Distribution and Diversity of Aerobic Carbon Monoxide-Oxidizing Bacteria in Geothermal Springs of China,the Philippines,and the United States

Accumulating genomic evidence suggests that a variety of thermophilic bacteria contain cox operons and may be capable of aerobic carbon monoxide (CO) oxidation. However, little is known about the distribution and diversity of the cox-encoding (COXE) bacteria in natural geothermal environments. In this study, we examined coxL gene (encoding the large subunit of carbon monoxide dehydrogenase: CoxL) sequences retrieved from the sediments of 25 geothermal sites located in the Qinghai-Tibetan Plateau (QTP) and Yunnan Province (YP) of China, the Bacon-Manito Geothermal Production Field (BGPF) of the Philippines, and the Great Basin of the United States (USGB). Temperature and pH ranges of the studied hot springs were 22.1 to 90.8°C and 2.7 to 9.4, respectively. Phylogenetic analyses showed that most CoxL sequences were closely related to the classes Actinobacteria, Deinococci, Ktedonobacteria, Thermomicrobia, and Clostridia, and hot springs from different regions hosted different COXE communities. In addition, these hot springs harbored some COXE bacteria that were phylogenetically distinct from those inhabiting nongeothermal ecosystems. This study revealed no significant correlation between temperature or pH and the composition or diversity of COXE communities at the global scale. However, within a given region, temperature was correlated with the COXE bacterial community composition.     

云南地区热泉中氨氧化菌丰度对环境条件的响应

【目的】研究热泉中的氨氧化菌对于理解全球氮循环作用至关重要,而人们对于热泉中环境条件对氨氧化菌丰度分布的影响还知之甚少。本研究旨在研究云南热泉中氨氧化菌的丰度以及热泉环境因子(例如:温度、氨浓度及pH等)对氨氧化菌丰度的影响。【方法】在所选取的热泉中,采集沉积物、菌席或泉华样品。使用RNA逆转录、定量聚合酶链式反应及荧光原位杂交等技术对样品中各微生物种群进行定量分析。【结果】所选取的热泉沉积物、菌席或泉华中微生物总量大约为108-109细胞/g。其中,氨氧化古菌(AOA)占样品中微生物总量的0.02-1.32%,而氨氧化细菌数量低于检测下限。地球化学参数和AOA相对丰度的相关性统计分析显示,氨氧化古菌相对丰度值与NH3、NO2-、NO3-浓度和温度等具有统计学意义上的相关性,而其与Fe2+和及盐度无统计学意义上的相关性。【结论】在所调查的热泉中,氨氧化微生物种群主要由AOA组成,AOA在热泉中的氨氧化生物地球化学过程中起着重要作用。热泉中多个环境因子一起控制着AOA丰度在不同热泉中的分布特征,而某些环境因子,如盐度-和Fe2+浓度,可能不是控制AOA分布特征的关键因素。     

Actinobacterial Diversity in Microbial Mats of Five Hot Springs in Central and Central-Eastern Tibet,China

The diversity and community composition of Actinobacteria in microbial mats of five Tibetan hot springs (temperatures 26°C to 81°C) and a sympatric soil were investigated with 16S rRNA gene phylogentic analysis. A total of 278 clones were obtained. The actinobacterial communities in the Tibetan hot springs were diverse, and most of the retrieved clones were affiliated with Actinobacteridae, Acidimicrobidae, and unclassified Actinobacteria. The Actinobacteridae sequences were distributed into seven suborders (e.g., Frankineae, Corynebacterineae, Micromonosporineae, Pseudonocardineae, Propionibacterineae, Micrococcineae, and Actinomycineae) and unclassified Actinobacteridae. The actinobacterial composition varied among different hot springs. Statistical analysis showed that the actinobacterial diversity in the investigated Tibetan hot springs was not significantly correlated with temperature, suggesting that temperature is not a key factor in shaping the actinobacterial diversity in hot springs.     

