亚剂量硫酸新霉素对黄羽肉鸡回肠黏膜菌群组成的影响

doi:10.11843/j.issn.0366-6964.2016.11.019

摘要/Abstract

摘要：

以1日龄岭南黄肉鸡（♂）为试验动物，研究在日粮中长期添加亚剂量硫酸新霉素对黄羽肉鸡回肠黏膜菌群动态变化与组成的影响。于1、7、14、21、28、35及42日龄分别从A组（饲喂基础日粮）和D组（饲喂基础日粮+50 mg•kg^-1硫酸新霉素）采集鸡回肠肠段并提取黏膜细菌基因组DNA，采用PCR-DGGE技术分析鸡回肠黏膜菌群结构，同时应用16S rDNA基因序列技术，建立42日龄鸡回肠黏膜细菌16S rDNA的随机克隆文库。DGGE图谱分析发现，A、D两组1～28日龄回肠黏膜细菌结构存在明显差异，其相似性为40%～60%；A、D两组肉鸡1～35日龄菌群结构发生变化的程度不同，A组的动态变化值（%change）为9%～33%，D组的动态变化值为17%～45%。分析回肠文库发现，A组文库有94个序列，共产生9个OTUs（Operational Taxonomic Units），其中未知菌属（微杆菌属）所占克隆总数比例最大为48.94%（46/94），其次是乳杆菌属占38.30%（36/94）；D组文库有93个序列，共产生6个OTUs，其中乳杆菌属所占克隆总数比例最大为49.46%（46/93），其次是微杆菌属占45.16%（42/93）；两组回肠文库中其他菌属的组成种类与数量存在差异，且未发现与大肠杆菌相关的序列。A组与D组42日龄肉鸡回肠黏膜菌群的组成及多样性存在差异，尤其是乳杆菌属的组成比例，这种差异可能是抗生素持续压力的影响所致。

Abstract:

The experiment was conducted to study on the effects of subclinical doses of neomycin sulfate on the ileal mucosa-associated microbiota (MAM) composition and dynamic change of broiler chickens at 1 to 42 days old.Chicken ileal segments were collected in group A (basal diet) and B (basal diet with 50 mg•kg^-1 Neomycin Sulfate)，and MAM DNA were extracted at day 1，7，14，21，28，35 and 42，respectively.The community of the ileal MAM between two groups were analyzed by Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE)at various ages，and the random clone library of 16S rDNA V3 region of the MAM in ileum at 42 days old between Group A and D was established by analysis of 16S rDNA gene sequences.DGGE profiles showed that the structure of the MAM in ileum between the two groups were of remarkable difference during day 1-28，and the similarity of the MAM in ileum was 40%-60%.The alterations of MAM of ileum were remarkably different between the two groups during day 1-35，with 9%-33% change in group A and 17%-45% change in group D.Analysis of the library in ileum indicated that nine operational taxonomic units (OTUs) (form 94 gene sequences or clones) were detected in the group A，as compared with 6 OTUs (form 93 clones) in the group D.Exiguobacterium and Lactobacillus were predominant in the group A and D，counting for 48.94%，38.30% and 45.16%，49.46%， respectively.The amount of other species found in ileum in group A were different from that in group D.In addition，sequence known being related to Colibacillus were not detected in group A and D.These results indicated that the composition and diversity of the MAM in ileum at day 42 does exist difference between group A and D，especially the proportion of Lactobacillus，these difference might be caused by the continuous antibiotic pressure selection.

中图分类号:

S852.6

容庭，刘志昌，王刚，李书宏，张洁，蔡追，陈庄. 亚剂量硫酸新霉素对黄羽肉鸡回肠黏膜菌群组成的影响[J]. 畜牧兽医学报, 2016, 47(11): 2301-2309.

RONG Ting，LIU Zhi-chang，WANG Gang，LI Shu-hong，ZHANG Jie，CAI Zhui，CHEN Zhuang. Impact of Subclinical Doses of Neomycin Sulfate on the Ileal Mucosa-Associated Microbiota Composition of Broiler Chickens[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(11): 2301-2309.

