In this study, Difethialone 0.0025 % in diet within 3 days non-choice feeding test caused complete mortality for R. norvegicus and R. rattus indicating that they were susceptible to Difethialone. The bait consumption and corresponding active ingredient intake was more in R. rattus than R. norvegicus. The mean intake values for R. rattus being 3.1 and 2.64 mg/kg group A and 2.6 and 2.65 mg/kg group B for males and females, respectively and for R. norvegicus 1.93 and 2.29 mg/kg at group A and 2.14 and 2.09 mg/kg group B for males and females, respectively. Within one day non-choice feeding test the obtained results showed that it caused mortality ranged between 60–70 % for R. norvegicus and 50–60 % for R. rattus for both sexes.The higher intake values of active ingredient for died rats recorded to the males of R. rattus group A (1.45 mg/kg) while males of R. norvegicus group A recorded the lower value (0.77 mg/kg).The higher mean death time was recorded to the males of R. norvegicus group B (8.5 days), the lower value was recorded for females of R. rattus group B that the more of feeding days test, Difethialone within 2 days non-choice feeding test caused mortality between 70-80 % for both species at two regions. The higher values were recorded for male of R. rattus group A (2.25 mg/kg), while the lower values were recorded for female of R. norvegicus group B (1.39 mg/kg). The mean values of time to death showed that male of R. rattus recorded the higher values (6.16 days), while females of R. norvegicus showed the lower value group A (4.75 days).