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Background  Plants of  Allium  spp., including garlic ( A. sativum ) and onions  (A. cepa ), are known to be oxidatively toxic to canine erythrocytes resulting in Heinz body hemolytic anemia in dogs. In humans, these plants have been used as medicinal agents for multiple diseases since ancient times. Especially, fresh garlic extracted over a prolonged period produces less irritative and odorless aged garlic extract (AGE), containing unique and beneficial organosulfur compounds that can help prevent many kinds of diseases. In this study, the safety and efficacy of long-term oral administration of AGE is evaluated in dogs. The objectives are to confirm the safe dosage for long-term use and beneficial functions of AGE for dogs and to plan and design a canine health supplement or a preventive agent for multiple diseases based on the data of this study.    Results  Beagles were orally administered AGE (45 or 90 mg/kg body weight once a day) or an equivalent amount of water as control for 12 weeks. In AGE-treated groups, at 12 weeks post-administration at a dose of 90 mg/kg, there were no observable changes in the clinical signs, complete blood count, and serum biochemical parameters. Heinz bodies and eccentrocytes, the markers of oxidative damage in erythrocytes, did not appear in blood smear examination. In order to further evaluate the beneficial effects of AGE on health of dogs, the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) gene ( NFE2L2 ) and Nrf2-regulated phase II antioxidant enzyme genes ( NQO1 ,  GCLM ,  HMOX1 , and  SOD2 ) were determined in whole blood between pre- and post-AGE administration. The expression of  NFE2L2  gene was significantly upregulated in the AGE-treated groups [45 ( p  < 0.05) and 90 mg/kg ( p  < 0.01), 8 weeks] as compared to in the control group. Among the Nrf2-regulated enzymes examined, the expressions of  NQO1  [45 ( p  < 0.05) and 90 mg/kg ( p  < 0.01), 8 weeks] and  GCLM  [45 ( p  < 0.05) and 90 mg/kg ( p  < 0.01), 12 weeks] genes were significantly upregulated.    Conclusion  The long-term oral administration of AGE at a dose of 90 mg/kg/day for 12 weeks did not show any adverse effects in dogs. Furthermore, the administration of AGE upregulated the gene expressions of canine Nrf2 and Nrf2-regulated phase II antioxidant enzymes. These results suggest that AGE might safely contribute to the health of dogs provided that the appropriate dosage is used.    Electronic supplementary material  The online version of this article (10.1186/s12917-018-1699-2) contains supplementary material, which is available to authorized users.

bmc veterinary research

Safety and efficacy of aged garlic extract in dogs: upregulation of the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway and Nrf2-regulated phase II antioxidant enzymes