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Abstract The main objectives of this work aimed to investigate the acute and short-term toxicity of two neonicotinoids pesticides, acetamiprid and thiamethoxam, that are widely used in Egypt and South Africa in agriculture, veterinary, and household pest control in albino Swiss mice. These data are needed as most of the farmers is exposed to low doses of pesticides through residues in food products and beverages. Hence, the study was undertaken to investigate the sub-acute toxicity of orally administered pesticide for 28 days. Swiss albino mice males were used for the acute and sub-acute (sub-chronic) tests. Neonicotinoids are a new generation of widely-used insecticides. For their outstanding potency and systemic action, they are used for crop protection against pest insects in agriculture and domestic insect control. The neonicotinoids act as agonists to the nicotinic acetylcholine receptors (nAChRs) in the central nervous system of insects with greater affinity than to mammals. However, their residues are present in soil, dust or water and might still become a health problem for animals and humans. Thousands of tons of neonicotinoids are widely used around the world as broad-spectrum systemic insecticides and veterinary drugs. In 2014, neonicotinoids accounted for more than 25% of the global insecticide market. and about 20,000 tons of the active substance imidacloprid was produced in 2010). Therefore, neonicotinoids are now being considered for replacement of many existing conventional insecticide classes (e |