Gut microbiome composition: link between sports performance and protein absorption?
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g
and Csilla Oláhh
1. Introduction
A sufficient protein intake is essential for an adequate physical condition and athletic performance.Nowadays the study of the composition and the role of the gut microbiome in relation to sport nutrition have become an increasingly researched area.
This study aimed to examined how short-term diet of plant-based vegan protein affected muscle development in elite water polo players, in a 31-day period. It also investigated association between physical condition of the athletes and the composition of their gut microbiome after protein supplement with or without pre- and probiotics.(During the experiment none of the players were under any medical care, got any antibiotics or other treatments that could have influenced the microbiome composition)
INT group
CON group
(n=10)
(n=10)
- Vegan protein supplement
- Prebiotics
- Fermented probiotic herbal product
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
2. Study design
INT group
CON group
They receive: 250 ml of Biotech vegan protein shake daily during the 31-day study
They receive: 250 ml of Biotech vegan protein shake daily during the 31-day study 15 drops of Herbaferm Probiotic solution, dissolved in 2 dl of water immediately after waking up 3.5 g of prebiotic mixture, homogenised daily in their 250 ml vegan protein shake
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
3. Analysis and data collection
Anthropometric and body composition analysis was performed by InBody 970, and several parameters were determined, such as weight, body fat mass, fat-free mass, skeletal muscle mass, percent body fat, visceral fat area, body cell mass, arm circumference, arm muscle circumference, protein mass, basal metabolic rate and overall fitness score (InBody Score).
Also blood samples were collected, and the parameter analysed were the count of red blood cells, value of haemoglobin, hematocrit, MCV, MCH, MCHC and vitamin B12 level.
The Bristol stool scale chart was used. The stool samples were scored from 1 to 7, on 7 consecutive days before the start of the study and on the last 7 days of the study. Then the scores were normalised to 4 (normal stool composition), so that the results represented the deviation from normal composition.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
4. Shotgun metagenome pipeline
For metagenome sequencing, the stool samples (~1 g) were collected according to GENIEUR Standard Operating Procedure.
For DNA isolation, ZymoBIOMICS 96 MagBead DNA kit was used. DNA concentration was measured by Qubit 3.0 fluorimeter.
Library preparation was performed by Nextflex Rapid XP SNA Seq kit. Library concentration was measured by Qubit HS DNA kit. Samples were sequenced on NextSeq2000 platform.
Trimmed reads were normalised by subsampling to the smallest sample size, for analysis with DRAGEN Metagenomics Pipeline.
Results were combined to a matrix by using KrakenTools. The resulting table was split by taxonomic groups (phylum, class, ordo, family, genus, species).
Dataset available in BioProject repository (ID: PRJNA885840)
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bód ayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
5. Statistical analysis
Paired t-test was used to determine the differences between the body composition variables and the blood parameters measured at baseline and at the end of the study within both groups. Unpaired t-test was used at the 2 different sampling times between the 2 groups.
Wilcoxon signed-rank test was used to determine the changes of bacterial amounts between the 2 groups and within the groups before and after the study period.
Linear correlation models were used to determine the correlation between the anthropometric features, body composition characteristics, blood parameters and amounts of different microbial taxonomic groups.
R statistical software packages were used for a further analysis, after normalisation of raw metagenome sequencing data
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6. Results
At the beginning of the study, no significant differences in body composition were found between the 2 groups. But at the end several changes were observed. However the only common change was the increased body weight.
CON group
INT group
In addition to body weight, fat-free mass, skeletal muscle mass, body cell mass, arm circumference, arm muscle circumference, protein mass and InBody score increased significantly. While the values of body fat mass, percent of body fat and visceral fat area decreased significantly.
In addition to body weight, body fat mass increased.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.1 Changes in gut microbioma
Results show that alpha diversity didn't change significantly during the study.
Firmicutes and Actinobacteria decresed significantly. Bacteroides increased significantly.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.1 Changes in gut microbioma
The Firmicutes/Bacteroidetes (F/B) ratio also changes significantly in both groups.
