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R package version 1. Google Scholar. Download references. The author would like to thank Dr. Qiuwei Pan, department of Gastroenterology and Hepatology, Erasmus MC, for constructive criticism of the manuscript. China-Malaysia National Joint Laboratory, Biomedical Research Center, Northwest Minzu University, Lanzhou, China.

Department of Basic Medicine, Medical School, Kunming University of Science and Technology, No. Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Center, Rotterdam, The Netherlands.

You can also search for this author in PubMed Google Scholar. ZL and JHS conceived the study and together with LHL and YYW further finalized the design.

LHL, YXS, FLL and YYW planned the laboratory procedures. LHL, YYW and JHS conceptualized the statistical analyses. LHL, YYW, YXS and FLL performed sample collection.

All authors, in particular ZRM, contributed to interpretation of the data. All authors contributed to and approved the final manuscript.

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Download PDF. Abstract Background It has recently been reported that intermittent fasting shapes the gut microbiota to benefit health, but this effect may be influenced to the exact fasting protocols.

Conclusions These data indicated that intermittent fasting shapes gut microbiota in healthy mice, and the length of daily fasting interval may influence the outcome of intermittent fasting.

Background Gut microbiota consists of a group of microorganisms that live in the mammalian intestinal tract and plays key roles in health and disease.

Full size image. Table 1 Microbiome alpha diversity indices between different time durations at the end of fasting or 1 month after the cessation of intermittent fasting Full size table.

Table 2 Analysis of beta diversity of gut microbiota by ANOSIM and Adonis test Full size table. Discussion Fasting, in particular intermittent and timing fasting, is being widely practiced for various purposes in global population, for at least millennia now.

Next-generation sequencing Fecal DNA extraction and next-generation sequencing of 16S ribosomal RNA gene amplicons were performed by NoveGene, as reported elsewhere [ 33 ]. Abbreviations hrs: Hours CTR: Control ANOSIM: Analysis of similarities Adonis: A multivariate ANOVA based on similarity tests PCoA: Principal coordinates analysis LDA: Liner Discriminate Analysis LEfSe: Liner Discriminate Analysis LDA coupled with effect size measurements OTUs: Operational taxonomic units OTU: Operational taxonomic unit.

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Numerous research studies find out that IF mostly results in weight loss and improvement of blood profile Aloui et al. In this study, it was found that IF has an impact on body mass indexes, blood lipid profiles, and gut microbial profiles.

Surprisingly, weight increase was observed in underweight women. These results show that this IF regimen tends to normalize the total body mass without its unwarranted depletion in healthy individuals.

Reduction of body weight by IF has been reported Adlouni et al. The number of Bifidobacteria and Lactobacillus increased in all five groups, and it is well established that reduced body weight is linked to an increasing trend in gut microbial diversity, and overweight or obesity is linked to reduced gut bacterial diversity Ley et al.

In our study, the weight reduction was found linked to an increasing trend in bacterial diversity, while a decreasing trend in bacterial diversity was observed especially in women those gained weights.

In an earlier report, the impact of IF on a female subject showed an increase in weight was due to the use of energy-dense diets and less physical activities during the fasting days Sadeghirad et al.

However, the impact of IF is not gender-specific, when tested in the mice model Piotrowska et al. Abdelgadir in conducted a study on three different nations and reported that people using normal diets with physical activities have improved lipid profiles after IF while people eating more or energy-rich diets or doing no or limited physical activities have no change in their lipid profile at the end of study Abdelgadir et al.

Yeoh et al. The total cholesterol level decreased in all female volunteers. IF lowered total aerobic bacterial count in all the male and female volunteers when checked through conventional culturing methods. The number of total Lactococcus and total Lactobacillus , when assessed through culturing method, seen increased at the end of the IF trial in both male and female volunteers.

Recently, a study established the link between gut bacterial diversity and lipid blood profile, which showed that fasting sugar level, total triglycerides, total cholesterol, and low-density lipoproteins are associated with reduced gut bacterial diversity as compared to the control cohort Rebolledo et al.

Another study conducted on Finnish-men also showed that elevated blood lipids are associated with increased gut bacterial diversity Org et al.

Yang et al. The in vitro cholesterol assimilation activity of gut microbiota isolated from healthy individuals has been confirmed by different researchers recently, and the presence of bile salt hydrolase bsh genes is reported in Lactobacillus and Bifidobacteria , mostly used for cholesterol assimilation activities Pereira and Gibson, ; Begley et al.

These bacteria are known for anti-inflammatory activities, and inulin is the key component that favorably increases the number of Bifidobacteria spp de Souza et al. These findings favored our results where all the samples carried Lactobacillus spp while their lipid profile was also positively improved.

