- Research article
- Open Access
Intestinal microbiota could transfer host Gut characteristics from pigs to mice
- H. Diao†1,
- H. L. Yan†1,
- Y. Xiao†1,
- B. Yu1,
- J. Yu1,
- J. He1,
- P. Zheng1,
- B.H. Zeng2,
- H. Wei2Email author,
- X.B. Mao1Email author and
- D.W. Chen1Email author
© The Author(s). 2016
- Received: 24 March 2016
- Accepted: 27 September 2016
- Published: 11 October 2016
The Erratum to this article has been published in BMC Microbiology 2016 16:253
The present study was conducted to compare the differences in gut microbiota composition and gut-phenotypes among pig breeds, and determine whether these differences would transmit to mice colonized with fecal microbiota of different pig breeds. A total of 24 1-day-old germ-free BALB/C mice were divided into 3 groups (TFM, YFM and RFM), which were transplanted with intact fecal microbiota of Tibetan pig (TP), Yorkshire pig (YP) and Rongchang pig (RP), respectively.
Results showed that different pig breeds exhibited distinct gut microbiota profile based on high-throughput pyrosequencing. YP exhibited a lower Firmicutes/Bacteroidetes ratio and apparent genera differences compared with RP and TP, and higher levels of bacteria from Spirochaetes were observed in TP compared with RP and YP (P < 0.05). Transplanted porcine microbiota into GF mice replicated the phenotypes of pig donors. Moreover, the three groups of donor pigs and their mice recipients exhibited different intestinal index and morphology. TP and RP had higher intestinal weight and relative CDX2 mRNA expression in ileum than YP, and longer intestine, higher villus height of duodenum and jejunum were observed in TP compared with YP and RP (P < 0.05). TP exhibited higher GLP-2 mRNA expression in duodenum and jejunum than RP (P < 0.05). Similarly, YFM had lower intestine weight and CDX2 mRNA expression in ileum than TFM and RFM (P < 0.05). The intestine length in TFM was longer than that in RFM, and TFM had higher villus height in duodenum and jejunum and GLP-2 mRNA expression in ileum than the other two groups (P < 0.05). Besides, the digestive and absorptive ability was different among the three groups in donor pigs and mice recipients. YP had higher jejunal lactase and maltase activities than TP and RP, while TP had higher activities of jejunal ATPase, γ-GT, and relative SGLT1 mRNA expression in duodenum and jejunum than YP and RP (P < 0.05). Likewise, YFM had higher jejunal sucrase and maltase activities than TFM and RFM, whereas higher jejunal γ-GT activity and relative SGLT1 mRNA expression in duodenum and ileum were observed in TFM compared with YFM and RFM (P < 0.05). In addition, Tibetan pigs-derived microbiota improved gut barrier in mice recipients. The concentration of MDA in YP was higher than that in TP and RP (P = 0.078), and the relative ZO-1 mRNA expression in ileum in TP was higher than that in YP (P < 0.05). Likely, compared with TFM and RFM, YFM exhibited increasing MDA concentration in jejunum (P = 0.098), and the relative ZO-1 mRNA expression in duodenum and ileum in TFM were higher than that in YFM (P < 0.05).
There were huge differences in gut microbiota composition and gut characteristics among pig breeds, and gut microbiota could partially convey host gut characteristics from pigs to mice.
- Pig breeds
- Intestinal development
- Digestive and absorptive activities
- Germ-free mice
- Fecal microbiota transplantation
Gut microbiota is a category of microorganisms inhabiting the mammalian gastrointestinal tract and participates in most host’s physiological processes from nutritional status to behavior and stress response . Indeed, a broad range of biological functions that the host couldn’t otherwise accomplish are performed by this special organ, including providing nutrients, modulating gastrointestinal development and shaping immune system . The commensal microbiota is highly variable from individual to individual, and the different regions of the host intestine harbor specific microbial communities altering in density and diversity . Accumulating evidence indicates that diet, environment, host genotypes and phenotypes can strongly influence the composition of the gut microbiota in various mammals [4–7]. In the studies of monozygotic or dizygotic twin pairs, huge differences in the gut microbiota profiles were found between healthy twins and twins with malnourishment or obesity [8, 9], and these modifications can also be brought about by adding or deleting one gene to a model host organism [10, 11]. These researches strengthened the concept that distinct gut microbiota compositions are existed in host with different genotypes or phenotypes. The Tibetan pig (TP) is a typical native pig breed from Tibetan plateau, and is characterized by stronger adaptability and disease resistance . The Rongchang pig (RP) is also a typical native pig breed from China, and is characterized by better meat quality . While the Yorkshire pig (YP) is an imported breed and is characterized by high growth rate, elevated lean meat percentage and inferior meat quality [14, 15]. Several previous researches in large domestic animals also demonstrated that the profile of gut microbiota showed large distinction between breeds [16–18]. Thus, we speculated that there are huge differences in the gut microbiota compositions among TP, YP and RP.
Recently, there has been a considerable increase in the study of gut commensal flora and local host-microbe interactions. Host-microbe interactions occur initially along the surfaces of mucosa, and the intestinal mucosa contributes the largest surface area within the body, which separates the lumenal digesta from the internal milieu through a single layer of epithelium and represents a major surface for microbial colonization . The overall balance in the composition of the gut microbial communities is important in ensuring homeostasis at the intestinal mucosa. Comparisons of germ free animals and conventional raised animals reveal the vital role of gut microbiota in the structural and functional development of the gastrointestinal tract [20–23]. A better intestinal morphology is beneficial for nutrients digestion and absorption. Moreover, gut barrier integrity can be improved by the preferable capacity of digestion and absorption in host . Currently, fecal microbiota transplant (FMT) technology and the human flora-associated (HFA) animal model based on germ-free animals are commonly used in the research of the relationship between gut microbes and the human phenotype [24, 25]. Studies of transplanting fecal microbota from healthy human to those with various diseases (such as inflammatory bowel disease, clostridium difficile enterocolitis, metabolic phenotype and disease, immune disorder) have indicated that it is crucial to recover the normal microbial composition of the host during treatment for these diseases [26–29]. However, whether the gut properties are transmissible among animal species via fecal microbiota transplantation remains unclear. A study conducted by Zhang (2008) found that the allometry value of small intestine in Tibetan pig was smaller than that in foreign breeds . Therefore, we hypothesized that the differences in gut characteristics among pig breeds can be transferred from the pig donors to germ-free (GF) mouse recipient.
