Potential of Bioaugmentation and Biostimulation for Enhancing Intrinsic Biodegradation in Oil Hydrocarbon-Contaminated Soil

Studies were conducted using a 10-chamber Micro-Oxymax (Columbus, OH, USA) respirometer to determine the effect of bioaugmentation and biostimulation (by diverse ways of O₂ supply) on enhancing biodegradation of oil hydrocarbons to reduce risk at a former military airport in Kluczewo, Poland. Indigenous or exogenous bacteria bioaugmentation was used to degrade hydrocarbons. Aerated water and/or aqueous solutions of H₂O₂ or KMnO₄ were used to supply O₂. The intrinsic and enhanced biodegradation was evaluated by the O₂ uptake and CO₂ production rates obtained using a linear regression of the cumulative O₂ uptake and CO₂ production curves. Generally, in all cases biodegradation rates enhanced by bioaugmentation were two to four times higher than the rates of intrinsic biodegradation. Moreover, application of indigenous bacteria was more efficient in comparison to the exogenous consortia. The highest CO₂ production rates were achieved when aqueous solution of KMnO₄ was applied, as the increase of CO₂ production rates were about 71% to 97% higher compared to a control. The aqueous solution of H₂O₂ did not cause any significant improvement of the biodegradation rates. Compared to a control, the addition of aerated water resulted in a decrease of CO₂ production rates. Most probably the excessive soil moisture could reduce the air-filled porosity and, consequently, the oxygen contents in soil.     

Benzo[a]pyrene removal by Marasmiellus troyanus in soil microcosms

Benzo[a]pyrene (B[a]P) is a carcinogenic polyaromatic hydrocarbon that enters the environment as an incomplete combustion production of fossil fuels. Several species of filamentous fungi are capable of biotransforming and/or mineralizing B[a]P in liquid cultures, however there has been less success in soil habitats. In this study, the litter rot fungus Marasmiellus troyanus was encapsulated in alginate and delivered to B[a]P-spiked soil microcosms (100 μg B[a]P/g soil) for 1, 2 and 6 weeks, with and without a fertilizer solution. After 2 weeks, 32.5% of B[a]P was recovered from soil microcosms treated with M. troyanus compared to 55–70% for controls. After 6 weeks, controls demonstrated an average percent recovery of B[a]P of 54% while M. troyanus-inoculated samples gave an average percent recovery of 11%. Similar bioaugmentation of contaminated habitats with appropriately formulated fungi has potential for practical bioremediation in soil environments. Journal of Industrial Microbiology & Biotechnology (2000) 25, 116–119.     

闽江河口互花米草不同入侵阶段湿地土壤nirK型反硝化微生物群落结构及多样性

为了明确闽江河口互花米草海向不同入侵阶段湿地土壤nirK型反硝化微生物的群落结构及多样性,在鳝鱼滩东部的互花米草分布区,由陆向海方向选择互花米草海向入侵前的光滩(MF)、入侵1～2年(SAN)和入侵6～7年的互花米草(SA)湿地为研究对象,基于高通量测序技术,测定并分析了不同互花米草入侵阶段湿地土壤nirK型反硝化微生物群落结构及多样性的差异。结果表明:互花米草海向入侵降低了土壤中nirK型反硝化微生物群落的多样性及丰富度。不同入侵阶段湿地土壤的nirK型反硝化微生物主要包括变形菌门和放线菌门,其中变形菌门占绝对优势地位。互花米草海向入侵整体改变了湿地土壤中nirK型反硝化菌属的组成特征,MF、SAN、SA湿地土壤中相对丰度最高的nirK基因菌属分别为慢生根瘤菌属、中慢生根瘤菌属和产碱杆菌属。互花米草海向入侵增加了土壤中nirK型反硝化微生物群落组成的空间异质性,尤其是在SAN样地,这主要与样地本身的环境扰动性较大及互花米草海向入侵加大了样地环境因子的空间异质性有关。互花米草海向入侵主要是通过显著改变土壤理化因子(粒度组成、pH值和含水量)和氮养分条件(全氮、NH₄⁺-N、NO₃^--N)来影响nirK型反硝化微生物的群落结构及多样性。本研究结果有助于揭示互花米草海向入侵对湿地土壤反硝化过程影响的微生物机制。     

