Vaginitis is defined as a spectrum of conditions that cause vaginal and sometimes vulvar symptoms, such as itching, burning, irritation, odor, and vaginal discharges. This study aims to identify and isolate the most prevalent causal agents associated with vaginitis at Gezira area – Sudan, evaluating the antimicrobial activity of the commonly used herbs like Commiphora myrrha, Solenostemma argel, Azadirachta indica, Salvia officinalis and Eucalyptus camaldulensis against the isolated causal agents and comparing the herbal effects at different concentrations with a known commercial vaginal washer. 50 subjects of high vaginal swabs were collected, to identify and isolate the target organisms during the period from January to May 2015, and then aseptically cultured on sterile and appropriate media. A total number of 31 bacterial isolates were represented by 4 different species of bacteria; 17 (34%) Staphylococcus aureus, 7 (14%) E. coli, 6(12%) Streptococcus pyogenes and only 1(2%) Pseudomonas aeruginosa whereas 14 (28%) yeast isolates were represented by Candida albicans. 11 aqueous extracts were prepared from each of the five herbs with concentrations of 5%, 15%, 25%, 35%, and 45%. The aqueous extracts of C. myrrha, E. camaldulensis and A. indica respectively achieved the highest means of inhibition zones (12.1, 11.5 and 10.4 mm) when compared to the others and showing non-significant differences when compared to each other (p value 0.08, 0.2 and 0.7). The combination of (S. argel, A. indica, and E. camaldulensis) showed relatively strong antimicrobial activity with inhibition zone range between (18.5–24 mm) in its different concentrations (25%, 35%, and 45%) when comparing the effect of this extract (mean=15.2) with the impact of the commercial washer (mean=9.9), it was found that the herbal extract has top influence, although, the difference was statistically non-significant (p value ± 0.07).