The present study aimed to evaluate the effect of Aloe vera addition on the viability and antifungal activity of TEP6 (Lactobacillus paracasei) bacteria immobilized on chitosan films for 28 days. Different chitosan and A. vera proportions and carbon sources at several pH values were tested as formulations for supporting the microorganism. Bacterial viability was maintained in freshly made films, with values of 10.4, 10.8 and 10.9 log CFU·g⁻¹ for the formulations containing 70% (T11), 100% (T8) and 100% (T16) of A. vera, respectively. The same formulations (T8, T11 and T16) maintained bacterial viability for 14 days of film storage with a loss to values of 9.5 log CFU·g⁻¹. By applying a quarter fraction 2^5–2 experimental design with an array of five factors, the factors with the greatest influence on viability and antifungal activity were determined. The optimal conditions for viability were the formulation with 100% A. vera, pH 4.5 and 0.1 M glucose. The antifungal activity of fresh films was influenced by the formulation with 10 g·L⁻¹ glycerol and 100% A. vera, showing a 60% inhibition of fungal (Colletotrichum gloeosporioides) growth. The films developed in this study may have the potential to be used as coatings on vegetal products susceptible to attack by Colletotrichum gloesporioides.