日粮添加乳铁蛋白对断奶仔猪肠道微生物多样性的影响

doi:10.11843/j.issn.0366-6964.2023.07.025

摘要/Abstract

摘要： 旨在研究日粮添加乳铁蛋白对断奶仔猪肠道微生物多样性的影响。试验选用断奶滇太仔猪12头，初始体重为（6.12±0.54） kg，随机分为对照组、杆菌肽组和乳铁蛋白组。对照组饲喂基础日粮；杆菌肽组在基础日粮中添加0.5 g·kg^－1杆菌肽预混剂；乳铁蛋白组在基础日粮中添加150 mg·kg^－1乳铁蛋白，试验期28 d。采集试验第7、21、28天仔猪粪便，通过细菌16S rRNA基因高通量测序检测粪便微生物多样性。结果表明，各组断奶仔猪肠道微生物多样性没有显著差异（P>0.05）。在门水平上，第7、21、28天肠道优势菌为厚壁菌门和拟杆菌门，试验第7天时，各组间厚壁菌门和拟杆菌门相对丰度无显著差异（P>0.05）。与对照组相比，乳铁蛋白组厚壁菌门相对丰度降低1.43%，拟杆菌门相对丰度增加16.4%；试验第21天时，乳铁蛋白组与其余两组相比，厚壁菌门和拟杆菌门相对丰度无显著差异（P>0.05），但与对照组相比，厚壁菌门相对丰度降低16.4%，拟杆菌门相对丰度增加34.1%。试验第28天时，与对照组相比，乳铁蛋白组厚壁菌门相对丰度显著降低（P<0.05）。在属水平上，试验第7、21、28天肠道优势菌为乳杆菌属，且各组间乳杆菌属相对丰度无显著差异（P>0.05）。试验第7天时，与对照组相比，乳铁蛋白组乳杆菌属相对丰度降低了50.21％；试验第21天时，与对照组相比，乳杆菌属相对丰度降低了8.42%。试验第28天时，与对照组相比，乳铁蛋白组乳杆菌属相对丰度降低了16.09％。综上所述，本研究发现，日粮中添加乳铁蛋白不会显著影响滇太断奶仔猪的肠道微生物丰富度与多样性，断奶后7、21、28 d仔猪肠道微生物的优势菌门均为厚壁菌门与拟杆菌门，优势菌属为乳杆菌属。

关键词: 断奶仔猪, 乳铁蛋白, 肠道微生物

Abstract: This study aimed to investigate the effects of lactoferrin supplementation on gut microbial diversity in weaned piglets. Twelve weaned DianTai piglets with similar initial body weight of (6.12±0.54) kg were randomly divided into Control group, Bacitracin group and Lactoferrin group. Piglets in Control group were fed with basal diet, Bacitracin group was added with 0.5 g·kg^-1 bacitracin premix in the basal diet, while Lactoferrin group was supplemented with 150 mg·kg^-1 lactoferrin in the basal diet. The experimental period was 28 days. Fecal samples were collected on days 7, 21 and 28, and fecal microbial diversity was detected by high-throughput sequencing of bacterial 16S rRNA gene sequencing. The results showed that there was no significant difference in intestinal microbial diversity of weaned piglets (P>0.05). At the phylum level, Firmicutes and Bacteroidetes were dominant phylum on days 7, 21 and 28. At 7th day, there was no significant difference in the relative abundance of Firmicutes or Bacteroidetes among all groups (P>0.05). Compared with the control group, the relative abundance of Firmicutes in lactoferrin group decreased by 1.43% and that of Bacteroidetes increased by 16.4%. On day 21, compared with Control and Bacitracin groups, there was no significant difference in the relative abundance of Firmicutes and Bacteroidetes in Lactoferrin group (P>0.05). Compared with Control group, the relative abundance of Firmicutes decreased by 16.4% and that of Bacteroidetes increased by 34.1% in Lactoferrin group. On the 28th day of the experiment, compared with the control group, the relative abundance of Firmicutes in Lactoferrin group was significantly decreased (P<0.05). At the genus level, Lactobacillus was the dominant genus on days 7, 21 and 28, and there was no significant difference in the relative abundance of Lactobacillus among all groups (P>0.05). On day 7 of the experiment, the relative abundance of Lactobacillus in Lactoferrin group was reduced by 50.21% compared with the Control group. On day 21, the relative abundance of Lactobacillus was 8.42% lower than that of the Control group. On day 28, the relative abundance of Lactobacillus in the Lactoferrin group was 16.09% lower than that in the Control group.In conclusion, in this study, it was found that dietary lactoferrin supplementation did not significantly affect the intestinal microbial richness and diversity of weaned DianTai piglets. The dominant phylum in intestinal microbial flora of piglets at 7, 21 and 28 d after weaning were Firmicutes and Bacteroides, and the dominant genus was Lactobacillus.

