The Effect of Oral Iron Supplementation on Gut Microbial Composition: a Secondary Analysis of a Double-Blind, Randomized Controlled Trial among Cambodian Women of Reproductive Age
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Study information
-
Quality control
- Retracted paper
- Contamination issues suspected
- Batch effect issues suspected
- Uncontrolled confounding suspected
- Results are suspect (various reasons)
- Tags applied
study design
Citation
PMID PubMed identifier for scientific articles.
DOI Digital object identifier for electronic documents.
URI
Authors
Finlayson-Trick E, Nearing J, Fischer JA, Ma Y, Wang S, Krouen H, Goldfarb DM, Karakochuk CD
Journal
Microbiology spectrum
Year
2023
Keywords:
enteropathogen, gut microbiome, iron supplementation
The World Health Organization recommends untargeted iron supplementation for women of reproductive age (WRA) in countries where anemia prevalence is greater than 40%, such as Cambodia. Iron supplements, however, often have poor bioavailability, so the majority remains unabsorbed in the colon. The gut houses many iron-dependent bacterial enteropathogens; thus, providing iron to individuals may be more harmful than helpful. We examined the effects of two oral iron supplements with differing bioavailability on the gut microbiomes in Cambodian WRA. This study is a secondary analysis of a double-blind, randomized controlled trial of oral iron supplementation in Cambodian WRA. For 12 weeks, participants received ferrous sulfate, ferrous bisglycinate, or placebo. Participants provided stool samples at baseline and 12 weeks. A subset of stool samples (n = 172), representing the three groups, were randomly selected for gut microbial analysis by 16S rRNA gene sequencing and targeted real-time PCR (qPCR). At baseline, 1% of women had iron-deficiency anemia. The most abundant gut phyla were Bacteroidota (45.7%) and Firmicutes (42.1%). Iron supplementation did not alter gut microbial diversity. Ferrous bisglycinate increased the relative abundance of Enterobacteriaceae, and there was a trend towards an increase in the relative abundance of Escherichia-Shigella. qPCR detected an increase in the enteropathogenic Escherichia coli (EPEC) virulence gene, bfpA, in the group that received ferrous sulfate. Thus, iron supplementation did not affect overall gut bacterial diversity in predominantly iron-replete Cambodian WRA, however, evidence does suggest an increase in relative abundance within the broad family Enterobacteriaceae associated with ferrous bisglycinate use. IMPORTANCE To the best of our knowledge, this is the first published study to characterize the effects of oral iron supplementation on the gut microbiomes of Cambodian WRA. Our study found that iron supplementation with ferrous bisglycinate increases the relative abundance of Enterobacteriaceae, which is a family of bacteria that includes many Gram-negative enteric pathogens like Salmonella, Shigella, and Escherichia coli. Using qPCR for additional analysis, we were able to detect genes associated with enteropathogenic E. coli, a type of diarrheagenic E. coli known to be present around the world, including water systems in Cambodia. The current WHO guidelines recommend blanket (untargeted) iron supplementation for Cambodian WRA despite a lack of studies in this population examining iron's effect on the gut microbiome. This study can facilitate future research that may inform evidence-based global practice and policy.
Experiment 1
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Subjects
- Location of subjects
- Cambodia
- Host species Species from which microbiome was sampled. Contact us to have more species added.
- Homo sapiens
- Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology
- Feces Cow dung,Cow pat,Droppings,Dung,Excrement,Excreta,Faeces,Fecal material,Fecal matter,Fewmet,Frass,Guano,Matières fécales@fr,Merde@fr,Ordure,Partie de la merde@fr,Piece of shit,Porción de mierda@es,Portion of dung,Portion of excrement,Portion of faeces,Portion of fecal material,Portion of fecal matter,Portion of feces,Portion of guano,Portion of scat,Portionem cacas,Scat,Spoor,Spraint,Stool,Teil der fäkalien@de,Feces,feces
- Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
- Iron 26Fe,[Fe],Eisen,fer,ferrum,hierro,InChI=1/Fe,InChIKey=XEEYBQQBJWHFJM-UHFFFAOYAG,iron atom,Iron,iron
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Placebo group - 0 week treatment (Baseline)
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- Placebo group - 12 weeks treatment
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Placebo group - 12 weeks treatment refers to patients in the placebo group who are receiving 12 weeks treatment.
