基于GET技术的混合厌氧发酵产甲烷研究

GET是一种国外新研发的在田间利用秸秆进行甲烷生产的技术。为了推动该技术在我国的实际应用,本研究以稻草与牛粪为发酵原料,水稻田土壤作为基底,采用正交试验法,测试了温度、发酵基质配比（稻草∶牛粪）和秸秆腐熟剂浓度对混合厌氧发酵产气量的影响。结果表明：各处理因子的产气动态呈“S”型增长模式,但在快速增长时间和增长幅度上有一定区别。30 ℃和40 ℃处理的累积产气量显著高于10 ℃和20 ℃处理的累积产气量;发酵基质配比以1∶1处理的累积产气量最大,分别比1∶0（对照）、1∶2和1∶3配比高34.12%、20.39%和21.78%;秸秆腐熟剂浓度的产气效果在实验范围内随着浓度的增加而增加。厌氧发酵过程中各实验因子下挥发性脂肪酸（VFAs）含量和pH在20 d左右分别有一个高峰和低谷值且都有较大差异,各因子主要是通过在这个时期对厌氧发酵进程的影响而对整个产气量产生影响。综合分析显示稻草厌氧发酵产生物甲烷的最优组合为：温度30 ℃、发酵基质配比1∶1、秸秆腐熟剂浓度8%。     

碳氮比对模拟水稻秸秆田间厌氧发酵系统甲烷生产和秸秆降解的影响

利用自制的厌氧发酵装置模拟田间厌氧发酵产生物甲烷的条件,通过添加不同量的尿素,研究C/N对水稻秸秆厌氧发酵产生物甲烷和秸秆降解的影响。设置了C/N为15∶1、20∶1、25∶1、30∶1和对照(53∶1)等五个处理,测定各处理的产气量、甲烷含量和秸秆木质纤维素含量。结果表明C/N对厌氧发酵产气进程有一定影响,峰值出现的时间、峰值的大小在不同处理之间均有差异:以C/N为25∶1的处理峰值出现的最早,较对照早5 d出现;以C/N为20∶1的峰值最高,较对照提高了27. 55%。C/N对厌氧发酵累积产气量和累积甲烷产量有显著影响,以C/N为20∶1的处理累积产气量(4 235. 00 mL)和累积甲烷产量(1526. 34mL)最高,相比对照组的累积产气量和累积甲烷产量分别提高了30. 78%和51. 31%。C/N对水稻秸秆降解也有显著影响。以20∶1和25∶1处理对秸秆降解的效果最好,其中20∶1处理的总降解率、纤维素降解率和半纤维素降解率分别达到了51. 33%、55. 31%和53. 05%,25∶1处理分别为50. 00%、53. 08%和49. 42%。上述结果表明通过添加尿素调节C/N能够提高水稻秸秆厌氧发酵的产生物甲烷效率和促进秸秆降解,以C/N为(20～25)∶1处理的促进效果最好。     

添加剂对水稻秸秆实地混合厌氧发酵产甲烷系统的作用

为提高水稻秸秆利用效率,改进水稻秸秆实地混合厌氧发酵产甲烷技术,本研究开展了添加剂在混合厌氧发酵系统的应用研究。试验选取水稻秸秆和猪粪作为发酵原料,通过分别添加不同的浓度的吐温20和腐植酸,测定甲烷的产气量和浓度、秸秆的降解和土壤肥力的变化,以揭示添加剂类型及其浓度对水稻田实地甲烷生产系统的影响。结果表明:添加剂的掺入并未影响产气的动态变化趋势,但显著地促进产气和提高产气浓度,整体效果由高到低为腐植酸吐温20对照,其中经腐植酸处理的产气量和产气浓度对照相比分别提高了50. 73%和24. 55%。添加剂的掺入有利于水稻秸秆纤维素和半纤维素的降解,但对木质素没有显著影响;其中以0. 15 g/L腐植酸和0. 30 g/L吐温20的降解率最高,相较于对照其纤维素降解率均提高了22. 11%,半纤维素降解率分别提高了107. 13%和98. 39%。添加剂的掺入能显著增加土壤肥力,以0. 15 g/L腐植酸和0. 30 g/L吐温20处理水平的效果最优,相较于对照,其土壤有机质分别增加了29. 63%和23. 72%,全氮分别增加了52. 32%和42. 38%,全磷分别增加了83. 33%和57. 14%。     

