction. Only the HCA based on CO2 data showed clear differences between the inoculated and non-inoculated tomato juice and fruit at both ripening stages.

Application of natural-based nanocoatings for extending the shelf life of green bell pepper fruit.

Pectobacterium carotovorum is a phytopathogenic bacteria that causes significant economic loses in food crops, such as bell pepper, which is of special significance in the value of production and trade in Mexico. Therefore, a solution for fruit conservation must be sought. Due to environmental concerns, it is necessary the use of environmentally-friendly active packaging. In this article, chitosan and chitosan-thyme essential oil nanocoatings were used for the preservation of green bell pepper. Different formulations based on chitosan nanoparticles (CSNPs) and chitosan-thyme essential oil nanoparticles (15, 30, and 45%) were prepared. For uncoated and coated bell peppers, the quality and physiological variables of inoculated and uninoculated fruit with P. carotovorum during 12-day storage period were assessed. According to the results, the weight loss of the fruit remained almost constant over the storage days for the different formulations. A decrease in fruit firmness and an increase in the respiration rate and ascorbic acid content until day 8 with a decrease at the end of the storage period were observed. Of all the evaluated nanocoatings, the fruit treated with the formulation containing 15% CSNPs showed the lowest colony-forming units and disease incidence. Also, the coated bell peppers with this formulation had lower CO2 production compared to the remaining treatments, and the weight loss and firmness were maintained. Therefore, the use of CSNP coatings could represent a good alternative for the protection of bell pepper against the pathogenic bacteria P. carotovorum. PRACTICAL APPLICATION: The results of the application of nanocoatings based on chitosan and chitosan-thyme essential oil as an antibacterial agent against P. carotovorum on green bell pepper during 12-day storage period suggest that nanoparticle-based coatings can be a natural option for the preservation of fruit quality during ripening.

Use of Natural Products on the Control of Aspergillus flavus and Production of Aflatoxins In Vitro and on Tomato Fruit.

Aspergillus flavus affects fresh and dry fruit and vegetable products, and its toxic metabolites, namely aflatoxins, cause serious damage in humans. The objective of this research study was to evaluate the effect of commercial natural products as well as edible and nanostructured chitosan coatings on the development of A. flavus and on the production of aflatoxins in vitro and in tomato. Treatments were as follows: chitosan 1%, chitosan coating, chitosan nanostructured coating, Citrocover 1% (citrus seed extract), Resinadher 0.5% (pine resin extract), mancozeb 2%, and water. The variables were as follows: halo inhibition, spore production, and aflatoxins content. In fruit, the following were evaluated: disease incidence, mycelial growth, and aflatoxin production. An ANOVA (Tukey: p < 0.05) was used. In vitro results showed that Citrocover and Resinadher reduced sporulation (0.2 and 0.9 × 105 spores mL-1, respectively), while chitosan inhibited the production of aflatoxins. With Resinadher and Citrocover, tomato fruit had the lowest incidence, mycelial growth, and aflatoxin production with corresponding values of 0%, 0.0 cm2, and 0.95 ppb, respectively, and 7%, 0.2 cm2, and 1.77 ppb, respectively. The use of Citrocover and Resinadher could be a viable alternative to decrease the development of A. flavus in tomato fruit.

Monitoring the infection process of Rhizopus stolonifer on strawberry fruit during storage using films based on chitosan/polyvinyl alcohol/polyvinylpyrrolidone and plant extracts.

Different formulations based on nanoparticles of chitosan-plant extracts were evaluated to detect the infection process from the earliest stage of the fungus Rhizopus stolonifer on strawberry fruit during storage. Chitosan/polyvinyl alcohol (Ch/PVA) and chitosan/polyvinylpyrrolidone (Ch/PVP) films enriched with nanoparticles (NPs) of chitosan blended with plant extracts were prepared. They were placed inside a plastic package containing inoculated fruits and stored at 25 °C for 72 h. The thickness values of the films were in the range of 0.10 to 0.25 mm. All samples showed a maximum absorbance peak of about 300-320 nm; however, the Ch/PVP films enriched with NPs of chitosan and 10% of radish extract had an evident decrease in the optical absorbance as the fungal infection progressed. Additionally, as observed by scanning electron microscopy, the cross-section and surface morphology of films were not modified during storage, and the growth of R. stolonifer was evident after 48 h. Therefore, the Ch/PVP films enriched with chitosan NPs blended with 10% radish extract could be a reliable indicator of this fungus's growth.

