Accurate 16S Absolute Quantification Sequencing Revealed Vaginal Microecological Composition and Dynamics During Mixed Vaginitis Treatment With Fufang FuRong Effervescent Suppository

Abstract

Background: The diagnosis and treatment of mixed vaginitis are more complicated than single pathogenic infections, and there may be adverse reactions and several contraindications to conventional antibiotic therapy. Therefore, this study aimed to evaluate the preliminary effects of Fufang Furong Effervescent Suppository for the management of aerobic vaginitis (AV) mixed with bacterial vaginosis (BV) using Accurate 16S absolute quantification sequencing (Accu16S).

Methods: In the present randomized, blind, multi-center clinical trial, women (20 to 55 years) who had received a diagnosis of AV+BV were randomly assigned into clindamycin positive control (n = 41) and Fufang Furong Effervescent Suppository (n = 39) groups. The follow-up occurred in three time periods (V1: -2~0 days; V2: 15-17 days; V3: 40 ± 3 days). At each visit, two vaginal swabs, one for clinical evaluation and one for laboratory examination, were taken from each patient. The Nugent score, Donders' score, drug-related complications, recurrence rates, and microecological changes of vaginal swabs were assessed in the time three periods.

Results: At baseline, the two groups were similar in frequency of presentation with vaginal burning, odor, abnormal discharge, and itching. No meaningful differences in Nugent and Donders' scores were detected between the two groups at stage V2 (Nugent: p = 0.67; Donders': p = 0.85) and V3 (Nugent: p = 0.97; Donders: p = 0.55). The Furong group presented fewer complications compared to the Clindamycin group. However, this difference was not statistically significant (p = 0.15). Additionally, Accu16S indicated that the total abundance of bacteria in both groups sharply decreased in stage V2, but slightly increased in V3. In stage V3, the absolute abundance of Lactobacillus in the Furong group was considerably higher compared to untreated samples (p < 0.05). On the other hand, no momentous increase was detected in the Clindamycin group (p > 0.05).

Conclusion: Fufang Furong Effervescent Suppository can be as effective as clindamycin cream in the management of AV+BV while may restore the vagina microecosystem better.

Keywords: accurate 16S absolute quantification sequencing; antibiotic; microbiome; mixed vaginitis; traditional Chinese medicine.

Figure 1

Nugent score (A) and Donders score (B) before and after Clindamycin and Furong treatments. (****P value < 0.0001).

Figure 2

Flowchart of Accu16S qualified sample screening.

Figure 3

Comparisons of the vagina microbiota α diversity (A, B) and relative (C) and absolute (D) abundances in the principal coordinate analysis (PCoA) of bacterial β diversity. (A, B) rank all the mean values from highest to lowest using significant difference letter notation. The largest mean is marked with the letter a and compared with other means, where the difference is not significant, the letter a is marked until an average with a significant difference is marked with the letter b.

Figure 4

Relative and absolute abundances of the major taxa at the genus (A) and the species (B) levels in Clindamycin and Furong groups.(Ta, Tb, and Tc represent stages V1, V2, and V3 of the Furong group, respectively; Ca, Cb, and Cc represent stages V1, V2, and V3 of the Clindamycin group, respectively).

Figure 5

The changes of dominant microorganisms between the two groups at each visit: Gardnerella (A), Prevotella (B), Atopobium (C), Sneathia (D), Megasphaera (E), and Lactobacillus (F). (Ta, Tb, and Tc represent stages V1, V2, and V3 of the Furong group, respectively; Ca, Cb, and Cc represent stages V1, V2, and V3 of the Clindamycin group, respectively; ns, nonsense; *0.01≤P value < 0.05; **0.001≤P value < 0.01; *** 0.0001≤P value < 0.001; ****P value < 0.0001).

Figure 6

Heatmap of the distributions of the 17 most abundant bacterial genera present in the vaginal swab samples. (The abscissa represents the sample, the ordinate represents different species at the genus level. The absolute abundances of bacterial genera are indicated by color intensity. Ta, Tb, and Tc represent stages V1, V2, and V3 of the Furong group, respectively; Ca, Cb, and Cc represent stages V1, V2, and V3 of the Clindamycin group, respectively).