Studies on the solubilization of German coal by fungi

Summary The capability of seven basidiomycetes (Trametes versicolor, Poria placenta, Pleurotus florida, P. ostreatus, P. sajor-caju, P. eryngii, Stropharia sp.), one ascomycete (Chaetomium globosum) and five hyphomycetes and moulds (Humicola grisea, Trichoderma viride, Aspergillus terreus, Paecilomyces varioti, Papulaspora immersa) to solubilize medium and high volatile bituminous coals (types A and B) as well as four types of lignite B from Germany was tested in surface cultures. The intensity of bioliquefaction was determined by estimating the rate of droplet formation and by measuring the loss of weight of the coal granules gravimetrically. The bituminous coals with a relative high degree of coalification were only moderately converted by Trametes versicolor, Pleurotus florida, P. ostreatus and P. sajor-caju. The three species of Pleurotus caused the greatest rate of biosolubilization of lignite, yielding a loss of weight of the coal granules of more than 5.8% with a maximum of 7.6% with P. florida. The non-basidiomycetes proved to be less active with a liquefaction rate of up to 3.5% with Trichoderma viride. In general, the geologically younger lignite coals were more effectively solubilized than the older hard coals. The volatile matter and the oxygen content proved to be the principal factors influencing the intensity of bioconversion.     

Simulated Herbivory and Vegetation Dynamics in Coal Slurry Ponds Reclaimed as Wetlands

The biodiversity of coal slurry ponds can be inhibited, at least in part, by dense stands of Phragmites australis. In this study, we demonstrate that species richness can be increased in coal slurry ponds if the dominant species (P. australis and Typha latifolia) are removed and that underwater herbivory simulated by cutting will kill emergents. The study was conducted in the greenhouse and the field in both flooded and drawndown conditions. Stems of plants of P. australis and T. latifolia were cut in a greenhouse and the cut plants of both species showed a decline in survivorship (25 and 42% survival, respectively) whereas all uncut plants survived. In a reclaimed coal pond at Pyramid State Park, Illinois, neither P. australis nor T. latifolia survived cutting underwater, but all of the uncut plants survived. Regrowth measured as total biomass of stems was less among flooded versus freely drained plants (0.3 and 2.6 g biomass, respectively). Cut versus uncut plants, combining freely drained and flooded, had less below-ground biomass (99.4 and 254.4 g, respectively). In the greenhouse study, oxygen levels in rhizomes subsequent to cutting were measured using an oxygen electrode and millivolt meter. Oxygen levels in P. australis were lower in cut versus uncut plants both in flooded (15.0 vs. 16.3% ambient O₂, respectively) and freely drained conditions (14.5 vs. 15.0%, ambient O₂, respectively). Similar responses to cutting were demonstrated by T. latifolia. In an unreclaimed coal slurry pond with monospecific stands of P. australis, plant species richness increased in cut plots as compared to uncut plots (29 vs. 2 species, respectively) between March and September, 1995. This study demonstrated that species richness can be increased in coal ponds by mechanical cutting and this potentially by herbivory; however, the additional species were mostly exotics.     

Biodepolymerization studies of low rank Indian coals

Biodepolymerization of some of the lower rank Indian coals by Pleurotus djamor, Pleurotus citrinopileatus and Aspergillus species were studied in a batch system. The main disadvantage in burning low rank coals is the low calorific values. To get the maximum benefit from the low rank coals, the non fuel uses of coals needs to be explored. The liquefaction of coals is the preliminary processes for such approaches. The present study is undertaken specifically to investigate the optimization of bio depolymerization of neyveli lignite by P. djmor. The pH of the media reached a constant value of about 7.8 by microbial action. The effect of different carbon and nitrogen sources and influence of chelators and metal ions on depolymerization of lignite were also studied. Lignite was solubilized by P. djamor only to a limited extent without the addition of carbon and nitrogen sources. Sucrose was the best suitable carbon source for coal depolymerization by P. djamor and sodium nitrated followed by urea was the best nitrogen source. The Chelators like salicylic acid, TEA and metal ions Mg²⁺, Fe³⁺, Ca²⁺, Cu²⁺, Mn²⁺ has enhanced the lignite solubilization process. The finding of the study showed that, compared to sub-bituminous and bituminous coal, the lignite has higher rate of solubilization activity.     

