Effect of Iron Supplements on the Growth of Enteric Pathogens

Study Overview
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Brief Summary
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Iron supplementation is a common approach to address iron deficiency with recommendations for women of childbearing age, particularly those at risk of iron deficiency. Because of its considerably higher absorption, ferrous sulphate is the common iron compound used in iron supplements. However, concerns about iron supplements arise from the knowledge that a large portion of the supplement consumed is not absorbed. This unabsorbed iron travels to the colon and, in preclinical studies, has been shown to promote the growth of enteric pathogens at the expense of beneficial commensal bacteria and increase infection risk, including the clinical incidence of diarrhea. The objective of this study is to compare the effects of iron as ferrous sulfate (FeSO4) or FeSO4-enriched Aspergillus oryzae (Ao iron) on the growth and virulence of common enteric pathogens using an in vitro fecal fermentation model. Stool samples will be collected from women of reproductive age following ingestion of an iron supplement as either FeSO4 or Ao iron. Stool samples will be spiked with common enteric pathogens, and outcome measures will be determined following in vitro fecal fermentation.

Official Title
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In Vitro Fecal Fermentation Following Ingestion of Iron-enriched Aspergillus Oryzae or Iron Sulfate

Conditions
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Iron-deficiency, Iron Deficiency Anaemia Due to Dietary Causes, Iron Deficiency Anemia Treatment, Iron Deficiency Anemia

Intervention / Treatment
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* Dietary Supplement: Ferrous sulfate
* Dietary Supplement: Ferrous sulfate-enriched Aspergillus oryzae (Ao iron)


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Women 18-44 y Are able to swallow the iron pills provided for the study BMI 18.5 to 29.9 kg/m2 Willing to provide blood samples Willing to collect stool samples Exclusion Criteria: Currently taking any antibiotics Iron overload condition/hemochromatosis History of chronic gastrointestinal disorders/diseases Currently smoke cigarettes (including vaping) Have donated blood recently (in the last two weeks) Currently taking a vitamin and mineral supplement containing iron Pregnant and lactating/breastfeeding women Allergic to any of the ingredients (wheat, egg, butter, milk, and baking powder, blue dye) in the muffins

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 44 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Dietary Supplement: FeSO4 Dietary Supplement: Ao iron
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Ferrous sulfate (FeSO4)<br>FeSO4 supplements containing 54 mg elemental iron | Dietary Supplement: Ferrous sulfate<br>* 2 FeSO4 supplements containing 27 mg elemental iron/supplements (54 mg total iron)<br>|
| Experimental: Ferrous sulfate-enriched Aspergillus oryzae (Ao iron)<br>Ao iron supplements containing 54 mg elemental iron | Dietary Supplement: Ferrous sulfate-enriched Aspergillus oryzae (Ao iron)<br>* 2 Ao iron supplements containing 27 mg elemental iron/supplements (54 mg total iron)<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Concentration of iron taken up by enteric pathogens | Iron uptake of common enteric pathogens will be determined in stool following in vitro fecal fermentation. | Time Frame: 0-24 hours | 
| Growth of enteric pathogens measured by optical density | Growth of common enteric pathogens will be determined in stool following in vitro fecal fermentation. | Time Frame: 0-24 hours | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Gut microbiome composition and diversity | Gut microbiome composition will be determined in stool following in vitro fecal fermentation using 16S rRNA gene sequencing. | Time Frame: 0-24 hours | 
| Individual fecal short chain fatty acid (SCFA) concentration | Individual SCFAs will be determined in stool following in vitro fecal fermentation by liquid chromatography-mass spectrometry. | Time Frame: 0-24 hours |