The Efficacy of Pasteurised Akkermansia Muciniphila in Healthy Medical Workers

Study Overview
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Brief Summary
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Gut microbiota alterations secondary to chronic stress might serve as a triggering factor towards manifestation of somatic and mental symptoms. The administration of pasteurised A. muciniphila MucT has the capability of supporting microbiota and improving the gut barrier integrity, which might lead to decrease of inflammation and the negative health consequences of stress in healthy participants.

Detailed Description
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The Gut-brain-microbiota axis (GBMA) is a bi-directional pathway, both neuronal and biochemical, between the intestine and the Central Nervous System (CNS). The gut microbiota plays a central role in gut-brain communication. The composition of intestinal microbiota and its functions play an important role in the pathogenesis of disorders of gut-brain interaction - both within the digestive tract and in the brain. Modulation of gut microbiota with the aid of probiotics, antibiotics, or germ-free feeding protocols significantly altered stressful event-induced behavioral outcomes in rodents. Moreover, the intake of various probiotics significantly improved stress-induced anxiety and depressive-like behaviors in mice. In humans, probiotics were also documented to display some beneficial effects on mental health, including alteration of emotional bias in healthy individuals, and alleviating stress and anxiety among stressed adults. Psychobiotics are imposed with certain limitations related to their standardization and end-shelf-life product stability. Therefore, the use of postbiotics, which contain bacterial metabolites or other bacteria derived fragments are viewed as novel solutions and alternatives to use of standard probiotics. One of novel postbiotics of interest among scientists and clinicians is pasteurized Akkermansia muciniphila MucT (PAM). Animal studies indicate that administration of Akkermansia muciniphila can ameliorate metabolic syndrome, obesity, diabetes, and inflammatory bowel disease in animals and has psychobiotic potential. Similar to live A. muciniphila, PAM could ameliorate several diseases as well. The mechanism of action of PAM - improving gut barrier integrity - suggests the potential use to reduce the negative effects of stress. Human studies shown that PAM is safety, what was confirmed in the Scientific Opinion of EFSA. Recently A. muciniphila was approved as the Novel Food. A proof of concept study will be conducted to verify the hypothesis that PAM reduces the psychological and somatic effects of stress.

Official Title
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The Effects of the Anti-inflammatory Microbe - Pasteurized Akkermansia Muciniphila (PAM) on Symptoms of Somatic and Mental Stress in Healthcare Professionals

Conditions
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Healthy, Stress, Dietary Supplement

Intervention / Treatment
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* Dietary Supplement: Pasteurized Akkermansia muciniphila
* Dietary Supplement: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Working in a high stress hospital department, like: emergency, trauma, intensive care, surgery, internal diseases; Written informed consent to participate in this study before any study-mandated procedure; Body mass index (BMI) ≥18.5 kg/m2 and ≤ 35 kg/m2; A willingness and motivation to follow the study protocol. Exclusion Criteria: Diagnosis of autoimmune, neurological, immunocompromised, thyroid, inflammatory bowel diseases, irritable bowel syndrome, diabetes, cancer, and/or IgE-dependent allergy; Psychiatric comorbidities, including mental retardation, organic brain dysfunction, or addiction (except nicotine and caffeine), intake of antipsychotic and antidepressive drugs; Proton pump inhibitors usage; The use of antibiotics and/or probiotics 4 weeks prior to the study; Glucocorticosteroids and/or metformin treatment; Dietary supplementation (except for vitamin D) within the three months before screening; Specific restrictive (e.g. elimination, vegan, FODMAP, reduction) diet within the three months before screening; Significant changes in physical activity 4 weeks before the trial entry; Pregnancy or lactation; Significant GI surgery within the last 6 months prior to or planned during the study; Any other medication for management of IBS complaints like peppermint oil, bile acid binders; Lactose intolerance; Participation in another study during the last 30 days prior to and during the study; Any other reason for exclusion as per investigator's judgment, e.g. insufficient compliance with study procedures.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: parallel-groups, randomized, placebo-controlled clinical study
