Did mineral surface chemistry and toxicity contribute to evolution of microbial extracellular polymeric substances?

Abstract Modern ecological niches are teeming with an astonishing diversity of microbial life in biofilms closely associated with mineral surfaces, which highlights the remarkable success of microorganisms in conquering the challenges and capitalizing on the benefits presented by the mineral-water interface. Biofilm formation capability likely evolved on early Earth because biofilms provide crucial cell survival functions. The potential toxicity of mineral surfaces toward cells and the complexities of the mineral-water-cell interface in determining the toxicity mechanisms, however, have not been fully appreciated. Here, we report a previously unrecognized role for extracellular polymeric substances (EPS), which form biofilms in shielding cells against the toxicity of mineral surfaces. Using colony plating and LIVE/DEAD staining methods in oxide suspensions versus oxide-free controls, we found greater viability of wild-type, EPS-producing strains of Pseudomonas aeruginosa PAO1 compared to their isogenic knockout mutant with defective biofilm-producing capacity. Oxide toxicity was specific to its surface charge and particle size. High resolution transmission electron microscopy (HRTEM) images and assays for highly reactive oxygen species (hROS) on mineral surfaces suggested that EPS shield via both physical and chemical mechanisms. Intriguingly, qualitative as well as quantitative measures of EPS production showed that toxic minerals induced EPS production in bacteria. By determining the specific toxicity mechanisms, we provide insight into the potential impact of mineral surfaces in promoting increased complexity of cell surfaces, including EPS and biofilm formation, on early Earth. Key Words: Mineral toxicity-Bacteria-EPS evolution-Biofilms-Cytotoxicity-Silica-Anatase-Alumina. Astrobiology 12, 785-798.     

The simulated silicification of bacteria--new clues to the modes and timing of bacterial preservation and implications for the search for extraterrestrial microfossils

Evidence of microbial life on Earth has been found in siliceous rock formations throughout the geological and fossil record. To understand the mechanisms of silicification and thus improve our search patterns for evidence of fossil microbial life in rocks, a series of controlled laboratory experiments were designed to simulate the silicification of microorganisms. The bacterial strains Pseudomonas fluorescens and Desulphovibrio indonensis were exposed to silicifying media. The experiments were designed to determine how exposure time to silicifying solutions and to silicifying solutions of different Si concentration affect the fossilization of microbial biofilms. The silicified biofilms were analyzed using transmission electron microscopy (TEM) in combination with energy-dispersive spectroscopy. Both bacterial species showed evidence of silicification after 24 h in 1,000 ppm silica solution, although D. indonensis was less prone to silicification. The degree of silicification of individual cells of the same sample varied, though such variations decreased with increasing exposure time. High Si concentration resulted in better preservation of cellular detail; the Si concentration was more important than the duration in Si solution. Even though no evidence of amorphous silica precipitation was observed, bacterial cells became permineralized. High-resolution TEM analysis revealed nanometer-sized crystallites characterized by lattice fringe-spacings that match the (10-11) d-spacing of quartz formed within bacterial cell walls after 1 week in 5,000 ppm silica solution. The mechanisms of silicification under controlled laboratory conditions and the implication for silicification in natural environments are discussed, along with the relevance of our findings in the search for early life on Earth and extraterrestrial life.     

Microbially induced iron precipitation associated with a neutrophilic spring at Borra Caves, Vishakhapatnam, India

