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.     

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.     

Effect of salep‐based edible coating enriched with grape seed extract on postharvest shelf life of fresh strawberries

Effects of novel edible coatings based on salep solution (SS) enriched with grape seed extract (GSE) (1.5% SS, 1.5% SS + 0.5% GSE, 1.5% SS + 1.5% GSE, 1.5% SS + 3% GSE) on the quality characteristics of fresh strawberries were monitored after 0, 4, 8, 12, 16, and 20 days of storage (1°C, 95% RH). The lowest growth of microorganisms, weight loss, softening and ascorbic acid, phenol and anthocyanin degradation was measured in strawberries coated with 1.5% SS + 3% GSE. The highest superoxide dismutase activity and the lowest peroxidase activity were determined in strawberries coated with 1.5% SS + 3% GSE. Moreover, they received the highest ranked sensory attributes. Incorporating GSE into the SS coating formulation as a new hurdle increased its antimicrobial properties and extended the shelf life of coated strawberries compared to uncoated by up to 20 days.     

Antifungal effectiveness of potassium sorbate incorporated in edible coatings against spoilage molds of apples, cucumbers, and tomatoes during refrigerated storage

Abstract: Predominant spoilage molds of fresh apples, cucumbers, and tomatoes stored at 4 °C were isolated and examined for resistance to potassium sorbate (PS) incorporated in polysaccharide edible coatings. The isolated molds were Penicillium expansum, Cladosporium herbarum, and Aspergillus niger from apples. P. oxalicum and C. cucumerinum were isolated from cucumbers and P. expansium and C. fulvum from tomatoes. Guar gum edible coating incorporated with PS was the most effective mold inhibitor, significantly (P < 0.05) reducing the isolated spoilage molds for 20, 15, and 20 d of storage at 4 °C on apples, cucumbers, and tomatoes, respectively. PS incorporated into pea starch edible coating was less effective and selectively inhibited the isolated mold species, causing significant (P < 0.05) reduction in mold on apples, cucumbers, and tomatoes counts for 20, 10 to 15, and 15 to 20 d of storage at 4 °C, respectively. The isolated mold species exhibited different resistances to PS incorporated in the edible coatings. The greatest inhibition (2.9 log CFU/g) was obtained with C. herbarum on apples and the smallest (1.1 log CFU/g) was with P. oxalicum on cucumbers and the other isolated mold species exhibited intermediate resistance. The coatings tested, in general, inhibited molds more effectively on apples than on tomatoes and cucumbers. Addition of PS to pea starch and guar gum, edible coatings improved the antifungal activity of PS against isolated spoilage molds on apples, cucumbers, and tomatoes. PS inhibition was most effective against C. herbarum on apples and least effective against P. oxalicum on cucumbers.     

Mold growth on strawberries and cherries during storage at 25 degrees C

Mold species that grew on the surface of retailed strawberries (10 packs, 211 strawberries) and cherries (18 packs, 441 cherries) during storage at 25 degrees C were isolated and identified to evaluate the state of mold growth. Mold growth was observed on 208 (98.6%) of the 211 strawberries and 193 (43.8%) of the 441 cherries. The mold species most frequently isolated from strawberries was Botrytis cinerea, being observed in 81.0% of the strawberries examined, followed by Cladosporium and Alternaria alternata. The mold most frequently isolated from cherries was Alternaria (28.7%), followed by Penicillium, Botrytis, and Cladosporium. The frequency of cherries on which mold growth was observed varied among packs. Mold tended to grow more often in the areas of the fruits in contact with adjacent fruits.     

