Milk Ingredients and Resistance Against E-coli-induced GastroEnteritis (MIRAGE)

Study Overview
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Brief Summary
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Background: The incidence of gastrointestinal infections is very high. In Western countries at least 30% of the population suffers from at least one food-borne infection per year. Mostly because of the problem of antibiotic resistance, more emphasis is put on prevention of infections. One of the possibilities is to strengthen human resistance to gut infections by consumption of milk ingredients. Aim: To study whether a milk protein concentrate rich in phospholipids improves the resistance of humans to enterotoxigenic E. coli (ETEC). Study design: The MIRAGE study is a parallel, double-blind, placebo-controlled 4-weeks intervention with a milk protein concentrate rich in phospholipids in healthy subjects of 18-55 yrs of age. Participants will be randomly assigned to the milk protein concentrate rich in phospholipids or placebo group (n=30 per group). Subjects will be instructed to maintain their usual pattern of physical activity and their habitual food intake, but to standardize their dietary calcium intake. After an adaptation period of 2 weeks, subjects will be orally infected with a live, but attenuated, ETEC vaccine (strain E1392-75-2A; collection NIZO food research; dose will be 1010 CFU). Before and after infection, an online diary will be kept to record all food and drinks consumption (2x2 days) to assess the habitual dietary intake. The diary will also be used for daily recording of bowel habits and frequency and severity of gastrointestinal complaints. The following biological samples will be collected: 4x10 ml venous blood, a single fecal bolus (for screening) and 7x24 hrs feces. Blood is sampled for immune response analyses and the fecal samples are collected to quantify several infection- and immune system markers and to verify dietary calcium intake. Saliva is sampled three times before and after infection to quantify immune system markers. Primary outcomes: Fecal ETEC excretion and severity of diarrhea (quantified by fecal output per day). Secondary outcomes: Serum immune response to ETEC, self-reported stool consistency scores and gastrointestinal complaints, relative fecal wet weight.

Official Title
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A Randomized, Placebo-controlled, Double Blind Volunteer Study Into the Effect of Milk Ingredients on Gastroenteritis Caused by an Attenuated E.Coli.

Conditions
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Bacterial Infection, Diarrhea, Gastroenteritis

Intervention / Treatment
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* Dietary Supplement: Milk protein
* Dietary Supplement: Milk protein rich in phospholipids


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Signed informed consent Age 18-55 yrs Availability of internet connection Willingness to replace habitual dairy product intake with the supplied low-calcium soy products Willingness to abstain from products with high amounts of prebiotic fibers and from products with probiotics starting 1 month prior to study start Willingness to give up blood donation from 1 month before the start of the experiment and during the entire experimental period. Exclusion Criteria: Current or previous underlying disease of the GI tract, liver, bile bladder, kidney, thyroid gland (self-reported) Allergy to milk products or lactose intolerance (self-reported), since the capsules may contain milk traces from culture media Allergy to soy products (self-reported) Use of antibiotics, norit, laxatives (up till 6 months prior to inclusion), cholestyramine, acid burn inhibitors or immune suppressive agents (up till 3 months prior to inclusion), and pre- and probiotics (up till 1 month prior to inclusion). High titer serum antibodies against ETEC (10 ml blood sample collected at screening) ETEC detected in fecal sample (collected at screening) Vegetarians Vegans Heavy alcohol use (>4 consumptions/day or >20/week) Drug use

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 55 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: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Milk protein<br>Milk powder, twice daily at breakfast and dinner. | Dietary Supplement: Milk protein<br> <br> |
| Experimental: Milk protein rich in phospholipids<br>Milk powder, twice daily at breakfast and dinner. | Dietary Supplement: Milk protein rich in phospholipids<br> <br> |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change of fecal ETEC numbers over time as a marker of intestinal colonization resistance | The change over time in fecal ETEC numbers is compared between treatment and placebo group. | Days -1 and -2 before ETEC challenge and days 1,2,3,4,7 and 15 after challenge | 
| Change of total daily fecal output over time as a marker of infectious diarrhea | The change in the daily fecal output over time is compared between treatment and placebo group. | Days -1 and -2 before ETEC challenge and on days 1,2,3,4,7 and 15 after challenge. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Daily Bristol Stool Score as a marker for stool consistency |  | Days -1 and -2 before ETEC challenge and on days 1-15 after challenge. | 
| Gastro-Intestinal Symptom Rating Scale (GSRS) for gastro-intestinal complaints |  | Days -1 and -2 before ETEC challenge and on days 1-15 after challenge | 
| % fecal wet weight as a marker for Diarrhea severity |  | Days -1 and -2 before ETEC challenge and days 1-15 after challenge. | 
| Stool frequency (number of stools per day) | Days -1 and -2 before ETEC challenge and on days 1-15 after challenge | Day -1 and -2 and days 0-15 | 
| Specific serum antibody response to CFAII |  | Before ETEC challenge and on day 15 after challenge. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Bacterial infection, Dairy, Diarrhea, Diet, E.Coli, Gastroenteritis, Milk, Stool consistency