五指山猪和长白猪断奶仔猪肠道免疫性能及微生物多样性比较分析

doi:10.16431/j.cnki.1671-7236.2023.08.009

摘要/Abstract

摘要： 【目的】探讨早期断奶对五指山猪和长白猪小肠肠道免疫性能及微生物多样性的影响差异。【方法】选择生长发育及营养状况良好、28日龄断奶的长白猪与五指山猪各6头,于50日龄采集小肠各段组织、内容物、黏膜,依次采用石蜡切片(HE染色)、16S rDNA V3-V4区基因高通量测序、实时荧光定量PCR测定,比较两品种间肠道形态学、肠道菌群及肠道黏膜免疫相关基因mRNA相对表达量的差异。【结果】①五指山猪空肠绒毛高度显著低于长白猪(P＜0.05)。②长白猪十二指肠与回肠微生物多样性均较五指山猪更丰富(P＜0.05)。③在门水平上,五指山猪空肠、回肠中厚壁菌门(Firmicutes)的丰度均显著高于长白猪,但变形菌门(Proteobacteria)出现相反的表达趋势(P＜0.05)。在属水平上,五指山猪回肠中乳杆菌属(Lactobacillus)的丰度显著高于长白猪(P＜0.05)。④黏膜免疫相关基因表达量的检测结果显示,长白猪十二指肠中抑炎因子白细胞介素10(IL-10)和紧密连接蛋白闭锁小带蛋白1(ZO1)的mRNA表达量均显著高于五指山猪(P＜0.05);而空肠与回肠中促炎因子肿瘤坏死因子-α(TNF-α)的mRNA表达量均显著低于五指山猪(P＜0.05)。【结论】与长白猪相比,早期断奶后五指山猪小肠肠道免疫性能及微生物多样性较差,可适当延长五指山猪的断奶日龄来减少仔猪腹泻率,以保证断奶仔猪的成活率。

关键词: 五指山猪; 长白猪; 早期断奶; 免疫性能; 肠道微生物多样性

Abstract: 【Objective】 To explore the difference of effects of early weaning on intestinal immunity and microbial diversity in Wuzhishan and Landrace pigs.【Method】 Six 50-day-old Landrace pigs and six Wuzhishan pigs (weaned at 28-day-old) with good growth,development and nutritional status were selected to collect the tissues,contents and mucosa of each segment of the small intestine.Paraffin section (HE staining),16S rDNA V3-V4 gene high-throughput sequencing and Real-time PCR were used to compare the differences between the two breeds in intestinal morphology,intestinal flora and the relative mRNA expression of intestinal mucosal immune-related genes.【Result】 ① The jejunal villus height of Wuzhishan pigs was significantly lower than that of Landrace pigs (P＜0.05).②The intestinal microbial diversity of Landrace pigs was more abundant in duodenum and ileum than that of Wuzhishan pigs (P＜0.05).③At the phylum level,the abundance of Firmicutes in the jejunum and ileum of Wuzhishan pigs was significantly higher than that of Landrace pigs,but Proteobacteria showed a reverse expression trend (P＜0.05).At the genus level,the abundance of Lactobacillus in ileum of Wuzhishan pigs was significantly higher than that of Landrace pigs (P＜0.05).④The detection results of mucosal immune related gene expression showed that the mRNA expression levels of anti-inflammatory factor IL-10 and tight junction protein ZO1 in duodenum of Landrace pigs were significantly higher than those of Wuzhishan pigs (P＜0.05),while the mRNA expression levels of pro-inflammatory factor TNF-α in jejunum and ileum were significantly lower than those in Wuzhishan pigs (P＜0.05).【Conclusion】 Compared with Landrace pigs,Wuzhishan pigs had poor intestinal immune performance and microbial diversity after early weaning.The weaning age of Wuzhishan pigs could be appropriately extended to reduce the diarrhea rate and ensure the survival rate of weaned piglets.

Key words: Wuzhishan pig; Landrace pig; early weaning; immunity; gut microbial diversity

中图分类号:

S828

张婷, 魏立民, 刘光亮, 王峰, 晁哲, 任钰为, 刘海隆, 郑心力, 黄丽丽, 孙瑞萍. 五指山猪和长白猪断奶仔猪肠道免疫性能及微生物多样性比较分析[J]. 中国畜牧兽医, 2023, 50(8): 3113-3122.

ZHANG Ting, WEI Limin, LIU Guangliang, WANG Feng, CHAO Zhe, REN Yuwei, LIU Hailong, ZHENG Xinli, HUANG Lili, SUN Ruiping. Comparative Analysis of Intestinal Microbial Diversity and Immunity in the Small Intestine of Weaned Piglets from Wuzhishan Pigs and Landrace Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3113-3122.