- Group 0 sample size Number of subjects in the control (unexposed) group
- 47
- Group 1 sample size Number of subjects in the case (exposed) group
- 47
- Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any)
- 12 weeks
Lab analysis
- Sequencing type
- 16S
- 16S variable region One or more hypervariable region(s) of the bacterial 16S gene
- V6-V8
- Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance
- Illumina
Statistical Analysis
- Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
- relative abundances
- Statistical test
- Beta Binomial Regression
- Significance threshold p-value or FDR threshold used for differential abundance testing (if any)
- 0.05
- MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?
- Yes
Alpha Diversity
- Pielou Quantifies how equal the community is numerically
- unchanged
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Richness Number of species
- unchanged
- Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Source: Figure 4, Figure 5
Description: Differentially abundant bacterial Taxa in the Placebo group.
Abundance in Group 1: increased abundance in Placebo group - 12 weeks treatment
| NCBI | Quality Control | Links |
|---|---|---|
| Enterobacteriaceae | ||
| Enterococcus | ||
| Escherichia/Shigella sp. |
Revision editor(s): Manisha28, KateRasheed
Signature 2
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Source: Figure 4
Description: Differentially abundant bacterial Taxa in the Placebo group.
Abundance in Group 1: decreased abundance in Placebo group - 12 weeks treatment
| NCBI | Quality Control | Links |
|---|---|---|
| Weissella |
Revision editor(s): Manisha28, KateRasheed
Experiment 2
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Ferrous sulfate group - 0 week treatment (Baseline)
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- Ferrous sulfate group - 12 weeks treatment
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Ferrous sulfate group - 12 weeks treatment refers to patients in the Ferrous sulfate group who are receiving 12 weeks treatment.
- Group 0 sample size Number of subjects in the control (unexposed) group
- 40
- Group 1 sample size Number of subjects in the case (exposed) group
- 40
Lab analysis
Statistical Analysis
Alpha Diversity
- Pielou Quantifies how equal the community is numerically
- unchanged
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Richness Number of species
- unchanged
- Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Source: Figure 4
Description: Differentially abundant bacterial Taxa in the Ferrous sulfate group
Abundance in Group 1: increased abundance in Ferrous sulfate group - 12 weeks treatment
| NCBI | Quality Control | Links |
|---|---|---|
| Enterococcus | ||
| Weissella |
Revision editor(s): Manisha28, KateRasheed
Signature 2
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Source: Figure 4, Figure 5
Description: Differentially abundant bacterial Taxa in the Ferrous sulfate group
Abundance in Group 1: decreased abundance in Ferrous sulfate group - 12 weeks treatment
| NCBI | Quality Control | Links |
|---|---|---|
| Enterobacteriaceae | ||
| Escherichia/Shigella sp. |
Revision editor(s): Manisha28, KateRasheed
Experiment 3
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Ferrous bisglycinate group - 0 week treatment (Baseline)
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- Ferrous bisglycinate group - 12 weeks treatment
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Ferrous bisglycinate group - 12 weeks treatment refers to patients in the Ferrous bisglycinate group who are receiving 12 weeks treatment.
- Group 0 sample size Number of subjects in the control (unexposed) group
- 46
- Group 1 sample size Number of subjects in the case (exposed) group
- 46
Lab analysis
Statistical Analysis
Alpha Diversity
- Pielou Quantifies how equal the community is numerically
- unchanged
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Richness Number of species
- unchanged
- Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Source: Figure 4, Figure 5
Description: Differentially abundant bacterial Taxa in the Ferrous bisglycinate group
Abundance in Group 1: increased abundance in Ferrous bisglycinate group - 12 weeks treatment
| NCBI | Quality Control | Links |
|---|---|---|
| Enterobacteriaceae | ||
| Escherichia/Shigella sp. |
Revision editor(s): Manisha28, KateRasheed
Signature 2
Reviewed Marked as Reviewed by Svetlana up on 2025-1-20
Source: Figure 4
Description: Differentially abundant bacterial Taxa in the Ferrous bisglycinate group
Abundance in Group 1: decreased abundance in Ferrous bisglycinate group - 12 weeks treatment
| NCBI | Quality Control | Links |
|---|---|---|
| Enterococcus | ||
| Weissella |
Revision editor(s): Manisha28, KateRasheed
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