β-Proteobacteria菌降解甲基叔丁基醚的条件及中间代谢产物研究

采用可利用甲基叔丁基醚(methyltert-butyl ether,MTBE)为唯一碳源和能源生长的1株β-Proteobacteria菌进行MTBE在密闭系统中的降解试验,确定了该菌降解MTBE的最适条件为:培养液初始pH值7.2,初始细胞浓度107cells/mL,初始MTBE浓度为25 mg/L.考察了密封培养系统内培养液溶解氧对降解效果的影响,结果表明,在培养系统密闭前充入氧气可提高菌体对MTBE的降解速率.以气相色谱-质谱联用法检测到MTBE降解主要中间代谢产物是叔丁基醇、异丙醇、丙酮.在选择离子扫描模式下定量分析,得到降解过程中主要中间代谢产物的浓度变化曲线,据此推断MTBE的降解途径属于“丙酮途径”.

Aerobic degradation of methyl tert-butyl ether by a Proteobacteria strain in a closed culture system

The contamination of methyl ten-butyl ether （MTBE） in underground waters has become a widely concerned problem all over the world. In this study, a novel dosed culture system with oxygen supplied by H2O2 was introduced for MTBE aerobic biodegradation. After 7 d, almost all MTBE was degraded by a pure culture, a member of β-Proteobacteria named as PMI, in a closed system with oxygen supply, while only 40% MTBE was degraded in one without oxygen supply. Dissolved oxygen （DO） levels of the broth in closed systems respectively with and without H2O2 were about 5-6 and 4 mg/L. Higher DO may improve the activity of monooxygemase, which is the key enzyme of metabolic pathway from MTBE to tert-butyl alcohol and finally to CO2, and may result in the increase of the degrading activity of PM1 cell. The purge and trap GC-MS result of the broth in closed systems showed that tea-butyl alcohol, isopronol and acetone were the main intermediate products.

Phylogeny of γ-proteobacteria inferred from comparisons of 3’ end 16S rRNA gene and 5’ end 16S-23S ITS nucleotide sequences

The phylogeny of γ-proteobacteria was inferred from nucleotide sequence comparisons of a short 232 nucleotide sequence marker. A total of 64 γ-proteobacterial strains from 13 Orders, 22 families, 40 genera and 59 species were analyzed. The short 232 nucleotide sequence marker used here was a combination of a 157 nucleotide sequence at the 3’ end of the 16S rRNA gene and a 75 nucleotide sequence at the 5’ end of the 16S-23S Internal Transcribed Spacer (ITS) sequence. Comparative analyses of the 3’ end of the 16S rRNA gene nucleotide sequence showed that the last 157 bp were conserved among strains from same species and less conserved in more distantly related species. This 157 bp sequence was selected as the first part in the construction of our nucleotide sequence marker. A bootstrapped neighbor-joining tree based on the alignment of this 157 bp was constructed. This 157 bp could distinguish γ-proteobacterial species from different genera from same family. Closely related species could not be distinguished. Next, an alignment of the 16S-23S ITS nucleotide sequences of alleles from same bacterial strain was performed. The first 75 bp at the 5’ end of the 16S-23S ITS was highly conserved at the intra-strain level. It was selected as the second part in the construction of our nucleotide sequence marker. Finally, a bootstrapped neighbor-joining tree based on the alignment of this 232 bp sequence was constructed. Based on the topology of the neighbour-joining tree, four major Groups, Group I to IV, were revealed with several sub-groups and clusters. Our results, based on the 232 bp sequence were, in general, in agreement with the phylogeny of γ-proteobacteria based on the 16S rRNA gene. The use of this 232 bp sequence as a phylogenetic marker presents several advantages over the use of the entire 16S rRNA gene or the generation of extensive phenotypic and genotypic data in phylogenetic analyses. First, this marker is not allele-dependant. Second, this 232 bp marker contains 157 bp from the 3’ end of the 16S rRNA gene and 75 bp from the 5’ end of the 16S-23S ITS. The 157 bp allows discrimination among distantly related species. Owing to its higher rate of nucleotide substitutions, the 75 bp adds discriminating power among closely related species from same genus and closely related genera from same family. Because of its higher percentage of nucleotide sequence divergence than the 16S rRNA gene, the 232 bp marker can better discriminate among closely related γ-proteobacterial species. Third, the method is simple, rapid, suited to large screening programs and easily accessible to most laboratories. Fourth, this marker can also reveal γ-proteobacterial species which may appear misassigned and for which additional characterization appear warranted.

