AIM:To study whether selected bacterial 16S ribosomal RNA(rRNA)gene phylotypes are capable of disting- uishing irritable bowel syndrome(IBS). METHODS:The faecal microbiota of twenty volunteers with IBS,subdivided into...AIM:To study whether selected bacterial 16S ribosomal RNA(rRNA)gene phylotypes are capable of disting- uishing irritable bowel syndrome(IBS). METHODS:The faecal microbiota of twenty volunteers with IBS,subdivided into eight diarrhoea-predominant (IBS-D),eight constipation-predominant(IBS-C)and four mixed symptom-subtype(IBS-M)IBS patients,and fifteen control subjects,were analysed at three time-points with a set of fourteen quantitative real-timepolymerase chain reaction assays.All assays targeted 16S rRNA gene phylotypes putatively associated with IBS,based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobac-teria,Bacteroidetes and Firmicutes.Eight of the target phylotypes had less than 95%similarity to cultured bacterial species according to their 16S rRNA gene sequence.The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis(PCA)with linear mixed-effects models applied to the principal component scores. RESULTS:Bacterial phylotypes Clostridium cocleatum 88%,Clostridium thermosuccinogenes 85%,Coprobacillus catenaformis 91%,Ruminococcus bromii-like, Ruminococcus torques 91%,and R.torques 93%were detected from all samples analysed.A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups.The PCA on the first principal component(PC1),explaining 30.36%of the observed variation in the IBS-D patient group,was significantly altered from all other sample groups(IBS-D vs control, P=0.01;IBS-D vs IBS-M,P=0.00;IBS-D vs IBS-C, P=0.05).Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria.A phy- lotype with 85%similarity to C.thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects(-4.08±0.90 vs -3.33±1.16,P=0.04)and IBS-D and IBS-M subjects (-4.08±0.90 vs-3.08±1.38,P=0.05).Furthermore,a phylotype with 94%similarity to R.torques was more prevalent in IBS-D patients'intestinal micro- biota than in that of control subjects(-2.43±1.49 vs -4.02±1.63,P=0.01).A phylotype with 93%simi- larity to R.torques was associated with control sam- ples when compared with IBS-M(-2.41±0.53 vs -2.92±0.56,P=0.00).Additionally,a R.bromii-like phylotype was associated with IBS-C patients in com- parison to control subjects(-1.61±1.83 vs-3.69± 2.42,P=0.01).All of the above mentioned phylotype specific alterations were independent of the effect of time. CONCLUSION:Significant phylotype level alterationsin the intestinal microbiotas of IBS patients were observed,further emphasizing the possible contribution of the gastrointestinal microbiota in IBS.展开更多
Objective To analyze the characteristics of the intestinal microflora in patients with breast fibroadenoma using 16S ribosomal RNA(rRNA)high-throughput sequencing.Methods Fecal samples from 20 patients with breast fib...Objective To analyze the characteristics of the intestinal microflora in patients with breast fibroadenoma using 16S ribosomal RNA(rRNA)high-throughput sequencing.Methods Fecal samples from 20 patients with breast fibroadenoma and 36 healthy subjects were randomly collected and analyzed using high-throughput sequencing technology for 16S rRNA V4 region sequencing,and the alpha diversity(Chao index,Shannon index)was calculated using Mothur(v.1.39.5)software.Beta diversity was analyzed using QIIME(v1.80).SPSS software(version 23.0)and the t-test of two independent samples were used to analyze differences in the abundance of bacteria between the two groups.Results Compared with that in the healthy control group,theαdiversity of the intestinal microflora in breast fibroadenoma patients increased,but the difference was not statistically significant(P>0.05).At the phylum level,significant differences were observed between the two groups.The abundance of Firmicutes was higher in the breast fibroadenoma group(P<0.05),whereas the abundance of Synergistetes was higher in the healthy control group(P<0.005).A