The Effect of Probiotics on E. Coli Gastroenteritis (PRETEC)

Background:

The incidence of gastrointestinal infections is very high. In European countries 10-25% of the population suffers from at least one foodborne infection per year. Probiotics may strengthen human resistance to gut infections as they may beneficially modulate the intestinal microbiota composition and activity, and the immune function upon intestinal infection.

Aim:

To study whether probiotics improve the resistance of humans to enterotoxigenic E. coli (ETEC).

Study design:

The PRETEC study is a parallel, double-blind, placebo-controled 4-weeks intervention with probiotics in healthy volunteers. In this study, the effect of probiotic intervention vs placebo on several infection markers in response to an ETEC challenge is investigated. Participants will be randomly assigned to the probiotic or placebo group (n=21 per group). Subjects will be instructed to maintain 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 10E10 CFU). This ETEC strain induces mild and short-lived infectious diarrhea symptoms. Before and after infection, a diary will be kept to write down 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. Blood is sampled for immune response analyses and multiple faecal samples are collected to quantify several infection- and immune system markers, to determine probiotic excretion, and to verify dietary calcium intake.

Study population:

Healthy males of 20-55 yrs of age.

Interventions:

Probiotics (freeze-dried powder, dose 10E9 CFU twice daily) or placebo (carrier material powder of identical appearance).

Primary outcomes:

Total fecal ETEC excretion per day and severity of diarrhea (quantified by faecal output per day).

Secondary outcomes:

Serum immune response to ETEC, self-reported stool consistency scores and gastrointestinal complaints, relative faecal wet weight, sIgA and calprotectin in faeces, probiotic persistence and levels of opportunistic pathogens in the endogenous microbiota.

Study Overview

• Status: Completed
• Conditions: Gastroenteritis; Diarrhea; Bacterial Infection
• Intervention / Treatment: Dietary supplement: probiotics; Dietary supplement: placebo

Detailed Description

The timeframes for analysis of the primary and secondary outcomes is mentioned below in the Outcome Measures section.

• Study Type: Interventional
• Enrollment (Actual): 40
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Netherlands
  • Ede, Netherlands, 6718 ZB
    • NIZO food research

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 20 years to 55 years (Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: Male

Description

Inclusion Criteria:

• Signed informed consent
• Male
• Age 20-55 yrs
• Willingness to replace habitual dairy product intake with the supplied low- calcium soy milk products
• Willingness to abstain from products with high amounts of prebiotic fibers and products with probiotics (except for the supplied one)

Exclusion Criteria:

• Current or previous underlying disease of the GI tract
• lactose intolerance
• Use of antibiotics, norit, laxatives, cholestyramine, acid burn inhibitors, immune suppressiva, prebiotics, probiotics
• detectable serum antibodies against ETEC
• carriage of streptomycin-resistant E. coli in faeces (only relevant when culturing technique for ETEC will be applied instead of specific RT-PCR)
• vegetarians
• heavy alcohol use
• drug use

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Prevention
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Triple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: probiotics; Freeze-dried powder, dose 10E9 CFU twice daily for 4 weeks	Dietary supplement: probiotics; Freeze-dried powder, dose 10E9 CFU twice daily for 4 weeks
Placebo Comparator: placebo; Carrier material powder of identical appearance	Dietary supplement: placebo; placebo consisting of carrier material powder of identical appearance

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Time Frame
Total daily faecal ETEC excretion with time	1-2 days before ETEC infection and on days 1, 2, 3, 5 and 15 after ETEC infection
Total daily faecal output with time	1-2 days before ETEC infection and on days 1, 2, 3, 5, 15 after ETEC infection

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Bowel habits		Scored daily in a diary
Frequency and severity of gastrointestinal symptoms		Scored daily by VAS scales in a diary
Diarrhea severity	Determined as % faecal wet weight	1-2 days before ETEC infection and on days 1, 2, 3, 5, and 15 after ETEC infection
Opportunistic pathogens in faeces		At a single day just before ETEC infection and at a single time point in the first week after ETEC infection
Faecal calprotectin		1-2 days before ETEC infection and on days 1, 2, 3, 5, and 15 after ETEC infection
Serum antibody response to CFA II		At screening (baseline) and at days 9 and 15 after ETEC infection
Total faecal sIgA		1-2 days before ETEC infection and on days 1, 2, 3, 5, and 15 after ETEC infection

Collaborators and Investigators

• Sponsor: NIZO Food Research
• Collaborators: Danisco
• Investigators: Principal Investigator: Sandra ten Bruggencate, PhD, NIZO food research; Principal Investigator: Ingeborg Bovee-Oudenhoven, PhD, NIZO food research; Principal Investigator: Arthur Ouwehand, PhD, Danisco Finland

Publications and helpful links

General Publications

• Ouwehand AC, ten Bruggencate SJ, Schonewille AJ, Alhoniemi E, Forssten SD, Bovee-Oudenhoven IM. Lactobacillus acidophilus supplementation in human subjects and their resistance to enterotoxigenic Escherichia coli infection. Br J Nutr. 2014 Feb;111(3):465-73. doi: 10.1017/S0007114513002547. Epub 2013 Aug 12.

Study record dates

Study Major Dates

• Study Start: October 1, 2010
• Primary Completion (Actual): December 1, 2010
• Study Completion (Actual): April 1, 2011

Study Registration Dates

• First Submitted: October 15, 2010
• First Submitted That Met QC Criteria: October 19, 2010
• First Posted (Estimate): October 20, 2010

Study Record Updates

• Last Update Posted (Estimate): August 27, 2015
• Last Update Submitted That Met QC Criteria: August 26, 2015
• Last Verified: August 1, 2015

More Information

Terms related to this study

• Keywords: probiotics; prevention; dietary intervention; intestinal infection; E. coli infection; infectious diarrhea; intestinal resistance
• Additional Relevant MeSH Terms: Digestive System Diseases; Signs and Symptoms, Digestive; Gastrointestinal Diseases; Bacterial Infections and Mycoses; Infections; Diarrhea; Gastroenteritis; Bacterial Infections
• Other Study ID Numbers: N132716.081.10