The present investigation uncovers various pieces of evidence for the possible biologically induced mineralization in iron mats associated with a pH-neutral spring in the Borra caves, Vishakhapatnam, India. Electron microscopy [scanning electron microscopy (SEM) and transmission electron microscopy (TEM)] demonstrated large numbers of (i) hollow tubes (diameter ～1?μm) resembling sheaths of the iron-oxidizing bacteria Leptothrix, (ii) thin (diameter <1?μm) solid fibers of uncertain origin, (iii) nanoscale subspherical to irregularly shaped particles encrusting tubes and fibers, and (iv) aggregates of broken and partially disintegrated sheaths, fibers, and particles embedded in extracellular polymeric substances (EPS) occasionally including microbial cells. X-ray microanalyses by energy dispersive spectroscopy (EDS) revealed that the mat accumulated largely Fe but also smaller amounts of Si and traces of P and Ca. Particles rich in Si and Al (possibly kaolinite) and Ca (carbonate) were also observed. High-resolution TEM/EDS of unstained ultrathin sections suggests that microbial sheaths were highly mineralized by amorphous to cryptocrystalline Fe-rich phases and less frequently by other fine-grained and fibrous authigenic claylike minerals. Total number of microorganisms in the iron mats was 5.8×10(5) cells, g sed(-1) (wet weight). Analysis of the 16S rRNA gene diversity revealed microorganisms assigned to eight different phyla [Proteobacteria (62%), Chloroflexi (8%), Bacteroidetes (7%), Planctomycetes (1%), Actinobacteria (5%), Acidobacteria (6%), Nitrospira (1%), Firmicutes (5%)]. Within the Proteobacteria, Betaproteobacteria was the predominant class, which accounted for 28% of the sequences. Within this class some obvious similarities between the obtained sequences and sequences from other cave systems could be seen, especially sequences affiliated with Leptothrix, Siderooxidans, Crenothrix, Comamonadaceae, Dechloromonas, and many uncultured Betaproteobacteria. Four (4%) of the sequences could not be assigned to phylum level but were affiliating with the candidate division TM7 (2%), candidate division OP11 (1%), and candidate division WWE3 (1%). The results allow us to infer a possible relationship of microbial sheaths, EPS, and the iron precipitates to microbial community diversity in the Borra cave springs. Understanding biogenic iron oxides in caves has important astrobiological applications as it provides a potential tool for the detection of extraterrestrial life.     

含磷聚酰亚胺薄膜在原子氧环境中的降解研究

文章考察了自行合成的含磷聚酰亚胺（PI）薄膜在模拟原子氧环境中的降解行为。扫描电子显微镜（SEM）、X射线光电子能谱（XPS）等测试结果表明:在原子氧辐照过程中,含磷PI薄膜表面的磷元素与氧元素含量增加,原子结合能也增大,意味着在PI表面形成了含磷钝化层。该钝化层进一步阻止了PI次表面层被侵蚀,使含磷PI薄膜表现出了抗原子氧侵蚀能力,其在模拟原子氧环境中的质量损失率远低于Kapton薄膜。     

Rock surfaces as life indicators: new ways to demonstrate life and traces of former life

Life and its former traces can only be detected from space when they are abundant and exposed to the planetary atmosphere at the moment of investigation by orbiters. Exposed rock surfaces present a multifractal labyrinth of niches for microbial life. Based upon our studies of highly stress-resistant microcolonial fungi of stone monument and desert rock surfaces, we propose that microbial biofilms that develop and become preserved on rock surfaces can be identified remotely by the following characteristics: (1) the existence of spectroscopically identifiable compounds that display unique adsorption, diffraction, and reflection patterns characteristic of biogenerated organic compounds (e.g., chlorophylls, carotenes, melanins, and possibly mycosporines), (2) demonstrably biogenic geomorphological features (e.g., biopitting, biochipping, and bioexfoliation), and (3) biominerals produced in association with biofilms that occupy rock surfaces (e.g., oxalates, forsterite, and special types of carbonates, sulfides, and silicates). Such traces or biosignatures of former life could provide macroscopically visible morphotypes and chemically identifiable products uniquely indicative of life.     

Chemical mapping of proterozoic organic matter at submicron spatial resolution

A NanoSIMS ion microprobe was used to map the submicron-scale distributions of carbon, nitrogen, sulfur, silicon, and oxygen in organic microfossils and laminae in a thin section of the approximately 0.85 billion year old Bitter Springs Formation of Australia. The data provide clues about the original chemistry of the microfossils, the silicification process, and the biosignatures of specific microorganisms and microbial communities. Chemical maps of fossil unicells and filaments revealed distinct wall- and sheath-like structures enriched in C, N, and S, consistent with their accepted biological origin. Surprisingly, organic laminae, previously considered to be amorphous, also exhibited filamentous and apparently compressed spheroidal structures defined by strong enrichments in C, N, and S. By analogy to NanoSIMS data from the well-preserved microfossils, these structures were interpreted as being of biological origin, most likely representing densely packed remnants of microbial mats. Given that the preponderance of organic matter in Precambrian sediments is similarly "amorphous," our findings indicate that a re-evaluation of ancient specimens via in situ structural, chemical, and isotopic study is warranted. Our analyses have led us to propose new criteria for assessing the biogenicity of problematic kerogenous materials, and, thus, these criteria can be applied to assessments of poorly preserved or fragmentary organic residues in early Archean sediments and any that might occur in meteorites or other extraterrestrial samples.     

