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From the Mountain to the Valley - Drivers of Groundwater Prokaryotic Communities along an Alpine River Corridor 从山到谷——高山河流走廊沿线地下水原核生物群落的驱动因素
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108227
Clemens Karwautz, Alice Retter, Johannes Haas, Steffen Birk, Christian Griebler
Rivers extend into the underlying groundwater which represents the unseen freshwater majority. Understanding microbial community composition and dynamics of shallow groundwater and lotic ecosystems is thus crucial, due to their potential impact on ecosystem processes and functioning. A 300 km section of the Mur river valley, from 2000 m.a.s.l. in the Austrian alps to the flats (200 m.a.s.l.) at the Slovenian border, was followed, analyzing river water from 14 stations and groundwater from 45 wells in early summer and late autumn. The active (RNA derived) and total prokaryotic communities were characterized using high-throughput gene amplicon sequencing. Key physico-chemical parameters and stress indicators were recorded. The groundwater microbiome is analyzed regarding its composition, change with land use, and difference to the river. Community composition and species turnover differed significantly. At high altitudes, dispersal limitation was the main driver of groundwater community assembly, whereas in the lowland, homogeneous selection explained the larger share. Land use was a key determinant of the groundwater microbiome composition. The alpine region was more diverse in prokaryotic taxa, with some early diverging archaeal lineages being highly abundant. This dataset shows a longitudinal change in prokaryotic communities that is dependent on regional differences affected by geomorphology and land use.
河流延伸到地下地下水,这代表了看不见的大部分淡水。因此,了解浅层地下水和河流生态系统的微生物群落组成和动态至关重要,因为它们对生态系统过程和功能有潜在影响。研究人员跟踪了从奥地利阿尔卑斯山脉2000年到斯洛文尼亚边境平原(200年)的穆尔河谷300公里的一段,分析了初夏和深秋期间14个站点的河水和45口井的地下水。利用高通量基因扩增子测序对活性(RNA衍生)和总原核生物群落进行了表征。记录了关键的理化参数和应力指标。分析了地下水微生物群的组成、随土地利用的变化以及与河流的差异。群落组成和物种更替差异显著。在高海拔地区,分散限制是地下水群落聚集的主要驱动力,而在低海拔地区,均匀选择解释了更大的份额。土地利用是地下水微生物组成的关键决定因素。高寒地区的原核生物类群更为多样化,一些早期分化的古细菌谱系非常丰富。该数据集显示了原核生物群落的纵向变化,这种变化依赖于受地貌和土地利用影响的区域差异。
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引用次数: 0
A global atlas of subsurface microbiomes reveals phylogenetic novelty, large scale biodiversity gradients, and a marine-terrestrial divide 地下微生物组的全球地图集揭示了系统发育的新颖性,大规模的生物多样性梯度和海洋-陆地鸿沟
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108531
S Emil Ruff, Isabella Hrabe de Angelis, Megan Mullis, Jerome Payet, Brandi Kiel Reese, Karen Lloyd, Andrew Steen, Hilary Morrison, Mitchell Sogin, Joshua Ladau, Frederick Colwell
Marine and terrestrial subsurface sediments, rocks, and water may represent the largest habitat for microbial life on Earth. Despite the global importance of subsurface ecosystems for biogeochemical cycling and microbial diversity, essential questions remain unanswered. These concern the abundance of novel microbial clades in the subsurface, the difference between marine and terrestrial microbiomes, as well as between subsurface and surface microbiomes, and the adaptation of specific clades to environmental conditions of the subsurface. Here, we analyzed 523 archaeal and 1,211 bacterial 16S rRNA gene amplicon sequence datasets and 146 shotgun metagenomes from surface, interface, and subsurface ecosystems worldwide, including lakes, saltmarshes, hot springs, caves, mines, methane seeps, hydrothermal vents, and seafloor sediments. We found that archaeal and bacterial alpha diversity (per sample richness and evenness) and beta diversity (community differentiation) varied continuously between surface and subsurface biomes, but differed abruptly between marine and terrestrial subsurface ecosystems. Bacterial alpha diversity tended to be lower in subsurface than surface ecosystems. However, subsurface archaeal alpha diversity often exceeded that of surface ecosystems suggesting that the subsurface holds a considerable and largely underestimated fraction of Earth’s archaeal diversity. Overall, microbial communities of marine subsurface ecosystems exhibited greater alpha diversity, while gamma diversity (total richness and evenness) was higher in the terrestrial subsurface, potentially due to greater habitat diversity. We identify diagnostic clades, especially for the archaea, that are widespread in marine (e.g., Lokiarchaeia, Bathyarchaeia) and terrestrial (e.g., Hadarchaeia, Methanococci) subsurface ecosystems. The substantial community overlap along depth transects and between surface, interface and subsurface realms suggests a global gradient between the surface and subsurface rather than a discrete and defined subsurface biosphere. Finally, none of the included subsurface ecosystems seem to be exhaustively sampled, leaving much biodiversity and metabolic capability yet to be discovered.