参考文献

[1] MONTAGNE L,BOUDRY G,FAVIER C,et al.Main intestinal markers associated with the changes in gut architecture and function in piglets after weaning[J].The British Journal of Nutrition,2007,97(1):45-57.
[2] YIN J,WU M M,XIAO H,et al.Development of an antioxidant system after early weaning in piglets[J].Journal of Animal Science,2014,92(2):612-619.
[3] 余冰,张克英,郑萍,等.猪营养与肠道健康[J].中国畜牧杂志,2010,46(15):73-76. YU B,ZHANG K Y,ZHENG P,et al.Pig nutrition and intestinal health[J].Chinese Journal of Animal Science,2010,46(15):73-76.(in Chinese)
[4] 郑红梅.蛹虫草对育肥猪肠道形态和屏障功能的影响[D].杨凌:西北农林科技大学,2020. ZHENG H M.Effects of Cordyceps militaris on intestinal morphology and barrier function in finishing pigs[D].Yangling:Northwest A&F University,2020.(in Chinese)
[5] CAMPBELL J M,CRENSHAW J D,POLO J J,et al.The biological stress of early weaned piglets[J].Journal of Animal Science and Biotechnology,2013,4(2):124-127.
[6] HAMPSON D J.Alterations in piglet small intestinal structure at weaning[J].Research in Veterinary Science,1986,40(1):32-40.
[7] CASTILLO M,MARTÍN-OR AU'G1 E S M,NOFRARÍAS M,et al.Changes in caecal microbiota and mucosal morphology of weaned pigs[J].Veterinary Microbiology,2007,124(3-4):239-247.
[8] 曹婷.五指山猪和长白猪肌肉发育差异的初步研究[D].海口:海南大学,2013. CAO T.A preliminary study on the difference of muscle development between Wuzhishan pig and Landrace pig[D].Haikou:Hainan University,2013.(in Chinese)
[9] 李吉涛.五指山猪近交系矮小性分子遗传基础研究[D].北京:中国农业科学院,2007. LI J T.Molecular genetic basis of dwarfism in Wuzhishan pig inbred lines[D].Beijing:Chinese Academy of Agricultural Sciences,2007.(in Chinese)
[10] 单立莉,张敏,苗志国,等.Myogenin mRNA丰度在金华猪和长白猪背最长肌中的表达[J].中国兽医学报,2009,29(3):374-388. SHAN L L,ZHANG M,MIAO Z G,et al.Expression of myogenin mRNA abundance in longest dorsal muscle of Jinhua pig and Landrace pig[J].Chinese Journal of Veterinary Medicine,2009,29(3):374-388.(in Chinese)
[11] 陈逸,董丽,喻礼怀.肠道微生物区系对断奶仔猪肠道黏膜免疫功能的调节作用研究进展[J].中国饲料,2019,13:9-12. CHEN Y,DONG L,YU L H.Research progress on the regulatory effect of intestinal microflora on intestinal mucosal immune function in weaned piglets[J].China Feed,2019,13:9-12.(in Chinese)
[12] 冯跃进.热应激对猪和大鼠肠道结构和功能的影响及其修复机制[D].北京:中国农业科学院,2014. FENG Y J.Effects of heat stress on intestinal structure and function of pigs and rats and its repair mechanism[D].Beijing:Chinese Academy of Agricultural Sciences,2014.(in Chinese)
[13] 高鹏飞,梁雅旭,宰小玉,等.山西黑猪与大白猪肠道结构发育的比较学研究[J].国外畜牧学-猪与禽,2018,38(5):26-29. GAO P F,LIANG Y X,ZAI X Y,et al.A comparative study on the intestinal structure and development of Shanxi Black and Large White pigs[J].Animal Science Abroad-Pigs and Poultry,2018,38(5):26-29.(in Chinese)
[14] 贺晓玉.蛹虫草菌固体发酵五味子药渣工艺的优化及其产物对断奶仔猪肠道形态结构和功能的影响研究[D].成都:四川农业大学,2014. HE X Y.Optimization of solid fermentation of Schisandra chinensis dregs by Cordyceps militaris and the effect of its products on the intestinal morphology and function of weaned piglets[D].Chengdu:Sichuan Agricultural University,2014.(in Chinese)
[15] 赵铭伟,奉玲丽,张琰芳,等.鞣花酸对断奶仔猪小肠黏膜形态及肠黏膜屏障功能的影响[J].黑龙江畜牧兽医,2020,11:126-130. ZHAO M W,FENG L L,ZHANG Y F,et al.Effect of ellagic acid on intestinal mucosal morphology and intestinal mucosal barrier function in weaned piglets[J].Heilongjiang Animal Science and Veterinary Medicine,2020,11:126-130.(in Chinese)
[16] CHEN X,SONG P,FAN P,et al.Moderate dietary protein restriction optimized gut microbiota and mucosal barrier in growing pig model[J].Frontiers in Cellular and Infection Microbiology, 2018,18(8):246-246.
[17] 龚月生,张维,曹雨莉,等.日粮蛋白质水平对早期断奶仔猪生产性能、粪便微生物及血液生化指标的影响[A].全国猪营养学术研讨会[C].2007. GONG Y S,ZHANG W,CAO Y L,et al.Effect of dietary protein level on production performance,fecal microorganisms and blood biochemical indexes of early weaned piglets[A].National Symposium on Pig Nutrition[C].2007.(in Chinese)
[18] 李卓君,陈春香,钟小菊,等.猪肠道微生物组成影响因素及其对重要经济性状的影响研究进展[J].中国畜牧兽医,2022,49(7):2557-2566. LI Z J,CHEN C X,ZHONG X J,et al.Research progress on influencing factors of pig intestinal microbial composition and their effects on important economic traits[J].China Animal Husbandry&Veterinary Medicine,2022,49(7):2557-2566.(in Chinese)