Mould and yeast flora in fresh berries, grapes and citrus fruits

Fresh fruits are prone to fungal contamination in the field, during harvest, transport, marketing, and with the consumer. It is important to identify fungal contaminants in fresh fruits because some moulds can grow and produce mycotoxins on these commodities while certain yeasts and moulds can cause infections or allergies. In this study, 251 fresh fruit samples including several varieties of grapes, strawberries, blueberries, raspberries, blackberries, and various citrus fruits were surface-disinfected, incubated at room temperature for up to 14 days without supplemental media, and subsequently examined for mould and yeast growth. The level of contamination (percent of contaminated items/sample) varied depending on the type of fruit. All raspberry and blackberry samples were contaminated at levels ranging from 33% to 100%, whereas 95% of the blueberry samples supported mould growth at levels between 10% and 100% of the tested berries, and 97% of strawberry samples showed fungal growth on 33-100% of tested berries. The most common moulds isolated from these commodities were Botrytis cinerea, Rhizopus (in strawberries), Alternaria, Penicillium, Cladosporium and Fusarium followed by yeasts, Trichoderma and Aureobasidium. Thirty-five percent of the grape samples tested were contaminated and supported fungal growth; the levels of contamination ranged from 9% to 80%. The most common fungi spoiling grapes were Alternaria, B. cinerea and Cladosporium. Eighty-three percent of the citrus fruit samples showed fungal growth at levels ranging from 25% to 100% of tested fruits. The most common fungi in citrus fruits were Alternaria, Cladosporium, Penicillium, Fusarium and yeasts. Less common were Trichoderma, Geotrichum and Rhizopus.     

Effect of chitosan coating combined with postharvest calcium treatment on strawberry (Fragaria × ananassa) quality during refrigerated storage

Strawberries (Fragaria × ananassa Duch.) were coated with either 1% or 1.5% chitosan (CS) or chitosan combined with calcium gluconate (CaGlu). Following treatment, strawberries were stored at 10 °C and 70 ± 5% RH for one week. The effectiveness of the treatments in extending fruit shelf-life was evaluated by determining fungal decay, respiration rate, quality attributes and overall visual appearance. No sign of fungal decay was observed during the storage period for fruit coated with 1.5% CS (with or without the addition of CaGlu) or 1% CS + 0.5% CaGlu. By contrast, 12.5% of the strawberries coated with 1% CS lacking calcium salt were infected after five days of storage. The chitosan coating reduced respiration activity, thus delaying ripening and the progress of fruit decay due to senescence. Chitosan coatings delayed changes in weight loss, firmness and external colour compared to untreated samples. Strawberries coated with 1.5% chitosan exhibited less weight loss and reduced darkening than did those treated with 1% chitosan, independently of the presence or absence of CaGlu. However, addition of calcium to the 1% chitosan solution increased the firmness of the fruit. Coated samples had greater visual acceptability than had untreated fruits. The addition of calcium gluconate to the chitosan coating formulation increased the nutritional value by incrementing the calcium content of the fruit.     

Antifungal activity of sodium acetate and Lactobacillus rhamnosus

The inhibition of molds by sodium acetate in deMan Rogosa Sharpe (MRS) medium, along with the antifungal activity of Lactobacillus rhamnosus VT1, was studied by the slope agar plate method. MRS agar prepared with and without sodium acetate was used as the agar substrate. A total of 42 strains of Aspergillus, Penicillium, Fusarium, Alternaria, Cladosporium, and Rhizopus were used to compare sensitivities to the inhibitory activity of sodium acetate and L. rhamnosus VT1. It was found that sodium acetate in MRS medium affected the growth of 33 of the 42 mold strains tested to various degrees. The highest sensitivity to sodium acetate was shown by strains of Fusarium, followed by strains of Penicillium, Aspergillus, and Rhizopus. L. rhamnosus VT1 also inhibited mold growth. A significant finding was that sodium acetate and L. rhamnosus VT1 in combination exhibited a possible synergistic action. Thirty-nine of the 42 mold strains tested were completely inhibited by the presence of both antifungal agents. This finding confirms that sodium acetate, a basic component of commercial MRS medium, has strong antifungal properties, and this must be taken into consideration when evaluating the antifungal activity of Lactobacillus cultures grown in MRS broth.     

Effect of antifungal hydroxypropyl methylcellulose-lipid edible composite coatings on Penicillium decay development and postharvest quality of cold-stored "Ortanique" mandarins