Actinobacterial Diversity in Hot Springs in Tengchong (China), Kamchatka (Russia), and Nevada (USA)

Actinobacterial diversity was investigated in three hot springs [Tengchong Frog Mouth (TFM) hot spring (in Tengchong, China), Kamchatka Robb Flag hot spring (in Kamchatka, Russia), and Nevada Boiling Spring (in Nevada, USA)], a Tengchong Frog Mouth hot spring-derived pond and a soil sample near the TFM hot spring. Culture-independent techniques were employed including denaturing gradient gel electrophoresis and actinobacterial 16S rRNA gene phylogenetic analysis. The three hot springs and the hot spring-derived pond had pH of 6.8 to 7.9 and temperature of 50 to 81°C. Actinobacteria were for the first time found to be very diverse in hot springs with temperature up to 81°C, indicating that they have an unexpected capability of adapting to hot spring environments. A total of 346 actinobacterial clones were obtained from the three hot springs, and these clones were identified as 31 RFLP types, falling into the Actinomycetales (only limit to the Frankineae suborder), the Rubrobacterales, uncultured Candidatus Microthrix, unclassified Actinobacteria, and uncultured Candidate Division OP10. Our data have implications for a better understanding of the distribution of Actinobacteria in hot springs.     

Lipid Biomarkers,Carbon Isotopes,and Phylogenetic Characterization of Bacteria in California and Nevada Hot Springs

Microbial mats were collected from hot springs in California (Eagleville) and Nevada (Paradise Valley and Crescent Valley) to determine bacterial community structure and pathways of carbon cycling in different geothermal environments of the western United States. Phospholipid fatty acids (PLFA) at Eagleville contained even-numbered fatty acids, with 16:0 being the most abundant (48.8%), followed by 18:1ω 9c (17.2%), 16:1ω 7c/t (6.3%), and 18:0 (6.2%), which are consistent with lipid profiles of cyanobacteria or other phototrophic bacteria. The PLFA profiles at Paradise Valley and Crescent Valley were dominated by similar even-numbered fatty acids; however, branched fatty acids such as iso- and anteiso- 15:0 and 17:0 were also abundant (up to 7.1% compared to 2.0% at Eagleville), suggesting greater relative abundance of heterotrophic bacteria in these springs. Analysis of neutral lipids was only performed on Eagleville and Paradise Valley springs, which revealed abundant bacterial hopanoids including the 2–methylbacteriohopane-32,33,34,35-tetrol (2-methylBHT) that is specific to cyanobacteria; however, the diversity of hopanoid compounds was significantly lower at Eagleville than at Paradise Valley. The carbon-isotope composition of individual PLFA averaged ?30.7 ± 1.3‰ (n = 7) at Eagleville, ?28.0 ± 1.8‰ (n = 3) at Crescent Valley, and ?29.7 ± 3.1‰ (n = 12) at Paradise Valley. Carbon isotope fractionation between PLFA and CO ₂ was only available for Eagleville (?11.7‰) and Paradise Valley (?21.7‰), which indicated the predominance of the Calvin cycle for CO ₂ fixation in these hot springs. Bacterial 16S rRNA genes were extracted from environmental samples at Eagleville and Paradise Valley but not Crescent Valley. Clone libraries indicated the predominance of cyanobacteria (50–75%) at these locations, which is consistent with the lipid profiles. Phylogenetic tree of the 16S rRNA genes indicated that most of the cyanobacterial sequences are unknown and may be specific to the Nevada and California hot springs. Phototrophic green non-sulfur bacteria were also present at Eagleville (13%) and Paradise Valley (7%). The remaining sequences were related to α-, β -, and γ -Proteobacteria, Acidobacteria, Deinococcus/Thermus, Bacteroidetes, and Spirochaetes. However, not all of these sequences were present at each of the springs. Results of this study demonstrate the consistency among lipid profiles (phenotypes), carbon isotopes (biogeochemistry), and 16S rRNA genes (genotypes) of the bacterial community in these hot springs, which cumulatively suggest the importance of cyanobacteria in primary production of biomass under the environmental conditions examined.     

