<b>Background and Objective:</b> Typically, free-living amoebae, members of the genus <i>Acanthamoeba</i> can exist in a range of biological niches, such as fresh and brackish water, filters for heating, soil, airborne dust, ventilation, in addition to air conditioning, as well as pools and hot tubs. On rare occasions, these can be linked to infections of the central nervous system in both people in addition to animals. This study aimed to identify the cause of rats' death by isolating and identifying <i>Acanthamoeba</i> sp., from the ovary of Sprague-Dawley rats. <b>Materials and Methods:</b> An amoeba was identified and cultured from the ovary of Sprague-Dawley rats that succumbed to an amoebic illness. Considering culture attributes, growth shape and immunofluorescence measures, <i>Acanthamoeba</i> sp., was determined as the infecting amoeba. <b>Results:</b> <i>Acanthamoeba</i> sp., the contaminating single adaptable cell, was discovered. Furthermore, by sequencing a symptomatic part of the atomic little subunit ribosomal RNA gene to recognize the one-celled critter as <i>Acanthamoeba</i> sp., <b>Conclusion:</b> An <i>Acanthamoeba</i> strain was demonstrated that has a wide range of genotypes and is capable of infecting people and other species can also infect rats fatally.
Keywords: Acanthamoeba; Sprague-Dawley rats; isolation; molecular identification; morphology; ovary.