It has been demonstrated that overweight individuals carry a high microbial load in their intestines when compared with normal ones Mokkala et al. Another study showed that increased bacterial richness is associated with parasite-positive guts Andersen et al.

It was explained by Maine Margo that increased diversity in the intestine is considered a healthy flora.

They have more potential to degrade and metabolize diverse nutrient components Maine and Kelly, Therefore, the reduction in bacterial richness and increase in diversity confirm a positive alteration in the intestinal bacteria.

Changes were seen in the percentages of bacterial phylum at the end of the study Figure 8. Normal weight male group had a higher percentage of Proteobacteria Increase in Actinobacteria from 0. Male underweighted group before IF had the highest Firmicutes of The Actinobacteria also reduced from In female underweighted group, Firmicutes and Bacteroides decreased from In female normal group, no significant changes at phylum level were observed.

It has been concluded that too high and too low Proteobacteria are signs of dysbiosis. Figure 8. Hierarchically clustered heatmap of the bacterial distribution of different communities.

Double hierarchical dendrogram shows the bacterial distribution. The bacterial phylogenetic tree was calculated using the neighbor-joining method, and the relationship among samples was determined by Bray—Curtis distance and the complete clustering method.

The heatmap plot depicts the relative percentage of each bacterial phylum variables clustering on the vertical axis within each sample group horizon-axis clustering. The relative values for bacterial phylum are indicated by the color intensity with the legend indicated at the bottom of the figure.

This bacterium is isolated from grasses and is kept in risk group one, and it may cause infection in humans Söhngen et al. Later, at the end of the study, it decreased to 0. A study conducted by Cucchi et al.

Participants consumed a vegetable-rich diet during the study period. The Ruminococcus flavefaciens are fiber digesting bacteria mainly involved in interactive digestion and acquisition of the nutrients in the intestine Cucchi et al. Ejtahed et al. In another study the effect of energy restriction on body weight and gut microbiota was evaluated.

It was concluded that an increase in the number of Bifidobacterium sp and lactobacillus spp is associated with weight loss Santacruz et al. This species is heterofermentative and has a broad range of substrate utilization, thus providing more nutrients to their host Kageyama and Benno, The Collinsella aerofaciens was associated with the prevention and cure of irritable bowel diseases Kassinen et al.

These species have been used for the treatment of type 2 diabetes Wang and Jia, Less number of Dorea spp was observed, and it is known that an increased number of Dorea spp can contribute to the development of irritable bowel disease Rajilić-Stojanović et al.

These findings present an improvement in the health status of the intestine and the whole host. In the male underweighted group, evenness and bacterial diversity were increased after the trial. Studies have shown that Campylobacter spp are normally present in animals and poultry, and their count goes high in warmer months.

Not all Campylobacter spp are pathogenic to humans Keener et al. The Campylobacter spp. have shown the colonization of the large intestine Saint-Cyr et al. This study was performed in warmer months, while the volunteers were consuming poultry meat as a part of their routine diet.

This bacterium is responsible for the treatment of metabolic acidosis Marx et al. The Oscillospira spp. were decreased in the groups. According to a study, a decrease in Oscillospira spp is considered anti-obesity signature bacteria Walters et al.

Increased Roseburia spp. are associated with gall stone formation Tamanai-Shacoori et al. Strength of the study: Blood lipid profile was improved and modulated gut microbiota even though the volunteers were taking their ad libitum diets during the eating windows of the experiment and were living their routine life.

No dietary or living habits were advised to be changed or restricted, but, in many gut modulation studies, experimental subjects are passed through dietary restriction or changing living styles. Recently, a study conducted on a mice model for the improvement of blood lipid profile and modulation of gut microbiota kept the experimental mice restricted to a diet having the main portion of isoquercetin and inulin Tan et al.

In another study, antibiotics were administered in the diet for the improvement of blood lipid profile and modulation of gut microbiota Rune et al. We have conducted our experimental trial on the human population, and it reflects the actual outcome and can be used as a reference for further studies, while in many cases, such studies are performed on mice model.

There is a difference in the gut microbiota of humans and mice which affects the outcome of any experimental trial. Recently, the gut microbiota of mice and humans was compared using large datasets, and it was found that both the gut microbiota are similar at phylum level but have large variations at the species level Xiao et al.

The core microbiota of mice is small, while the core of human gut microbiota is large Kostic et al. The gut microbiota of mice can be changed by the diet within 1 week, while human gut microbiota does not change so quickly Wang and Jia, The difference between the results of women and men may be due to the fasting days, and women were exempted to fast during their menstrual cycle.