Experimental hypervelocity impacts: Implication for the analysis of material retrieved after exposure to space environment: Part I. Impacts on aluminium targets

During the last three decades a wide variety of surfaces have been brought back to Earth after being exposed to space environment. The impact features found on these surfaces are used to evaluate the damages caused to spacecraft and can give clues to the characteristics of the orbital debris and meteoroids that created them. In order to derive more precisely the particle parameters and to improve the analysis of projectile remnants, we have performed an extensive analysis of craters caused by the impact of high velocity particles on thick ductile targets, using a micro-particle accelerator. We show that from the geometry of the craters and from the analysis of the remnants it is possible to derive the main characteristics of the projectiles. In particular, using up-to-date instrumentation, scanning electron microscope (SEM) and Energy Dispersive X-ray (EDX) spectrometer, we found that even small residues inside craters can be identified. However, this study shows that a velocity resolution better than 1 km/s would be appropriate to obtain a fair calibration of the impact processes on a ductile target. This would allow to decipher with precision impact features on ductile surfaces exposed to space environment.     

A transmission electron microscopy study of silica and kerogen biosignatures in approximately 1.9 Ga gunflint microfossils

Microfossils preserved in chert from the;1.9 Ga Gunflint Formation (Schreiber Beach, Ontario, Canada) were studied with transmission electron microscopy (TEM) and analytical TEM (ATEM). Our goals were to uncover the style of silicification relative to the distribution of organic matter, and to evaluate the distribution and evolution of organic matter, at submicroscopic spatial scales. Petrographically the microfossils typically display filamentous or coccoidal morphologies, and consist of quartz crystals surrounded by kerogen along grain boundaries. ATEM analysis revealed that quartz associated with kerogen consists of 200-500nm-sized, round crystallites, whereas the chert matrix is comprised of randomly oriented, polygonal microquartz (5-10 microm). Silica spheroids found within some fossils consist of quartz subgrains in an amorphous to poorly crystalline matrix, suggesting that precipitation of opaline silica on organic matter occurred with subsequent but incomplete transformation to quartz. Some coccoidal microfossils surround large euhedral quartz crystals (up to 5 microm in diameter) that appeared to have influenced the distribution of kerogen during crystal growth. These euhedral quartz crystals commonly contain elongated (50-100 nm) iron-rich crystallites. Energy-loss, near-edge structure analysis of kerogen associated with a coccoidal microfossil showed that it is composed of amorphous carbon with no evidence of graphitization. TEM results revealed significant differences in the style of silicification between microbe-shaped microfossils and their surrounding chert matrix, as well as the presence of amorphous kerogen.     

热循环对原子氧与树脂基复合材料 交互作用的影响

文章选取M40/DFA-1和M40/AG-80两种碳纤维增强树脂基复合材料,进行了热循环对原子氧与这两种材料交互作用影响的研究。原子氧反应率的计算和层间剪切性能测试的结果表明,一定温度区间一定次数的热循环使得材料与原子氧交互作用减弱。该研究应用X射线光电子能谱仪（XPS）和扫描电子显微镜（SEM）对样品表面成分和形貌进行了分析。根据分析结果,初步提出热循环对增强树脂基复合材料自身抗原子氧侵蚀能力的作用机理。     

Potential of cathodoluminescence microscopy and spectroscopy for the detection of prokaryotic cells on minerals

Detecting mineral-hosted ecosystems to assess the extent and functioning of the biosphere from the surface to deep Earth requires appropriate techniques that provide, beyond the morphological criteria, indubitable clues of the presence of prokaryotic cells. Here, we evaluate the capability of cathodoluminescence microscopy and spectroscopy, implemented on a scanning electron microscope, to identify prokaryotes on mineral surfaces. For this purpose, we used, as a first step, a simple model of either unstained or stained cultivable cells (Escherichia coli, Deinococcus radiodurans) deposited on minerals that are common in the oceanic crust (basaltic glass, amphibole, pyroxene, and magnetite). Our results demonstrate that the detection of cells is possible at the micrometric level on the investigated minerals through the intrinsic fluorescence of their constituting macromolecules (aromatic amino and nucleic acids, coenzymes). This allows us to distinguish biomorph inorganic phases from cells. This easily implemented technique permits an exploration of colonized rock samples. In addition, the range of spectrometric techniques available on a scanning electron microscope can provide additional information on the nature and chemistry of the associated mineral phases, which would lead to a simultaneous characterization of cells, their microhabitats, and a better understanding of their potential relationships.     