Microbiomes of Top and Sub-Layers of Semi-Arid Soils in North-Eastern Nigeria Are Rich in Firmicutes and Proteobacteria with Surprisingly High Diversity of Rare Species

Borno state is the second largest state in Nigeria with over 70,000 square kilometers of diverse ecosystems including parts of the fertile Lake Chad basin. However, more than 2/3 of this landmass is threatened with drought, advancing desertification and degraded soils. Most restoration efforts involve revegetation, which in the past has met with limited success. Microbial communities of soils play a pivotal role in soil fertility and plant cover. We conducted the first metagenomic amplicon sequencing study, comparing two soil depths to determine whether soil bacteria abundance and diversity in the harsh bare soils were sufficient to sustain greening efforts. The goal was to glean insights to guide microbial inoculant formulation needed in the region. Samples from top (0 - 15 cm) and sub (16 - 65 cm) soils were collected from five strategic locations in the state. Using next generation Illumina sequencing protocols, total DNA extracted directly from the soils was sequenced and analyzed by QIIME. Metadata collected from site showed scorching temperatures of over 46?C, near zero moisture level and a pH of about 6 for top soil. At 65 cm depth, the temperature averaged 32?C with a pH of 5 and significantly higher soil moisture of 0.1%. The bacterial community structure was unexpectedly very diverse at both soil depths samples, recording a ChaO1 index ranging from 909 to 4296 and a Shannon diversity range of 3.54 to 6.33. The most abundant phyla in both soil depths were the Firmicutes and Proteobacteria;however the relative abundance of composite lower taxa was strikingly different. Operational taxonomic units and diversity indices were highest for top soils and were dominated by members of resilient groups of Actinobacteria, Firmucutes, Acidobacteria and numerous other less well-known taxa whose individual relative abundance did not exceed 3% of total population. The high diversity and richness of Proteobacteria (at 65 cm depth), some of which are key to soil fertility, suggest that revegetation efforts could be improved by shifting the gradient of these microbiota upwards using shades and micro-irrigation. Soils in semi-arid regions in Nigeria contain numerous operational taxonomic bacterial groups with potential thermophilic and drought genetic resources to be mined. Microbial community structure beneath the top soil appears stable and should be the target sample for the assessments of climatic change impact on microbial community structure in environments like this.

New insights into the interactions between the gut microbiota and the inflammatory response to ulcerative colitis in a mouse model of dextran sodium sulfate and possible mechanisms of action for treatment with PE&AFWE