total of five bacterial genera showed significant differences between the two groups:the breast fibroadenoma group showed higher levels of Bautia(P<0.005),Coprococcus(P<0.005),Roseburia(P<0.05),and Ruminococcus(P<0.005),whereas Sutterella was more abundant in the healthy control group than in the breast fibroadenoma group(P<0.05).Conclusion The diversity and abundance of the intestinal flora in patients with breast fibroadenoma are significantly different from those in healthy subjects,suggesting that an imbalance in the intestinal flora is correlated with the occurrence of breast fibroadenoma.展开更多
The number of base pairs in the 16S rRNA secondary structures of 51 bacterial sequences was counted, and the number of hydrogen bonds was estimated. The number of hydrogen bonds was highly correlated with the optimal ...The number of base pairs in the 16S rRNA secondary structures of 51 bacterial sequences was counted, and the number of hydrogen bonds was estimated. The number of hydrogen bonds was highly correlated with the optimal growth temperature (OGT) rather than with the G + C content. Paired and unpaired nucleotides in mesophiles were compared to those in thermophiles. OGT exhibited a relationship with paired nucleotides but not with unpaired nucleotides. The total number of paired as well as unpaired nucleotides in mesophiles was very similar to that in thermophiles. However, the components in base pairs in mesophiles significantly differed from those in thermophiles. As compared with mesophiles, the number of G·C base pairs in thermophiles was high whereas that of A·U base pairs was low. In this study, we showed that hydrogen bonds are important for stabilizing 16S rRNAs at high temperatures.展开更多
Background: Passerida is the largest avian radiation within the order Passeriformes. Current understanding of the high-level relationships within Passerida is based on DNA–DNA hybridizations; however, the phylogeneti...Background: Passerida is the largest avian radiation within the order Passeriformes. Current understanding of the high-level relationships within Passerida is based on DNA–DNA hybridizations; however, the phylogenetic relationships within this assemblage have been the subject of many debates.Methods: We analyzed the 12 S ribosomal RNA gene from 49 species of Passerida, representing 14 currently recognized families, to outline the phylogenetic relationships within this group.Results: Our results identified the monophyly of the three superfamilies in Passerida: Sylvioidea, Muscicapoidea and Passeroidea. However, current delimitation of some species is at variance with our phylogeny estimate. First, the Parus major, which had been placed as a distinct clade sister to Sylvioidea was identified as a member of the super family;second, the genus Regulus was united with the Sturnidae and nested in the Muscicapoidea clade instead of being a clade of Passerida.Conclusion: Our results were consistent with Johansson's study of the three superfamilies except for the al ocation of two families, Paridae and Regulidae.展开更多
To study the phylogeny of Hynobiidae, we amplified DNA fragments of 470 bp 16S ribosomal RNA (16S rRNA) gene on mitochondrial DNA from Ranodon sibiricus and Ranodon tsinpaensis. PCR products were cloned into PMD18 T v...To study the phylogeny of Hynobiidae, we amplified DNA fragments of 470 bp 16S ribosomal RNA (16S rRNA) gene on mitochondrial DNA from Ranodon sibiricus and Ranodon tsinpaensis. PCR products were cloned into PMD18 T vector after purification. These sequences were determined and deposited in the GenBank (accession numbers: AY373459 for Ranodon sibiricus, AY372534 for Ranodon tsinpaensis). By comparing the nucleotide differences of 16S ribosomal RNA sequences among Liua shihi,Pseudohynobius flavomaculatus and Batrachuperus genus from GenBank database,we analyzed the divergences and base substitution among these sequences with the MEGA software. The molecular results support that B tibetanus, B pinchonii and B karlschmidti are classified into three valid species. Liua shihi has closer phylogenetic relationships to Ranodon tsinpaensis than to other species. More our results reveal that Pseudohynobius flavomaculatus is not a synonym of Ranodon tsinpaensis. .展开更多