The objective of present study was conducted to explore differences in the gut microbiota profiles and phenotypes in terms of intestinal morphology, digestive and absorptive capacity and barrier among TP, YP and RP and their mice recipients, and to determine whether the differences in the gut characteristics are transmissible via fecal microbiota transplantation. Thus, the relative achievements of this study will provide evidences for further understanding interactions among microbiota and host gut characteristics, interpreting the possible mechanism of gut microbiota regulating host gut development, and illuminating the possible way manipulating gut health by utilizing gut microbiota as the key target.
The 5 Yorkshire pigs, 5 Tibetan pigs and 5 Rongchang pigs (12 weeks of age) were used in this experiment as fecal donors. All pigs were provided by reservation farms for these three pig breeds. TP, YP and RP were housed separately in individual metabolic cages in three environmentally controlled rooms on our experimental farm for 8 weeks until sacrifice, in which pigs were allowed ad libitum access to water and diet. A corn-soybean diet was formulated according to NRC (2012) requirements and Chinese feeding standards for local pigs (2004).
Germ-free BALB/C mice were provided by the Department of Laboratory Animal Science of the Third Military Medical University. A total of 24 1-day-old germ-free BALB/C mice, which were maintained in sterile Trexler plastic film isolators (Fengshi Laboratory Animal Equipment, China) and housed in polycarbonate cages on sterile wood chips at 22 ~ 24 °C at a relative air humidity of 45 ~ 55 % on a 12-h light–dark cycle, were used as recipients for fecal microbiota transplantation in this study. 1-day-old mice were breast fed by the germ free foster mice before weaning, and then were fed ad libitum with a chow diet sterilized by 60Co gamma radiation after they were weaned.
Fecal microbiota transplantation and treatments
On the basis of the standard for donor identification and screening described by Hamilton et al., pigs used in the present study did not have diarrhea or other digestive disorders, never received medication before the study, and were fed a diet without antibiotics and probiotics for at least 2 month before feces collection . The fresh feces of all the pigs were collected separately after 12-h fasting. In order to acquire representative fecal material for each breed, feces samples of each breed were mixed and then used as fecal inoculum. The remaining feces of each pig were stored at −80 °C until DNA extraction.
The stool suspension was prepared as described by Zeng et al. (2013) . In brief, 1:9 (w/v) sterile prereduction phosphate buffer (0.1 mol/L, pH7.2) was added into the mixed fresh feces. The suspension was mixed and passed through 2.0, 1.0 and 0.5 mm stainless steel laboratory sieves to remove larger particles, and then stored at −80 °C until fecal transplantation.
Newborn germ-free mice in each treatment were infused by intragastric gavage with 0.05 mL fecal suspension of Tibetan, Yorkshire or Rongchang pigs, and 2 mL aliquots were spread on the fur of each germ-free foster mouse. These mice were maintained in the same manner as germ-free mice. Three treatment groups were: 1) Tibetan porcine flora-associated mice (TFM), 2) Yorkshine porcine flora-associated mice (YFM), and, 3) Rongchang porcine flora-associated mice (RFM). Each group contained 8 mice. The study lasted for 5 weeks.
Sample collection of donors and recipients
Mouse pups were acquired by provoking defecation through slightly pushing in the lower abdomen using a moist cotton swab on week 5. All fecal samples were immediately stored at −80 °C until DNA extraction. All pigs were sacrificed at 20 weeks of age by using intravenously administrated dose of chlorpromazine hydrochloride (2 mg/kg body weight) as anesthetics. All mice were sacrificed at 5 weeks of age, and fasted overnight on the day before being killed by cervical dislocation. The length and weight of the small intestine and large intestine were measured, and the tissues of duodenum, jejunum and ileum were immediately isolated and preserved in 4 % paraformaldehyde solution. In addition, the tissues of duodenum, jejunum and ileum were immediately collected and stored at −80 °C.
16S rRNA amplicon sequencing
Total DNA was isolated using the QIAamp DNA stool Mini Kit (Qiagen, GmbH Hilden, Germany). The concentration and purity of extracted genomic DNA were measured using a NanoDrop ND-1000 Spectrophotometer (NanoDrop, Germany). The integrity of extracted genomic DNA was determined by electrophoresis on 1 % agarose gels. Sequencing and bioinformatics analysis were performed by BGI (Shenzhen, China). DNA library was prepared before high-throughput sequencing as previously described . The resulting sequences were clustered to operational taxonomic units (OTU) using USEARCH drive5 at 97 % sequence identity. Relative abundance of each OTU was examined at different taxonomic levels.
Histology of intestine
The duodenal, jejunal and ileal histomorphology were determined as described previously . Briefly, following the fixing, the segment of the jejunum was excised, dehydrated and embedded in paraffin. Then, consecutive sections (5 μm) were cut and stained with haematoxylin and eosin. The villus height and crypt depth of the jejunal mucosa were determined by a single experimenter, blind to the pig breed and source of gut microbiota, and measured at 40 × magnification with an Olympus CK 40 microscope (Olympus Optical Company).
Antioxidant capacity and the digestive and absorptive enzyme activities
The jejunal crude enzyme solution was prepared according to procedure previously described by Diao et al. (2015) . The jejunum were collected and homogenized in 10 mmol/L ice-cold phosphate-buffered saline (m:v = 1:9). Homogenates were centrifuged at 2500 rpm at 4 °C for 10 min, and then the supernatant was used for subsequent analysis of the antioxidant capacity (methane dicarboxylic aldehyde (MDA), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC)), and digestive and absorptive enzyme activities (protein content, amylase, lipase, trypsin, lactase, sucrase, maltase, alkaline phosphatase (AKP), Na+, K+-ATPase, Ca+, Mg+-ATPase and γ-glutamyltransferase (γ-GT)), which were measured by the commercial kits (Nanjing Jiancheng Institute of Bioengineering, Jiangsu, China) with UV–VIS Spectrophotometer (UV1100, MAPADA, Shanghai, China) according to the manufacturers’ protocols.
Total RNA extraction, reverse transcription reaction and real-time quantitative PCR
Total RNA was isolated from the frozen duodenum, jejunum and ileum using the TRIzol reagent (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) according to the manufacturer’s protocol. The synthesis of the first strand of cDNA of each sample was obtained by reverse transcription by RT Reagents (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China) according to the manufacturer’s instructions. The genes which were relative to cell differentiation (CDX2, caudal-related homeodomain transcription 2), intestinal development (EGF, epidermal growth factor; GLP-2, glucagon-like peptide-2; ANG4, angiogenin 4; IGF-1, insulin-like growth factor-1; IGF-1R, insulin-like growth factor-1 receptor), digestion and absorption (SGLT-1, sodium/glucose cotransporter 1; GLUT-2, glucose transporter type 2; ZNT1, zinc transporters-1; DMT1, divalent metal transporter-1; SLC7A1,solute carrier family 7), and intestinal barrier (MUC1, mucin 1; MUC2, mucin 2; REGIIIγ, regeneration protein IIIγ; Occludin; ZO-1, zonula occludens 1) can be detected by real-time quantitative PCR with the CFX96 Real-Time PCR Detection System (Bio-Rad Laboratories, Richmond, CA) as described by Zhao et al. (2014) . The primers were synthesized commercially by Life Technologies Limited, which were listed in Additional file 1: Table S1.
