短期或长期饲喂高水平豌豆纤维对猪盲肠微生物群落结构和代谢产物的影响

doi:10.11843/j.issn.0366-6964.2017.08.010

摘要/Abstract

摘要：

旨在探讨短期或长期饲喂高水平豌豆纤维（Pea fiber，PF）对猪盲肠细菌群落结构和主要代谢产物的影响。选取50头初始体重为（7.2±0.5）kg的28日龄健康断奶杜洛克×长白×大约克仔猪，按体重无差异原则随机分为2组，每组5个圈，每圈5头仔猪。处理组猪按断奶期（试验开始至断奶后30 d）、生长期（断奶后30至90 d）和育肥期（断奶后90至160 d）3个不同生理阶段猪对日粮纤维的耐受量，分别饲喂含10%、20%、30% PF的饲粮。对照组猪饲喂基础饲粮。在第一阶段（仔猪）和试验结束时（育肥猪）屠宰，采集盲肠食糜。以454高通量测序结合real-time PCR方法检测微生物群落结构。气相色谱法检测盲肠食糜中挥发性脂肪酸含量。结果表明：1）日粮中长期或短期饲喂高水平PF对猪ADFI和ADG均无显著影响（P>0.05）。2）长期饲喂PF显著增加猪盲肠中总挥发性脂肪酸含量，显著降低丙酸比例（P<0.05）；3）高通量测序分析表明，与对照组相比，短期饲喂PF的仔猪盲肠中Firmicutes门比例下降5.6%，Proteobacteria门比例增加4.3%；长期饲喂PF的肥育猪盲肠中Bacteroidetes门比例增加4.8%，Firmicutes门比例下降6.8%，采食PF后猪盲肠中存在独有优势菌属。4）Real-time PCR结果进一步证实，与对照组相比，短期饲喂PF可显著增加仔猪盲肠中总细菌、Bacteroides-Prevotella-Porphyromonas（BPP）、Enterococcus、Clostridium cluster IV的拷贝数（P<0.05），极显著增加Bacteroidetes、Lactobacillus和Desulfovibrio desulfuricans数量（P<0.01），极显著降低Firmicutes数量（P<0.01）；长期饲喂PF可极显著增加肥育猪盲肠中D.desulfuricans数量（P≤0.01），显著降低总细菌、BPP、Helicobacter-Flexispira-Wollinella（HFW）数量（P<0.05），极显著降低Enterococcus、Streptococcus和Clostridium cluster I数量（P≤0.01）。因此，猪后肠细菌群落（尤其是氢营养菌）可对日粮中高水平的PF做出迅速响应，短链脂肪酸比例的变化暗示这种菌群结构的改变与后肠微生物发酵方式的改变有关。日粮中添加高水平PF虽然可降低盲肠中条件致病菌（如Streptococcus）并提高有益菌（如Lactobacillus）的数量，但很可能不利于盲肠微生物发酵产生丁酸。

Abstract:

This study was conducted to investigate the effect of short-term or long-term feeding of high-level pea fiber (PF) on the bacterial community and metabolites in the cecum of pigs. Fifty 28 days of age healthy weaned Duroc×Landrace×Yorkshire piglets with body weight of ((7.2±0.5) kg) were selected and randomly allocated in 2 groups according to the principle of no difference in body weight in each group. There were 5 replicates (5 piglets per replicate) in each group. Pigs in the control group were given basal diets. The pigs in experimental groups were fed 10%, 20% or 30% PF diets for the post-weaning period (from experiment beginning to 30 d post-weaning) growing period (30-90 d post-weaning) and finishing period (90-160 d post-weaning), respectively. At the end of the first and the last periods, one pig from each replicate in each group was sacrificed and the cecal digesta was collected immediately. The bacterial community and the numbers of certain bacterial group were detected with 454 pyrosequencing and real-time PCR. The concentrations of volatile fatty acids in the cecal digesta were measured using gas chromatography. The results showed as follows:1) Short-term or long-term intake of high-level PF had no significant effect on the ADFI and ADG of the pigs (P>0.05). 2) The long-term feeding of PF significantly increased the concentration of total volatile fatty acids and decreased the ratio of propionate in cecal digesta of pigs(P<0.05). 3) According to the result of pyrosequencing, compared with control group, the ratio of phylum Firmicutes in cecum of piglets with short-term feeding of PF was reduced by 5.6%, while the ratio of phylum Proteobacteria was increased by 4.3%. In cecum of pigs with long-term feeding of PF, the ratio of phylum Bacteroidetes was increased by 4.8%, and the ratio of phylum Firmicutes was decreased by 6.8%. Unique bacterial species were found in cecum of pigs fed with PF supplemented diet. 4) Real-time PCR analysis confirmed that, compared with control group, short-term feed of PF significantly increased the copies of total bacteria, Bacteroides-Prevotella-Porphyromonas(BPP), Enterococcus and Clostridium cluster IV (P<0.05), and the copies of Bacteroidetes, Lactobacillus and Desulfovibrio desulfuricans (P<0.01), while the copies of Firmicutes was significantly decreased (P<0.01) in cecum of piglets. Long-term feeding of PF significantly increased the copies of D. desulfuricans (P ≤ 0.01), while decreased the copies of total bacteria, BPP, Helicobacter-Flexispira-Wollinella(HFW) (P<0.05), as well as the copies of Enterococcus, Streptococcus and Clostridium cluster I (P ≤ 0.01) in cecum of pigs. Therefore, bacteria (especially hydrogenotrophic bacteria) in hindgut of pigs can rapidly response to the high-level PF in the diet. The change of SCFAs proportion indicate that this variation of bacterial community may be involved in the altered microbial fermentation in the hindgut. Although the high level of dietary PF may reduce the abundance of some conditional pathogens such as Streptococcus and increase the abundance of some probiotics such as Lactobacillus, it may not be beneficial to the butyrate production in cecum of pigs.

中图分类号:

S828.5

罗玉衡, 陈洪, 余冰, 何军, 黄志清, 毛湘冰, 郑萍, 虞洁, 罗钧秋, 陈代文. 短期或长期饲喂高水平豌豆纤维对猪盲肠微生物群落结构和代谢产物的影响[J]. 畜牧兽医学报, 2017, 48(8): 1459-1467.

LUO Yu-heng, CHEN Hong, YU Bing, HE Jun, HUANG Zhi-qing, MAO Xiang-bing, ZHENG Ping, YU Jie, LUO Jun-qiu, CHEN Dai-wen. Short-term or Long-term Intake of High-level Pea Fiber Specifically Affects the Bacterial Community and Metabolites in the Cecum of Pigs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(8): 1459-1467.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2017.08.010