将养殖粪便进行资源化处理,尤其是将粪便堆肥发酵后变为生物肥料还田,具有重要的经济、社会和生态效益。之前关于细菌在堆肥过程中的研究,大部分采用实验室培养、分离、鉴定的方法,由于受培养方式的限制,仅能分析粪肥中有限的细菌类别。...将养殖粪便进行资源化处理,尤其是将粪便堆肥发酵后变为生物肥料还田,具有重要的经济、社会和生态效益。之前关于细菌在堆肥过程中的研究,大部分采用实验室培养、分离、鉴定的方法,由于受培养方式的限制,仅能分析粪肥中有限的细菌类别。16S r RNA基因作为生物物种的特征核酸序列,被认为是最适于细菌系统发育和分类鉴定研究的指标。本研究使用16S r RNA基因高通量测序技术,分析了牛粪自然发酵与添加益生菌剂发酵过程中细菌种群的多样性变化。结果表明,1)新鲜牛粪、自然发酵1个月、自然发酵6个月的牛粪中细菌种群并没有明显的变化规律,说明自然发酵过程主要依赖于新鲜牛粪中携带的细菌种群;2)添加益生菌发酵后,细菌种群明显不同于不自然发酵过程中的细菌种群,其中变形菌门(Proteobacteria)细菌显著增加,而厚壁菌门(Firmicutes)细菌显著减少,说明益生菌剂能够显著改变堆肥过程中的细菌种群。本研究对于理解牛粪堆肥过程、提高堆肥效果,以及新型堆肥益生菌剂的开发都具有重要意义。展开更多
收集人、大猪、小猪、大鼠、小鼠以及鸡五个不同个体的粪便样品,每个个体取两个平行样,提取总DNA;PCR扩增,获得16S r RNA V6标签片段;Illumina测序;经BIPES以及QIIME分析并比较菌群结构及多样性。研究结果发现,不同物种之间肠道菌群差...收集人、大猪、小猪、大鼠、小鼠以及鸡五个不同个体的粪便样品,每个个体取两个平行样,提取总DNA;PCR扩增,获得16S r RNA V6标签片段;Illumina测序;经BIPES以及QIIME分析并比较菌群结构及多样性。研究结果发现,不同物种之间肠道菌群差异较大。五类物种肠道菌群均以厚壁菌门、拟杆菌门、以及变形菌门为主,但鸡的厚壁菌门显著减少,而变形菌门显著增加。从?多样性角度来看,猪肠道菌群种属丰富度及Shannon指数均显著高于人及鸡肠道菌群。从?多样性角度,尽管不同人之间的肠道菌群相似度较低,但不同物种之间相比较,小猪与大鼠肠道菌群与人相似性高于小鼠和鸡肠道菌群。与人类相比,小猪的肠道微生物组最相近,而鸡的肠道菌群相似度最低。展开更多
基金Supported by The Finnish Funding Agency for Technologyand Innovation,Tekes,grants No.945/401/00 and 40160/05the Finnish Graduate School of Applied Biosciences,the Academy of Finland,Grant No.214 157the Centre of Excellence on Microbial Food Safety Research,Academy of Finland
文摘AIM:To study whether selected bacterial 16S ribosomal RNA(rRNA)gene phylotypes are capable of disting- uishing irritable bowel syndrome(IBS). METHODS:The faecal microbiota of twenty volunteers with IBS,subdivided into eight diarrhoea-predominant (IBS-D),eight constipation-predominant(IBS-C)and four mixed symptom-subtype(IBS-M)IBS patients,and fifteen control subjects,were analysed at three time-points with a set of fourteen quantitative real-timepolymerase chain reaction assays.All assays targeted 16S rRNA gene phylotypes putatively associated with IBS,based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobac-teria,Bacteroidetes and Firmicutes.Eight of the target phylotypes had less than 95%similarity to cultured bacterial species according to their 16S rRNA gene sequence.The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis(PCA)with linear mixed-effects models applied to the principal component scores. RESULTS:Bacterial phylotypes Clostridium cocleatum 88%,Clostridium thermosuccinogenes 85%,Coprobacillus catenaformis 91%,Ruminococcus bromii-like, Ruminococcus torques 91%,and R.torques 93%were detected from all samples analysed.A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups.The PCA on the first principal component(PC1),explaining 30.36%of the observed variation in the IBS-D patient group,was significantly altered from all other sample groups(IBS-D vs control, P=0.01;IBS-D vs IBS-M,P=0.00;IBS-D vs IBS-C, P=0.05).Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria.A phy- lotype with 85%similarity to C.thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects(-4.08±0.90 vs -3.33±1.16,P=0.04)and IBS-D and IBS-M subjects (-4.08±0.90 vs-3.08±1.38,P=0.05).Furthermore,a phylotype with 94%similarity to R.torques was more prevalent in IBS-D patients'intestinal micro- biota than in that of control subjects(-2.43±1.49 vs -4.02±1.63,P=0.01).A phylotype with 93%simi- larity to R.torques was associated with control sam- ples when compared with IBS-M(-2.41±0.53 vs -2.92±0.56,P=0.00).Additionally,a R.bromii-like phylotype was associated with IBS-C patients in com- parison to control subjects(-1.61±1.83 vs-3.69± 2.42,P=0.01).All of the above mentioned phylotype specific alterations were independent of the effect of time. CONCLUSION:Significant phylotype level alterationsin the intestinal microbiotas of IBS patients were observed,further emphasizing the possible contribution of the gastrointestinal microbiota in IBS.