海洋和陆地的地下沉积物、岩石和水可能是地球上微生物生命的最大栖息地。尽管地下生态系统对生物地球化学循环和微生物多样性具有全球重要性,但基本问题仍未得到解答。这些问题涉及地下新微生物分支的丰度,海洋和陆地微生物组之间的差异,以及地下和地表微生物组之间的差异,以及特定分支对地下环境条件的适应。在这里,我们分析了523个古细菌和1211个细菌16S rRNA基因扩增子序列数据集和146个鸟枪宏基因组,这些数据集来自世界各地的地表、界面和地下生态系统,包括湖泊、盐沼、温泉、洞穴、矿山、甲烷渗漏、热液喷口和海底沉积物。研究发现,古细菌和细菌的α多样性(每样丰富度和均匀度)和β多样性(群落分化)在地表和地下生态系统之间持续变化,而在海洋和陆地地下生态系统之间差异明显。细菌α多样性在地下生态系统中低于地表生态系统。然而,地下古细菌的α多样性经常超过地表生态系统,这表明地下拥有相当大的古细菌多样性,而且在很大程度上被低估了。总体而言,海洋地下生态系统的微生物群落表现出更大的α多样性,而陆地地下生态系统的伽马多样性(总丰富度和均匀度)更高,这可能是由于更大的栖息地多样性。我们确定了广泛存在于海洋(如Lokiarchaeia, Bathyarchaeia)和陆地(如Hadarchaeia, Methanococci)地下生态系统中的诊断分支,特别是古细菌。沿着深度样带以及地表、界面和地下领域之间的大量群落重叠表明地表和地下之间存在全球梯度,而不是一个离散的、明确的地下生物圈。最后,所有的地下生态系统似乎都没有得到详尽的采样,留下许多生物多样性和代谢能力有待发现。
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引用次数: 0
Non-Deterministic Factors Affect Competition Between Thermophilic Autotrophs from Deep-Sea Hydrothermal Vents 不确定性因素影响深海热液喷口嗜热自养生物之间的竞争
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108248
Briana Kubik, James Holden
Hydrothermal vents provide windows into the rocky subseafloor on Earth and serve as terrestrial analog sites for extraterrestrial environments. By studying patterns of community assembly in hydrothermal vents and using geochemical models, we can better understand how the deep-sea biosphere contributes to local and global biogeochemical cycling and gather valuable information about how similar communities may arise on Earth and beyond Earth. One prevailing thought is that vent microbial community assembly is driven by deterministic factors such as the thermodynamic favorability of redox reactions. We hypothesized that subsurface microbial communities may also be significantly influenced by other factors, such as differential cell yields, varying optimal growth temperatures, and stochasticity. At Axial Seamount in the Pacific Ocean, H 2 -consuming methanogens of the genera Methanocaldococcus (T opt 82°C) and Methanothermococcus (T opt 65°C) and H 2 -consuming sulfur reducers of the genus Desulfurobacterium (T opt 72°C) are the most abundant autotrophs that grow optimally at or above 65°C (Fortunato et al. 2017). At one low-temperature hydrothermal vent site, Marker 113, methanogens are the predominant thermophilic autotrophs while at another site, Marker 33, thermophilic autotrophic sulfur reducers predominate. There is no apparent geochemical or thermodynamic explanation for the differences in community composition. In this study, we performed a series of co-culture competition experiments using Methanocaldococcus jannaschii , Methanothermococcus thermolithotrophicus , and Desulfurobacterium thermolithotrophum HR11 as representative methanogens and sulfur reducers common to hydrothermal vents to explain the variations in community composition between thermophilic autotrophs. M. jannaschii increases its cell yield (cells produced per mole of CH 4 produced) when grown on very low H 2 concentrations as part of a growth rate-growth yield tradeoff (Topçuoğlu et al. 2019). This increase in cell yield could provide methanogens with a competitive growth advantage over H 2 -consuming sulfur reducers, who otherwise catalyze a more thermodynamically favorable growth reaction. Competition co-culture experiments were conducted between M. jannaschii and D. thermolithotrophum at 72°C and between M. thermolithotrophicus and D. thermolithotrophum at 65°C, both at 1:1 ratios and initial aqueous H 2 concentrations of 1.2 mM (high H 2 ) and 85 μM (low H 2 ) to determine the effects of temperature and H 2 availability on autotroph competition. For both methanogens, the growth rate, maximum cell concentration, and total CH 4 produced decreased when they were grown in co-culture, at low H 2 , or both relative to monocultures grown with high H 2 . The methanogen cell yields generally increased in co-culture and at low H 2 . At both experimental temperatures, the growth rate of D. thermolithotrophum remained unchanged in co-culture and at low H 2 relative to monocultures
