导盲犬和淘汰犬肠道菌群的差异性研究
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摘要
目的探究导盲犬和淘汰犬之间肠道菌群的差异,分析两者特征性差异菌群。方法收集16只导盲犬和10只淘汰犬的新鲜粪便样本,按照犬品种和性别分为拉布拉多雄性导盲犬(GD-ML)和淘汰犬(ED-ML)、拉布拉多雌性导盲犬(GD-FL)和淘汰犬(ED-FL)、金毛雄性导盲犬(GD-MG)和淘汰犬(ED-MG),另外从中随机抽取9只导盲犬和4只淘汰犬分为导盲犬组(GD)和淘汰犬组(ED),共分为4组配对。提取粪便样本总DNA,应用PCR-DGGE技术获得肠道菌群图谱,应用相关软件和统计学方法对每组配对进行差异性分析。结果聚类分析结果显示,GD和ED,GD-ML和ED-ML,GD-FL、ED-FL和GD-MG均各归为一类,表明导盲犬和淘汰犬之间肠道菌群存在差异。肠道菌群多样性、丰富度和均匀度分析结果显示,每两个配对组之间多样性指数、丰富度指数和均匀度指数存在差异,但差异均无统计学意义。差异性条带测序分析结果显示:与ED相比,GD Megamonas funiformis YIT 11815菌株增多;与ED-ML相比,GD-ML Succinatimonas hippei YIT 12066、Lactobacillus vaginalis DSM 5837、Lactobacillus acidophilus NCFM、Faecalibacterium prausnitzii A2-165菌株增多,Collinsella aerofaciens ATCC 25986和Prevotella copri DSM 18205菌株减少;与ED-FL相比,GD-FL Ruminococcus gnavus AGR2154、Fusobacterium russii ATCC 25533菌株减少;与ED-MG相比,GD-MG Tropheryma whipplei str. Twist菌株增多。结论本研究发现导盲犬和淘汰犬肠道菌群存在差异,未来对差异菌群的进一步研究可能有助于辅助导盲犬的早期筛选。
Objective To explore the differences of gut microbiota between Guide Dogs and Eliminated Dogs, and analyze the characteristic differential gut microbiota of two groups. Method Fresh feces from 16 Guide Dogs and 10 Eliminated Dogs were collected. On the one hand, according to dogs' breed and gender, dogs were divided into Male Labrador retriever Guide Dog(GD-ML) and Eliminated Dog(ED-ML), Female Labrador retriever Guide Dog(GD-FL) and Eliminated Dog(ED-FL), Male Golden retriever Guide Dog(GD-MG) and Eliminated Dog(ED-MG).Nine Guide Dogs and 4 Eliminated Dogs randomly selected from all dogs were divided into Guide Dog(GD) and Eliminated Dog(ED). All dogs were totally divided into 4 matching groups. The genomic DNA was extracted from the fresh feces. The fingerprints of the gut microbiota were obtained by PCR-DGGE. The differences were analyzed by relevant software and statistical method. Result The result of cluster analysis showed that GD and ED, GD-ML and ED-ML, GD-FL, ED-FL and GD-MG were all classified into one class, indicating that there were differences of gut microbiota between Guide Dogs and Eliminated Dogs. There were also differences in diversity index, richness index and evenness index between each two paired groups, but the differences were not statistically significant. The result of differential strip sequencing analysis showed that compared with ED, Megamonas funiformis YIT 11815 were increased in GD; compared with ED-ML, Succinatimonas hippei YIT 12066, Lactobacillus vaginalis DSM 5837, Lactobacillus acidophilus NCFM, Faecalibacterium prausnitzii A2-165 were increased and Collinsella aerofaciens ATCC 25986, Prevotella copri DSM 18205 were decreased in GD-ML; compared with ED-FL, Ruminococcus gnavus AGR2154, Fusobacterium russii ATCC 25533 were decreased in GD-FL; compared with ED-MG, Tropheryma whipplei str. Twist were increased in GD-MG. Conclusion This study found that there were differences of gut microbiota between Guide Dogs and Eliminated Dogs. Further research on differential microbiota in the future may help to assist early screening of Guide Dogs.
