Combined Lactobacilli (Reuteri LMG P-27481 and GG ATCC 53103) to Prevent Antibiotic Associated Symptoms in Children
Combined Lactobacilli (Reuteri LMG P-27481 and GG ATCC 53103) to Prevent Antibiotic Associated Symptoms in Children
This is a prospective multicenter pediatric double-blind, randomized placebo-controlled,
 parallel group clinical trial intended to evaluate the efficacy, safety and tolerability of a
 combined probiotic (LGG and L. reuteri LMG P-27481) in the prevention of antibiotic
 associated diarrhea and gastrointestinal symptoms in hospitalised children treated with
 antibiotics for proven or suspected bacterial infection.
Eligible children will be randomly assigned to receive the combined probiotic study product
 (Noos, Roma, Italia)(BCCMTM Bacterial Collection of Ghent, Belgium, Italian Patent:
 102016000011071, International Patent Request : PCT/IB 2017/053856), or placebo as additional
 treatment to the antibiotic therapy. Kind, dose and duration of antibiotic treatment will be
 decided by the local clinicians according to good medical practice and current
 recommendations. The first dose of probiotic will be started the same day of starting the
 antibiotic treatment, immediately after enrollment. Then, each oral stick will be
 administered orally once daily, in the morning, before breakfast, during the antibiotic
 treatment and prolonged for 30 days since starting.

 Study product The combined probiotic is made by Lactobacillus reuteri LMG P-27481 and
 Lactobacillus rhamnosus GG (ATCC 53103), sucralose and isomalt as an oral stick formulation
 of 1.6 g with a concentration of probiotic of 2x1010 colony forming unit (CFU) per stick,
 launched in the Italian market with the name of Reuterin®GG (Noos, Roma, Italia).

 The probiotic and placebo will be prepared centrally by the probiotic producer as identically
 appearing oral stick. The placebo will contain only sucralose and isomalt, 1.6 g per stick.

 Randomization The allocation sequences and randomization lists has been generated by a online
 computer system. The probiotic -placebo allocation ratio will be of 1:1. To ensure allocation
 concealment, an independent subject prepared the randomization schedule and sealed envelopes
 to be sent to the participant centers. Each investigator will control the packaging and
 labels of trial treatments sent by the producer with a numerical code with no possible
 identification of the probiotic product compared to the placebo. The allocation will be
 consecutively assigned for all enrolled subjects. All investigators, participants, and
 statisticians will be blinded to the assigned treatment throughout the study and during the
 data analysis. After completing the analysis of the data or in case of sever adverse event
 the randomization code will be broken and information about the content of the product
 (probiotic or placebo) will be revealed by the producer, upon the investigator's request.

 Monitoring and collection of data At enrollment a standardized form will record age, gender,
 anthropometric parameters, vital signs, refilling time, site of infection, laboratory tests,
 kind, dose and route of administration of antibiotic, diet, previous AAD or other
 gastrointestinal symptoms, comorbidity and associated treatment. Vesikary scale,
 anthropometric measures, vital signs, refilling time will be also recorded in each patient at
 enrollment and in case of occurrence of diarrhea. Bristol stool chart will be used to grade
 the consistency of stools.

 Parents or care givers of each children will be instructed to fill in a diary to record the
 duration of antibiotic treatment and/or other treatment, the intake of the study product, the
 frequency of daily bowel movements, the consistency of stools (according to the Bristol stool
 scale), the frequency and intensity (according to a VAS scale, from 0 to 10) of abdominal
 pain, the presence and duration of fever, vomiting or any other symptoms they considered
 important, diet changes and general well being (according to a numerical 1-10 scale). The
 diary will be completed daily for the duration of the probiotic treatment and then weekly
 till the end of the study. After discharge from the hospital the absence from day care or
 school, the need of new antibiotic treatment or re-hospitalization will be also noted. In
 case of diarrhea, fever or other severe symptoms after hospital discharge parents will be
 instructed to contact the local pediatrician and investigator for clinical evaluation and
 stool analysis. A clinical visit will be scheduled in all recruited patients after 1, 3 and 6
 months from enrollment.

 Re-hospitalization will be required in case of severe infection, severe dehydration or
 whenever considered necessary by clinicians.

 Scheduled investigations In all cases presenting diarrhea at least three stool samples will
 be collected and stool tests will be performed to detect viral, bacterial or C. difficile
 pathogens. Blood and urine test or radiologial investigations will be decided by the local
 clinician based on patient's clinical presentation and according to guidelines and good
 clinical practice.

 One additional sample of stools will be collected, whenever possible, in each patient at
 enrollment, at 1, 3 and 6 months to evaluate fecal calprotectin to indirectly assess
 intestinal inflammation. In a subgroup of patients (the first 30 recruited in each arms) two
 other stool samples and a urine sample will be collected and stocked in freezer (-20° C) at
 the same study point for possible future additional explorative analysis of fecal
 inflammatory markers (lactoferrine, IgA, IL10 and TNF-α) and metabolomic profile (urine
 metabolites).

 The probiotic producer (Noos S.r.L, Roma, Italia) will gift the probiotic product and placebo
 for the duration of the study for all children enrolled but has and will have no role in the
 design and conduct of the study; collection, management, analysis, or interpretation of the
 data; preparation, review, or approval of the manuscript; or the decision to submit the
 manuscript for publication. The data of the enrolled subjects will be managed, stored and
 property of the clinical investigators of the participant centers and of the coordinator
 center and will be analysed and published anonymously and as aggregate, in the respect of the
 current legislation on privacy.
Inclusion Criteria (all of the following):

 -  Children aged 12 months to 12 years

 -  Proven or suspected bacterial infection

 -  Inpatient children needed antibiotic treatment

 -  Oral or intravenous or intramuscular antibiotic administration with at least 2 days of
 duration

 -  Parental informed consent to the participation to the study

 Exclusion Criteria:

 -  Age below 12 months or above 12 years

 -  Infections with no or only one day of antibiotic treatment

 -  Sepsis or severe generalized infection

 -  Pancreatitis

 -  Known immunodeficiency or chronic gastrointestinal diseases or heart disease

 -  Oncologic patients

 -  Severe neurological impairment or lack of verbal expression

 -  Enteral or parenteral nutrition

 -  Gastrointestinal malformations or abdominal surgical interventions

 -  Central line catheter

 -  Acute or chronic diarrhea when starting antibiotics

 -  Current or recent (in the last four weeks) use of any probiotic product

 -  Antibiotic treatment started earlier than 24 hours before enrollment

 -  Absence of parental consent