Effect of Iron Supplements on the Growth of Enteric Pathogens

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

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.

Study Overview

• Status: Completed
• Conditions: Iron-deficiency; Iron Deficiency Anemia; Iron Deficiency Anemia Treatment; Iron Deficiency Anaemia Due to Dietary Causes
• Intervention / Treatment: Dietary supplement: Ferrous sulfate; Dietary supplement: Ferrous sulfate-enriched Aspergillus oryzae (Ao iron)

• Study Type: Interventional
• Enrollment (Actual): 15
• Phase: Not Applicable

Contacts and Locations

Study Locations

• United States
  • Iowa
    • Ames, Iowa, United States, 50011
      • Iowa State University

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 44 years (Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: Female

Description

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

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Double

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Active Comparator: Ferrous sulfate (FeSO4); FeSO4 supplements containing 54 mg elemental iron	Dietary supplement: Ferrous sulfate; 2 FeSO4 supplements containing 27 mg elemental iron/supplements (54 mg total iron)
Experimental: Ferrous sulfate-enriched Aspergillus oryzae (Ao iron); Ao iron supplements containing 54 mg elemental iron	Dietary supplement: Ferrous sulfate-enriched Aspergillus oryzae (Ao iron); 2 Ao iron supplements containing 27 mg elemental iron/supplements (54 mg total iron)

What is the study measuring?

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

Collaborators and Investigators

• Sponsor: Iowa State University
• Investigators: Principal Investigator: Manju B Reddy, Ph.D., Iowa State University

Study record dates

Study Major Dates

• Study Start (Actual): June 14, 2022
• Primary Completion (Actual): August 2, 2022
• Study Completion (Actual): August 2, 2022

Study Registration Dates

• First Submitted: February 28, 2023
• First Submitted That Met QC Criteria: February 28, 2023
• First Posted (Estimate): March 9, 2023

Study Record Updates

• Last Update Posted (Estimate): March 9, 2023
• Last Update Submitted That Met QC Criteria: February 28, 2023
• Last Verified: September 1, 2022

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Metabolic Diseases; Hematologic Diseases; Nutrition Disorders; Anemia, Hypochromic; Iron Metabolism Disorders; Malnutrition; Anemia, Iron-Deficiency; Anemia; Deficiency Diseases
• Other Study ID Numbers: IVFE

Drug and device information, study documents

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