New insights into the combined effects of aflatoxin B1 and Eimeria ovinoidalis on uterine function by disrupting the gut-blood-reproductive axis in sheep
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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
Huang SC, Liu KL, Chen P, Xu BW, Ding WL, Yue TJ, Lu YN, Li SY, Li JK, Jian FC
Journal
Microbiome
Year
2024
Keywords:
E. ovinoidalis, Aflatoxin B1, Coccidia, Gut microbiota, Mycotoxin, Reproduction toxicity
BACKGROUND: Sheep coccidiosis is an infectious parasitic disease that primarily causes diarrhea and growth retardation in young animals, significantly hindering the development of the sheep breeding industry. Cereal grains and animal feeds are frequently contaminated with mycotoxins worldwide, with aflatoxin B1 (AFB1) being the most common form. AFB1 poses a serious threat to gastrointestinal health upon ingestion and affects the function of parenteral organs, thus endangering livestock health. However, the impact of the combined effects of coccidia and AFB1 on the reproductive system of sheep has not been reported. Therefore, this study utilized sheep as an animal model to investigate the mechanisms underlying the reproductive toxicity induced by the individual or combined effects of AFB1 and Eimeria ovinoidalis (E. ovinoidalis) on the gut-blood-reproductive axis. RESULTS: The results showed that AFB1 and coccidia adversely affect the reproductive system defense of sheep by altering uterine histopathology and hormone levels and triggering inflammation, which is associated with changes in the gut microbiota and metabolites. Moreover, co-exposure to AFB1 and coccidia disrupted the intestinal structure of the colon, resulting in reduced crypt depth. The impaired barrier function of the colon manifests primarily through the suppression of barrier protein expression, changes in the gut microbiome composition, and disruptions in gut metabolism. Importantly, the levels of blood inflammatory factors (IL-6, IL-10, TNF-α, and LPS) increased, suggesting that exposure to AFB1 and coccidia compromises the function of uterine organs in sheep by perturbing the gut-blood-reproductive axis. Blood metabolomics analysis further revealed that the differential metabolites predominantly concentrate in the amino acid pathway, particularly N-acetyl-L-phenylalanine. This metabolite is significantly correlated with IL-6, TNF-α, LPS, ERα, and ERβ, and it influences hormone levels while inducing uterine damage through the regulation of the downstream genes PI3K, AKT, and eNOS in the relaxin signaling pathway, as demonstrated by RNA sequencing. CONCLUSIONS: These findings reveal for the first time that the combined effects of AFB1 and E. ovinoidalis on sheep uterine function operate at the level of the gut-blood-reproductive axis. This suggests that regulating gut microbiota and its metabolites may represent a potential therapeutic strategy for addressing mycotoxins and coccidia-co-induced female reproductive toxicity.
Experiment 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Subjects
- Location of subjects
- China
- Host species Species from which microbiome was sampled. Contact us to have more species added.
- Ovis aries
- 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
- Treatment outcome measurement Treatment outcome measurement,treatment outcome measurement
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Combination of Con(control), AFB1 (Aflatoxin B1), AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment groups
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- E.0 (Eimeria ovinoidalis) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- A treatment group with E.0 (Eimeria ovinoidalis) which is a coccidian parasite that has been identified as the most pathogenic species among sheep worldwide and received a single oral administration of 2 × 104 sporulated oocysts on day 0 only.
- Group 0 sample size Number of subjects in the control (unexposed) group
- 12
- Group 1 sample size Number of subjects in the case (exposed) group
- 4
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
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3f
Description: Linear discriminant analysis effect size (LEfSe) comparison analysis between the groups.
Abundance in Group 1: increased abundance in E.0 (Eimeria ovinoidalis) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Bacteroidales | ||
| Bacteroidia | ||
| Bacteroidota | ||
| Pseudomonadota | ||
| Gammaproteobacteria |
Revision editor(s): Tosin
Experiment 2
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Combination of Con(control), E.0 (Eimeria ovinoidalis), AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment groups
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- AFB1 (Aflatoxin B1) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Treatment group with Aflatoxin B1 which is a mycotoxin produced by Aspergillus flavus and Aspergillus parasiticus, is a member of the aflatoxin family and is commonly found in cereals and animal feed all over the world, together with other fungal toxins and received daily oral administration of AFB1 at a dosage of 60 μg/kg
Lab analysis
Statistical Analysis
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3f
Description: Linear discriminant analysis effect size (LEfSe) comparison analysis between the groups.
Abundance in Group 1: increased abundance in AFB1 (Aflatoxin B1) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Clostridia | ||
| Eubacteriales | ||
| Oscillospiraceae | ||
| Verrucomicrobiales | ||
| Verrucomicrobiia | ||
| Verrucomicrobiota | ||
| Akkermansia | ||
| Verrucomicrobiaceae |
Revision editor(s): Tosin
Experiment 3
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Combination of Con(control), E.0 (Eimeria ovinoidalis) and AFB1 (Aflatoxin B1) treatment groups
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Treatement group that received a single oral administration of 2 × 104 sporulated oocysts on day 0 along with daily oral administration of AFB1 at a dosage of 60 μg/kg.
