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Identifying sustainable resources of natural fibers is essential due to their high demand in industrial applications such as automotive and biomedical materials. Two alternative fibers obtained from canola and sweet clover stalks were characterized for their properties using energy dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), contact angle, and tensile test. Hemp and flax fibers, both in use as industrial fibers, were also characterized as conventional fibers. Results showed that all the fibers had the same chemical elements (carbon, oxygen, magnesium, and potassium) and chemical bonds. The crystallinity index for the alternative fibers ranged from 62 to 71%, which was close but lower than the conventional fibers (82% for hemp and 80% for flax). The thermal stability of the alternative fibers was around 220 °C, close to the conventional fibers (230 °C). The alternative fibers had contact angles of less than 90°, showing high surface energy. Since the alternative fibers had a low Young's modulus and tensile strength (5.57–8.52 GPa and 57.45–71.26 MPa, respectively), they are suitable for some specific applications in the biomedical industry. In contrast, conventional fibers are suitable where a higher stiffness and strength is required. [ABSTRACT FROM AUTHOR]