Establishing Effects of Bacillus Clausii Probiotic Consumption on Gastrointestinal Health and Inflammation

Prospective participants are pre-qualified by email screening and attended an in-person study visit at Colorado State University's Food and Nutrition Clinical Research Laboratory (FNCRL) in the Department of Food Science and Human Nutrition. During the initial visit, participants signed the Consent Form after the nature of the procedure had been fully explained to them. Their eligibility is confirmed by taking anthropometric measurements of height (cm) and weight (kg) to calculate BMI and by completing a written medical health questionnaire to determine medical history and current medication use. Eligible participants are then assigned sequentially to a study code (MS01, MS02, etc.), and codes were pre-assigned to intervention sequences using a random number generator in Microsoft Excel. For the initial clinic visit (and subsequent visits), participants are fasted for at least 8 hours and asked to abstain from alcohol use for at least 12 hours prior to clinic visits. They were also instructed to refrain from taking any non-prescription medications for 24 hours or dietary supplements for 12 hours before their visits. At the initial visit, a venous blood sample was collected, and participants filled out several assessment forms/questionnaires: GI Health Appraisal Questionnaire, GI Symptom Rating Scale (GSRS), GI Quality of Life Questionnaire (IBS-QOL Questionnaire) 7-day Physical Activity Recall, Positive and Negative Affect Schedule, and Perceived Stress Scale.

• Bacillus (Shouchella) clausii CSI08 (Dietary supplement), also known as Munispore: Gram-positive, spore-forming microorganism
• Placebo (Other): Inert filler material delivered in a capsule identical to the probiotic intervention

Primary outcomes

• Adverse Event Rate [12 weeks]: For each arm adverse event forms were tracked and the total rate of adverse events was calculated
• Gastrointestinal symptom rating score [12 weeks]: Gastrointestinal symptoms were assessed using the gastrointestinal symptom rating score, at the beginning and end of each intervention period. The GSRS is a 15-item scale where each item can be scored 1-7 according to symptom severity. A minimum score is 15 and the maximum score is 110. The higher score indicates more severe symptoms.

Secondary outcomes

• Gut microbiota composition [12 weeks]: The gut microbiota composition was assessed by 16s rRNA at the beginning and end of each intervention period.
• Systemic Inflammation [12 weeks]: PBMCs were collected at the beginning and end of each intervention period and then cultured and stimulated with bacterial LPS. Collected supernatants were assessed using a multiplex ELISA for T-cell activity.
• Regularity of Bowel Movements [13 weeks]: A Bristol stool scale was used to record daily bowel movements throughout the study to determine changes in bowel movement regularity.

Other outcomes

• Blood Lipids [12 weeks]: Blood lipids were assessed using collected whole blood samples at the beginning and end of each intervention period using a Piccolo Xpress Blood Lipid Panel.
• Liver Toxicity [12 weeks]: A comprehensive metabolic panel will be performed on whole blood collected at the beginning and end of each intervention period using a Piccolo Xpress Comprehensive Metabolic Panel. This includes multiple markers, that when considered together, can provide an indication of liver toxicity. Specifically, out-of-range values for alanine amino transferase (ALT), asparagine amino transferase (AST), alkaline phosphatase (AST), bilirubin, albumin, and total protein can collectively be interpreted to look for indication of hepatocellular or cholestatic injury.

Sex: All, ages: 18 Years to 75 Years, accepts healthy volunteers

Inclusion Criteria:

* Age 18-75
* BMI 18-34.9
* self report as healthy
* consistent Mild to Moderate gastrointestinal distress (determined by a functional GI screen)

Exclusion Criteria:

* <18 or >75
* BMI <18 or >34.9
* antibiotics within two months
* probiotic/prebiotic supplements within two weeks prior to the screening process
* pregnant or breastfeeding
* taking medications that influence study endpoints of the study (i.e., statins, metformin, nonsteroidal anti-inflammatory drugs, and monoamine oxidase inhibitors)
* a current diagnosis of cancer, liver or kidney disease, metabolic disorders, and GI diseases (Celiac disease, inflammatory bowel disease, and peptic ulcer disease)

• Tiffany L Weir, PhD, Principal investigator, CSU
• Allegra L Vazquez, MS, Study director, CSU