https://www.xmsyxb.com/CN/Y2017/V48/I8/1459

参考文献

[1] BÄCKHED F, MANCHESTER J K, SEMENKOVICH C F, et al. Mechanisms underlying the resistance to diet-induced obesity in germ-free mice[J]. Proc Natl Acad Sci U S A, 2007, 104(3):979-984.
[2] BROWN J M, HAZEN S L. The gut microbial endocrine organ:bacterially-derived signals driving cardiometabolic diseases[J]. Annu Rev Med, 2015, 66(1):343-359.
[3] LUO Y H, YANG C, WRIGHT A D G, et al. Responses in ileal and cecal bacteria to low and high amylose/amylopectin ratio diets in growing pigs[J]. Appl Microbiol Biotechnol, 2015, 99(24):10627-10638.
[4] HAMAKER B R, TUNCIL Y E. A perspective on the complexity of dietary fiber structures and their potential effect on the gut microbiota[J]. J Mol Biol, 2014, 426(23):3838-3850.
[5] LAMBERT J E, PARNELL J A, HAN J, et al. Evaluation of yellow pea fibre supplementation on weight loss and the gut microbiota:a randomized controlled trial[J]. BMC Gastroenterol, 2014, 14:69, doi:10.1186/1471-230X-14-69.
[6] CHE L Q, CHEN H, YU B, et al. Long-term intake of pea fiber affects colonic barrier function, bacterial and transcriptional profile in pig model[J]. Nutr Cancer, 2014, 66(3):388-399.
[7] CHEN H, MAO X B, CHE L Q, et al. Impact of fiber types on gut microbiota, gut environment and gut function in fattening pigs[J]. Anim Feed Sci Technol, 2014, 195:101-111.
[8] GUO X, XIA X, TANG R, et al. 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[J]. Lett Appl Microbiol, 2008, 47(5):367-373.
[9] IVARSSON E, ROOS S, LIU H Y, et al. Fermentable non-starch polysaccharides increases the abundance of Bacteroides-Prevotella-Porphyromonas in ileal microbial community of growing pigs[J]. Animal, 2014, 8(11):1777-1787.
[10] RINTTILÄ T, KASSINEN A, MALINEN E, et al. Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR[J]. J Appl Microbiol, 2004, 97(6):1166-1177.
[11] VAN DYKE M I, MCCARTHY A J. Molecular biological detection and characterization of Clostridium populations in municipal landfill sites[J]. Appl Environ Microbiol, 2002, 68(4):2049-2053.
[12] CASTILLO M, MARTÍN-ORU'E S M, MANZANILLA E G, et al. Quantification of total bacteria, enterobacteria and lactobacilli populations in pig digesta by real-time PCR[J]. Vet Microbiol, 2006, 114(1-2):165-170.
[13] RYU H, GRIFFITH J F, KHAN I U H, et al. Comparison of gull feces-specific assays targeting the 16S rRNA genes of Catellicoccus marimammalium and Streptococcus spp.[J]. Appl Environ Microbiol, 2012, 78(6):1909-1916.
[14] BARTOSCH S, FITE A, MACFARLANE G T, et al. Characterization of bacterial communities in feces from healthy elderly volunteers and hospitalized elderly patients by using real-time PCR and effects of antibiotic treatment on the fecal microbiota[J]. Appl Environ Microbiol, 2004, 70(6):3575-3581.
[15] TURNBAUGH P J, LEY R E, MAHOWALD M A, et al. An obesity-associated gut microbiome with increased capacity for energy harvest[J]. Nature, 2006, 444(7122):1027-1031.
[16] DE FILIPPO C, CAVALIERI D, DI PAOLA M, et al. Impact of diet in shaping gut microbiota revealed by a comparative study in children from Europe and rural Africa[J]. Proc Natl Acad Sci U S A, 2010, 107(33):14691-14696.
[17] LA-ONGKHAM O, NAKPHAICHIT M, LEELAVATCHARAMAS V, et al. Distinct gut microbiota of healthy children from two different geographic regions of Thailand[J]. Arch Microbiol, 2015, 197(4):561-573.
[18] DEN BESTEN G, VAN EUNEN K, GROEN A K, et al. The role of short-chain fatty acids in the interplay between diet, gut microbiota, and host energy metabolism[J]. J Lipid Res, 2013, 54(9):2325-2340.
[19] JENSEN B B, JØRGENSEN H. Effect of dietary fiber on microbial activity and microbial gas production in various regions of the gastrointestinal tract of pigs[J]. Appl Environ Microbiol, 1994, 60(6):1897-1904.
[20] BOETS E, DEROOVER L, LUYPAERTS A, et al. Utilization of colonic derived propionate in gluconeogenesis:an in vivo stable isotope study in humans[J]. Acta Gastroenterol Belg, 2015, 78(1).
[21] REY F E, GONZALEZ M D, CHENG J, et al. Metabolic niche of a prominent sulfate-reducing human gut bacterium[J]. Proc Natl Acad Sci U S A, 2013, 110(33):13582-13587.
[22] TROMPETTE A, GOLLWITZER E S, YADAVA K, et al. Gut microbiota metabolism of dietary fiber influences allergic airway disease and hematopoiesis[J]. Nat Med, 2014, 20(2):159-166.
[23] BULTMAN S J. Molecular pathways:gene-environment interactions regulating dietary fiber Induction of proliferation and apoptosis via butyrate for cancer prevention[J]. Clin Cancer Res, 2014, 20(4):799-803.
[24] PUERTOLLANO E, KOLIDA S, YAQOOB P. Biological significance of short-chain fatty acid metabolism by the intestinal microbiome[J]. Curr Opin Clin Nutr, 2014, 17(2):139-144.
[25] PATTEN G S, KERR C A, DUNNE R A, et al. Resistant starch alters colonic contractility and expression of related genes in rats fed a western diet[J]. Digest Dis Sci, 2015, 60(6):1624-1632.