Chemical reactivity of the carbon-centered free radicals and ferrous iron in coals: Role of bioavailable Fe2+ in coal workers' pneumoconiosis

Striking differences in the prevalence of coal workers' pneumoconiosis (CWP) exist between different coal mine regions. The major factors responsible for the observed regional differences in CWP have not yet been identified. In the present study, chemical reactivity of the carbon-centered free radicals in coals and lung tissues, as well as ferrous iron in the coals, were studied by ESR techniques. The ESR spectra clearly demonstrated the presence of at least two types of carbon-centered free radical species, which might respectively attribute to the macromolecular phase and the molecular phase of coal. Grinding produced free radicals in coals. Exposure of freshly ground coal to air for 28 h induced a slight increase of free radicals for most of the coals, and a slight decrease after 4 months' exposure. The lung tissue samples of coal workers deceased of CWP showed similar ESR spectra as coal samples, and these radicals were highly stable in the lung. After incubation of coals with glutathione, hydrogen peroxide, sodium formate or oxygen, the coal sample from the Gardanne mine which has never induced CWP, and thus is the least hazardous coal, showed the most significant change in the carbon-centered free radical concentration. No significant changes were observed among other coals reported to induce CWP. On the other hand, we found that the coals released different amounts of Fe²⁺ in an acidic medium. Interestingly, the prevalence of CWP correlates positively with the released Fe²⁺ content in these coals and with the amount of oxygen radicals produced by the interaction of Fe²⁺ with O₂ in the acidified coal filtrates. Our studies indicate that the carbon-centered free radicals may not be biologically relevant to coal dust-induced pneumoconiosis, whereas the acid soluble Fe²⁺, which may be dissolved in the phagolysosomes of macrophages, can then lead to Fe²⁺-induced oxidative stress and eventual CWP development.     

Colonization and degradation of oxidized bituminous and lignite coals by fungi

Summary APenicillium sp. previously shown to grow on lignite coals degraded an air-oxidized bituminous coal (Illinois #6) to a material that was more than 80% soluble in 0.5 N NaOH. Scanning electron microscopy of the oxidized Illinois #6 revealed colonization of the surface by thePenicillium sp., production of conidia, and erosion of the coal surface. The average molecular weight (MW) of Illinois #6 degraded by the fungus and base-solubilized was approximately 1000 Da. The average MW for base-solubilized Illinois #6 that was not exposed to the fungus was 6000 Da, suggesting solubilizing mechanisms other than base catalysis. A spectrophotometric assay to quantify the microbial conversion of biosolubilized coal was developed. Standard curves were constructed based on the absorbance at 450 nm of different quantities of microbe-solubilized coal. An acid precipitation step was necessary to remove medium and/or microbial metabolites from solubilized coal to prevent overestimation of the extent of coal biosolubilization. Furthermore, the absorption spectra for different coal products varied, necessitating construction of standard curves for individual coals.     

Survival of Rhizobium phaseoli in Coal-Based Legume Inoculants

The long-term survival of Rhizobium phaseoli strains 127K17, 127K26, and 127K35 in legume inoculants prepared with eight different coals (one strain and one coal per inoculant) was studied. The coals used were Pennsylvania anthracite, bituminous coals from Illinois, Pennsylvania, and Utah, lignite from North Dakota and Texas, and subbituminous coals from New Mexico and Wyoming; they ranged in pH from 4.7 to 7.5 All coals, with the exceptions of Illinois bituminous coal and Texas lignite (pH's of 5.0 and 4.7, respectively), supported the growth and survival of all R. phaseoli strains. All coal-based inoculants in which rhizobial viability was maintained had more than 10⁶ rhizobia per g for at least 7 months, and most contained more than 10⁷ rhizobia per g after 12 months. It appears that most coals, regardless of grade or source, may be acceptable carriers for R. phaseoli inoculants.     

