Colonic luminal microbiota and bacterial metabolite composition in pregnant Huanjiang mini-pigs: effects of food composition at different times of pregnancy
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OPEN
received: 06 July 2016
accepted: 26 October 2016
Published: 05 December 2016
Colonic luminal microbiota and
bacterial metabolite composition
in pregnant Huanjiang mini-pigs:
effects of food composition at
different times of pregnancy
Xiang-feng Kong1,2,*, Yu-jiao Ji1,*, Hua-wei Li1, Qian Zhu1, F. Blachier3, Mei-mei Geng1,
Wen Chen1 & Yu-long Yin1,2
The gut harbours diverse and complex microbiota, which influence body health including nutrient
metabolism, immune development, and protection from pathogens. Pregnancy is associated with
immune and metabolic changes that might be related to microbiota compositional dynamics. We
therefore investigated the colonic luminal bacteria community in Huanjiang mini-pigs fed diets
with different nutrient levels from the first to third trimester of pregnancy. The concentrations of
intestinal metabolites including short-chain fat acids, NH3-N, indole, skatole, and bioamines were also
determined. We found that the colonic bacteria species richness estimators (Chao1 and ACE) decreased
with increased gestational age. The dominant phyla identified were Firmicutes and Bacteroidetes;
the dominant genera were Lactobacillus, Treponema, Ruminococcus, Clostridium, and Prevotella.
In addition, microbiota displayed spatial and temporal heterogeneity in composition, diversity,
and species abundance in different colonic segments from the first to third trimester of pregnancy.
Furthermore, the bacterial metabolites also changed according to the diet used and the pregnancy
stage. These findings suggest that colonic bacteria richness decreased as gestational age increased,
and that the higher nutrient level diet increased the production of metabolites related to nitrogen
metabolism. However, although the higher nutrient diet was associated with pregnancy syndrome,
causal links remain to be determined.
Dietary nutrients during gestation play important roles in the reproductive performance of sows and within-litter
variation of their offspring1. It was initially thought that sows in early gestation should receive large nutrient
amounts2. However, high nutrient levels during gestation can increase sow fat mass, resulting in farrowing problems, poor dietary intake during lactation, and less reproductive performance in the next cycle3. Notably, sows fed
diets with high energy content during early gestation suffer embryo deaths4. Conversely, after mating, nutrition
restriction supports progesterone rise and embryonic survival in gilts. Thus, excess or inadequate dietary energy
and protein ingestion by pregnant pigs can contribute to poor reproductive performance and impaired physiological functions.
Uterine growth during gestation causes profound gastrointestinal tract modification, giving rise to altered
gastric motility and intestinal transit time along with gastrointestinal disturbances5. Recently, Koren et al. demonstrated that in humans, the gut microbiota composition during first trimester pregnancy is very similar to that
of a healthy non-pregnant control group but becomes enriched in Proteobacteria and Actinobacteria during the
third trimester6. Notably, these latter changes are similar to those detected in inflammatory bowel disease and
1
Key Laboratory of Agro-ecological Processes in Subtropical Region, Hunan Provincial Engineering Research Center
for Healthy Livestock, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Hunan 410125, China.
2
Research Center of Mini-pig, Huanjiang Observation and Research Station for Karst Ecosysterm, Chinese Academy
of Sciences, Huanjiang, Guangxi 547100, China. 3INRA, CNRH-IdF, AgroParisTech, UMR 914 Nutrition Physiology
and Ingestive Behavior, Paris 75005, France. *These authors contributed equally to this work. Correspondence and
requests for materials should be addressed to X.-f.K. (email: ) or Y.-l.Y. (email: )
Scientific Reports | 6:37224 | DOI: 10.1038/srep37224
1
www.nature.com/scientificreports/
NRC diet
Items
P values
CNF diet
45 d
75 d
110 d
45 d
75 d
110 d
SEM
Diet
Stage
D*S
Row tags
41039.2
41118.7
42682.7
41276.6
42296.3
40799.0
268.06
0.787
0.602
0.140
Effective Tags
39920.6
39569.7
41271.3
40541.8
41110.1
39401.7
281.97
0.872
0.983
0.110
Number of OTUs
1209.8
1002.0
1024.3
1145.9
1161.3
1036.7
21.84
0.377
0.017
0.040
Chao1
1342.5
1118.6
1146.7
1305.2
1304.0
1142.2
24.66
0.285
0.009
0.055
ACE
1356.7
1326.6
1165.1
1317.2
1326.6
1165.1
13.59
0.257
0.008
0.037
Simpson
0.98
0.95
0.94
0.92
0.93
0.92
0.01
0.088
0.642
0.556
Shannon
7.42
6.56
6.52
6.81
6.75
6.70
0.13
0.768
0.223
0.342
Coverage
0.993
0.994
0.993
0.993
0.993
0.994
<0.01
0.132
0.010
0.073
Row tags
42545.2
41068.3
41185.0
41299.1
41226.0
40558.7
289.32
0.368
0.352
0.567
Effective Tags
41552.6
39506.7
39747.7
40266.6
39892.1
38992.7
296.38
0.384
0.103
0.442
Number of OTUs
1224.6
1101.7
1101.7
1183.4
1221.3
1101.3
16.53
0.419
0.062
0.063
Chao1
1372.5
1235.8
1248.0
1332.3
1360.7
1217.4
17.95
0.601
0.038
0.062
ACE
1392.0
1252.8
1255.3
1348.9
1388.1
1237.4
18.19
0.469
0.031
0.044
Simpson
0.98
0.98
0.96
0.97
0.97
0.97
<0.01
0.634
0.490
0.632
Shannon
7.52
7.23
6.86
7.29
7.37
7.34
0.10
0.553
0.593
0.470
Coverage
0.992
0.993
0.993
0.993
0.992
0.994
<0.01
0.922
0.066
0.076
Proximal colon
Distal colon
Table 1. Alpha diversity indices of colon bacterial communities in Huanjiang mini-pigs at different stages
of pregnancy.
obesity. In the gut, the bacteria load also increases over the course of gestation7. However, Jost et al. reported little
or no modifications of the faecal microbiota composition during early and late pregnancy8. In humans, disorders
associated with gut microbiota dysbiosis include obesity9, type 2 diabetes10, and malnutrition11. The mammalian
gut, especially the hindgut, is colonised by a large number of microbes with varying microbial composition in the
different parts of the gut12. The colon is colonised by up to almost 1012 bacteria per gram digesta13. The microbiota
in the gut lumen plays an important role in the digestion and absorption of nutrients, prevention of pathogen
colonisation, and maintenance and regulation of normal mucosal immunity14. Recently, the roles of gut microbiome activity and their metabolites in regulating host physiological functions in association with longevity15 and
the metabolic changes during pregnancy6 have attracted considerable interest. Intestinal microbiota composition
is affected by multiple factors including environmental parameters, dietary composition in terms of quality and
quantity, and antibiotics use16. Many dietary components including protein and amino acid-derived bacterial
metabolites such as short-chain fatty acids, NH3-N, indole, skatole, and bioamines affect the relationship between
gut microbiota and the intestinal mucosa, and this relat (...truncated)
