Determination of the archaeal and bacterial communities in two-phase and single-stage anaerobic systems by 454 pyrosequencing

2-Phase anaerobic digestion (AD), where the acidogenic phase was operated at 2 day hydraulic retention time (HRT) and the methanogenic phase at 10 days HRT, had been evaluated to determine if it could provide higher organic reduction and methane production than the conventional single-stage AD (also operated at 12 days HRT). 454 pyrosequencing was performed to determine and compare the microbial communities. The acidogenic reactor of the 2-phase system yielded a unique bacterial community of the lowest richness and diversity, while bacterial profiles of the methanogenic reactor closely followed the single-stage reactor. All reactors were predominated by hydrogenotrophic methanogens, mainly Methanolinea. Unusually, the acidogenic reactor contributed up to 24% of total methane production in the 2-phase system. This could be explained by the presence of Methanosarcina and Methanobrevibacter, and their activities could also help regulate reactor alkalinity during high loading conditions through carbon dioxide production. The enrichment of hydrolytic and acidogenic Porphyromonadaceae, Prevotellaceae, Ruminococcaceae and unclassified Bacteroidetes in the acidogenic reactor would have contributed to the improved sludge volatile solids degradation, and ultimately the overall 2-phase system''s performance. Syntrophic acetogenic microorganisms were absent in the acidogenic reactor but present in the downstream methanogenic reactor, indicating the retention of various metabolic pathways also found in a single-stage system. The determination of key microorganisms further expands our understanding of the complex biological functions in AD process.     

污泥与秸秆共基质中温两相厌氧消化特性

以污泥和秸秆为共基质,以沼气产量、ρ（VFA）（VFA为挥发性脂肪酸）和COD_Cr去除率等为指标,探究污泥与秸秆配比（以COD_Cr计,质量比分别为1:0、1:1、2:1、3:1）对中温两相厌氧消化工艺运行效能的影响,以及最佳配比时SRT（污泥停留时间）对产甲烷相厌氧消化稳态特性的影响.结果表明:与污泥试验组相比,添加秸秆试验组的厌氧消化效能均较好;污泥与秸秆的最佳配比为2:1,该稳定状态下产酸相COD_Cr的去除率最高,为17.5%,ρ（VFA）为752 mg/L;产甲烷相COD_Cr的去除率为33.5%,ρ（VFA）为250 mg/L,产气量为47.7 mL/d,总体运行效能较高.在最佳污泥与秸秆配比（2:1）并设定产甲烷相反应器的SRT为20 d时,稳定状态下产甲烷相各组分的变化情况:COD_Cr去除率为41.20%,ρ（VFA）为238 mg/L,产气量为51.3 mL/d,沼气产率为8.4 mL/（d·g）.研究显示,当控制污泥与秸秆配比为2:1、SRT为20 d时,中温两相厌氧消化工艺运行效果良好.      

Microbial community structures in an integrated two-phase anaerobic bioreactor fed by fruit vegetable wastes and wheat straw

The microbial community structures in an integrated two-phase anaerobic reactor（ITPAR）were investigated by 16 S r DNA clone library technology. The 75 L reactor was designed with a 25 L rotating acidogenic unit at the top and a 50 L conventional upflow methanogenic unit at the bottom, with a recirculation connected to the two units. The reactor had been operated for 21 stages to co-digest fruit/vegetable wastes and wheat straw, which showed a very good biogas production and decomposition of cellulosic materials. The results showed that many kinds of cellulose and glycan decomposition bacteria related with Bacteroidales,Clostridiales and Syntrophobacterales were dominated in the reactor, with more bacteria community diversities in the acidogenic unit. The methanogens were mostly related with Methanosaeta, Methanosarcina, Methanoculleus, Methanospirillum and Methanobacterium; the predominating genus Methanosaeta, accounting for 40.5%, 54.2%, 73.6% and 78.7% in four samples from top to bottom, indicated a major methanogenesis pathway by acetoclastic methanogenesis in the methanogenic unit. The beta diversity indexes illustrated a more similar distribution of bacterial communities than that of methanogens between acidogenic unit and methanogenic unit. The differentiation of methanogenic community composition in two phases, as well as pH values and volatile fatty acid（VFA） concentrations confirmed the phase separation of the ITPAR. Overall, the results of this study demonstrated that the special designing of ITPAR maintained a sufficient number of methanogens, more diverse communities and stronger syntrophic associations among microorganisms, which made two phase anaerobic digestion of cellulosic materials more efficient.     