Investigation of Microbes in the Rhizosphere of Selected Trees for the Rhizoremediation of Hydrocarbon-Contaminated Soils

Hydrocarbon-degrading microorganisms (HDMs), associated with the rhizosphere of Conocarpus lancifolius and Ficus infectoria trees grown in bioremediated soil, were isolated under controlled laboratory conditions. The selected trees were used to phytoremediate oil-contaminated soil for three successive growing seasons. At the end of the phytoremediation experiment, 85.7% of measurable total petroleum hydrocarbon was degraded in the rhizosphere soil associated with Conocarpus lancifolius compared to 78.6% in the rhizosphere of Ficus infectoria. The detectable concentrations of some polyaromatic hydrocarbons were less than 0.02 ppm. The HDM isolation process was conducted at 35°C under aerobic conditions. The isolated HDMs were identified using 16S rRNA gene sequencing and fatty acid methyl ester analysis. Differences in the genera of the isolated HDMs and their assessed efficiency in degrading a mixture of hydrocarbon compounds between the two trees were noted.     

Bacterial and Archaeal 16S rRNA Genes in Late Pleistocene to Holocene Muddy Sediments from the Kanto Plain of Japan

Microbial communities in ancient marine sediments composed of clay and silt obtained from the terrestrial subsurface were phylogenetically analyzed based on their 16S rRNA gene sequences. Chloroflexi and Miscellaneous Crenarchaeotic Group were predominant in bacterial and archaeal clone libraries, respectively. Of 44 operational taxonomic units (OTUs) that had close relatives in the database, 30 were close to sequences obtained from marine environments. Some sequences belonged to the candidate groups JS1, ANME-I, and Marine Benthic Group-C, which are typically found in marine sediments. Low chloride concentrations in the sediments suggest that these marine-affiliated sequences may not reflect currently active microbial communities. Our results indicate the existence of long-term preserved DNA or descendants of ancient oceanic microbial components in subsurface muddy sediments in a temperate region, which may reflect indigenous population of paleoenvironments.     

广西北海西村港互花米草对红树林湿地大型底栖动物群落的影响

为了解互花米草(Spartina alterniflora)入侵红树林的生态影响, 作者对位于北海市西村港的红树林湿地以及周边互花米草盐沼的大型底栖动物群落多样性和群落结构进行了研究。2012年10月至2013年9月连续4次取样, 按照取样时间研究大型底栖动物的种类、物种组成、生物量和生物多样性等群落特征的差异, 探讨互花米草入侵红树林湿地对大型底栖动物的影响。本研究共采集底栖动物16种, 隶属于5门7纲15科, 其中互花米草群落10种, 红树林湿地12种。研究发现互花米草入侵后中国绿螂(Glauconome chinensis)个体数量剧增, 导致不同采样时间互花米草盐沼的大型底栖动物生物量均显著高于红树林湿地; 除个别月份外, 红树林湿地大型底栖动物的Margalef丰富度指数、Shannon-Wiener多样性指数、Simpson多样性指数和Pielou均匀度指数均显著高于互花米草群落。基于生境-采样时间的双因素方差分析结果表明, Shannon-Wiener多样性指数和Simpson指数在两种生境间差异显著; 两种生境的Margalef丰富度和Pielou均匀度指数在不同采样时间差异显著; 大型底栖动物生物量和物种数量在两种生境间和不同采样时间差异均显著。基于多元回归分析的研究结果表明, 互花米草密度是影响大型底栖动物生物量的关键因子, 而互花米草株高可以解释物种个体数量、Shannon-Wiener多样性指数和Simpson指数在两种生境的变化。对不同采样时间大型底栖动物群落结构的非度量多维度(non-metric multidimensional scaling, NMDS)分析结果表明, 红树林与互花米草群落的大型底栖动物群落相似性很低。总而言之, 在西村港地区, 互花米草入侵虽然增加了大型底栖动物的生物量, 但由于优势物种的凸显, 显著降低了大型底栖动物群落的多样性, 且种类组成与群落结构与红树林群落相比已有差异。由此可见, 互花米草入侵红树林对当地的大型底栖动物群落多样性造成影响。     