The number of fasting days is not defined yet. Limitation of the study: The number of male and female samples was not equal and thus may have an effect on the results.

This may have influenced the results. The datasets presented in this study can be found in online repositories. The studies involving human participants were reviewed and approved by BEC-FBS-QAU MI and MIR conceived the study and experimental designing.

MI supervised the study. AA and MI analyzed the data and critically revised the manuscript. MYK contributed to sample analysis. MNK and SIK conducted experimental work and drafted the manuscript. All authors contributed to the preparation of the final manuscript.

We would like to acknowledge Hafiz Abrab Sikandar for his guidance and all the volunteers of the study who participated willingly.

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers.

Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher. Supplementary Figure 1 Informed consent form for participants.

Supplementary Figure 2 Diet and living habits of Participants. Supplementary Figure 3 Impact of intermittent fasting on culturable aerobic bacteria and fungi from fecal material of participants. Supplementary Table 1 Impact of intermittent fasting of serum lipid profile of male participants.

Supplementary Table 2 Impact of intermittent fasting of serum lipid profile of female participants. Supplementary Table 3 Impact of intermittent fasting of gut microbiota at phylum level of participants.

Supplementary Table 4 Impact of intermittent fasting of gut microbiota at genera level of obese female participants. Supplementary Table 5 Impact of intermittent fasting of gut microbiota at genera level of normal weight female participants.

Supplementary Table 6 Impact of intermittent fasting of gut microbiota at genera level of under-weight female participants. Supplementary Table 7 Impact of intermittent fasting of gut microbiota at genera level of normal weight male participants.

Supplementary Table 8 Impact of intermittent fasting of gut microbiota at genera level of under-weight male participants. Abargouei, A.

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However, there is significant variability in this capacity among different species and strains Vacca et al. Thus, this could explain the difference observed in various studies. However, in our study, although there was no statistically significant correlation between the Lachnoclostridium levels and the total fiber intake, it was observed that both the total fiber intake and the abundance of Lachnoclostridium were decreased.

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It has also been reported that a higher intake of polysaccharides and plant protein leads to a higher abundance of Coprococcus and other butyrate-producing bacteria Garcia-Mantrana et al. Although the plant protein intake decreased in this study, it was not statistically significant.

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This correlation was linked to unhealthy dietary habits and obesity Takada et al. There is currently no research in the literature that demonstrates a relationship between Fusicatenibacter and nutrient consumption.

It has been reported that consuming refined sugar can influence both the function and composition of the intestines. They stated that further studies are necessary to confirm this effect. Our research did not detect any relationship between dietary patterns and Akkermansia. Consuming whole grains plays a role in gut microbiome modulation Martínez, although no evidence of gut microbiota shift from grain consumption was found in our study.

Intestinibacter of Firmicutes phylum was positively correlated with protein-derived energy. Our study found no significant correlation between protein intake and Intestinibacter. However, we did observe a negative correlation between Intestinibacter and the consumption of nuts.

In this study, in which we evaluated the effects of RIF during the standard fasting period, it was very difficult to evaluate the differences in interindividual diet patterns since they did not follow a standard dietary pattern.

Using a standard dietary pattern in future studies will aid in effectively evaluating the results. This research has several limitations. First, the sample size is limited because it was challenging to find individuals that met the inclusion criteria.

The small sample size makes it essential to do more research on the impacts of dietary practices like fasting on a greater study cohort. Further comprehensive and prolonged research is necessary to evaluate the complete influence of RIF on gut microbiota modulation.

Another limitation of this study is sleep and changes in dietary intake during RIF. Sleep restriction could alter gut microbiota composition because RIF is associated with sleep duration and nighttime sleepiness. Therefore, sleep may be a confounding factor in the alteration of gut microbiota.

In this study, although there was no change in the amount of energy, carbohydrate, and protein taken in the diet, the dietary fat and water-soluble fibers were different.

Therefore, differences in dietary intakes may also be a confounding factor in gut microbiota change. In conclusion, diet and fasting have important effects on the gut microbiota. These effects differ between individuals. During Ramadan, although the duration of fasting is similar, there are significant differences between the foods consumed by individuals.

The effects of these consumed foods are the basis of the different changes in the gut microbiota. Therefore, future studies must evaluate the effects of people following similar dietary patterns during Ramadan.

The datasets presented in this study can be found in online repositories. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.

DS contributed to the design and implementation of the research, to the analysis of the results and to the writing of the manuscript. GAC contributed to the design and implementation of the research. ES designed and performed data analysis, prepared figures, and contributed to writing and reviewed the manuscript.

