Considering the widespread use of polymer composites due to their diverse engineering properties, the incorporation of natural fibers is a novel approach to enhance their environmental properties and acquire additional engineering benefits. Applying natural fibers in composites can offer a biocompatible solution to plastic and polymer composite waste issues. Studying the overall properties of composites made with these fibers is a novel area of research. Investigating effective methods for detecting defects and measuring mechanical properties, such as integrity, resilience to failure, and fracture toughness, is particularly crucial. In the current study, the effect of adding cotton and kenaf natural fibers on the resistance to delamination, which is the most common type of failure in laminated composites, is investigated in mode Ⅱ of loading. Specimens of woven glass/epoxy composites with natural fibers were fabricated using the hand lay-up method and end notched three-point bending test was performed. Using the methods provided in the JIS K 7086 standard, the moment of initiation of delamination is determined in the samples. The results demonstrate that only using the mechanical methods provided in the mentioned standard is not sufficient to determine the initiation of delamination. Hence, the acoustic emission can be employed for an easier and more accurate determination of the initiation of delamination. According to the obtained results, cotton and kenaf fibers show a significant increase in the interlaminar fracture toughness of woven glass/epoxy composites. Using cotton fibers shows 212% − 249% and employing kenaf fibers shows 144% − 281% increase in interlaminar fracture toughness respectively. Among the different acoustic emission data analytics, the cumulative energy method shows the lowest mean percentage deviation error of %2.59 among other examined diagnosing techniques.