Fe- and S-Metabolizing Microbial Communities Dominate an AMD-Contaminated River Ecosystem and Play Important Roles in Fe and S Cycling

Indigenous Fe- and S-metabolizing bacteria play important roles both in the formation and the natural attenuation of acid mine drainage (AMD). Due to its low pH and Fe-S-rich waters, a river located in the Dabaoshan Mine area provides an ideal opportunity to study indigenous Fe- and S-metabolizing microbial communities and their roles in biogeochemical Fe and S cycling. In this work, water and sediment samples were collected from the river for physicochemical, mineralogical, and microbiological analyses. Illumina MiSeq sequencing indicated higher species richness in the sediment than in the water. Sequencing also found that Fe- and S-metabolizing bacteria were the dominant microorganisms in the heavily and moderately contaminated areas. Fe- and S-metabolizing bacteria found in the water were aerobes or facultative anaerobes, including Acidithiobacillus, Acidiphilium, Thiomonas, Gallionella, and Leptospirillum. Fe- and S-metabolizing bacteria found in the sediment belong to microaerobes, facultative anaerobes, or obligatory anaerobes, including Acidithiobacillus, Sulfobacillus, Thiomonas, Gallionella, Geobacter, Geothrix, and Clostridium. Among the dominant genera in the sediment, Geobacter and Geothrix were rarely detected in AMD-contaminated natural environments. Canonical correspondence analysis indicated that pH, S, and Fe concentration gradients were the most important factors in structuring the river microbial community. Moreover, a scheme explaining the biogeochemical Fe and S cycling is advanced in light of the Fe and S species distribution and the identified Fe- and S-metabolizing bacteria.     

A Novel Community of Acidophiles in an Acid Mine Drainage Sediment

A sediment sample (pH 2.5) was collected at an acid mine drainage site in Anhui, China. The present acidophilic microbial community in the sediment was studied with a 16S rRNA gene clone library. Small-subunit rRNA genes were PCR amplified, cloned and screened by amplified rDNA restriction analysis (ARDRA). Subsequently, 10 different clones were identified and they were affiliated with Acidobacteria, β/γ-Proteobacteria, δ-Proteobacteria, Nitrospira, Candidate division TM7, and Low G + C Gram-positives. Phylogenetic analysis of 16S rRNA gene sequences revealed a diversity of acidophiles in the sediment that were mostly novel. Unexpectedly, 16S rRNA gene sequences affiliated with δ-Proteobacteria were found to constitute more than 60% of clone library. To our knowledge, this is the first occasion that bacteria of δ-Proteobacteria have been found dominant in the acidic habitat. Anaerobic sulfate- or metal reduction is the predominant physiological trait of bacteria of this subdivision. The high sulfate, ferric iron and the presence of bioavailable carbon in the anaerobic microenvironment may result in the dominance of bacteria of δ-Proteobacteria.     

Culturable and molecular phylogenetic diversity of microorganisms in an open-dumped,extremely acidic Pb/Zn mine tailings

A combination of cultivation-based and molecular-based approaches was used to reveal the culturable and molecular diversity of the microbes inhabiting an open-dumped Pb/Zn mine tailings that was undergoing intensive acid generation (pH 1.9). Culturable bacteria found in the extremely acidic mine tailings were Acidithiobacillus ferrooxidans, Leptospirillum ferriphilum, Sulfobacillus thermotolerans and Acidiphilium cryptum, where the number of acidophilic heterotrophs was ten times higher than that of the iron- and sulfur-oxidizing bacteria. Cloning and phylogenetic analysis revealed that, in contrast to the adjacent AMD, the mine tailings possessed a low microbial diversity with archaeal sequence types dominating the 16S rRNA gene library. Of the 141 clones examined, 132 were represented by two sequence types phylogenetically affiliated with the iron-oxidizing archaea Ferroplasma acidiphilum and three belonged to two tentative groups within the Thermoplasma lineage so far represented by only a few environmental sequences. Six clones in the library were represented by the only bacterial sequence type and were closely related to the well-described iron-oxidizer L. ferriphilum. The significant differences in the prokaryotic community structures of the extremely acidic mine tailings and the AMD associated with it highlights the importance of studying the microbial communities that are more directly involved in the iron and sulfur cycles of mine tailings.     