露天煤矿煤粉沉降对矿区周边主要植物的生理影响

随着新疆准东煤田开采的日益加剧,露天开采及运输造成的煤粉尘不仅影响区域大气环境,同时在一定程度上影响周边植物的生理生长。为探讨煤粉尘对周边植物的生理影响,以准东露天煤矿周边人工绿化树种白榆、乡土植物梭梭和柽柳为研究对象,通过野外控制实验,探究覆尘、无尘条件下3种植物的光合生理特性的变化。结果表明:煤粉沉降对3种植物的光合生理均产生了明显影响,覆尘叶片在不同光照强度下净光合速率、气孔导度、蒸腾速率均降低,净光合速率柽柳(18.6%)梭梭(28.3%)白榆(30.7%),气孔导度梭梭(24.1%)柽柳(30.9%)白榆(33.2%),蒸腾速率梭梭(16.7%)白榆(18%)柽柳(39.1%);煤尘对白榆、梭梭的影响主要为非气孔因素,对柽柳的影响有待继续研究。因此露天煤矿开采造成的粉尘会在一定程度上影响周边植物的光合生理过程,抑制其生长,长期排放可能会导致植被覆盖度降低。     

>Decolourisation and depolymerisation of solubilised low-rank coal by the white-rot basidiomycete Phanerochaete chrysosporium

Peroxidases secreted by the white-rot basidiomycete Phanerochaete chrysosporium can oxidise a wide range of recalcitrant compounds including lignin and aromatic xenobiotics. Since low-rank coals such as brown coal and lignite retain structural features of the parent lignin, we investigated the possibility that P. chrysosporium is capable of acting on a brown coal, with the production of useful low-molecular-mass compounds. In nitrogen-limiting liquid medium containing 0.03% solubilised Morwell brown coal, P. chrysosporium was found to convert about 85% of the coal after 16 days incubation to a form not recoverable by alkali-washing and acid-precipitation. The modal molecular mass of the residual coal macromolecules was reduced from the initial 65kDa to 32 kDa. Extensive bleaching of the coal coincided with the presence of extracellular lignin peroxidase (LiP) and manganese-dependent peroxidase (MnP), although both LiP and MnP activity were lower in cultures containing coal. These reductions are accounted for by interference with the enzyme assays by solubilised coal and by binding of MnP to precipitated coal. LiP was about eight times more sensitive than MnP to inhibition by solubilised coal. In nitrogen-sufficient medium containing solubilised coal, neither coal modification nor LiP activity were observed, suggesting that LiP is an essential component of the bleaching process.     

Solubilization of coal by an extracellular product fromStreptomyces setonii 75Vi2

Summary Several low-ranked coals were solubilized when placed on the surface of agar cultures ofStreptomyces viridosporous T7A andS. setonii 75Vi2. When grown in submerged cultureS. setonii 75Vi2 produced an extracellular component that was capable of solubilizing coals. The extracellular coal solubilizing component had a molecular weight of <10000 and was heat stable since, after 1h at 121°C, only 30–40% of the activity was lost. Treatment with any of three proteases also appeared to be ineffective in decreasing activity. These results suggest that coal solubilization byS. setonii 75Vi2 is nonenzymatic.Research supported by the Fossil Energy Advances Research and Technology Program, managed by the Pittsburg Energy Technology Center, U.S. Department of Energy, under Contract No. DE-AC05-840R21400 with Martin Marietta Energy Systems, Inc.     

SOURCE PLANTS FOR CARBONIFEROUS MICROSPORES: LYCOSPORA FROM PERMINERALIZED LEPIDOSTROBUS

Permineralized Lepidostrobus specimens were examined from the Westphalian A of England, Westphalian B (Breathitt) of the Appalachian Basin, and the Westphalian D (Desmoinesian) of the Illinois Basin and Western Interior Coal Region to correlate species of the dispersed spore genus Lycospora with source cones and cone types (“species”) with parent plants. Based on 51 specimens of Lepidostrobus oldhamius sensu Balbach, three cone types, producing three species of Lycospora, are recognized. Lycospora pellucida occurs in smaller diameter cones with a prominently coronate stele; cones are associated with Lepidophloios harcourtii from the Westphalian A-B. Lycospora pusilla occurs in larger diameter cones with a small “pith” containing elongate, sclerified, tanniferous cells from Westphalian D coal balls. Secretory cells also are present in sporophyll trace sheaths and in subarchesporial parenchyma pads in sporangia. The suggested plant source is Lepidodendron hickii. Lycospora granulata occurs in large cones with a large “pith” composed of thin-walled parenchyma; no secretory cells were observed in the “pith,” sporophyll traces, or subarchesporial pads. These cones are associated with Lepidophloios hallii from the Westphalian D. Association of Lycospora species with source lycopods increases the possibilities of paleoecological interpretation using microspores from Carboniferous coals.     