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Pasteurized Akkermansia muciniphila<br>pasteurized A.muciniphila (Pasteurized, 3x10^10 bacteria per day) as supplement for 3 months, | Dietary Supplement: Pasteurized Akkermansia muciniphila<br>* PAM supplementation; packaging will be given to the subjects every one month during follow-up visits, with the instructions to take one dose every morning on an empty stomach<br>* Other names: PAM;|
| Placebo Comparator: Placebo<br>placebo administered for 3 months once a day | Dietary Supplement: Placebo<br>* PBO administration; packaging will be given to the subjects every one month during follow-up visits, with the instructions to take one dose every morning on an empty stomach<br>* Other names: PBO;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Stress intensity | serum dehydroepiandrosterone sulfate (DHEAS) in blood | baseline | 
| Stress intensity | serum dehydroepiandrosterone sulfate (DHEAS) in blood | 1 month | 
| Stress intensity | serum dehydroepiandrosterone sulfate (DHEAS) in blood | 3 months | 
| Cardiovascular marker of stressor intensity | blood pressure | baseline | 
| Cardiovascular marker of stressor intensity | blood pressure | 1 month | 
| Cardiovascular marker of stressor intensity | blood pressure | 3 months | 
| Cardiovascular marker of stressor intensity | heart rate | baseline | 
| Cardiovascular marker of stressor intensity | heart rate | 1 month | 
| Cardiovascular marker of stressor intensity | heart rate | 3 months | 
| Stress intensity | Perceived Stress Scale (PSS-10). Individual scores on the PSS can range from 0 to 40 with higher scores indicating higher perceived stress. | baseline | 
| Stress intensity | Perceived Stress Scale (PSS-10). Individual scores on the PSS can range from 0 to 40 with higher scores indicating higher perceived stress. | 1 month | 
| Stress intensity | Perceived Stress Scale (PSS-10). Individual scores on the PSS can range from 0 to 40 with higher scores indicating higher perceived stress. | 3 months | 
| Psychosocial working conditions | Copenhagen Psychosocial Questionnaire (COPSOQ). The scales of the COPSOQ are formed by adding the points of the individual questions of the scales by giving equal weights to each question. In most cases the questions have five response options. In these cases the weights are: 0, 25, 50, 75, and 100. The scale value is calculated as the simple average | baseline | 
| Psychosocial working conditions | Copenhagen Psychosocial Questionnaire (COPSOQ). The scales of the COPSOQ are formed by adding the points of the individual questions of the scales by giving equal weights to each question. In most cases the questions have five response options. In these cases the weights are: 0, 25, 50, 75, and 100. The scale value is calculated as the simple average | 1 month | 
| Psychosocial working conditions | Copenhagen Psychosocial Questionnaire (COPSOQ). The scales of the COPSOQ are formed by adding the points of the individual questions of the scales by giving equal weights to each question. In most cases the questions have five response options. In these cases the weights are: 0, 25, 50, 75, and 100. The scale value is calculated as the simple average | 3 months | 
| The recognition of the most common mental disorders | Primary Care Evaluation of Mental Disorders (PRIME-MD). The yes/no questionnaire serves as an initial screen for 5 general groups of mental disorders commonly found in the general population | baseline | 
| The recognition of the most common mental disorders | Primary Care Evaluation of Mental Disorders (PRIME-MD). The yes/no questionnaire serves as an initial screen for 5 general groups of mental disorders commonly found in the general population | 1 month | 
| The recognition of the most common mental disorders | Primary Care Evaluation of Mental Disorders (PRIME-MD). The yes/no questionnaire serves as an initial screen for 5 general groups of mental disorders commonly found in the general population | 3 months | 
| Depression | Patient Health Questionnaire-9 (PHQ-9). Nine items, each of which is scored 0 to 3, providing a 0 to 27 severity score.This score can then be referred to the accompanying PHQ-9 Scoring Box to interpret the TOTAL score. | baseline | 
