细菌群落是实现厌氧氨氧化系统高效脱氮的核心,而厌氧氨氧化启动过程细菌群落多样性及其功能特征仍未被充分阐明.本研究采用升流式厌氧污泥床(UASB)反应器进行厌氧氨氧化系统启动,利用16S r RNA基因高通量测序技术并结合PICRUSt2功能预...细菌群落是实现厌氧氨氧化系统高效脱氮的核心,而厌氧氨氧化启动过程细菌群落多样性及其功能特征仍未被充分阐明.本研究采用升流式厌氧污泥床(UASB)反应器进行厌氧氨氧化系统启动,利用16S r RNA基因高通量测序技术并结合PICRUSt2功能预测分析,研究启动过程不同时间(d0、d30、d60和d90)细菌群落多样性及功能动态变化特征.结果表明,启动过程共检测到48个门、111个纲、269个目、457个科、840个属和1497个种;Candidatus_Brocadia和Candidatus_Kuenenia为检测到的厌氧氨氧化菌,且它们的相对丰度在启动过程不同时间存在显著差异(P <0.05).启动过程,细菌群落α多样性指数整体呈现显著的降低趋势(P <0.05),细菌群落结构呈现出明显的空间分异特征,且差异显著(R=0.846,P <0.01).PICRUSt2功能预测分析表明,启动过程,细菌群落具有丰富的功能多样性,一级功能层表现为有机系统和代谢方面较为活跃,二级功能层子功能基因丰度在厌氧氨氧化启动过程发生明显变化;细菌群落涉及49个参与氮素代谢的相关功能基因,且不同时间阶段参与硝化、反硝化、厌氧氨氧化、硝酸盐同化/异化还原和亚硝酸盐同化/异化还原过程的相关功能基因丰度发生明显变化.展开更多
为研究消毒剂对猪粪厌氧发酵系统的影响,选取不同浓度(质量分数0.02%、0.1%、0.5%)卫可(Virkon^(TM))消毒剂进行试验。结果表明:在高浓度Virkon^(TM)胁迫下,厌氧发酵系统出水中总氮(total nitrogen,TN)、氨氮(ammonia nitrogen,NH_(4)^(...为研究消毒剂对猪粪厌氧发酵系统的影响,选取不同浓度(质量分数0.02%、0.1%、0.5%)卫可(Virkon^(TM))消毒剂进行试验。结果表明:在高浓度Virkon^(TM)胁迫下,厌氧发酵系统出水中总氮(total nitrogen,TN)、氨氮(ammonia nitrogen,NH_(4)^(+)-N)、总磷(total phosphorus,TP)和化学需氧量(chemical oxygen demand,COD)等含量异常剧增,伴随TS和VS的降解率下降。对照组(CK组)和0.02%、0.1%和0.5%Virkon^(TM)试验组(分别记为L组、M组和H组)的最大产CH_(4)速率分别为63.20、71.63、73.10和38.17mL/g且CH_4总产量分别降低4.48%、16.58%(P<0.001)和86.33%(P<0.001)。随着暴露时间的延长,试验组的关键酶活呈先升后降趋势,其中,H组的S-α-GC、S-β-GC、S-ACP、S-NP及S-CAT等土壤酶活被显著性抑制(P<0.05)。进一步结合高通量测序发现,在整个厌氧发酵阶段,H组Ace指数、Chao指数和Shannon指数均显著性低于CK组(P<0.01)。在门水平上,第一优势菌群为厚壁菌门,其次为变形菌门,其中厚壁菌门相对丰度随Virkon^(TM)浓度的上升而降低,而变形菌门则相反;在属水平上,随着厌氧发酵时间的延长,束毛球菌属(Trichococcus)相对丰度均出现不同程度上升,由0.19%~0.39%提升至2.80%~4.20%,而H组中史密斯氏菌属(Smithella)的相对丰度受到极显著性(P<0.001)的抑制,较CK组下降91%。同时通过PICRUSt2功能预测分析发现,各试验组微生物群落COG(clusters of orthologous groups of proteins)功能组成结构差异并不明显,未知功能及氨基酸运输和代谢为主要优势功能;结合KO(KEGG orthology)结果与KEGG(kyoto encyclopedia of genes and genomes)数据库相关基因分析发现,高浓度Virkon^(TM)显著抑制微生物群落的生长代谢活性并削弱厌氧发酵系统的甲烷合成效能,另一方面,却诱导关键功能基因K00531(anfG)的表达活性发生超量级响应,较对照组(CK)激增约272倍。其表达水平的显著上调可能通过强化关键酶活性,使系统对Virkon^(TM)消毒副产物中典型氯代污染物(氯代烷烃及氯代烯烃类化合物)的降解效率获得提升,结果可为猪粪厌氧消化处理提供参考。展开更多
Pampus argenteus is a commercially valuable species in many countries.Its artificial cultivation has been impacted by intestinal flatulence,leading to increased mortality.However,the role of gut microbiota in this pro...Pampus argenteus is a commercially valuable species in many countries.Its artificial cultivation has been impacted by intestinal flatulence,leading to increased mortality.However,the role of gut microbiota in this process remains unclear.Therefore,this study utilized high-throughput sequencing of 16S rRNA genes to investigate the gut microbiota in both flatulent and healthy P.argenteus.A total of 227571 high-quality and classifiable reads,representing 312 Amplicon Sequence Variants(ASVs),were obtained.Proteobacteria emerged as the predominant phylum in all samples,with the flatulent group displaying the highest relative abundance(85.3%).Microbial diversity was significantly greater in samples from water than those from tissues.Composition analysis using Bray-Curtis distance via CPCoA revealed distinct gut microbiota compositions between the samples with flatulence and the healthy samples.Co-occurrence network analysis demonstrated that the bacterial community in the flatulent group exhibited disorder due to the high abundance of Vibrio.Functional predictive analysis(PICRUSt2)indicated that flatulence in P.argenteus was associated with impaired methane and lipid metabolism,potentially resulting in excessive methane synthesis utilizing hydrogen and acetic acid as substrates produced by Clostridium sensu stricto 1,Cetobacterium,and Peptoclostridium.This study holds significant implications for early diagnosis and prevention of intestinal flatulence through the regulation of gut microbiota.展开更多