Data were analyzed by ANONA using the statistical program SAS 9.2 (SAS Inst. Inc., NC) where each pig or mouse was the statistical unit. All differences were considered significant at P < 0.05 and were considered a trend at 0.05 ≤ P < 0.10. Principal coordinates analysis (PCoA) plots were produced using weighted UniFrac metrics. Plots were visualized using the R software (Packages ape). Log10-transformation was applied on the genus relative abundance data matrix for the heatmaps representation, which allowed visualizing similarities or differences between samples.
16S rRNA analysis of bacterial communities
The three groups of pig donors and their mice recipients exhibited different intestinal index and morphology
Intestinal index and morphology
Effect of genotype on intestinal index and morphology in pigs
Relative length of SI (mm/g)
Relative length of LI (mm/g)
Relative length of I (mm/g)
Relative density of SI (g/cm)
Relative density of LI (g/cm)
Relative density of I (g/cm)
Relative weight of SI (%)
Relative weight of LI (%)
Relative weight of I (%)
Villus height (μm)
Crypt depth (μm)
Villus height: crypt depth
Villus height (μm)
Crypt depth (μm)
Villus height: crypt depth
Villus height (μm)
Crypt depth (μm)
Villus height: crypt depth
Effect of gut flora source on intestinal index and morphology in mice
Relative length of SI (cm/g)
Relative length of LI (cm/g)
Relative length of I (cm/g)
Relative density of SI (g/cm)
Relative density of LI (g/cm)
Relative density of I (g/cm)
Relative weight of SI (%)
Relative weight of LI (%)
Relative weight of I (%)
Villus height (μm)
Crypt depth (μm)
Villus height: crypt depth
Villus height, μm
Crypt depth, μm
Villus height: crypt depth
Villus height (μm)
Crypt depth (μm)
Villus height: crypt depth
The villus height of duodenum and jejunum in TP was increased compared with YP and RP, and the villus height of ileum in TP and YP was higher than that in RP (P < 0.05) (Table 1). TP had higher crypt depth of duodenum than RP, YP had higher crypt depth of jejunum than RP, and TP and YP had higher crypt depth of ileum than RP (P < 0.05). The villus height: crypt depth of jejunum in RP was higher than that in YP (P < 0.05). Similarly, the villus height of duodenum and jejunum in TFM was higher than that in other two groups (P < 0.05) (Table 2). Moreover, there was a tendency towards increased jejunal crypt depth in TFM compared to YFM and RFM (P = 0.059).
Intestinal cell proliferation and differentiation
The relative mRNA expression of intestinal development-related genes
The digestive and absorptive ability was different among the three groups of pig donors and their mice recipients
Digestive and absorptive enzyme activity in jejunum
Effect of genotype on digestive and absorptive enzyme activity in jejunum of pigs
Total protein (mgprot/ml)
Na+, K+-ATPase (μmolPi/mgprot/hour)
Ca+, Mg+-ATPase (μmolPi/mgprot/hour)
Effect of gut flora source on digestive and absorptive enzyme activities in jejunum of mice
Na+, K+-ATPase (μmolPi/mgprot/hour)
Ca+, Mg+-ATPase (μmolPi/mgprot/hour)
Effect of genotype on jejunal antioxidant capacity in pigs
Digestion and absorption-related genes in small intestine
Effect of gut flora source on jejunal antioxidant capacity in mice
Tibetan pigs derived microbiota improved gut barrier in mice recipients
Jejunal antioxidant capacity
The concentration of MDA in YP was higher than that in TP and RP (P = 0.078) (Table 5). Likewise, compared with TFM and RFM, YFM exhibited increased MDA concentration in jejunum (P = 0.098) (Table 6). In addition, TFM had higher T-AOC activity compared with YFM and RFM (P = 0.073).
The relative mRNA expression of intestinal barrier-related genes
Gut microbiota has been recognized as a strong determinant factor of host physiology, especially its critical role in host gut development . The causal relationship between gut microbiota and host phenotypes has been widely studied via fecal microbiota transplantation. It has been shown that inflammatory bowel disease could be cured by transplanting fecal microbota from healthy human . Previous studies indicated that the gut of individuals and animals with different genotype or phenotype may harbor distinct microbiota [16, 17, 37]. However, there is not enough evidence to indicate the relationship between gut microbiota and gut phenotype features. Here, we demonstrated that different pig breeds exhibit a distinct gut microbial profile. Moreover, we found that transfering porcine microbiota to mice recipients can replicate the gut characteristics of the pig donors.
16S rRNA gene sequencing in donor pigs and recipient mice showed that the fecal microbiota composition at the phylum and genus levels were widely different in TP, YP and RP, and so were the same phenomena in TFM, YFM and RFM. More specifically, lower Firmicutes and higher Bacteroidetes levels were observed in YP and YFM, whereas higher levels of bacteria from Elusimicrobia, Fibrobacteres and Spirochaetes were observed in TP, and a higher level of Spirochaetes was observed in TFM. Studies comparing the gut microbiota between obese and lean animals showed that lower Firmicutes and higher Bacteroidetes levels were associated with lean phenotype [18, 38]. As is known to us, YP is an imported breed and is characterized by high body lean mass, which is consistent with our study. Spirochaetes is capable of degrading polymers, such as xylan, pectin and arabinogalactan, and is found to be positively correlated with the apparent hemicellulose digestibility of pigs [39–41]. Elusimicrobia is the intracellular symbiont of termite gut flagellates, Fibrobacteres is an important phylum of cellulose-degrading bacteria, and both of them are capable of degrading fiber [42, 43]. TP were recognized to be more adaptable to poor dietary conditions than foreign pig breeds , and this may be attributed to higher spirochaetes, Fibrobacteres and Elusimicrobia proportion in gut microbiota. In addition, for the genus level, a higher proportion of Prevotella and a lower proportion of Ruminococcus were found in healthy weight adolescents compared with obese humans [44, 45], which is similar to our study, a higher proportion of Prevotella and lower proportion of Ruminococcus were found in YP and YFM. Roseburia and Blautia are major bacteria that produce butyrate and acetic acid respectively [46, 47]. In our study, TP and TFM exhibited increased abundance of Roseburia and Blautia compared with YP and YFM, which is beneficial to gut health. Moreover, Lactobacillus and Parabacteroides are found to cure enteritis [48, 49]. In the present study, TP and TFM had higher proportion of Lactobacillus and Parabacteroides, and this may be attributed to better gut characteristics in TP.