| Depression | Patient Health Questionnaire-9 (PHQ-9). Nine items, each of which is scored 0 to 3, providing a 0 to 27 severity score.This score can then be referred to the accompanying PHQ-9 Scoring Box to interpret the TOTAL score. | 1 month | 
| Depression | Patient Health Questionnaire-9 (PHQ-9). Nine items, each of which is scored 0 to 3, providing a 0 to 27 severity score.This score can then be referred to the accompanying PHQ-9 Scoring Box to interpret the TOTAL score. | 3 months | 
| Depressive state | The Beck Depression Inventory (BDI). When the test is scored, a value of 0 to 3 is assigned for each answer and then the total score is compared to a key to determine the depression's severity. The standard cut-off scores were as follows: 0-18: indicates minimal depression 18-30: indicates mild depression 19-29: indicates moderate depression 30-63: indicates severe depression. | baseline | 
| Depressive state | The Beck Depression Inventory (BDI). When the test is scored, a value of 0 to 3 is assigned for each answer and then the total score is compared to a key to determine the depression's severity. The standard cut-off scores were as follows: 0-18: indicates minimal depression 18-30: indicates mild depression 19-29: indicates moderate depression 30-63: indicates severe depression. | 1 month | 
| Depressive state | The Beck Depression Inventory (BDI). When the test is scored, a value of 0 to 3 is assigned for each answer and then the total score is compared to a key to determine the depression's severity. The standard cut-off scores were as follows: 0-18: indicates minimal depression 18-30: indicates mild depression 19-29: indicates moderate depression 30-63: indicates severe depression. | 3 months | 
| Anxiety and stress | Depression Anxiety Stress Scale 21 (DASS-21). This is a set of three self-report scales designed to measure the emotional states of depression, anxiety and stress. The rating scale is as follows: 0 - Did not apply to me at all - Applied to me to some degree, or some of the time - Applied to me to a considerable degree, or a good part of time - Applied to me very much, or most of the time. SUBSCALES: DASS_Anxiety = questions 2 + 4 + 7 + 9 + 15 + 19 + 20 DASS_Depression = questions 3 + 5 + 10 + 13 + 16 + 17 + 21 DASS_Stress = questions 1 + 6 + 8 + 11 + 12 + 14 +18 | baseline | 
| Anxiety and stress | Depression Anxiety Stress Scale 21 (DASS-21). This is a set of three self-report scales designed to measure the emotional states of depression, anxiety and stress. The rating scale is as follows: 0 - Did not apply to me at all - Applied to me to some degree, or some of the time - Applied to me to a considerable degree, or a good part of time - Applied to me very much, or most of the time. SUBSCALES: DASS_Anxiety = questions 2 + 4 + 7 + 9 + 15 + 19 + 20 DASS_Depression = questions 3 + 5 + 10 + 13 + 16 + 17 + 21 DASS_Stress = questions 1 + 6 + 8 + 11 + 12 + 14 +18 | 1 month | 
| Anxiety and stress | Depression Anxiety Stress Scale 21 (DASS-21). This is a set of three self-report scales designed to measure the emotional states of depression, anxiety and stress. The rating scale is as follows: 0 - Did not apply to me at all - Applied to me to some degree, or some of the time - Applied to me to a considerable degree, or a good part of time - Applied to me very much, or most of the time. SUBSCALES: DASS_Anxiety = questions 2 + 4 + 7 + 9 + 15 + 19 + 20 DASS_Depression = questions 3 + 5 + 10 + 13 + 16 + 17 + 21 DASS_Stress = questions 1 + 6 + 8 + 11 + 12 + 14 +18 | 3 months | 
| Occurrence of Irritable Bowel Syndrome | Rome IV criteria | baseline | 
| Occurrence of Irritable Bowel Syndrome | Rome IV criteria | 1 month | 
| Occurrence of Irritable Bowel Syndrome | Rome IV criteria | 3 months | 
| Occurence and severity of gastrointestinal symptoms | Gastrointestinal Symptom Rating Scale (GSRS) | baseline | 
| Occurence and severity of gastrointestinal symptoms | Gastrointestinal Symptom Rating Scale (GSRS) | 1 month | 
| Occurence and severity of gastrointestinal symptoms | Gastrointestinal Symptom Rating Scale (GSRS) | 3 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Microbiota composition | next generation sequencing | baseline | 
| Microbiota composition | next generation sequencing | 1 month | 
| Microbiota composition | next generation sequencing | 3 months | 
| A. muciniphila count in stool | real-time quantitative PCR (qPCR) | baseline | 
| A. muciniphila count in stool | real-time quantitative PCR (qPCR) | 1 month | 
| A. muciniphila count in stool | real-time quantitative PCR (qPCR) | 3 months | 
