| Summary: | Recently, there have been urgent economic and scientific demands to decrease the use of chemical fungicides during the treatment of phytopathogens, due to their human health and environmental impacts. This study explored the biocontrol efficacy of novel and eco-friendly preen (uropygial) oil and endophytic <i>Bacillus safensis</i> in managing postharvest <i>Botrytis</i> grey mold in strawberry fruit. The preen oil (25 μL/mL) showed high antifungal activity against <i>B</i>. <i>cinerea</i> Str5 in terms of the reduction in the fungal radial growth (41.3%) and the fungal colony-forming units (28.6%) compared to the control. A new strain of <i>Bacillus safensis</i> B3 had a good potential to produce chitinase enzymes (3.69 ± 0.31 U/mL), hydrolytic lipase (10.65 ± 0.51 U/mL), and protease enzymes (13.28 ± 0.65 U/mL), which are responsible for the hydrolysis of the <i>B</i>. <i>cinerea</i> Str5 cell wall and, consequently, restrict fungal growth. The in vivo experiment on strawberry fruit showed that preen (uropygial) oil reduced the disease severity by 87.25%, while the endophytic bacteria <i>B</i>. <i>safensis</i> B3 reduced it by 86.52%. This study reports the efficiency of individually applied bioagents in the control of phytopathogenic fungi for the first time and, consequently, encourages their application as a new and innovative strategy for prospective agricultural technology and food safety.
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