猪营养与饲料 SWINE NUTRITION AND FEED

纯种柯乐猪与巴×柯杂交猪肠道菌群结构的研究

  • 杨莲 ,
  • 燕志宏 ,
  • 黄维江 ,
  • 杨仕钰 ,
  • 吴光松 ,
  • 顾丽菊 ,
  • 林鹏飞 ,
  • 杨蓉 ,
  • 李平 ,
  • 任丽群 ,
  • 张芸
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  • 1. 贵州大学高原山地动物遗传育种与繁殖教育部重点实验室, 贵阳 550000;
    2. 贵州大学贵州省动物遗传育种与繁殖重点实验室, 贵阳 550000;
    3. 贵州大学动物科学学院, 贵阳 550000;
    4. 开阳县畜禽品种改良站, 贵阳 550000;
    5. 贵州优农谷生态产业有限公司, 毕节 551700;
    6. 贵州省物资现代物流集团有限责任公司, 贵阳 550000;
    7. 贵州省畜禽种质测定中心, 贵阳 550000;
    8. 贵州省农业区域经济发展中心, 贵阳 550000
杨莲(1995-),女,贵州安顺人,硕士研究生,从事动物营养学与肠道菌群研究。E-mail:2076819301@qq.com

收稿日期: 2020-09-07

  网络出版日期: 2021-03-18

基金资助

贵州省科技厅农业攻关项目(黔农育专字[2016]009号)

Study on Intestinal Microflora Structure of Pure-Bred Kele Pigs and Berkshire×Kele Hybrid Pigs

  • YANG Lian ,
  • YAN Zhihong ,
  • HUANG Weijiang ,
  • YANG Shiyu ,
  • WU Guangsong ,
  • GU Liju ,
  • LIN Pengfei ,
  • YANG Rong ,
  • LI Ping ,
  • REN Liqun ,
  • ZHANG Yun
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  • 1. Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountain Region, Ministry of Education, Guizhou University, Guiyang 550000, China;
    2. Guizhou Province Key Laboratory of Animal Genetics, Breeding and Reproduction, Guizhou University, Guiyang 550000, China;
    3. College of Animal Science, Guizhou University, Guiyang 550000, China;
    4. Kaiyang County Livestock and Poultry Variety Improvement Station, Guiyang 550000, China;
    5. Guizhou Younong Valley Ecological Industry Co., Ltd., Bijie 551700, China;
    6. Guizhou Material Modern Logistics Group Co., Ltd., Guiyang 550000, China;
    7. Guizhou Provincial Livestock and Poultry Germplasm Testing Center, Guiyang 550000, China;
    8. Guizhou Agricultural Regional Economic Development Center, Guiyang 550000, China

Received date: 2020-09-07

  Online published: 2021-03-18

Supported by

 

摘要

本试验对纯种柯乐猪与巴×柯杂交猪肠道菌群结构特性及差异性进行了研究,旨在从猪肠道微生物的角度揭示纯种柯乐猪耐粗饲特性及巴×柯杂交猪对青绿饲料的适应能力,为后期微生物添加剂的设计、柯乐猪杂交利用及标准化养殖提供依据。纯种柯乐猪和巴×柯杂交猪在同等条件下养殖,达屠宰体重后(100 kg左右),随机各选3头屠宰,取十二指肠、空肠、回肠、结肠内容物,进行高通量测序和生物学信息分析。结果显示:纯种柯乐猪与巴×柯杂交猪各肠段Alpha多样性指数差异不显著(P>0.05);Beta多样性分析显示,2个品种猪大部分样品交叉聚集到一起;肠道菌群结构分析显示,纯种柯乐猪和巴×柯杂交猪小肠阶段以厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)和放线菌门(Actinobacteria)为优势菌门,到了结肠阶段则是以拟杆菌门(Bacteroidetes)和厚壁菌门为优势菌门;纯种柯乐猪与巴×柯杂交猪结肠中有大量的纤维分解菌属,巴×柯杂交猪结肠中月形单胞菌属(Selenomonas)和脱硫弧菌属(Desulfovibrio)的相对丰度显著或极显著高于纯种柯乐猪(P<0.05或P<0.01),而消化球菌属(Peptococcus)、聚乙酸菌属(Acetitomaculum)、Leeia的相对丰度则显著低于纯种柯乐猪(P<0.05);LEfSe分析后发现纯种柯乐猪结肠中富集着大量的产短链脂肪酸菌,其中纤维降解菌有聚乙酸菌属、瘤胃球菌科的不可培养瘤胃细菌4C0d_17(uncultured_rumen_bacterium_4C0d_17)、Leeia、拟杆菌纲、拟杆菌目,与纤维降解菌起协同作用的菌群有考拉杆菌属(Phascolarctobacterium)、考拉杆菌属的uncultured_Veillonellaceae_bacterium;巴×柯杂交猪结肠中富集的纤维降解菌有月形单胞菌属、柔嫩梭菌(Clostridium leptum)。上述结果表明,在整个肠道菌群结构上,纯种柯乐猪和巴×柯杂交猪的优势菌在类别上非常接近,体现了环境相同效应,纯种柯乐猪和巴×柯杂交猪的肠道菌群结构相对稳定且相似度高;在属水平上,纯种柯乐猪和巴×柯杂交猪的结肠中纤维降解菌表现出差异,纯种柯乐猪结肠内纤维降解菌的比例较高,说明引入外源血对肠道菌群结构有一定的影响,巴×柯杂交猪对粗纤维的消化能力较弱于柯乐猪。