Objective To explore the differences of gut microbiota between Guide Dogs and Eliminated Dogs, and analyze the characteristic differential gut microbiota of two groups. Method Fresh feces from 16 Guide Dogs and 10 Eliminated Dogs were collected. On the one hand, according to dogs' breed and gender, dogs were divided into Male Labrador retriever Guide Dog(GD-ML) and Eliminated Dog(ED-ML), Female Labrador retriever Guide Dog(GD-FL) and Eliminated Dog(ED-FL), Male Golden retriever Guide Dog(GD-MG) and Eliminated Dog(ED-MG).Nine Guide Dogs and 4 Eliminated Dogs randomly selected from all dogs were divided into Guide Dog(GD) and Eliminated Dog(ED). All dogs were totally divided into 4 matching groups. The genomic DNA was extracted from the fresh feces. The fingerprints of the gut microbiota were obtained by PCR-DGGE. The differences were analyzed by relevant software and statistical method. Result The result of cluster analysis showed that GD and ED, GD-ML and ED-ML, GD-FL, ED-FL and GD-MG were all classified into one class, indicating that there were differences of gut microbiota between Guide Dogs and Eliminated Dogs. There were also differences in diversity index, richness index and evenness index between each two paired groups, but the differences were not statistically significant. The result of differential strip sequencing analysis showed that compared with ED, Megamonas funiformis YIT 11815 were increased in GD; compared with ED-ML, Succinatimonas hippei YIT 12066, Lactobacillus vaginalis DSM 5837, Lactobacillus acidophilus NCFM, Faecalibacterium prausnitzii A2-165 were increased and Collinsella aerofaciens ATCC 25986, Prevotella copri DSM 18205 were decreased in GD-ML; compared with ED-FL, Ruminococcus gnavus AGR2154, Fusobacterium russii ATCC 25533 were decreased in GD-FL; compared with ED-MG, Tropheryma whipplei str. Twist were increased in GD-MG. Conclusion This study found that there were differences of gut microbiota between Guide Dogs and Eliminated Dogs. Further research on differential microbiota in the future may help to assist early screening of Guide Dogs.
引文
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[11] 杨絮,唐岚,单伟光.三生合欢饮抗抑郁作用及其对肠道菌群影响的实验研究[J].浙江中西医结合杂志,2017,27(7):554-557.
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[16] Kang D W,Ilhan Z E,Isern N G,et al.Differences in fecal microbial metabolites and microbiota of children with autism spectrum disorders[J].Anaerobe,2017,49:121-131.
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[21] Ballenger J C.Overview of different pharmacotherapies for attaining remission in generalized anxiety disorder[J].J Clin Psychiatry,2001,62 (Suppl 19):11-19.
[22] Jiang H Y,Zhang X,Yu Z H,et al.Altered gut microbiota profile in patients with generalized anxiety disorder[J].Journal of Psychiatric Research,2018,104:130-136.
[23] Aitbelgnaoui A,Colom A,Braniste V,et al.Probiotic gut effect prevents the chronic psychological stress-induced brain activity abnormality in mice[J].Neurogastroenterology & Motility,2014,26(4):510-520.
[24] Desbonnet L,Garrett L,Clarke G,et al.Effects of the probiotic Bifidobacterium infantis in the maternal separation model of depression[J].Neuroscience,2010,170(4):1179-1188.
[25] Gareau M G,Jury J,MacQueen G,et al.Probiotic treatment of rat pups normalises corticosterone release and ameliorates colonic dysfunction induced by maternal separation [J].Gut,2007,56(11):1522-1528.
[26] Qin N,Li D,Yang R.Next-generation sequencing technologies and the application in microbiology-a review[J].Acta Microbiologica Sinica,2011,51(4):445-457.
[27] 郑艺,张家超,郭壮,等.基于高通量测序技术分析肠道菌群及其影响因素的研究进展[J].中国食品报,2014,14(11):157-164.
[2] 王福金,王爱国,王亮,等.偏侧性测试在预测成功导盲犬中的应用[J].实验动物科学,2012,29(3):57-60.
[3] 袁野,唐芳索.导盲犬的选择及其训练方法[J].中国工作犬业,2008,(11):23-25.
[4] 王爱国,王福金,王亮,等.气质测试在导盲犬培育中的应用[J].畜牧与兽医,2012,44(2):45-47.
[5] Yano J M,Yu K,Donaldson G P,et al.Indigenous bacteria from the gut microbiota regulate host serotonin biosynthesis[J].Cell,2015,161(2):264-276.
[6] Carabotti M,Scirocco A,Maselli M A,et al.The gut-brain axis:interactions between enteric microbiota,central and enteric nervous systems[J].Annals of Gastroenterology Quarterly Publication of the Hellenic Society of Gastroenterology,2015,28(2):203-209.
[7] Jang S E,Lim S M,Jeong J J,et al.Gastrointestinal inflammation by gut microbiota disturbance induces memory impairment in mice[J].Mucosal Immunology,2017,11(2):369-379.