降解菌系和助腐剂对不同还田方式下水稻秸秆降解特征的影响

基于田间小区试验,设置两种秸秆还田处理(以秸秆混土处理模拟旋耕还田,以秸秆不混土处理模拟秸秆沟埋还田),采用尼龙网袋法,通过测定秸秆降解率及秸秆半纤维素、纤维素和木质素的含量,研究两种还田方式下施用降解菌系和助腐剂后水稻秸秆的降解规律。结果表明:在试验前5个月,不混土处理的秸秆降解率始终高于混土处理4.5%～11.6%;至第12个月,混土处理秸秆降解率显著高于不混土处理4.9%～13.0%。在前2个月,各处理水稻秸秆降解较快,秸秆降解率均超过50%,此后秸秆降解进入缓慢期。混土处理中,降解菌系和助腐剂仅能在第1个月提高秸秆的降解效率,此时秸秆降解率分别比对照组高11.0%和10.2%(P0.01);不混土处理中,降解菌系在1～5个月内显著提高秸秆降解效率4.3%～9.7%,助腐剂在1～12个月显著提高秸秆降解效率7.3%～14.4%。试验至第5个月时,喷施助腐剂的秸秆半纤维素、纤维素和木质素的降解率分别达79.1%、80.2%和34.7%,且与喷施菌系处理差异不显著。综上所述,秸秆降解菌系和秸秆助腐剂均能提高水稻秸秆的降解效率,且不混土处理的施用效果好于混土处理。     

应用混料实验设计制备秸秆复合降解菌剂

农业秸秆类废弃物含有大量木质纤维素,该类物质结构稳定,不易降解,为秸秆的合理利用带来诸多困难。本实验尝试利用混料实验设计对筛选出可以共同培养的五种木质纤维素降解菌的配比进行研究,寻求复合发酵降解剂各组分的最佳配比,并分析发酵产品得到适用于不同发酵目的的菌剂。通过对发酵产品中木质素和纤维素降解率及还原糖的含量的分析建立模型,分析预测纤维素降解率最高为35.75%,木质素降解率最高为27%,还原糖含量最高为3.39mg/g。通过优化得出发酵菌剂最优配比为枯草芽胞杆菌12.1%,克鲁斯假丝酵母10%,地衣芽胞杆菌27.2%,变色栓菌10.6%,黄孢原毛平革菌40%。对应三指标的实测值为：35.47%,26.41%和2.37mg/g。     

1株来自瘤胃液菌株RB1在降解秸秆中的作用

木质纤维素中木糖残基大量乙酰化,导致半纤维素的降解受阻。半纤维素支链水解断裂,可以解除其空间位阻效应,利于木质纤维素彻底降解。采用产乙酰酯酶活力较高的菌株RB1降解玉米秸秆和水稻秸秆。研究结果表明,菌株RB1对玉米秸秆和水稻秸秆中半纤维素降解率分别高达53.87%和51.67%。同时该菌株对秸秆中木质素降解率分别达到35.50%和35.01%。该菌株与其他降解纤维素能力较强的菌株共同发酵,会对木质纤维素类物质有更高降解率。该菌株在生物转化木质纤维素类物质方面,具有一定的潜在应用价值。     

大庆地区玉米秸秆分解微生物的筛选及初步研究

目的从大庆地区土壤样品中分离能够降解玉米秸秆的菌株。方法采用刚果红染色法和滤纸片失重法筛选得到具有较高降解纤维素能力的菌株,进一步测定菌株对玉米秸秆中纤维素的降解能力,最后测定温度和p H对菌株生长的影响。结果得到能够降解玉米秸秆的菌株4株,其中4#菌株的纤维素降解能力为23.32%,秸秆失重率为28.86%,相对适宜的培养温度为37℃,p H为6.0-7.0。结论得到一株具有较高秸秆分解能力的大庆地区土壤本源菌。     

