Effectiveness of Fortification With Folic Acid and Vitamin B12 Among Teenage Girls

Does Small Scale Cereal-based Fortification Hold the Key to Improved Micronutrient Status in Ethiopia? The Case of Folic Acid and Vitamin B12 in Teenage Girls in Arba Minch, Ethiopia

Folic acid and vitamin B12 play an interdependent role in key cellular processes, namely deoxyribonucleic acid synthesis, cell division, red blood cell formation, and nervous system myelination. A deficiency of either vitamin will predispose teenagers to many diseases, which persist across their lifespan. Fortification of food with micronutrients has been promoted to reduce micronutrient deficiencies. A large segment of vulnerable populations in low- and middle-income countries (LMICs) resides in rural settings and has limited access to large-scale commercialized fortified foods. In such operational constraints, the use of locally (small-scale) fortified cereals could be an alternative intervention. The study aims to evaluate the effectiveness of small scale folic acid and vitamin B12 fortified cereals in improving folate and vitamin B12 status, growth velocity, puberty status, anaemia, cognitive development and mental health among teenage girls, in rural rift valley of Ethiopia.

Study Overview

• Status: Completed
• Conditions: Depressive Symptoms; Vitamin B 12 Deficiency; Folate Deficiency; Cognitive Development; Anemia, Megaloblastic
• Intervention / Treatment: Other: Unfortified cereal flour; Dietary supplement: Folic acid and vitamin B12 fortified flour

Detailed Description

Folic acid (FA) and vitamin B12 play an interdependent role in key cellular processes, namely deoxyribonucleic acid synthesis, cell division, red blood cell formation, and nervous system myelination. Teenagers are the future mothers. Hence their health and nutritional status have long term consequences on future pregnancies and birth outcomes. Neural Tube Defects (NTDs) are a group of fatal or severely disabling birth defects known to be mainly a consequence of severe folate deficiency in early pregnancy. The policy of advising women to take iron & FA (IFA) supplements during pregnancy has not been successful in preventing NTDs because of the low adherence to antenatal care services including IFA, the large proportion of unplanned pregnancies, the timing at which IFA supplementation starts, and the logistical limitation that makes the access to IFA limited. The evidence is strong on the importance of food fortification in the promotion of maternal and child health mainly through reducing micronutrient deficiencies. A large segment of vulnerable populations in developing countries resides in rural settings and has limited access to fortified foods in the market. In such operational constraints, using locally fortified cereals could be an alternative intervention. However, there is a paucity of evidence regarding effectiveness of small-scale cereal-based fortification; in Ethiopia, the evidence is inexistent. Moreover, teenage girls are an under-studied group and do not constitute a target population from different nutritional intervention programs.

The overall objective of this study is to evaluate the effectiveness of small-scale folic acid and vitamin B12-fortified cereals in improving folate and vitamin B12 status, growth velocity, puberty status, anaemia, cognitive development and mental health among rural teenage girls (13-19 years of age).

This study will be conducted in Arba Minch Health and Demographic Surveillance Sites (AM-HDSS), Southern Ethiopia from October 2023 to April 2024. Effectiveness of intake of folic acid and vitamin B12 fortified meals will be evaluated in a randomized, double-blind controlled trial among 474 teenage girls between 13 and 19 years of age who are residing in and attending one of the schools at AM-HDSS.

Data on the following variables, except for demographic characteristics of teenage girls and their respective parents and/or household heads will be collected at the start and end point of the 6 months intervention. Data collection will be carried out at schools and at respective girls' home.

• Demographic characteristics of the teenage girls, morbidity status, helminthic infections, dietary intake, anthropometry, puberty status, presence of depressive symptoms, biochemical samples and cognitive development will be assessed in teenage girls directly at schools.
• Socio-demographic characteristics of the parents or the head of the household if different from parents, family wealth status, food security, health care services and environmental characteristics will be collected in recruited teenage girls' households within the one week following the enrolment and at the end of the intervention.