本文引用格式

杨莲 , 燕志宏 , 黄维江 , 杨仕钰 , 吴光松 , 顾丽菊 , 林鹏飞 , 杨蓉 , 李平 , 任丽群 , 张芸 . 纯种柯乐猪与巴×柯杂交猪肠道菌群结构的研究[J]. 动物营养学报, 2021 , 33(3) : 1359 -1371 . DOI: 10.3969/j.issn.1006-267x.2021.03.019

Abstract

This experiment studied the structural characteristics and differences of intestinal microflora of pure-bred Kele pigs and Berkshire×Kele hybrid pigs, aiming to reveal the rough feed tolerance characteristics of Kele pigs and the ability to adapt to green feed of Berkshire×Kele hybrid pigs from the perspective of pig gut microbes, and to provide a basis for the design of microbial additives in the later stage, the cross-utilization of Kele pigs and standardized breeding. Pure-bred Kele pigs and Berkshire×Kele hybrid pigs were raised under the same conditions. After reaching the slaughter weight (about 100 kg), three pigs were randomly selected for slaughter, and the contents of the duodenum, jejunum, ileum and colon were taken to perform high-throughput sequencing and biological information analysis. The results showed that there were no significant differences in Alpha diversity indexes in each intestinal segment between Berkshire×Kele hybrid pigs and pure-bred Kele pigs (P>0.05). In Beta diversity analysis, most samples in the two breeds of pigs crossed and gathered together. Intestinal microflora structure analysis showed that, Firmicutes, Proteobacteria, Actinobacteria were the dominant bacterial phyla in the stage of small intestine of Berkshire×Kele hybrid pigs and pure-bred Kele pigs, and Bacteroidetes and Firmicutes were the dominant bacterial phyla in the stage of colon. There were a lot of cellulose decomposition microbes in the colon of Berkshire×Kele hybrid pigs and pure-bred Kele pigs. The relative abundances of Selenomonas and Desulfovibrio in the colon of Berkshire×Kele hybrid pigs were significantly higher than those of pure-bred Kele pigs (P<0.05 or P<0.01), and the relative abundances of Peptococcus, Acetitomaculum and Leeia were significantly lower than those of pure-bred Kele pigs (P<0.05). LEfSe analysis found that a lot of short chain fatty acid producing bacteria enriched in the pure-bred Kele pigs’ colon. Among them, Acetitomaculum, uncultured_rumen_bacterium_4C0d_17, Leeia, Bacteroidia and Bacteroidales belonged to fiber-degrading bacteria, and some bacteria had synergistic effects with the fiber-degrading bacteria, they were Phascolarctobacterium and uncultured_Veillonellaceae_bacterium. The fiber-degrading bacteria such as Selenomonas and Clostridium leptum enriched in the Berkshire×Kele hybrid pigs’ colon. Above results indicate that, in the structure of entire intestinal microflora, the type of dominant bacteria in pure-bred Kele pigs and Berkshire×Kele hybrid pigs is almost similar, which reflects the same environmental effects, and the structure of the intestinal microflora of pure-bred Kele pigs and Berkshire×Kele hybrid pigs is relatively stable and the similarity is high; at genus level, the fiber-degrading bacteria show some differences between pure-bred Kele pigs and Berkshire×Kele hybrid pigs, and fiber-degrading bacteria accounts for a higher proportion in cecal microflora of pure-bred Kele pigs, which indicates that the introduction of exogenous blood has a certain impact on the intestinal microflora, and Berkshire×Kele hybrid pigs are weaker in digesting crude fiber than pure-bred Kele pigs.

