Probiotics, Brain Structure and Psychological Variables

Study Overview
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Brief Summary
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Theory and research on the gut-brain-axis emphasize complex interactions between the gut microbiota, immunological and hormonal responses, brain function, brain structure, as well as resulting behavioral manifestations, such as cognitive functions and mental illness. Probiotics are living micro-organisms that change the composition of the gut microbiota and hypothetically have a positive effect on the host's general health and well-being. Probiotic bacteria naturally occur in foods such as Sauerkraut, olives, and dark chocolate, and are currently also added to industrial products such as yogurt. Regarding the effect of probiotics on brain structure and function, animal studies have shown that the administration of probiotics in mice and rats was linked to neurogenesis in the hippocampus and an improvement of associated cognitive functions. The majority of these studies applied probiotics for 4 weeks. The substances used in these studies were often composed of several bacterial strains, suggesting that the neurogenic effect may not be reducible to a specific type of probiotic bacteria. Probiotics seem to be effective in improving memory abilities, including spatial and non-spatial memory, both in rodents and humans. Moreover, specifically regarding the beneficial effect of probiotics on anxiety, depression and stress, preliminary evidence in humans is compelling. However methodologically sound (randomized-controlled trial [RCT], 'blind') studies are still lacking. To sum up, the present study is going to be the first RCT with human participants that investigates structural and functional changes of the hippocampus through probiotic bacteria, using Magnet Resonance Imaging (MRI). In addition, the study aims at advancing research in the field by investigating the effects of probiotics on a broad spectrum of cognitive functions, particularly those associated with hippocampal activity (e.g. spatial memory, verbal memory). Furthermore, effects on several types of mental illness (e.g. anxiety, depression, stress) will be studied. Potential translatory mechanisms that may promote the aforementioned effects will be examined, i.e. changes in immunological parameters, 'brain derived neurotrophic factor' (BDNF), and oxytocin.

Detailed Description
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A double-blind RCT intervention study will be conducted to investigate the effect of a probiotic dietary supplement vs. a placebo, taken by healthy human participants (N = 60) for 28 consecutive days. Both substances are administered in doses of 4.4g (powder) per day. The probiotic dietary supplement used in the present study is a freely available product (in powder form) sold under the name of Vivomixx®. Per 4.4g the powder contains 450 billion bacteria, composed of eight bacterial strains: Lactobacilli (L. paracasei, L. plantarum, L. acidophilus and L.delbrueckii subsp. bulgaricus), Bifidobacteria (B. longum, B. infantis, B. breve), and Streptococcus thermophiles. This product was chosen, because a similar supplement with an identical composition of bacteria as Vivomixx® was found to be linked to hippocampal neurogenesis in mice. The placebo powder has a similar taste and consistency as the verum (Vivomixx®) powder, but contains no probiotic bacteria. First, participants will be screened online for eligibility (T0). Before and after the 28-day intake period, participants are asked to perform several cognitive tests and fill out questionnaires (T1, T2) at our laboratory (ca. 2 - 2.5 hours). Afterwards, blood samples will be drawn (ca. 5 minutes) and a magnet resonance tomography (MRT) session (ca. 1 - 1.5 hours) will be conducted, where participants will perform an object pattern separation task in the scanner (T1, T2). Furthermore, a follow-up after 2 months (T3), including the same assessments as at T1 and T2, will be conducted. During the intake period participants will fill out a brief questionnaire on a weekly basis, which includes an assessment of state depression and anxiety, defecation (control item to rule out adverse effects), recent (start of) intake of medication, recent (onset of) intake of antibiotics, and recent inter-current illness (e.g. influenza). In addition, participants will record their food intake (protocol) for one week (7 consecutive days) during the intake period.

