Aflatoxins (AFs) are a group of potent mycotoxins that frequently pollute agro-products and animal feeds. For complex reasons, the regulatory limits of AFs differ from nation to nation. In this study, feedstuffs contaminated with various concentrations of aflatoxin B1 (AFB1) corresponding to AF regulatory limits of major agro-product importing and exporting nations were tested in Sprague-Dawley rats, and the toxicological results were evaluated. During the trial period, no poisoning behavior was observed and no animals died. No significant differences in body weight were observed between AFB1-contaminated and control rats. However, various negative impacts on liver and kidney functions were observed in AFB1-contaminated groups, and these negative impacts were positively correlated with AFB1 concentrations. At concentrations of 4-50 µg kg(-1) , AFB1 tolerance was observed for the liver after 15 days of testing, but this tolerance was not observed for the kidney. Low concentrations of AFB1 still induced obvious toxicological impacts on Sprague-Dawley rats, and the negative effects were aggravated by increasing the AFB1 dose and feeding time. Therefore strict AF regulatory limits are suggested for food safety administration, especially for the low-income nations with higher hepatitis B virus prevalence.