The invention discloses insect and acarus killing shampoo for animals and a preparation method thereof. The shampoo is prepared from sodium lauryl ether sulfate, coconut oil momoethanolamide, cocoamido propyl hydroxy sulfobetaine, C12-13 alkyl lactate, natural essence, vegetable amino acids, deionized water and at least one of azadirachtin or neem oil, and is further prepared from at least one of pyrethrin, cnidium lactone and sophocarpidine. The preparation method comprises the steps of weighing and dispersing. According to the formula, the azadirachtin and the neem oil are adopted as main functional components and are reasonably compatible with other raw materials, so that symptoms such as animal skin itching, local erythema and unhairing can be effectively relieved. The insect and acarus killing shampoo is safe, environmentally friendly, good in insect and acarus control effect and lasting in control effect and is prepared from simple components. According to the preparation method, a nanometer shearing machine is adopted for shearing raw materials, the raw materials are sufficiently mixed to be uniform through the nanometer technology, the preparation method is simple, convenient to implement, low in cost and applicable to large-scale industrial production.