Studio Clinico Randomizzato Per Valutare l'Efficacia Del Bifidobacterium BB-12® Nel Trattamento Delle Coliche Infantili
Phase: NA
Conditions: Infantile Colic, Colic, Infantile
Sponsor: SOFAR S.p.A.

 
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PROTOCOL SYNOPSIS 
Study Title 
Randomized Clinical Trial to Evaluate the Efficacy of Bifidobacterium BB-12® in the 
Treatment of Infantile Colic  
Protocol no. 
PSC-DS BIGI 
Sponsor 
SOFAR S.p.A. 
Name of finished 
product 
Bifidobacterium BB-12® 
Investigators and 
study centers 
Prof. Roberto Berni Canani 
Dipartimento di Scienze Mediche Traslazionali – Sezione Pediatrica 
Laboratorio Europeo per lo Studio delle Malattie Indotte da Alimenti 
Università degli Studi di Napoli “Federico II” 
Via S. Pansini 5, Napoli, Italy 
Objectives
The primary objective of this study was to evaluate the proportion of successful treatments, 
defined as a 50% or greater reduction in the average daily duration of crying episodes after 
28 days of treatment.  
The secondary objectives were to evaluate the following variables: 
• 
Decrease from baseline in the number of crying episodes and in the persistence of 
colics at the end of the study (day 28). 
• 
Decrease in the number of regurgitation and vomiting episodes. 
• 
Number of respiratory, gastrointestinal, cutaneous and urinary infections during the 
study. 
• 
Number of bowel movements and consistency of fecal mass. 
Methodology
This was a randomized, double-blind, placebo-controlled clinical study. 
The study was conducted together with five family pediatricians practicing in the city of 
Naples. The pediatricians were asked to refer potentially eligible patients and to provide 
support, if necessary, to their parents. The objectives and procedures of the study were 
thoroughly presented to each pediatrician during a preliminary meeting held at the single 
trial center. During the study, the center was at the disposal of the pediatricians in case of 
need or clarification. 
The Investigator was responsible for enrolling and evaluating the patients, as well as for 
completing the case report forms (CRF).  
After the patient’s parents had provided written informed consent, and once all eligibility 
criteria had been met, patients were required to follow a one-week pre-enrollment period 
during which fermented milk and nutritional supplements containing probiotics were 
prohibited. If after this period the diagnosis of infantile colic was confirmed, the patient 
was randomized in a 1:1 ratio to receive one of the two following products: 
• 
Bifidobacterium animalis subsp. lactis, BB-12®: 6 drops/day (which guaranteed 1 
billion live cells) for 28 consecutive days. 
• 
Placebo oil drops: 6 drops/day for 28 consecutive days. 
The patient’s parents were provided with a diary and were instructed on how to complete 
it with data concerning the daily number and duration of crying episodes, the consistency 
of their baby’s feces (according to the Bristol scale), and the daily number of bowel 
movements. Data was collected from these diaries at each weekly visit. Each patient was 
to undergo a total of 6 visits over a 5-week period.  
Number of patients
The study population was made up of 80 children who met eligibility criteria. 
Diagnosis and main 
criteria for inclusion 
Inclusion criteria 
1. Exclusively breastfed babies of both sexes ≤ 7 weeks of age. 
2. Diagnosis of infantile colic according to Rome III Criteria.

 
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3. Written informed consent of parents.  
Exclusion criteria 
1. Weight at birth < 2500 g. 
2. Gestational age < 37 weeks.  
3. Apgar score at 5 minutes < 7. 
4. Use of baby formulas. 
5. Stunted growth/weight loss (< 100 g/week from birth to last reported weight). 
6. Neurological diseases. 
7. Confirmed or suspected food allergy. 
8. Gastroesophageal reflux disease. 
9. Intake of substances that alter intestinal microbiota (probiotics, prebiotics and 
antibiotics, gastric antacids) in the 2 weeks prior to enrollment. 
10. Fever and/or infectious diseases in the 2 weeks prior to enrollment. 
11. Current systemic infections. 
12. History of congenital infection. 
13. Chronic intestinal disease (cystic fibrosis or other forms of primary pancreatic 
insufficiency). 
14. Primary or secondary gastrointestinal malformations (esophageal atresia, intestinal 
atresia, short bowel syndrome, malrotation). 
15. Metabolic disease. 
16. Genetic diseases and chromosomal abnormalities. 
17. Primary or secondary immunodeficiency. 
18. Insufficient reliability or presence of conditions that make the patient’s compliance 
with the protocol unlikely. 
19. Previous participation in this study. 
Test product, dose and 
mode of 
administration 
• 
Investigational treatment: dietary supplement containing Bifidobacterium (BB-12®) as 
active ingredient. Each 8 ml bottle with dropper contained 1 billion CFU per 6 drops. 
Dosage: 6 drops a day for 28 days. 
OR 
• 
Placebo indistinguishable from the investigational product in packaging, color, weight, 
smell and taste. Dosage: 6 drops a day for 28 days. 
Criteria for evaluation 
Primary efficacy variable 
The primary efficacy variable was the proportion of successful treatments, defined as a 
50% or greater reduction in the average daily duration of crying episodes after 28 days of 
treatment. 
Secondary efficacy variables 
• 
Decrease from baseline in the number of crying episodes and in the persistence of 
colics at the end of the study (day 28) 
• 
Decrease in the number of regurgitation and vomiting episodes 
• 
Number of respiratory, gastrointestinal, cutaneous and urinary infections during the 
study 
• 
Number of bowel movements  
• 
Consistency of fecal mass using the Bristol scale 
• 
Effects of BB-12 on intestinal microbiota 
• 
Variations of the intestinal production of innate immunity peptides (calprotectin, β 
defensin type 2, LL37) and short-chain fatty acids (butyrate e propionate) 
Safety variables 
Safety was assessed based on the results of physical exams and on the incidence of adverse 
events (AE). 
Statistical methods 
The following analysis sets were defined: 
• 
Per Protocol Set (PP): all randomized patients who completed the study without any 
significant protocol violation. 
• 
Intention to Treat Set (ITT): all randomized patients who received at least one dose of 
study treatment. 
• 
Safety Set: all randomized patients who received at least one dose of study treatment. 
Patients were analyzed according to the treatment received. 

 
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A patient who came back for Visit T2 was considered as having received at least one dose 
of study treatment.  
The primary efficacy analysis was performed on the ITT population, and on the PP 
population as supportive. The secondary efficacy analysis was performed on the ITT 
population only. The safety analysis was performed on the Safety population. 
Primary variable 
Mean daily crying duration is described for each week by means of descriptive statistics 
for continuous data and was calculated on non-missing values; observations with values 
equal to zero were included in the computation. Mean change from baseline (mean of Week 
1) was computed for each week. 
The treatment success rate was evaluated in terms of the reduction of crying duration, 
comparing mean daily duration of the last Week (from T4 to T5) with that of Week 1 (from 
T0 to T1). Success was defined as a reduction ≥ 50%. The proportion of patients with 
successful outcomes in the two groups is summarized using descriptive statistics for 
categorical data and analyzed by means of a Chi-Square test. 
Date and version 
English version 1.0, 4 December 2019 – based on Protocol version 3, 16 May 2016 
(document in Italian language)