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.

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.

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.