BACKGROUND: Reducing the protein and amino acid content of diet or altering the dietary electrolyte balance were suggested as the effective strategies to prevent the harmful effects of the heat stress in poultry.OBJECTIVES: This study surveyed the effects of various levels of threonine and dietary cation-anion balance on the broiler chickens under the heat stress conditions.METHODS: 700 one-day-old male broilers (Ross 308) were used for 6 weeks under high- temperature conditions. This experiment was conducted using a 2 × 3 + 1 factorial experiment based on a completely randomized design, with 7 treatments (5 replicates per treatment and 20 birds per replicate). Experimental factors included two threonine concentrations (recommended level and 10 % above recommended level) and three dietary electrolyte balances (175, 250, and 325 mEq/kg). The control group considered the treatment with a normal threonine concentration (recommended level) and a dietary electrolyte balance of 250 mEq/kg under optimal temperature conditions.RESULTS: Although there was no interaction between threonine level and electrolyte balance, the electrolyte balance of 175 meq/kg increased the performance index compared to the electrolyte balance of 250 or 325 mEq/kg (P<0.05). The villus height to crypt depth ratio and villus surface area in the jejunum improved with an electrolyte balance of 175 or 250 mEq/kg (P<0.05). A decrease in the dietary electrolyte balance increased in the nitrogen-corrected apparent metabolizable energy (P<0.05). Besides, an increase in the population of lactobacilli and bifidobacteria in the cecum, as well as an increase in protein digestibility, were observed by increasing levels of threonine in the diet (P<0.05).CONCLUSIONS: Based on these results, a high level of threonine or a low electrolyte balance in diet (175 mEq/kg) may improve the performance, metabolizable energy, and intestinal microbial flora of broilers reared under heat stress.