参考文献

[1] 陈润生.猪生产学[M].北京:中国农业出版社,1995. CHEN R S.Pig production[M].Beijing:China Agriculture Press,1995.(in Chinese)
[2] 张伟力,顾平生.柯乐猪肉切块品质点评[J].养猪,2013(5):63-64. ZHANG W L,GU P S.Kele pork cut quality review[J].Swine Production,2013(5):63-64.(in Chinese)
[3] 杨蓉,林鹏飞,吴光松,等.不同屠宰期柯乐猪肉品质的差异研究[J].黑龙江畜牧兽医,2019(14):44-46. YANG R,LIN P F,WU G S,et al.Study on the difference of Kele pork quality in different slaughter periods[J].Heilongjiang Animal Science and Veterinary Medicine,2019(14):44-46.(in Chinese)
[4] 杨蓉,林鹏飞,吴光松,等.饲养方式对不同体重柯乐猪屠宰性能及胴体品质的影响[J].黑龙江畜牧兽医,2019(13):65-69. YANG R,LIN P F,WU G S,et al.Effects of feeding methods on the slaughter performance and carcass quality of Kele pigs of different weights[J].Heilongjiang Animal Science and Veterinary Medicine,2019(13):65-69.(in Chinese)
[5] 张伟力,张似青,张磊彪.巴克夏在中国的历史辉煌与现代美系巴克夏猪性能初报[J].养猪,2010(3):68-72. ZHANG W L,ZHANG S Q,ZHANG L B.Preliminary report on the historical brilliance of Bakxia in China and the performance of modern American Berkshire pigs[J].Swine Production,2010(3):68-72.(in Chinese)
[6] 李玉莲,吴买生,王建伟,等.大约克、杜洛克、巴克夏猪肥育和胴体性状比较分析[J].家畜生态学报,2018,39(3):44-47. LI Y L,WU M S,WANG J W,et al.Analysis on fattening performance and carcass traits of Yorkshire,Duroc and Berkshire pigs[J].Journal of Domestic Animal Ecology,2018,39(3):44-47.(in Chinese)
[7] YANG L N,BIAN G R,SU Y,et al.Comparison of faecal microbial community of Lantang,Bama,Erhualian,Meishan,Xiaomeishan,Duroc,Landrace,and Yorkshire sows[J].Asian-Australasian Journal of Animal Sciences,2014,27(6):898-906.  
[8] SUN L J,MA L J,MA Y B,et al.Insights into the role of gut microbiota in obesity:pathogenesis,mechanisms,and therapeutic perspectives[J].Protein & Cell,2018,9(5):397-403.  
[9] 杨柳,张邑帆,郑华,等.荣昌、长白、杜洛克猪肠道微生物ERIC-PCR-DGGE指纹图谱比较分析[J].家畜生态学报,2011,32(5):21-25. YANG L,ZHANG Y F,ZHENG H,et al.Intestinal microbial community and its fingerprint comparative analysis between Rongchang,Landrace and Duroc using ERIC-PCR-DGGE[J].Journal of Domestic Animal Ecology,2011,32(5):21-25.(in Chinese)
[10] PIEPER R,VAHJEN W,ZENTEK J.Dietary fibre and crude protein:impact on gastrointestinal microbial fermentation characteristics and host response[J].Animal Production Science,2015,55(12):1367-1375.  
[11] TIAN L M,BRUGGEMAN G,VAN DEN BERG M,et al.Effects of pectin on fermentation characteristics,carbohydrate utilization,and microbial community composition in the gastrointestinal tract of weaning pigs[J].Molecular Nutrition & food Research,2017,61(1):1600186.
[12] 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].Animal Feed Science and Technology,2014,195:101-111.
[13] YANG Y X,MU C L,LUO Z,et al.Bromochloromethane,a methane analogue,affects the microbiota and metabolic profiles of the rat gastrointestinal tract[J].Applied and Environmental Microbiology,2016,82(3):778-787.  
[14] LOOFT T,JOHNSON T A,ALLEN H K,et al.In-feed antibiotic effects on the swine intestinal microbiome[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(5):1691-1696.  
[15] ZHOU X L,KONG X F,LIAN G Q,et al.Dietary supplementation with soybean oligosaccharides increases short-chain fatty acids but decreases protein-derived catabolites in the intestinal luminal content of weaned Huanjiang mini-piglets[J].Nutrition Research,2014,34(9):780-788.  
[16] LEE S H,INGALE S L,KIM J S,et al.Effects of dietary supplementation with Bacillus subtilis LS 1-2 fermentation biomass on growth performance,nutrient digestibility,cecal microbiota and intestinal morphology of weanling pig[J].Animal Feed Science and Technology,2014,188:102-110.