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.0366-6964.2016.11.019

https://www.xmsyxb.com/CN/Y2016/V47/I11/2301

参考文献

［1］WASKMAN S A，LECHEVALIER H A.Neomycin，a new antibiotic active against streptomycin-resistant bacteria，including tuberculosis organisms［J］.Science，1949，109(2830)：305-307.
［2］CLAROT I，REGAZZETI A，AUZEIL N，et al.Analysis of neomycin sulfate and framycetin sulfate by high-performance liquid chromatography using evaporative light scattering detection［J］.J Chromatogra A，2005，1087：236-244.
［3］THONGSRISOMBOON P，LIAWRUANGRATH B，LIAWRUANGRATH S，et al.Flow injection chemiluminescence determination of Neomycin in pharmaceutical formulations［J］.J Flow Injection Anal，2010，27(1)：36-41.
［4］ISOHERRANEN N，SOBACK S.Determination of gentamicins C1，C1a，and C2 in plasma and urine by HPLC［J］.Clin Chem，2000，46(6)：837-842.
［5］HUYGHEBAERT G，DUCATELLE R，IMMERSEEL F V.An update on alternatives to antimicrobial growth promoters for broilers［J］.Vet J，2011，187(2)：182-188.
［6］DIBNER J J，RICHARDS J D.Antibiotic growth promotersin agaculture：history and mode of action［J］.Poult Sci，2005，84(4)：634-643.
［7］LI M，GONG J H，COTTRILL M，et al.Evaluation of QIAampR@ DNA Stool Mini Kit for ecological studies of gut microbiota［J］.J Microbiol Meth，2002，54(1)：13-20.
［8］张洁，容庭，刘志昌，等.两纯系鸡胃肠道黏膜细菌l6S rDNA的PCR-DGGE分析比较［J］.畜牧兽医学报，2013，44（2）：204-210.
ZHANG J，RONG T，LIU Z C，et al.Comparative analysis of 16S rDNA of the gastrointestinal mucosa-associated microbiota between two chicken pure lines by PCR-DGGE［J］.Acta Veterinaria et Zootechnica Sinica，2013，44(2)：204-210.(in Chinese)
［9］容庭，张洁，王刚，等.16S rDNA克隆文库方法分析两纯系鸡回肠及盲肠细菌的组成［J］.畜牧兽医学报，2014，45（8）：1218-1227.
RONG T， ZHANG J， WANG G， et al. Analysis of composition of the mucosa-associated microbiota in ileum and cecum between two chicken pure lines by 16S rDNA clone library［J］.Acta Veterinaria et Zootechnica Sinica, 2014,45(8):1218-1227.(in Chinese)
［10］MUYZER G，DE WAAL E C，UITTERLINDEN A G.Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S Rrna［J］.Appl Environ Microbiol，1993，59(3)：695-700.
［11］MARZORATI M，WITTEBOLLE L，BOON N，et al.How to get more out of molecular fingerprints：practical tools for microbial ecology［J］.Environ Microbiol，2008，10(6)，1571-1581.
［12］GOOD I L.The population frequencies of species and the estimation of population parameters［J］.Biometrika，1953，40：237-264.
［13］ELISHAHED M S，SENKO J M，NAJAR F Z，et al.Bacterial diversity and sulfur cycling in a mesophilic sulfide-rich spring［J］.Appl Environ Microbiol，2003，69(9)：5609-5621.
［14］HILL T C J，WALSH K A，HARRIS J A，et al.Using ecological diversity measures with bacterial communities［J］.FEMS Microbiol Ecol，2003，43(1)：1-11.
［15］SAVAGE D C.Microbial ecology of the gastrointestinal tract［J］.Annu Rev Microbiol，1977，31(70)：107-133.
［16］ZOETENDAL E G，COLLIER C T，KOIKE S，et al.Molecular ecological analysis of the gastrointestinal microbiota：A review［J］.J Nutr，2004，134(2)：465-472.
