Chitosan is one of the most abundant biopolymers in nature with high economic value due to its biocompatibility, biodegradability, lack of toxicity, and antifungal activity. In this study, chitosan was extracted from three different sources: Blaps lethifera (CSB), Pimelia fernandezlopezi (CSP), and Musca domestica (CSM). The ash content (AC), moisture content (MC), fat binding capacity (FBC), water binding capacity (WBC), and deacetylation degree (DD) were determined for the prepared chitosans. The effect of the DD of chitosan on the antibacterial activity of gram (positive/negative) bacteria and the azo dyes (methylene blue, MB) removal from wastewater was also investigated. Chitosan extracts showed good antibacterial activity against Listeria innocua, Bacillus subtiliis, Staphylococcus aureus, Salmonella typhimurium, and Pseudomonas aeruginosa. The MB dye removal of CSB-chitosan, CSP-chitosan, and CSM-chitosan reached 37%, 87%, and 26%, respectively, at a contact time of 2 h, a low initial dye concentration MB of 13 ppm, a solution temperature of 25 °C, and a pH = 7.Graphical