Oil spills can result in significant damage to marine estuaries, rivers, lakes, wetlands, and shorelines. Electrospun nanofibers containing biosurfactant (ENFs) can be used to clean oil spills up and protect the environmental biology. Present work aimed to study the side-effects of prepared nanofibers on animal models. Screening of the prepared ECNFs on animals showed that three of them (PVA-5, PEO-1, and PEO-5) are safe to hepatic tissues and liver functions. Furthermore, oxidative stress did not change after using these nanofibers. The PVA-1 nanofibers, however, were found to cause major pathological changes in the liver tissue. In addition, PVA-1 nanofibers were proved to alter the total white blood count and the neutrophil percentages significantly in comparison to the control. In conclusion, PVA-5, PEO-1, and PEO-5 are safe to hepatic tissues and liver functions.