[8] Crumeyrolle-Arias M,Jaglin M,Bruneau A,et al.Absence of the gut microbiota enhances anxiety-like behavior and neuroendocrine response to acute stress in rats[J].Psychoneuroendocrinology,2014,42(2):207-217.
[9] Golubeva A V,Joyce S A,Moloney G,et al.Microbiota-related Changes in Bile Acid & Tryptophan Metabolism are Associated with Gastrointestinal Dysfunction in a Mouse Model of Autism[J].Ebiomedicine,2017,24(C):166-178.
[10] 范文涛,闫咏梅,别玉龙,等.大鼠卒中后肠道菌群紊乱与抑郁的关系[J].中国脑血管病杂志,2016,13(12):644-649.
[11] 杨絮,唐岚,单伟光.三生合欢饮抗抑郁作用及其对肠道菌群影响的实验研究[J].浙江中西医结合杂志,2017,27(7):554-557.
[12] Lyte M.Microbial Endocrinology in the Microbiome-Gut-Brain Axis:How Bacterial Production and Utilization of Neurochemicals Influence Behavior[J].Plos Pathogens,2013,9(11):e1003726.
[13] Dantzer R,Konsman J P,Bluthé R M,et al.Neural and humoral pathways of communication from the immune system to the brain:parallel or convergent?[J].Autonomic Neuroscience Basic & Clinical,2000,85(1):60-65.
[14] Schroeder F A,Lin C L,Crusio W E,et al.Antidepressant-like effects of the histone deacetylase inhibitor,sodium butyrate,in the mouse[J].Biological Psychiatry,2007,62(1):55-64.
[15] Li Q,Zhou J M.The microbiota-gut-brain axis and its potential therapeutic role in autism spectrum disorder[J].Neuroscience,2016,(324):131-139.
[16] Kang D W,Ilhan Z E,Isern N G,et al.Differences in fecal microbial metabolites and microbiota of children with autism spectrum disorders[J].Anaerobe,2017,49:121-131.
[17] Jiang Y,Lü X,Man C,et al.Lactobacillus acidophilus induces cytokine and chemokine production via NF-κB and p38 mitogen-activated protein kinase signaling pathways in intestinal epithelial cells[J].Clinical & Vaccine Immunology,2012,19(4):603-608.
[18] Rossi O,Berkel L A V,Chain F,et al.Faecalibacterium prausnitzii A2-165 has a high capacity to induce IL-10 in human and murine dendritic cells and modulates T cell responses[J].Scientific Reports,2016,6:18507.
[19] Sundin J,Rangel I,Repsilber D,et al.Cytokine Response after Stimulation with Key Commensal Bacteria Differ in Post-Infectious Irritable Bowel Syndrome (PI-IBS) Patients Compared to Healthy Controls[J].Plos One,2015,10(9):e0134836.
[20] Chang S E,Mishima Y,Bo L,et al.315 Induction of Bacterial Antigen-Specific Colitis by a Simplified Human Microbiota Consortium in Gnotobiotic IL-10-/-Mice[J].Infection & Immunity,2014,144(5):2239-2246.
[21] Ballenger J C.Overview of different pharmacotherapies for attaining remission in generalized anxiety disorder[J].J Clin Psychiatry,2001,62 (Suppl 19):11-19.
[22] Jiang H Y,Zhang X,Yu Z H,et al.Altered gut microbiota profile in patients with generalized anxiety disorder[J].Journal of Psychiatric Research,2018,104:130-136.
[23] Aitbelgnaoui A,Colom A,Braniste V,et al.Probiotic gut effect prevents the chronic psychological stress-induced brain activity abnormality in mice[J].Neurogastroenterology & Motility,2014,26(4):510-520.
[24] Desbonnet L,Garrett L,Clarke G,et al.Effects of the probiotic Bifidobacterium infantis in the maternal separation model of depression[J].Neuroscience,2010,170(4):1179-1188.
[25] Gareau M G,Jury J,MacQueen G,et al.Probiotic treatment of rat pups normalises corticosterone release and ameliorates colonic dysfunction induced by maternal separation [J].Gut,2007,56(11):1522-1528.
[26] Qin N,Li D,Yang R.Next-generation sequencing technologies and the application in microbiology-a review[J].Acta Microbiologica Sinica,2011,51(4):445-457.
[27] 郑艺,张家超,郭壮,等.基于高通量测序技术分析肠道菌群及其影响因素的研究进展[J].中国食品报,2014,14(11):157-164.
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