In order to obtain more information about the influence of the gut microbiome on the changes in athropometric parameters and body composition, researches aimed to identify the composition of gut microbiome at genus level and at species level. Then in the correlation analysis, the changes in the relative abundance of bacteria and their metabolic pathways were correlated with the changes in weight, body fat mass and skeletal mass.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.1 Changes in gut microbioma
Positive correlation:
- Relative abundance of acetate-producing bacteria with increased skeletal muscle mass (INT group)
- Reductive TCA pathway I with skeletal muscle mass (INT group)
- Pyruvate fermentation to propanoate I and both butyrate biosynthetic pathways with increased value of skeletal muscle mass (INT group)
- For the butyrate-producing species, changes in body fat mass with the relative abundance of the bacteria (INT group)
- Body fat mass with propionate-producing bacteria (CON group)
Negative correlation:
- Presence of acetate and propionate producer and reductive TCA cycle I pathway with skeletal muscle mass (CON group)
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.2 Changes in blood parameters
Positive correlation:
- Basal metabolic rate with the change in weight, skeletal muscle mass, body cell mass and fat-free mass value (INT group)
- Basal metabolic rate with fat-free mass, skeletal muscle mass and body cell mass (CON group)
- Body fat mass with percent and visceral body fat and arm circumference (CON group)
- Change in B12 level with hematocrit, the count of red blood cells and haemoglobin (CON group)
Negative correlation:
- Body fat mass with skeletal muscle mass (CON group)
- Haemoglobin with fat-free mass, skeletal muscle mass, body cell mass and basal metabolic rate (CON group)
- MCH with red blood cell distribution and skeletal muscle mass (CON group)
- Weight with the amount of red blood cells, hematocrit, haemoglobin and B12 level (INT group)
- Basal metabolic rate with red blood cell and haemoglobin (INT group)
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.3 Bristol stool test
At the end of the study, the INT group had a significant change in stool composition compared to the CON group (p=0.029), but the average stool composition remained in the healthy range.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Conclusions
Vegan protein supplementation improved body weight in 19 players, but skeletal muscle mass increased only in the INT group. They hypothesise that due to the additional intake of pre- and probiotics, fermentation of SCFA by the gut microbiome was more efficient in the INT group, which may have contributed to skeletal muscle development. The increased muscle metabolism is supported by the elevated MCH and MCHC levels in blood, which may improve the metabolic activities of muscle cells. Further studies with a larger number of participants are needed for a deeper understanding of the effects of protein and symbiotic supplements on muscle biosynthesis and overall homeostasis in elite athletes.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Thank you for your attention
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Gut microbiome composition: sports performance and protein absorpotion
Elena Cerofolini
Created on September 18, 2024
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Transcript
Gut microbiome composition: link between sports performance and protein absorption?
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
1. Introduction
A sufficient protein intake is essential for an adequate physical condition and athletic performance.Nowadays the study of the composition and the role of the gut microbiome in relation to sport nutrition have become an increasingly researched area.
This study aimed to examined how short-term diet of plant-based vegan protein affected muscle development in elite water polo players, in a 31-day period. It also investigated association between physical condition of the athletes and the composition of their gut microbiome after protein supplement with or without pre- and probiotics.(During the experiment none of the players were under any medical care, got any antibiotics or other treatments that could have influenced the microbiome composition)
INT group
CON group
(n=10)
(n=10)
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
2. Study design
INT group
CON group
They receive: 250 ml of Biotech vegan protein shake daily during the 31-day study
They receive: 250 ml of Biotech vegan protein shake daily during the 31-day study 15 drops of Herbaferm Probiotic solution, dissolved in 2 dl of water immediately after waking up 3.5 g of prebiotic mixture, homogenised daily in their 250 ml vegan protein shake
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
3. Analysis and data collection
Anthropometric and body composition analysis was performed by InBody 970, and several parameters were determined, such as weight, body fat mass, fat-free mass, skeletal muscle mass, percent body fat, visceral fat area, body cell mass, arm circumference, arm muscle circumference, protein mass, basal metabolic rate and overall fitness score (InBody Score).
Also blood samples were collected, and the parameter analysed were the count of red blood cells, value of haemoglobin, hematocrit, MCV, MCH, MCHC and vitamin B12 level.
The Bristol stool scale chart was used. The stool samples were scored from 1 to 7, on 7 consecutive days before the start of the study and on the last 7 days of the study. Then the scores were normalised to 4 (normal stool composition), so that the results represented the deviation from normal composition.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
4. Shotgun metagenome pipeline
For metagenome sequencing, the stool samples (~1 g) were collected according to GENIEUR Standard Operating Procedure.
