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Yang and Sihn: The Growth Inhibition Effect on the Bacterial Vaginosis Causative Bacteria by Citric Acid and Trisodium Phosphate
Original Article

J Bacteriol Virol 2015;45(3):228-234.

Published online: 9 February 2015

DOI: https://doi.org/10.4167/jbv.2015.45.3.228

The Growth Inhibition Effect on the Bacterial Vaginosis Causative Bacteria by Citric Acid and Trisodium Phosphate

Nam-Woong Yang1, ∗, Wan-Jin Sihn2

1Department of Microbiology, College of Medicine, Chosun University, Gwangju, Korea

2Department of Alternative Complementary Medicine, Graduate School Chosun University, Gwangju, Korea

∗Corresponding author: Nam-Woong Yang. Department of Microbiology, College of Medicine, Chosun University, 309 Pilmun-daero Dong-gu Gwangju 501-759, Korea. Phone: +82-62-230-6350, Fax: +82-62-232-3125, e-mail: nwyang@chosun.ac.kr

∗∗ This study was supported by research fund from Chosun University, 2014.

Received 30 May 201516 July 2015       Accepted 27 July 2015

Copyright © 2015 Journal of Bacteriology and Virology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Bacterial vaginosis (BV) is the most frequent vaginal disease being apt to relapse. The growth inhibition effect of the mixture of citric acid (CA) and trisodium phosphate (TSP) on BV causative bacteria and probiotics was measured. Gardnerella vaginalis was reduced to zero in WCCT-1 (CA 0.25% and TSP 0.55% in Wilkins-Chalgren broth), 2.0 × 104/ml in WCCT-2 (CA 0.5% and TSP 0.8% in WC), and 3.3 × 103/ml in WCCT-3 (CA 1.0% and TSP 2.6% in WC) comparing with 1.3 × 105/ml in WC after 48 h. Bacteroides fragilis was reduced to 6.0 × 103/ml in WCCA (CA 0.34% in WC), 2.3 × 102/ml in WCCT (CA 0.5% and TSP 0.2% in WC), 7.0 × 103/ml in WCHCl (HCl in WC) after 48 h. Mobiluncus mulieris was reduced to 1.08 × 104/ml in WCCA, 1.03 × 103/ml in WCCT, and 10 ea/ml in WCHCl after 48 h. Peptostreptococcus asaccharolyticus was completely inhibited in WCCA, WCCT, and WCHCl after 24 h. Probiotics, Steroidobacter denitrificans YH1 (3.4 × 107/ml) and Lactobacillus crispatus YH2 (2.7 × 106/ml), grew to 1.25 × 108/ml and 2.6 × 107/ml in MRSCA (CA 1.0% in MRS), 1.8 × 107/ml and 4.6 × 106/ml in MRSCT (CA 1.5% and TSP 0.58% in MRS), 1.2 × 108/ml and 2.3 × 107/ml in MRSHCl after 48 h, respectively. These results mean that the CA-TSP mixture can be used as the useful vaginal pH controller, growth inhibitor on BV causative bacteria, and an efficient means for settlement of probiotics.

Keywords: Bacterial vaginosis, Citric acid, Trisodium phosphate, Probiotics

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Table 1.
Growth rate of G. vaginalis according to media supplements
Media\hr 241 48
WCCT-12 6.0 × 105/ml NG
WCCT-23 4.2 × 105/ml 2.0 × 104/ml
WCCT-34 4.2 × 105/ml 3.3 × 103/ml
Control5 1.29 × 108/ml 1.3 × 105/ml

1. Counts of inittial bacterial inoculation are 2.3 × 106/ml

2. WC broth conntaining 0.25% CA and 0.55% TSP, pH 6.27

3. WC broth conntaining 0.5% CA and 0.8%% TSP, pH 6.27

4. WC broth conntaining 1% CA and 2.6% TTSP, pH 6.27

5. WC broth conntrol, pH 6.92

Table 2.
Growth rate of B. fragilis according to media suppleme
Media/hr 241 48
WCCA2 1.6 × 105/ml 6.0 × 103/ml
WCCT3 1.3 × 104/ml 2.3 × 102/ml
WCHCl4 6.2 × 105/ml 7.0 × 103/ml
Control5 2.2 × 109/ml 1.4 × 109/ml

1. Counts of initial bacterial inoculation are 7.0 × 108/ml

2. WC broth containing 0.34% CA, pH 4.2

3. WC broth containing 0.5% CA and 0.2% TSP, pH 4.2

4. WC broth controlled by 5 N HCl to pH 4.2

5. WC broth control, pH 6.92

Table 3.
Growth rate of M. mulieris according to media sup plements
Media/hr 241 48
WCCA2 7.6 × 104/ml 1.08 × 104/ml
WCCT3 1.5 × 103/ml 1.03 × 103/ml
WCHCl4 30 ea/ml 10 ea/ml
Control5 3.6 × 108/ml 4.6 × 109/ml

1. Counts of initial bacterial inoculation are 3.2 × 108/ml

2. WC broth containing 0.34% CA, pH 4.2

3. WC broth containing 0.5% CA and 0.2% TSP, pH 4.

4. WC broth controlled by 5 N HCl to pH 4.2

5. WC broth control, pH 6.92

Table 4.
Growth rate of P. asaccharolyti supplements
Media/hr 241 48
WCCA3 NG2 NG
WCCT4 NG NG
WCHCl5 NG NG
Control6 5.5 × 106/ml 1.0 × 107/ml

1. Amounts of initial bacterial inoculation are 3.0 × 106/ml

2. No growth

3. WC broth containing 0.34% CA, pH 4.2

4. WC broth containing 0.5% CA and 0.2% TSP, pH 4.2

5. WC broth controlled by 5 N HCl to pH 4.2

6. WC broth control, pH 6.92

Table 5.
Growth supplements rate of S. denitrificans YH1 according to media
Media/hr 241 48
MRSCA2 6.8 × 106/ml 1.25 × 108/ml
MRSCT3 6.7 × 105/ml 1.8 × 107/ml
MRSHCl4 1.2 × 108/ml 1.2 × 108/ml
MRS5 1.9 × 108/ml 2.0 × 108/ml

1. Counts of initial bacterial inoculation are 3.4 × 107/ml

2. MRS broth containing 1% CA, pH 4.2

3. MRS broth containing 1.5% CA and 0.6% TSP pH 4.2

4. MRS broth controlled to pH 4.2 by 5 N HCl

5. MRS broth control, pH 5.72

Table 6.
Growth rate of L. crispatus YH2 according to media supplements
Media/hr 241 48
MRSCA2 2.5 × 103/ml 2.6 × 107/ml
MRSCT3 3.5 × 105/ml 4.6 × 106/ml
MRSHCl4 1.33 × 106/ml 2.3 × 107/ml
MRS5 3.0 × 107/ml 7.0 × 106/ml

1. Amounts of initial bacterial inoculation are 2.7 × 106/ml

2. MRS broth containing 1% CA, pH 4.2

3. MRS broth containing 1.5% CA and 0.6% TSP pH 4.2

4. MRS broth controlled by 5 N HCl to pH 4.2

5. MRS broth control, pH 5.7

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