Response of anaerobes to methyl fluoride, 2-bromoethanesulfonate and hydrogen during acetate degradation

To use the selective inhibition method for quantitative analysis of acetate metabolism in methanogenic systems,the responses of microbial communities and metabolic activities,which were involved in anaerobic degradation of acetate,to the addition of methyl fluoride(CH3F),2-bromoethanesulfonate(BES)and hydrogen were investigated in a thermophilic batch experiment.Both the methanogenic inhibitors,i.e.,CH3F and BES,showed their effectiveness on inhibiting CH4 production,whereas acetate metabolism other than acetoclastic methanogenesis was stimulated by BES,as reflected by the fluctuated acetate concentration.Syntrophic acetate oxidation was thermodynamically blocked by hydrogen(H2),while H2-utilizing reactions as hydrogenotrophic methanogenesis and homoacetogenesis were correspondingly promoted.Results of PCR-DGGE fingerprinting showed that,CH3F did not influence the microbial populations significantly.However,the BES and hydrogen notably altered the bacterial community structures and increased the diversity.BES gradually changed the methanogenic community structure by affecting the existence of different populations to different levels,whilst H2 greatly changed the abundance of different methanogenic populations,and induced growth of new species.     

两相厌氧处理有机废水快速启动试验研究

针对试验设计的两相厌氧反应器(TPAD)进行启动研究,寻求快速启动的方法. 试验采用人工配水,以淀粉、葡萄糖作为主要碳源. 采用低负荷启动方式,快速提高进水ρ(COD_Cr),缩短水力停留时间(HRT),使产酸相尽快维持在酸性最佳条件下. 间歇投加粉末状CaO调节产酸相出水pH在7.0±0.3,从而保证产甲烷相在最佳条件下. 经过36 d的启动过程,在产酸反应器和产甲烷反应器中均出现了性能良好的厌氧颗粒污泥. 启动后第36天,当两相厌氧反应器的水力停留时间(HRT)为17.03 h,COD_Cr负荷(VLR)为8.46 kg/(m3·d),水力负荷为0.059 m3/(m2·h)时,系统整体COD_Cr去除率达到最优,为96.66%.      

氨氮浓度对鸡粪中高温甲烷发酵的影响

为探究氨氮浓度对鸡粪中高温甲烷发酵的影响，采用固定水力停留时间（HRT，20d），提高进料总固体浓度（TS，5%、7.5%和10%）的方式增加氨氮浓度，通过265d的长期甲烷发酵试验，比较了不同氨氮浓度条件下鸡粪中高温甲烷发酵效果和污泥的比产甲烷活性.结果显示，TS由5%增至10%，中高温反应器中氨氮浓度由2.1~2.5g/L增至6.1~6.5g/L，对应的比产甲烷活性分别降低了44%和100%，中温反应器中挥发性脂肪酸由0.4g/L增至7.6g/L，甲烷产率由253mL/gTS降至203mL/gTS；高温反应器中挥发性脂肪酸由0.4g/L增至26.1g/L，甲烷产率由181mL/gTS降至18mL/gTS.氨氮浓度对高温甲烷发酵系统的抑制作用更加明显.     

生产性UASB反应器的中温高温交替启动运行过程

处理柠檬酸废水的UASB反应器因所处地区季节气温相差较大,可在中温和高温条件下交替启动运行.反应器在中温条件下启动,串联使用的两级UASB反应器的COD去除率达到77%～86%;当气温升高反应器内水温达到44℃～45℃时,反应器升温进入高温启动运行阶段,两级UASB反应器的COD去除率达到84%～93%;UASB反应器停产38d后,进行了中温再启动,COD去除率达到82%～96%.本文报道了生产性UASB反应器中温、高温交替启动运行的过程,以及中温和高温厌氧颗粒污泥的特性.     