Key words: weaning piglets, lactoferrin, gut microbes

中图分类号:

S828.5

纪鹏, 张斌, 张春勇, 邢笑锟, 杨佳, 刘韶娜, 方碟, 潘洪彬, 赵彦光, 安清聪. 日粮添加乳铁蛋白对断奶仔猪肠道微生物多样性的影响[J]. 畜牧兽医学报, 2023, 54(7): 2942-2955.

JI Peng, ZHANG Bin, ZHANG Chunyong, XING Xiaokun, YANG Jia, LIU Shaona, FANG Die, PAN Hongbin, ZHAO Yanguang, AN Qingcong. Effect of Dietary Supplementation of Lactoferrin on Intestinal Microbial Diversity of Weaned Piglets[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2942-2955.

参考文献 57

[1]	陈巧慧, 蓝群, 潘家华, 等.莆田黑猪肠道菌群的动态变化及其对平均日增重的影响[J].中国微生态学杂志, 2022, 34(8):900-906.CHEN Q H, LAN Q, PAN J H, et al.Dynamic distribution of gut microbiota in Putian black pigs and effect on average daily gain[J].Chinese Journal of Microecology, 2022, 34(8):900-906.(in Chinese)
[2]	RAGLAND S A, CRISS A K.From bacterial killing to immune modulation:Recent insights into the functions of lysozyme[J].PLoS Pathog, 2017, 13(9):e1006512.
[3]	QUERINJEAN P, MASSON P L, HEREMANS J F.Molecular weight, single-chain structure and amino acid composition of human lactoferrin[J].Eur J Biochem, 1971, 20(3):420-425.
[4]	CUTONE A, LEPANTO M S, ROSA L, et al.Aerosolized bovine lactoferrin counteracts infection, inflammation and iron dysbalance in a cystic fibrosis mouse model of Pseudomonas aeruginosa chronic lung infection[J].Int J Mol Sci, 2019, 20(9):2128.
[5]	BONACCORSI DI PATTI M C, CUTONE A, POLTICELLI F, et al.The ferroportin-ceruloplasmin system and the mammalian iron homeostasis machine:regulatory pathways and the role of lactoferrin[J].Biometals, 2018, 31(3):399-414.
[6]	FAN L L, YAO Q Q, WU H M, et al.Protective effects of recombinant lactoferrin with different iron saturations on enteritis injury in young mice[J].J Dairy Sci, 2022, 105(6):4791-4803.
[7]	VEGA-BAUTISTA A, DE LA GARZA M, CARRERO J C, et al.The impact of lactoferrin on the growth of intestinal inhabitant bacteria[J].Int J Mol Sci, 2019, 20(19):4707.
[8]	刘水灵, 安清聪.乳铁蛋白的免疫功能及其在养猪生产上的应用[J].家畜生态学报, 2017, 38(11):80-83.LIU S L, AN Q C.Application of lactoferrin and its immune function in swine production[J].Acta Ecologae Animalis Domastici, 2017, 38(11):80-83.(in Chinese)
[9]	NIAZ B, SAEED F, AHMED A, et al.Lactoferrin (LF):A natural antimicrobial protein[J].Int J Food Prop, 2019, 22(1):1626-1641.
[10]	LI H Y, LI P, YANG H G, et al.Investigation and comparison of the anti-tumor activities of lactoferrin, α-lactalbumin, and β-lactoglobulin in A549, HT29, HepG2, and MDA231-LM2 tumor models[J].J Dairy Sci, 2019, 102(11):9586-9597.
[11]	HERING N A, LUETTIG J, KRUG S M, et al.Lactoferrin protects against intestinal inflammation and bacteria-induced barrier dysfunction in vitro[J].Ann N Y Acad Sci, 2017, 1405(1):177-188.
[12]	SHERMAN M P.Lactoferrin and necrotizing enterocolitis[J].Clin Perinatol, 2013, 40(1):79-91.
[13]	SERRANO G, KOCHERGINA I, ALBORS A, et al.Liposomal lactoferrin as potential preventative and cure for COVID-19[J].Int J Res Health Sci, 2020, 8(1):8-15.
[14]	CHEN Y, HU S S, LI J L, et al.Bacitracin methylene disalicylate improves intestinal health by modulating its development and microbiota in weaned rabbits[J].Front Microbiol, 2021, 12:579006.
[15]	PROCTOR A, PHILLIPS G J.Differential effects of bacitracin methylene disalicylate (BMD) on the distal colon and cecal microbiota of young broiler chickens[J].Front Vet Sci, 2019, 6:114.
[16]	郭新羽, 沙玉柱, 蒲小宁, 等.环境温度对动物肠道微生物菌群影响的研究进展[J].畜牧兽医学报, 2022, 53(9):2858-2866.GUO X Y, SHA Y Z, PU X N, et al.Research progress on the effect of ambient temperature on intestinal microflora of animals[J].Acta Veterinaria et Zootechnica Sinica, 2022, 53(9):2858-2866.(in Chinese)
