Optimization of Culture Medium for Pig Colonic Microbiome Prepared by Fermentation in Vitro

BAI Yu; YAN Qiao; LI Tiantian; ZHOU Xingjian; HE Beibei; TAO Shiyu; ZHANG Shuai; WANG Junjun

doi:10.3969/j.issn.1006-267x.2019.08.033

Chinese Journal of Animal Nutrition >

2019 , Vol. 31 >Issue 8: 3719 - 3726

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.08.033

Optimization of Culture Medium for Pig Colonic Microbiome Prepared by Fermentation in Vitro

BAI Yu ,
YAN Qiao ,
LI Tiantian ,
ZHOU Xingjian ,
HE Beibei ,
TAO Shiyu ,
ZHANG Shuai ,
WANG Junjun

Expand

State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received date: 2019-01-11

Online published: 2019-08-19

Fold

Abstract

The colonic microbial group of animals has important significance for the health of hindgut and nutrient utilization. However, there are many inconveniences in the study of intestinal microbial function and mechanism in vivo. Therefore, this study analyzed the nutrient composition of growing pig colonic digesta and VL culture medium, and then changed and optimized the nutritional composition of the VL culture medium to consistent with colonic digesta. simulated microbiome similar to the colonic microbiome was cultured by using the one-way continuous fermentation system in vitro, and the microbiota structural similarity between simulated microbiome and colonic microbiome was calculated. The results showed that the contents of crude protein and soluble sugar in colonic digesta was significantly different from those in VL culture medium (3.09% vs. 1.49%, 0.43% vs. 0.31%). In addition, after adjusting the crude protein and soluble sugar contents in VL culture medium to the levels of colonic digesta, and adding the additions of molasses (VLMOL), corn syrup dry powder (VLCSL), digestive tract secreted mucin (VLM) and their combination into the culture medium, the microbiota structural similarity between the colonic microbiome and the simulated microbiome cultured by VLMOL, VLCSL, VLM and ZH culture mediums was 68.5%, 52.3%, 10.7% and 5.0%, respectively. In term of total short chain fatty acid concentration, the total amount of volatile fatty acids produced by VLMOL cultured colonic microbial group was more similar with that of the colonic digesta (21.75 mmol/L), whereas that in fermentation broths produced by VLCSL, VLM and ZH culture mediums were significantly different compared with colonic digesta (43.95, 39.05, 42.68 mmol/L vs. 26.03 mmol/L). In conclusion, a highly reliable culture medium which optimized by adjusting the contents of crude protein and soluble sugar in VL culture medium based of these in colonic digesta and adding molasses into the modified VL culture medium, can be used to prepare the colonic microbiome of growing pig by fermentation in vitro.

Key words： pig; colon; microbiome; molasses; fermentation in vitro

Cite this article

BAI Yu , YAN Qiao , LI Tiantian , ZHOU Xingjian , HE Beibei , TAO Shiyu , ZHANG Shuai , WANG Junjun . Optimization of Culture Medium for Pig Colonic Microbiome Prepared by Fermentation in Vitro[J]. Chinese Journal of Animal Nutrition, 2019 , 31(8) : 3719 -3726 . DOI: 10.3969/j.issn.1006-267x.2019.08.033

References

[1] BÄCKHED F,LEY R E,SONNENBURG J L,et al.Host-bacterial mutualism in the human intestine[J].Science,2005,307(5717):1915-1920.

[2] QIN J J,LI R Q,RAES J,et al.A human gut microbial gene catalogue established by metagenomic sequencing[J].Nature,2010,464(7285):59-65.

[3] BAKKEN J S,BORODY T,BRANDT L J,et al.Treating Clostridium difficile infection with fecal microbiota transplantation[J].Clinical Gastroenterology and Hepatology,2011,9(12):1044-1049.

[4] GOUGH E,SHAIKH H,MANGES A R.Systematic review of intestinal microbiota transplantation (fecal bacteriotherapy) for recurrent Clostridium difficile infection[J].Clinical Infectious Diseases,2011,53(10):994-1002.

[5] 陈波,王宇,雷芳,等.肠道微生物体外模型研究进展[J].中国微生态学杂志,2012,24(8):766-768.
[6] YIN Y S,LEI F,ZHU L Y,et al.Exposure of different bacterial inocula to newborn chicken affects gut microbiota development and ileum gene expression[J].The ISME Journal,2010,4(3):367-376.

[7] SHEHATA A A,WIELAND S,ALDIN A A,et al.The effect of glyphosate on potential pathogens and beneficial members of poultry microbiota in vitro[J].Current Microbiology,2013,66(4):350-358.

[8] LEI F,YIN Y S,WANG Y Z,et al.Higher-level production of volatile fatty acids in vitro by chicken gut microbiotas than by human gut microbiotas as determined by functional analyses[J].Applied and Environmental Microbiology,2012,78(16):5763-5772.

[9] 丁雪梅, 张晓君, 赵云,等. 蒽酮比色法测定可溶性糖含量的试验方法改进[J]. 黑龙江畜牧兽医, 2016(12):230-233.
[10] ZHAO J B,LIU P,WU Y,et al.Dietary fiber increases butyrate-producing bacteria and improves the growth performance of weaned piglets[J].Journal of Agricultural and Food Chemistry,2018,66(30):7995-8004.

[11] 范彬,尹业师,孙刚,等.不同碳水化合物对正常人肠道菌群体外模拟效果的比较[J].中华内科杂志,2016,55(5):381-385.
[12] 李长青,金海,薛树媛.采用PCR-DGGE技术对放牧羊补饲糖蜜尿素舔砖后细菌种群结构的分析[J].黑龙江畜牧兽医,2015(10):112-115.
[13] COSTA A F S,ALMEIDA F C G,VINHAS G M,et al.Production of bacterial cellulose by Gluconacetobacter hansenii using corn steep liquor as nutrient sources[J].Frontiers in Microbiology,2017,8:2027.
[14] KNUDSEN K E B,JENSEN B B,HANSEN I.Digestion of polysaccharides and other major components in the small and large intestine of pigs fed on diets consisting of oat fractions rich in β-D-glucan[J].British Journal of Nutrition,1993,70(2):537-556.

[15] CARLSTEDT-DUKE B,MIDTVEDT T,NORD C E,et al.Isolation and characterization of a mucin-degrading strain of Peptostreptococcus from rat intestinal tract[J].Acta Pathologica,Microbiologica,et immunologica Scandinavica-Section B:Microbiology,1986,94(1/2/3/4/5/6):293-300.
[16] ROKHSEFAT S,LIN A F,COMELLI E M.Mucin-microbiota interaction during postnatal maturation of the intestinal ecosystem:clinical implications[J].Digestive Diseases and Sciences,2016,61(6):1473-1486.

[17] SIMON O.Protein digestion and absorption measured by re-entrant cannula in the pig[J].Nutrition Reviews,1979,37(5):149-150.
[18] KACZMAREK S A,BARRI A,HEJDYSZ M,et al.Effect of different doses of coated butyric acid on growth performance and energy utilization in broilers[J].Poultry Science,2016,95(4):851-859.

[19] BOSCH G,WRIGGLESWORTH D J,CONE J W,et al.Effects of preservation conditions of canine feces on in vitro gas production kinetics and fermentation end products[J].Journal of Animal Science,2013,91(1):259-267.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References