Background:Inflammatory bowel disease(IBD),comprising Crohn's disease(CD)and ulcerative colitis(UC),is a heterogeneous state of chronic intestinal inflammation.Intestinal innate immunity,including innate immune cells,defends against pathogens and excessive entry of gut microbiota,while preserving immune tolerance to resident intestinal microbiota,and may be characterized by its capacity to produce a rapid and nonspecific reaction.The association between microbiota dysbiosis and the pathogenesis of IBD is complex and dynamic.When the intestinal ecosystem is in dysbiosis,the reduced abundance and diversity of intestinal gut microbiota make the host more vulnerable to the attack of exogenous and endogenous pathogenic gut microbiota.The aim of our study was to comprehensively assess the relationship between microbial populations within UC,the signaling pathways of pathogenic gut microbe therein and the inflammatory response,as well as to understand the effects of using PE&AFWE(poppy extract[Papaver nudicaule L.]and Artemisia frigida Willd.extract)on UC modulation.Methods:A UC mouse model was established by inducing SPF-grade C57BL/6 mice using dextrose sodium sulfate(DSS).Based on metagenomic sequencing to characterize the gut microbiome,the relationship between gut microbiota dysbiosis and gut microbiota was further studied using random forest and Bayesian network analysis methods,as well as histopathological analysis.Results:(1)We found that the 5 gut microbiota with the highest relative abundance of inflammatory bowel disease UC model gut microbiota were consistent with the top 5 ranked natural bacteria.There were three types of abundance changes in the model groups:increases(Chlamydiae/Proteobacteria and Deferribacteres),decreases(Firmicutes),and no significant changes(Bacteroidetes).The UC model group was significantly different from the control group,with 1308 differentially expressed species with abundance changes greater than or equal to 2-fold.(2)The proportion of the fecal flora in the UC group decreased by 37.5%in the Firmicutes and increased by 14.29%in the proportion of Proteobacteria compared to the control group before treatment.(3)The significantly enriched and increased signaling pathways screened were the'arachidonic acid metabolic pathway'and the'phagosomal pathway',which both showed a decreasing trend after drug administration.(4)Based on the causal relationship between different OTUs and the UC model/PE&AFWE administration,screening for directly relevant OTU networks,the UC group was found to directly affect OTU69,followed by a cascade of effects on OTU12,OTU121,OTU93,and OTU7,which may be the pathway of action that initiated the pathological changes in normal mice.(5)We identified a causal relationship between common differentially expressed OTUs and PE&AFWE and UC in the pre-and post-PE&AFWE-treated groups.Thereby,we learned that PE&AFWE can directly affect OTU90,after which it inhibits UC,inhibiting the activity of arachidonic acid metabolic pathway by affecting OTU118,which in turn inhibits the colonization of gut microbiota by OTU93 and OTU7.(6)Histopathological observation and scoring(HS)of the colon showed that there was a significant difference between the model group and the control group(p<0.001),and that there was a significant recovery in both the sulfasalazine(SASP)and the PE&AFWE groups after the administration of the drug(p<0.0001).Conclusion:We demonstrated causal effects and inflammatory metabolic pathways in gut microbiota dysbiosis and IBD,with five opportunistic pathogens directly contributing to IBD.PE&AFWE reduced the abundance of proteobacteria in the gut microbiota,and histopathology showed significant improvement.

Gut microbiota in preterm infants receiving breast milk or mixed feeding

BACKGROUND Preterm birth is the leading cause of mortality in newborns,with very-low-birthweight infants usually experiencing several complications.Breast milk is considered the gold standard of nutrition,especially for preterm infants with delayed gut colonization,because it contains beneficial microorganisms,such as Lactobacilli and Bifidobacteria.AIM To analyze the gut microbiota of breastfed preterm infants with a birth weight of 1500 g or less.METHODS An observational study was performed on preterm infants with up to 36.6 wk of gestation and a birth weight of 1500 g or less,born at the University Hospital Dr.JoséEleuterio González at Monterrey,Mexico.A total of 40 preterm neonates were classified into breast milk feeding(BM)and mixed feeding(MF)groups(21 in the BM group and 19 in the MF group),from October 2017 to June 2019.Fecal samples were collected before they were introduced to any feeding type.After full enteral feeding was achieved,the composition of the gut microbiota was analyzed using 16S rRNA gene sequencing.Numerical variables were compared using Student’s t-test or using the Mann–Whitney U test for nonparametric variables.Dominance,evenness,equitability,Margalef’s index,Fisher’s alpha,Chao-1 index,and Shannon’s diversity index were also calculated.RESULTS No significant differences were observed at the genus level between the groups.Class comparison indicated higher counts of Alphaproteobacteria and Betaproteobacteria in the initial compared to the final sample of the BM group(P<0.011).In addition,higher counts of Gammaproteobacteria were detected in the final than in the initial sample(P=0.040).According to the Margalef index,Fisher’s alpha,and Chao-1 index,a decrease in species richness from the initial to the final sample,regardless of the feeding type,was observed(P<0.050).The four predominant phyla were Bacteroidetes,Actinobacteria,Firmicutes,and Proteobacteria,with Proteobacteria being the most abundant.However,no significant differences were observed between the initial and final samples at the phylum level.CONCLUSION Breastfeeding is associated with a decrease in Alphaproteobacteria and Betaproteobacteria and an increase of Gammaproteobacteria,contributing to the literature of the gut microbiota structure of very low-birth-weight,preterm.