基金Supported by a grant from the Qingdao Pharmaceutical Research Guidance Plan 2019(No.2019-WJZD140).
文摘Objective To analyze the characteristics of the intestinal microflora in patients with breast fibroadenoma using 16S ribosomal RNA(rRNA)high-throughput sequencing.Methods Fecal samples from 20 patients with breast fibroadenoma and 36 healthy subjects were randomly collected and analyzed using high-throughput sequencing technology for 16S rRNA V4 region sequencing,and the alpha diversity(Chao index,Shannon index)was calculated using Mothur(v.1.39.5)software.Beta diversity was analyzed using QIIME(v1.80).SPSS software(version 23.0)and the t-test of two independent samples were used to analyze differences in the abundance of bacteria between the two groups.Results Compared with that in the healthy control group,theαdiversity of the intestinal microflora in breast fibroadenoma patients increased,but the difference was not statistically significant(P>0.05).At the phylum level,significant differences were observed between the two groups.The abundance of Firmicutes was higher in the breast fibroadenoma group(P<0.05),whereas the abundance of Synergistetes was higher in the healthy control group(P<0.005).A total of five bacterial genera showed significant differences between the two groups:the breast fibroadenoma group showed higher levels of Bautia(P<0.005),Coprococcus(P<0.005),Roseburia(P<0.05),and Ruminococcus(P<0.005),whereas Sutterella was more abundant in the healthy control group than in the breast fibroadenoma group(P<0.05).Conclusion The diversity and abundance of the intestinal flora in patients with breast fibroadenoma are significantly different from those in healthy subjects,suggesting that an imbalance in the intestinal flora is correlated with the occurrence of breast fibroadenoma.
文摘The number of base pairs in the 16S rRNA secondary structures of 51 bacterial sequences was counted, and the number of hydrogen bonds was estimated. The number of hydrogen bonds was highly correlated with the optimal growth temperature (OGT) rather than with the G + C content. Paired and unpaired nucleotides in mesophiles were compared to those in thermophiles. OGT exhibited a relationship with paired nucleotides but not with unpaired nucleotides. The total number of paired as well as unpaired nucleotides in mesophiles was very similar to that in thermophiles. However, the components in base pairs in mesophiles significantly differed from those in thermophiles. As compared with mesophiles, the number of G·C base pairs in thermophiles was high whereas that of A·U base pairs was low. In this study, we showed that hydrogen bonds are important for stabilizing 16S rRNAs at high temperatures.