热液喷口提供了进入地球岩石海底的窗口,并充当了地球上对地外环境的模拟站点。通过研究深海热液喷口的生物群落组合模式并使用地球化学模型,我们可以更好地了解深海生物圈如何促进本地和全球生物地球化学循环,并收集有关地球上和地球以外类似群落如何出现的有价值的信息。一种普遍的观点认为,喷口微生物群落的聚集是由确定性因素驱动的,如氧化还原反应的热力学有利性。我们假设地下微生物群落也可能受到其他因素的显著影响,如不同的细胞产量、不同的最佳生长温度和随机性。在太平洋轴向海山,耗氢产甲烷菌Methanocaldococcus (T opt 82°C)和耗氢产热球菌(T opt 65°C)和耗氢产硫菌Desulfurobacterium属(T opt 72°C)是最丰富的自养菌,在65°C或更高温度下生长最佳(Fortunato et al. 2017)。在一个低温热液喷口位置,标记113,产甲烷菌是主要的嗜热自养生物,而在另一个位置,标记33,嗜热自养硫还原剂占主导地位。群落组成的差异没有明显的地球化学或热力学解释。本研究以jannaschii甲烷钙球菌(Methanocaldococcus jannasii)、热养甲烷球菌(Methanothermococcus thermolithotrophum HR11)和热养Desulfurobacterium thermolithotrophum HR11为代表热液喷口常见的产甲烷菌和硫还原剂,进行了一系列共培养竞争实验,以解释嗜热自养菌之间群落组成的差异。作为生长速率-生长产量权衡的一部分,在极低的h2浓度下生长时,M. jannaschii增加了细胞产量(每摩尔产生的甲烷产生的细胞数)(Topçuoğlu et al. 2019)。这种细胞产量的增加可以为产甲烷菌提供比消耗h2的硫还原剂更具竞争力的生长优势,否则后者催化的生长反应在热力学上更有利。在72°C条件下,jannaschii与d.m othotrophium在72°C条件下,以及65°C条件下,在1:1的比例和初始水浓度分别为1.2 mM(高H 2)和85 μM(低H 2)条件下,进行了竞争共培养实验,以确定温度和H 2有效度对自养菌竞争的影响。对于这两种产甲烷菌,在共培养、低H条件下或在高H条件下单独培养时,其生长速率、最大细胞浓度和总甲烷产量均下降。在共培养和低H条件下,产甲烷菌的产量普遍增加。在两种实验温度下,与单培养相比,共培养和低h2o2条件下,热石营养菌的生长速度保持不变,但在两种h2o2条件下,共培养的最大细胞浓度均低于单培养。然而,在低H 2条件下,无论是单培养还是共培养,都没有检测到硫还原剂产生的H 2 S,这表明生长产量发生了显著变化。在温度和h2浓度下,硫还原剂的细胞浓度均高于产甲烷菌。随机性或喷口流体化学可能导致产甲烷菌早期在喷口定殖,随后由于产甲烷菌的数量优势而将自养硫还原剂排除在生态位之外。因此,在高H条件下,在不同的初始甲烷菌与硫还原剂比例下,进行竞争性共培养实验。在72℃时,单培养和共培养的D. thermolithotrophum达到了相同的最大细胞浓度,产生了相同数量的h2s,即使产甲烷菌最初的数量是硫还原菌的1万倍。在所有共培养中,与单培养相比,jannaschi达到了较低的最大细胞浓度,产生的ch4较少。在65℃时,D. thermolithotrophum在单培养和共培养中达到了相同的最大细胞浓度,产生了相同数量的h2s,其中产甲烷菌的数量最初超过了硫还原菌的100倍。然而,当产甲烷菌的数量最初超过还原剂1000倍时,热石营养菌在单培养条件下生长良好,其最大细胞浓度和产生的h2s量显著低于单培养和其他共培养条件。综上所述,当h2受限时,产甲烷菌和硫还原剂的氧化还原反应分别从生成ch4和h2s转向生成生物质。这应该在热力学预测模型中加以考虑。 此外,低于最佳硫还原剂的生长温度和相对于硫还原剂的高初始产甲烷菌浓度的组合可以通过生态位排斥导致产甲烷菌在排气环境中长期优于自养型硫还原剂。
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引用次数: 0
Differences Between Heterotrophic and Nitrate-dependent Iron-oxidizing Microbial Communities in Bioreactor Sediment Treating Mine Wastewater 异养型和硝酸盐依赖型生物反应器沉积物中铁氧化微生物群落的差异
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108177
Hannah Koepnick, Brent Peyton, Ellen Lauchnor
Nitrate-dependent iron oxidation (NDFO) is a novel mechanism for microbial bioremediation of metal and metalloid contaminants. During NDFO, microbes catalyze a redox reaction wherein nitrate is reduced to nitrite and nitrogen gas while Fe(II) is oxidized to solid Fe(III) hydroxide minerals. Metalloid contaminants such as selenium and arsenic have a propensity for adsorption to iron minerals produced during NDFO; some contaminants may also be concurrently bioreduced. A number of bacterial isolates have been shown to be capable of NDFO (e.g., Kappler et al. (2005), Kiskira et al. (2017)), but little work has been done to date characterizing mixed microbial communities performing NDFO. Some autotrophic communities have been characterized, with high relative abundances for strains of Gallionellaceae in both a freshwater sediment enrichment culture and an activated sludge culture (Blöthe and Roden 2009, Tian et al. 2020). In mixotrophic activated sludge cultures, the concentration of Fe(II) amendment was found to significantly impact microbial community composition; these cultures were fed with methanol in addition to Fe(II), and the dominant community members were Methyloversatilis and other methylotrophic strains (Liu et al. 2018). The work presented here examines microbial communities performing NDFO in the context of remediation, and in particular how differences between NDFO and heterotrophic communities may influence remediation effectiveness. This research characterizes and compares microbial communities performing NDFO versus heterotrophic denitrification during removal of selenium and nickel from mining wastewater. Sediment and influent water from a subsurface bioreactor treating mining wastewater were used to construct batch bioreactors, which were amended with selenium and nickel as well as either Fe(II) or methanol to investigate contaminant removal and microbial community composition in NDFO versus heterotrophic microbial communities. Both Fe(II) and methanol reactors removed total aqueous selenium to below the quantification limit, but Fe(II) reactors removed it more rapidly, likely due to adsorption of selenite. For nickel, removal to