Effects of vitamin B complex on intestinal microflora composition and gut epithelial structure in growing goats

doi:10.11686/cyxb2020562

Abstract

Abstract:

The aim of this study was to explore the effects of vitamin B complex on intestinal microflora composition and intestinal morphology and structure in growing goats. Ten four-month-old female goats with similar body weight （12.60±1.28 kg） were randomly divided into a control group （CON） and a group treated with vitamin B complex （VB）. Goats in the CON group were fed a basal diet （n=5）， and goats in the VB group were fed with the basal diet supplemented with vitamin B complex （n=5）. The experiment comprised an adaptive feeding period of two weeks and a treatment period of 11 weeks. Throughout the experiment， all goats were provided free access to water. At the end of the experiment， the microflora composition of cecal and colonic contents was analyzed by 16S rRNA sequencing， and the intestinal epithelial morphology structure and expression levels of related genes and proteins expression were detected. Compared to CON， VB supplementation did not alter the α diversity index obtained by bacterial taxonomic analysis （P>0.05）. At the phylum level， Firmicutes and Bacteroidetes were the dominant bacteria in the intestines of the goats. Compared to CON， there was a tendency for the abundance of Bacteroidetes in the cecum of the VB group to increase （0.05<P<0.10） and for the abundance of Spirochaetes to decrease （0.05<P<0.10）. At the taxonomic order level， the abundance of Solirubrobacterales and Streptomycetales in the cecum of goats in the VB group was significantly decreased （P<0.05）. At the taxonomic family level， the abundance of Christensenellaceae， Streptomycetaceae， Solirubrobacteraceae in the cecum and Christensenellaceae in the colon of goats in the VB group was significantly decreased （P<0.05）. At the genus level， the abundance of Lachnoclostridium in the cecum of goats in the VB group was significantly increased （P<0.05）， while the abundance of Bacillus， Nocardioides in the cecum and unidentified_Christensenellaceae and unidentified_Gammaproteobacteria in the colon of goats in the VB group was significantly decreased （P<0.05）. Compared to CON， intestinal villus height （P<0.01）， crypt depth （P<0.05） and the ratio of villus height/crypt depth （V/C， P<0.05） of the jejunum in the VB group were significantly increased； and the intestinal crypt depth of the ileum was significantly decreased （P<0.05）. Real-time PCR results showed that Occludin （P<0.05）， EGFR （P<0.01）， MKI67 （P<0.05） and CCND （P<0.05） mRNA expression was significantly up-regulated in colonic epithelium of the VB group， and ZO-1 protein expression was also significantly up-regulated （P<0.05）. It is concluded that dietary supplemention with vitamin B complex increased the abundance of certain probiotics and reduce the abundance of harmful bacteria in the cecum and colon of growing goats. Dietary supplementation with VB also promoted the growth of intestinal villi and improved intestinal structure. The observed changes from VB supplementation would benefit animal welfare and growth.

Key words: vitamin B complex, intestinal microflora, gut epithelial structure, tight junction, goats

Chen WU, Zhi-hao YAO, Wen-qing MEI, Yu-yan FENG, Qu CHEN, Ying-dong NI. Effects of vitamin B complex on intestinal microflora composition and gut epithelial structure in growing goats[J]. Acta Prataculturae Sinica, 2021, 30(11): 170-180.