Edible Chitosan/Propolis Coatings and Their Effect on Ripening, Development of Aspergillus flavus, and Sensory Quality in Fig Fruit, during Controlled Storage.

Biodegradable alternatives for the control of Aspergillus flavus in fig fruit were tested with the application of coatings based on chitosan (CS) and propolis (P). To potentiate the fungicidal effect, nanoparticles of these two (CSNPs and PNPs) were also considered. The objectives of this research were to evaluate the effect of different formulations on: (a) the ripening process of the fig, (b) the incidence of A. flavus and the production of aflatoxins, and (c) the acceptance of the treated fruit by a panel. The nanostructured coatings did not influence the ripening process of the fruit during the 12 days of storage, however, the antioxidant activity increased by approximately 30% with the coating CS + PNPs + P. The figs treated with CS + CSNPs + PNPs + P, inhibited the growth of the fungus by about 20% to 30% under laboratory and semi-commercial conditions. For all treatments, the aflatoxin production was lower than 20 ppb compared to the control with values of c.a. 250 ppb. The sensory quality was acceptable among the panel. The edible coatings can be a non-toxic alternative for post-harvest preservation and the consumption of fig fruit. The next step will be its inclusion and evaluation at a commercial level in packing houses.

Corn-Starch-Based Materials Incorporated with Cinnamon Oil Emulsion: Physico-Chemical Characterization and Biological Activity.

Active packaging represents a large and diverse group of materials, with its main role being to prolong the shelf-life of food products. In this work, active biomaterials based on thermoplastic starch-containing cinnamon oil emulsions were prepared by the compression molding technique. The thermal, mechanical, and antifungal properties of obtained materials were evaluated. The results showed that the encapsulation of cinnamon oil emulsions did not influence the thermal stability of materials. Mechanical resistance to break was reduced by 27.4%, while elongation at break was increased by 44.0% by the addition of cinnamon oil emulsion. Moreover, the novel material provided a decrease in the growth rate of Botrytis cinerea by 66%, suggesting potential application in food packaging as an active biomaterial layer to hinder further contamination of fruits during the storage and transport period.

Nanostructured chitosan edible coating loaded with α-pinene for the preservation of the postharvest quality of Capsicum annuum L. and Alternaria alternata control.

Bell peppers are susceptible to postharvest diseases caused by the fungus Alternaria alternata that limit its commercialization. Nowadays, nanotechnology allows encapsulation of natural components such as terpenes. The objective of this work was to develop chitosan nanoparticles with α-pinene (P-CSNPs) and a nanostructured edible coating (EC-P-CSNPs). The P-CSNPs were characterized by TEM (Transmission Electron Microscopy), FTIR (Fourier-Transform Infrared Spectroscopy), DLS (Dynamic Light Scattering) and ζ potential. The P-CSNPs and the EC-P-CSNPs were applied to the bell peppers inoculated with A. alternata under cold storage for either 0, 7, 14 and 21 days at 12 ±â€¯2 °C followed by a shelf-life period of 5 days at 20 ±â€¯2 °C to assess their post-harvest quality. Nanoparticles size was 3.9 ±â€¯0.5 nm and the ζ potential value was between 13.4 and 14.9 mV. The incorporation of α-pinene was corroborated by FTIR. Significant changes in weight loss were obtained for P-CSNPs and EC-P-CSNPs at percentage of 3 and 6% compared to the control. For firmness, color, total soluble solids, titratable acids, maturity index, total flavonoid content and antioxidant capacity, no differences were found. Total carotenes were higher in bell peppers without A. alternata. The chitosan nanoparticles and edible coating inhibited A. alternata during the cold storage period of bell pepper and preserved the physicochemical quality.

Nanostructured chitosan/propolis formulations: characterization and effect on the growth of Aspergillus flavus and production of aflatoxins.