Previous studies indicated that gut communities in the same phenotype individuals were similar to each other [37, 50]. We also found that TP, YP and RP fecal microbiota could be divided into three separate clusters based on PCA through 16S rRNA gene sequencing. Previous studies have shown that human and rat microbiota can be transferred to GF mice with striking preservation of structure and diversity [51, 52], which was consistent with our study, recipient mice exhibited a high similarity in bacterial community structure with their corresponding pig donors. From the above, gut composition differs between TP, YP and RP, and the mice recipients share high similarity in the gut microbiota with their pig donors.
Long-term reproductive and environmental isolation may lead to specific profiles of organ development, digestive ability and nutrient deposition existing in different pig breeds during their adaptation and evolution [53, 54]. In the present study, the relative weight of total intestine in Chinese indigenous pig breeds (TP and RP) and their mice recipients (TFM and RFM) was significantly higher than that in YP and YFM, which was in agreement with the previous finding that indicated the lean-type pigs had a lower proportional weight of intestine than indigenous genotypes . But many findings elucidated that pigs with leaner carcasses exhibited higher weights for small intestine and large intestine [54, 56]. An experiment with growth stages has shown that the development changes of relative viscera weight between lean pigs and fatter pigs was dissimilar in different growth stage [57, 58]. Thus, further research is clearly warranted. In this study, the intestine length did not differ between RP and YP, and the results were in accordance with previous findings , which indicated that the length of small intestine did not differ between foreign pig breeds and native pig breeds. However, the intestine length was higher in TP and TFM compared with YP and YFM in the current experiment. The differences of gut microbial ecology were observed between pig breeds, and changes in microbiota composition would affect the endogenous intestinotrophic proglucagon-derived peptide (GLP-2) production [38, 59]. It is well-known that IGF-1, EGF, GLP-2, and its receptor (GLP-2R) are important regulators of intestine length [60, 61], and ANG4 is a paneth cell granule protein shaping intestinal angiogenesis . In this study, TP and TFM had highest GLP-2 mRNA expression among the three pig breeds and their mice recipients, and TFM had higher ANG4 mRNA expression compared with YFM, which are associated with higher intestine length in TP and TFM.
Previous reports confirmed that pig breeds produce variation in the structure of the small intestine . In the present study, variations in small intestinal morphology were also observed among pig breeds and their mice recipients. Higher villus height of duodenum and jejunum was observed in TP and TFM compared with YP and RP, YFM and RFM, respectively. This is in accordance with data previously reported, villus surface was larger for Iberian pigs than Landrace × Large White pigs at 15 kg of BW . As we know, the CDX2 and AKP are involved in intestinal cell proliferation and differentiation, which could contribute to the structure of the intestine [64, 65]. In our study, we found that TP displayed higher CDX2 mRNA expression in the ileum and AKP activity in the jejunum compared with YP, and TFM exhibited higher CDX2 mRNA expression in the ileum compared with YFM. These results are generally consistent with that of Rubio et al. (2010) who reported differences in AKP activity were found among pig breeds . The abovementioned positive regulators must have a potent nutritional effect on the intestine growth and development, and these factors are differently expressed in the digestive tract among pig breeds and their mice recipients to account for differences in phenotype.
Small intestinal nutrient digestion and absorption was also affected by pig breeds. It has been observed that pig genotype (Iberian v. Landrace × Large White) exhibited different nitrogen retention and apparent total tract digestibility of nitrogen, energy and organic matter . In a study of changes in small intestinal nutrient transport in Meishan pigs and Yorkshire pigs showed that different glucose, arginine, glutamine and threonine transportation in the small intestine were observed in the two pig breeds . The ratio of bacteroidetes and firmicutes bacterial groups in the gut was increased in lean-type pigs which exhibited greater ability to absorb nutrients . Consistent with previous work , which observed that Landrace × Large White pigs had higher activities of lactase, sucrase and maltase than Iberian pigs in the small intestine, our current study showed YP and YFM had elevated jejunal sucrase and maltase activities, which may have resulted from its higher ratio of bacteroidetes and firmicutes bacterial groups. Meanwhile, TP had higher jejunal amylase, and trypsin than YP or RP, which is generally consistent with the previous study of two pig breeds, total activities of lipase, trypsin and amylase at 49 d of age were 2.0, 1.5 and 5.0 fold higher, respectively, in Alentejano piglets than Large White piglets . In our study, TP and TFM had higher jejunal γ-GT and relative SGLT1 mRNA expression, suggesting the differences of absorptive enzymes and transport carriers possibly affect nutrient digestion and absorption in different pig breeds. In addition, YP and YFM had highest MDA concentration in the jejunum among the three pig genotypes and their mice recipients in this study, and there is currently no available information on the difference in intestinal antioxidant capacity among pig breeds, and thus no comparisons could be made with other studies. Nevertheless, there have been some studies regarding meat antioxidant capacity among pig breeds, which exhibited differences of catalase and SOD in muscle among Pietrain, Landrace, Large-White, lberian, and lberian-Duroc . Taken together, the digestion and absorption function in the three pig genotypes are distinct, which is accompanied by different digestive and absorptive enzymes activities, transport carriers abundance and antioxidant capacity in the intestine.
Pig breeds also differ for their intestinal barrier. Previous study has shown that Yorkshire pigs responded to LPS by increasing resistance (decreasing conductance), whereas resistance in Meishan intestine did not change with LPS, which indicated that intestinal barrier function in traditional pig breed was better than Yorkshire pigs . Innate defenses, such as epithelial production of a-defensins and mucins, help prevent bacteria from crossing the mucosal barrier . ZO-1 is a junctional adaptor protein that interacts with multiple other junctional components, including the transmembrane proteins of the claudin, occludin and JAM families, which is important for intestinal integrity and barrier function . Furthermore, the direct host-commensal interaction is dictated by the presence of the intestinal mucus layer, and the expression and activation of mucin and ZO-1 are induced by exposing to a developing gut microbial community or their structural components, as well as the presence of products of bacterial metabolism . In our study, the relative ZO-1 mRNA expression of the ileum in TP and TFM was higher than that in YP and YFM, and RP and RFM had the highest relative MUC2 mRNA expression in the duodenum, suggesting native pig breeds (TP and RP) had better gut barrier function than foreign pig breeds (YP), which verified their stronger adaptability and disease resistance. As mentioned above, the differences in intestine characteristics including intestinal development and gut barrier among pig breeds could transmit to their recipients by fecal microbiota transplantation, and the intestinal microorganisms are essential for intestine development of mammals.