below the detection limit was achieved with methanol amendment, while Fe(II) amendment resulted in 42-95% removal. This was likely due to precipitation of nickel sulfide during sulfate reduction in methanol-amended reactors. DNA from the batch bioreactors will be sequenced and the results analyzed for differences among communities. Permutational multivariate analysis of variance and non-metric multidimensional scaling will be used to determine significant correlations of community composition with experimental variables, selenium and nickel removal, and NDFO (Roberts 2023, Kruskal 1964). Indicator species analyses (De Cáceres et al. 2010) will be applied to identify taxa found significantly more often (i.e., at a higher relative abundance) in one group of microbial communities than in any other group. The
硝酸盐依赖铁氧化(NDFO)是微生物修复金属和类金属污染物的一种新机制。在NDFO过程中,微生物催化氧化还原反应,其中硝酸盐被还原为亚硝酸盐和氮气,而铁(II)被氧化成固体铁(III)氢氧化物矿物。类金属污染物,如硒和砷,有吸附倾向的铁矿物产生的NDFO;一些污染物也可以同时被生物还原。许多细菌分离株已被证明能够进行NDFO(例如,Kappler等人(2005),Kiskira等人(2017)),但迄今为止,对执行NDFO的混合微生物群落进行表征的工作很少。一些自养群落的特征是,在淡水沉积物富集培养和活性污泥培养中,Gallionellaceae菌株的相对丰度都很高(Blöthe and Roden 2009, Tian et al. 2020)。在混合营养活性污泥培养中,发现铁(II)修正浓度显著影响微生物群落组成;在这些培养物中添加甲醇和Fe(II),优势群落成员是methylomultilis和其他甲基营养菌株(Liu et al. 2018)。本文介绍的工作考察了在修复背景下执行NDFO的微生物群落,特别是NDFO和异养群落之间的差异如何影响修复效果。本研究表征并比较了在去除采矿废水中的硒和镍过程中进行NDFO和异养反硝化的微生物群落。利用处理采矿废水的地下生物反应器的沉积物和进水构建间歇式生物反应器,并对其进行硒和镍以及铁(II)或甲醇的改性,以研究NDFO与异养微生物群落的污染物去除和微生物群落组成。Fe(II)反应器和甲醇反应器对水中总硒的去除率均低于定量限制,但Fe(II)反应器的去除率更快,可能是由于亚硒酸盐的吸附。甲醇对镍的去除率低于检出限,而Fe(II)的去除率为42-95%。这可能是由于在甲醇改性反应器中硫酸盐还原过程中硫化镍的沉淀。将对间歇式生物反应器的DNA进行测序,并对结果进行分析,以确定不同群落之间的差异。将使用置换多元方差分析和非度量多维尺度来确定群落组成与实验变量、硒和镍去除以及NDFO之间的显著相关性(Roberts 2023, Kruskal 1964)。指示物种分析(De Cáceres et al. 2010)将用于识别在一组微生物群落中比在任何其他组中更频繁(即相对丰度更高)发现的分类群。指示种分析可以揭示在NDFO条件下占优势的反硝化菌群与在异养反硝化过程中占优势的反硝化菌群之间是否存在差异。这些微生物群落分析的结果,结合地球化学分析,将提高我们对在修复环境中执行NDFO的微生物群落的理解。
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引用次数: 0
Phytoplankton Ecology in an Oilsands End Pit Lake 油砂尾坑湖浮游植物生态学研究
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108385
Chantel C. Furgason, Angela Smirnova, Joel Dacks, Peter Dunfield
Alberta oilsands mining and extraction have produced over 1 trillion litres of tailings wastewater (AER 2021) containing several compounds of concern (Cossey et al. 2021). End-pit lakes are a low-cost, long-term proposed strategy of tailings reclamation that sequester tailings in a mined-out pit under a freshwater cap. Through dilution and biogeochemical processes, the water cap should over time develop into a functional ecosystem integrable with the local watershed (Cossey et al. 2021, Saborimanesh 2021). Established in 2012, Base Mine Lake is currently the only full-scale pilot end pit lake developed by the Alberta oilsands industry and requires further investigation to validate end pit lakes as a tailings reclamation technology (Cossey et al. 2021). The first stage of reclamation requires the development of a phytoplankton community, which serves as the base of the aquatic food web (CEMA 2012). The primary objective of this study was to characterize the phytoplanktxon community over time in BML from 2016 to 2021 to determine how community composition and abundances shift over seasons and years. Characterization used Illumina gene sequencing targeting 16S rRNA, 18S rRNA, and 23S rRNA gene amplicons, giving relative abundance data over time for phytoplankton. Cell count data was used to verify gene sequencing results. The phytoplankton community composition and diversity in Base Mine Lake was compared to those of a freshwater reservoir and a tailings pond. Analysis of gene sequencing data revealed that major genera of phytoplankton included Cryptomonas ( Cryptophyceae ), Choricystis ( Trebouxiophyceae ), Euglena ( Euglenales ), and Synechococcus ( Synechococcales ), all of which appear to exhibit seasonal blooms during 2016-2021 (Fig. 1). Sequencing analysis also indicated that Base Mine Lake and its freshwater input source Beaver Creek Reservoir shared many of the same genera but different strains/species of those genera. This suggested that the