Young children being breastfed and starting to eat solid foods are at risk of having diarrhea. In our digestive system (in adults but also in infants) live many bacteria that can be good or bad for health. In some cases, the good intestinal bacteria grow poorly and bad bacteria are more numerous. This can cause diarrhea.

With this project, we want to study the effects of adding vitamins and minerals, plus a dietary fiber, called a prebiotic, and a small protein, which is found in breast milk, called lactoferrin, to maize porridge. We want to see if giving these nutrients can improve your child’s nutrition, in particular iron, which is essential for ‘good blood’ and healthy growth and development.

In many countries, flours and cereals already have iron, zinc and vitamins added, and these have been shown to improve health and development in children taking the foods. We need to know if making these additions to food is helpful and whether they cause good or bad intestinal bacteria to grow, and whether they can reduce diarrhea. Therefore, we will be looking at whether feeding infants porridge with these nutrients will improve their health, and also reduce the risk for diarrhea.

The study duration is 9 months, where the participants will be visited weekly and be delivered a micronutrient powder for 6 months, which contains the vitamins, minerals and nutrients, together with 2 kg maize flour for use in the preparation of all porridge fed to the infant. Some infants will receive vitamins and minerals (plus iron), another group will additionally receive the prebiotic, a third group will additionally receive the special lactoferrin protein and a fourth group will additionally receive both the prebiotic and the lactoferrin together. Neither the participants or their caregivers nor the team working in Msambweni will know which infants are in which group. However, a committee of scientists will be receiving health reports on the children and will know which children have received the additional nutrients. If there is any reason to believe that the nutrient powder is affecting the children badly they will stop the study. After the child receives the micronutrient powders for 6 months, he/she will be followed up for 3 months to determine the long-term effects.

The child’s health will be checked at the start of the study and regularly during the study (weekly) and we also want to measure the child’s height/length and weight. At the beginning of the study (Day 1), after 1 month (Day 30), after 6 months (Day 180) (after the child finished taking the micronutrient powder) and at the end of the study (Day 270) (9 months), the child will be asked to give a small sample of blood (the amount in a small spoon) which will be used to see if the child has enough iron in his blood. Enough iron makes a child strong. We will also look at the bacteria in child’s intestine. Caregivers will be therefore asked to collect the child’s stool sample at the beginning (Day 1), after 1 month (Day 30), after 6 months (Day 180) and at the end of the study (Day 270). Caregivers will be shown how these samples have to be collected and stored in a container adding a sachet. These samples will be tested for which bacteria are present. This will help us to observe any changes caused by the nutrient powders.

In some of the infants, we will collect a stool sample prior to study start (Day -1). This sample will be sent to the university in Zurich, Switzerland, where we will run tests on the bacteria.

The ultimate goal of this research is to develop a means to safely administer iron supplements to infants in settings with high infection burden. The hypothesis underlying this project is that promoting development of a beneficial, protective gut microbiota by co-administration of prebiotic galacto-oligosaccharides (GOS) and iron-sequestering bovine lactoferrin (bLF) during iron supplementation will prevent iron-induced increases of opportunistic enteropathogens that cause infection and inflammation. The proposed research will extend our established strategy of conjoining investigations in vivo [1-3]with intestinal fermentation and cellular models in vitro [4-9]. We will conduct a randomized clinical trial in 6-month-old Kenyan infants in conjunction with mechanistic microbiota studies using our established long-term continuous polyfermenter platform inoculated with immobilized fecal microbiota from Kenyan infants.

The period from about 6-12 months of age is vital both for iron nutrition [10-13]and for the establishment of a healthy gut microbiome that promotes immune system development, local immune homeostasis and limits pathogen colonization [14-19]. Oral iron supplements are associated with a significant 15% increase in the rate of diarrhea in children in malaria-endemic areas. Our most recent studies have shown that prebiotic GOS can provide partial amelioration of the adverse effects (AEs) of iron-induced dysbiosis by enhancing the growth of barrier populations of bifidobacteria and lactobacilli [20].  We hypothesize that the combination of prebiotic GOS with bLF has iron sequestration, antimicrobial and immunomodulatory activities [21-26]and will provide virtually complete protection against the AEs of added iron on the intestinal microbiota. This research has two specific aims:

1. To conduct a randomized, controlled double-blind 9-month clinical trial in 6-month old Kenyan infants comparing the effects on gut microbiome composition among groups receiving in-home fortification for 6 months with micronutrient powders (MNPs) containing 5 mg iron (as ferrous fumarate [FeFum]) and (i) GOS (7.5 g), (ii) bLF (1 g), (iii) GOS (7.5 g) and bLF (1 g), and (iv) no GOS or bLF. Each infant will then be followed for an additional 3 months to determine the longer-term effects of the treatments.
2. To examine mechanisms of prebiotic GOS and bLF on microbiota composition, enteropathogen development, microbiota functions and metabolic activity, and inflammatory potential in vitro with treatments paralleling those in Specific Aim 1, using immobilized fecal microbiota from Kenyan infants to inoculate our established long-term continuous polyfermenter intestinal model (PolyFermS) to mimic Kenyan infant colon conditions, together with cellular studies.