城镇污水厂尾水排放对嘉陵江流域微生物群落的潜在影响

为了探究城镇污水排放对嘉陵江流域水体中微生物群落的潜在影响,于2021年12月下旬,采集嘉陵江(南充段)流域8个污水处理厂的进水、尾水以及附近水样品,测定了常规水质指标,并利用Illumina NovaSeq 6000平台对水体微生物群落进行测序分析。结果显示:pH为附近水>进水>尾水,TN、NH_(4)^(+)-N、TP为进水>尾水>附近水。尾水中的微生物多样性高于进水和附近水;所有水体中,微生物均以变形菌门(Proteobacteria)为主,但不同来源水体群落组成显著不同;相较于附近水,尾水中微生物具有更多的特有种。此外,相关性分析结果表明,附近水的微生物群落与水体理化因子相关性最大,且仅附近水的微生物多样性指数与理化因子相关。综合来看,嘉陵江南充段城镇污水厂尾水的微生物可作为生物源影响江水微生物,且尾水亦能通过提升水体营养间接影响江水微生物群落组成。城镇污水厂应该进一步改进净水工艺,降低尾水水体营养,同时提升杀菌强度,从两方面来避免沿岸城镇污水处理厂尾水对嘉陵江微生物群落的潜在影响。

Effects of dietary supplementation of bacteriophage cocktail on health status of weanling pigs in a non-sanitary environment

Background The study evaluated the effects of bacteriophage cocktail(BP)and ZnO administered during weaning time for piglets exposed to a non-sanitary environment.The bacteriophages were designed to eliminate Escherichia coli(K88,K99 and F41),Salmonella(typhimurium and enteritidis),and Clostridium perfreingens(types A and C).Forty 21-day-old crossbreed piglets were assigned to four treatments,including the PC(sanitary environment),NC(nonsanitary environment),BP(NC plus 108 pfu/kg BP),and ZO(NC plus 2,500 mg/kg ZnO).Piglets in the NC,BP and ZO were kept in a non-sanitary environment for 14 d,which was contaminated with the feces of infected pigs.Results Pigs in the BP and ZO treatments had a higher final body weight compared with the NC.The NC treatment showed the highest concentration of inflammatory cytokines including interleukin(IL)-1β,IL-6 and tumor necrosis factor-αin the plasma.The administration of BP and ZO showed lower myeloperoxidase concentrations compared with the NC.The NC treatment showed a lower concentration of superoxide dismutase in serum compared with the PC.Among the treatments in non-sanitary environment,the NC treatment showed a higher concentration of malondialdehyde compared with the ZO.The PC treatment showed a lower concentration of butyric acid in the feces compared with the BP treatment.Among non-sanitary treatments,the villus height in the duodenum was greater in the BP and ZO compared with the NC.The lower abundance of Proteobacteria phylum was observed in the BP and PC treatments compared with the NC.The highest relative abundance of Eubacterium was recorded in the BP treatment.The abundance of Megasphaera and Schwartzia was higher in the NC pigs compared with the BP piglets.The abundance of Desulfovibrio was lower in the supplemented treatments(BP and ZO)compared with non-supplemented(NC and PC).The abundance of Cellulosilyticum genera was higher in the BP and ZO treatments rather than in the NC.The piglets in the NC treatment had the highest abundance of Escherichia-Shigella,followed by the PC and ZO treatments.Conclusion In conclusion,these results suggest that the supplementation of bacteriophage cocktail could effectively control Proteobacteria phylum,Clostridium spp.and coliforms population and mitigated the adverse influences of weaning stress in piglets.