For comprehensive insights into the influences of sulfate on performance,microbial community and metabolic pathways in the acidification phase of a two-phase anaerobic system,a laboratory-scale acidogenic bioreactor w...For comprehensive insights into the influences of sulfate on performance,microbial community and metabolic pathways in the acidification phase of a two-phase anaerobic system,a laboratory-scale acidogenic bioreactor was continuously operated to treat wastewater with elevated sulfate concentrations from 2000 to 14000 mg/L.The results showed that the acidogenic bioreactor could achieve sulfate reduction efficiency of greater than 70%for influent sulfate content less than 12000 mg/L.Increased sulfate induced the accumulation of volatile fatty acids(VPAs),especially propionate and butyrate,which was the primary negative effects to system performance under the high-sulfate environment.High-throughput sequencing coupled with PICRUSt2 uncovered that the accumulation of VFAs was triggered by the decreasing of genes encoding short-chain acyl-CoA dehydrogenase(EC:1.3.8.1),regulating the transformation of propanoyl-CoA to propenoyl-CoA and butanoyl-CoA to crotonyl-CoA of propionate and butyrate oxidation pathways,which made these two process hardly proceed.Besides,genes encoding(EC:1.3.8.1)were mainly carried by order Clostridiales.Desulfovibrio was the most abundant sulfate-reducing bacteria and identified as the primary host of dissimilatory sulfate reduction ftinctional genes.Functional analysis indicated the dissimilatory sulfate reduction process predominated under a low sulfate environment,but was not favored under the circumstance of high-sulfate.With the increase of sulfate,the assimilatory sulfate reduction process finally overwhelmed dissimilatory as the dominant sulfate reduction pathway in acidogenic bioreactor.展开更多
赤水河是茅台酒酿造用水的水源地,其环境承载能力和水质质量与该流域微生物的群落息息相关,而目前赤水河浮游细菌群落组成、功能及其与水质之间的关系研究开展较少.本研究以茅台酒厂采水点为中心,在其上中下游设置了W1~W6共6个采样点,采...赤水河是茅台酒酿造用水的水源地,其环境承载能力和水质质量与该流域微生物的群落息息相关,而目前赤水河浮游细菌群落组成、功能及其与水质之间的关系研究开展较少.本研究以茅台酒厂采水点为中心,在其上中下游设置了W1~W6共6个采样点,采用16S rDNA Miseq高通量测序技术研究了赤水河浮游细菌群落的组成及其功能.结果表明浮游细菌群落主要由55门、167纲、415目、706科、1431属组成,假单胞菌属(Pseudomonas)和马赛菌属(Massilia)是相对优势种群.此外,W1和W3采样点样品与其他采样点样品相比,群落组成差异较大.冗余分析表明COD_(Mn)、COD和DO是影响群落组成的显著因素(p<0.05),其与NH_(3)⁃N、pH、TN、Novosphingobium、Stenotrophomona和Pontibacter等参数是该流域浮游细菌群落网络的重要节点.使用PICRUSt2软件对该水源地微生物群落的功能进行预测,结果显示其功能主要涉及代谢(metabolism)、环境信息处理(environmental information processing)、遗传信息处理(genetic information processing)等6类生物代谢通路和碳水化合物代谢(carbohydrate metabolism)、氨基酸代谢(amino acid metabolism)、能量代谢(energy metabolism)、辅助因子和维生素的代谢(metabolism of cofactors and vitamins)等46个子功能.本研究探明了赤水河浮游细菌群落组成和功能及其与环境因子的相互联系,丰富了赤水河地区的第一手研究资料,为改善其水域环境提供了参考.展开更多