秸秆降解菌的筛选及对秸秆的降解效果

作物秸秆作处置不当可能严重影响农村生态环境。目前东北地区的秸秆处置方式主要为直接打碎还田,但秸秆在自然环境中不易腐化,影响春耕。从添加外源微生物促进秸秆原位腐化角度开发新型可培养秸秆降解菌,具有重要意义。本实验通过菌种富集培养、刚果红培养基初筛和滤纸条崩解试验复筛的方法,从腐烂的秸秆和牛肠道中分离筛选潜在的高效纤维素降解菌,测定其最适生长温度和pH,在液态发酵培养条件下考察菌株实际降解能力,共获得具有较高玉米秸秆降解能力的降解菌5株。5种菌株的生长峰值均出现在温度20～30℃,pH值7.5～8.5范围内。液态发酵培养15天后,秸秆失重率为菌株NX9(53.88%)NF6(51.36%)JF3(46.97%)JZ8(45.2%)JX4(35.79%)CK(23.88%)。其中,菌株NX9(温度30℃、pH 7.5)对秸秆半纤维素和木质素的降解能力最强,15天降解率分别为48%和37.7%;筛选出的NF6和JF3属于耐冷微生物,特别是菌株NF6在4℃条件下也能生长繁殖,为北方开展"外源微生物促进秸秆原位腐化"技术提供了基础。     

秸秆还田对稻田土壤水稳性团聚体养分及其生态化学计量比的影响

以福州平原早、晚稻田为研究对象,分析了秸秆还田对早、晚稻田土壤不同粒级水稳性团聚中TC、TN、TP含量及其计量比的影响。结果表明:秸秆还田之后,早稻土壤各粒级水稳性团聚体中,TC、TN、TP含量以及TC∶TN均不同程度增加,且在1~2 mm粒级水稳性团聚体中,TC、TP含量及TC∶TN分别增加42.3%、22.2%和29.1%(P0.05);各粒级水稳性团聚体中,TN∶TP均不同程度减小,TC∶TP则无明显变化规律;秸秆还田后,晚稻土壤1~2 mm粒级水稳性团聚体中TC、TP显著降低(P0.05),而各粒级水稳性团聚体中TC∶TN基本无变化,TC∶TP和TN∶TP则在1~2 mm粒级水稳性团聚体中增大,其余粒级团聚体中减小;从团聚体粒级角度看,早、晚稻土壤中TC、TN、TP主要分布在水稳性大团聚体中(0.25~2 mm粒级),团聚体粒级越小,养分含量越少;秸秆还田后,晚稻土壤水稳性大团聚体对土壤养分贡献率增大,水稳性微团聚体(0.25 mm粒级)养分贡献率则降低;总体上,早、晚稻田中,1~2 mm粒级水稳性团聚体对秸秆还田响应显著;秸秆还田对福州稻田水稳性团聚体中养分含量有影响,对其化学计量比的影响则不明显。     

一株玉米秸秆纤维素降解放线菌的筛选、鉴定及酶学特性研究

为解决玉米秸秆固废污染和秸秆资源有效利用问题，采用刚果红染色法（水解圈法）和3，5-二硝基水杨酸（DNS）法从玉米秸秆还田土壤中筛选到一株纤维素降解菌，并对该微生物进行生理生化和分子生物学鉴定，发现该菌株降解纤维素效果较好，经鉴定该菌株为纤维素链霉菌（Streptomyces cellulosae），命名为SJS-15，并对该菌株的酶学特性及纤维素降解能力进行了初步研究。结果表明，菌株SJS-15在发酵培养基中的纤维素酶活（CMC）峰值为30.5 U/mL，最适反应pH为6.0，滤纸酶活（FPA）峰值为25 U/mL，最适反应pH为8.0，两种酶均能在温度20~60 ℃，pH 4.0~10.0范围内保持较高酶活性。纤维素分解实验表明菌株SJS-15对玉米秸秆和滤纸有分解能力，40 d时对玉米秸秆降解率为35.6%（质量分数，下同），对滤纸降解率为18.6%。扫描电镜结果显示经菌株处理的玉米秸秆较对照有明显降解痕迹。菌株SJS-15具有良好的抗逆性和玉米秸秆纤维素分解能力，可作为玉米秸秆还田和堆肥发酵的高效菌株进行进一步研究。     

Bioconversion of wheat straw and wheat straw components into single-cell protein.