Effect of nitrite on hydrolysis-acidification, biogas production and microbial community in semi-continuous two-phase anaerobic digestion of sewage sludge

Previous study found that the pre-treatment of sewage sludge with nitrite improves the biogas production during the mono/two-phase anaerobic digestion (AD) using batch biochemical methane potential tests. In this study, the effects of nitrite on hydrolysis-acidification, biogas production, volatile solids destruction and microbial composition in semi-continuous two-phase AD of sewage sludge were investigated. The addition of nitrite promotes sludge organic matter solubilization (+484%) and VFAs production (+98.9%), and causes an increase in the VS degradation rate during the AD process (+8.7%). The comparison of biogas production from the acidogenic and methanogenic reactors with or without the addition of nitrite implies that the nitrite has no significant effect on the overall biogas production of two-phase sludge AD process. High-throughput sequencing analysis shows that the microbial communities of bacteria and archaea in two-phase AD reactors significantly changes after the addition of nitrite. Vulcanibacillus (bacteria) and Candidatus Methanofastidiosum (archaea) become the dominant genera in the acidogenic and methanogenic reactors with the nitrite respectively. These findings provide new insights about using nitrite to promote the organic matter degradation of sewage sludge in a semi-continuous two-phase AD system.     

温度对FBAR反应器的运行特性及古菌微生物群落影响

为探讨固定床厌氧反应器（FBAR）在不同温度下的运行特性及微生物群落变化,比较了高温（50℃）、中温（35℃）、低温（4℃）3个温度阶段反应器产甲烷特性及古菌群落变化.结果表明;绝对产气量由大至小依次为高（50℃）、中（35℃）、低温（4℃）,单位负荷产气量依次为中温（2.84L/OLR）,低温（2.5L/OLR）,高温（1.8L/OLR）;甲烷含量依次为低温（74.5%）、中温（63.5%）,高温（57.3%）,不同温度阶段对挥发性有机酸含量变化有一定的影响.克隆文库分析表明：不同温度条件下固定床厌氧反应器内部微生物群落的丰富性存在很大的差异.定量PCR分析表明：甲烷鬃毛菌是中温和高温反应器内的优势菌,低温4℃炭纤维载体和污泥中的优势菌都是甲烷微菌.从能耗、经济效益角度分析低温条件更适合沼气发酵,而主要是以嗜氢产甲烷菌代谢途径为主.     

鸡粪中高温厌氧甲烷发酵产气潜能与动力学特性

采用富含氮素的鸡粪为原料,包括原料鸡粪、鸡粪固相部分和鸡粪液相部分,选取以鸡粪为原料连续稳定运行超过90d的中高温厌氧反应器新鲜出料为接种污泥,在中温（35℃）和高温（55℃）条件下开展动力学和产甲烷潜能试验.采用Gompertz模型、一级动力学模型和两阶段模型对鸡粪中高温累积产甲烷量进行拟合.结果表明,鸡粪中高温甲烷发酵均呈现明显的快速产气期和慢速产气期两阶段特征,快速产气期的动力学常数K₁分别为0.4174和0.2104d^-1,快速产气分别在4.5和6.5d结束,快速产气量占到总产气量的69%和58%.原料鸡粪和液相部分的中温发酵动力学常数（K₁）分别为0.4177和0.2330d^-1,均高于高温的0.1721,0.2214d^-1,发酵产气速率较快.鸡粪固相部分中温发酵的动力学常数为0.1960d^-1,低于液相中温发酵的0.2330d^-1和固相高温的0.2310d^-1,中温条件下,水解过程是限制鸡粪甲烷发酵速率的主要因素之一.鸡粪固体和鸡粪液体高温发酵的动力学常数K分别为0.2310,0.22214d^-1,鸡粪固体发酵产甲烷的速率快于液相部分,水解过程不是限制鸡粪高温发酵产甲烷速率的最主要因素.产甲烷潜能试验表明鸡粪在中温和高温下产甲烷潜能分别为212,177mL/gTS.因此,仅从发酵效率的角度考虑,鸡粪中温发酵比高温发酵的产甲烷潜能更高,产甲烷速率更快.     