Microbial Silicification in Sinters from Two Terrestrial Hot Springs in the Uzon Caldera,Kamchatka, Russia

Silicification of microbial communities is the most pervasive form of mineralization in two terrestrial hot springs in the Uzon Caldera. X-ray diffraction and electron microscopy reveal a diverse sinter mineral assemblage dominated by opal-A with accessory sulfur, sulfides, alunite group minerals, oxides, and oxyhydroxides. Aluminum laminations (reported for the first time) noted in one deposit may slow opal-A transformational rates enabling mineralized microbial remnants to exist longer in the rock record. Although preservation of microbial forms decreases over time, the collective mineral assemblage and patterning suggests that they are the most persistent lines of evidence of life in the geologic record.     

Abundance and Diversity of Ammonia-Oxidizing Bacteria and Archaea in Cold Springs on the Qinghai-Tibet Plateau

The cold springs underlain by gas hydrates on the Qinghai-Tibet Plateau (QTP) are similar to deep-sea cold seeps with respect to methane biogeochemistry. Previous studies have shown that ammonia oxidizing bacteria (AOB) and archaea (AOA) are actively present and play important roles in the carbon/nitrogen cycles in cold seeps. Studying AOA and AOB communities in the QTP cold springs will be of great importance to our understanding of carbon and nitrogen cycling dynamics related to the underlying gas hydrates on the QTP. Thus, the abundance and diversity of AOB and AOA in sediments of four cold springs underlain by gas hydrates on the QTP were determined by using quantitative polymerase chain reaction and amoA gene (encoding ammonia monooxygenase involved in ammonia oxidation) phylogenetic analysis. The results showed that the AOB and AOA amoA gene abundances were at 10³–10⁴ copies per gram of the sediments in the investigated cold springs. The AOB population consisted of Nitrosospira and Nitrosomonas in contrast with the mere presence of Nitrosospira in marine cold seeps. The AOB diversity was higher in cold springs than in cold seeps. The AOA population was mainly composed of Nitrososphaera, in contrast with the dominance of Nitrosopumilus in cold seeps. The terrestrial origin and high level of dissolved oxygen of the cold springs may be the main factors accounting for the observed differences in AOB and AOA populations between the QTP cold springs and marine cold seeps.     

Isolation and Characterization of Thermophilic Bacteria from Gavmesh Goli Hot Spring in Sabalan Geothermal Field,Iran: Thermomonas hydrothermalis and Bacillus altitudinis Isolates as a Potential Source of Thermostable Protease

Abstract

Thermophilic bacteria have attracted great attention due to their ability to produce thermostable enzymes. The aim of this study was the isolation and characterization of thermophilic bacteria from Gavmesh Goli hot spring in Sareyn, North West of Iran. Of 10 water samples collected, 36 thermophilic bacteria were obtained. The thermophilic bacteria were tested for their ability to produce hydrolase enzymes. All the isolates were potentially protease producers. Lipase, DNase, and amylase activities were confirmed in 34 (94.4%), 8 (22.2%), and 3 (8.3%) isolates, respectively. Five isolates with higher levels of enzyme activity were selected for further studies. Morphological, biochemical, and molecular analysis by 16S rRNA gene sequencing revealed that four isolates (DH15, DH16, DH20, and DH29) could be identified as Thermomonas hydrothermalis and one (PA10) Bacillus altitudinis. The protease produced by these isolates was optimally active at 50–55?°C, pH 8–8.5, and 0–0.5?M NaCl. In this first time study, we isolated T. hydrothermalis and B. altitudinis from Iranian hot springs and demonstrated the characteristics of T. hydrothermalis protease. Accordingly, due to the valuable potential of these bacteria such as the production of protease with high temperature and pH stability, these isolates can be introduced as promising candidates for industrial applications.     