封闭环境下酸性矿坑水中微生物生态多样性的研究

铜绿山铜矿是世界开采时间最长的矿井之一,在开采过程中有许多矿井被废弃,许多废弃的矿井内产生了大量的对环境有害的酸性矿坑水.酸性矿坑水取自铜绿山铜矿某废弃矿井,利用限制性酶切片断多样性分析(RFLP分析)对酸性矿坑水中的微生物生态多样性进行了研究.研究表明,酸性矿坑水呈酸性,相对于其他极端与非极端生态环境,酸性矿坑水中的细菌与古菌的群落多样性较低.RFLP分析与系统发育分析表明,酸性矿坑水中细菌主要由A.fcrrooxidans(属于gamma-Proteobacteria)和L.ferrooxidans(属于Nitospira)成;古菌主要由Thermoplasma相关古菌组成.在这种封闭环境的酸性矿坑水中首次发现了类似于产甲烷古菌的克隆片断,其占古菌种群的四分之一左右.本研究将促进对酸性矿坑水中细菌及古菌群落组成及其对酸性矿坑水产生的作用的研究.     

紫金山铜矿酸性矿山废水微生物群落多样性

[背景]为避免环境污染,酸性矿山废水需经处理后才能排放,处理后的废水理化性质会发生显著变化,将影响整个微生物群落的结构.[目的]分析处理前后的细菌和真菌群落变化及其与理化参数的关系,为矿山废水的处理提供参考指标,并为矿山污染场地的修复提供理论基础.[方法]采集福建紫金山铜矿的酸性矿山废水并测定其理化性质.采用基于原核微...     

安徽省某酸性矿山废水坑湖中细菌群落的垂向分布特征及其影响因素

【目的】探究酸性矿山废水(acid mine drainage,AMD)坑湖中细菌群落沿垂向不同水深的分布规律及与环境因子之间的相互作用。【方法】采用16SrRNA基因高通量测序技术,对安徽省某AMD坑湖中6条采样垂线不同水深深度的细菌群落进行调查,同时测定水质理化指标,使用统计学软件分析细菌和地化参数间的联系。【结果】AMD坑湖中水质特征及细菌群落结构出现明显分层现象,自上而下溶解氧降低而pH和多种金属离子浓度增加,微生物群落结构发生变化,多样性和部分物种的丰度增大。细菌群落组成上,表层水域以Proteobacteria (Alpha、Gammaproteobacteria)和Acidobacteria占据主导地位;中下层水域则由Firmicutes、Acidobacteria、Actinobacteria、Gammaproteobacteria和Patescibacteria等共同主导。统计分析结果表明,TN、DO、ORP、pH、Fe、Mn、Al和Zn与嗜酸细菌丰度显著相关,是细菌空间分布的主要限制因素。【结论】AMD坑湖中水质理化特征和细菌群落分布在垂向空间上存在显著差异,群落的...     

Macrofilamentous microbial communities in the metal-rich and acidic River Tinto, Spain

A novel type of macroscopic microbial community consisting of large dendritic filaments (up to 1.5 m) in a pH 2.0 dam of the River Tinto (South-western Spain) is described. The combined use of 16S rRNA-gene surveys and fluorescent in situ hybridisation (FISH) suggested that gamma-proteobacteria and a relative large diversity of alpha-proteobacteria dominated these structures. beta-Proteobacteria, Actinobacteria and Firmicutes were also detected. Whereas acidophilic bacteria of the genera Acidithiobacillus, Leptospirillum and Acidiphilium, and archaea belonging to the Thermoplasmatales dominate mine acid drainage waters and streamers (riverbed filamentous biofilms), none of the lineages identified in this study affiliate to typical acid mine drainage acidophilic bacteria. Bacteria of the Tinto macrofilaments might be heterotrophic, and could be feeding on the organic matter entrapped in the filamentous structure.     

纳米Fe3O4对生菜生长及土壤细菌群落结构的影响

纳米Fe₃O₄的广泛应用增加了其暴露到农田环境的可能性,因此亟待研究纳米Fe₃O₄对农业生态环境的影响.本研究采用盆栽试验方式,研究不同浓度纳米Fe₃O₄颗粒(1、10、100 mg·kg^-1)对生菜生长及土壤细菌群落的影响,并与相应浓度的普通Fe₃O₄处理进行对比.通过测定植物光合速率常数、植株Fe含量来评价植物生长;采用高通量测序技术研究土壤细菌群落结构及组成.结果表明: 不同浓度纳米Fe₃O₄的影响不同.低浓度纳米Fe₃O₄能提高植物生物量,增强植物叶片光合速率,增加土壤中黄单胞菌目的相对丰度,降低蓝细菌、鞘脂杆菌纲的相对丰度,但对群落多样性指数影响不显著.高浓度纳米Fe₃O₄抑制作物生长,提高植株中Fe积累及土壤电导率,降低细菌群落系统发育多样性,降低黄单胞菌目、鞘脂杆菌纲相对丰度,增加蓝细菌相对丰度.此外,一些土壤功能微生物对纳米Fe₃O₄及普通Fe₃O₄处理的响应也存在差异,说明不同粒径及浓度的Fe₃O₄均会对土壤微生物群落产生影响,并可能影响地上部分植物性状.因此,在评估纳米颗粒的生物学效应时需较多关注土壤微生物.     