Our study showed that the microbiota affected gut characteristics through their impacts on epithelial renewal rate, morphology, nutrient digestion and absorption, and the intestinal barrier. Thus, while bacteria in the gut are highly variable, the influence of the microbiota in the intestine has far-reaching effects on host physiology. Short chain fatty acids (SCFA) were generated by bacterial fermentation of dietary polysaccharides, and it significantly affects energy metabolism, intestine morphology and immune function of the host via activation of its receptor GPR41 and GPR43 [71, 72]. Moreover, gut microbiota play a key role in determining bile acid (BA) profiles and subsequent effects on host gene expression . BA was reported to differentially activate BA receptors, which function as systemic signaling molecules to regulate host metabolism . Gut microbiota affect pig characteristics and transfer these phenotypes to recipient mice via aforementioned mechanisms in the current study. This study provides new approaches to intervene in animal production and health.
This study indicates that the microbiota composition differed among pig breeds, and TP had better intestinal morphology and barrier function than YP. The differences in gut-phenotypes among pig breeds would partially convey to recipient mice by fecal microbiota transplantation, especially intestinal morphology and enzyme activities. It is concluded that intestinal microbiota could transfer host gut characteristics from pigs to mice.
The authors wish to thank YH Luo and ZQ Huang from Institute of Animal Nutrition, Sichuan Agricultural University, helped with English language checking and editing of our manuscript. We thank the veterinarians XP Shi from College of Veterinary Medicine, Sichuan Agricultural University, for the elaborative animal examination and sampling. We thank Kevin Morgen who is a native English speaker for the modification of the manuscript. Thanks also go to JQ Li and XH Jiang from the Shenzhen Jincheng Laboratory Animal Technologies Co., LTD. for their excellent technical assistance and germ-free mice feeding.
This study was financially supported by the earmarked fund for the China Agriculture Research System (CARS-36). Sichuan Province Science and Technology Support Project (2012NZ0001), the National Natural Science Foundation of China (81370906), the National Basic Research Program of China (2013CB531406), and the National High Technology Research and Development Program of China (2014AA022209).
Availability of supporting data
16S rRNA gene sequences supporting the results of this article are available in the GenBank Database (https://0-www-ncbi-nlm-nih-gov.brum.beds.ac.uk/genbank/) under the accession number SRP078964.
DWC, HW and XBM conceived the study and designed the experiment. HD, HLY and YX performed the experiments, including chemical analysis, analyzed the experimental data, and wrote the manuscript. JY and BY verified the validity of the experimentation and checked the results. JH, PZ and BHZ participated in its design and gave important intellectual advice for approval. All of the authors read and approved the final version of this manuscript.
The authors declare that they have no competing interests.
Consent for publication
Ethics approval and consent to participate
All experimental procedures and animal care were accomplished in accordance with the Guide for the Care and Use of Laboratory Animals provided by the Institutional Animal Care Advisory Committee for Sichuan Agricultural University. All animal protocols used in this study were approved by the Animal Care and Use Committee of Sichuan Agricultural University under permit number DKY-B20131704.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
- Sekirov I, Russell SL, Antunes LC, Finlay BB. Gut microbiota in health and disease. Physiol Rev. 2010;90:859–904. doi:10.1152/physrev.00045.2009.View ArticlePubMedGoogle Scholar
- Ley RE, Lozupone CA, Hamady M, Knight R, Gordon JI. Worlds within worlds: evolution of the vertebrate gut microbiota. Nat Rev Microbiol. 2008;6:776–88.View ArticlePubMedPubMed CentralGoogle Scholar
- Hufeldt MR, Nielsen DS, Vogensen FK, Midtvedt T, Hansen AK. Variation in the gut microbiota of laboratory mice is related to both genetic and environmental factors. Comp Med. 2010;60:336–47.PubMedPubMed CentralGoogle Scholar
- De Filippo C, Cavalieri D, Di Paola M, Ramazzotti M, Poullet JB, Massart S, Collini S, Pieraccini G, Lionetti P. Impact of diet in shaping gut microbiota revealed by a comparative study in children from Europe and rural Africa. Proc Natl Acad Sci. 2010;107:14691–6.View ArticlePubMedPubMed CentralGoogle Scholar
- Spor A, Koren O, Ley R. Unravelling the effects of the environment and host genotype on the gut microbiome. Nat Rev Microbiol. 2011;9:279–90.View ArticlePubMedGoogle Scholar
- Benson AK, Kelly SA, Legge R, Ma F, Low SJ, Kim J, Zhang M, Oh PL, Nehrenberg D, Hua K. Individuality in gut microbiota composition is a complex polygenic trait shaped by multiple environmental and host genetic factors. Proc Natl Acad Sci. 2010;107:18933–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Pajarillo EAB, Chae JP, Balolong MP, Kim HB, Seo K-S, Kang D-K. Pyrosequencing-based analysis of fecal microbial communities in three purebred pig lines. J Microbiol. 2014;52:646–51.View ArticlePubMedGoogle Scholar
- Smith MI, Yatsunenko T, Manary MJ, Trehan I, Mkakosya R, Cheng J, Kau AL, Rich SS, Concannon P, Mychaleckyj JC, Liu J, Houpt E, Li JV, Holmes E, Nicholson J, Knights D, Ursell LK, Knight R, Gordon JI. Gut microbiomes of Malawian twin pairs discordant for kwashiorkor. Science. 2013;339:548–54. doi:10.1126/science.1229000.View ArticlePubMedPubMed CentralGoogle Scholar
