Faecal microbiota of schoolchildren is associated with nutritional status and markers of inflammation: a double-blinded cluster-randomized controlled trial using multi-micronutrient fortified rice

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Citation
PMID PubMed identifier for scientific articles.
DOI Digital object identifier for electronic documents.
URI
Authors
Seyoum Y, Greffeuille V, Kouadio DKD, Kuong K, Turpin W, M'Rabt R, Chochois V, Fortin S, Perignon M, Fiorentino M, Berger J, Burja K, Ponce MC, Chamnan C, Wieringa FT, Humblot C
Journal
Nature communications
Year
2024
Faecal microbiota plays a critical role in human health, but its relationship with nutritional status among schoolchildren remains under-explored. Here, in a double-blinded cluster-randomized controlled trial on 380 Cambodian schoolchildren, we characterize the impact of six months consumption of two types of rice fortified with different levels of vitamins and minerals on pre-specified outcomes. We investigate the association between the faecal microbiota (16SrRNA sequencing) and age, sex, nutritional status (underweight, stunting), micronutrient status (iron, zinc and vitamin A deficiencies, anaemia, iron deficient anaemia, hemoglobinopathy), inflammation (systemic, gut), and parasitic infection. We show that the faecal microbiota is characterised by a surprisingly high proportion of Lactobacillaceae. We discover that deficiencies in specific micronutrients, such as iron and vitamin A, correlate with particular microbiota profiles, whereas zinc deficiency shows no such association. The nutritional intervention with the two rice treatments impacts both the composition and functions predicted from compositional analysis in different ways. (ClinicalTrials.gov (Identifier: NCT01706419)).

Experiment 1


Needs review

Curated date: 2024/12/12

Curator: Joiejoie

Revision editor(s): Joiejoie

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
Anemia anaemia,anemia,anemia (disease),Anemia
Group 0 name Corresponds to the control (unexposed) group for case-control studies
non anaemic
Group 1 name Corresponds to the case (exposed) group for case-control studies
anaemic
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This Group includes children diagnosed with anemia with hemoglobin levels below the threshold (typically less than 11g/dl).
Group 0 sample size Number of subjects in the control (unexposed) group
304
Group 1 sample size Number of subjects in the case (exposed) group
76
Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any)
30 days

Lab analysis

Sequencing type
16S
16S variable region One or more hypervariable region(s) of the bacterial 16S gene
V3-V4
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
LEfSe
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)?
No
LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool
2.0
Confounders controlled for Confounding factors that have been accounted for by stratification or model adjustment
age, sex

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/16

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 A

Description: Differential ASVs between the children grouped according to their nutritional status. anaemia.

Abundance in Group 1: increased abundance in anaemic

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ErysipelatoclostridiaceaeErysipelatoclostridiaceae

Revision editor(s): Joiejoie

Signature 2

Needs review

Curated date: 2024/12/16

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 A

Description: Differential ASVs between the children grouped according to their nutritional status. anaemia.

Abundance in Group 1: decreased abundance in anaemic

NCBI Quality ControlLinks
Prevotella_7Prevotella_7

Revision editor(s): Joiejoie

Experiment 2


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Iron deficiency anemia Ferropenic,Iron deficiency anaemia,Iron-deficiency anaemia,Iron-deficiency anemia,Iron deficiency anemia,iron deficiency anemia
Group 0 name Corresponds to the control (unexposed) group for case-control studies
no iron deficiency anaemia
Group 1 name Corresponds to the case (exposed) group for case-control studies
iron deficient anaemia
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children diagnosed with iron deficient anemia with ferritin levels below 12 µg/L in children
Group 0 sample size Number of subjects in the control (unexposed) group
333
Group 1 sample size Number of subjects in the case (exposed) group
47

Lab analysis

Statistical Analysis

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/16

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 B

Description: Differential ASVs between the children grouped according to their nutritional status. Iron deficiency anaemia.

Abundance in Group 1: increased abundance in iron deficient anaemia

NCBI Quality ControlLinks
PrevotellaceaeNK3831groupPrevotellaceaeNK3831group

Revision editor(s): Joiejoie

Signature 2

Needs review

Curated date: 2024/12/16

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 B

Description: Differential ASVs between the children grouped according to their nutritional status. Iron deficiency anaemia.