Short- versus Long-Term Effects of Elevated CO2 on Night-Time Respiration of Needles of Scots Pine (Pinus sylvestris L.)

Dark respiration rate in the night (R _D) was measured in five-year-old Scots pine (Pinus sylvestris L.) trees grown for two years under ambient (AC) and elevated (AC + 400 µmol mol^–1 = EC) CO₂ concentrations in open top chambers. Two needle age classes (i.e., current-year and one-year-old) were measured at AC and EC in both AC- and EC-grown pines. Additionally different chemical characteristics were determined on the needles, such as nitrogen (N), carbon (C), starch, and soluble sugar concentrations as well as specific leaf area. The direct, short-term and indirect, long-term effects of EC on R _D for the two needle age classes were examined. R _D was expressed on a per needle area, needle mass, N, C, and C/N bases. Direct effects were only pronounced in the AC treatment where inhibition of R _D was found at EC in both current- and one-year-old needles. Indirect effects were only significant in one-year-old needles where a decrease was found in the EC grown trees as compared with AC ones when R _D was expressed per unit needle mass, C, or C/N. R _D per unit needle area and needle N were not sensitive to long-term EC, in any needle age class. Long-term EC treatment also influenced the response of the two needle age classes. One-year-old needles from the EC treatment had significantly lower R _D than current-year needles, but no such response was observed in the AC treatment. Our experiment re-emphasised the importance of expressing R _D on different bases for a correct interpretation of the responses to EC. Moreover, we showed that different needle age classes can respond differently to a CO₂ enrichment.     

Spatial comparison of total vs. active bacterial populations by coupling genetic fingerprinting and clone library analyses in the NW Mediterranean Sea

Spatial distributions of both total (i.e. 16S rDNA-based fingerprints) and active (i.e. 16S rRNA-based fingerprints) bacterial populations, together with total bacterial activity measured by ³H-leucine incorporation, were studied along a 98 km transect in the NW Mediterranean Sea. Capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) fingerprinting was coupled to a clone library, allowing CE-SSCP peaks identification and the monitoring of the spatial variation of bacterial phylotypes. Up to 80% of the community peaks matched those obtained from clone library sequences, accounting for 86.7% of the total fingerprinting area. A good agreement was found between the relative abundance of Prochlorococcus in the CE-SSCP fingerprints and flow cytometry counts ( r ²=0.66, P <0.05). The largest differences between total and active bacterial populations distribution were found at depths with higher bacterial activity (i.e. surface and deep chlorophyll maximum, DCM). SAR11 at the surface and Gammaproteobacteria at the DCM were the most abundant groups on the 16S rDNA-based fingerprints. However, their ratio of relative importance between rRNA : rDNA was <1 in most cases. Conversely, ratios observed for Prochlorococcus , were consistently >1 both at the surface and at the DCM. These results emphasize the need for combining rDNA- and rRNA-based analyses to better understand the functional role of individual bacterial populations in situ .     

外来入侵杂草的生物防治及生防因子对本地非目标种的影响

随着生物入侵所引起的生态及经济问题日益严重,对有害入侵生物的防治问题也备受人们关注。生物防治因具有持续、高效、安全等优点,已成为防治有害入侵生物的重要方法。传统生防是防治有害入侵杂草的一种重要方法。在简单介绍生物防治的基础上,重点阐述了传统生物防治的理论基础——天敌逃逸假说,生防因子对外来人侵种的影响及其对本地非目标种的直接和间接效应,并针对这些问题,对我国开展生物防治工作提出几点建议。     

Bacterial Diversity and Community Structure in High Arsenic Aquifers in Hetao Plain of Inner Mongolia,China