Nitrogen conversion under rapid pyrolysis of two types of aquatic biomass and corresponding blends with coal

Rapid pyrolysis of two types of aquatic biomass (blue-green algae and water hyacinth), and their blends with two coals (bituminous and anthracite) was carried out in a high-frequency furnace. Nitrogen conversions during rapid pyrolysis of the two biomass and the interactions between the biomass and coals on nitrogen conversions were investigated. Results show that little nitrogen retained in char after the biomass pyrolysis, and NH₃ yields were higher than HCN. During co-pyrolysis of biomass and coal, interactions between biomass and coal decreased char-N yields and increased volatile-N yields, but the total yields of NH₃ + HCN in volatile-N were decreased in which HCN formations were decreased consistently, while NH₃ formations were only decreased in the high-temperature range but promoted in the low-temperature range. Interactions between blue-green algae and coals are stronger than those between water hyacinth and coal, and interactions between biomass and bituminous are stronger than those between biomass and anthracite.     

Does the sequence of plant dominants affect mycorrhiza development in simulated succession on spoil banks?

The aim of this field study was to examine how the development of arbuscular mycorrhizal fungi (AMF) on coal mine spoil banks is affected by the presence of plants with different mycorrhizal status. A 3-year trial was conducted on the freshly created spoil bank Vršany, North-Bohemian coal basin, the Czech Republic. Three plant species – non-mycotrophic annual Atriplex sagittata, highly mycotrophic annual Tripleurospermum inodorum (both dominants of early stages of succession) and facultatively mycotrophic Arrhenatherum elatius (a perennial grass species of the later stage of succession) – were planted on 1 m² plots over 3 years in different sequences that simulated the progress of succession on spoil banks. The development of AMF populations was monitored by evaluation of mycorrhizal colonization of plant roots and by measurement of the mycorrhizal inoculation potential (MIP) of soil. These two parameters were compared between plots inoculated with the mixture of three AMF isolates – Glomus mosseae BEG95, G. claroideum BEG96 and G. intraradices BEG140 – (“inoculated plots”) and plots exposed only to natural dispersal of AMF propagules (“uninoculated plots”). Highly colonized roots of plants together with a high MIP of soil in uninoculated plots were already found at the end of the first season, indicating rapid natural dispersal of AMF propagules. Root colonization of facultatively mycotrophic and non-mycotrophic plants in later years was affected by the mycorrhizal status of the previous plant species. The MIP of soil continuously increased throughout the experiment; in uninoculated plots, the MIP was temporarily decreased if plant species of higher mycotrophy were replaced by species of lower mycotrophy. The results lead to the conclusion that AMF colonize freshly formed sites very quickly and reproduce or accumulate in the soil, which leads to increasing MIP values. However, this infective potential can be decreased if non-mycotrophic plants predominate on the site.     

煤炭井工开采对干旱荒漠区植被动态变化的影响

我国西北干旱荒漠区生态环境脆弱,煤炭开采活动严重地破坏植被和影响生态环境,而煤炭井工开采对干旱荒漠区植被动态变化的影响尚不明确。以灵武市为例,采用遥感技术和野外实地调查相结合的方法,分析2000-2019年间煤炭井工开采对植被动态变化的影响。结果表明：灵武市的植被主要以沙蒿（Artemisia salsoloides）、柠条（Caragana korshinskii）和芨芨草（Achnatherum splendens）等荒漠植物为主;2000-2019年间,植被覆盖度（FVC）和绿度变化率（GRC）表明灵武市植被整体呈现改善趋势;归一化植被指数（NDVI）与年降水量（P）和年平均风速（S）等气象因子显著相关,表明气候因子对区域植被动态变化起主要作用;煤炭开采区侵占草地和灌丛面积,使得土地利用类型发生变化,生态环保政策的实施对于区域土地利用类型的变化和植被改善具有重要作用。实地调查分析表明煤炭开采改变了矿区植物群落结构,植被盖度和物种多样性指数均在煤炭开采后1-4a呈下降趋势,5-9a为上升趋势,10a自然恢复后与对照区的变化趋势一致,说明在自然条件下煤炭开采区植被恢复经历了退化期、改善期和初步恢复期等过程。这些研究结果为西北干旱荒漠区煤炭井工开采矿区植被恢复和生态环境建设提供了理论基础。     

