Abstract

BackgroundObesity induced by a high-caloric diet has previously been associated with changes in the gut microbiota in mice and in humans. In this study, pigs were cloned to minimize genetic and biological variation among the animals with the aim of developing a controlled metabolomic model suitable for a diet-intervention study. Cloning of pigs may be an attractive way to reduce genetic influences when investigating the effect of diet and obesity on different physiological sites. The aim of this study was to assess and compare the changes in the composition of the gut microbiota of cloned vs. non-cloned pigs during development of obesity by a high-fat/high-caloric diet. Furthermore, we investigated the association between diet-induced obesity and the relative abundance of the phyla Firmicutes and Bacteroidetes in the fecal-microbiota. The fecal microbiota from obese cloned (n = 5) and non-cloned control pigs (n= 6) was investigated biweekly over a period of 136 days, by terminal restriction fragment length polymorphism (T-RFLP) and quantitative real time PCR (qPCR).ResultsA positive correlation was observed between body-weight at endpoint and percent body-fat in cloned (r=0.9, P<0.0001) and in non-cloned control pigs (r=0.9, P<0.0001). Shannon Weaver and principal component analysis (PCA) of the terminal restriction fragments (T-RFs) revealed no differences in the bacterial composition or variability of the fecal microbiota between the cloned pigs or between cloned and non-cloned control pigs. Body-weight correlated positively with the relative abundance of Firmicutes in both cloned (r=0.37; P<0.02) and non cloned-control pigs (r=0.45; P<0.006), and negatively with the abundance of Bacteroidetes in cloned pigs (r=−0.33, P<0.04), but not in the non-cloned control pigs.ConclusionThe cloned pigs did not have reduced inter-individual variation as compared to non-cloned pigs in regard to their gut microbiota in neither the obese nor the lean state. Diet-induced obesity was associated with an increase in the relative abundance of Firmicutes over time. Our results suggest that cloned pigs are not a more suitable animal model for gut microbiota-obesity related studies than non-cloned pigs. This study is the first to evaluate if cloned pigs provide a better animal model than conventional pigs in diet-intervention, obesity and gut microbiota research.

Highlights

  • Obesity induced by a high-caloric diet has previously been associated with changes in the gut microbiota in mice and in humans

  • A similar study based on 16S rRNA gene analysis indicated that bacterial community in human mono-zygotic twins (MZ) twins was slightly more similar than in unrelated individuals [3] suggesting that genetically identical individuals harbor a similar gut microbiota

  • computed tomography (CT) scanning of body fat showed that obese non-cloned control pigs had a higher average percentage of body-fat (41.1±1.3%) than obese cloned pigs (28.4 ± 2.3%, P=0.004)

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Summary

Introduction

Obesity induced by a high-caloric diet has previously been associated with changes in the gut microbiota in mice and in humans. The aim of this study was to assess and compare the changes in the composition of the gut microbiota of cloned vs non-cloned pigs during development of obesity by a high-fat/ high-caloric diet. In studies of diet-induced obesity and its association with the gut microbiota, it may be preferable to eliminate the influence of host genotype on the composition of the gut microbiota by choosing genetically identical animals. In a more recent study on the relationship between gut microbiota, diet and genetic influences in mice, the authors stated that the changes in gut microbiota were unrelated to genetically induced obesity and were merely due to high-fat (HF) diet [4]. The influence of the host genome on the gut microbiota currently remains controversial

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