Figures/Tables 11

References 43

1	Beaudet V， Gervais R， Graulet B， et al. Effects of dietary nitrogen levels and carbohydrate sources on apparent ruminal synthesis of some B vitamins in dairy cows. Journal of Dairy Science， 2016， 99（4）： 2730-2739.
2	Hooper L V， Gordon J I. Commensal host-bacterial relationships in the gut. Science， 2001， 292（5519）： 1115-1118.
3	Rowland I， Gibson G， Heinken A， et al. Gut microbiota functions： Metabolism of nutrients and other food components. European Journal of Nutrition， 2018， 57（1）： 1-24.
4	Rastelli M， Cani P D， Knauf C. The gut microbiome influences host endocrine functions. Endocrine Reviews， 2019， 40（5）： 1271-1284.
5	Stefania D S， Elisabetta C， Mauro M， et al. Nutritional keys for intestinal barrier modulation. Frontiers in Immunology， 2015， 6： 612.
6	Long J H， Wang B W， Kong M， et al. Effects of vitamin B₁₂ supplemental level on growth performance， intestinal development and microflora structure in cecum of Wulong geese aged from 5 to 15 weeks. Chinese Journal of Animal Nutrition， 2018， 30（10）： 3930-3940.
	龙建华，王宝维，孔敏，等. 饲粮中维生素B₁₂添加水平对5~15周龄五龙鹅生长性能、肠道发育和盲肠菌群结构的影响. 动物营养学报， 2018， 30（10）： 3930-3940.
7	Wang B W， Wang X， Ge W H， et al. Effects of vitamin B₂ on growth performance， serum hormone contents and intestinal tissue structure of Wulong geese aged from 5 to 16 weeks. Chinese Journal of Animal Nutrition， 2014， 26（3）： 637-645.
	王宝维，王鑫，葛文华，等. 维生素B₂对5~16周龄五龙鹅生长性能、血清激素含量和肠道组织结构的影响. 动物营养学报， 2014， 26（3）： 637-645.
8	Halsted C H. The small intestine in vitamin B₁₂ and folate deficiency. Nutrition Reviews， 1975， 33（2）： 33-37.
9	Wang L， Shah A M， Liu Y， et al. Relationship between true digestibility of dietary phosphorus and gastrointestinal bacteria of goats. PLoS One， 2020， 15（5）： e0225018.
10	Jiang S， Huo D， You Z， et al. The distal intestinal microbiome of hybrids of Hainan black goats and Saanen goats. PLoS One， 2020， 15（1）： e0228496.
11	Clemmons B A， Voy B H， Myer P R. Altering the gut microbiome of cattle： Considerations of host-microbiome interactions for persistent microbiome manipulation. Microbial Ecology， 2019， 77（2）： 523-536.
12	Hoover W H. Digestion and absorption in the hindgut of ruminants. Journal of Animal Science， 1978， 46（6）： 1789-1799.
13	Ley R E， Hamady M， Lozupone C， et al. Evolution of mammals and their gut microbes. Science， 2008， 320（5883）： 1647-1651.
14	Seshadri R， Leahy S C， Attwood G T， et al. Cultivation and sequencing of rumen microbiome members from the Hungate1000 collection. Nature Biotechnology， 2018， 36（4）： 359-367.
15	Sasson G， Kruger Ben-shabat S， Seroussi E， et al. Heritable bovine rumen bacteria are phylogenetically related and correlated with the cow’s capacity to harvest energy from its feed. mBio， 2017， 8（4）： e00703-17.
16	Edwin E E， Hebert C N， Jackman R， et al. Thiamine requirement of young ruminants. Journal of Agricultural Science， 1976， 87（3）： 679-688.
17	Burkholder P R， Mcveigh I. Synthesis of vitamins by intestinal bacteria. Proceedings of the National Academy of Sciences of the United States of America， 1942， 28（7）： 285-289.
18	Yao Z H， Mei W Q， Feng Y Y， et al. Effects of dietary supplementation with different doses of vitamin B-complex on the growth performance and microflora composition in goats. Animal Husbandry & Veterinary Medicine， 2020， 52（7）： 47-53.
	姚志浩，梅文晴，冯宇妍，等. 复合维生素B对山羊生长性能及肠道微生物区系的影响. 畜牧与兽医， 2020， 52（7）： 47-53.
19	Caporaso J G， Lauber C L， Walters W A， et al. Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proceedings of the National Academy of Sciences of the United States of America， 2011， 108（Supple 1）： 4516-4522.
20	Youssef N， Sheik C S， Krumholz L R， et al. Comparison of species richness estimates obtained using nearly complete fragments and simulated pyrosequencing-generated fragments in 16S rRNA gene-based environmental surveys. Applied and Environmental Microbiology， 2009， 75（16）： 5227-5236.
21	Haas B J， Gevers D， Earl A M， et al. Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons. Genome Research， 2011， 21（3）： 494-504.