• Study Type: Interventional
• Enrollment (Actual): 472
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Ethiopia
  • Arba Minch, Ethiopia
    • Arba Minch University
  • Arba Minch, Ethiopia, 21
    • Arba Minch Health and Demographic Surveillance System sites

Participation Criteria

Eligibility Criteria

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

Description

Inclusion Criteria:

Teenage girls between 13 and 19 years of age who live in households at Arba Minch Health and Demographic Surveillance Sites (AM-HDSS) are eligible to participate in the study if:

• One or both of their parents signed an informed consent form and the girl's agreement.
• Their parents and the girl planned to stay in the study area during the period of the study (minimum 6 months) in the kebele.
• Both parents and girls accept the intervention packages including blood draws and home visits.

Exclusion Criteria:

• Chronically ill girls diagnosed with diabetes and asthma;
• Severely undernourished girls (defined as body mass index z score < -3 standard deviations of the median World Health Organization reference population);
• Severely anaemic girls (Hb concentration <80g/L);
• Teenage girls who are pregnant, lactating or taking IFA/B12 supplements;
• Diagnosed hemoglobinopathy (sickle cell or thalassemia);
• Diagnosed liver diseases like acute hepatitis, cirrhosis, hepatocellular carcinoma, and metastatic liver disease

Study Plan

How is the study designed?

Design Details

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

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Placebo Comparator: Control group; Teenage girls randomly assigned to the control group will receive a standard bag of unfortified cereal flours every week for six months.	Other: Unfortified cereal flour; Standard bags of unfortified cereal-based flours will be prepared at local mills from the common staple cereal food in the area. Participants will be supplied with 6 kg bags of flour, which will be enough for 30 days, and 1.5kg bags of flour will be delivered to their home on a weekly basis and lasting for 6 months. Instructions on the preparation of meals and frequency of intake with a weekly compliance sheet will be provided for monitoring the intervention.; Other Names: Control group
Experimental: folic acid & vit B12 fortified flour; Teenage girls randomly assigned to the control group will receive a standard bag of cereal flours fortified with folic acid and Vit B12 every week for six months.	Dietary supplement: Folic acid and vitamin B12 fortified flour; Standard bags of fortified cereal-based flours will be prepared at local mills from the common staple cereal food in the area. The dose of vitamins will be added to the flour of the intervention group and mixed with batch mixing technique. The concentration of folic acid and vitamin B12 per 1kg of flour will be 2mg and 0.02mg, respectively. Participants will be supplied with 6 kg bags of flour, which will be enough for 30 days, and 1.5kg bags of flour will be delivered to their home on a weekly basis and lasting for 6 months. Instructions on the preparation of meals and frequency of intake with a weekly compliance sheet will be provided for monitoring the intervention.; Other Names: Intervention group

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Serum folic acid concentrations	Serum folate levels ng/ml (nmol/l)	Assessed at 6 months
Erythrocyte (RBC) folic acid concentrations	RBC folate level ng/ml (nmol/l)	Assessed at 6 months
General cognitive ability	Raven Progressive Matrices (RPM) tests, a measure of nonverbal intelligence.	Assessed at 6 months
Working memory	Digit Span (Forward, Backward, and Sequencing) is a standardized test that assesses the working memory of participants.	Assessed at 6 months
Depression	Depressive symptoms will be assessed by Patient Health Questionnaire (PHQ) 9 modified for adolescents (PHQ-9A), a self-report instrument comprised of 9-items. Items are rated on a four-point ordinal scale.	Assessed at 6 months
Serum vitamin B12 concentrations	Serum vitamin B12 level (pmol/L)	Assessed at 6 months
Fat-free mass	An index of adiposity will be measured to evaluate girls' body composition, in %	Assessed at 6 months
Fat mass	An index of adiposity will be measured to evaluate girls' body composition, in kg	Assessed at 6 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
RBC structure (megaloblastic anaemia)	Mean Corpuscular Volume (MCV) in femtoliters/fl	Assessed at 6 months
Plasma homocysteine concentrations	Plasma homocysteine level (µmol/L)	Assessed at 6 months
Weight	Participant weight (kg)	Assessed at 3 and 6 months
Adherence to the intervention/fortification	adherence to the flour consumption will be assessed through home to home visit in a weekly basis.	Assessed weekly for the whole period of intervention (until 6 months)
Haemoglobin concentration	Haemoglobin concentration (g/dl)	Assessed at 6 months
Height	Participant height (cm)	Assessed at 6 months
Puberty status	The pubertal status will be assessed by Pubertal Development Scale which has a 4-point scale ranging from 1 (has not begun) to 4 (development completed). Girls will report on their body hair development, growth spurt, skin changes, breast development and the occurrence of menarche (1=no and 4=yes).	Assessed at 6 months of fortification
Prevalence of soil-transmitted helminths	The presence of worm parasites and egg density in the stools. Three common parasites and their eggs will be investigated, i.e. Ascaris lumbricoides (round worm), Trichuris trichiura (whipworm) and Ancyclostoma duodenale or Necater americanus (hookworms)	Assessed at 6 months
Prevalence of Schistosome infection	The prevalence of Schistosoma mansoni infection	Assessed at 6 months