BACKGROUND Fibroblast growth factor 21(FGF21),primarily secreted by the pancreas,liver,and adipose tissues,plays a pivotal role in regulating glucose and lipid metabolism.Acute pancreatitis(AP)is a common inflammatory...BACKGROUND Fibroblast growth factor 21(FGF21),primarily secreted by the pancreas,liver,and adipose tissues,plays a pivotal role in regulating glucose and lipid metabolism.Acute pancreatitis(AP)is a common inflammatory disease with specific clinical manifestations.Many patients with diabetes present with concurrent inflammatory symptoms.Diabetes exacerbates intestinal permeability and intestinal inflammation,thus leading to the progression to AP.Our previous study indicated that FGF21 significantly attenuated susceptibility to AP in mice.AIM To investigate the potential protective role of FGF21 against AP in diabetic mice.METHODS In the present study,a mouse model of AP was established in diabetic(db)/db diabetic mice through ceruletide injections.Thereafter,the protective effects of recombinant FGF21 protein against AP were evaluated,with an emphasis on examining serum amylase(AMS)levels and pancreatic and intestinal inflammatory cytokines[interleukin(IL)-6,tumor necrosis factor-alpha(TNF-),and intestinal IL-1β].Additionally,the impact of this treatment on the histopathologic changes of the pancreas and small intestinal was examined to elucidate the role of FGF21 in diabetic mice with AP.An antibiotic(Abx)cocktail was administered in combination with FGF21 therapy to investigate whether the effect of FGF21 on AP in diabetic mice with AP was mediated through the modulation of the gut microbiota. Subsequently, thePhylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt), a bioinformaticssoftware package, was used to predict different pathways between the groups and to explore the potentialmechanisms by which the gut microbiota influenced the protective effect of FGF21.RESULTSThe results indicated that FGF21 notably diminished the levels of serum AMS (944.5 ± 15.9 vs 1732 ± 83.9, P < 0.01)and inflammatory factors including IL-6 (0.2400 ± 0.55 vs 1.233 ± 0.053, P < 0.01), TNF- (0.7067 ± 0.22 vs 1.433 ±0.051, P < 0.01), and IL-1β (1.377 ± 0.069 vs 0.3328 ± 0.02542, P < 0.01) in diabetic mice with AP. Moreover, notablesigns of recovery were observed in the pancreatic structure of the mice. The histologic evidence of inflammation inthe small intestine, including edema and villous damage, was significantly alleviated. FGF21 also significantlyaltered the composition of the gut microbiota, reestablishing the Bacteroidetes/Firmicutes ratio. Upon treatment withan Abx cocktail to deplete the gut microbiota, the FGF21 + Abx group showed lower levels of serum AMS (0.9328 ±0.075 vs 0.2249 ± 0.023, P < 0.01) and inflammatory factors (1.083 ± 0.12 vs 0.2799 ± 0.032, p < 0.01) than the FGF21group. Furthermore, the FGF21 + Abx group exhibited diminished injury to the pancreatic and small intestinaltissues, accompanied by a significant decrease in blood glucose levels (17.50 ± 1.1 vs 9.817 ± 0.69 mmol/L, P <0.001). These findings underscored the superior protective effects of the combination therapy involving an Abxcocktail with FGF21 over the FGF21 treatment alone in diabetic mice with AP. The gut microbiota compositionacross