Vinegars are commonly used as food condiments and preservatives. Apple cider vinegar (ACV) is also used in the Ayurvedic pharmaceutical industry because of its medicinal properties. Since specifically selected starter cultures for commercial vinegar production are not readily available, apple juice supplemented with sugar is commonly inoculated with a microbiologically undefined culture obtained from the previous batch of ACV. The present work focuses on the isolation of yeasts and acetic acid bacteria from ACV and the preparation of a starter culture. ACV was produced in a bench scale bioreactor using a traditional fermentation process wherein an acetic acid concentration of 3.8% was obtained after three weeks. Several acetic acid bacteria (AAB) were isolated from ACV using selective media. Microscopy revealed the cultures to be gram negative to gram variable short rods. The growth pattern of the isolates on differential media and biochemical tests suggested the presence of Acetobacter and Gluconobacter species. Ten potent isolates were selected for starter culture preparation. Two consortia were formulated with five AAB isolates in each along with a yeast isolate and used for ACV production, wherein an acetic acid concentration of 4.2%-4.9% was obtained in 10-12 days. Thus, these two starter cultures with locally isolated AAB can be used for the commercial production of apple cider vinegar.