互花米草入侵九段沙河口湿地对当地昆虫多样性的影响

为认识因互花米草(Spartinaalterniflora)入侵而给九段沙河口湿地昆虫多样性带来的影响,我们于2004年5月到2005年10月间用网捕和收割植株两种方法对3个典型植物群落中昆虫多样性作了连续调查。研究期间,共采集到昆虫11,300头。经鉴定为97个种,隶属于12目69科。互花米草群落中昆虫的物种数、个体数量以及Shannon-Wiener多样性指数均显著低于土著植物芦苇(Phragmitesaustralis)群落和海三棱藨草(Scirpusmariqueter)群落中的;而Simpson优势度指数较土著植物群落中高。聚类分析结果表明:芦苇与海三棱藨草群落中昆虫群落结构更为相似。互花米草的入侵将可能导致九段沙湿地昆虫多样性的降低和群落结构的改变。     

Impacts of Bioremediation Schemes on Bacterial Population in Naphthalene-Contaminated Marine Sediments

Microcosm experiments were conduced in which the surface of marine sediment was contaminated with naphthalene and subjected to either of three different bioremediation schemes, i.e., biostimulation (BS) by supplementing with slow-release nitrogen and phosphorus fertilizers, bioaugmentation (BA) by inoculating with Cycloclasticus sp. E2, an aromatics-degrading bacterium identified to play an important role for aromatic-hydrocarbon degradation in marine environments and combination (CB) of BS and BA. These three schemes were found to be similarly effective for removing naphthalene, while naphthalene disappearance in sediment without any treatment (WT) was slower than those in the treated sediments. Shifts in bacterial populations during and after bioremediation were analyzed by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments. It was found that the Cycloclasticus rRNA type occurred as the strongest bands in the course of naphthalene degradation. Clustering analysis of DGGE profiles showed that bacterial populations in the WT, BS and CB sediments differed consistently from those in the uncontaminated control, while the profile for the BA sediment was finally included in the cluster for uncontaminated control sediments after a 150-day treatment. The results suggest that bioaugmentation with ecologically competent pollutant-degrading bacteria is an ecologically promising bioremediation scheme.     

Biodegradation of Pyrene in Marine Beach Sediments

The potential of chitosan (0.1% dry weight equivalent) as a bioremediation additive for removal of the recalcitrant polycyclic aromatic hydrocarbon (PAH) pyrene in marine beach sediments was investigated using an open irrigation system over a 63-day period. Osmocote, a slow release fertilizer, was used as the key nutrient supplement at a concentration of 1% in sediment (dry weight equivalent). Osmocote significantly (p < .05) enhanced nutrient levels, and the metabolic activity of the indigenous microbial biomass. Both additives were comparable in stimulating pyrene biodegradation rates; with chitosan (0.062 day^?1) being slightly more effective as an amendment than Osmocote (0.051 day^?1). Loss of pyrene in a control sediment (i.e., pyrene, without additives) was 66.6% over a 63-day period. The concurrent application of additives yielded the greatest biodegradation rates (0.072day^?1), resulting in a 98.2% loss of pyrene over 63 days. The treatment of oil contaminated beach sediments with both osmocote (1%) and chitosan (0.1%) is therefore recommended as an effective treatment for the intrinsic biodegradation of recalcitrant PAHs in oil-contaminated beach sediments.     