16SrDNA克隆文库方法分析MDAT-IAT同步脱氮除磷系统细菌多样性研究

采用分子生物学手段16SrDNA克隆文库方法对高效同步脱氮除磷系统MDATIAT(modifieddemandaerationtankintermitaerationtank)的IAT池中的细菌进行了多样性研究.从16SrDNA克隆文库中随机挑选59个克隆子进行序列测定(约500bp),对测序结果进行了BLAST比对.结果表明,MDATIAT系统中IAT池的细菌群落具有高度多样性,有55个克隆子分属6个不同的细菌类群,3个克隆子属于未知类群,优势细菌类群为Proteobacteria类群(变形菌类群),占55.17%;细菌类群优势顺序为γProteobacteria类群(34.48%)、Bacteroidetes类群(似杆菌类群,20.69%)、βProteobacteria类群(12.07%)、CandidatedivisionTM7类群(12.07%)、αProteobacteria类群(5.17%)、δProteobacteria类群(3.45%)、Firmicutes类群(厚壁菌类群,3.45%)、CandidatedivisionOP11类群(1.72%)、Planctomycetes类群(浮霉状菌类群,1.72%).用clustalx软件从测序的58个克隆子中选出32个OTU进行系统发育分析,同样表明IAT池的细菌多样性较强.

秸秆还田后土壤微生物群落结构变化的初步研究

通过实验室条件下的小麦秸秆粉和油菜秸秆粉的还土试验,结合常规的分析手段和基于DNA的分子技术(变性梯度凝胶电泳(DGGE)及测序技术),初步研究了秸秆粉还田后土壤物理化学特性及生物学特性的变化。结果表明,培养60d后,秸秆还田土壤的肥力明显提高,其纤维素酶活性明显增强。DGGE图谱表明,对照土壤(S)以及处理土壤(SW和SR),在培养过程中,β-Proteobacteria类细菌组成都在发生变化。对其中一个样品的DGGE条带进行了割胶测序,测序结果表明,大部分条带代表的微生物是未培养的或不可培养的。采用传统分离培养技术,从秸秆还田土壤中分离了2株纤维素降解菌。以上结果说明,秸秆还田具有良好的土壤综合效应。

环境因素对甲基叔丁基醚生物降解的影响研究

采用β-Proteobacteria PM1完整细胞降解甲基叔丁基醚(MTBE),研究了金属离子、pH值、细胞浓度等因素对MTBE降解速率的影响.结果表明,K+和Ba2+对MTBE降解有明显的促进作用,较适宜的pH值为7.0;细胞在BaCl2溶液中基本上处于静息状态;细胞浓度越高,降解MTBE的速率就越快;降解过程需要有氧气参与;PM1完整细胞降解叔丁醇(TBA)的速率比降解MTBE更快,MTBE的存在对TBA的降解有抑制作用,但TBA的存在不影响MTBE的降解;静息细胞法降解MTBE的过程中没有检测到TBA等中间产物;PM1静息细胞降解MTBE的米氏常数Km为0.73mmol·L-1,最大反应速率Vmax为0.14mmol·L-1·h-1.

基于PCR-DGGE技术分析生物絮团的细菌群落结构

在草鱼养殖过程中添加碳源(葡萄糖)维持水体C∶N为20∶1以培养生物絮团,通过对生物絮团细菌群落构成进行种类鉴定来评价生物絮团中功能微生物的组成。采用PCR-DGGE(变性梯度凝胶电泳)技术分析生物絮团形成第5、10和15天的细菌群落结构。DGGE指纹图谱结果分析表明:第5天和第10天的相似性最高,达67.4%;第5天和第15天相似性系数最低仅为40.5%。第10天时微生物多样性最高,第15天时多样性最低。对DGGE指纹图谱特征条带进行回收、克隆测序,结果表明,生物絮团培养过程主要微生物类群隶属于以下6个纲:α-变形菌纲(Alphaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)、β-变形菌纲(Betaproteobacteria)、放线菌纲(Actinobacteria)、芽孢杆菌纲(Bacilli)、拟杆菌纲(Bacteroidetes),其中α-、β-及γ-变形菌占据主要位置。α-proteobacteria为3个阶段的共有优势菌,第5天时特异菌包括食酸菌属(Acidovorax)、气单胞菌属(Aeromonas)、土壤杆菌属(Agrobacterium),第10天和15天分别为芽孢杆菌属(Bacillus)与红球菌属(Rhodococcus)。研究首次发现,生物絮团应用于淡水养殖系统时细菌的组成和多样性都极其丰富,通过结合分析这些微生物的功能特点,为生物絮团技术在实际养殖生产中的进一步应用奠定基础。