［17］ZHU X Y，ZHONG T，PANDYA Y，et al.16SrRNA-based analysis of microbiota from the cecum of broiler chickens［J］.Appl Environ Microbiol，2002，68(1)：124-137.
［18］ZOETENDAL E G，VON WRIGHT A，VILPPONEN-SALMELA T，et al.Mucosa-associated bacteria in the human gastrointestinal tract are uniformly distributed along the colon and differ from the community recovered from feces［J］.Appl Environ Microbiol， 2002，68(7)：3401-3407.
［19］LEPAGE P，SEKSIK P，SUTREN M，et al.Biodiversity of the mucosa-associated microbiota is stable along the distaldigestive tract in healthy individuals and patients with IBD［J］. Inflamm Bowel Dis， 2005，11(5)：473-480.
［20］KNARREBORG A，SIMON M A，ENGBERG RMJENSEN B B，et al.Effects of dietary fat source and subtherapeutic levels of antibiotic on the bacterial community in the ileum of broiler chickens at various ages［J］.Appl Environ Microbiol，2002，68(12)：5918-5924.
［21］SMIRNOV A，PEREZ R，AMIT-ROMACH E，et al.Mucin dynamics and microbial populations in chicken small intestine are changed by dietary probiotic and antibiotic growth promoter supplementation［J］.J Nutr，2005，135(2)：187-192.
［22］PEDROSO A A，MENTEN J F M，LAMBAIS M R，et al.Intestinal bacterial community and growth performance of chickens fed diets containing antibiotics［J］.Poult Sci，2006，85(4)：747-752.
［23］DUMONCEAUX T J，HILL J E，SEAN M，et al.Characterization of intestinal microbiota and response to dietary virginiamycin supplementation in the broiler chicken［J］.Appl Environ Microbiol，2006，72(4)：2815-2823.
［24］ZHOU H，GONG J，BRISBIN J T，et al.Appropriate chicken sample size for identifying the composition of broiler intestinal microbiota affected by dietary antibiotics，using the Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis Technique［J］. Poult Sci，2007，86(12)：2541-2549.
［25］JOHANSEN C H，BJERRUM L, PEDERSEN K.Impact of salinomycin on the intestinal microflora of broiler chickens［J］.Acta Vet Scand，2007，49(1)：30.
［26］VAN HEMERT S，HOEKMAN A J，SMITS M A，et al.Gene expression response to a Salmonella infection in the chicken intestine differ between lines［J］.Vet Immunol Immunopathol， 2006，114(3-4)：247-258.
［27］BJERRUM L，PEDERSEN K，ENGBERG R M.The influence of whole wheat feeding on Salmonella infection and gut flora composition in broilers［J］.Avian Dis，2005，49(1)：9-15.
［28］LUMPKINGS B S，BATAL A B，LEET M D.Evaluation of the bacterial community and intestinal development of different genetic lines of chickens［J］.Poult Sci，2010，89(8)：1614-1621.
［29］GONG J H，FORSTER R J，YU H，et al.Molecular analysis of bacterial populations in the ileum of broiler chickens and comparison with bacteria in the cecum［J］.FEMS Microbiol Ecol，2002，41(3)：171-179.
［30］LEY R E，BÄCKHED F，TURNBAUGH P，et al.Obesity alters gut microbial ecology［J］.Proc Natl Acad Sci U S A，2005，102(31)：11070-11075.

