Aerobic vaginitis (AV) is a vaginal infection caused mainly by overgrowth of Escherichia coli and a reduction of Lactobacilli in the vagina. The infection is linked with adverse pregnancy outcomes, such as preterm birth. The current medical therapy for AV with antibiotics is associated with adverse effects and necessitates the use of alternative treatments. Apple cider vinegar (ACV) is a natural fermented product produced from apples and is reported to exhibit potent antibacterial activity. ACV also contains lactic acid bacteria, which can act as probiotics. Symptoms of AV can be improved by restoring the disturbed microbial imbalance rather than exposing the vagina to synthetic drugs. Hence, an attempt was made to investigate whether ACV could support growth of the beneficial bacteria and inhibit growth of the pathogenic bacteria such as E. coli in the vagina. In vitro evaluation of the anti-bacterial potential of ACV against E. coli showed a potent antibacterial activity, recording a zone of inhibition diameter of 32.9 ± 0.5 mm; however, no zone of inhibition was observed against Lactobacillus acidophilus. Turbidimetric analysis was used to ensure growth of the bacteria in a simulated vaginal fluid (SVF) by using a nephelometer as a Nephelometric Turbidity Unit (NTU). Both E. coli and L. acidophilus were grown individually in SVF containing ACV. The ACV expressed antibacterial efficacy against E. coli and favoured growth of L. acidophilus in the SVF. Conclusively, ACV can help in growth of the beneficial bacteria and restore the natural microbiota of the vagina, thus proving to be beneficial in the management of AV.