Several fungi (Aspergillus niger, A. terreus, Cochliobolus specifer, Myrothecium verrucaria, Rhizoctonia solani, Spicaria fusispora, Penicillium sp., and Gliocladium sp.) were isolated from decomposing wheat straw and tested for their ability to utilize whole straw and its components, holocellulose (hemicellulose and cellulose) and cellulose, for the production of single-cell protein (SCP). It was found that C. specifer was the most efficient fungus for protein synthesis with the three substrates. Using potassium nitrate as N source in mixtures of 0.04 g N/g substrate (0.04% wt./vol.) at pH 4.5, it was found that incubation periods of 3, 4, and 5 days were optimal for protein production on cellulose and holocellulose fractions, and whole straw, respectively. Whole native straw was found to be the most recalcitrant to bioconversion into SCP; however, protein production was almost doubled when the lignin component was removed using a mixture of sodium chlorite and acetic acid.     

土壤呼吸对秸秆与秸秆生物炭还田的响应及其微生物机制

土壤呼吸释放CO_2是温室气体排放的重要途径之一,减少土地利用中温室气体排放、增强土壤碳汇聚能力对于减缓全球温室效应具有重要意义。生物炭具有改善土壤理化性质、增加作物产量、调节土壤微生物性质等特性。本研究采用室外盆栽的方式,以地肤草为目标植物,研究了芦苇、水稻、互花米草三种农林秸秆及其秸秆生物炭还田对土壤的改良效应,以及对土壤呼吸的影响及其微生物机制。结果表明:秸秆及其秸秆生物炭均可改善土壤肥力,促进植物生长,且生物炭改良效果略好于秸秆直接还田。但秸秆生物炭还田的土壤呼吸显著低于秸秆直接还田,其中芦苇生物炭最低。秸秆直接还田可促进土壤β-糖苷酶、脱氢酶和活性微生物量等微生物活性指标,从而促进土壤呼吸,增加土壤CO_2的释放,而生物炭还田对土壤微生物活性无显著的促进作用,反而有一定的抑制作用,这可能是由于生物炭中易降解有机物含量很低,可降解性较低的缘故。     

Biomethanation of rice and wheat straw

When rice or wheat straw was added to cattle dung slurry and digested anaerobically, daily gas production increased from 176 to 331 l/kg total solids with 100% rice straw and to 194 l/kg total solids with 40% wheat straw. Not only was methane production enhanced by adding chopped crop residues but a greater biodegradability of organic matter in the straws was achieved.The authors are with the Microbiology and Molecular Genetics Unit, Tata Energy Research Institute, 158 Jor Bagh, New Delhi 110 003, India     

秸秆及秸秆黑炭对小麦养分吸收及棕壤酶活性的影响

通过小麦盆栽试验,研究了玉米秸秆及其秸秆黑炭施加对小麦养分吸收利用和棕壤酶活性的影响。试验设对照(CK),黑炭(B),秸秆还田(S),尿素(U),尿素+黑炭(UB)及尿素+秸秆还田(US)6个处理,各处理3次重复。结果表明:无氮肥施入下,B处理较CK和S处理籽粒产量显著提高99.4%和77.7%,小麦地上部氮、磷、钾吸收累积量分别显著提高94.1%—140.9%,55.4%—66.3%和53.1%—72.6%;有氮肥施入下,UB和US处理较U处理提高籽粒产量8.2%—8.8%,小麦地上部氮、磷、钾吸收累积量分别显著提高14.3%—27.8%,19.6%—30.9%和24.4%—40.9%。秸秆及秸秆黑炭施加处理的氮素利用率显著提高21.4%—41.7%。黑炭施加显著提高土壤中有机碳、NH+4-N、NO-3-N和速效钾含量;在施氮条件下,秸秆还田显著提高土壤中NO-3-N含量;秸秆及黑炭施加对有效磷含量无显著影响。秸秆还田显著提高了土壤脱氢酶、过氧化氢酶、脲酶和中性磷酸酶活性;施加黑炭也明显提高了土壤脱氢酶和脲酶活性,但抑制过氧化氢酶和中性磷酸酶活性。土壤脲酶活性与土壤有机碳、无机氮含量呈显著正相关,表明土壤酶可反映土壤肥力水平。     