- Turnbaugh PJ, Hamady M, Yatsunenko T, Cantarel BL, Duncan A, Ley RE, Sogin ML, Jones WJ, Roe BA, Affourtit JP, Egholm M, Henrissat B, Heath AC, Knight R, Gordon JI. A core gut microbiome in obese and lean twins. Nature. 2009;457:480–4. doi:10.1038/nature07540.View ArticlePubMedGoogle Scholar
- Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Microbial ecology: human gut microbes associated with obesity. Nature. 2006;444:1022–3.View ArticlePubMedGoogle Scholar
- Vijay-Kumar M, Aitken JD, Carvalho FA, Cullender TC, Mwangi S, Srinivasan S, Sitaraman SV, Knight R, Ley RE, Gewirtz AT. Metabolic syndrome and altered gut microbiota in mice lacking Toll-like receptor 5. Science. 2010;328:228–31.View ArticlePubMedPubMed CentralGoogle Scholar
- Yang S, Zhang H, Mao H, Yan D, Lu S, Lian L, Zhao G, Yan Y, Deng W, Shi X, Han S, Li S, Wang X, Gou X. The local origin of the Tibetan pig and additional insights into the origin of Asian pigs. PLoS One. 2011;6:e28215. doi:10.1371/journal.pone.0028215.View ArticlePubMedPubMed CentralGoogle Scholar
- Zeng Z, Yu B, Mao X, Chen D. Effects of dietary digestible energy concentration on growth, meat quality, and PPARγ gene expression in muscle and adipose tissues of Rongchang piglets. Meat Sci. 2012;90:66–70.View ArticlePubMedGoogle Scholar
- Knapp P, Willam A, Sölkner J. Genetic parameters for lean meat content and meat quality traits in different pig breeds. Livest Prod Sci. 1997;52:69–73.View ArticleGoogle Scholar
- Renaudeau D, Mourot J. A comparison of carcass and meat quality characteristics of Creole and Large White pigs slaughtered at 90 kg BW. Meat Sci. 2007;76:165–71.View ArticlePubMedGoogle Scholar
- Luo YH, Su Y, Wright AD, Zhang LL, Smidt H, Zhu WY. Lean breed Landrace pigs harbor fecal methanogens at higher diversity and density than obese breed Erhualian pigs. Archaea. 2012;2012:605289. doi:10.1155/2012/605289.View ArticlePubMedPubMed CentralGoogle Scholar
- Yamano H, Koike S, Kobayashi Y, Hata H. Phylogenetic analysis of hindgut microbiota in Hokkaido native horses compared to light horses. Anim Sci J. 2008;79:234–42.View ArticleGoogle Scholar
- Guo X, Xia X, Tang R, Zhou J, Zhao H, Wang K. Development of a real-time PCR method for Firmicutes and Bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs. Lett Appl Microbiol. 2008;47:367–73. doi:10.1111/j.1472-765X.2008.02408.x.View ArticlePubMedGoogle Scholar
- Shanahan F. The host-microbe interface within the gut. Best Pract Res Clin Gastroenterol. 2002;16:915–31.View ArticlePubMedGoogle Scholar
- Banasaz M, Norin E, Holma R, Midtvedt T. Increased enterocyte production in gnotobiotic rats mono-associated with Lactobacillus rhamnosus GG. Appl Environ Microbiol. 2002;68:3031–4.View ArticlePubMedPubMed CentralGoogle Scholar
- Gordon HA, Bruckner-Kardoss E. Effect of normal microbial flora on intestinal surface area. Am J Physiol. 1961;201:175–8.PubMedGoogle Scholar
- O'Hara AM, Shanahan F. The gut flora as a forgotten organ. EMBO Rep. 2006;7:688–93. doi:10.1038/sj.embor.7400731.View ArticlePubMedPubMed CentralGoogle Scholar
- Wostmann B, Bruckner-Kardoss E. Development of cecal distention in germ-free baby rats. Am J Physiol. 1959;197:1345–6.PubMedGoogle Scholar
- Pamer EG. Fecal microbiota transplantation: effectiveness, complexities, and lingering concerns. Mucosal Immunol. 2014;7:210–4. doi:10.1038/mi.2013.117.View ArticlePubMedGoogle Scholar
- Tlaskalova-Hogenova H, Stepankova R, Kozakova H, Hudcovic T, Vannucci L, Tuckova L, Rossmann P, Hrncir T, Kverka M, Zakostelska Z, Klimesova K, Pribylova J, Bartova J, Sanchez D, Fundova P, Borovska D, Srutkova D, Zidek Z, Schwarzer M, Drastich P, Funda DP. The role of gut microbiota (commensal bacteria) and the mucosal barrier in the pathogenesis of inflammatory and autoimmune diseases and cancer: contribution of germ-free and gnotobiotic animal models of human diseases. Cell Mol Immunol. 2011;8:110–20. doi:10.1038/cmi.2010.67.View ArticlePubMedPubMed CentralGoogle Scholar
- Hapfelmeier S, Lawson MA, Slack E, Kirundi JK, Stoel M, Heikenwalder M, Cahenzli J, Velykoredko Y, Balmer ML, Endt K, Geuking MB, Curtiss 3rd R, McCoy KD, Macpherson AJ. Reversible microbial colonization of germ-free mice reveals the dynamics of IgA immune responses. Science. 2010;328:1705–9. doi:10.1126/science.1188454.View ArticlePubMedPubMed CentralGoogle Scholar
- Holmes E, Li JV, Marchesi JR, Nicholson JK. Gut microbiota composition and activity in relation to host metabolic phenotype and disease risk. Cell Metab. 2012;16:559–64. doi:10.1016/j.cmet.2012.10.007.View ArticlePubMedGoogle Scholar
- Khoruts A, Sadowsky MJ. Therapeutic transplantation of the distal gut microbiota. Mucosal Immunol. 2011;4:4–7. doi:10.1038/mi.2010.79.View ArticlePubMedGoogle Scholar
- Aroniadis OC, Brandt LJ. Fecal microbiota transplantation: past, present and future. Curr Opin Gastroenterol. 2013;29:79–84.View ArticlePubMedGoogle Scholar
- HuiYu Z (2008) The charaeteristies of growth of Tibetan pig in Sichuan Ganzi Sichuan Agrieultural UniversityGoogle Scholar
- Hamilton MJ, Weingarden AR, Sadowsky MJ, Khoruts A. Standardized frozen preparation for transplantation of fecal microbiota for recurrent Clostridium difficile infection. Am J Gastroenterol. 2012;107:761–7. doi:10.1038/ajg.2011.482.View ArticlePubMedGoogle Scholar
- Zeng B, Li G, Yuan J, Li W, Tang H, Wei H. Effects of age and strain on the microbiota colonization in an infant human flora-associated mouse model. Curr Microbiol. 2013;67:313–21. doi:10.1007/s00284-013-0360-3.View ArticlePubMedGoogle Scholar