厌氧折流板反应器处理糖蜜酒精废水的研究

为探究厌氧折流板反应器(ABR)对糖蜜酒精实际废水的处理效果,该实验采用已成功处理人工模拟糖蜜酒精废水的ABR,研究了该反应器处理糖蜜酒精工业废水过程中COD和SO42-的去除效果,以及各隔室VFA、pH和S2-的分布规律。实验结果表明,反应器处理糖蜜酒精工业废水,在30 d内达到稳定,COD和SO42-负荷分别为4.8 kg/(m3.d)和0.32 kg/(m3.d),COD和SO42-的去除率分别为83%和98%。反应器内各隔室挥发性脂肪酸(VFA)浓度变化规律与pH值变化规律一致,各隔室硫化物(S2-)浓度较低,其中第4、5隔室的S2-浓度低于40 mg/L。反应器内微生物菌群仍能保持处理模拟废水时形成的多相(产酸硫酸盐还原相和生成硫单质产甲烷相)分离特征,保证了ABR对实际工业废水的处理效率。     

碳源对反硝化细菌的反硝化速率和群落结构的影响

为探究碳源类型在反硝化过程中对氮素转化和微生物群落组成的影响,分别建立R1（以C₆H₁₂O₆为碳源）和R2（以CH₃COONa为碳源）反应器,通过分析R1和R2反应器中反硝化过程的氮素转化情况,评价C₆H₁₂O₆和CH₃COONa对脱氮效果的影响,并运用动力学模型对R1和R2反应器中反硝化能力进行评价;同时,采用高通量测序技术表征2种碳源对反应器中微生物群落结构和多样性的影响.结果表明:①运行后期的R1、R2反应器中单位生物量的反硝化速率（以NO₃--N计,下同）分别为8.56、11.26 mg/（g·h）,R1反应器中NO₂--N累积平均值为11.34 mg/L,显著高于R2反应器（0.20 mg/L）,且R1反应器中NH₄+-N累积平均值为6.58 mg/L,是R2反应器（0.65 mg/L）的10.11倍.②反应器中NO₃--N还原过程均符合Haldane模型,其中R1、R2反应器中单位生物量的r_max（最大降解速率）分别为35.61、47.79 mg/（g·h）,表明R2反应器中的反硝化能力强于R1反应器.③微生物经过富集后,其细菌多样性和物种丰度下降,但发挥反硝化作用的微生物相对丰度逐渐增加.R1和R2反应器中共同的优势菌门有Proteobacterias、Bacteroidetes、Firmicutes和Gracilibacters,其在R1反应器中的相对丰度依次为96.14%、2.06%、0.66%和0.47%,在R2反应器中依次为79.75%、6.88%、9.47%和2.13%,优势菌门在不同运行时间的丰度表达上存在消长变化状态.研究显示,C₆H₁₂O₆和CH₃COONa在反硝化过程的氮素转化上存在明显差异,对各类优势菌群的相对丰度有明显影响.      

两级完全混合发酵工艺开发碳源试验研究

采用两级完全混合发酵工艺进行碳源开发试验,研究了水力停留时间(HRT)和污泥停留时间(SRT)对工艺开发碳源效果的影响.研究表明,固定SRT为4d,污泥回流比为1,HRT在32~36h,系统出水的溶解性COD(SCOD)可维持在1090~1180mg/L,且浓缩池泥水分离效果较好,出水的SS保持在50~80mg/L.保持系统的HRT为32h,污泥回流比为1,SRT在4~7d时,可实现厌氧系统中产酸菌与产甲烷菌的分离,系统能够保持良好的产酸效果,此时系统出水的SCOD值基本稳定于980~1179mg/L.碱度可以作为衡量水解酸化系统是否有效运行的重要参数.HRT为32h,SRT为4d的工况下,初沉污泥酸化发酵系统对氨氮和磷的释放率分别为3.88 mgN/gVSS和0.27mgP/gVSS.     

利用硅灰石原位改善厌氧消化微生物电解池的产气特性

厌氧消化-微生物电解池(Anaerobic digestion-Microbial electrolysis cell,AD-MEC)具有有机物降解速度快、降解率高的优点,但其所产沼气中依然存在CO_2含量较高的问题.为降低AD-MEC所产沼气中CO_2的含量,本研究将矿物碳酸化耦合入AD-MEC中,研究添加硅灰石对AD-MEC中CO_2的固定效果.实验结果表明,添加硅灰石可使AD-MEC中CO_2产生量减少40.0%,沼气中CO_2含量从10.0%±1.3%减少到4.5%±1.1%;X射线衍射(XRD)及扫描电镜-能谱(SEM-EDS)分析表明了CaCO_3沉淀的生成,证明硅灰石介导矿物碳酸化固定了AD-MEC中的CO_2.此外,添加硅灰石使Ca~(2+)溶出,缓冲了pH,减轻了厌氧消化产酸阶段对产甲烷菌的抑制,促进了有机物的降解,可溶性化学需氧量(SCOD)去除率提高了11.2%,并使CH_4产量提高18.0%,CH_4产率达到305 mL·g~(-1),沼气中CH_4含量达到95.5%±1.2%.硅灰石的添加实现了AD-MEC中CO_2的原位捕获,同时增益了厌氧消化效果,提高了甲烷产量.     