distinct conditions in each aquatic site may have selected for distinct strains. Diversity analyses of gene sequencing data revealed that phytoplankton diversity in Base Mine Lake was intermediate between that of its freshwater input reservoir and a tailings pond. There currently exists no low-cost, large-scale treatment method that fully reclaims tailings water (Cossey et al. 2021, Saborimanesh 2021). End-pit lakes retain tailings until recalcitrant compounds are degraded to near-environmental levels (Saborimanesh 2021, CEMA 2012), but further research is required before end-pit lakes can be approved as a viable reclamation technology (Cossey et al. 2021). Although research is available now on native microbial communities in tailings waters, knowledge on their contributions to an aquatic microbial food web is limited (Saborimanesh 2021). This proposed research is the first of its kind to examine the contribution of phytoplankton to end pit lake food web ecology. This will advance knowledge of end-pit l
艾伯塔省油砂开采和提取产生了超过1万亿升的尾矿废水(AER 2021),其中含有几种令人担忧的化合物(Cossey et al. 2021)。尾坑湖是一种低成本、长期的尾矿回收策略,它将尾矿隔离在淡水帽下的采出坑中。通过稀释和生物地球化学过程,随着时间的推移,水帽将发展成为与当地流域可整合的功能性生态系统(Cossey et al. 2021, Saborimanesh 2021)。Base Mine Lake成立于2012年,是目前阿尔伯塔省油砂行业开发的唯一一个全面的试验性尾坑湖,需要进一步调查以验证尾坑湖作为尾矿回收技术的有效性(Cossey et al. 2021)。复垦的第一阶段需要发展浮游植物群落,这是水生食物网的基础(CEMA 2012)。本研究的主要目的是表征2016年至2021年BML浮游植物群落随时间的变化,以确定群落组成和丰度如何随季节和年份变化。利用Illumina基因测序对16S rRNA、18S rRNA和23S rRNA基因扩增子进行鉴定,获得浮游植物随时间变化的相对丰度数据。细胞计数数据用于验证基因测序结果。将基地矿湖的浮游植物群落组成和多样性与淡水水库和尾矿库进行了比较。基因测序数据分析显示,浮游植物的主要属包括隐单胞菌(Cryptophyceae)、毛囊藻(Trebouxiophyceae)、藻芽藻(Euglenales)和聚球菌(Synechococcales),它们在2016-2021年期间都出现了季节性华(图1)。测序分析还表明,Base Mine Lake及其淡水输入源Beaver Creek Reservoir具有许多相同的属,但这些属的菌株/种不同。这表明,每个水生地点的不同条件可能选择了不同的菌株。基因测序数据的多样性分析表明,基矿湖浮游植物多样性介于其淡水输入库和尾矿库之间。目前还没有低成本、大规模的完全回收尾矿水的处理方法(Cossey et al. 2021, Saborimanesh 2021)。尾坑湖保留尾矿,直到顽固性化合物降解到接近环境的水平(Saborimanesh 2021, CEMA 2012),但在尾坑湖被批准为可行的回收技术之前,还需要进一步的研究(Cossey et al. 2021)。虽然目前对尾矿水中的原生微生物群落进行了研究,但对它们对水生微生物食物网的贡献的了解有限(Saborimanesh 2021)。本研究首次探讨了浮游植物对尾坑湖食物网生态的贡献。这将促进对尾坑湖的认识,将其作为一种复垦战略,努力减少尾矿水的环境足迹。
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引用次数: 0
Microbiological Treatment of Nitrate and Selenate from Coal Mine-Affected Water in a Subsurface,Semi-Passive, and in situ Water Treatment Facility 在地下半被动原位水处理设施中微生物处理煤矿影响水中硝酸盐和硒酸盐
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108259
Rachel Spietz, Lisa Kirk
Metallurgical coal mining generates significant amounts of waste rock, which can release nitrate and selenate upon atmospheric exposure, with resulting changes in surface water and ground water quality. Microorganisms residing on waste rock in the local subsurface have the metabolic potential to denitrifyand reduce selenium to treat mine-affected waters and stabilize waste rock. To support these processes, waste rock facilities have been designed to inject nutrients (e.g., methanol and phosphoric acid) in semi-passive water treatment of mine-impacted waters. Microbial community data regularly collected fromsubsurface semi-passive reactors throughout the startup and operations phases showed that native heterotrophic denitrifying bacteria and selenium reducing bacteria were enriched as a result of nutrient amendment. The microbial community was stable while the source water and geochemical parameters remained unchanged; however, a significant shift in the microbial community coincided with changes in the source of water treated. Metagenomic sequencing of microbial communities within the active treatment zone revealed multiple biochemical pathways of nitrate reduction. Geochemical and water quality data indicate near complete selenate reduction, yet a low abundance of in known selenate reduction genes were recovered. This may suggest that biologically mediated selenium reduction may be more widespread, both functionally and taxonomically. Further research into these pathways and mechanisms for nitrate and selenium reduction will help to strengthen our understanding of selenium reduction mechanisms and their application in mine water waste management.
