Effect of Cow's Milk Kefir on Short Chain Fatty Acid (SCFA), Haemoglobin, and Ferritin Levels of Anemic Adolescent Girls

Effect of Cow's Milk Kefir on Short Chain Fatty Acid (SCFA), Haemoglobin, and Ferritin Levels of Anemic Adolescent Girls in Yogyakarta, Indonesia

Introduction: The prevalence of anemia among adolescent girls in Indonesia is still quite high. According to the Indonesian Health Survey (IHS) in 2023, it was 18% and is still a national problem. Long-term supplementation of blood supplement tablets (TTD) as an effort made by the government to prevent/overcome anemia has caused side effects, namely dysbiosis by pathogenic bacteria. Giving cow's milk kefir containing various probiotics can overcome the side effects of supplementation, and its metabolites accelerate iron absorption so that iron status in the blood increases and ultimately restores anemia in adolescent girls. Method: It's research is an experimental study, with a Randomized Controlled Trial (RCT) design, pre-posttest design, involving research subjects of adolescent girls (15-18 years) with mild and moderate anemia, at SMKN 1, 2, and 3 Kasihan, Bantul Regency, Yogyakarta as many as 80 students. Subjects were divided into 2 groups, namely group 1 (treatment group), which was given 120 ml of cow's milk kefir every day and TTD 1 grain/week, and group 2 (control group), which was given 120 ml of cow's milk every day and TTD 1 grain/week. The duration of administration was 2 months. The study outcomes were Short Chain Fatty Acid (SCFA) levels in feces by Gas Chromatography-Mass Spectrometry (GC-MS) method, hemoglobin by Hematology Analyser method, and serum ferritin by Ferritin ELISA measured at the beginning and end of the study. Statistical analysis included univariate, bivariate, and multivariate analysis. Clinical analysis was also conducted to estimate the effect size of the treatment

Study Overview

• Status: Not yet recruiting
• Conditions: Iron Deficiency Anemia
• Intervention / Treatment: Combination product: cow's milk kefir; Combination product: pasteurized cow's milk

Detailed Description

This protocol represents the only doctoral research dissertation proposal I will undertake as part of my dissertation, which does not exist.

• Study Type: Interventional
• Enrollment (Estimated): 80
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Lusyana Gloria Doloksaribu; Phone Number: 085360637111; Email: lusyanagloriadoloksaribu@mail.ugm.ac.id

Study Locations

• Indonesia
  • Yogyakarta, Indonesia, 55281
    • Gadjah Mada University
    • Contact: Lusyana Gloria Doloksaribu, Doctoral student; Phone Number: 085360637111; Email: lusyanagloriadoloksaribu@mail.ugm.ac.id
    • Contact: Susetyowati, Promotor; Phone Number: 0818277781; Email: susetyowati@ugm.ac.id
    • Principal Investigator: Lusyana Gloria Doloksaribu, Doctoral student
    • Sub-Investigator: Bernadette Josephine Istiti Kandarina, Co-promotor
    • Sub-Investigator: Susetyowati Susetyowati, Promotor

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Adolescent girls aged 15-18 years
• Have a mild and moderate degree of anemia (Hb 8-11.9 g/dl)
• Have a normal menstrual cycle and duration

Exclusion Criteria:

• Suffering from thalassemia, hemosiderosis, blood cancer, and malaria, based on the doctor's diagnosis in the last 3 months
• Taking supplements other than TTD
• Taking antibiotics in the last 3 months
• Suffering from lactose intolerance through interviews

Study Plan

How is the study designed?

Design Details

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

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Active Comparator: intervention group; Fresh cow's milk drink pasteurized at 72 °C, and fermented by kefir grain (5%) for 24 hours at 25 °C, then filtered to produce cow's milk kefir, and stored at 4 °C for 24 hours before drinking, given as 1 bottle/day of 120 ml + 10% liquid sugar for 2 months.	Combination product: cow's milk kefir; group given cow's milk kefir and blood supplement tablets (TTD); Other Names: intervention group
Placebo Comparator: control group; Fresh cow's milk drink pasteurized at 72 °C, without fermentation, given 1 bottle/day of 132 ml for 2 months	Combination product: pasteurized cow's milk; group given pasteurized cow's milk and blood supplement tablets (TTD); Other Names: control group