高温和中温ASBR处理热水解污泥的对比

进行了高温、中温厌氧序批式反应器(ASBR)处理热水解污泥的对比试验研究.在HRT＝10d,总COD(TCOD)容积负荷为5.42 kg/(m³·d) 的条件下,高温、中温ASBR的TCOD去除率分别为56.20%、61.66%,污泥COD的产气率(CH₄)分别为199、219 mL/g.ASBR能有效积累污泥悬浮固体从而保持较高的固体停留时间(SRT),高温、中温ASBR的平均SRT分别为30、37d.同中温ASBR比较,高温ASBR的微生物形态单一、种类少和产甲烷活性较低,因此高温ASBR的处理效率和产气率较中温低.     

热碱解-中温两相厌氧消化工艺运行效能及抗生素抗性基因(ARGs)变化研究

为实现剩余污泥稳定化,并评估剩余污泥的生态环境风险,本研究采用热碱解-中温两相厌氧消化(Thermal-Alkaline Pretreatment-Mesophilic Two Phase Anaerobic Digestion,taMTPAD)工艺处理剩余污泥.考察了热碱解条件及在20、16和10 d 3个不同的总水力停留时间(Total Hydraulic Retention Time,tHRT)下,taMTPAD工艺的运行效果及消化前后抗生素抗性基因(ARGs)的变化.结果表明,对于taMTPAD工艺,当tHRT=10 d时,产酸相的挥发性脂肪酸(VFA)积累量和产甲烷相的日产气量达到最大值,但对ARGs的控制效果最差;tHRT=16 d时反应器的运行效果与tHRT=20 d时接近,但对磺胺类ARGs的控制效果不如tHRT=20 d时;当tHRT=20 d时,挥发性固体(VS)去除率和ARGs的削减效果最好.对比tHRT=10 d时的taMTPAD工艺和MTPAD(两相厌氧消化)工艺,发现对剩余污泥进行热碱预处理能提升产酸相150.32%的VFA产量和产甲烷相89.3%的甲烷产率,但同时热碱解却使得厌氧消化后污泥部分ARGs(sul1、tetO、tetW、tetX)相对丰度回升.研究表明,延长tHRT有利于taMTPAD工艺削减ARGs和VS,而缩短tHRT则有利于产气.     

新型两相一体厌氧消化反应器处理餐厨垃圾中试研究

针对餐厨垃圾厌氧消化过程易发生挥发性脂肪酸（VFAs）的积累,系统难以稳定高效运行等问题,本研究设计制造了一套中试规模的新型双环嵌套式两相一体厌氧反应器,采用双环嵌套式结构,在反应器内部将内环的产酸阶段与外环的产甲烷阶段分离.反应器共运行184d,初始负荷为60kg/d,运行阶段阶梯式提高负荷,最高达到180kg/d.结果表明,该新型反应器产气与有机固体去除效率高,稳定性好.在稳定运行阶段,有机负荷为3.0kgVS/（m³·d）,容积产气率平均达到1.69m³/（m³·d）,最高为2.72m³/（m³·d）,平均负荷产气率为0.57m³/（kgVS·d）,有机固体去除率达77.2%;由于内外环相分离作用,外环pH值相比内环高约0.3~0.4,外环挥发性脂肪酸（VFAs）和溶解性化学需氧量（sCOD）平均浓度相比内环分别降低25%和23%.双环嵌套式反应器实现了产酸和产甲烷阶段分离的作用,内环产酸相很好地完成了水解和酸化步骤,同时减轻产甲烷相受到酸积累的冲击,对反应器稳定运行起到关键作用.     