Edible composite coatings based on hydroxypropyl methylcellulose (HPMC), hydrophobic components (beeswax and shellac), and food preservatives with antifungal properties were evaluated on "Ortanique" mandarins during long-term cold storage. Selected food preservatives included potassium sorbate (PS), sodium benzoate (SB), sodium propionate (SP), and their mixtures. Intact mandarins or mandarins artificially inoculated with the pathogens Penicillium digitatum and Penicillium italicum, the causal agents of citrus postharvest green (GM) and blue (BM) molds, respectively, were coated and stored up to 8 wk at 5 °C + 1 wk of shelf-life at 20 °C. HPMC-lipid coatings containing food preservatives controlled better GM than BM on Ortanique mandarins. SB- and SB + SP-based coatings reduced the incidence of GM by about 35% after 4 wk at 5 °C. Among all coatings, only the SB-based coating reduced the incidence of GM (about 16%) after 6 wk at 5 °C. All coatings significantly reduced disease severity of both GM and BM after 6 wk at 5 °C. Analytical and sensory fruit quality was evaluated on intact mandarins. All coatings, especially the SB + SP-based coatings, were effective to control weight loss and maintain the firmness of coated mandarins. Internal gas concentration, juice ethanol and acetaldehyde content, sensory flavor, off-flavor, and fruit appearance were not adversely affected by the application of the antifungal coatings. Further studies should focus on the modification of some physical characteristics of the coatings to improve the gloss and visual aspect of treated mandarins.     

Effect of shrimp chitosan coatings as affected by chitosan extraction processes on postharvest quality of strawberry

Edible coatings as affected by chitosan extraction processes were used to preserve the quality of strawberries (Fragaria × ananssa) during their storage at ambient temperature (20–25 °C). Thus, three different chitosans were prepared from shrimp shell and were designated as the following: C1 by classical method, C2 without the decoloration step, and C3 without the decoloration step and the deproteinization step. In order to study the effectiveness of coatings, changes in physicochemical parameters and mold spoilage were studied. Chitosan coatings had no significant effects on titrable acidity, pH and soluble solids content (SSC) of strawberries throughout the storage, while the SSC content of control fruits increased with the storage time. In contrast, chitosan coatings delayed changes in weight loss and the appearance of fungal infection. Coated samples had greater visual acceptability than had the untreated fruits. By visual analysis, it was possible to verify that the best quality was maintained until the day 12, for the strawberries coated with C3 (1 %).     

Effects of chitosan-oil coating on blue mold disease and quality attributes of jujube fruits

The effects of chitosan coating enriched with cinnamon oil on blue mold disease and quality attributes were investigated. In the in vitro experiment, the results demonstrated that the antifungal activity against P. citrinum improved with increasing concentration of chitosan or cinnamon oil. In the in vivo study, chitosan-oil treatments significantly reduced fungal decay caused by P. citrinum and all compounds with cinnamon oil at 2.0% showed complete control of the growth of P. citrinum on wound-inoculated fruits. High chitosan-oil concentrations correlated with low disease incidence regardless of storage temperature. Treatments of chitosan-oil coating also inhibited the activity of polyphenol oxidase and maintained vitamin C and phenolic compounds in wounded jujube fruits. Results suggested that the effect of chitosan coating (1.0%) enriched with cinnamon oil (0.75%) on blue mold in jujube fruits may be associated with fungitoxic properties against the pathogen and the elicitation of biochemical defense responses in fruits.     

Antimicrobial Action of Chitosan against Spoilage Organisms in Precooked Pizza

ABSTRACT: Chitosan antifungal capacity when the biopolymer is used as an edible film and as a dough ingredient in precooked pizza was studied. This action was evaluated against the fungi that produce major spoilage in that product. It was observed that the use of chitosan in acetic acid as edible coating (0.079 g/100 g pizza) delayed Alternaria sp, Penicillium sp, and Cladosporium sp (Deuteromycetes) growth. This behavior was similar compared with the action of calcium propionate (0.103 g/100 g pizza) and potassium sorbate (0.034 g/100 g pizza) preservatives. On the other hand, chitosan showed a little sensibility on Aspergillus sp (Zygomycetes). It was demonstrated that the use of chitosan into the dough was not effective.     