外来种互花米草对盐沼土壤微生物多样性的影响——以江苏滨海为例

研究了江苏滨海外来种互花米草的生长对潮间带土壤微生物特征的影响,结果显示:与原有光滩相比,互花米草的大面积生长,使当地潮间带土壤微生物量增加,并随植被的生长状况发生变化,潮间带土壤微生物对碳和营养物质的利用相对较少;同时微生物生理功能群中占优势的活动组分发生了变化,组成可能更复杂。土壤微生物群落生理功能多样性(CLPP)分析结果显示,潮间带土壤微生物群落对碳源底物的利用能力不高,所利用碳源以糖类、氨基酸类、羧酸类占优势;其中互花米草盐沼微生物利用的碳源种类多于光滩,并且不同季节土壤微生物群落碳源利用类型存在差异;潮间带土壤微生物在不同的季节中其生理功能群组成发生着变化,互花米草盐沼的土壤微生物在植被生长减弱时具有与光滩相近的代谢类群,在生理功能上具有一定的相似性;当植被生长旺盛时,其代谢类群增多,代谢强度升高,微生物群落功能多样性增加。外来种互花米草在滨海潮间带的大面积生长,改善了土壤理化性质,为该处土壤微生物提供了不同的碳源,增强了土壤微生物的活动,改变了土壤微生物群落生理功能结构。     

苏北大丰生态工程区两种植物群落土壤酶活性比较

土壤酶在土壤生态系统的物质循环和能量流动方面扮演着重要的角色.研究了大丰海滨湿地中实施微地貌水文饰变生态工程后的芦苇群落和互花米草群落土壤磷酸酶、脲酶和过氧化氢酶的季节变化动态,并对两种群落的3种土壤酶活性与各理化因子之间作了相关性分析,结果表明:(1)芦苇群落和互花米草群落土壤中磷酸酶活性季节变化动态均为单峰型曲线,过氧化氢酶活性均为单谷型曲线,最大值和最小值都出现在夏季(8月份),土壤脲酶活性季节变化规律不明显.(2)相关分析表明,互花米草群落土壤磷酸酶活性与土壤含盐量、植物生物量显著相关,而其过氧化氢酶活性与二者呈显著负相关;芦苇群落土壤磷酸酶活性与有机质和速效磷含量呈显著正相关,过氧化氢酶活性则与二者均呈显著负相关.此外,两种群落中的土壤脲酶活性与所测土壤理化因子均无明显相关关系.(3)两种植物群落中土壤酶活性受土壤理化因子的共同影响.通径分析表明,在互花米草群落中,各因子对磷酸酶活性的影响程度依次为:土壤含盐量＞速效钾＞速效磷＞有机质＞铵态氮;对脲酶活性的影响程度依次为:有机质＞速效钾＞速效磷＞土壤含盐量＞铵态氮;对过氧化氢酶活性的影响程度为:速效钾＞土壤含盐量＞有机质＞速效磷＞铵态氮.在芦苇群落中,各因子对磷酸酶活性的影响程度依次为:速效磷＞土壤含盐量＞有机质＞铵态氮＞速效钾;对脲酶活性的影响程度依次为:有机质＞速效钾＞土壤含盐量＞铵态氮＞速效磷;对过氧化氢酶活性的影响程度为:铵态氮＞速效钾＞土壤含盐量＞速效磷＞有机质.(4)微地貌水文饰变生态工程在一定程度上改良了土壤,有利于实现芦苇对互花米草的替代.     

Bacterial Diversity among the Sediments of Glacial Lakes in the Western Balkans: Exploring the Impact of Human Population

16S rRNA gene-based metagenomic approach was used to assess the biodiversity of bacterial communities in the sediments of selected glacial lakes in the Western Balkans and to assess the impact of human population on these microbial communities. Sediment samples were collected from three glacial lakes, viz., Plav Lake (in a zone of the highest impact of human population), Black Lake (a zone of medium impact of human population), and Donje Bare Lake (a remote lake with minimal impact of human population).

Canonical correlation analysis analysis indicated correlation between the distance of the lake from urbanized population and bacterial diversity in Donje Bare Lake sediment. Bacterial diversity of Black Lake sediment was correlated with high content of phosphorous and pH value. Chemical compounds exhibiting the most prominent correlation with bacterial diversity of Plav Lake were NH₄-N, K₂O, CaCo₃, and total nitrogen . Additionally, CCA analysis indicated that population density was correlated with biodiversity of bacterial communities in Plav Lake sediment, which is the most exposed to human population. Multivariate regression revealed the highest correlation between the presence of Proteobacteria classes and population density and levels of NH₄-N.