黄海西北部沉积物中细菌群落16S rDNA多样性解析

为揭示黄海西北部海域不同站位沉积物中细菌群落的多样性,采用16SrDNA文库技术,对黄海西北部辽东半岛和山东半岛近岸5个站位的沉积物中不同季节的细菌群落特征进行解析和评价.结果表明,沉积物中微生物种类丰富,主要分布于5～10个已知的门,另外还存在大量未被认知的序列;各站位沉积物中优势菌均为变形细菌门,占文库序列的46%～60%,其中γ-和δ-变形细菌纲为门中优势类群,但在各个站位中存在明显系统发育学分歧.微生物种群在不同地理区域和季节都存在明显差异,不同功能类群与其特殊生境密切相关.

不同干扰方式对喀斯特生态系统土壤细菌优势类群—变形菌群落的影响

以喀斯特原生林为对照,运用16S rRNA基因的PCR-RFLP和测序技术对该区不同人为干扰方式下土壤细菌的群落结构进行了分析。结果显示,4个样地中变形菌占总克隆子数的41.3%,是研究区土壤中的优势细菌类群。与原生林地相比较,受人为干扰的生态系统土壤中变形菌明显减少,自然恢复地、农耕地和放牧+冬季火烧草地减少了30.2%～47.4%。自然恢复地、放牧+冬季火烧草地与原生林地土壤中变形菌的4个亚群丰度分布关系一致,均为α->δ->β->γ-变形菌,而农耕地则为δ->α->β->γ-变形菌,说明自然恢复和放牧+冬季火烧草地对喀斯特土壤变形菌的恢复作用有限,而对变形菌4个亚群之间的分布关系有明显的正效应,尤其是自然恢复地中α-变形菌得到了很好的恢复,较农耕地增加了130%。四个样地中,占总克隆子数16.5%的克隆子被归类为根瘤菌目,且以原生林地最多,是3个干扰样地的1.6～3.7倍。基于以上研究结果,未来可考虑种植本土固氮植物结合接种相应的固氮微生物作为恢复喀斯特退化生态系统的措施之一。

云南4处酸性热泉中的变形菌门细菌多样性

选择云南腾冲和龙陵地区的4处酸性热泉（pH：2．3—6．0，温度：47～96℃），通过Barcoded pyrosequencing技术及统计学分析，详细阐述了这些样点中变形菌门的物种和遗传多样性。本研究共获得了2489条变形菌门16SrRNA基因序列，可划分为234个可操作分类单元（OTUs）。分类结果显示，热泉内涵盖了大量已知的目、科和属。而41％的OTUs可能代表了变形菌门的未知物种。系统发育分析显示，样点中存在着若干全新遗传类群，这些样点和美国黄石地区的热泉各自呈现不同的变形菌门遗传多样性。此外，本次调查的热泉彼此间存在着变形菌门群落结构的显著差异。

秋、冬季刺参养殖池塘菌群的多样性分析

以黄海和渤海代表性刺参养殖池塘秋、冬季海水和沉积物基因组DNA为模板,以细菌16S r DNA通用引物进行PCR扩增,构建16S r DNA文库并进行测序分析,研究了秋、冬季刺参养殖池塘菌群的多样性。结果表明:海水和沉积物中主要包括11个门类的细菌,即变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、疣微菌门(Verrucomicrobia)、厚壁菌门(Firmicutes)、梭杆菌门(Fusobacteria)、蓝细菌门(Cyanobacteria)、浮霉菌门(Planctomycetes)、脱铁杆菌门(Deferribacteres)、绿弯菌门(Chloroflexi);黄海和渤海刺参养殖池塘海水和沉积物中优势类群均为变形菌(变形菌比例>48%);用Shannon指数及Simpson优势度指数分析细菌的多样性,黄、渤海刺参养殖池塘中,冬季沉积物细菌Simpson优势度指数均最低,分别为0.014 89和0.016 50,Shannon指数均最高,分别为6.312和5.695。研究表明,黄海和渤海刺参养殖池塘中,冬季沉积物细菌多样性均最高。