[17] 胡远亮.利用分子生物技术研究益生菌对断奶仔猪生长及粪便菌群的影响[D].博士学位论文.武汉:华中农业大学,2014:25-87. HU Y L.Investigation of probiotics on the growth and fecal bacterial community of weaned piglets with molecular biotechnology[D].Ph.D.Thesis.Wuhan:Huazhong Agricultural University,2014:25-87.(in Chinese)
[18] YI J Q,PIAO X S,LI Z C,et al.The effects of enzyme complex on performance,intestinal health and nutrient digestibility of weaned pigs[J].Asian-Australasian Journal of Animal Sciences,2013,26(8):1181-1188.  
[19] 吴超,张莉,吴跃明,等.中草药添加剂对早期断奶仔猪生长性能和肠道菌群的影响[J].中国畜牧杂志,2010,46(3):31-35,52. WU C,ZHANG L,WU Y M,et al.Effects of Chinese herb additives on growth performance and intestinal microflora of weaned piglets[J].Chinese Journal of Animal Science,2010,46(3):31-35,52.(in Chinese)
[20] 王宪龙,王元虎,李少青,等.野莱F1代猪胴体品质、肉质性状和肌内脂肪酸组成的研究[J].山东农业大学学报(自然科学版),2012,43(1):62-67. WANG X L,WANG Y H,LI S Q,et al.The studies on carcass quality,meat character and intramuscular fatty acids composition of F1 hybrid with boar and Laiwu pigs[J].Journal of Shandong Agricultural University (Natural Science Edition),2012,43(1):62-67.(in Chinese)
[21] ROTHSCHILD D,WEISSBROD O,BARKAN E,et al.Environment dominates over host genetics in shaping human gut microbiota[J].Nature,2018,555(7695):210-215.  
[22] 徐娥,杨华,刘秀婷,等.大约克猪肠道不同部位的菌群结构和短链脂肪酸含量研究[J].动物营养学报,2019,31(10):4509-4518. XU E,YANG H,LIU X T,et al.Study on bacterial community structure and short chain fatty acid content in different parts of intestines of Yorkshire pigs[J].Chinese Journal of Animal Nutrition,2019,31(10):4509-4518.(in Chinese)
[23] 王芳芳,刁华杰,王志龙,等.中草药添加剂对断奶仔猪肠道菌群与生长性能的影响[J].中国畜牧杂志,2017,53(3):121-124. WANG F,DIAO J H,WANG Z L,et al.Effects of herbal additives on performance and intestinal microflora of weaned piglets[J].Chinese Journal of Animal Science,2017,53(3):121-124.(in Chinese)
[24] 郭晓红,郭玉龙,刘亚丹,等.仔猪不同发育阶段结肠微生物菌群特征分析[J].畜牧兽医学报,2019,50(9):1759-1774. GUO X H,GUO Y L,LIU Y D,et al.Characteristics of intestinal microflora of colon at different developmental stages in piglets[J].Journal of Domestic Animal Ecology,2019,50(9):1759-1774.(in Chinese)
[25] GAO P F,LIU Y D,YE B Y,et al.A comparison of dynamic distributions of intestinal microbiota between Large White and Chinese Shanxi Black pigs[J].Archives of Microbiology,2019,201(3):357-367.  
[26] KONSTANTINOV S R,FAVIER C F,ZHU W Y,et al.Microbial diversity studies of the porcine gastrointestinal ecosystem during weaning transition[J].Animal Research,2004,53(4):317-324.  
[27] DE AGÜERO M G,GANAL-VONARBURG S C,FUHRER T,et al.The maternal microbiota drives early postnatal innate immune development[J].Science,2016,351(6279):1296-1302.  
[28] MUSSO G,GAMBINO R,CASSADER M.Interactions between gut microbiota and host metabolism predisposing to obesity and diabetes[J].Annual Review of Medicine,2011,62:361-380.
[29] KONG X F,ZHOU X L,LIAN G Q,et al.Dietary supplementation with chitooligosaccharides alters gut microbiota and modifies intestinal luminal metabolites in weaned Huanjiang mini-piglets[J].Livestock Science,2014,160:97-101.
[30] 杨伟平.藏猪肠道细菌群落组成与纤维素分解菌的研究[D].博士学位论文.杨凌:西北农林科技大学,2015. YANG W P.A study on the bacteria community and the cellulolytic bacterium in Tibetan pigs[D].Ph.D.Thesis.Yangling:Northwest A& F University,2015.(in Chinese)
[31] LAU S K P,TENG J L L,HO C T,et al.Differential microbial communities of omnivorous and herbivorous cattle in southern China[J].Computational and Structural Biotechnology Journal,2018,16:54-60.