The influence of human population was observed to be important for shaping the sediment communities in addition to biological and chemical factors.     

互花米草入侵对黄河口潮沟形态特征和植物群落分布的影响

互花米草大规模入侵滨海湿地,对潮沟形态特征和植物群落的演变产生重大影响.本研究基于2008-2020年遥感影像,利用3S技术,结合回归分析与统计分析法,分析黄河口湿地互花米草入侵的演变特征,探究其对潮沟形态特征和植物群落分布的影响.结果 表明:2008年以来,黄河口互花米草面积增长迅速,增长速度为4.47 km2·a-...     

Bacterial Diversity and Geochemical Profiles in Sediments from Eutrophic Azorean Lakes

In the Azores, the advanced trophic state of the lakes requires a fast intervention to achieve the good ecological status prescribed by the Water Framework Directive. Despite the considerable effort made to describe the phytoplankton growing on the water column, the lack of information regarding the microbial processes in sediments is still high. Thus, for the successful implementation of internal management actions, the present work explored the relationships between geochemical profiles and dominant members of the bacterial community in sediments from eutrophic Azorean lakes. Lake Azul geochemical profiles were quite homogeneous for all parameters, while in lake Furnas the total iron profile presented a peak below the aerobic layer. For lake Verde, the concentrations of all studied parameters (20 ± 2% loss-on-ignition; 2.10 ± 0.08 mg g^?1 total phosphorus; 1.31 ± 0.50 mg g^?1 total nitrogen; 8.06 ± 0.13 mg g^?1 total iron) in the uppermost sediment layer were approximately two times higher than the ones in sediments from other lakes, decreasing with sediment depth. The higher amounts of phosphorus and organic matter in lake Verde suggested a higher internal contribution of phosphorus to eutrophication. The dominant members of the sediment bacterial community, investigated by denaturing gradient gel electrophoresis, were mostly affiliated to Proteobacteria phylum (Alpha-, Delta-, and Gamma-subclasses), group Bacteroidetes/Chlorobi and phylum Chloroflexi. The Cyanobacteria phylum was solely detected in sediments from lake Verde and lake Furnas that presented the highest amounts of nitrogen and phosphorus both in the water column and sediments, while the other phyla were detected in sediments from the three studied lakes. In conclusion, management measurers to achieve the good ecological status until 2015 should be distinct for the different lakes taking into account the relative magnitude of the nutrient sources and the bacterial diversity in sediments.     

Vertical distribution of nitrite reductase genes (nir S) in continental margin sediments of the Gulf of Mexico

Marine sediments account for up to 66% of the loss of nitrogen load to coastal areas. Sedimentary denitrification is the main sink for fixed nitrogen in the global nitrogen budget, and thus it is important to understand the structure and composition of denitrifying communities. To understand the structure and composition of denitrifying communities, the diversity of nitrite reductase (nirS) genes from sediments along the Gulf of Mexico was examined using a PCR-based cloning approach. Sediments were collected at three different depths (0-0.5, 4-5 and 19-21 cm). Geochemical analysis revealed decreasing nitrate and oxygen concentrations with increasing sediment depth. This trend coincided with the decrease in diversity of denitrifying bacteria. LIBSHUFF analysis indicated that the clone library in the shallowest sediment (depth, 0-0.5 cm) was significantly different from that in the deepest sediment (depth, 19-21 cm), and that the deeper sediments (depths of 4-5 and 19-21 cm) were significantly similar. Community structural shifts were evident between the shallowest (oxic zone) and deepest (anoxic zone) sediments. Community changes within the deepest sediments were more subtle, with the presence of different nirS clone sequences gradually becoming dominant or, alternatively, decreasing with depth. The changes in community structure at this depth are possibly driven by nutrient availability, with lower quality sources of carbon and energy leading to the disappearance of nirS sequences common in the top layer. The majority of recovered nirS sequences were phylogenetically divergent relative to known denitrifying bacteria in the database.     

