Application of Vaginal-spraying LiveSpo X-secret in Supporting Treatment of Sexually Transmitted Diseases in Vietnamese Women

Sexually Transmitted Infections (STIs) are common gynecological infections worldwide, especially in tropical and underdeveloped countries where up to three-quarters of the population has STIs, mainly in the reproductive age. The infection rate in regions and countries is different, the infection rate in developing countries is 20-fold higher than in developed countries. Southeast Asia alone has about 150.5 million people with these diseases. Vietnam is located in the tropics, and hot and humid climates are favorable factors for microbial pathogens to develop. According to reports from the dermatology units of the provinces, every year about 300,000 people are infected with sexually transmitted diseases...There are many microbial pathogens causing infections including bacteria, viruses, fungi, and protozoa...The patient can be infected with multiple pathogens at the same time. This is a matter of great concern in public health care because STIs often have mild or no symptoms, so they are easily overlooked. While antibiotics or antifungal drugs are typically used to treat STIs caused by bacterial or fungal agents like Chlamydia, Gonorrhea, and Candida, no specific treatment exists for most viral STIs. However, the use of broad-spectrum antibiotics can result in side effects like loss of appetite, fatigue, and sexual problems, while repeated treatments can lead to the emergence of antibiotic resistance, posing a significant challenge for STI treatment worldwide. Probiotics are microorganisms that are beneficial to our health when consumed in adequate amounts, playing a vital role in maintaining a healthy balance of bacteria in our body, especially in the gut and vaginal areas. Studies have found that probiotics can help reduce the incidence and severity of vaginal infections, including bacterial vaginosis and yeast infections. Currently, oral or vaginal suppository probiotics containing strains of Bifidobacterium or Lactobacillus are being studied as a potential complementary therapy for treating vaginal infections. However, these strains have low survivability and are sensitive to light, moisture, and pH levels, making preservation challenging. Thus, their effectiveness depends on the specific strains used and the dose required to achieve colonization in the vaginal tract. Bacillus is a well-studied probiotic strain with over 350,000 research publications worldwide, and its safety has been extensively investigated. Bacillus subtilis, B. clausii, and B. coagulans are all aerobic or facultative anaerobic bacteria capable of spore formation and can tolerate harsh conditions such as nutrient deprivation, anoxia, drought, and even antibiotics. Here, investigators propose that vaginal-spraying probiotics containing Bacillus strains could be a safe and effective symptomatic treatment for STIs caused by viral, fungi, or bacterial pathogens, providing a promising alternative to traditional antibiotic therapy.

The objective was to investigate the symptomatic treatment effects of the probiotic product LiveSpo X-secret as a liquid-suspension form containing Bacillus spores of safe B. subtilis ANA4, B. clausii ANA39, B. coagulans ANA 40 strains, in women having sexually transmitted infection and investigators evaluation of improved efficacy and reduced clinical symptoms treatment time and measurement of changes in pathogen load in the vaginal samples before treatment (day 0) and after 3, 7, 14 and 28 days using LiveSpo X-secret.

Methods: A randomized, blind, and controlled clinical trial is conducted. After informed consent, patients will be screened for common nine STI pathogens in Vietnam including Gardnerella vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis, Mycoplasma genitalium, Human alphaherpesvirus type 1 and 2, Candida albicans, Trichomonas vaginalis. Totally 100 patients positive with at least one of nine STIs will be randomized into 2 groups (n = 50/group): the control group (named "Control" group) uses 0.9% NaCl physiological saline and an experimental group (named the "X-secret" group) use the probiotics LiveSpo X-secret. The patient is given a coded spray in the form of a blind sample to ensure the objectivity of the study. The clinical follow-up will be at the time point of 0, 3, 7, 14, and 28 days, vaginal samples will be collected at days 0, 3, 7, 14, and day 28 to evaluate potential reductions in pathogens load and change the microbiome in vaginal flora.

Real-time PCR for detection of microorganism in vaginal samples: semi-quantitative assays for measuring changes in pathogens load is conducted by the real-time PCR assay which has been optimized and developed by the research team (Bui et al., 2023). Detection of useful Bacillus strains belonging to B. subtilis, B. clausii, and B. coagulans is conducted by real-time PCR SYBR Green assay that is routinely runs also conducted at Spobiotic Research Center, Hanoi, Vietnam

Analysis of vaginal microbiome 16S rRNA using next-generation sequencing (NGS) at Macrogene (Seoul, Korea) on the Illumina MiSeq (Illumina, San Diego, CA, USA) using a 2 × 250 bp run configuration.