[1]	陈姝宇, 朱雪蛟, 周金柱, 陶然, 张雪寒, 索朗斯珠, 贡嘎, 李彬. 鸟苷酸结合蛋白GBP1和GBP2抑制猪轮状病毒体外复制[J]. 畜牧兽医学报, 2024, 55(1): 245-257.
[2]	宋雯妍, 张瀚文, 吴澳迪, 张丽燕, 刘照, 叶桐桐, 陈创夫, 盛金良. 猪繁殖与呼吸综合征病毒GP5蛋白纳米抗体的筛选及其对病毒复制的抑制效应[J]. 畜牧兽医学报, 2024, 55(1): 258-270.
[3]	谢青云, 邢蕙萱, 于岩飞, 袁厅, 熊祺琰, 熊富强, 冯志新. 猪肺炎支原体解螺旋酶RuvA的原核表达、多克隆抗体制备及活性鉴定[J]. 畜牧兽医学报, 2024, 55(1): 271-281.
[4]	陈启伟, 权衡, 陈胜利, 刘东慧, 于永峰, 李彩玉, 宫晓炜, 储岳峰. 牛支原体能量耦合因子转运蛋白S组件MbBioY摄取外源生物素的功能分析[J]. 畜牧兽医学报, 2024, 55(1): 282-289.
[5]	陈玥彤, 刘晓涵, 王芷洋, 赵宇馨, 周铁忠, 胡增金, 朱悦, 王少辉, 田明星, 丁思羽, 祁晶晶, 于圣青. 广东规模化鸡场死鸡胚中鸡毒支原体的分离鉴定、致病性及药物敏感性[J]. 畜牧兽医学报, 2024, 55(1): 290-299.
[6]	刘传霞, 王晓, 李雪雯, 鲍苗菲, 李婷婷, 陈欣, 翁长江, 郑君. 非洲猪瘟病毒pE120R蛋白单克隆抗体的制备[J]. 畜牧兽医学报, 2024, 55(1): 388-394.
[7]	杨恒, 李占鸿, 宋子昂, 高林, 李卓然, 廖德芳, 肖雷, 李华春. 帕利亚姆病毒实时荧光定量RT-PCR检测方法的建立与应用[J]. 畜牧兽医学报, 2024, 55(1): 395-400.
[8]	王莹, 李佳康, 曾悦, 史开拓, 韦丽婷, 彭佳佳, 李秋燕, 曹龙龙, 周登元. 猫杯状病毒单克隆抗体的制备[J]. 畜牧兽医学报, 2024, 55(1): 401-405.
[9]	李惠惠, 董可儿, 刘馨博, 张春晓, 马超, 陈利苹, 钟旗, 姚刚, 马雪连. 牛诺瓦病毒和牛轮状病毒双重RAA-LFD快速检测方法的建立及应用[J]. 畜牧兽医学报, 2024, 55(1): 406-412.
[10]	周建浩, 王东方, 刘影, 王淑娟, 马震原, 谢彩华, 赵雪丽, 杨海波, 冯桂丹, 康台生, 胡煜锋, 李博文, 闫若潜. 猪流行性腹泻病毒微滴式数字PCR定量检测方法的建立及初步应用[J]. 畜牧兽医学报, 2024, 55(1): 413-418.
[11]	吕若一, 司晓慧, 孙志刚, 史晓敏, 刘晓晔. 猪链球菌耐药现状分析及感染防控措施[J]. 畜牧兽医学报, 2023, 54(12): 4920-4933.
[12]	支岩, 梅晨, 刘珍邑, 乌云格日乐, 王宏俊, 胡格. 副鸡禽杆菌毒力因子研究进展[J]. 畜牧兽医学报, 2023, 54(12): 4934-4942.
[13]	王梦娇, 蒋倩, 马学军, 夏瑞阳, 郭雪萍, 孙磊, 钟旗, 马雪连, 姚刚. 新疆地区牛冠状病毒的分子流行病学调查[J]. 畜牧兽医学报, 2023, 54(12): 5125-5133.
[14]	范锦全, 张宇航, 唐午阳, 赵欣宇, 李丕顺, 郑晓峰. 地西他滨对猪圆环病毒2型的体外抑制作用[J]. 畜牧兽医学报, 2023, 54(12): 5134-5142.
[15]	张家祺, 贾浠宁, 周群, 宋鑫, 朱晨曦, 李格格, 张斌. 基于E2蛋白BVDV-1病毒样颗粒的构建及对小鼠的免疫效力评估[J]. 畜牧兽医学报, 2023, 54(12): 5143-5153.