Four types of traditional Thai rice—polished, black fragrant, glutinous and black glutinous rice—were separately used as raw material for vinegar production. During alcohol fermentation, using enriched baker’s dried yeast (S. cerevisiae) as a starter culture gave the highest ethanol content over 7 days of fermentation. The conversion of ethanol to acetic acid for vinegar production by Acetobacter pasteurianus TISTR 102 was performed for 25 days. The highest amount of acetic acid was detected with glutinous rice fermentation (6.68% w/v). The biological properties of Thai rice vinegars were determined, including the total phenolic content, antioxidant activity, antibacterial activity and cytotoxicity. Black glutinous rice vinegar exhibited the maximum total phenolic content of 133.68 mg GAE/100 mL. This result was related to the antioxidative activity findings, for which black glutinous rice vinegar exhibited the strongest activity against both ABTS^•+ and DPPH^• radicals. Cytotoxicity against the human colon cancer cell line (HT-29) provided an IC₅₀ value of 74.02 μg/mL and weak activity in a mouse fibroblast normal cell line (L929) with an IC₅₀ value of 171.06 μg/mL. Glutinous rice vinegar was the most effective vinegar for inhibiting pathogenic bacterial growth of both Gram-negative and Gram-positive bacteria. These results suggested that the value of total phenolic content corresponded to the anticancer activity and antioxidant activity results, while antibacterial activity depended on the acidity of rice vinegar.