Lab analysis
Statistical Analysis
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3f
Description: Linear discriminant analysis effect size (LEfSe) comparison analysis between the groups
Abundance in Group 1: increased abundance in AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Lachnospiraceae | ||
| Clostridiaceae | ||
| Clostridium | ||
| Dorea | ||
| Bacteroides | ||
| Bacteroidaceae |
Revision editor(s): Tosin
Experiment 4
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Combination of E.0 (Eimeria ovinoidalis), AFB1 (Aflatoxin B1) and AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis treatment groups
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- Control (Con) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Treatment group that received daily oral administration of PBS solution and DMSO.
Lab analysis
Statistical Analysis
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3f
Description: Linear discriminant analysis effect size (LEfSe) comparison analysis between the groups
Abundance in Group 1: increased abundance in Control (Con) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Bacillota | ||
| Erysipelotrichaceae | ||
| Erysipelotrichales | ||
| Erysipelotrichia | ||
| Muribaculaceae | ||
| p_75_a5p_75_a5 | ||
| Eubacterium |
Revision editor(s): Tosin
Experiment 5
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- AFB1 (Aflatoxin B1) treatment group
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Treatment group that received a single oral administration of 2 × 104 sporulated oocysts on day 0 along with daily oral administration of AFB1 at a dosage of 60 μg/kg
- Group 0 sample size Number of subjects in the control (unexposed) group
- 4
Lab analysis
Statistical Analysis
- Statistical test
- Dunn's test
- LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool
- Not specified
Alpha Diversity
- Shannon Estimator of species richness and species evenness: more weight on species richness
- decreased
- Chao1 Abundance-based estimator of species richness
- decreased
- Simpson Estimator of species richness and species evenness: more weight on species evenness
- decreased
- Richness Number of species
- decreased
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3i
Description: Relative abundance analysis of common differential genera
Abundance in Group 1: increased abundance in AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Bacteroides | ||
| Blautia | ||
| Candidatus Epulonipiscium |
Revision editor(s): Tosin
Experiment 6
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- E.0 (Eimeria ovinoidalis) treatment group
Lab analysis
Statistical Analysis
Alpha Diversity
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Chao1 Abundance-based estimator of species richness
- unchanged
- Simpson Estimator of species richness and species evenness: more weight on species evenness
- unchanged
- Richness Number of species
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3E and 3I
Description: Relative abundance analysis of common differential genera
Abundance in Group 1: decreased abundance in AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Bacteroidota | ||
| Desulfovibrio |
Revision editor(s): Tosin
Experiment 7
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Con (Control) treatment group
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- AFB1 (Aflatoxin B1) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Treatment group with Aflatoxin B1 which is a mycotoxin produced by Aspergillus flavus and Aspergillus parasiticus, is a member of the aflatoxin family and is commonly found in cereals and animal feed all over the world, together with other fungal toxins and received daily oral administration of AFB1 at a dosage of 60 μg/kg
Lab analysis
Statistical Analysis
Alpha Diversity
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Chao1 Abundance-based estimator of species richness
- unchanged
- Simpson Estimator of species richness and species evenness: more weight on species evenness
- unchanged
- Richness Number of species
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3I
Description: Relative abundance analysis of common differential genera
Abundance in Group 1: decreased abundance in AFB1 (Aflatoxin B1) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Eubacterium |
Revision editor(s): Tosin
Experiment 8
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- E.0 (Eimeria ovinoidalis) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- A treatment group with E.0 (Eimeria ovinoidalis) which is a coccidian parasite that has been identified as the most pathogenic species among sheep worldwide and received a single oral administration of 2 × 104 sporulated oocysts on day 0 only.
Lab analysis
Statistical Analysis
Alpha Diversity
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Chao1 Abundance-based estimator of species richness
- unchanged
- Simpson Estimator of species richness and species evenness: more weight on species evenness
- increased
- Richness Number of species
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3E and 3I
Description: Relative abundance analysis of common differential genera
Abundance in Group 1: decreased abundance in E.0 (Eimeria ovinoidalis) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Bacillota | ||
| Eubacterium |
Revision editor(s): Tosin
Experiment 9
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Differences from previous experiment shown
Subjects
- Group 0 name Corresponds to the control (unexposed) group for case-control studies
- Con(control) treatment group
- Group 1 name Corresponds to the case (exposed) group for case-control studies
- AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
- Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
- Treatement group that received a single oral administration of 2 × 104 sporulated oocysts on day 0 along with daily oral administration of AFB1 at a dosage of 60 μg/kg.
Lab analysis
Statistical Analysis
Alpha Diversity
- Shannon Estimator of species richness and species evenness: more weight on species richness
- unchanged
- Chao1 Abundance-based estimator of species richness
- unchanged
- Simpson Estimator of species richness and species evenness: more weight on species evenness
- unchanged
- Richness Number of species
- unchanged
Signature 1
Reviewed Marked as Reviewed by Svetlana up on 2025-3-28
Source: Figure 3I
Description: Relative abundance analysis of common differential genera
Abundance in Group 1: decreased abundance in AFB1 + E0 (Aflatoxin B1 + Eimeria ovinoidalis) treatment group
| NCBI | Quality Control | Links |
|---|---|---|
| Campylobacter | ||
| Succinivibrio | ||
| Candidatus Epulonipiscium |
Revision editor(s): Tosin
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