[19] DIAO H,YAN H L,XIAO Y,et al.Intestinal microbiota could transfer host gut characteristics from pigs to mice[J].BMC Microbiology,2016,16(1):238-253.
[20] BERGAMASCHI M,TIEZZI F,HOWARD J,et al.Gut microbiome composition differences among breeds impact feed efficiency in swine[J].Microbiome,2020,8(1):110-127.
[21] QI R,SUN J,QIU X,et al.The intestinal microbiota contributes to the growth and physiological state of muscle tissue in piglets[J].Scientific Reports,2021,11(1):11237.
[22] FRANK D N,AMAND A,FELDMAN R A,et al.Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases[J].Proceedings of the National Academy of Sciences of the United States of America, 2007,104(34):13780-13785.
[23] 范沛昕.低蛋白日粮对断奶仔猪和育肥猪肠道微生物区系的影响[D].北京:中国农业大学,2016. FAN P X.Effect of low protein diet on intestinal microflora of weaned piglets and finishing pigs[D].Beijing:China Agricultural University,2016.(in Chinese)
[24] ZHANG L,WU W,LEE Y K,et al.Spatial heterogeneity and co-occurrence of mucosal and luminal microbiome across swine intestinal tract[J].Frontiers in Microbiology,2018,1(26):48-61.
[25] IVARSSON E,ROOS,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.
[26] REGMI P R,METZLER-ZEBELI B U,GANZLE M G,et al.Starch with high amylose content and low in vitro digestibility increases intestinal nutrient flow and microbial fermentation and selectively promotes bifidobacteria in pigs[J].The Journal of Nutrition,2011,141(7):1273-1280.
[27] GUO X,XIA X,TANG R,et al.Real-time PCR quantification of the predominant bacterial divisions in the distal gut of Meishan and Landrace pigs[J].Anaerobe,2008,14(4):224-228.
[28] 刘亚丹,左周,秦军军,等.仔猪不同生长阶段粪便微生物多样性分析[J].国外畜牧学-猪与禽,2020,40(6):22-29. LIU Y D,ZUO Z,QIN J J,et al.Analysis of fecal microbial diversity of piglets at different growth stages[J].Animal Science Abroad-Pigs and Poultry,2020,40(6):22-29.(in Chinese)
[29] XIN J G,ZENG D,WANG H S,Probiotic Lactobacillus johnsonii BS15 promotes growth performance,intestinal immunity,and gut microbiota in piglets[J].Probiotics and Antimicrobial Proteins,2019,12(1):184-193.
[30] BEAUMONT M,GOODRICH J K,JACKSON M A,et al.Heritable components of the human fecal microbiome are associated with visceral fat[J].Genome Biology,2016,17(1):61-67.
[31] LIM M Y,SONG E J,KANG K S,et al.Age-related compositional and functional changes in micro-pig gut microbiome[J].GeroScience,2019,41(6):935-944.
[32] DONG L,ZHONG X,HE J,et al.Supplementation of tributyrin improves the growth and intestinal digestive and barrier functions in intrauterine growth-restricted piglets[J].Clinical Nutrition, 2016,35(2):399-407.
[33] 冯宝宝,赵莹,马晓宇,等.不同教槽料对断奶仔猪肠道黏膜免疫相关基因表达的影响[J].河南农业科学,2020,9(18):143-152. FENG B B,ZHAO Y,MA X Y,et al.Effect of different teaching materials on intestinal mucosal immune related gene expression of weanling piglets[J].Journal of Henan Agricultural Sciences,2020,9(18):143-152.(in Chinese)
[34] WONDRA K J,HANCOCK J D,BEHNKE K C,et al.Effects of particle size of barley on intestinal morphology,growth performance and nutrient digestibility in pigs[J].Journal of Animal Science,2007,20(11):1738-1745.
[35] PIÉ S,LALLōS J,BLAZY F,et al.Weaning is associated with an upregulation of expression of inflammatory cytokines in the intestine of piglets[J].The Journal of Nutrition,2004,134(3):641-647.
[36] HU C H,XIAO K,LUAN Z S,et al.Early weaning increases intestinal permeability,alters expression of cytokine and tight junction proteins,and activates mitogen-activated protein kinases in pigs[J].Journal of Animal Science, 2013,91(3):1094-1101.
[37] 郑兰,李亚雄.抗生素及其替代物在畜禽肠道内的作用及其对肠道微生物和形态结构的影响[J].国外畜牧学-猪与禽,2017,37(2):57-60.ZHENG L,LI Y X.The role of antibiotics and their substitutes in the intestinal tract of livestock and poultry and their effects on intestinal microorganisms and morphology[J].Animal Science Abroad-Pig and Poultry,2017,37(2):57-60.(in Chinese)