Entomophagy, the practice of consuming insects, is a global tradition. In Mexico, one of the most notable and widely consumed insects is the larva of Aegiale hesperiaris. This insect feeds on the leaves of various Agave species with high polysaccharide content, suggesting their potential role as prebiotics for the intestinal microbiota, particularly lactic acid bacteria (LAB). LAB are recognized for their use as probiotics in foods due to their health-promoting capabilities. In this study, LAB from the intestinal microbiota of A. hesperiaris larvae were isolated and characterized, utilizing 16S rRNA gene identification. The analysis revealed three bacterial species from the Lactobacillaceae family, indicating a close symbiotic relationship with the insect. This suggests a significant impact on carbohydrate and protein metabolism, vitamin synthesis, and amino acid production, contributing to the high nutritional value of this edible insect. The study provides insights into the bacteria within the digestive tract of A. hesperiaris larvae and their role in enhancing the nutritional value of this edible insect. Additionally, it establishes a foundation for future research on the ecological roles and potential biotechnological benefits of these bacteria in the food industry and the development of therapies for various conditions and diseases.