For DNA isolation, ZymoBIOMICS 96 MagBead DNA kit was used. DNA concentration was measured by Qubit 3.0 fluorimeter.
Library preparation was performed by Nextflex Rapid XP SNA Seq kit. Library concentration was measured by Qubit HS DNA kit. Samples were sequenced on NextSeq2000 platform.
Trimmed reads were normalised by subsampling to the smallest sample size, for analysis with DRAGEN Metagenomics Pipeline.
Results were combined to a matrix by using KrakenTools. The resulting table was split by taxonomic groups (phylum, class, ordo, family, genus, species).
Dataset available in BioProject repository (ID: PRJNA885840)
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bód ayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
5. Statistical analysis
Paired t-test was used to determine the differences between the body composition variables and the blood parameters measured at baseline and at the end of the study within both groups. Unpaired t-test was used at the 2 different sampling times between the 2 groups.
Wilcoxon signed-rank test was used to determine the changes of bacterial amounts between the 2 groups and within the groups before and after the study period.
Linear correlation models were used to determine the correlation between the anthropometric features, body composition characteristics, blood parameters and amounts of different microbial taxonomic groups.
R statistical software packages were used for a further analysis, after normalisation of raw metagenome sequencing data
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6. Results
At the beginning of the study, no significant differences in body composition were found between the 2 groups. But at the end several changes were observed. However the only common change was the increased body weight.
CON group
INT group
In addition to body weight, fat-free mass, skeletal muscle mass, body cell mass, arm circumference, arm muscle circumference, protein mass and InBody score increased significantly. While the values of body fat mass, percent of body fat and visceral fat area decreased significantly.
In addition to body weight, body fat mass increased.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.1 Changes in gut microbioma
Results show that alpha diversity didn't change significantly during the study.
Firmicutes and Actinobacteria decresed significantly. Bacteroides increased significantly.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.1 Changes in gut microbioma
The Firmicutes/Bacteroidetes (F/B) ratio also changes significantly in both groups.
In order to obtain more information about the influence of the gut microbiome on the changes in athropometric parameters and body composition, researches aimed to identify the composition of gut microbiome at genus level and at species level. Then in the correlation analysis, the changes in the relative abundance of bacteria and their metabolic pathways were correlated with the changes in weight, body fat mass and skeletal mass.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.1 Changes in gut microbioma
Positive correlation:
- Relative abundance of acetate-producing bacteria with increased skeletal muscle mass (INT group)
- Reductive TCA pathway I with skeletal muscle mass (INT group)
- Pyruvate fermentation to propanoate I and both butyrate biosynthetic pathways with increased value of skeletal muscle mass (INT group)
- For the butyrate-producing species, changes in body fat mass with the relative abundance of the bacteria (INT group)
- Body fat mass with propionate-producing bacteria (CON group)
Negative correlation:Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.2 Changes in blood parameters
Positive correlation:
- Basal metabolic rate with the change in weight, skeletal muscle mass, body cell mass and fat-free mass value (INT group)
- Basal metabolic rate with fat-free mass, skeletal muscle mass and body cell mass (CON group)
- Body fat mass with percent and visceral body fat and arm circumference (CON group)
- Change in B12 level with hematocrit, the count of red blood cells and haemoglobin (CON group)
Negative correlation:Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
6.3 Bristol stool test
At the end of the study, the INT group had a significant change in stool composition compared to the CON group (p=0.029), but the average stool composition remained in the healthy range.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Conclusions
Vegan protein supplementation improved body weight in 19 players, but skeletal muscle mass increased only in the INT group. They hypothesise that due to the additional intake of pre- and probiotics, fermentation of SCFA by the gut microbiome was more efficient in the INT group, which may have contributed to skeletal muscle development. The increased muscle metabolism is supported by the elevated MCH and MCHC levels in blood, which may improve the metabolic activities of muscle cells. Further studies with a larger number of participants are needed for a deeper understanding of the effects of protein and symbiotic supplements on muscle biosynthesis and overall homeostasis in elite athletes.
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh
Thank you for your attention
Péter Fritza, Réka Fritzb, Pál Bódayc, Ádám Bódayd, Emese Batóe, Péter Kesserűf,g and Csilla Oláhh