Antifungal activity of edible coating made from chitosan and lactoperoxidase system against Phomopsis sp. RP257 and Pestalotiopsis sp. isolated from mango

Lactoperoxidase system (LPOS) was incorporated into chitosan solutions (0.5, 1, and 1.5%) to protect mangoes against two strains of fungi. Coating solutions effectiveness in vitro (in Petri dish) and in vivo (on mango) were studied on fungal (Phomopsis sp. RP257 and Pestalotiopsis sp.) growth isolated from mango cv Amelie. In vitro, chitosan concentration at least 1% containing or not LPOS effectively inhibited Pestalotiopsis sp. growth at 100%. Presence of LPOS or Lactoperoxydase system with iodine (LPOSI) in chitosan at 0.5% increased the percentage of inhibition from 26 to 93%. Edible films with LPOS inhibited Phomopsis sp. RP257 particularly when LPOS was incorporated in chitosan concentrations of 1 and 1.5%. Iodine did not influenced antifungal activity of LPOS against Pestalotiopsis sp. but decreased activity antifungal toward Phomopsis sp. RP257. The properties (water vapor permeability and mechanical properties) of chitosan films were not significantly changed by the incorporation of the enzyme system. in vivo condition, chitosan coating at 1 and 1.5% with or without enzyme system was sufficient to inhibit totally (100%) Pestalotiopsis sp. and was 60% efficient against Phomopsis sp. with chitosan only at 1 and 1.5%. However, when coating solution mainly at 1 and 1.5% was enhanced by LPOS with or without iodine, it inhibited totally (100%) Phomopsis sp. RP257. The presence of iodine slightly reduced antifungal activity against Phomopsis sp. RP257.     

Effects of Antimicrobial Edible Coating of Thymol Nanoemulsion/Quinoa Protein/Chitosan on the Safety,Sensorial Properties,and Quality of Refrigerated Strawberries (<Emphasis Type="Italic">Fragaria × ananassa</Emphasis>) Under Commercial Storage Environment

The effectiveness of the edible coating with thymol nanoemulsion on the safety, sensorial properties, and quality of refrigerated strawberries was investigated under commercial storage conditions. Spontaneous emulsification was used to obtain the thymol nanoemulsion that was included on quinoa protein/chitosan coatings. During the entire storage time, strawberries coated with thymol-antimicrobial packaging had a lower fungal and yeast load compared with the controls (uncoated and coated with quinoa protein/chitosan). The flavour and aroma of the coated strawberries was initially affected, although this sensory appreciation was improved from the fifth day of storage and showed similar scores than the controls, and presenting better aroma score at day 12 of storage. Furthermore, the shelf life of the thymol nanoemulsion-coated strawberries increased in 4 days, unlike that in the both controls. Further, the application of these biocoatings on strawberries significantly decreased the weight loss relative to that in the control, during 16 days of storage at 5 °C and 90% relative humidity, and did not alter the quality parameters (pH, titrable acidity, and percentage of soluble solids). These results suggest that the application of thymol/nanomulsion-loaded edible films is an effective strategy to increase the shelf life of highly perishable products such as strawberries.     

Edible Active Coatings Based on Pectin,Pullulan, and Chitosan Increase Quality and Shelf Life of Strawberries (Fragaria ananassa)

Edible active coatings (EACs) based on pectin, pullulan, and chitosan incorporated with sodium benzoate and potassium sorbate were employed to improve the quality and shelf life of strawberries. Fruits were washed, disinfected, coated by dipping, packed, and stored at 4 °C for 15 d. Application of EACs reduced (P < 0.05) weight loss and fruit softening and delayed alteration of color (redness) and total soluble solids content. In contrast, pH and titratable acidity were not affected (P > 0.05) throughout storage, and ascorbic acid content was maintained in pectin‐EAC coated strawberries. Microbiological analyses showed that application of EACs reduced (P < 0.05) microbial growth (total aerobic counts, molds, and yeasts) on strawberries. Chitosan‐EAC coated strawberries presented the best results in microbial growth assays. Sensory quality (color, flavor, texture, and acceptance) improved and decay rate decreased (P < 0.05) in pectin‐EAC, pullulan‐EAC, and chitosan‐EAC coated strawberries. In conclusion, EACs based on polysaccharides improved the physicochemical, microbiological, and sensory characteristics, increasing the shelf life of strawberries from 6 (control) to 15 d (coated fruits).     

