Acute Oral Toxicity Test of a Combination of Citronella (Cymbopogon nardus) and Neem (Azadirachta indica) Oils in Female DDY Mice

Background: Increasing interest in plant-based veterinary products requires scientific evaluation of their safety. Citronella (Cymbopogon nardus) and neem (Azadirachta indica) oils contain bioactive compounds with antimicrobial and antiparasitic activities; however, data regarding the acute toxicity of their combination remain limited. Aims: This study aimed to evaluate the acute oral toxicity and safety profile of a 1:1 combination of citronella oil (Cymbopogon nardus) and neem oil (Azadirachta indica) in female DDY mice. Methods: A total of 24 female DDY mice were divided into four groups: one control group (aquabidest) and three treatment groups, each receiving a 1:1 mixture of citronella and neem oils at doses of 5, 10, and 15 g/kg body weight (BW) via a single oral administration. The observations were carried out for 14 days, including clinical symptoms, mortality, changes in body weight, and macroscopic examination of the liver and kidneys. Results: No mortality or clinical signs of toxicity were observed in any group. Body weight increased normally with no significant differences (p > 0.05). Macroscopic examination showed normal liver and kidney features (color, size, consistency). Histopathological evaluation (H&E staining) revealed no significant abnormalities: renal glomeruli and tubules normal (degeneration score 0 in all samples); liver with no hepatocyte necrosis (score 0). LD₅₀ > 15 g/kg BW. Conclusion: The combination of citronella and neem did not cause any mortality or acute toxic symptoms. this material is classified as non-toxic in acute exposure (LD₅₀ > 15 g/kg BW) and safe for veterinary herbal formulation development.