Promising clinical and immunological efficacy of Bacillus clausii spore probiotics for supportive treatment of persistent diarrhea in children

Abstract

Persistent diarrhea is a severe gastroenteric disease with relatively high risk of pediatric mortality in developing countries. We conducted a randomized, double-blind, controlled clinical trial to evaluate the efficacy of liquid-form Bacillus clausii spore probiotics (LiveSpo CLAUSY; 2 billion CFU/5 mL ampoule) at high dosages of 4-6 ampoules a day in supporting treatment of children with persistent diarrhea. Our findings showed that B. clausii spores significantly improved treatment outcomes, resulting in a 2-day shorter recovery period (p < 0.05) and a 1.5-1.6 folds greater efficacy in reducing diarrhea symptoms, such as high frequency of bowel movement of ≥ 3 stools a day, presence of fecal mucus, and diapered infant stool scale types 4-5B. LiveSpo CLAUSY supportive treatment achieved 3 days (p < 0.0001) faster recovery from diarrhea disease, with 1.6-fold improved treatment efficacy. At day 5 of treatment, a significant decrease in blood levels of pro-inflammatory cytokines TNF-α, IL-17, and IL-23 by 3.24% (p = 0.0409), 29.76% (p = 0.0001), and 10.87% (p = 0.0036), respectively, was observed in the Clausy group. Simultaneously, there was a significant 37.97% decrease (p = 0.0326) in the excreted IgA in stool at day 5 in the Clausy group. Overall, the clinical study demonstrates the efficacy of B. clausii spores (LiveSpo CLAUSY) as an effective symptomatic treatment and immunomodulatory agent for persistent diarrhea in children.Trial registration: NCT05812820.

Keywords: Bacillus clausii; Cytokine; Persistent diarrhea; Probiotic; Spore.

Figure 1

A flowchart illustrating the participant enrolment process in this study. Participants were initially screened for eligibility using the clinical database. Those who met the criteria and provided informed consent were then randomly assigned to either the Control or Clausy group. Data collection and analysis were carried out between day 0 (baseline) and the end of the treatment period. The study recruited participants and conducted clinical and subclinical measurements from July 2022 to July 2023.

Figure 2

Days of treatment (A1–C1) for observation of three typical symptoms of diarrhea and time-dependent percentage (%) of asymptomatic patients (A2–C2) in the Control (dashed lines) and Clausy (solid lines) groups. Methods to test distribution was verified by Mann–Whitney test or t-test.

Figure 3

Improvement in duration and efficacy of treatment to resolve persistent diarrhea by probiotic intervention. (A) Overall treatment days to resolve persistent diarrhea. (B) time-dependent percentage (%) of asymptomatic patients (B) in the Control (dashed lines) and Clausy (solid lines) groups. (C) Kaplan–Meier analysis of estimated median time to resolve the persistent diarrhea in the Control (dashed lines) and Clausy (solid lines) groups. (D) Total duration of diarrhea from the onset of symptoms to the complete recovery from the disease. Methods to test distribution in Fig. 4A, D were verified by Mann–Whitney test.

Figure 4

Pro- and anti-inflammatory cytokines levels (pg/mL) in blood samples and IgA levels (μg/mL) in stool samples of Control and Clausy groups at day 5 compared to day 0. The Wilcoxon test was used to calculate the median differences in IL-17 (A), IL-23 (B), IL-6 (C), and TNF-α (D) pro-inflammatory cytokine, anti-inflammatory IL-10 (E) cytokine, and IgA (F) levels at day 0 and day 5 in each group. The Mann–Whitney test was used to compared cytokine and IgA concentrations (A–F) between the two groups. Only samples with measurable cytokine and IgA concentrations at day 0 were included in the statistical analysis. 95% CI for median in each group and the median difference between the two groups were shown in this figure. The significance level of all analyses was set at the p < 0.05.