[17]	ZHAO W J, WANG Y P, LIU S Y, et al.The dynamic distribution of porcine microbiota across different ages and gastrointestinal tract segments[J].PLoS One, 2015, 10(2):e0117441.
[18]	NIU Q, LI P H, HAO S S, et al.Dynamic distribution of the gut microbiota and the relationship with apparent crude fiber digestibility and growth stages in pigs[J].Sci Rep, 2015, 5:9938.
[19]	HU J, NIE Y F, CHEN J W, et al.Gradual changes of gut microbiota in weaned miniature piglets[J].Front Microbiol, 2016, 7:1727.
[20]	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].Acta Veterinaria et Zootechnica Sinica, 2019, 50(9):1759-1774.(in Chinese)
[21]	赵方舟.早期乳铁蛋白干预对哺乳仔猪后肠微生物组成和肠道功能的影响[D].南京:南京农业大学, 2020.ZHAO F Z.Effects of early life lactoferrin intervention on the microbial community and intestinal function of the large intestine in suckling piglets[D].Nanjing:Nanjing Agricultural University, 2020.(in Chinese)
[22]	SCHOELER M, CAESAR R.Dietary lipids, gut microbiota and lipid metabolism[J].Rev Endocr Metab Disord, 2019, 20(4):461-472.
[23]	KARASOVA D, CRHANOVA M, BABAK V, et al.Development of piglet gut microbiota at the time of weaning influences development of postweaning diarrhea-a field study[J].Res Vet Sci, 2021, 135:59-65.
[24]	司景磊.大白猪饲料利用效率遗传和微生物标记挖掘及宿主遗传与肠道微生物互作关系的研究[D].南宁:广西大学, 2020.SI J L.Identification of the genetic and microbial biomarkers of feed efficiency trait and the interaction between host genetics and intestinal microflora in large white pigs[D].Nanning:Guangxi University, 2020.(in Chinese)
[25]	杨伟平.藏猪肠道细菌群落组成与纤维素分解菌的研究[D].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].Yangling:Northwest A&F University, 2015.(in Chinese)
[26]	KIM H B, BOREWICZ K, WHITE B A, et al.Longitudinal investigation of the age-related bacterial diversity in the feces of commercial pigs[J].Vet Microbiol, 2011, 153(1-2):124-133.
[27]	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.
[28]	THOMAS F, HEHEMANN J H, REBUFFET E, et al.Environmental and gut Bacteroidetes:the food connection[J].Front Microbiol, 2011, 2:93.
[29]	LOW K E, XING X H, MOOTE P E, et al.Combinatorial glycomic analyses to direct CAZyme discovery for the tailored degradation of canola meal non-starch dietary polysaccharides[J].Microorganisms, 2020, 8(12):1888.
[30]	POWER S E, O'TOOLE P W, STANTON C, et al.Intestinal microbiota, diet and health[J].Br J Nutr, 2014, 111(3):387-402.
[31]	FILANNINO P, DI CAGNO R, GOBBETTI M.Metabolic and functional paths of lactic acid bacteria in plant foods:Get out of the labyrinth[J].Curr Opin Biotechnol, 2018, 49:64-72.
[32]	HUANG X C, GAO J, ZHAO Y Z, et al.Dramatic remodeling of the gut microbiome around parturition and its relationship with host serum metabolic changes in sows[J].Front Microbiol, 2019, 10:2123.
[33]	YOSHII K, HOSOMI K, SAWANE K, et al.Metabolism of dietary and microbial vitamin B family in the regulation of host immunity[J].Front Nutr, 2019, 6:48.
[34]	张德明, 黄嘉訸, 李劲树, 等.猪肠道微生物及其代谢产物与肠道屏障研究进展[J].畜牧兽医学报, 2022, 53(5):1334-1344.ZHANG D M, HUANG J H, LI J S, et al.Research progress of gut microbiota, metabolites and gut barrier in pigs[J].Acta Veterinaria et Zootechnica Sinica, 2022, 53(5):1334-1344.(in Chinese)
[35]	WANG Y Z, SHAN T Z, XU Z R, 等.乳铁蛋白对断奶仔猪生长性能、肠道微生物菌群和肠形态的影响[J].何涛, 译.饲料与畜牧, 2007(7):49.WANG Y Z, SHAN T Z, XU Z R, et al.Effects of the lactoferrin (LF) on the growth performance, intestinal microflora and morphology of weanling pigs[J].HE T, trans.Feed and Husbandry, 2007(7):49.(in Chinese)
[36]	WANG Y Z, SHAN T Z, XU Z R, et al.Effects of the lactoferrin (LF) on the growth performance, intestinal microflora and morphology of weanling pigs[J].Anim Feed Sci Technol, 2007, 135(3-4):263-272.
