Antifungal Activity of Squid Pen Chitosan Nanoparticles against Three Fungal Pathogens in Various Citrus Fruits In Vitro and In Vivo

Fungal infections have been considered a primary cause of the postharvest losses of citrus fruits. Therefore, it is necessary to find low-cost and high antifungal activity materials for preventing the decay of citrus fruits after harvest. In this study, squid chitosan nanoparticles (SCNs) were prepared from squid pen chitosan and used as a biofungicide against three citrus fungal pathogens in both in vitro and in vivo evaluations. The prepared SCNs had a mean size of ca. 56 nm and a high zeta potential of +98.7 eV with a narrow size distribution. At a range of 50–250 ppm, the SCN concentration of 200 ppm exhibited the highest activity in totally inhibiting the growth of Lasiodiplodia pseudotheobromae, Alternaria alternate, and Penicillium digitatum in in vitro tests where these fungi were isolated from symptomatic fruits and identified. Furthermore, after 12 days of incubation at 30 ± 0.2 °C and high relative humidity in in vivo studies, the infection area of the sample treated at 250 ppm completely suppressed fruit disease symptoms. The results demonstrate that prepared SCNs efficiently control postharvest citrus fruit diseases. These findings recommend applying SCNs as a potential candidate for citrus fruit storage.