Probiotics are considered as desirable alternatives to antibiotics because of their beneficial effects on the safety and economy of farm animals. The protectant can ensure the viability of probiotics, which is the prerequisite of the beneficial effects. The objective of this study was to evaluate the effects of a novel nano-protectant containing trehalose, skim milk powder, phytoglycogen nanoparticles, and nano-phytoglycogen Pickering emulsions on the viability of K17 under different conditions. The results indicated that the optimal concentration of the carbohydrate substrate was determined to be 10% skim milk powder (/) instead of trehalose. The combination of 10% skim milk powder (/), 1% phytoglycogen nanoparticles (/), and 10% Pickering emulsions (/) was selected as the optimal component of the protectant. Trilayer protectants with an optimal component had a more significant protective effect on the bacteria than that of the monolayer and bilayer protectants, or the control in feed storage, freeze-drying, and simulated gastrointestinal environment. A scanning electron microscope was used to monitor the morphological characteristics of the protectants for different layers on . In conclusion, the trilayer protectant exhibited a substantial effect on during storage and consumption, which could be used in the feed and functional food.