Cultivation ofPleurotus florida mushroom on rice straw and biogas production from the spent straw

Rice straw, used as a substrate for three successive crops of the fruiting bodies ofPleurotus florida having 22% protein, had less cellulose but more nitrogen and ash than the original straw.In vitro digestibility using bacterial cellulase released 4.3-fold more reducing sugars per g cellulose from spent straw than from plain straw. There was 8-fold increase in biogas production from the spent straw compared with the original when used either in 31 (w/w) or 11 (w/w) combination with cattle dung.

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Résumé La paille de riz, utilisée comme substrat pour trois récoltes successives de corps fruités dePleurotus florida, à 22% de protéines, contenait moins de cellulose mais plus d'azote et de cendres que la paille originelle. La digestionin vitro par une cellulase bactérienne, relarguait 4.3 fois plus de sucres réducteurs par g de cellulose à partir de la paille résiduaire qu'à partir de la paille originelle. On observe un accroissement de 8 fois dans la production de biogas à partir de la paille résidualle par rapport à la paille originelle lorsque cellesci sont utilisées en combinaison avec la bouse de vache dans les proportions soit de 31 soit de 11 (p/p).

     

The feeding value of ammoniated flax straw,wheat straw and wheat chaff for beef cattle

Two experiments were conducted to assess the feeding value of ammoniated and untreated flax straw, wheat straw and wheat chaff in comparison to a mixed bromegrass/alfalfa hay. Anhydrous ammonia was applied to the crop residues at the rate of 35 kg t⁻¹ dry matter. In the first experiment, the effect of ammoniation on crude protein, acid detergent fibre (ADF), neutral detergent fibre (NDF) and acid detergent lignin (ADL), digestible organic matter in vitro and in vivo (DOM%), ADF and NDF digestibility of the crop residues was determined. In the second experiment, ammoniated flax straw, ammoniated wheat straw, ammoniated and untreated wheat chaff, each supplemented with barley, were compared to bromegrass/alfalfa hay as feed sources for wintering beef cows.Ammoniation increased the crude protein content of the crop residues ∼2-fold. Wheat straw DOM in vitro and in vivo was not increased by ammoniation. Ammoniation increased the DOM in vitro of wheat chaff from 36.3 to 46% and flax straw from 35.2 to 46.3%. The DOM in vivo increased from 53.3 to 63.4% (P < 0.05) for wheat chaff and from 33.9 to 58.4% (P < 0.05) for flax straw following ammoniation. Digestibility of ADF increased from 9.9 to 43.9% (P < 0.05) and of NDF from −0.6 to 37.9% (P < 0.05) in flax straw with ammoniation. Non-significant increases in ADF and NDF digestibility were observed for all other crop residues. Lignin content was not changed in the crop residues by ammoniation.In the winter feeding trial, young cows gained more weight than older cows (P < 0.05). Average daily gains of cows were greatest for hay followed by ammoniated flax straw, ammoniated chaff, untreated chaff and ammoniated wheat straw rations (P < 0.05). Increases in backfat in the younger cows was greatest with hay and ammoniated flax straw, followed by ammoniated chaff and ammoniated wheat straw (P < 0.05). Untreated chaff caused no increase in backfat thickness.Ammoniated flax straw (3.2 kg day⁻¹) given with barley (5.6 kg day⁻¹), is similar in feeding value to medium quality bromegrass/alfalfa hay. Furthermore, wheat chaff and ammoniated wheat chaff show good potential as alternatives to hay in winter feeding.     