- Bruce-Keller AJ, Salbaum JM, Luo M, Blanchard E, Taylor CM, Welsh DA, Berthoud H-R. Obese-type gut microbiota induce neurobehavioral changes in the absence of obesity. Biol Psychiatry. 2015;77:607–15.View ArticlePubMedGoogle Scholar
- Mao X, Gu C, Hu H, Tang J, Chen D, Yu B, He J, Yu J, Luo J, Tian G. Dietary Lactobacillus rhamnosus GG Supplementation Improves the Mucosal Barrier Function in the Intestine of Weaned Piglets Challenged by Porcine Rotavirus. PLoS One. 2016;11:e0146312. doi:10.1371/journal.pone.0146312.View ArticlePubMedPubMed CentralGoogle Scholar
- Diao H, Zheng P, Yu B, He J, Mao X, Yu J, Chen D. Effects of benzoic Acid and thymol on growth performance and gut characteristics of weaned piglets. Asian-Australas J Anim Sci. 2015;28:827–39. doi:10.5713/ajas.14.0704.View ArticlePubMedPubMed CentralGoogle Scholar
- Zhao Y, Yu B, Mao X, He J, Huang Z, Zheng P, Yu J, Han G, Liang X, Chen D. Dietary vitamin D supplementation attenuates immune responses of pigs challenged with rotavirus potentially through the retinoic acid-inducible gene I signalling pathway. Br J Nutr. 2014;112:381–9. doi:10.1017/S000711451400097X.View ArticlePubMedGoogle Scholar
- Turnbaugh PJ, Backhed F, Fulton L, Gordon JI. Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe. 2008;3:213–23. doi:10.1016/j.chom.2008.02.015.View ArticlePubMedPubMed CentralGoogle Scholar
- Guo X, Xia X, Tang R, Wang K. Real-time PCR quantification of the predominant bacterial divisions in the distal gut of Meishan and Landrace pigs. Anaerobe. 2008;14:224–8. doi:10.1016/j.anaerobe.2008.04.001.View ArticlePubMedGoogle Scholar
- Lilburn T, Schmidt T, Breznak J. Phylogenetic diversity of termite gut spirochaetes. Environ Microbiol. 1999;1:331–45.View ArticlePubMedGoogle Scholar
- Paster B, Canale-Parola E. Physiological diversity of rumen spirochetes. Appl Environ Microbiol. 1982;43:686–93.PubMedPubMed CentralGoogle Scholar
- Niu Q, Li P, Hao S, Zhang Y, Kim SW, Li H, Ma X, Gao S, He L, Wu W, Huang X, Hua J, Zhou B, Huang R. Dynamic distribution of the gut microbiota and the relationship with apparent crude fiber digestibility and growth stages in pigs. Sci Rep. 2015;5:9938. doi:10.1038/srep09938.View ArticlePubMedPubMed CentralGoogle Scholar
- Ransom-Jones E, Jones DL, McCarthy AJ, McDonald JE. The Fibrobacteres: an important phylum of cellulose-degrading bacteria. Microb Ecol. 2012;63:267–81. doi:10.1007/s00248-011-9998-1.View ArticlePubMedGoogle Scholar
- Ikeda-Ohtsubo W, Faivre N, Brune A. Putatively free-living 'Endomicrobia'- ancestors of the intracellular symbionts of termite gut flagellates? Environ Microbiol Rep. 2010;2:554–9. doi:10.1111/j.1758-2229.2009.00124.x.View ArticlePubMedGoogle Scholar
- Santacruz A, Marcos A, Wärnberg J, Martí A, Martin‐Matillas M, Campoy C, Moreno LA, Veiga O, Redondo‐Figuero C, Garagorri JM. Interplay between weight loss and gut microbiota composition in overweight adolescents. Obesity. 2009;17:1906–15.View ArticlePubMedGoogle Scholar
- Ferrer M, Ruiz A, Lanza F, Haange SB, Oberbach A, Till H, Bargiela R, Campoy C, Segura MT, Richter M, von Bergen M, Seifert J, Suarez A. Microbiota from the distal guts of lean and obese adolescents exhibit partial functional redundancy besides clear differences in community structure. Environ Microbiol. 2013;15:211–26. doi:10.1111/j.1462-2920.2012.02845.x.View ArticlePubMedGoogle Scholar
- Duncan SH, Aminov RI, Scott KP, Louis P, Stanton TB, Flint HJ. Proposal of Roseburia faecis sp. nov., Roseburia hominis sp. nov. and Roseburia inulinivorans sp. nov., based on isolates from human faeces. Int J Syst Evol Microbiol. 2006;56:2437–41.View ArticlePubMedGoogle Scholar
- Liu C, Finegold SM, Song Y, Lawson PA. Reclassification of Clostridium coccoides, Ruminococcus hansenii, Ruminococcus hydrogenotrophicus, Ruminococcus luti, Ruminococcus productus and Ruminococcus schinkii as Blautia coccoides gen. nov., comb. nov., Blautia hansenii comb. nov., Blautia hydrogenotrophica comb. nov., Blautia luti comb. nov., Blautia producta comb. nov., Blautia schinkii comb. nov. and description of Blautia wexlerae sp. nov., isolated from human faeces. Int J Syst Evol Microbiol. 2008;58:1896–902.View ArticlePubMedGoogle Scholar
- Kverka M, Zakostelska Z, Klimesova K, Sokol D, Hudcovic T, Hrncir T, Rossmann P, Mrazek J, Kopecny J, Verdu EF, Tlaskalova-Hogenova H. Oral administration of Parabacteroides distasonis antigens attenuates experimental murine colitis through modulation of immunity and microbiota composition. Clin Exp Immunol. 2011;163:250–9. doi:10.1111/j.1365-2249.2010.04286.x.View ArticlePubMedPubMed CentralGoogle Scholar
- Partty A, Lehtonen L, Kalliomaki M, Salminen S, Isolauri E. Probiotic Lactobacillus rhamnosus GG therapy and microbiological programming in infantile colic: a randomized, controlled trial. Pediatr Res. 2015;78:470–5. doi:10.1038/pr.2015.127.View ArticlePubMedGoogle Scholar
- Ley RE, Bäckhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI. Obesity alters gut microbial ecology. Proc Natl Acad Sci U S A. 2005;102:11070–5.View ArticlePubMedPubMed CentralGoogle Scholar
- Duca FA, Sakar Y, Lepage P, Devime F, Langelier B, Doré J and Covasa M (2014) Replication of obesity and associated signaling pathways through transfer of microbiota from obese prone rat. Diabetes:DB_131526.Google Scholar
- Turnbaugh PJ, Ridaura VK, Faith JJ, Rey FE, Knight R, Gordon JI. The effect of diet on the human gut microbiome: a metagenomic analysis in humanized gnotobiotic mice. Sci Transl Med. 2009;1:6ra14.View ArticlePubMedPubMed CentralGoogle Scholar