Coal methanogenesis: a review of the need of complex microbial consortia and culture conditions for the effective bioconversion of coal into methane

Microbial biodegradation of coal into low-molecular-weight compounds such as methane has been extensively researched in the last two decades because of the underlying environmental and industrial applications of this technique as compared to the chemical and physical methods of coal conversions. However, the irregular structure of coal and the need for complex microbial consortia under specific culture conditions do not make this biotransformation an ideal process for the development of anaerobic bioreactors. The most abundant species in a methanogenic culture are acetoclastic and hydrogenotrophic methanogens which utilize acetate and H₂+CO₂, respectively. Medium- to low-rank coals such as high-volatile bituminous, sub-bituminous and lignite are more promising in this bioconversion as compared to semi- and meta-anthracite coals. While covering the details of the ideal culture conditions, this review enlightens the need of research setups to explore the complex microbial consortia and culture conditions for maximum methane production through coal methanogenesis.     

Influence of coal source and treatment upon indigenous microbial communities

Summary The leaching of six Eastern coals was investigated using experimental coal columns subjected to simulated leaching events. Measurements of CO₂ assimilation and specific enrichment cultures indicated that the microbial communities of all leachates were dominated by iron- and sulfur-oxidizing chemoautotrophic bacteria. Comparison of CO₂ assimilation rates in leachates and core samples of leached coal indicated that most chemoautotrophs remained within coal columns during leaching. Mean numbers of chemoautotrophic bacteria in leachate samples were correlated with concentrations of dissolved iron and sulfate. Leachates from unwashed, run-of-mine coals contained more chemoautotrophs and more iron and sulfate than did leachates from washed, final product coals. After several leachings, the ratio of sulfur oxidizers to iron oxidizers tended to increase. These data suggest that the chemoautotrophic community of final product coals may be pyritelimited. Aerobic heterotrophs constituted a minor component of the microbial community in leachates from the six coals and their abundance and metabolic activity were apparently not influenced by the beneficiation history of the coal. Changes in rates of acetate metabolism may have been related to microbial succession within the heterotrophic community of coal columns. In all leachates, rates of tritiated methylthymidine assimilation were correlated with rates of acetate incorporation but not with CO₂ assimilation, even though autotrophs dominated the microflora. Thus, thymidine assimilation rates appear to reflect activities or growth of mainly heterotrophic microorganisms in leachate.     

Paleogene coal-forming plants of the Zeya-Bureya Basin (Amur River Region)

Coal-forming plants from the Early Paleogene coal mines of the Zeya-Bureya Basin (Amur River Region) were studied. Dispersed cuticles of taxodialeans were obtained from coals for the first time in this region. Since the Danian, taxodialeans became dominant elements in peat-forming environments and provided abundant plant material that transformed into coal.     

Phyto-bioconversion of hard coal in the Cynodon dactylon/coal rhizosphere

Fundamental processes involved in the microbial degradation of coal and its derivatives have been well documented. A mutualistic interaction between plant roots and certain microorganisms to aid growth of plants such as Cynodon dactylon (Bermuda grass) on hard coal dumps has recently been suggested. In the present study coal bioconversion activity of nonmycorrhizal fungi was investigated in the C. dactylon/coal rhizosphere. Fungal growth on 2% Duff-agar, gutation formation on nitric acid treated coal and submerged culture activity in nitrogen-rich and -deficient broth formed part of the screening and selection of the fungi. The selected fungal isolates were confirmed to be found in pristine C. dactylon/coal rhizosphere. To simulate bioconversion, a fungal aliquot of this rhizosphere was used as inoculum for a Perfusate fixed bed bioreactor, packed with coal. The results demonstrate an enhanced coal bioconversion facilitated by low molecular weight organics and the bioconversion of coal may be initiated by an introduction of nitrogen moieties to the coal substrate. These findings suggest a phyto-bioconversion of hard coal involving plant and microbes occurring in the rhizosphere to promote the growth of C. dactylon. An understanding of this relationship can serve as a benchmark for coal dumps rehabilitation as well as for the industrial scale bioprocessing of hard coal.     