除磷工艺中含氧条件对聚磷菌种群结构影响研究

利用分子生物学技术直接从活性污泥样品中提取DNA,采用套式PCR技术对特征基因片断进行扩增,结合DGGE(变性浓度梯度凝胶电泳)分析活性污泥中微生物种群结构.研究了除磷工艺在正常运行情况下的微生态系统种群结构,并分析了微生物群落结构及行为特征.测定了活性污泥中变形杆菌(Proteobacteria)和产酸菌(Acidobacterium)部分菌种的16S rDNA V3区片段序列,通过NCBI(美国国立生物技术信息中心)基因库比对,初步确定了部分细菌的属.在厌氧/好氧和缺氧/好氧工艺中各类优势菌群变化规律研究表明,在除磷效果稳定的情况下,系统中除磷微生物种群结构大致能保持不变,少数数量或种类发生变化的种群与系统中含氧量变化有关,但是处于动态变化中的菌群结构总体能够适应工艺运行环境.

FISH技术解析不同氨氮浓度MBR中的微生物群落结构

为了探究膜生物反应器(MBR)中微生物群落结构与硝化作用的内在关系,采用荧光原位杂交技术(FISH)对不同氨氮浓度的MBR系统中微生物种群进行检测,考察变形菌(Proteobac-teria)、放线菌(Actinobacteria)、黄杆菌(Cytophaga-flexibacter-bacteroides)、绿弯菌(Chloroflexi)、硝化细菌(Nitrobacter sp.)和氨氧化细菌(Ammonia-oxidizing bacteria)占总细菌的相对含量.结果表明:进水氨氮浓度分别为35,3.47和100 mg/L的MBR系统中,变形菌分别占总细菌的51%,44%和39%,氨氧化细菌与硝化细菌之和分别占总细菌的28%,4%和43%.在氨氮浓度较低的MBR中,检测到大量丝状β-变形菌,同时放线菌和绿弯菌也有所增加.高氨氮负荷有利于氨氧化细菌和硝化细菌的生长富集,并使其在系统中成为优势种群.相关性分析表明,氨氧化细菌、硝化细菌与氨氮去除呈显著正相关.

成年与未成年圈养林麝粪便菌群多样性的比较

本研究旨在解析林麝未成年组(n=10)和成年组(n=10)之间的菌群差异。收集林麝新鲜粪便,提取总DNA,利用带标签的通用引物扩增16S rRNA V3-V4区,使用Illumina Miseq 300PE测序平台对扩增产物进行Miseq双端测序,通过计算ACE和Shannon等多样性指数,以及微生物组成成分和距离聚类分析,揭示组成结构与差异。α多样性分析表明成年组微生物的多样性丰富度略高于未成年组,但不存在显著差异性(P>0.05)。未成年组和成年组均是厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和变形菌门(Proteobacteria)所占的比例较高。成年组Proteobacteria比例明显高,而Firmicutes比例较低。LEfSe分析表明有12个显著差异的细菌分类。两组在Firmicutes门存在显著差异的细菌较少,而在Proteobacteria上存在显著差异的细菌较多。本研究证明在不同年龄阶段,微生物的丰富度和多样性不存在显著差异,细菌的组成成分也相同。但是在组成比例上存在差异性,间接反映了不同年龄阶段营养吸收的不同需求。

青岛汇泉湾海洋趋磁细菌多样性研究

在青岛汇泉湾沿岸沉积物中发现了大量海洋趋磁细菌,最大丰度可达10~5个/cm^3。透射电镜观察发现该菌菌体形态多样,有球形或卵球形、长短杆状、弧状和螺旋状,其中球形或卵球形趋磁细菌占绝对优势。电镜观察还发现该菌磁小体的排列方式多样化,大多数呈链状排列,有单链、双链及多链,还有的呈环状或者成簇排列。磁小体的形态也多种多样,有正方体、棱柱体、立方八面体、子弹头状、片状和齿状。用 RFLP 方法分析了70个克隆测序,得到10条不同序列。经16S rRNA 系统发育分析,发现9个属于α-变形菌亚纲,1个属于γ-变形菌亚纲,共有8个不同的属,优势种属于未培养的海洋趋磁球菌。所有菌株与最接近的海洋趋磁球菌的相似性并不高(76.4%～89.4%),表明该海区的趋磁细菌为新发现的微生物资源。