different groups was further characterized, and a differential expression analysis of gene functions wasundertaken using the PICRUSt2 prediction method. These findings suggested that FGF21 could potentially confertherapeutic effects on diabetic mice with AP by modulating the sulfate reduction I pathway and the superpathwayof n-acetylceramide degradation in the gut microbiota.CONCLUSION This study reveals the potential of FGF21 in improving pancreatic and intestinal damage recovery, reducing bloodglucose levels, and reshaping gut microbiota composition in diabetic mice with AP. Notably, the protective effectsof FGF21 are augmented when combined with the Abx cocktail.展开更多
文摘为研究消毒剂对猪粪厌氧发酵系统的影响,选取不同浓度(质量分数0.02%、0.1%、0.5%)卫可(Virkon^(TM))消毒剂进行试验。结果表明:在高浓度Virkon^(TM)胁迫下,厌氧发酵系统出水中总氮(total nitrogen,TN)、氨氮(ammonia nitrogen,NH_(4)^(+)-N)、总磷(total phosphorus,TP)和化学需氧量(chemical oxygen demand,COD)等含量异常剧增,伴随TS和VS的降解率下降。对照组(CK组)和0.02%、0.1%和0.5%Virkon^(TM)试验组(分别记为L组、M组和H组)的最大产CH_(4)速率分别为63.20、71.63、73.10和38.17mL/g且CH_4总产量分别降低4.48%、16.58%(P<0.001)和86.33%(P<0.001)。随着暴露时间的延长,试验组的关键酶活呈先升后降趋势,其中,H组的S-α-GC、S-β-GC、S-ACP、S-NP及S-CAT等土壤酶活被显著性抑制(P<0.05)。进一步结合高通量测序发现,在整个厌氧发酵阶段,H组Ace指数、Chao指数和Shannon指数均显著性低于CK组(P<0.01)。在门水平上,第一优势菌群为厚壁菌门,其次为变形菌门,其中厚壁菌门相对丰度随Virkon^(TM)浓度的上升而降低,而变形菌门则相反;在属水平上,随着厌氧发酵时间的延长,束毛球菌属(Trichococcus)相对丰度均出现不同程度上升,由0.19%~0.39%提升至2.80%~4.20%,而H组中史密斯氏菌属(Smithella)的相对丰度受到极显著性(P<0.001)的抑制,较CK组下降91%。同时通过PICRUSt2功能预测分析发现,各试验组微生物群落COG(clusters of orthologous groups of proteins)功能组成结构差异并不明显,未知功能及氨基酸运输和代谢为主要优势功能;结合KO(KEGG orthology)结果与KEGG(kyoto encyclopedia of genes and genomes)数据库相关基因分析发现,高浓度Virkon^(TM)显著抑制微生物群落的生长代谢活性并削弱厌氧发酵系统的甲烷合成效能,另一方面,却诱导关键功能基因K00531(anfG)的表达活性发生超量级响应,较对照组(CK)激增约272倍。其表达水平的显著上调可能通过强化关键酶活性,使系统对Virkon^(TM)消毒副产物中典型氯代污染物(氯代烷烃及氯代烯烃类化合物)的降解效率获得提升,结果可为猪粪厌氧消化处理提供参考。
基金funded by the National Key R&D Program of China(No.2022YFD2400100)the Ningbo 2025 Major Project of Science Technology and Innovation(No.2021Z003)+5 种基金the Public Welfare Program of Ningbo City(Science and Technology Special Commissioner Project)(No.2022S204)the National Natural Science Foundation of China(Nos.31872195,42076118 and 42306114)the China Postdoctoral Science Foundation(No.2022M721729)the General Scientific Research Projects of Zhejiang Provincial Department of Education(No.Y202249062)the Ningbo Public Welfare Science and Technology Project(No.2021S061)the Ningbo Yongjiang Talent Introduction Programme(No.2021B-029-C)。
文摘Pampus argenteus is a commercially valuable species in many countries.Its artificial cultivation has been impacted by intestinal flatulence,leading to increased mortality.However,the role of gut microbiota in this process remains unclear.Therefore,this study utilized high-throughput sequencing of 16S rRNA genes to investigate the gut microbiota in both flatulent and healthy P.argenteus.A total of 227571 high-quality and classifiable reads,representing 312 Amplicon Sequence Variants(ASVs),were obtained.Proteobacteria emerged as the predominant phylum in all samples,with the flatulent group displaying the highest relative abundance(85.3%).Microbial diversity was significantly greater in samples from water than those from tissues.Composition analysis using Bray-Curtis distance via CPCoA