Lignite Biosolubilization by Bacillus sp. RKB 2 and Characterization of its Products

Abstract

Nowadays, the advancements of coal microbiology and biotechnology have been highly emphasized, providing leading-edge approaches in sustainable development of agriculture and the protection of the environment. The biosolubilization of low-rank coals, such as lignite and leonardite is a promising technology for converting these sedimentary rocks into valuable products. In this study, the process involved in lignite biosolubilization by Bacillus sp. RKB 2 was investigated. The biotransformed lignite and the produced humic substances were determined in vitro in a liquid medium and on a solid matrix. The bacterial strain was isolated from untreated Kazakhstani lignite and was shown to be capable of effectively solubilizing and transforming lignite (5% w/v). Fourier Transform Infrared (FTIR) and UHPLC-QqQ-MS/MS analyses were performed to examine the solubilization products and lignite humic substances processed by bacteria. The absorption peaks of FTIR showed the diverse nature of the bacteria-induced humic substances, and the vast majority of intense peaks detected are mainly below an m/z of 1000?Da (liquid chromatography-mass spectrometry [LC-MS] (QqQ)). Data analysis concluded that our isolate could depolymerize lignite and form bio-humic substances. Due to its ability to solubilize lignite Bacillus sp. RKB 2 may be useful in the coal-bed for in situ bioutilization of low-rank coal.     

Bioconversion of coal,lignin, and dimethoxybenzyl alcohol byPenicillium citrinum

Summary Bioconversion of alkali-soluble coal, sulfonated lignin, and dimethoxybenzyl alcohol (DMBA) byPenicillium citrinum was investigated with respect to the effects of (1) these compounds on growth and metabolism, and (2) the organism on the chemical nature of coal and DMBA. Alkali-soluble coal caused a slight enhancement of grwoth and metabolism; DMBA and lignin partially inhibited growth and metabolism. Both whole cells and cell-free extracts were capable of oxidation of DMBA to dimethoxybenzaldehyde. Whole cells demonstrated the capability of modifying alkali-soluble Beulah Zap and Ugljevik lignite coals by producing compounds that were of lower and higher molecular weight than the original coal. In vivo conversion of alkali-soluble Ugljevik coal resulted in a substantial decrease in the sulfur content of the coal (52% decrease). Cell-free extracts were able to degrade alkali-soluble Ugljevik lignite coal. The results suggest a potential usefulness of this microorganism for coal bioprocessing.Nomenclature A light absorption - DMBA 3,4-dimethoxybenzyl alcohol - HPSEC high performance size exclusion chromatography - MW molecular weight     

大柳塔长焰煤中灰分和无机矿物对生物产气的影响

【目的】以不同密度等级大柳塔长焰煤作为产气底物,前期驯化培养厌氧菌群进行生物模拟产气实验,研究不同密度等级煤中的灰分和无机矿物对生物产气的影响。【方法】利用小浮沉将大柳塔长焰煤分成不同密度等级的煤样,采用工业分析、XRD、XRF分析小浮沉处理得到煤样的理化性质,利用这些煤样进行生物产气模拟实验,以甲烷产量作为评价指标,分析不同密度等级煤样中灰分对产气的影响。最后,通过添加几种标准矿物方式比较了煤中无机矿物对生物产气的可能影响。【结果】不同密度等级煤样中灰分对产气量存在一般显著影响(P=0.035),且灰分与甲烷含量呈负相关关系,其灰分中的无机矿物如高岭土、菱铁矿、氧化亚铁镁等的积累对产气有抑制作用。不同矿物配比产气实验证实低含量的粘土矿物促进甲烷的生成,高含量的粘土矿物抑制产气。【结论】不同密度等级煤中的灰分对生物产气存在一般显著的影响,高灰分煤的产气量低,而低灰分煤的产气量高。