Acidogenic and methanogenic fermentation of causticized straw

The effects of pretreatment process variables [straw concentration between 20 and 90 kg volatile solids (VS)/m(3), temperature between 30 and 85 degrees C, and alkaline dosage between 0 and 80 g NaOH/kg VS] on acidogenesis and methanogenesis were investigated. Rates of acidogenesis and methanogenesis were determined using firstorder kinetics, and ultimate acid and methane yields were measured. The acid yield was not affected by pretreatment concentration or temperature, but increased as alkaline dosage increased. The acidogenesis rate was not affected by pretreatment temperature or alkaline dosage, but decreased as the substrate concentration increased. This decrease in the acidogenesis rate was attributed to a decrease in the inoculum: substrate ratio as the substrate concentration increased. The methane yield and methanogenesis rate were not affected by pretreatment substrate concentration or temperature, and both increased with alkaline dosage up to about 40 g NaOH/kg VS, then remained relatively constant above 40 g NaOH/kg VS.     

Fungi of wheat and broad-bean straw composts

13 thermophilic genera and 19 species in addition to one variety of each of M. pulchella and H. grisea were collected from wheat and broad-bean straw composts at 45 °C. In wheat and broad-bean, all thermophilic fungi were completely checked between 4–9 days, and 1–8 days composting when the temperature ranged between 58° and 67 °C, and 58 ° and 70 °C respectively, and reappeared, represented by P. duponti, M. albomyces, T. lanuginosus and S. thermophile, after 9 or 10 days composting when the temperature decreased to 51.5° –54 ° C. Wheat and broad-bean straw composts were analysed biochemically to follow the changes in ethanol and diastase soluble, hemicellulose, cellulose and lignin fractions during composting.     

Fungal colonization and decomposition of cereal straw

The mycofloras of freshly harvested wheat and barley straw were assessed using direct and dilution plating methods and media of high and low water availability. Straw from the standing crop was predominantly contaminated with field fungi, including yeasts, mycelial yeasts and filamentous fungi such as Alternaria, Cladosporium, Epicoccum, Fusarium and Verticillium spp. However, chopped and baled straw had much greater populations of storage fungi, particularly Aspergillus and Penicillium spp., and Wallemia sebi. The dominant colonizers and rate of deterioration of straw held at different levels of water availability were determined over a period of 12 weeks at 15 and 25°C. At 15°C, and −7·0 MPa water potential (=0·95 water activity) Penicillium spp., were predominantly isolated; at −2·8 MPa (=0·98 water activity) Penicillium, Mucor, Rhizopus, Trichothecium and occasionally Fusarium spp. were present. Saturated straw was colonized rapidly by Mucor, Rhizopus, Penicillium, Stachybotrys, Fusarium and sometimes Trichoderma spp. At 25°C the patterns of colonization were different, particularly at 7·0 MPa water potential where Aspergillus spp. were dominant. The percentage dry matter lost over the 12 weeks varied with both temperature and water availability.     

Estimation of metabolisable energy and net energy of rice straw and wheat straw for beef cattle by indirect calorimetry

Two experiments were conducted to estimate the metabolisable energy (ME) and net energy (NE) of rice straw and wheat straw for beef cattle. In each experiment, 16 Wandong bulls (Chinese indigenous yellow cattle) were assigned to 4 dietary treatments in a completely randomised design. Four dietary treatments included one corn silage-concentrate basal diet and three test diets in which the basal diet was partly substituted by rice straw (Exp. 1) or wheat straw (Exp. 2) at 100, 300 and 600 g/kg. Total collection of faeces and urine was conducted for 5 consecutive days after a 2-week adaption period, followed by a 4-d period where gas exchange measurements were measured by an open-circuit respiratory cage. Linear regression equations of rice straw- or wheat straw-associated ME and NE contribution in test diets against rice straw or wheat straw substitution amount were developed to predict the ME and NE values of rice straw and wheat straw. These regression equations resulted in ME and NE values (dry matter basis) of 6.76 and 3.42 MJ/kg for rice straw and 6.43 and 3.28 MJ/kg for wheat straw, respectively. The NE and ME requirement for maintenance of Wandong cattle fed a straw-based diet were 357 and 562 kJ·kg^?0.75·d^?1, respectively. The regression-derived ME and NE have lower standard errors and coefficients of variation than those estimated by any single substitution ratio. Our study found that the regression method based on multiple point substitution is more reliable than the substitution method for energy evaluation of feedstuffs for beef cattle.