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Acerola (Malpighia emarginata) vinegar is one kind of fruit vinegar produced using acetic acid fermented by acetic acid bacteria. In this study, Acetobacter senegalensis A28 was applied in acerola vinegar production at a high temperature. With 1.0% (v/v) of A. senegalensis A28 inoculum, the suitable conditions for production of acerola vinegar were determined to be pH 6.5, 20oBrix, 106 cells/ml, as well as 4.0% (v/v) of ethanol added to acerola juice. Moreover, a temperature of 35oC was found to be suitable for fermentation in both 100 ml and 2-litre working volumes. Total acid concentrations of 5.45% (w/v) and 4.90% (w/v), respectively, were achieved after 4 days of fermentation. Acid concentrations of 5.40% (w/v) and 4.70% (w/v) were obtained at 37oC after 5 days of fermentation in volumes of 100 ml and 2 litres, respectively. At pilot-scale fermentation of 20 litres at 37oC, acid concentration of 3.68% (w/v) was achieved after 5 days of fermentation.