Waste activated sludge treatment based on temperature staged and biologically phased anaerobic digestion system

The concept of temperature staged and biological phased （TSBP） was proposed to enhance the performance of waste-activated sludge anaerobic digestion. Semi-continuous experiments were used to investigate the effect of temperature （35 to 70℃） as well as the hydraulic retention time （HRT） （2, 4 and 6 days） on the acidogenic phase. The results showed that the solubilization degree of waste- activated sludge increased from 14.7% to 30.1% with temperature increasing from 35 to 70℃, while the acidification degree was highest at 45℃ （17.6%）, and this was quite different from the temperature impact on hydrolysis. Compared with HRT of 2 and 6 days, 4 days was chosen as the appropriate HRT because of its relatively high solubilization degree （24.6%） and acidification degree （20.1%） at 45℃. The TSBP system combined the acidogenic reactor （45℃, 4 days） with the methanogenic reactor （35℃, 16 days） and the results showed 84.8% and 11.4% higher methane yield and volatile solid reduction, respectively, compared with that of the single-stage anaerobic digestion system with HRT of 20 days at 35℃. Moreover, different microbial morphologies were observed in the acidogenic- and methanogenic-phase reactors, which resulted from the temperature control and HRT adjustment. All the above results indicated that 45℃ was the optimum temperature to inhibit the activity of methanogenic bacteria in the acidogenic phase, and temperature staging and phase separation was thus accomplished. The advantages of the TSBP process were also confirmed by a full-scale waste-activated sludge anaerobic digestion project which was an energy self-sufficient system.     

改性污泥基吸附剂深度处理出水有机物的研究

本研究通过对污泥热解碳化,并负载传统的混凝剂制成改性污泥基吸附剂（Modified Sludge-Based Adsorbents, MSBAs）,用来深度处理超短污泥龄活性污泥工艺出水有机物（COD）.同时,以总有机物（TCOD）和溶解性有机物（SCOD）去除率及处理前后各类有机物组分特征变化为指标,评价MSBAs深度处理出水中有机物的效果及处理后污水中有机物的潜在风险.结果表明,改性后的污泥基吸附剂均有较好的有机物吸附效果,TCOD的去除效率可达到32.6%以上,其中,Fe₂（SO₄）₃改性制备的污泥基吸附剂对有机物的去除能力最大,TCOD最大的去除效率为42.5%,其中,SCOD最大的去除效率为44.1%.有机物特征分析结果显示,4种MSBAs对污水中疏水酸性有机物（HPO-A）有较好的去除效果,且处理后污水中有机物芳香构造化程度增加,稳定性增加,污水消毒副产物的产生可能性大大降低.     

脱硫石膏对稻田CH4释放及其功能微生物种群的影响

为探究脱硫石膏对温室气体CH_4排放及其功能微生物种群的影响,采用静态暗箱/气相色谱法,高通量测序和荧光定量PCR技术,研究了FGDG0(0 t·hm~(-2))、FGDG1(2 t·hm~(-2))、FGDG2(4 t·hm~(-2))、FGDG3(8 t·hm~(-2))、FGDG4(16 t·hm~(-2))这5个施加脱硫石膏处理下CH_4排放特征、稻田细菌群落结构及产甲烷菌和甲烷氧化菌丰度的变化.结果表明施加脱硫石膏后,土壤p H显著提高(P0.05),土壤氧化还原电位、有机质、速效钾含量增加,但处理间无显著差异(P0.05);稻田CH_4平均排放通量随着脱硫石膏用量增加而降低,且FGDG1FGDG2FGDG3FGDG4,较对照分别减少31.56%、57.30%、83.60%、90.66%;与对照相比,FGDG1、FGDG2处理增加了土壤细菌丰富度和多样性,但用量超过4 t·hm~(-2)后,细菌丰富度和多样性逐渐降低;与对照相比,稻田土壤硫酸盐还原菌属相对丰度显著提高6.98%~13.56%,甲烷氧化菌pmo A基因丰度增加0.3%~6.2%,产甲烷菌mrc A基因丰度显著下降2.4%~15.8%,且丰度比(pmo A/mcr A)随着脱硫石膏用量增加而增大;相关性分析表明,CH_4平均排放通量与土壤中硫酸盐还原菌属相对丰度呈显著负相关,与产甲烷菌mcr A基因丰度呈显著正相关,与pmo A/mcr A比呈显著负相关.综上,脱硫石膏能够提高稻田土壤细菌群落多样性,抑制稻田CH_4排放.