Mycoflora of two types of Portuguese dry-smoked sausages and inhibitory effect of sodium benzoate, potassium sorbate, and methyl p-hydroxybenzoate on mold growth rate

The mycoflora of chouriqo types Alentejano and Ribatejano, two varieties of Portuguese dry-smoked sausages, have been investigated after a producer-defined shelf life period (120 days at 20 +/- 5 degrees C) in modified atmosphere packaging (55% N2 and 45% CO2). On the basis of morphological and physiological characteristics, the isolates were identified as Penicillium, Aspergillus, Fusarium, Rhizopus, Monilia, Absidia, and Cephalosporium. The species identified were as follows: Penicillium terrestres (43.4%), Penicillium sp. (13.3%), Fusarium sp. (10%), Aspergillus glaucus (10%), Aspergillus versicolor (6.8%), Monilia fruticola (3.3%), Absidia sp. (3.3%), Cephalosporium sp. (3.3%), Rhizopus stolonifer (3.3%), and Fusarium tricinctum (3.3%). Additionally, the effects of three preservatives (potassium sorbate [PS], sodium benzoate [SB], and methyl p-hydroxybenzoate [MHB]) were studied on the growth rate of mold representative isolates. MHB showed a greater inhibitory effect than SB and PS in all fungi isolates, except in A. glaucus [Tm30(A)], in which the inhibitory effect of MHB was similar to PS. At 0.05% (wt/vol), all fungi were inhibited with MHB, except for R. stolonifer [Tm25(A)], which started to decrease the growth rate only at a concentration higher than 0.1%. PS was more effective at inhibiting mold growth than SB, except in Absidia sp. [Tm16(R)], in which both showed a similar inhibitory effect. MHB showed great promise as an application to the surface at 0.1% (wt/vol) to improve the stability and safety of the product through the inhibition of potential spoilage and toxigenic molds.     

Effect of Edible Coating Process and Irradiation Treatment of Strawberry fragaria spp. on Storage-keeping Quality

ABSTRACT: Gamma-irradiation and various edible coatings were tested on fresh strawberries ( Fragaria spp. ) for keeping fruit quality and extending shelf life. Four coatings based on milk protein were evaluated. In 1 experiment, coating formulation based on caseinate and/or strawberries were irradiated using a ⁶⁰Co source. Both gamma-irradiation treatment and edible coating process significantly delayed (p ≤ 0.05) molds growth. Edible coating based on irradiated caseinate was more effective than that of unirradiated caseinate. In a 2nd experiment, 3 irradiated coatings based on calcium caseinate and whey proteins were evaluated. The coating formulation based on 1:1 caseinate-whey was found to be more effective than those based on calcium caseinate. Addition of calcium chloride or a mixture of pectin and agar increased the effectiveness of the coating by delaying molds' apparition.     

Performance of hydroxypropyl methylcellulose (HPMC)-lipid edible coatings with antifungal food additives during cold storage of ‘Clemenules’ mandarins

The performance of edible composite coatings containing hydroxypropyl methylcellulose (HPMC), hydrophobic components (beeswax and shellac), and food preservatives as antifungal ingredients was evaluated on ‘Clemenules’ clementine mandarins. Tested preservatives included potassium sorbate (PS), sodium benzoate (SB), sodium propionate (SP), and their mixtures. Intact fruit or fruit artificially inoculated with Penicillium digitatum or Penicillium italicum, the causal agents of citrus postharvest green (GM) and blue (BM) molds, respectively, were coated and stored up to 30 d at 5 °C plus 7 d at 20 °C of shelf-life. During cold storage, all HPMC-lipid coatings containing food additives significantly reduced the development of both GM and BM, although the performance was better against GM. When coated fruit were transferred to 20 °C, all coatings lost effectiveness. SB + PS-based coating was the most effective to reduce disease severity. All the coatings effectively reduced weight loss and maintained rind firmness of coated ‘Clemenules’ mandarins. The coatings did not adversely affect the ethanol content of the juice, sensory flavor, and fruit appearance. Although the internal gas concentration of coated fruit was modified, the coatings did not induce off-flavor.     

纳米SiO2壳聚糖复合膜保鲜草莓的研究

优化壳聚糖纳米SiO2复合膜透CO2性,并将其应用于草莓保鲜实验.结果表明,壳聚糖复合膜最佳配方为:壳聚糖含量2 g,纳米TiO2含量0.07g,冰乙酸含量1.4mL,且透CO2量达到最低为0.0909g/d;优化膜处理的草莓常温下贮藏6d后,腐烂指数比空白组降低了5.1%;低温4℃下贮藏11 d后,腐烂指数比空白组降...