滇藏热泉arxA基因型厌氧砷氧化原核微生物多样性及其影响因素

【目的】探究滇藏地区热泉沉积物微生物及arxA基因的多样性及其影响因素。【方法】采用Illumina MiSeq高通量测序技术和统计学的分析方法,针对3个滇藏地热区的22个样点,进行地球化学分析及微生物多样性分析,并分析其与环境因子之间的相关性。【结果】沉积物微生物群落主要门类及其丰度占比为Chloroflexi(21.27%)、Deinococcus-Thermus(17.25%)、Aquificae(13.39%)、Proteobacteria(9.27%)、Acetothermia(8.3%)、Bacteroidetes(4.96%)与Crenarchaeota(4.57%);具有arxA基因的微生物主要门类及其丰度占比分别为Proteobacteria(64.87%)、Bipolaricaulota (9.55%)、Deinococcus-Thermus (6.42%)与Crenarchaeota (4.05%);不同地热区优势种群有所差异,主要受温度、pH值、砷浓度、总溶解固体(TDS)及海拔等因素的影响,Mantel检验呈显著相关性(P<0.001)。【结论】沉积物中...     

Microbial Mat Boundaries between Chemolithotrophs and Phototrophs in Geothermal Hot Spring Effluents

Among the various microbial mats that develop in geothermal hot springs in solfataric fields, colorless sulfur-turf (ST)—macroscopic white bundles consisting of large sickle-shaped bacteria belonging to Aquificales and elemental sulfur particles–develops in a limited environment of geothermal effluent containing hydrogen sulfide with neutral pH and low in oxygen. Photosynthetic cyanobacterial mat (CY) often grow just downstream of chemolithotrophic ST, or they coexist with ST where the temperature is slightly lower. Knowledge of the environmental regimes of these microbial mats will lead to better understanding of the distribution of thermophilic microorganisms on the Earth and provide clues about evolutionary processes in the microbial ecosystems of the Precambrian era. We studied the environmental parameters of the boundary zone and examined the distribution of these types of mats and measured the in situ growth rates of the microorganisms composing them. In situ examination revealed that temperature and Eh constrain the development of the microbial mats. At the boundary between ST and CY, temperature and Eh ranged between 51.1°C and 63.2°C and between ?112 mV and ?25 mV, respectively. These environmental parameters were not significantly different among Japanese, Yellowstone (North American), and Icelandic hot spring effluents with genetically similar thermal sulfur oxidizers. Sickle-shaped bacteria rarely coexist with cyanobacteria, although they can potentially grow in some CY environments. This suggests that the boundary between ST and CY might be partly determined by exclusive ecological competition.     

Genomic and Functional Diversity of Bacteria Isolated from Hot Springs in Odisha,India

This study focused on culture-dependent survey of important bacterial community diversity of hot springs of Odisha. India. Molecular and cultural techniques were employed for assessing and exploiting the genetic and functional variability among the isolates obtained from three alkaline and mesophilic hot springs. A total of 48 isolates belonging to family Bacillaceae, Paenibacillaceae, Planococcaceae, Pseudomonadaceae and Enterobacteriaceae were identified. Majority of the bacterial isolates were affiliated with the genus Bacillus. Morphologically all the isolated bacteria were either Gram-positive spore-forming rods, or Gram-negative rods. The optimum temperature for growth of the isolates varied between 37°C to 50°C. The functional diversity revealed that many of the predominant and scarce isolates produced a variety of extracellular enzymes such as amylase, cellulase, lipase, phosphatase and protease, and genus Bacillus dominated for extracellular enzymatic activity. We employed two molecular markers to characterize the isolates. The hsp60 universal target sequence was found to be more discriminatory than 16s rRNA gene sequences. The cultivable bacterial community structure that colonized in the investigated thermal springs did not reveal much overlapping. Our results indicate that bacteria in the geothermal environment are metabolically active and cultivable populations may have great potential in biotechnology.     

The Effect of Temperature and pH on the Growth of Aerobic Alkalithermophilic Bacteria from Hot Springs in Buryatia

Growth parameters (temperature and pH) were determined for collection cultures of aerobic heterotrophic bacteria. Analysis of the experimental data with the use of the Rosso model made it possible to calculate the extreme values of temperature and pH permissive for culture growth. The cultures examined were subdivided into three groups with respect to their growth temperature and pH. The first group is represented by cultures with minimum, maximum, and optimal growth temperatures of <20, 60–64, and 38–40°C, respectively, and with the optimal growth pH 8.0–8.5. Bacteria of the second group are true alkalithermophilic organisms with a temperature optimum of 45–50°C and a pH optimum of 8.5–9.0. The third group includes a culture of a thermophilic alkalitolerant bacterium.     