Kapton抗原子氧侵蚀的Al2O3涂层研究

在原子氧侵蚀地面模拟设备中对Kapton和利用反应溅射制备的Al2O3涂层进行了原子氧暴露实验，并采用XPS和SEM等分析手段对暴露前后试样表面的物理和化学变化进行了研究。结果表明，Kapton试样遭受了严重的侵蚀，质量损失较大；Al2O3涂层质量变化很小，对基体提供了良好的保护作用。XPS分析结果表明，Kapton的羰基与原子氧作用时形成CO2，随后CO2气体脱附。反应溅射的Al2O3涂层是富Al的，初始暴露时由于氧化反应而质量有少许增加，随时间延长，涂层变得完全符合化学计量。     

K42O2高温合金激光选区熔化成形微观组织研究

为了实现激光选区熔化成形(SLM)这项新工艺在液体火箭发动机高温合金结构上的推广应用,明晰其强化机理以及研究相应的热处理制度,对激光选区熔化快速凝固条件下组织形成及演化规律、第二相析出特点进行了分析和讨论.采用SLM成形K4202镍基高温合金试样,通过光学显微镜(OM)、扫描电镜(SEM)及X射线衍射(XRD)等理化分析手段,将基于ANSYS生死单元技术的温度场数值模拟结果和经典凝固理论相结合,揭示了其组织特征及演化规律:沉积态表现为外延生长柱状晶,层间可见层带组织,顶部出现转向枝晶和二次枝晶臂,γ'强化相和碳化物的析出受到抑制.     

防静电Kapton二次表面镜的电子辐照效应

文章研究防静电Kapton二次表面镜（ITO/Kapton/Al）在不同能量电子辐照后,太阳吸收比退化情况,并利用X射线光电子能谱仪（XPS）和扫描电子显微镜（SEM）对样品表面的成分和形貌进行了分析。结果表明,相同电子注量下,电子能量越高,对样品的太阳吸收率退化作用越大;电子辐照对样品表面形貌影响不大,但是会降低表面氧化铟锡的含量;不同能量的电子对样品表面成分的损伤效应是一致的。     

片状Al及Al//Mg合金粉的制备及性能

针对金属/水推进剂对金属粉的要求,采用双向旋转球磨机制备了片状Al及Al/Mg合金粉.应用扫描电子显微镜(SEM)、X射线粉末衍射仪(XRD)、差示扫描量热仪(TG-DSC)、简易点火温度和水反应特性测试装置等手段分析了产物的形貌、结构及高温氧化过程、点火温度和水反应特性等.与球形Al对比结果表明,片状Al及Al/Mg合金粉总增重率大幅提高,初始放热峰温降低,活性增强,点火温度分别降低150℃和400 ℃以上.Al/Mg合金粉的水反应活性优于片状Al粉.     

细菌生物膜对航天器材料的危害及其防护简述

细菌生物膜是造成空间环境下航天器材料腐蚀的关键因素之一.文章简述了细菌生物膜的结构和形成机制、其对航天器材料的危害、主要评估方法以及防控策略,并提出目前防控策略在实际应用中存在的问题.当前的细菌生物膜防控策略主要分为3类:1)修改易感表面的特性以防止细菌生物膜形成;2)调节信号通路以抑制细菌生物膜的形成;3)施加外力以...     

With a grain of salt: what halite has to offer to discussions on the origin of life

This experimental study investigated how the dynamics of the crystallization of the evaporite mineral halite could affect the accumulation and preservation of organic macromolecules present in the crystallizing solution. Halite was grown under controlled conditions in the presence of polymer nanoparticles that acted as an analog to protocellular material. Optical microscopy, atomic force microscopy, and laser scanning confocal fluorescence microscopy were used to trace the localization of the nanoparticles during and after growth of halite crystals. The present study revealed that the organic nanoparticles were not regularly incorporated within the halite, but were very concentrated on its surfaces. Their distribution was controlled dominantly by the morphologic surface features of the mineral rather than by specific molecular interactions with an atomic plane of the mineral. This means that the distribution of organic molecules was controlled by surfaces like those of halite's evaporitic growth forms. The experiments with halite also demonstrated that a mineral need not continuously incorporate organic molecules during its crystallization to preserve those molecules: After rejection by (non-incorporation into) the crystallizing halite, the organic nanoparticles increased in concentration in the evaporating brine. They ultimately either adsorbed in rectilinear patterns onto the hopper-enhanced surfaces and along discontinuities within the crystals, or they were encapsulated within fluid inclusions. Of additional importance in origin-of-life considerations is the fact that halite in the natural environment rapidly can change its role from that of a protective repository (in the absence of water) to that of a source of organic particles (as soon as water is present) when the mineral dissolves.     