[37]	HU W P, ZHAO J, WANG J W, et al.Transgenic milk containing recombinant human lactoferrin modulates the intestinal flora in piglets[J].Biochem Cell Biol, 2012, 90(3):485-496.
[38]	HAO Y, WANG J H, TENG D, et al.A prospective on multiple biological activities of lactoferrin contributing to piglet welfare[J].Biochem Cell Biol, 2021, 99(1):66-72.
[39]	HU P, ZHAO F Z, ZHU W Y, et al.Effects of early-life lactoferrin intervention on growth performance, small intestinal function and gut microbiota in suckling piglets[J].Food Funct, 2019, 10(9):5361-5373.
[40]	HU P, ZHAO F Z, WANG J, et al.Early-life lactoferrin intervention modulates the colonic microbiota, colonic microbial metabolites and intestinal function in suckling piglets[J].Appl Microbiol Biotechnol, 2020, 104(14):6185-6197.
[41]	DELZENNE N M, CANI P D.Interaction between obesity and the gut microbiota:relevance in nutrition[J].Annu Rev Nutr, 2011, 31:15-31.
[42]	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.
[43]	PEDERSEN R, ANDERSEN A D, HERMANN-BANK M L, et al.The effect of high-fat diet on the composition of the gut microbiota in cloned and non-cloned pigs of lean and obese phenotype[J].Gut Microbes, 2013, 4(5):371-381.
[44]	LEY R E, TURNBAUGH P J, KLEIN S, et al.Human gut microbes associated with obesity[J].Nature, 2006, 444(7122):1022-1023.
[45]	田威龙, 司景磊, 刘笑笑, 等.高脂高糖饮食对小型猪肠道微生物的影响[J].畜牧兽医学报, 2022, 53(4):1143-1153.TIAN W L, SI J L, LIU X X, et al.Effects of high-fat and high-sugar diet on intestinal microbiota in mini-pigs[J].Acta Veterinaria et Zootechnica Sinica, 2022, 53(4):1143-1153.(in Chinese)
[46]	MIN Q Q, QIN L Q, SUN Z Z, et al.Effects of metformin combined with lactoferrin on lipid accumulation and metabolism in mice fed with high-fat diet[J].Nutrients, 2018, 10(11):1628.
[47]	XIONG L, REN F Z, LV J Y, et al.Lactoferrin attenuates high-fat diet-induced hepatic steatosis and lipid metabolic dysfunctions by suppressing hepatic lipogenesis and down-regulating inflammation in C57BL/6J mice[J].Food Funct, 2018, 9(8):4328-4339.
[48]	VENKATA RAO K V, USHA RANI M, GOPALA REDDY A, et al.Amelioration of CCl4 and high fat diet-induced haematological parameters of C57BL/6 mice through lactoferrin[J].Pharma Innov J, 2022, SP-11(6):2469-2477.
[49]	LI L, MA C L, HURILEBAGEN, et al.Effects of lactoferrin on intestinal flora of metabolic disorder mice[J].BMC Microbiol, 2022, 22(1):181.
[50]	GUEVARRA R B, LEE J H, LEE S H, et al.Piglet gut microbial shifts early in life:Causes and effects[J].J Anim Sci Biotechnol, 2019, 10:1.
[51]	RIZZATTI G, LOPETUSO L R, GIBIINO G, et al.Proteobacteria:A common factor in human diseases[J].Biomed Res Int, 2017, 2017:9351507.
[52]	SHIN N R, WHON T W, BAE J W.Proteobacteria:microbial signature of dysbiosis in gut microbiota[J].Trends Biotechnol, 2015, 33(9):496-503.
[53]	BROCK J H.Lactoferrin-50 years on[J].Biochem Cell Biol, 2012, 90(3):245-251.
[54]	WEINBERG E D.The Lactobacillus anomaly:Total iron abstinence[J].Perspect Biol Med, 1997, 40(4):578-583.
[55]	GRZYWACZ K, BUTCHER J, ROMAIN G, et al.The impact of probiotics and lactoferrin supplementation on piglet gastrointestinal microbial communities[J].Biometals, 2019, 32(3):533-543.
[56]	BELLAMY W, TAKASE M, YAMAUCHI K, et al.Identification of the bactericidal domain of lactoferrin[J].Biochim Biophys Acta, 1992, 1121(1-2):130-136.
[57]	OHASHI A, MURATA E, YAMAMOTO K, et al.New functions of lactoferrin and β-casein in mammalian milk as cysteine protease inhibitors[J].Biochem Biophys Res Commun, 2003, 306(1):98-103.