白头鹞在中国的分类和分布讨论

白头鹞(Circus aeruginosus)和白腹鹞(C. spilonotus)的分类地位在过去曾存有争议,而分类上的变化有可能误导相关的记录。本文就白头鹞和白腹鹞的分类沿革进行了简要梳理,指出白腹鹞幼鸟及日本型与白头鹞的形态较为相近,并对白头鹞在国内的已知记录作了详细考证。结果发现,国内关于这两种鹞的历史记录确实因分类变化、辨识错误或文献解读而存在混淆。经仔细核查文献,辅以检视标本,认为白头鹞于新疆、北京、云南、贵州、青海、内蒙古和四川的记录存在有说服力的确实证据,其余地区的已知记录则应谨慎对待。     

华南地区龙眼的温度适宜性及其变化趋势

运用温度适宜度模型对华南地区龙眼生产和生育期的适宜性及其时空差异进行了计算、评价和预测。首先分析龙眼各生育期适宜度,结果表明,休眠期、花芽生理分化期、花芽形态分化期适宜性较低,具有较高的变率,开花期、果实发育成熟期、抽梢期表现出较高的温度适宜性,变率也较小,并且通过进一步研究发现影响华南地区龙眼生产的关键问题不是冷害,而是冬春季节的热害;其次分析华南地区各县区的平均温度适宜度,结果表明,华南地区龙眼温度适宜性的地域分异呈纬度地带性规律,并根据龙眼减产率将华南地区划分为最适宜型、适宜型、次适宜型、低适宜型和不适宜型五大适宜度类型;最后分析龙眼各生育期温度适宜度的年际变化,发现未来各生育期中除果实发育成熟期和抽梢期无明显变动外,其他生育期的温度适宜度都有下降趋势,同时对华南地区各站点龙眼温度适宜度的变化趋势作分析,结果表明,除福建省部分地区有弱上升外,总体上华南地区温度适宜度有下降趋势,且存在着地区差异,并且未来龙眼的适宜种植区将有可能北移,根据各站点未来适宜度变化的方向和强度进行分类,将华南地区划分为强下降型、次强下降型、弱下降型和上升型四大变动类型。     

Existence of Novel Phylotypes of Nitrite-Dependent Anaerobic Methane-Oxidizing Bacteria in Surface and Subsurface Sediments of the South China Sea

Nitrite-dependent anaerobic methane oxidation (n-damo) is a recently discovered new microbial process performed by the Candidatus Methylomirabilis oxyfera with an unusual intra-aerobic pathway, but there is no report about n-damo bacteria in marine environments. M. oxyfera-like sequences were successfully retrieved for the first time from both surface and subsurface ocean sediments of the South China Sea (SCS) using both 16S rRNA and pmoA genes as biomarkers and PCR amplification in this study. The majority of M. oxyfera-like 16S rRNA gene-based PCR amplified sequences from the SCS sediments formed a new group distinctively different from those detected in freshwater habitats and the information is consistent phylogenetically with those obtained from the pmoA gene. This study showed the existence of n-damo in ocean sediments and suggests that marine sediments harbor n-damo phylotypes different from those in the freshwater. This finding here expands our understanding on the distribution of n-damo bacteria to marine ecosystem and implies their potential contribution to the marine C and N cycling.     