Official Title
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Effects of Probiotic Bacteria on Brain Structure and Function and on Psychological Variables

Conditions
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Depressive Symptoms, Anxiety, Stress

Intervention / Treatment
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* Dietary Supplement: Vivomixx® powder
* Other: Placebo powder


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: healthy participants age 18-40 informed consent for all parts of the study (including MRT) none of the exclusion criteria (see below) fulfilled Exclusion Criteria: age < 18 or > 40 years pregnancy or breastfeeding left-handedness degenerative or inflammatory diseases of the central nervous system severe cognitive/ neuropsychological impairment severe pain syndrome or other severe organic diseases epilepsy (past or present) psychiatric disorders neurological disorder severe diabetic polyneuropathy malignancies/ cancer cardiac insufficiency arterial hypertension heart attack/ stroke severe hepatic or renal insufficiency diseases of the hemopoietic system alcoholism/ drug addiction medical history of severe allergic or toxic reactions current participation in drug trial doubts about legal capacity/ capability of understanding referral to institutions based on court/ official order treatment with centrally acting medication (e.g. antipsychotics, antiepileptics, antidepressants, etc.) non-removable metal pieces (aneurysm clips, artificial limbs, etc.) or implanted electronic devices (pacemaker, osmotic or other implanted pumps, cochlear implants, etc.) claustrophobia acute (respiratory) infection, physical uneasiness tattoos in the head region, permanent make-up non-removable piercings chronic/ acute gastrointestinal diseases vegetarianism, veganism current intake of antibiotics or intake of antibiotics at any time point during the last 2 months lactose intolerance general focus on probiotic diet

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 40 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: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: 1 Verum group, receiving the probiotic Vivomixx® powder 1 Placebo group, receiving a placebo powder (similar taste and consistency as Vivomixx® powder)
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: verum condition probiotics<br>The verum condition probiotics in the present study is a freely available product, Vivomixx® powder (dietary supplement). Each dose (4.4g) contains 450 billion bacteria, composed of eight bacterial strains: Lactobacilli (L. paracasei, L. plantarum, L. acidophilus, L.delbrueckii subsp. bulgaricus), Bifidobacteria (B. longum, B. infantis, B. breve), and Streptococcus thermophiles. 30 Participants will be randomly assigned to this condition. The intake period is 28 days, daily dose = 4.4g. | Dietary Supplement: Vivomixx® powder<br>* Participants will take in a daily dose of 4.4g for 28 consecutive days<br>* Other names: probiotic dietary supplement;|
| Placebo Comparator: placebo condition<br>In the placebo condition participants will receive a placebo powder (comparable in taste and consistency to Vivomixx® = verum condition probiotics) that contains no probiotic bacteria. 30 Participants will be randomly assigned to that condition. The intake period is 28 days, daily dose = 4.4g. | Other: Placebo powder<br>* Participants will take in a daily dose of 4.4g for 28 consecutive days<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in hippocampal volume, assessed via Magnet Resonance Imaging (MRI) | changes in hippocampal volume in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in functional brain activation during fMRI task | changes in functional connectivity (using BOLD signal) in hippocampal regions in verum experimental group (in comparison to placebo control) during pattern separation fMRI task | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in depression | changes in levels of depression, assessed with Beck's Depression Inventory - II Revised (BDI-II-R) sum score in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in spatial navigation | changes in test performance scores (number of correct responses, degree of accuracy measured as position hits) in the Tunnel task in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in Interleukin-6 (IL-6) | changes in IL-6 blood serum concentration levels in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in IL-1ß | changes in IL-1ß blood serum concentration levels in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in Tumor Necrosis Factor alpha (TNF-alpha) | changes in TNF-alpha blood serum concentration levels in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in Brain Derived Neurotrophic Factor (BDNF) | changes in blood serum level concentration of BDNF in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in verbal learning test performance | changes in verbal learning performance score, assessed with the Verbal Learning Memory Test (VLMT) in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 
| Changes in pattern separation fMRI task | changes in the pattern separation task performance (no. of correct responses to picture stimuli) in in verum experimental group (in comparison to placebo control) | at baseline (day 0) after intake period (day 28+) and at follow up (day 84+) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Probiotics, Psychiatric Symptoms, Cognitive Function, Hippocampus, Oxytocin, Immunological Parameters