22	Edgar R C. UPARSE： Highly accurate OTU sequences from microbial amplicon reads. Nature Methods， 2013， 10（10）： 996.
23	Quast C， Pruesse E， Yilmaz P， et al. The SILVA ribosomal RNA gene database project： Improved data processing and web-based tools. Nucleic Acids Research， 2013， 41（D1）： 590-596.
24	O'hara E， Neves A L A， Song Y， et al. The role of the gut microbiome in cattle production and health： Driver or passenger？ Annual Review of Animal Biosciences， 2020， 8： 199-220.
25	Bäckhed F， Ding H， Wang T， et al. The gut microbiota as an environmental factor that regulates fat storage. Proceedings of the National Academy of Sciences of the United States of America， 2004， 101（44）： 15718-15723.
26	Hooper L V. Bacterial contributions to mammalian gut development. Trends in Microbiology， 2004， 12（3）： 129-134.
27	Hooper L V， Wong M H， Thelin A， et al. Molecular analysis of commensal host-microbial relationships in the intestine. Science， 2001， 291（5505）： 881-884.
28	Waters J L， Ley R E. The human gut bacteria Christensenellaceae are widespread， heritable， and associated with health. BMC Biology， 2019， 17（1）： 83.
29	Goodrich J K， Waters J L， Poole A C， et al. Human genetics shape the gut microbiome. Cell， 2014， 159（4）： 789-799.
30	Wang C， Li W， Wang H， et al. Saccharomyces boulardii alleviates ulcerative colitis carcinogenesis in mice by reducing TNF-α and IL-6 levels and functions and by rebalancing intestinal microbiota. BMC Microbiology， 2019， 19（1）： 246.
31	Egerton S， Donoso F， Fitzgerald P， et al. Investigating the potential of fish oil as a nutraceutical in an animal model of early life stress. Nutritional Neuroscience， 2020（155）： 1-23.
32	Lemaire M， Dou S， Cahu A， et al. Addition of dairy lipids and probiotic Lactobacillus fermentum in infant formula programs gut microbiota and entero-insular axis in adult minipigs. Scientific Reports， 2018， 8（1）： 1-16.
33	Özdemir F， Arslan S. Molecular characterization and toxin profiles of Bacillus spp. isolated from retail fish and ground beef. Journal of Food Science， 2019， 84（3）： 548-556.
34	Caro-Quintero A， Ritalahti K M， Cusick K D， et al. The chimeric genome of Sphaerochaeta： Nonspiral spirochetes that break with the prevalent dogma in spirochete biology. mBio， 2012， 3（3）： e00025-12.
35	Caspary W F. Physiology and pathophysiology of intestinal absorption. American Journal of Clinical Nutrition， 1992， 55（Supple 1）： 299-308.
36	Abbas B， Hayes T L， Wilson D J， et al. Internal structure of the intestinal villus： Morphological and morphometric observations at different levels of the mouse villus. Journal of Anatomy， 1989， 162： 263-273.
37	Sparks M I， Collins E N. The role of vitamin B₁ in tonus of the large intestine. American Journal of Digestive Diseases， 1935， 2（10）： 618-620.
38	Yates C A， Evans G S， Powers H J. Riboflavin deficiency： Early effects on post-weaning development of the duodenum in rats. British Journal of Nutrition， 2001， 86（5）： 593-599.
39	Berkes J， Viswanathan V K， Savkovic S D， et al. Intestinal epithelial responses to enteric pathogens： Effects on the tight junction barrier， ion transport， and inflammation. Gut， 2003， 52（3）： 439-451.
40	Zeisel M B， Dhawan P， BaumertT F. Tight junction proteins in gastrointestinal and liver disease. Gut， 2019， 68（3）： 547-561.
41	Zhang Q， Li Q， Wang C， et al. Enteropathogenic Escherichia coli changes distribution of occludin and ZO-1 in tight junction membrane microdomains in vivo. Microbial Pathogenesis， 2010， 48（1）： 28-34.
42	Wang W J， Sun D Y， Sun X F， et al. Research progress on intestinal barrier function damage and bacterial translocation. Feed Research， 2012（7）： 38-40.
	王文娟，孙冬岩，孙笑非，等. 肠道屏障功能损伤与细菌易位研究进展. 饲料研究， 2012（7）： 38-40.
43	Lu N， Wang L， Cao H， et al. Activation of the epidermal growth factor receptor in macrophages regulates cytokine production and experimental colitis. Journal of Immunology， 2014， 192（3）： 1013-1023.

