A healthy vaginal environment is predominated by certain Lactobacillus species, which lead to the prevention of infections of the reproductive tract. This study examined the characteristics of cultivable Lactobacillus species in both healthy women and women with bacterial vaginosis (BV). Between November 2011 and September 2013, 139 women attending a women's clinic in Mysore, India, were evaluated for BV in a cross-sectional study. BV was diagnosed using Amsel's criteria: homogeneous vaginal discharge, vaginal pH >4.5, production of amines, and presence of "clue" cells. Those with three or more of the characteristics were considered to have BV. Vaginal swabs were then cultured in Rogosa agar and de Man-Rogosa-Sharpe broth. Gram-positive lactobacilli generating 600-800 bp amplicons by 16 sRNA were further characterized by sequencing. Cultivable vaginal samples were obtained from 132 women (94.9%). According to the Amsel criteria, 83 women (62.1%) were healthy, and 49 (37.1%) had BV. Eleven different Lactobacillus species were isolated from 47 women. The common lactobacilli species found in this sample included L. crispatus (39.6%), L. gasseri (45.8%), and L. jensenii (14.6%). Lactobacilli were isolated from 39 healthy women and eight with BV. L. gasseri was cultured from 18.8% of healthy women and 6.1% with BV. The presence of L. reuteri was significantly associated with normal vaginal microbiota (P-value = 0.026). These results further our understanding of vaginal lactobacilli colonization and richness in this particular population. Our findings showed that lactobacilli species present in the vaginas of healthy women in India do not differ from those reported from other countries.