The bacterial diversity and community structure of high arsenic (As) aquifers was investigated using an integrated approach adopting both geochemistry and molecular biology (polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analyses). Nine borehole sediments and one groundwater sample from the living place of a villager (affected by arseniasis) and 12 sediments from a control borehole in Hetao Plain were investigated. The As concentrations ranged from 33.6 to 77.6 mg/kg in high As borehole sediments and 1.5 to 5.8 mg/kg in those samples from the control. The As concentration in the groundwater was 744.8 μg/L. Ratios between As(III) and total As in high As sediments increased gradually with depth and ranged from 0.02 to 0.34. Similarly, the Fe(II)/total Fe presented the same increasing trend with depth. The correlation between TOC contents and total As was positive. High concentrations of total As, S, Fe and TOC were found in clay and low in sand samples. Phylogenetic analysis showed significantly different bacterial communities among high As sediments, control sediments and the high As groundwater. Both DGGE and 16S rRNA gene clone library results showed that the high As sediments were dominated by Thiobacillus, Pseudomonas, Brevundimonas, and Hydrogenophaga, with Thiobacillus being distinctly dominant (63.5%). Whereas the low As sediments were dominated by some other genera including Psychrobacter, Massilia and Desulfotalea. The bacterial populations in the high As groundwater mainly included Pseudomonas, Acinetobacter and Aquabacterium. These results improve our understanding of the bacterial diversity in high As aquifers in Hetao Plain and suggest how specific bacterial populations help mediate the mobilization of As into high As groundwaters.     

16S rRNA gene analyses of bacterial community structures in the soils of evergreen broad-leaved forests in south-west China

Bacterial community structure was studied in humus and mineral soils of evergreen broad-leaved forests in Ailaoshan and Xishuangbanna, representing subtropical and tropical ecosystems, respectively, in south-west China using sequence analysis and terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Clone sequences affiliated to Acidobacteria were retrieved as the predominant bacterial phylum in both forest soils, followed by those affiliated to members of the Proteobacteria, Planctomycete and Verrucomicrobia. Despite higher floristic richness at the Xishuangbanna forest than at the Ailaoshan forest, soil at Xishuangbanna harbored a distinctly high relative abundance of Acidobacteria-affiliated sequences (80% of the total clones), which led to a lower overall bacterial diversity than at Ailaoshan. Bacterial communities in humus and mineral soils of the two forests appeared to be well differentiated, based on 16S rRNA gene phylogeny, and correlations were found between the bacterial T-RFLP community patterns and the organic carbon and nutrient contents of the soil samples. The data reveal that Acidobacteria dominate soil bacterial communities in the evergreen broad-leaved forests studied here and suggest that bacterial diversity may be influenced by soil carbon and nutrient levels, but is not related to floristic richness along the climatic gradient from subtropical to tropical forests in south-west China.     

Differential selection of growth rate-related traits in wild barley, Hordeum spontaneum, in contrasting greenhouse nutrient environments

Across-species comparisons show that inherent variation in relative growth rate (RGR) and its underlying traits are correlated with habitat productivity. In this study, we test the hypothesis that growth rate-related traits confer differential selective effects in contrasting nutrient environments. We specifically test whether high RGR is targeted by selection in nutrient-rich environments whereas low values of traits that underlie RGR [specific leaf area (SLA), leaf mass fraction and leaf area ratio (LAR)] confer a direct fitness advantage in nutrient-poor environments, resulting in selection of low RGR as a correlated response. We measured RGR, its underlying component traits, and estimated fitness in a range of wild barley (Hordeum spontaneum) accessions grown under high and low nutrient conditions. Selection on component traits differed between the two environments, while total selection of RGR was not significant. Using multiple regression and path analysis to estimate direct fitness effects, a selective advantage of high LAR and SLA was demonstrated only under nutrient-rich conditions. While supporting the view that observed associations between habitat richness and some RGR-component traits reflect adaptation to differing nutrient regimes, our data suggest that direct selection targets component traits rather than RGR itself.     

Estimating red deer abundance in a wide range of management situations in Mediterranean habitats

When human interventions interfere with the natural regulation of wildlife populations by favouring some species, overabundance can emerge. We evaluated different methods of estimating red deer abundance in a wide range of population densities from southern Spain. Distance sampling estimates were used as the reference method across 22 localities and were compared with two kilometric abundance indices (KAIs), four indices based on pellet group counts and two browsing indices (BWIs). The average red deer density estimated by distance sampling was 19.51±3.19 deer per 100 ha, showing a wide range across the study area (0.04–66.77). Distance sampling estimates correlated with the KAIs, pellet group-based index and the BWI. The agreement with distance sampling improved when groups were used instead of individuals in the KAIs, when the minimum pellet group size was fixed at 20 pellets in the dropping counts, and when only highly palatable species were used in the BWI. Thus, several direct and indirect methods can estimate red deer abundance in Mediterranean habitats from Southern Spain with appropriate modifications.