项目 Item	原料 Ingredients （%）	项目 Item	营养水平 Nutrient levels^{2 ）}
精粗比The ratio of concentrate to forage	40∶60	消化能Digestible energy （MJ·kg^-1）	8.25
苜蓿青贮Alfalfa silage	60	粗蛋白Crude protein （%）	18.05
玉米 Corn	22.4	粗脂肪Ether extract （%）	3.00
麸皮Wheat bran	4.8	中性洗涤纤维Neutral detergent fiber （%）	35.07
豆粕 Soybean meal	10.8	酸性洗涤纤维Acid detergent fiber （%）	24.50
小苏打Baking soda	0.4	钙Calcium （%）	0.88
预混料 Premix^1）	1.6	总磷Total phosphorus （%）	0.35
总计 Total	100

项目 Item	原料 Ingredients （%）	项目 Item	营养水平 Nutrient levels^{2 ）}
精粗比The ratio of concentrate to forage	40∶60	消化能Digestible energy （MJ·kg^-1）	8.25
苜蓿青贮Alfalfa silage	60	粗蛋白Crude protein （%）	18.05
玉米 Corn	22.4	粗脂肪Ether extract （%）	3.00
麸皮Wheat bran	4.8	中性洗涤纤维Neutral detergent fiber （%）	35.07
豆粕 Soybean meal	10.8	酸性洗涤纤维Acid detergent fiber （%）	24.50
小苏打Baking soda	0.4	钙Calcium （%）	0.88
预混料 Premix^1）	1.6	总磷Total phosphorus （%）	0.35
总计 Total	100

基因 Gene	引物序列 Primer sequences （5′—3′）	基因序列号 GenBank accession No.
CCND	R：AAAGCCCTCTTCCATACA F：CTCCGCCTTCCCTAAC	XM_005680985.1
MKI67	R：TCAGTGAGCAGGAGGCAGTA F：GGAAATCCAGGTGACTTGCT	XM_004004769.1
Claudin 1	R：CACCCTTGGCATGAAGTGTA F：AGCCAATGAAGAGAGCCTGA	HM117762.1
Claudin 4	R：AAGGTGTACGACTCGCTGCT F：GACGTTGTTAGCCGTCCAG	HM117763.1
Occludin	R：GTTCGACCAATGCTCTCTCAG F：CAGCTCCCATTAAGGTTCCA	BC133617.1
EGFR	R：AACTGTGAGGTGGTCCTTGG F：CACTGTGTTGAGGGCAATGA	XM_005695500.1
GAPDH	R：GGGTCATCATCTCTGCACCT F：GGTCATAAGTCCCTCCACGA	HM043737.1

基因 Gene	引物序列 Primer sequences （5′—3′）	基因序列号 GenBank accession No.
CCND	R：AAAGCCCTCTTCCATACA F：CTCCGCCTTCCCTAAC	XM_005680985.1
MKI67	R：TCAGTGAGCAGGAGGCAGTA F：GGAAATCCAGGTGACTTGCT	XM_004004769.1
Claudin 1	R：CACCCTTGGCATGAAGTGTA F：AGCCAATGAAGAGAGCCTGA	HM117762.1
Claudin 4	R：AAGGTGTACGACTCGCTGCT F：GACGTTGTTAGCCGTCCAG	HM117763.1
Occludin	R：GTTCGACCAATGCTCTCTCAG F：CAGCTCCCATTAAGGTTCCA	BC133617.1
EGFR	R：AACTGTGAGGTGGTCCTTGG F：CACTGTGTTGAGGGCAATGA	XM_005695500.1
GAPDH	R：GGGTCATCATCTCTGCACCT F：GGTCATAAGTCCCTCCACGA	HM043737.1

项目 Items	指标 Index	对照组 CON	VB组 VB	P值P-value
盲肠内容物 Cecal digesta	ACE指数 ACE	1649.32±108.40	1525.63±105.40	0.45
	Chao 1指数 Chao 1	1672.20±130.08	1521.72±132.24	0.45
	香农指数 Shannon index	7.87±0.12	7.67±0.04	0.15
结肠内容物 Colonic digesta	ACE指数ACE	1383.60±106.60	1383.32±42.36	1.00
	Chao 1指数Chao 1	1343.51±104.90	1355.62±33.51	0.92
	香农指数Shannon index	7.70±0.15	7.42±0.18	0.27