Abundance in Group 1: decreased abundance in iron deficient anaemia

NCBI Quality ControlLinks
Provotella_7Provotella_7

Revision editor(s): Joiejoie

Experiment 3


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Vitamin A deficiency deficiencies, vitamin A,deficiency, vitamin A,vitamin A deficiencies,Vitamin A Deficiency,vitamin A deficiency,vitamin A deficiency (disease),Vitamin A deficiency
Group 0 name Corresponds to the control (unexposed) group for case-control studies
Sufficient vitamin A
Group 1 name Corresponds to the case (exposed) group for case-control studies
Vitamin A deficiency
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children diagnosed with Vitamin A deficiency with level less than 20 micrograms per deciliter [mcg/dL]
Group 0 sample size Number of subjects in the control (unexposed) group
350
Group 1 sample size Number of subjects in the case (exposed) group
30

Lab analysis

Statistical Analysis

MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?
Yes

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/16

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 C

Description: Differential ASVs between the children grouped according to their nutritional status. Vitamin A deficiency.

Abundance in Group 1: increased abundance in Vitamin A deficiency

NCBI Quality ControlLinks
Desulfovibrionaceae

Revision editor(s): Joiejoie

Signature 2

Needs review

Curated date: 2024/12/16

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 C

Description: Differential ASVs between the children grouped according to their nutritional status. Vitamin A deficiency

Abundance in Group 1: decreased abundance in Vitamin A deficiency

NCBI Quality ControlLinks
Gordonibacter

Revision editor(s): Joiejoie

Experiment 4


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Growth delay Delayed growth,Growth deficiency,Growth delay,Growth failure,Growth retardation,Poor growth,Retarded growth,Very poor growth,growth delay
Group 0 name Corresponds to the control (unexposed) group for case-control studies
non-stunted children
Group 1 name Corresponds to the case (exposed) group for case-control studies
stunted children
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children diagnosed with stunting with anthropometry measurement showing its prevalence at 40.0% at baseline
Group 0 sample size Number of subjects in the control (unexposed) group
208
Group 1 sample size Number of subjects in the case (exposed) group
172

Lab analysis

Statistical Analysis

MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?
No

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/17

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 D

Description: Differential ASVs between the children grouped according to Stunting

Abundance in Group 1: increased abundance in stunted children

NCBI Quality ControlLinks
Alistipes

Revision editor(s): Joiejoie

Signature 2

Needs review

Curated date: 2024/12/17

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 2 D

Description: Differential ASVs between the children grouped according to stunting

Abundance in Group 1: decreased abundance in stunted children

NCBI Quality ControlLinks
Prevotella_7Prevotella_7
Holdemanella

Revision editor(s): Joiejoie

Experiment 5


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Nutritional deficiency disease malnourished,malnutrition,nutritional deficiency disease,Nutritional deficiency disease
Group 0 name Corresponds to the control (unexposed) group for case-control studies
UR-original
Group 1 name Corresponds to the case (exposed) group for case-control studies
UR-Improved
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children placed on the more diverse micronutrient-fortified rice (FORISCA) fortified with vitamins A, B3, and B12 as additional vitamins.
Group 0 sample size Number of subjects in the control (unexposed) group
114
Group 1 sample size Number of subjects in the case (exposed) group
122

Lab analysis

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
arcsine square-root
Statistical test
MaAsLin2

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/17

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 4

Description: Impact of the nutritional intervention on the taxonomic composition of the faecal microbiota of the 380 children

Abundance in Group 1: increased abundance in UR-Improved

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Agathobacter

Revision editor(s): Joiejoie

Signature 2

Needs review

Curated date: 2024/12/17

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Figure 4

Description: Impact of the nutritional intervention on the taxonomic composition of the faecal microbiota of the 380 children

Abundance in Group 1: decreased abundance in UR-Improved

NCBI Quality ControlLinks
CAG.56CAG.56

Revision editor(s): Joiejoie

Experiment 6


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Inflammation inflammatory response,Inflammation,inflammation
Group 0 name Corresponds to the control (unexposed) group for case-control studies
No inflammation
Group 1 name Corresponds to the case (exposed) group for case-control studies
Inflammation
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children grouped according to systemic inflammation
Group 0 sample size Number of subjects in the control (unexposed) group
240
Group 1 sample size Number of subjects in the case (exposed) group
140

Lab analysis

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
relative abundances
Statistical test
LEfSe

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/17

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Supplementary Figure 2:

Description: Differential ASVs between the children grouped according to their systemic inflammation.

