Oxaliplatin, an important drug for the treatment of colorectal cancer, often produces neuropathic cold allodynia even after a single administration. Bee venom acupuncture (BVA) has been traditionally used in Korea to treat various pain symptoms and is shown to have a potent anti-allodynic effect in nerve injured animals. Our previous study also demonstrated that BVA alleviates oxaliplatin-induced cold allodynia, which is mediated partially by the noradrenergic system. This study further investigated whether the serotonergic and cholinergic systems mediate the anti-allodynic effect of BVA in oxaliplatin-injected rats. The behavioral signs of cold allodynia induced by an oxaliplatin injection (6 mg/kg, i.p.) were evaluated by a tail immersion test in cold water (4°C). The significant allodynia was observed from 3 days after an oxaliplatin injection. The established allodynia was significantly attenuated by BVA treatment (0.25 mg/kg, s.c.) at GV3 acupoint, Yoyanggwan. Depletion of serotonin by pretreatment of DL-pchlorophenylalanine (PCPA, 150 mg/kg/day, i.p., 3 days) abolished such anti-allodynic effect of BVA. In addition, either of methysergide (mixed 5-HT1/5-HT2 receptor antagonist, 1 mg/kg, i.p.) or MDL-72222 (5-HT3 receptor antagonist, 1mg/kg, i.p) blocked the BVA-induced anti-allodynia. We also found that mecamylamine (non-selective nicotinic antagonist, 2mg/kg, i.p.), but not atropine (non-selective muscarinic antagonist, 1 mg/kg, i.p.), prevented the BVA effect. Further, Dihydro-&beta-erythroidine hydrobromide (Dh&betaE, &alpha4&beta2 nicotinic antagonist, 5 mg/kg, i.p.), but not methyllycaconitine citrate (MLA, &alpha7 nicotinic antagonist, 6 mg/kg, i.p.) blocked the anti-allodynic effect of BVA. Taken together, these results suggest that BVA alleviates oxaliplatin-induced acute cold allodynia in rats, at least in part, through the activation of the serotonergic and cholinergic systems, especially 5-HT1-3 receptors and &alpha4&beta2 nicotinic receptors.