基金supported by the National Natural Science Foundation of China (NSFC, 31000191, 31330073)the Postdoctoral Science Foundation of China (2011 M500537)+2 种基金the Natural Science Foundation of Hebei Province (NSFHB, 2012205018)the Natural Science Foundation of the Department of Education, Hebei Province (YQ2014024) to D. LiNSFHB (2013205018) to Y. Wu
文摘Background: Passerida is the largest avian radiation within the order Passeriformes. Current understanding of the high-level relationships within Passerida is based on DNA–DNA hybridizations; however, the phylogenetic relationships within this assemblage have been the subject of many debates.Methods: We analyzed the 12 S ribosomal RNA gene from 49 species of Passerida, representing 14 currently recognized families, to outline the phylogenetic relationships within this group.Results: Our results identified the monophyly of the three superfamilies in Passerida: Sylvioidea, Muscicapoidea and Passeroidea. However, current delimitation of some species is at variance with our phylogeny estimate. First, the Parus major, which had been placed as a distinct clade sister to Sylvioidea was identified as a member of the super family;second, the genus Regulus was united with the Sturnidae and nested in the Muscicapoidea clade instead of being a clade of Passerida.Conclusion: Our results were consistent with Johansson's study of the three superfamilies except for the al ocation of two families, Paridae and Regulidae.
文摘To study the phylogeny of Hynobiidae, we amplified DNA fragments of 470 bp 16S ribosomal RNA (16S rRNA) gene on mitochondrial DNA from Ranodon sibiricus and Ranodon tsinpaensis. PCR products were cloned into PMD18 T vector after purification. These sequences were determined and deposited in the GenBank (accession numbers: AY373459 for Ranodon sibiricus, AY372534 for Ranodon tsinpaensis). By comparing the nucleotide differences of 16S ribosomal RNA sequences among Liua shihi,Pseudohynobius flavomaculatus and Batrachuperus genus from GenBank database,we analyzed the divergences and base substitution among these sequences with the MEGA software. The molecular results support that B tibetanus, B pinchonii and B karlschmidti are classified into three valid species. Liua shihi has closer phylogenetic relationships to Ranodon tsinpaensis than to other species. More our results reveal that Pseudohynobius flavomaculatus is not a synonym of Ranodon tsinpaensis. .
文摘将养殖粪便进行资源化处理,尤其是将粪便堆肥发酵后变为生物肥料还田,具有重要的经济、社会和生态效益。之前关于细菌在堆肥过程中的研究,大部分采用实验室培养、分离、鉴定的方法,由于受培养方式的限制,仅能分析粪肥中有限的细菌类别。16S r RNA基因作为生物物种的特征核酸序列,被认为是最适于细菌系统发育和分类鉴定研究的指标。本研究使用16S r RNA基因高通量测序技术,分析了牛粪自然发酵与添加益生菌剂发酵过程中细菌种群的多样性变化。结果表明,1)新鲜牛粪、自然发酵1个月、自然发酵6个月的牛粪中细菌种群并没有明显的变化规律,说明自然发酵过程主要依赖于新鲜牛粪中携带的细菌种群;2)添加益生菌发酵后,细菌种群明显不同于不自然发酵过程中的细菌种群,其中变形菌门(Proteobacteria)细菌显著增加,而厚壁菌门(Firmicutes)细菌显著减少,说明益生菌剂能够显著改变堆肥过程中的细菌种群。本研究对于理解牛粪堆肥过程、提高堆肥效果,以及新型堆肥益生菌剂的开发都具有重要意义。
文摘收集人、大猪、小猪、大鼠、小鼠以及鸡五个不同个体的粪便样品,每个个体取两个平行样,提取总DNA;PCR扩增,获得16S r RNA V6标签片段;Illumina测序;经BIPES以及QIIME分析并比较菌群结构及多样性。研究结果发现,不同物种之间肠道菌群差异较大。五类物种肠道菌群均以厚壁菌门、拟杆菌门、以及变形菌门为主,但鸡的厚壁菌门显著减少,而变形菌门显著增加。从?多样性角度来看,猪肠道菌群种属丰富度及Shannon指数均显著高于人及鸡肠道菌群。从?多样性角度,尽管不同人之间的肠道菌群相似度较低,但不同物种之间相比较,小猪与大鼠肠道菌群与人相似性高于小鼠和鸡肠道菌群。与人类相比,小猪的肠道微生物组最相近,而鸡的肠道菌群相似度最低。