revealed distinct gut microbiota compositions between the samples with flatulence and the healthy samples.Co-occurrence network analysis demonstrated that the bacterial community in the flatulent group exhibited disorder due to the high abundance of Vibrio.Functional predictive analysis(PICRUSt2)indicated that flatulence in P.argenteus was associated with impaired methane and lipid metabolism,potentially resulting in excessive methane synthesis utilizing hydrogen and acetic acid as substrates produced by Clostridium sensu stricto 1,Cetobacterium,and Peptoclostridium.This study holds significant implications for early diagnosis and prevention of intestinal flatulence through the regulation of gut microbiota.
基金We gratefully acknowledge generous support provided by the National Natural Science Foundation of China(No.51978328).
文摘For comprehensive insights into the influences of sulfate on performance,microbial community and metabolic pathways in the acidification phase of a two-phase anaerobic system,a laboratory-scale acidogenic bioreactor was continuously operated to treat wastewater with elevated sulfate concentrations from 2000 to 14000 mg/L.The results showed that the acidogenic bioreactor could achieve sulfate reduction efficiency of greater than 70%for influent sulfate content less than 12000 mg/L.Increased sulfate induced the accumulation of volatile fatty acids(VPAs),especially propionate and butyrate,which was the primary negative effects to system performance under the high-sulfate environment.High-throughput sequencing coupled with PICRUSt2 uncovered that the accumulation of VFAs was triggered by the decreasing of genes encoding short-chain acyl-CoA dehydrogenase(EC:1.3.8.1),regulating the transformation of propanoyl-CoA to propenoyl-CoA and butanoyl-CoA to crotonyl-CoA of propionate and butyrate oxidation pathways,which made these two process hardly proceed.Besides,genes encoding(EC:1.3.8.1)were mainly carried by order Clostridiales.Desulfovibrio was the most abundant sulfate-reducing bacteria and identified as the primary host of dissimilatory sulfate reduction ftinctional genes.Functional analysis indicated the dissimilatory sulfate reduction process predominated under a low sulfate environment,but was not favored under the circumstance of high-sulfate.With the increase of sulfate,the assimilatory sulfate reduction process finally overwhelmed dissimilatory as the dominant sulfate reduction pathway in acidogenic bioreactor.
文摘赤水河是茅台酒酿造用水的水源地,其环境承载能力和水质质量与该流域微生物的群落息息相关,而目前赤水河浮游细菌群落组成、功能及其与水质之间的关系研究开展较少.本研究以茅台酒厂采水点为中心,在其上中下游设置了W1~W6共6个采样点,采用16S rDNA Miseq高通量测序技术研究了赤水河浮游细菌群落的组成及其功能.结果表明浮游细菌群落主要由55门、167纲、415目、706科、1431属组成,假单胞菌属(Pseudomonas)和马赛菌属(Massilia)是相对优势种群.此外,W1和W3采样点样品与其他采样点样品相比,群落组成差异较大.冗余分析表明COD_(Mn)、COD和DO是影响群落组成的显著因素(p<0.05),其与NH_(3)⁃N、pH、TN、Novosphingobium、Stenotrophomona和Pontibacter等参数是该流域浮游细菌群落网络的重要节点.使用PICRUSt2软件对该水源地微生物群落的功能进行预测,结果显示其功能主要涉及代谢(metabolism)、环境信息处理(environmental information processing)、遗传信息处理(genetic information processing)等6类生物代谢通路和碳水化合物代谢(carbohydrate metabolism)、氨基酸代谢(amino acid metabolism)、能量代谢(energy metabolism)、辅助因子和维生素的代谢(metabolism of cofactors and vitamins)等46个子功能.本研究探明了赤水河浮游细菌群落组成和功能及其与环境因子的相互联系,丰富了赤水河地区的第一手研究资料,为改善其水域环境提供了参考.