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Short Chain Fatty Acid	SCFA (acetate, propionate, and butyrate) levels in fresh feces of adolescent girls, using the GC-MS (Gas Chromatography-Mass Spectrometry) Method, were measured before (0 months) and after the intervention (2 months), using a ratio scale. The examination procedure included collecting feces in the field according to the procedure at https://drive.google.com/drive/folders/1uXklwFuC-1rnQ1Anim86b7-9pqLSheIz. Fresh feces samples were collected immediately, by storing them in a cool box (to maintain the quality of feces so that it remains stable during storage and transportation, and avoid the maturation process that can affect the results of lab tests, so that the results of the lab analysis carried out can be accurate and valid), and then immediately taken to the laboratory to quantify each of the short chain fatty acids (SCFA) in the sample.	2 months after intervention
hemoglobin	Adolescent girls' hemoglobin values in the blood taken from the median cubital vein, using the hematology analyzer method (unit g/dl), were measured before (0 months) and after the intervention (2 months), using a ratio scale.	2 months after intervention
Ferritin	Ferritin content in the blood of adolescent girls taken from the median cubital vein, using the Ferritin ELISA method (unit µg/L), was measured before (0 months) and after the intervention (2 months), using a ratio scale	2 months after intervention

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Nutrient intake	The amount of nutrients iron, folic acid, protein, vitamin C, zinc, energy and fat that enter the body of adolescent girls per day obtained from food and beverages, using the Food Record method carried out for 3 days, consisting of 2 times weekday and 1 time weekend, analyzed using the Nutrisurvey 2007 application, measured before (0 months) and after the intervention (2 months), using a ratio scale.	2 months after intervention
Nutritional status of adolescent girls (body mass index / age)	The Z-score or standard deviation (SD) value of adolescent girls after calculating the ratio of body weight in kilograms (kg) squared with height in meters (m) correlated with age, was analyzed using the WHO AnthroPlus application, measured before (0 months) and after the intervention (2 months), using a ratio scale. Weight values were obtained by weighing adolescent girls using digital scales with an accuracy of 0.1 kg. Height values were obtained by measuring the height of adolescent girls using a microtoise with an accuracy of 0.1 cm, and age was obtained by asking about the date/month/year of birth of adolescent girls. Using a questionnaire.	2 months after intervention
Menstrual cycle and duration of adolescent girls	The physiological process experienced by adolescent girls, namely periodic bleeding from the uterus accompanied by the release of the endometrial wall every month, was obtained through a questionnaire, using an ordinal scale, with the categories (1). Normal if the menstrual cycle ranges from 21-35 days, and the length of menstruation is 3-7 days, and (2). Abnormal if the menstrual cycle ranges <21 days and >35 days, and the duration of menstruation is <3 days and >7 days	2 months after intervention
Diarrhea	Complaints of watery or loose bowel movements that occur more than 3 times a day were obtained through a questionnaire, using an ordinal scale, with the categories (1). No diarrhea if the frequency of bowel movements ranged from 1-2 times/day, with a solid consistency, and (2). Diarrhea, the frequency of bowel movements ranging ≥3 times/day, watery, thin consistency	2 months after intervention

Collaborators and Investigators

• Sponsor: Lusyana Gloria Doloksaribu

Study record dates

Study Major Dates

• Study Start (Estimated): May 1, 2025
• Primary Completion (Estimated): December 1, 2025
• Study Completion (Estimated): May 1, 2026

Study Registration Dates

• First Submitted: April 25, 2025
• First Submitted That Met QC Criteria: May 2, 2025
• First Posted (Actual): May 4, 2025

Study Record Updates

• Last Update Posted (Actual): May 4, 2025
• Last Update Submitted That Met QC Criteria: May 2, 2025
• Last Verified: May 1, 2025

More Information

Terms related to this study

• Keywords: Hemoglobin; Iron Deficiency Anemia; Ferritin; Kefir; Short Chain Fatty Acid
• Additional Relevant MeSH Terms: Metabolic Diseases; Hematologic Diseases; Anemia; Iron Metabolism Disorders; Anemia, Hypochromic; Iron Deficiencies; Anemia, Iron-Deficiency
• Other Study ID Numbers: GadjahMada

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: IPD data will be shared, but in the form of coding.
• IPD Sharing Time Frame: IPD can be obtained after the dissertation is registered at the Universitas Gadjah Mada Library or the Faculty of Medicine, Public Health, and Nursing Library
• IPD Sharing Access Criteria: Only investigator
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; ICF; ANALYTIC_CODE; CSR

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No