Bioremediation of Diesel Fuel Contaminated Soil: Comparison of Individual Compounds to Complex Mixtures

Bioremediation is a cost effective means of remediating soils contaminated with petroleum hydrocarbons. Various factors affect the efficiency of the process, including environmental conditions, microbial population present and composition of the hydrocarbon spill. To evaluate the impact of mixture composition on individual compound and overall degradation, biodegradation experiments were conducted in sealed, 1-liter bioreactor/respirometer vessels containing soil spiked with hydrocarbon compounds in isolation and in mixtures. The influence of bacteria and fungi on the degradation process was also monitored. The degradation behavior of the various compounds was monitored using the fraction of contaminant remaining and first-order degradation coefficients based on hydrocarbon loss.

The results showed that the degradation trend of a compound changed when present in a simple mixture or when degraded in isolation. The presence of the compounds as either an aliphatic or aromatic mixture did not change the degradation trend. The presence of a mixture also affected the amount of degradation with some compounds degrading to a greater extent when in isolation. Overall, the majority of degradation occurred in the first 10 d, suggesting that the first-order model may not be an appropriate model for degradation periods longer than 10 d when nutrient limited. It was also found that fungal metabolism is important for the degradation of hydrocarbons, particularly for branched species such as pristane as the decay rate increased one order of magnitude when bacteria were inhibited.     

Microbial Communities of Deep Marine Subsurface Sediments: Molecular and Cultivation Surveys

Recent molecular analyses show that microbial communities of deep marine sediments harbor members of distinct, uncultured bacterial and archaeal lineages, in addition to Gram-positive bacteria and Proteobacteria that are detected by cultivation surveys. Several of these subsurface lineages show cosmopolitan occurrence patterns; they can be found in cold marine sediments and also in hydrothermal habitats, suggesting a continuous deep subsurface and hydrothermal biosphere with shared microbiota. The physiologies and activities of these uncultured subsurface lineages remain to be explored by innovative combinations of genomic and biogeochemical approaches.     

多环芳烃污染土壤微生物修复研究进展

多环芳烃是我国土壤环境质量标准中要求严格管控的一类持久性有机污染物,利用微生物技术修复有机污染土壤具有绿色、经济等突出特点,应用前景广泛。目前多学科的协同发展和新技术的研究应用,为多环芳烃土壤微生物转化机制与污染生态过程等方面带来了新的认识,同时对修复技术的实际应用与调控提供了新的思考方向。本文以多环芳烃污染土壤微生物修复为主体,从污染土壤微生物修复应用技术、多环芳烃微生物降解特征、土壤体系污染物归趋规律与微生物作用及土壤污染微生物群落响应与研究技术等方面进行综合评述,并针对现存应用技术瓶颈和理论空白作进一步思考和展望。     

Isolation and phylogenetic analysis of cultivable manganese bacteria in sediments from the Arctic Ocean

Isolation, molecular identification and phylogenetic analysis were carried out to investigate the biodiversity of manganese bacteria in sediments which were collected from the Arctic Ocean during the 2nd Chinese Arctic Scientific Expedition. 21 and 19 species of cultivable strains were isolated from sediments at Stations P11 and S11, respectively, according to their distinct morphological character on the screening plate of manganese medium. Molecular identification and phylogenetic analysis showed that the cultivable manganese bacteria from Station P11 were basically composed of γ-Proteobacteria (γ subgroup of the Proteobacteria branch of the domain Bacteria) and Actinobacteria, which accounted for 86% and 14%, respectively. The isolates of γ-Proteobacteria mainly included Psychrobacter, Shewanella, Acinetobacter and Marinobacter, of which Psychrobacter was the major genus, which accounted for 67% of the γ-Proteobacteria. The cultivable manganese bacteria from Station S11 included α-Proteobacteria, γ-Proteobacteria and Flavobacteria of Bacteroides. The γ-Proteobacteria mainly included Shewanella, Marinomonas and Alteromonas. The majority of α-Proteobacteria was Sphingomonas. The phylogenetic analysis indicated that bacteria from sediments at Stations P11 and S11 had different cultivable manganese microbial communities. All tested strains had higher resistance to Mn2+, of which Marinomonas sp. S11-S-4 had the highest resistant ability.