- Albin DM, Wubben JE, Rowlett JM, Tappenden KA, Nowak RA. Changes in small intestinal nutrient transport and barrier function after lipopolysaccharide exposure in two pig breeds. J Anim Sci. 2007;85:2517–23. doi:10.2527/jas.2006-237.View ArticlePubMedGoogle Scholar
- Jiang YZ, Zhu L, Li XW, Si T. Evaluation of the Chinese indigenous pig breed Dahe and crossbred Dawu for growth and carcass characteristics, organ weight, meat quality and intramuscular fatty acid and amino acid composition. Animal. 2011;5:1485–92. doi:10.1017/S1751731111000425.View ArticlePubMedGoogle Scholar
- Noblet J, Karege C, Dubois S, van Milgen J. Metabolic utilization of energy and maintenance requirements in growing pigs: effects of sex and genotype. J Anim Sci. 1999;77:1208–16.View ArticlePubMedGoogle Scholar
- Ruusunen M, Partanen K, Pösö R, Puolanne E. The effect of dietary protein supply on carcass composition, size of organs, muscle properties and meat quality of pigs. Livest Sci. 2007;107:170–81.View ArticleGoogle Scholar
- Barea R, Nieto R, Vitari F, Domeneghini C, Aguilera JF. Effects of pig genotype (Iberian v. Landrace x Large White) on nutrient digestibility, relative organ weight and small intestine structure at two stages of growth. Animal. 2011;5:547–57. doi:10.1017/S1751731110002181.View ArticlePubMedGoogle Scholar
- Rivera-Ferre MG, Aguilera JF, Nieto R. Muscle fractional protein synthesis is higher in Iberian than in Landrace growing pigs fed adequate or lysine-deficient diets. J Nutr. 2005;135:469–78.PubMedGoogle Scholar
- Cani PD, Possemiers S, Van de Wiele T, Guiot Y, Everard A, Rottier O, Geurts L, Naslain D, Neyrinck A, Lambert DM, Muccioli GG, Delzenne NM. Changes in gut microbiota control inflammation in obese mice through a mechanism involving GLP-2-driven improvement of gut permeability. Gut. 2009;58:1091–103. doi:10.1136/gut.2008.165886.View ArticlePubMedPubMed CentralGoogle Scholar
- Lovshin J, Yusta B, Iliopoulos I, Migirdicyan A, Dableh L, Brubaker PL, Drucker DJ. Ontogeny of the glucagon-like peptide-2 receptor axis in the developing rat intestine. Endocrinology. 2000;141:4194–201. doi:10.1210/endo.141.11.7773.PubMedGoogle Scholar
- Steeb CB, Trahair JF, Tomas FM, Read LC. Prolonged administration of IGF peptides enhances growth of gastrointestinal tissues in normal rats. Am J Physiol. 1994;266:G1090–8.PubMedGoogle Scholar
- Hooper LV. Bacterial contributions to mammalian gut development. Trends Microbiol. 2004;12:129–34. doi:10.1016/j.tim.2004.01.001.View ArticlePubMedGoogle Scholar
- Rubio LA, Ruiz R, Peinado MJ, Echavarri A. Morphology and enzymatic activity of the small intestinal mucosa of Iberian pigs as compared with a lean pig strain. J Anim Sci. 2010;88:3590–7. doi:10.2527/jas.2010-3040.View ArticlePubMedGoogle Scholar
- Wan X, Bi J, Gao X, Tian F, Wang X, Li N, Li J. Partial enteral nutrition preserves elements of Gut barrier function, including innate immunity, intestinal alkaline phosphatase (IAP) level, and intestinal microbiota in mice. Nutrients. 2015;7:6294–312. doi:10.3390/nu7085288.View ArticlePubMedPubMed CentralGoogle Scholar
- San Roman AK, Tovaglieri A, Breault DT, Shivdasani RA. Distinct processes and transcriptional targets underlie CDX2 requirements in intestinal stem cells and differentiated villus cells. Stem cell reports. 2015;5:673–81.View ArticlePubMedPubMed CentralGoogle Scholar
- Cui C, Shen CJ, Jia G, Wang KN. Effect of dietary Bacillus subtilis on proportion of Bacteroidetes and Firmicutes in swine intestine and lipid metabolism. Genet Mol Res. 2013;12:1766–76. doi:10.4238/2013.May.23.1.View ArticlePubMedGoogle Scholar
- Freire J, Peiniau J, Cunha L, Almeida J, Aumaitre A. Comparative effects of dietary fat and fibre in Alentejano and Large White piglets: digestibility, digestive enzymes and metabolic data. Livest Prod Sci. 1998;53:37–47.View ArticleGoogle Scholar
- Hernandez P, Zomeno L, Arino B, Blasco A. Antioxidant, lipolytic and proteolytic enzyme activities in pork meat from different genotypes. Meat Sci. 2004;66:525–9. doi:10.1016/S0309-1740(03)00155-4.View ArticlePubMedGoogle Scholar
- Hooper LV, Gordon JI. Commensal host-bacterial relationships in the gut. Science. 2001;292:1115–8.View ArticlePubMedGoogle Scholar
- Tornavaca O, Chia M, Dufton N, Almagro LO, Conway DE, Randi AM, Schwartz MA, Matter K, Balda MS. ZO-1 controls endothelial adherens junctions, cell-cell tension, angiogenesis, and barrier formation. J Cell Biol. 2015;208:821–38. doi:10.1083/jcb.201404140.View ArticlePubMedPubMed CentralGoogle Scholar
- Brown AJ, Goldsworthy SM, Barnes AA, Eilert MM, Tcheang L, Daniels D, Muir AI, Wigglesworth MJ, Kinghorn I, Fraser NJ, Pike NB, Strum JC, Steplewski KM, Murdock PR, Holder JC, Marshall FH, Szekeres PG, Wilson S, Ignar DM, Foord SM, Wise A, Dowell SJ. The Orphan G protein-coupled receptors GPR41 and GPR43 are activated by propionate and other short chain carboxylic acids. J Biol Chem. 2003;278:11312–9. doi:10.1074/jbc.M211609200.View ArticlePubMedGoogle Scholar
- Covington DK, Briscoe CA, Brown AJ, Jayawickreme CK. The G-protein-coupled receptor 40 family (GPR40-GPR43) and its role in nutrient sensing. Biochem Soc Trans. 2006;34:770–3. doi:10.1042/BST0340770.View ArticlePubMedGoogle Scholar
- Ridlon JM, Kang DJ, Hylemon PB. Bile salt biotransformations by human intestinal bacteria. J Lipid Res. 2006;47:241–59. doi:10.1194/jlr.R500013-JLR200.View ArticlePubMedGoogle Scholar
- Sayin SI, Wahlstrom A, Felin J, Jantti S, Marschall HU, Bamberg K, Angelin B, Hyotylainen T, Oresic M, Backhed F. Gut microbiota regulates bile acid metabolism by reducing the levels of tauro-beta-muricholic acid, a naturally occurring FXR antagonist. Cell Metab. 2013;17:225–35. doi:10.1016/j.cmet.2013.01.003.View ArticlePubMedGoogle Scholar