The Efficiency of Cold Plasma Treatment on Aspergillus Niger Disinfection and Quality Properties Preservation of Grapes during the Post-harvest Period

ماشین های کشاورزی

Volume 14, Issue 3 - Serial Number 33, September 2024, Pages 337-353   

Document Type: Research Article

DOI: 10.22067/jam.2023.84550.1191

Authors

A. Khalaj¹; E. Ahmadi^{* 1}; S. Mirzaei²; F. Ghaemizadeh³; R. Abbaszadeh⁴

¹Department of Biosystems Engineering, Faculty of Agriculture, Bu-Ali Sina University, Hamadan, Iran

²Department of Plant Protection, Faculty of Agriculture, Bu-Ali Sina University, Hamadan, Iran

³Department of Horticultural Science, Faculty of Agriculture, Bu-Ali Sina University, Hamadan, Iran

⁴Iranian Research Organization for Science and Technology, Tehran, Iran

Abstract

Introduction
Grape is a major horticultural crop with a high nutritional value in the world. The optimal geographic and climatic conditions in Iran have positioned it as one of the most important regions for grape cultivation in the world. Black rot, caused by Aspergillus niger, is a highly destructive fungal disease that affects the grapes by targeting wounded areas. It causes crushing of the grapes, the falling of berries, and reduces transportation and storage properties (Ponsone et al., 2011). Various methods, such as fumigating bunches with sulfur dioxide and storing them in a modified atmosphere, have been used to control fungal rot and prolong the shelf life of grapes. However, each method has its limitations. Grape fumigation with sulfur gas is harmful to human health (Duarte-Sierra et al., 2016) and the efficiency of modified atmospheric storage on rot control and maintaining the quality of grapes depends on the type of variety, storage temperature, and especially gas concentration (Himelrick, 2003).
Given the lack of efficiency in traditional methods, it is imperative to introduce modern techniques that can effectively disinfect microorganisms. These advanced methods offer several advantages, including the preservation of crop quality, an increase in crop shelf life, the promotion of good health, and substantial economic benefits. A technique of this type includes using non-thermal (cold) plasma (NTP) technology to eliminate food microorganisms (Bourke et al., 2018). The effect of cold plasma at atmospheric pressure on the reduction of bacterial populations in food products such as lettuce, tomato, strawberry, and cherry tomato has been reported (Bermúdez-Aguirre et al., 2013; Pasquali et al., 2016; Ziuzina et al., 2014). Research has shown that cold plasma can effectively inactivate Aspergillus in various orchard and agricultural products (Butscher et al., 2016; Ghorashi et al., 2020; Selcuk et al., 2008). The effect of cold plasma on the quality characteristics of the product during the post-harvest period has also been investigated. Blueberries treated with cold plasma for less than 15 minutes showed remarkable results: after 10 days, the fruit exhibited reduced lipid peroxidation and darkening, with no impact on the total anthocyanin content, pH, or firmness of the product (Hu et al., 2021). In a study by Rana et al. (2020), it was found that subjecting strawberries to 15 minutes of cold plasma with packaging after 5 days of storage at 25°C had no significant impact on pH, TSS, and moisture content of the fruit.
The review of the literature reveals the absence of research on fungal disease control and grape quality evaluation following the use of NTP. This study aims to investigate the efficiency of plasma treatment in reducing the infection with Aspergillus fungi, along with examining the physical, chemical, and mechanical properties of Fakhri grape.
Materials and Methods
This research was conducted as a completely randomized design in a factorial experiment at four plasma levels (0, 10, 20, and 40 s) and five storage periods (1, 2, 3, 4, and 5 weeks) with three replications at 4°C.  A plasma generator was first designed and manufactured in this study. A specifically designed and fabricated plasma application probe was also developed for grape berries. The individual grape berries were then sterilized with 1% sodium hypochlorite under a laminar hood for 2 minutes. Afterward, they were rinsed three times with sterile distilled water to remove any remaining disinfectant residue from their surfaces. Sterilized berries were immersed in Aspergillus spore suspension with 105 spores.ml^-1 concentration. Finally, all samples were dried on paper filters and prepared for different plasma treatment durations (0, 10, 20, and 40 s). The treated samples were stored at 4°C, and the infection percentage and microbial load were measured on a weekly basis. To assess the preservation quality, chemical parameters such as pH, TSS, and TA, physical parameters (color change and weight loss), and mechanical properties were measured every week. Additionally, thermal imaging was performed weekly.
Results and Discussion
Plasma application during storage significantly reduced the infection percentage and microbial load in Aspergillus-inoculated samples. At the end of the storage period, the infection percentage and microbial load in the 40-second plasma treatment were 5% and 2.5 CFU g^-1 respectively, while in the control group, the infection percentage was 100% and the microbial load was 4 CFU g^-1. At the end of the storage period, the lowest pH level in the plasma was observed for 40 s plasma. This could be attributed to effective contamination control, as fungal contamination leads to alkalization of the environment. The highest amount of TSS was also observed in control and 40 s plasma. But in the 10 and 20 s plasma treatment, the process of changes was gradual and not significant. The higher TSS level of control and 40 s plasma can be due to the weight loss caused by the spread of contamination and moisture leakage caused by damage to the tissue. This decrease in moisture leads to an apparent increase in the TSS index. Research has shown that plasma primarily affects the surface characteristics of products, and when applied with the appropriate voltage and duration, it does not alter the internal chemical properties (Hu et al., 2021). Over time, weight loss increased in all treatments. This increasing trend during the storage period is higher in control and 40 s plasma compared to 10 and 20 s plasma. Therefore, the weight loss in the control can be due to the spread of contamination and aging of the product over time. However, the weight loss in the 40-second plasma treatment can be due to the destruction of the fruit tissue caused by longer duration of the plasma application.
In the current research, by increasing the duration of plasma application to 40 s, a significant decrease in L*, a*, and b* indices and an overall change in the color of the product was observed. Research shows that in blueberries, inappropriate duration of plasma treatment causes the loss of wax on the fruit surface and leads to darkening of the product (Hu et al., 2021). The highest and lowest changes in temperature drop were observed in the control treatment (5°C) and 10 and 20 s plasma (3 and 3.5°C, respectively). According to research, an increase in fungal contamination leads to a decrease in humidity, increases weight loss, and subsequently a decrease in product temperature. A decline in mechanical characteristics was noted for the control and plasma treated samples during the storage period. The lowest value for indicators was observed in the 40 s plasma treatment. However, no significant difference was observed in samples treated with plasma for less than 20 seconds compared to the control group up to the middle of the storage period. According to a report by Misra et al. (2014), plasma application can reduce tissue softness. Therefore, optimizing its plasma duration and intensity is very important (Pan et al., 2021).
 Conclusion
Our experiments aimed to investigate the effect of NTP treatment on controlling Aspergillus infections while preserving the quality properties of Fakhri grapes. The obtained results are important for two main reasons. Firstly, an innovative probe was designed for plasma applications, specifically tailored to the shape and size of individual grapes in order to thoroughly cover them with plasma. Secondly, application of plasma was carried out for the first time and yielded valuable results, indicating that this technique can control fungal infections and preserve the chemical, physical, and mechanical properties of grapes.

Keywords

Chemical properties; Cold plasma; Mechanical properties; Microbial load; Physical properties

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