Calcification and silicification: fossilization potential of cyanobacteria from stromatolites of Niuafo'ou's Caldera Lakes (Tonga) and implications for the early fossil record

Calcification and silicification processes of cyanobacterial mats that form stromatolites in two caldera lakes of Niuafo'ou Island (Vai Lahi and Vai Si'i) were evaluated, and their importance as analogues for interpreting the early fossil record are discussed. It has been shown that the potential for morphological preservation of Niuafo'ou cyanobacteria is highly dependent on the timing and type of mineral phase involved in the fossilization process. Four main modes of mineralization of cyanobacteria organic parts have been recognized: (i) primary early postmortem calcification by aragonite nanograins that transform quickly into larger needle-like crystals and almost totally destroy the cellular structures, (ii) primary early postmortem silicification of almost intact cyanobacterial cells that leave a record of spectacularly well-preserved cellular structures, (iii) replacement by silica of primary aragonite that has already recrystallized and obliterated the cellular structures, (iv) occasional replacement of primary aragonite precipitated in the mucopolysaccharide sheaths and extracellular polymeric substances by Al-Mg-Fe silicates. These observations suggest that the extremely scarce earliest fossil record may, in part, be the result of (a) secondary replacement by silica of primary carbonate minerals (aragonite, calcite, siderite), which, due to recrystallization, had already annihilated the cellular morphology of the mineralized microbiota or (b) relatively late primary silicification of already highly degraded and no longer morphologically identifiable microbial remains.     

不同截面炭纤维表面特性及其对复合材料界面粘接性能的影响

使用SEM、AFM、比表面积与孔体积测试分析仪(BET)和XPS对国产腰形截面炭纤维、圆形截面高强炭纤维和国外圆形截面T300炭纤维表面特性进行物理与化学表征与分析,并对炭纤维/环氧复合材料界面粘接性能进行了研究。表面形貌分析表明,腰形截面炭纤维比表面积大于圆形截面炭纤维,但其表面沟槽较圆形截面炭纤维浅。XPS分析表明,腰形截面炭纤维的表面活性略高于国产圆形截面炭纤维,但明显低于T300;界面剪切强度与层间剪切强度测试结果表明,腰形截面炭纤维/环氧复合材料的界面剪切强度和层间剪切强度均接近于T300/环氧复合材料,高于国产圆形截面炭纤维/环氧复合材料。     

预分散酚醛中空微球对三元乙丙橡胶绝热层性能的影响及梯度化绝热层的研究

分别制备了预分散酚醛中空微球填充三元乙丙橡胶绝热层和梯度化三元乙丙橡胶绝热层,旨在开发低密度的新型高性能绝热材料。扫描电镜观察表明,预分散酚醛中空微球能完好、均匀地分散于三元乙丙橡胶基体中。由于酚醛中空微球具有很低的粒子密度和高的热阻性能,随预分散酚醛中空微球含量的增加,绝热层的密度、热导率和热扩散率显著降低,同时绝热层的比热容明显增大,表现出优异的绝热性能。与芳砜纶浆粕填充三元乙丙橡胶绝热层、酚醛中空微球填充三元乙丙橡胶绝热层相比,梯度化三元乙丙橡胶绝热层具有优异的绝热性能和耐烧蚀性能,同时又具有很低的密度,将在火箭发动机上具有广阔的应用前景。     

K4202镍基高温合金激光选区熔化成形室温拉伸性能研究

为了将激光选区熔化(SLM)这项技术推广到液体火箭发动机高温合金复杂结构件的成形,满足其使用要求,对SLM成形K4202高温合金力学性能及其强化机理进行研究。沉积态室温下拉伸试验力学性能指标表现出了很强的各向异性,但均接近或超过GH4202锻件标准值,通过光学显微镜(OM)、扫描电镜(SEM)、X射线衍射(XRD)、透射电镜(TEM)等理化分析手段揭示了其强化机理主要为细晶强化、应变硬化、沉淀硬化和过饱和的固溶强化。同时研究了固溶、固溶时效、直接时效三种热处理制度对K4202力学性能的影响,结果表明直接时效后的综合力学性能最佳。