基金the 2022 Zhejiang Provincial Health Science and Technology Plan,No.2022KY1216.
文摘BACKGROUND Fibroblast growth factor 21(FGF21),primarily secreted by the pancreas,liver,and adipose tissues,plays a pivotal role in regulating glucose and lipid metabolism.Acute pancreatitis(AP)is a common inflammatory disease with specific clinical manifestations.Many patients with diabetes present with concurrent inflammatory symptoms.Diabetes exacerbates intestinal permeability and intestinal inflammation,thus leading to the progression to AP.Our previous study indicated that FGF21 significantly attenuated susceptibility to AP in mice.AIM To investigate the potential protective role of FGF21 against AP in diabetic mice.METHODS In the present study,a mouse model of AP was established in diabetic(db)/db diabetic mice through ceruletide injections.Thereafter,the protective effects of recombinant FGF21 protein against AP were evaluated,with an emphasis on examining serum amylase(AMS)levels and pancreatic and intestinal inflammatory cytokines[interleukin(IL)-6,tumor necrosis factor-alpha(TNF-),and intestinal IL-1β].Additionally,the impact of this treatment on the histopathologic changes of the pancreas and small intestinal was examined to elucidate the role of FGF21 in diabetic mice with AP.An antibiotic(Abx)cocktail was administered in combination with FGF21 therapy to investigate whether the effect of FGF21 on AP in diabetic mice with AP was mediated through the modulation of the gut microbiota. Subsequently, thePhylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt), a bioinformaticssoftware package, was used to predict different pathways between the groups and to explore the potentialmechanisms by which the gut microbiota influenced the protective effect of FGF21.RESULTSThe results indicated that FGF21 notably diminished the levels of serum AMS (944.5 ± 15.9 vs 1732 ± 83.9, P < 0.01)and inflammatory factors including IL-6 (0.2400 ± 0.55 vs 1.233 ± 0.053, P < 0.01), TNF- (0.7067 ± 0.22 vs 1.433 ±0.051, P < 0.01), and IL-1β (1.377 ± 0.069 vs 0.3328 ± 0.02542, P < 0.01) in diabetic mice with AP. Moreover, notablesigns of recovery were observed in the pancreatic structure of the mice. The histologic evidence of inflammation inthe small intestine, including edema and villous damage, was significantly alleviated. FGF21 also significantlyaltered the composition of the gut microbiota, reestablishing the Bacteroidetes/Firmicutes ratio. Upon treatment withan Abx cocktail to deplete the gut microbiota, the FGF21 + Abx group showed lower levels of serum AMS (0.9328 ±0.075 vs 0.2249 ± 0.023, P < 0.01) and inflammatory factors (1.083 ± 0.12 vs 0.2799 ± 0.032, p < 0.01) than the FGF21group. Furthermore, the FGF21 + Abx group exhibited diminished injury to the pancreatic and small intestinaltissues, accompanied by a significant decrease in blood glucose levels (17.50 ± 1.1 vs 9.817 ± 0.69 mmol/L, P <0.001). These findings underscored the superior protective effects of the combination therapy involving an Abxcocktail with FGF21 over the FGF21 treatment alone in diabetic mice with AP. The gut microbiota compositionacross different groups was further characterized, and a differential expression analysis of gene functions wasundertaken using the PICRUSt2 prediction method. These findings suggested that FGF21 could potentially confertherapeutic effects on diabetic mice with AP by modulating the sulfate reduction I pathway and the superpathwayof n-acetylceramide degradation in the gut microbiota.CONCLUSION This study reveals the potential of FGF21 in improving pancreatic and intestinal damage recovery, reducing bloodglucose levels, and reshaping gut microbiota composition in diabetic mice with AP. Notably, the protective effectsof FGF21 are augmented when combined with the Abx cocktail.