Abundance in Group 1: increased abundance in Inflammation

NCBI Quality ControlLinks
Prevotella

Revision editor(s): Joiejoie

Signature 2

Needs review

Curated date: 2024/12/17

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Supplementary Figure 2:

Description: Differential ASVs between the children grouped according to A) their systemic inflammation.

Abundance in Group 1: decreased abundance in Inflammation

NCBI Quality ControlLinks
Agathobacter

Revision editor(s): Joiejoie

Experiment 7


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Parasitic infection animal parasitic infectious disease,central nervous system parasitic infectious disease,Disease caused by parasite,disease caused by parasite,Disease caused by parasite (disorder),Disease caused by parasite, NOS,Disease, Parasitic,disease, parasitic,Diseases, Parasitic,diseases, parasitic,Ectoparasitic disease,ectoparasitic disease,Infestation,infestation,Infestation, NOS,intestinal parasitism (disorder),intestinal parasitism NOS (disorder),intestinal parasitism, unspecified,intestinal parasitism, unspecified (disorder),Parasite infestation,parasite infestation,Parasite infestation, NOS,parasitemia,parasitic animal,PARASITIC DIS,Parasitic disease,parasitic disease,Parasitic disease, NOS,Parasitic Diseases,parasitic endophthalmitis,Parasitic endophthalmitis (disorder),Parasitic endophthalmitis NOS,Parasitic endophthalmitis NOS (disorder),parasitic eye infection,Parasitic eye infection (disorder),parasitic eye infectious disease,parasitic gastrointestinal disorder,parasitic infection,Parasitic infection of lung,Parasitic infection of skin,Parasitic infection, NOS,parasitic infectious disease,Parasitic infectious disease of the central nervous system (disorder),Parasitic infectious disease, NOS,parasitic infestation of orbit,Parasitic infestation of orbit (disorder),parasitic intestinal infectious disease,parasitic liver disorder,parasitic liver infectious disease,parasitic lung infectious disease,parasitic pregnancy complication,parasitic skin disorder,parasitic skin infectious disease,Parasitic skin infestation (disorder),Parasitism,parasitism,Parasitism, NOS,Parasitosis,parasitosis,Parasitosis, NOS,psorospermiasis,Psorospermiasis (disorder),skin Infestation NOS,Parasitic infection
Group 0 name Corresponds to the control (unexposed) group for case-control studies
No parasitic infection
Group 1 name Corresponds to the case (exposed) group for case-control studies
Parasitic infection
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children grouped according to parasitic infection
Group 0 sample size Number of subjects in the control (unexposed) group
276
Group 1 sample size Number of subjects in the case (exposed) group
104

Lab analysis

Statistical Analysis

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Experiment 8


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Age Age,age
Group 0 name Corresponds to the control (unexposed) group for case-control studies
10-14 years old
Group 1 name Corresponds to the case (exposed) group for case-control studies
6-9 years old
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children grouped according to age (6-9 years old)
Group 0 sample size Number of subjects in the control (unexposed) group
168
Group 1 sample size Number of subjects in the case (exposed) group
212

Lab analysis

Statistical Analysis

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased

Signature 1

Needs review

Curated date: 2024/12/18

Curator: Joiejoie

Revision editor(s): Joiejoie

Source: Supplementary Figure 2:

Description: Differential ASVs between the children grouped according to age

Abundance in Group 1: increased abundance in 6-9 years old

NCBI Quality ControlLinks
Prevotella

Revision editor(s): Joiejoie

Experiment 9


Needs review

Curated date: 2024/12/15

Curator: Joiejoie

Revision editor(s): Joiejoie

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Biological sex Biological sex,biological sex
Group 0 name Corresponds to the control (unexposed) group for case-control studies
Girl
Group 1 name Corresponds to the case (exposed) group for case-control studies
Boy
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
This group includes children identified on sex as a covariate feature.
Group 0 sample size Number of subjects in the control (unexposed) group
180
Group 1 sample size Number of subjects in the case (exposed) group
200

Lab analysis

Statistical Analysis

Alpha Diversity

Pielou Quantifies how equal the community is numerically
decreased
Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample
decreased