A Window to the Subsurface: Microbial Diversity in Hot Springs of a Sulfidic Cave (Kaklik,Turkey)

Caves are windows to the extreme habitats of deep subsurface, and provide answers of unknowns about the underground life. Furthermore, sulfidic caves are important analogues for the early Earth environments, since some environmental conditions are common, such as high sulfur concentration, high temperature and oxygen-poor conditions. Kaklik Cave (Denizli, Turkey) with its travertine formation, carbonate- and sulfur-rich thermal springs, exhibits a unique ecosystem as a sulfidic cave. This study represents the first molecular survey of the microbial community in the Kaklik Cave, Turkey using high-throughput 16S rRNA gene amplicon sequencing analysis. An average of 859–2,416 operational taxonomic units per sample were observed including 25 bacterial phyla and 3 archaeal phyla. The bacterial diversity profiles were generally dominated by Epsilonproteobacteria and Gammaproteobacteria. At the carbonate-rich hot spring, that formed travertine structure, 9.7% of sequence reads affiliated with Thiofaba spp. In contrast, 38.74% of the total sequence reads at the sulfidic hot spring samples associated with the genus Sulfurimonas and Sulfurovum. In the archaeal community composition, Thermoplasmata was the most abundant group in all sampling areas. The 454-pyrotag results provide leads about ammonia-, nitrite- and sulfur-oxidation as well as sulfur-reduction, carbon dioxide fixation, and nitrogen fixation.     

西藏热泉沉积物的硫氧化细菌多样性及其影响因素

【目的】探究西藏热泉沉积物中硫氧化细菌群落多样性及其环境影响因素。【方法】选取西藏5个地热区的25个热泉样点,现场测量各采样点的水体理化参数,并采集热泉水和沉积物样品。通过功能基因(硫代硫酸盐水解酶编码基因:soxB)克隆文库分析方法,研究沉积物样品中硫氧化细菌群落的组成和多样性,并统计分析其与环境参数之间的相关性。【结果】克隆文库分析结果显示,西藏热泉沉积物样品中的硫氧化细菌主要分属于α-Proteobacteria、β-Proteobacteria和γ-Proteobacteria纲。研究样品中的硫氧化菌群落香农指数与溶解有机碳的浓度(R=0.489,P0.05)呈显著正相关。另外,统计发现西藏各热泉之间的硫氧化菌群落组成差异较大,其优势硫氧化细菌种群差异明显,大多数样点的优势硫氧化细菌种群为β-Proteobacteria,少量样点以α-Proteobacteria和γ-Proteobacteria为主。Mantel检验结果显示,西藏热泉样品中的硫氧化细菌群落组成与温度、硫化物、电导率、海拔、总溶解固体和pH显著(P0.05)相关。【结论】西藏热泉沉积物中的硫氧化细菌群落广泛分布,且主要以变形菌门为主。地理隔离和理化差异导致了热泉沉积物样品间的硫氧化菌群落组成差异。     

中国西部电网电击猛禽致死事故调查

2004～2008年调查了西部电网电击猛禽情况.调查区域包括新疆、青海、四川、甘肃、西藏等省区.共统计10 kV和35 kV输电线路长度178 km、杆塔数1 800余根;收集线路附近鸟的尸体、羽毛、残骸等110件,用于鉴定鸟种.调查结果,被电死猛禽计79只(10 kv),平均每千米输电线路1.36只,11.31%的杆塔对鸟类构成威胁;35 kV输电线路上鸟巢数44个,平均每千米输电线路O.37个,至少6.06%杆塔被猛禽利用.已知24%～40%的跳闸停电事故是由于鸟类引发的.症结分析,西部电网结构设计不合理.线路裸露,缺乏防鸟措施.在西部约20%左右的猛禽(11种)受到电击的威胁,同时严重影响着输电线路安全运行.本文提出了合理化建议.     

西南干热河谷地区葡萄休眠规律的研究

在西南干热河谷地区南亚热带气候条件下,4个酿酒葡萄品种梅尔诺 Merlot 、黑虎香 Fox 、霞多丽Chardonnay 和佳美 Gamay 分别在10月初、10月中旬、11月初和11月中旬进入生理性内休眠;约在12月初、12月中旬、1月初和12月中旬进入生态休眠阶段 萌芽率>5 0 % .休眠期葡萄新梢淀粉和总糖含量的变化趋势为:随着休眠程度的加深,淀粉含量上升,总糖含量下降;随着内休眠解除,总糖上升,淀粉下降,二者呈相反的变化趋势.研究结果为在该地区打破葡萄休眠,实现葡萄避雨栽培提供了理论基础.