冶金煤开采产生了大量的废石,这些废石暴露在大气中会释放硝酸盐和硒酸盐,从而导致地表水和地下水水质的变化。当地地下废石上的微生物具有反硝化和还原硒的代谢潜力,可以处理矿损水,稳定废石。为了支持这些过程,设计了废石设施,在受地雷影响的水的半被动水处理中注入营养物质(例如甲醇和磷酸)。在启动和运行阶段定期收集的地下半被动反应器微生物群落数据表明,由于营养修正,原生异养反硝化细菌和硒还原细菌得到了富集。在水源水和地球化学参数不变的情况下,微生物群落稳定;然而,微生物群落的显著变化与处理水源的变化相一致。活性处理区内微生物群落的宏基因组测序揭示了硝酸盐还原的多种生化途径。地球化学和水质数据表明硒酸盐几乎完全还原,但在已知的硒酸盐还原基因中恢复了低丰度。这可能表明生物介导的硒还原可能在功能和分类上更为广泛。进一步研究这些硝酸盐和硒的还原途径和机制,将有助于加强对硒还原机制的认识及其在矿山废水治理中的应用。
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引用次数: 0
Characterization of DNA Degrading Microorganisms from Dewar Creek Hot Springs in Western Canada 加拿大西部杜瓦溪温泉DNA降解微生物的研究
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108170
Sabina Pang, Triet Tran, Robert Bowers, Tanja Woyke, Peter Dunfield
Historically, discovery and subsequent characterization of microbial species relied on pure cultures. Some challenges associated with creating pure cultures have been overcome with advances in culture independent and DNA-based molecular methods such as single-cell genomics, metagenomics, or large scale amplicon sequencing. With these advances, the rate of discovery of new species from genomic data has quickly outpaced the number of organisms with cultured representatives. As description and characterization still rely on cultures, our understanding of yet uncultured species is greatly lacking. Major lineages in the bacterial domain equivalent to phyla that lack any cultured representatives are termed “candidate phyla.” Candidate phyla are found across the bacterial tree of life, and many uncultured organisms are found to be dominant in understudied environments. Extreme environments such as thermal springs are an example of understudied environments, making them excellent environments for studying novel microbial lineages. The objective of this research is to characterize uncultured bacterial lineages from Dewar Creek hot spring in Western Canada, with a focus on DNA and protein metabolizing bacteria. Based on previous genomic data from organisms in this hot spring, we hypothesize that the candidate phylum S2R-29, with extremely low GC content, metabolizes extracellular DNA or protein Dewar Creek is a thermal spring located in the Purcell Wilderness Conservancy in British Colombia, Western Canada. It is one of Canada’s hottest springs, reaching temperatures of up to 83˚C. Samples of DNA extracted from the Dewar Creek hot spring were PCR-amplified with primers for the v3v4 region of the 16S rRNA gene to detect S2R-29, and then sequenced on an Illumina Miseq. S2R-29 was found in samples ranging in temperature from 60˚C to 77 ˚C. In addition to samples from Dewar Creek, samples from other thermal springs in Canada, as well as samples from springs in New Zealand are also being sequenced with the same primers to determine the prevalence of this candidate phylum in other similar environments. This will give a better idea of the growth conditions and range of this organism. Previously, S2R-29 single amplified genomes (SAGs) were generated from Dewar Creek samples. Analysis of these SAGs suggests that S2R-29 has the potential to use peptides and DNA as carbon sources (Fig. 1). To test the potential to metabolize DNA and protein, enrichments of samples with 13C labelled dNTPs or 13C labelled protein have been started. These enrichments will be used for stable isotope probing (SIP) to determine if any organisms in Dewar Creek are metabolizing dNTPs or protein. In addition to SIP, primers specific for S2R-29 for quantitative PCR (qPCR) have been designed and will be run on the enrichments to determine if there are any changes in the abundance of S2R-29 over time in these enrichments, further testing the metabolic potential of these organisms. Finally, probe
从历史上看,微生物物种的发现和随后的表征依赖于纯培养。随着培养独立和基于dna的分子方法的进步,如单细胞基因组学、宏基因组学或大规模扩增子测序,与创造纯培养物相关的一些挑战已经被克服。随着这些进步,从基因组数据中发现新物种的速度已经迅速超过了具有培养代表的生物体的数量。由于描述和特征仍然依赖于文化,我们对尚未培养的物种的了解非常缺乏。在细菌领域的主要谱系相当于门,缺乏任何培养的代表被称为“候选门。”候选门在细菌生命树中被发现,许多未培养的生物被发现在未充分研究的环境中占主导地位。极端环境,如温泉,是一个未被充分研究的环境的例子,使它们成为研究新型微生物谱系的绝佳环境。本研究的目的是表征来自加拿大西部杜瓦溪温泉的未培养细菌谱系,重点是DNA和蛋白质代谢细菌。根据该温泉中生物的基因组数据,我们假设该候选门S2R-29代谢细胞外DNA或蛋白质,其GC含量极低,是位于加拿大西部不列颠哥伦比亚省Purcell Wilderness Conservancy的一个温泉。它是加拿大最热的温泉之一,最高温度可达83摄氏度。从杜瓦溪温泉提取DNA样本,用引物对16S rRNA基因的v3v4区进行pcr扩增,检测S2R-29,然后在Illumina Miseq上测序。S2R-29存在于温度范围为60℃至77℃的样品中。除了来自杜瓦溪的样本外,来自加拿大其他温泉的样本以及来自新西兰温泉的样本也正在用相同的引物进行测序,以确定该候选门在其他类似环境中的流行程度。这样可以更好地了解这种生物的生长条件和活动范围。此前,S2R-29单扩增基因组(SAGs)是从杜瓦溪样本中生成的。对这些sag的分析表明,S2R-29具有利用肽和DNA作为碳源的潜力(图1)。为了测试代谢DNA和蛋白质的潜力,已经开始用13C标记的dNTPs或13C标记的蛋白质富集样品。这些富集将用于稳定同位素探测(SIP),以确定杜瓦溪中是否有任何生物在代谢dNTPs或蛋白质。除了SIP之外,已经设计了用于定量PCR (qPCR)的S2R-29特异性引物,并将在富集物上运行,以确定这些富集物中S2R-29的丰度是否随时间变化,进一步测试这些生物的代谢潜力。最后,将为S2R-29设计荧光原位杂交(FISH)探针。这些将用于从杜瓦溪样品进行FISH,以便可视化这个候选门。通过这项研究,我们希望对这一新门进行表征,并可能发现杜瓦溪中DNA和蛋白质代谢细菌的其他新谱系。这项研究将有助于更好地理解这些未被充分研究的环境中的过程,并有助于我们理解细菌代谢在生物地球化学循环(如有机物的生物降解)中可能发挥的作用。总而言之,我们希望这项研究能够深入了解细菌为了在高温环境中茁壮成长而进化出的生活方式。