[32] BEARDS E,TUOHY K,GIBSON G.Bacterial,SCFA and gas profiles of a range of food ingredients following in vitro fermentation by human colonic microbiota[J].Anaerobe,2010,16(4):420-425.  
[33] MACKIE R I,AMINOV R I,HU W P,et al.Ecology of uncultivated Oscillospira species in the rumen of cattle,sheep,and reindeer as assessed by microscopy and molecular approaches[J].Applied and Environmental Microbiology,2003,69(11):6808-6815.  
[34] STEVENSON D M,WEIMER P J.Dominance of Prevotella and low abundance of classical ruminal bacterial species in the bovine rumen revealed by relative quantification real-time PCR[J].Applied Microbiology and Biotechnology,2007,75(1):165-174.  
[35] YANG H,XIAO Y P,WANG J J,et al.Core gut microbiota in Jinhua pigs and its correlation with strain,farm and weaning age[J].Journal of Microbiology,2018,56(5):346-355.  
[36] LOOFT T,ALLEN H K,CANTAREL B L,et al.Bacteria,phages and pigs:the effects of in-feed antibiotics on the microbiome at different gut locations[J].The ISME Journal,2014,8(8):1566-1576.  
[37] PAJARILLO E A B,CHAE J P,KIM H B,et al.Barcoded pyrosequencing-based metagenomic analysis of the faecal microbiome of three purebred pig lines after cohabitation[J].Applied Microbiology and Biotechnology,2015,99(13):5647-5656.  
[38] CAMARINHA-SILVA A,MAUSHAMMER M,WELLMANN R,et al.Host genome influence on gut microbial composition and microbial prediction of complex traits in pigs[J].Genetics,2017,206(3):1637-1644.  
[39] GULABIVALA K,NG Y L.Endodontics[M].4th ed.Amsterdam:Elsevier Ltd.,2014.
[40] GREENING R C,LEEDLE J A Z.Enrichment and isolation of Acetitomaculum ruminis,gen.nov.,sp.nov.:acetogenic bacteria from the bovine rumen[J].Archives of Microbiology,1989,151(5):399-406.  
[41] SHKOPOROV A N,EFIMOV B A,KONDOVA I,et al.Peptococcus simiae sp.nov.isolated from rhesus macaque faeces and emended description of the genus Peptococcus[J].International Journal of Systematic and Evolutionary Microbiology,2016,66(12):5187-5191.  
[42] REYER H,OSTER M,MCCORMACK U M,et al.Host-microbiota interactions in ileum and caecum of pigs divergent in feed efficiency contribute to nutrient utilization[J].Microorganisms,2020,8(4):563.
[43] MOROTOMI M,NAGAI F,SAKON H,et al.Paraprevotella clara gen.nov.,sp.nov.and Paraprevotella xylaniphila sp.nov.,members of the family "Prevotellaceae" isolated from human faeces[J].International Journal of Systematic and Evolutionary Microbiology,2009,59(Pt 8):1895-1900.
[44] DEL DOT T,OSAWA R,STACKEBRANDT E.Phascolarctobacterium faecium gen.nov,spec.nov.,a novel taxon of the Sporomusa group of bacteria[J].Systematic and Applied Microbiology,1993,16(3):380-384.  
[45] DONALDSON G P,LEE S M,MAZMANIAN S K.Gut biogeography of the bacterial microbiota[J].Nature Reviews Microbiology,2016,14(1):20-32.  
[46] LI F Y,GUAN L L.Metatranscriptomic profiling reveals linkages between the active rumen microbiome and feed efficiency in beef cattle[J].Applied and Environmental Microbiology,2017,83(9):e00061-17.
[47] ROGOSA M.Acidaminococcus gen. n.,Acidaminococcus fermentans sp.n.,anaerobic gram-negative diplococci using amino acids as the sole energy source for growth[J].Journal of Bacteriology,1969,98(2):756-766.  
[48] MARTÍNEZ-CUESTA M C,PELÁEZ C,REQUENA T.Laboratory simulators of the colon microbiome[M]//FAINTUCH J,FAINTUCH S.Microbiome and metabolome in diagnosis,therapy,and other strategic applications.Amsterdam:Elsevier Inc.,2019.
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