Multi-Nutrient Fortified Dairy-Based Drink Reduces Anaemia without Observed Adverse Effects on Gut Microbiota in Anaemic Malnourished Nigerian Toddlers: A Randomised Dose-Response Study

Abstract

Prevalence of anaemia among Nigerian toddlers is reported to be high, and may cause significant morbidity, affects brain development and function, and results in weakness and fatigue. Although, iron fortification can reduce anaemia, yet the effect on gut microbiota is unclear. This open-label randomised study in anaemic malnourished Nigerian toddlers aimed to decrease anaemia without affecting pathogenic gut bacteria using a multi-nutrient fortified dairy-based drink. The test product was provided daily in different amounts (200, 400 or 600 mL, supplying 2.24, 4.48 and 6.72 mg of elemental iron, respectively) for 6 months. Haemoglobin, ferritin, and C-reactive protein concentrations were measured to determine anaemia, iron deficiency (ID) and iron deficiency anaemia (IDA) prevalence. Faecal samples were collected to analyse gut microbiota composition. All three dosages reduced anaemia prevalence, to 47%, 27% and 18%, respectively. ID and IDA prevalence was low and did not significantly decrease over time. Regarding gut microbiota, Enterobacteriaceae decreased over time without differences between groups, whereas Bifidobacteriaceae and pathogenic E. coli were not affected. In conclusion, the multi-nutrient fortified dairy-based drink reduced anaemia in a dose-dependent way, without stimulating intestinal potential pathogenic bacteria, and thus appears to be safe and effective in treating anaemia in Nigerian toddlers.

Keywords: Nigeria; anaemia; iron deficiency; iron deficiency anaemia; malnourished; microbiota; multi-nutrient fortified dairy-based drink; toddler.

Figure 1

Flow-chart of screening and randomisation process.

Figure 2

RDA on the operational taxonomic unit (OTU) level, assessing the effect of time (including test formula treatment) on gut microbiota composition. The covariance attributable to the subject was first fitted by regression and then partially out (removed) from the ordination. OTUs were used as response data and the time point was explanatory data, the bacterial families that contributed most were plotted supplementary. Variation explained by time point was 16.8%, p = 0.002.

Figure 3

Effect of daily intakes of 200, 400, or 600 mL of YCF during 6 mo. on Enterobacteriaceae, pathogenic E. coli and Bifidobacteriaceae. Boxplots are displayed as Tukey whiskers. (A) Relative abundance of Enterobacteriaceae at baseline and endline. At endline relative abundance was significantly higher in the 400 mL group compared to the 600 mL group (p < 0.05 based on Dunn’s posthoc test). (B) Change in the relative abundance of Enterobacteriaceae over time (2log ratio of relative abundance at endline and baseline). (C) Abundance of pathogenic E. coli at baseline and endline (pathogenic E. coli represents the sum of the log gene copies of EPEC, ETEC lt and ETEC st). (D) Change in abundance of pathogenic E. coli over time (delta gene copies). (E) Relative abundance of Bifidobacteriaceae at baseline and endline. (F) Change in the relative abundance of Bifidobacteriaceae over time (2log ratio of relative abundance at endline and baseline).