Collaborators and Investigators

• Sponsor: University Ghent
• Collaborators: Institut de Recherche en Sciences de la Sante, Burkina Faso; Addis Ababa University; Arba Minch University
• Investigators: Principal Investigator: Stefaan De Henauw, MD,PHD, University Ghent

Publications and helpful links

General Publications

• Oumer M, Taye M, Aragie H, Tazebew A. Prevalence of Spina Bifida among Newborns in Africa: A Systematic Review and Meta-Analysis. Scientifica (Cairo). 2020 Oct 6;2020:4273510. doi: 10.1155/2020/4273510. eCollection 2020.
• Wald NJ. Postscript to 'Folic acid and neural tube defects: Discovery, debate and the need for policy change'. J Med Screen. 2022 Sep;29(3):147. doi: 10.1177/09691413221117464. Epub 2022 Aug 8. No abstract available.
• Ardila A. Development of metacognitive and emotional executive functions in children. Appl Neuropsychol Child. 2013;2(2):82-7. doi: 10.1080/21622965.2013.748388. Epub 2013 Jan 28.
• Centeno Tablante E, Pachon H, Guetterman HM, Finkelstein JL. Fortification of wheat and maize flour with folic acid for population health outcomes. Cochrane Database Syst Rev. 2019 Jul 1;7(7):CD012150. doi: 10.1002/14651858.CD012150.pub2.
• Mildon A, Klaas N, O'Leary M, Yiannakis M. Can fortification be implemented in rural African communities where micronutrient deficiencies are greatest? Lessons from projects in Malawi, Tanzania, and Senegal. Food Nutr Bull. 2015 Mar;36(1):3-13. doi: 10.1177/156482651503600101.

Study record dates

Study Major Dates

• Study Start (Actual): September 6, 2023
• Primary Completion (Actual): August 31, 2024
• Study Completion (Actual): August 31, 2024

Study Registration Dates

• First Submitted: October 3, 2023
• First Submitted That Met QC Criteria: October 20, 2023
• First Posted (Actual): October 25, 2023

Study Record Updates

• Last Update Posted (Actual): February 27, 2026
• Last Update Submitted That Met QC Criteria: February 25, 2026
• Last Verified: February 1, 2026

More Information

Terms related to this study

• Keywords: Anemia; Vitamin B12; Folic acid; Cognitive development; Teenage girls; Small scale fortification
• Additional Relevant MeSH Terms: Nutrition Disorders; Behavioral Symptoms; Hematologic Diseases; Avitaminosis; Deficiency Diseases; Malnutrition; Anemia, Macrocytic; Vitamin B Deficiency; Behavior; Nutritional and Metabolic Diseases; Hemic and Lymphatic Diseases; Depression; Anemia; Folic Acid Deficiency; Vitamin B 12 Deficiency; Anemia, Megaloblastic; Heterocyclic Compounds; Heterocyclic Compounds, 2-Ring; Heterocyclic Compounds, Fused-Ring; Investigative Techniques; Epidemiologic Research Design; Epidemiologic Methods; Research Design; Methods; Pterins; Pteridines; Folic Acid; Control Groups
• Other Study ID Numbers: ONZ-2023-0137-C

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

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