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引用次数: 0
Investigating Manganese-oxidizing microbial Biofilms in a historic Copper Mine of Upper Frankonia 法兰克尼亚上一历史铜矿中锰氧化微生物生物膜的研究
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108107
Tillmann Lueders, Christopher Mechela, Felix Beulig, Martin Obst
Providing evidence for presumed chemolithoautotrophic manganese oxidation remains a major and challenging objective in subsurface microbiology. Here, we report on the dissection of blackish, leathery microbial biofilms discovered in the “Goldene Falk”, a historic copper mine in Northern Bavaria, with mine shafts originating back to the 15 th centrury. Biogeochemical analysis of the biofilm indicated a notable enrichment of manganese oxides (MnOX), with Mn making up for more than 10% (dry weight) of the deposits. STXM analysis suggested a clear biogenic origin of MnOX in situ . Characteristic nodules of MnOX with microbial cells attached were also found in aerobic Mn-oxidizing enrichment cultures set up in minimal media in the lab. The biofilms obtained from the mine were also subjected to amplicon and metagenomic sequencing,revealing a vast diversity of presumably chemolithoautotrophic and heterotrophic microbial lineages, including members of the Pyrinomonadaceae , Rhizobiales , Methylomirabilaceae and also lineages within the Nitrospiraceae previously reported to be associated with lithotrophic Mn oxidation. We reconstructed >100 high-quality bacterial genomes (MAGs), many of them carrying genomic signatures of biogenic Mn oxidation (albeit non-lithotrophic). We continue to investigate the biofilms, our enrichment cultures and the metagenomic data obtained from the mine for further evidence of possible autotrophic manganese oxidation, the macroscopic leathery biofilm representing a likely habitat for these still enigmatic microbes. Indications for nitrogen and sulfur cycling also ongoing in the biofilms will also be discussed. This research contributes to a better understanding of the yet-enigmatic capacities of the microbiota in man-made subsurface environments.
在地下微生物学中,为假定的趋化岩自养锰氧化提供证据仍然是一个主要的和具有挑战性的目标。在这里,我们报道了在北巴伐利亚州一个历史悠久的铜矿“Goldene Falk”中发现的黑色皮革状微生物生物膜的解剖,该铜矿的矿井可以追溯到15世纪。生物地球化学分析表明,生物膜中锰氧化物(MnOX)显著富集,Mn占沉积物干重的10%以上。STXM分析表明,原位MnOX的生物源性很明显。在实验室中,在最小培养基中建立的好氧锰氧化富集培养中也发现了附着微生物细胞的MnOX特征结核。从该矿中获得的生物膜也进行了扩增子和宏基因组测序,揭示了可能是趋化岩石自养和异养的微生物谱系的巨大多样性,包括Pyrinomonadaceae, Rhizobiales, Methylomirabilaceae的成员,以及先前报道的与岩石营养Mn氧化有关的Nitrospiraceae中的谱系。我们重建了100个高质量的细菌基因组(MAGs),其中许多携带生物锰氧化的基因组特征(尽管非岩石营养)。我们继续研究生物膜,我们的富集培养和从矿山获得的宏基因组数据,以进一步证明可能的自养锰氧化,宏观皮革状生物膜代表了这些仍然神秘的微生物的可能栖息地。还将讨论生物膜中正在进行的氮和硫循环的适应症。这项研究有助于更好地理解微生物群在人造地下环境中的神秘能力。
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引用次数: 0
Prominence of Candidate Phyla Radiation (CPR) in Alberta Groundwater 阿尔伯塔省地下水中候选门辐射(CPR)的突出
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e110344
Damon Mosier
Groundwater is an essential part of everyday life, serving as agricultural irrigation, supporting numerous industrial processes, and providing drinking water for many. It is also home to a diverse range of unexplored microbial communities. This is especially true in Alberta, Canada, due to the rich geological history of the region and the close proximity of aquifer locations to areas ranging from oil sand-rich to agricultural to populous cities. Through collaborations with the Environment and Protected Areas division of the Alberta government we have access to more than 250 wells throughout the province, 25 of which were selected for metagenomics analysis. More than 750 metagenome-assembled genomes were recovered and coupled with historic geochemical, isotopic, and dissolved gas data, allowing us to interpret the lifestyles of microbial communities inhabiting Alberta groundwater. Results show a strong presence of organisms involved in C1-cycling, indicative of a productive subsurface environment, as well as members belonging to the Candidate Phyla Radiation (CPR). We explore likely roles of CPR, including their potential for ecological interactions via secondary metabolites. This research shows that Alberta groundwater is home to diverse, productive microbial communities that have the capacity to further our knowledge in microbial ecology.