A great diversity of agricultural products is susceptible to contamination caused by Aspergillus flavus. To reduce fungal contamination, the application of natural products has been proposed, including chitosan and propolis, due to its broad and recognized antimicrobial activity on several microorganisms. Currently, the application of nanotechnology allows for a greater activity to be more reactive and efficient. The objectives of this research were to characterize by TEM and Z potential some of the studied nanoparticles and to determine the in vitro antifungal activity of the formulations and the production of aflatoxins of the treated fungus. For this, individual treatments and different nanoformulations were elaborated by varying the percentage of the components such as chitosan solution, chitosan nanoparticles, an extract of propolis, nanoparticles of propolis, glycerol and canola oil. The final concentrations of the formulations were of 20%, 30% and 40% and the control consisted of Czapeck-dox agar medium. TEM micrographies showed a spherical morphology in a range of 2.3-3.0 nm with values of Z potential from 18.5 to 116.2 nm. Compared to the untreated fungus, the highest effect was seen in the parameter of spore germination, since inhibition was of c. a. 97% corresponding to the formulation containing chitosan + propolis nanoparticles + chitosan nanoparticles + propolis extract at the highest concentration of 40%. At this same concentration, the production of aflatoxins was 100% inhibited with the treatment with chitosan at 1%. Since these results are under carefully controlled conditions, further research should be extended to different fruit and vegetables affected by this fungus.

Estimating CO2 and VOCs production of Colletotrichum fragariae and Rhizopus stolonifer grown in cold stored strawberry fruit.

The aim of this work was to investigate the early detection of anthracnose and soft rot diseases in cold stored strawberry fruit by evaluating the CO2 and volatile organic compounds (VOCs) released by the fungi Colletotrichum fragariae and Rhizopus stolonifer. Strawberries were stored at 5, 10 and 21 °C (control group) and the VOCs and CO2 production of inoculated and non-inoculated strawberries were followed by gas chromatography. To evaluate and estimate the growth of both fungi, the CO2 data were fitted to the Gompertz model. Data of the VOCs released at the end of the fungal growth were analyzed using principal components analysis (PCA) to discriminate between infected and non-infected strawberries. The results showed that fungal growth was affected by temperature and C. fragariae had a maximum growth after 14.6 h at 5 °C and R. stolonifer at 21 °C after 45.2 h. On the other hand, through VOCs released by C. fragariae and R. stolonifer and PCA, four groups were obtained: a) strawberry infected with C. fragariae, stored at 10 °C, b) strawberry infected with R. stolonifer, stored at 21 °C, c) control group kept at 10 °C and, d) strawberry infected with C. fragariae and control group (5 and 21 °C), and strawberry infected with R. stolonifer at 5 and 10 °C. In conclusion, CO2 and VOCs released by C. fragariae and R. stolonifer on strawberries could infer the presence of anthracnose and soft rot during storage of the fruit at low temperature.

Preparation and Characterization of Bio-Based PLA/PBAT and Cinnamon Essential Oil Polymer Fibers and Life-Cycle Assessment from Hydrolytic Degradation.

Nowadays, the need to reduce the dependence on fuel products and to achieve a sustainable development is of special importance due to environmental concerns. Therefore, new alternatives must be sought. In this work, extruded fibers from poly (lactic acid) (PLA) and poly (butylene adipate-co-terephthalate) (PBAT) added with cinnamon essential oil (CEO) were prepared and characterized, and the hydrolytic degradation was assessed. A two-phase system was observed with spherical particles of PBAT embedded in the PLA matrix. The thermal analysis showed partial miscibility between PLA and PBAT. Mechanically, Young's modulus decreased and the elongation at break increased with the incorporation of PBAT and CEO into the blends. The variation in weight loss for the fibers was below 5% during the period of hydrolytic degradation studied with the most important changes at 37 °C and pH 8.50. From microscopy, the formation of cracks in the fiber surface was evidenced, especially for PLA fibers in alkaline medium at 37 °C. This study shows the importance of the variables that influence the performance of polyester-cinnamon essential oil-based fibers in agro-industrial applications for horticultural product preservation.