Effects of a food enriched with probiotics based Lactobacillus rhamnosus GG on the count of Streptococcus mutans and Lactobacillus sp and the presence of incipient caries in children 3 to 4 years in Cali, Colombia

• Register: ANZCTR
• Last refreshed on: 13 January 2020
• Main ID: ACTRN12616001363471
• Date of registration: 30/09/2016
• Prospective Registration: Yes
• Primary sponsor: Universidad del Valle
• Public title: Effects of a food enriched with probiotics based Lactobacillus rhamnosus GG on the count of Streptococcus mutans and Lactobacillus sp and the presence of incipient caries in children 3 to 4 years in Cali, Colombia
• Scientific title: Effects of a food enriched with probiotics based Lactobacillus rhamnosus GG on the count of Streptococcus mutans and Lactobacillus sp and the presence of incipient caries in children 3 to 4 years in Cali, Colombia
• Date of first enrolment: 15/10/2016
• Target sample size: 360
• Recruitment status: Completed
• URL: https://anzctr.org.au/ACTRN12616001363471.aspx
• Study type: Interventional
• Study design: Purpose: Prevention; Allocation: Randomised controlled trial; Masking: Blinded (masking used);Assignment: Parallel;Type of endpoint: Efficacy;
• Phase: Phase 3

Countries of recruitment

Colombia

Contacts

• Name: Prof Judy Villavicencio

• Name: Prof Judy Villavicencio

Key inclusion & exclusion criteria

Inclusion criteria: Healthy children from a general health perspective, with or without cavitated carious lesions at during baseline and tracing three, six, nine and twelve months with ages of 3 years to 4 years, both genders, of the Colombian Institute for family welfare (ICBF) belonging at low socio-economic stratum in the city of Cali, Colombia.
Exclusion criteria: Children with systemic disorders who need special care and / or who have intolerance to milk drinks or allergy to any of the components of the experimental and / or placebo beverage and children who are under permanent antibiotic treatment

Age minimum: 3 Years
Age maximum: 4 Years
Gender: Both males and females

Health Condition(s) or Problem(s) studied

Oral and Gastrointestinal - Other diseases of the mouth, teeth, oesophagus, digestive system including liver and colon

Infection - Studies of infection and infectious agents

Dental caries ;
Dental caries

Intervention(s)

A total number of 360 children were cluster randomized assigned to two groups: children in the intervention group were served milk supplemented with probiotic and the control group received standard milk.

The children will be examined at baseline, the pH in saliva will be taken, samples of saliva, plaque index. Their teeth will be brush and them make the evaluation of dental caries with the International Caries Detection and Assessment System (ICDAS).

It will start the supply of milk with probiotics and without probiotics. At three, six, nine and twelve months milk consumption controls will be made to children.

Controls consist of taking the pH before the milk consumption. Pass an hour it will take again to the children salivary pH; saliva samples, the plaque index. The teeth will be brush and them make the evaluation of dental caries with the International Caries Detection and Assessment System (ICDAS).

The milk supplemented with probiotic contain two probiotic strains (Lactobacillus rhamnosus 5x106 UFC/g and Bifidobacterium Longum 3x106 UFC/g) "Registered Trademark". The milk (test and placebo), identical in taste and appearance, will be supplied in packages separated by the colors "White" or "Blue",

The chefs prepare 200 grams of milk to the children every day for five days a week, for twelve months and are served at breakfast. It is made daily children attendance record and the consumption of milk.

Parents will be instructed about the report to the staff of the project's adverse effects on the children immediately happen.

Primary Outcome(s)

Count of colony-forming units' (CFU) of Streptococcus mutans and Lactobacillus sp.

Participants will be requested to void saliva into a wide-mouthed disposable tube after their breakfast. Saliva (0,1 mL) will be diluted with 0,9 mL of sterile saline solution and tenfold serial dilutions of saliva will be made up to 103, before plating.

Mitis Salivarius Agar (MSA) supplemented with potassium tellurite and bacitracine will be used as culture media for isolation of total salivary Streptococci and Rogosa agar will be prepared for the growth of Lactobacilli. Appropriate amounts of semisolid media will be poured onto petri plates in a laminar air flow unit and allowed to solidify at room temperature.

To each plate 100 microlitres of the diluted sample will be spread evenly and the plates will be sealed, placed in an anaerobic jar with Anaero Gas Pack system, and incubated at 37 degrees Celsius for 48 hours. Bacterial colonies will be observed under a dissecting microscope and deduced on the basis of colony morphology, shape and color; and in case of doubt confirmation for Streptococci and Lactobacilli will be done by gram staining or catalase tests, respectively. The number of CFU will be multiplied by the number of times the original mL of sample will be diluted (the dilution factor of the plate counted) and expressed as the number of colony forming units per milliliter (CFU/mL) of saliva.
[12 months]

Secondary Outcome(s)

3. Salivary pH: For pH saliva, the testing lead tape into mouth saliva for three seconds, is removed and compare to color chart of pH testing tape it to get a value of pH, digital pH meter analyzed the collected saliva, during baseline and tracing at three, six, nine and twelve months
[Baseline 3, 6, 9 and 12 months]

4.Salivary buffer capacity: Measured salivary pH values both before and after addition of acid (HCl) to assess salivary buffer capacity, It was obtained by mixing 1 ml of saliva with 3 ml HCl in a tube and measuring the solution pH with digital pH meter after 20 minutes in vortex mixer, during baseline and tracing at three, six, nine and twelve months[baseline 3, 6, 9 and 12 months]

1. Dental caries lesions: Clinical dental examinations will be performed at baseline and tracing at three, six, nine and twelve months of the study. One dentist, using an artificial light, a mouth mirror, will examine all of the children at the nursery schools. ICDAS (International Caries Detection and Assessment System) criteria were used for the visual and tactile detection of dental caries lesions and for describing the severity of lesions. The essential elements of the ICDAS caries detection codes, are the visual criteria for the full series of six caries codes, using from the histologically more extensive distinct visual change which is visible wet or dry from code 2 through steps to the most severe stage, the extensive distinct cavity with visible dentine until code 6, during baseline and tracing three, six, nine and twelve months. [baseline, 3, 6, 9 and 12 months. ]

2. Presence of bacterial plaque: Using a mouth mirror, gauze, and a dental probe under a focusable flashlight was recorded the presence or absence of clinically visible plaque, during baseline and tracing three, six, nine and twelve months

[Baseline, 3, 6 , 9 and 12 months]

Secondary ID(s)

none

Source(s) of Monetary Support

Colciencias

Ethics review

Status: Approved
Approval date:
Contact:

Universidad del Valle