地下水是日常生活的重要组成部分,用于农业灌溉,支持许多工业过程,并为许多人提供饮用水。它也是各种未知微生物群落的家园。在加拿大的阿尔伯塔省尤其如此,因为该地区有着丰富的地质历史,而且含水层位置靠近油砂丰富的地区、农业地区和人口稠密的城市。通过与艾伯塔省政府环境和保护区部门的合作,我们可以在全省范围内使用250多口井,其中25口被选中用于宏基因组学分析。超过750个宏基因组组装的基因组被恢复,并与历史地球化学、同位素和溶解气体数据相结合,使我们能够解释居住在阿尔伯塔省地下水中的微生物群落的生活方式。结果显示,参与c1循环的生物大量存在,表明存在生产性的地下环境,以及属于候选辐射门(CPR)的成员。我们探讨了CPR可能的作用,包括它们通过次生代谢物进行生态相互作用的潜力。这项研究表明,阿尔伯塔省地下水是多样化、多产的微生物群落的家园,这些微生物群落有能力进一步提高我们在微生物生态学方面的知识。
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引用次数: 0
The impact of subsurface life on ghost-rock karstification processes and cave formation 地下生命对鬼岩岩溶过程和洞穴形成的影响
Pub Date : 2023-10-17 DOI: 10.3897/aca.6.e108695
Guillaume Peugnet, Céline Pisapia, Laurent Bruxelles, Cédric Champollion, Philippe Vernant, Léna Lecourt, Bénédicte Ménèz, Emmanuelle Gérard
Karst systems represent an important carbon and freshwater reservoirs. Although karst systems have been studied for many years, a new paradigm has emerged that suggests some of them could be formed by ghost-rock processes (Dubois et al. 2014 ) . Contrarily to the classical total karstification, ghost-rock karstification leaves in place a weathered rock, called the ghost-rock, that can constitute a microbial habitat (Spilde et al. 2005). The first results of a geomicrobiological study of the Sterkfontein’s cave system in South Africa show that these ghost-rocks are mainly composed of iron and manganese oxides mixed with organic matter of putative microbial origin (Pisapia et al. in prep). To further understand the microbial community inhabiting these ghost-rocks, its specificity compared to groundwater, and its functional impact on the karst system of Sterkfontein, a metagenomic analysis from both ghost-rocks and groundwater samples was performed. It was completed by laser microdissection of the microorganisms attached to the mineral particles, followed by whole-genome amplification and transmission electron microscopy to analyze both the nature of the mineral particles and the microorganisms associated with them. The results highlight the differences in community between these two environments (with higher abundance of Actinobacteriota and Acidobacteriota in ghost-rock samples compared to ground water in particular), and suggest a high importance of microbe-minerals interactions in the ghost rocks, through metallophores production and extracellular electron transfer processes between bacteria and metallic ions.
喀斯特系统是重要的碳和淡水储集层。尽管对喀斯特系统的研究已经进行了多年,但一种新的范式已经出现,认为其中一些喀斯特系统可能是由鬼岩过程形成的(Dubois et al. 2014)。与经典的全岩溶作用相反,鬼岩岩溶作用留下了风化的岩石,称为鬼岩,可以构成微生物栖息地(Spilde et al. 2005)。对南非Sterkfontein洞穴系统进行的地球微生物学研究的初步结果表明,这些鬼岩主要由铁和锰的氧化物组成,并混合了假定的微生物来源的有机物(Pisapia等人)。为了进一步了解这些鬼岩中的微生物群落、其与地下水的特异性及其对Sterkfontein岩溶系统的功能影响,对鬼岩和地下水样品进行了宏基因组分析。通过激光显微解剖附着在矿物颗粒上的微生物,然后进行全基因组扩增和透射电子显微镜来分析矿物颗粒及其相关微生物的性质。结果突出了这两种环境之间的群落差异(特别是与地下水相比,鬼岩样品中放线菌群和酸杆菌群的丰度更高),并表明鬼岩中微生物-矿物相互作用的高度重要性,通过金属基团的产生和细菌与金属离子之间的细胞外电子转移过程。
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引用次数: 0
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