| Total bacteria count in stool | real-time quantitative PCR (qPCR) | baseline | 
| Total bacteria count in stool | real-time quantitative PCR (qPCR) | 1 month | 
| Total bacteria count in stool | real-time quantitative PCR (qPCR) | 3 months | 
| Short chain fatty acids content in stool | quadrupole mass spectrometer and high performance liquid chromatograph | baseline | 
| Short chain fatty acids content in stool | quadrupole mass spectrometer and high performance liquid chromatograph | 1 month | 
| Short chain fatty acids content in stool | quadrupole mass spectrometer and high performance liquid chromatograph | 3 months | 
| Immune phenotypes of peripheral blood mononuclear cells (PBMCs) | single-cell genomics (scRNA-seq and scATAC-seq) analyses (in blood) | baseline | 
| Immune phenotypes of peripheral blood mononuclear cells (PBMCs) | single-cell genomics (scRNA-seq and scATAC-seq) analyses (in blood) | 1 month | 
| Immune phenotypes of peripheral blood mononuclear cells (PBMCs) | single-cell genomics (scRNA-seq and scATAC-seq) analyses (in blood) | 3 months | 
| Inflammatory mediators concentrations in blood | high-throughput protein biomarker analysis with the advent of Proximity Extension Assay | baseline | 
| Inflammatory mediators concentrations in blood | high-throughput protein biomarker analysis with the advent of Proximity Extension Assay | 1 month | 
| Inflammatory mediators concentrations in blood | high-throughput protein biomarker analysis with the advent of Proximity Extension Assay | 3 months | 
| Zonulin concentration in stool | enzyme-linked immunosorbent assay (ELISA) | baseline | 
| Zonulin concentration in stool | enzyme-linked immunosorbent assay (ELISA) | 1 month | 
| Zonulin concentration in stool | enzyme-linked immunosorbent assay (ELISA) | 3 months | 
| Calprotectin concentration in stool | enzyme-linked immunosorbent assay (ELISA) | baseline | 
| Calprotectin concentration in stool | enzyme-linked immunosorbent assay (ELISA) | 1 month | 
| Calprotectin concentration in stool | enzyme-linked immunosorbent assay (ELISA) | 3 months | 
| Lipopolysaccharide concentration in blood | enzyme-linked immunosorbent assay (ELISA) | baseline | 
| Lipopolysaccharide concentration in blood | enzyme-linked immunosorbent assay (ELISA) | 1 month | 
| Lipopolysaccharide concentration in blood | enzyme-linked immunosorbent assay (ELISA) | 3 months | 
| Adiposity | Fat mass/fat free mass evaluated by bioimpedance | baseline | 
| Adiposity | Fat mass/fat free mass evaluated by bioimpedance | 1 month | 
| Adiposity | Fat mass/fat free mass evaluated by bioimpedance | 3 months | 
| Obesity | Body weight | baseline | 
| Obesity | Body weight | 1 month | 
| Obesity | Body weight | 3 months | 
| Dietary habits | the frequency of certain food consumption (rank score) by means of validated Food Frequency Questionnaire (FFQ). | baseline | 
| Physical activity | International Physical Activity Questionnaire. Results can be reported in categories (low activity levels, moderate activity levels or high activity levels) or as a continuous variable (MET minutes a week). | baseline | 
| Functions of peripheral blood mononuclear cells (PBMCs) | mass cytometry (CyTOF) | baseline | 
| Functions of peripheral blood mononuclear cells (PBMCs) | mass cytometry (CyTOF) | 1 month | 
| Functions of peripheral blood mononuclear cells (PBMCs) | mass cytometry (CyTOF) | 3 months | 
| Insulin resistance | HOMA-Homeostasis Model Assessment calculated from fasted glycemia and insulinemia | baseline | 
| Insulin resistance | HOMA-Homeostasis Model Assessment calculated from fasted glycemia and insulinemia | 1 month | 
| Insulin resistance | HOMA-Homeostasis Model Assessment calculated from fasted glycemia and insulinemia | 3 months | 
| carbohydrate metabolism | glycated hemoglobin (HbA1c) | baseline | 
| carbohydrate metabolism | glycated hemoglobin (HbA1c) | 1 month | 
| carbohydrate metabolism | glycated hemoglobin (HbA1c) | 3 months | 
| Concentration of blood lipids | Analysis of circulating lipids : total, LDL and HDL cholesterol (mg/dl), triglycerides (md/dl) | baseline | 
| Concentration of blood lipids | Analysis of circulating lipids : total, LDL and HDL cholesterol (mg/dl), triglycerides (md/dl) | 1 month | 
| Concentration of blood lipids | Analysis of circulating lipids : total, LDL and HDL cholesterol (mg/dl), triglycerides (md/dl) | 3 months | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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microbiota, postbiotic, stress, metabolome, immunometabolism