BE and US have contributed to writing and reviewing of the manuscript. MB has contributed to design of the research, write, and review of the manuscript.

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers.

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Improvements were seen for both groups — those who fasted and those who focused on reducing their daily calorie intake. The analysis suggested that a person can improve the diversity of their microbiome and potentially their overall health using the weight-reduction strategy of their choice.

The new study reinforces the idea that changes in gut bacteria occur during weight loss. The researchers observed several associations between the abundance of microbes associated with metabolism and obesity and DNA methylation, a process by which gene regulation is altered, potentially impacting our health.

The study appears in Nutrients. Inside the human body are roughly trillion symbiotic microbial cells. Most of these are bacteria, and most live in the upper and lower intestine. Our understanding of these tiny organisms is still somewhat in its infancy.

However, it is clear that they are influential actors in our health. Gastroenterologist Dr. It mediates any type of inflammatory process going on within your body. Inflammation in the body has been implicated in many medical issues, from cancer to diabetes , dementia , and heart disease.

In addition, the microbes in the microbiome influence other processes as well, including appetite and obesity. You want a very diverse microbiome in order to decrease and regulate all the mechanisms within your body. Research bears out the value of a diverse microbiome.

The researchers, following their earlier analysis , had commented that the mechanism could be the benefits seen with changes in metabolism, weight loss, cardiometabolic factors, or even improvements in dietary patterns associated with the two arms of intervention.

Bedford suggested a simpler reason. The dietitian also expressed concern that fasting diets and calorie reduction could cause further harm to people with a history of disordered eating. Fasting can be performed in a variety of ways.

While participants in the study fasted 3 days a week, fasting can also be done for a few hours, or for multiple days in a row. He cautioned that fasting is not a good idea for people with diabetes, since the prolonged lack of food causes fluctuations of blood sugar and insulin levels.

Previous research has found that calorie reduction, if it is too extreme, can cause an increase in pathogenic bacteria in the gut, and may otherwise disrupt the microbiome. Bedford did not question the findings of this research.

However, he suggested that extreme calorie reduction is an unlikely practice. It takes an enormous amount of discipline to do that. And in terms of plants, again, a very limited number of plant products that we also consume.

He noted the existence of so-called blue zones , regions around the world in which people live exceptionally long lives. And it changes the microbiome for the better, and therefore the less disease, fewer issues, and fewer problems. There is a lot of hype around intermittent fasting, but what are its actual benefits, and what are its limitations?

We lay bare the myths and the…. Can we use food and diet as medicine? If so, to what extent? What are the pros and cons of this approach to healthcare? PCR amplification was carried out in a reaction mixture containing Phusion® High-Fidelity PCR Master Mix New England Biolabs, Ipswich, MA, USA.

Finally, the library was sequenced on an Illumina platform. Merged reads were quality checked by using split-libraries-fastq.

py in QIIME ver 1. Using the UCHIME algorithm, the reads were compared with the reference database SILVA database to remove chimera sequences. Representative sequences were classified against the SILVA v reference taxonomy using a negative Bayesian classifier implemented within mothur [ 38 , 39 ].

Finally, a rarefied feature table was created at one depth of sequence per sample, and all of the downstream analyses were performed with this rarified OTU operational taxonomic unit table.

Alpha diversity indices Shannon diversity and Simpson index were calculated using alpha-diversity. py in QIIME. Beta diversity was computed using Bray-Curtis distance metrics. Multivariate data analysis methods of Adonis nonparametric manova and ANOSIM analysis of similarities were used to identify whether the daily fasting duration had an effect on the microbial communities.

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Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. R package version 1. Google Scholar. Download references. The author would like to thank Dr. Qiuwei Pan, department of Gastroenterology and Hepatology, Erasmus MC, for constructive criticism of the manuscript.

China-Malaysia National Joint Laboratory, Biomedical Research Center, Northwest Minzu University, Lanzhou, China. Department of Basic Medicine, Medical School, Kunming University of Science and Technology, No.

Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Center, Rotterdam, The Netherlands. You can also search for this author in PubMed Google Scholar.

ZL and JHS conceived the study and together with LHL and YYW further finalized the design. LHL, YXS, FLL and YYW planned the laboratory procedures.

LHL, YYW and JHS conceptualized the statistical analyses. LHL, YYW, YXS and FLL performed sample collection. All authors, in particular ZRM, contributed to interpretation of the data.

All authors contributed to and approved the final manuscript. Correspondence to Junhong Su. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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