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Abstract. Food companies show a major concern for developing novel foods to expand their market among which bakery products are an emerging category. Biscuit with its attractive features surges the demand for new and health improved products for probiotic delivery.To date, very few studies on biscuits containing viable microorganisms haveexisted. This is due to a challenge for bakery products employed with high temperatures during baking. Therefore, in the present work, a biscuit wasmade from dough with an addition of free probiotic cells of strain Lactobacillus acidophilusto study the influence of baking conditions on its survival. The viable cell counts, temperature profile, moisture content, and water activity of the biscuit were determined after baking for 0, 4, 8, and 12 min at 180 and 200°C, respectively. The reduction of probiotic cells during the entire baking was about 5 logs for 180 °C.Biscuits baked at 180 and 200 °C were found to have a final moisture content of 3.74% and 1.5% respectively.