中间产物对干式厌氧消化影响的研究
摘要
干式厌氧消化较湿式厌氧消化有诸多优点,具有更广阔的商业前景,但是也存在许多问题亟待解决：例如传质问题,启动时间长,产气不稳定等。系统的稳定性取决于参与消化过程的细菌活性,利用荧光原位杂交(FISH)技术检测不同条件下中温干式厌氧消化过程中的特定微生物,探讨干式厌氧消化过程中间产物对产甲烷的影响,分析挥发性脂肪酸(VFA)和相应菌群之间的关系,同时研究该过程中乙酸、丙酸、丁酸对硫酸盐还原菌(SRB)的影响,主要研究结果如下：
(1)在消化反应器中添加乙酸,每两天添加一次,共8次,发现添加量为10.5g时,能显著增加反应器中产甲烷菌活性,产生更多的沼气,缩短消化反应的启动时间。反应器内食乙酸产甲烷菌的平均丰度高于食氢产甲烷菌,因为在干式厌氧消化前期,食氢产甲烷菌丰度处于较高水平,随着消化反应的进行,食乙酸产甲烷菌逐渐处于主导地位。对过程中乙酸浓度与产甲烷菌丰度进行关联,得到：乙酸浓度在0-25g·L-1范围内与食氢产甲烷菌,食乙酸产甲烷菌丰度均呈负相关,相关系数R2分别为0.8658,0.8074。
(2)添加丙酸,每两天添加一次,共6次,添加量为4g时,较其他添加量时的产沼气量大,更能促进产甲烷菌的生长。丙酸浓度在0-100g·L-1范围内与食丙酸产氢产乙酸菌丰度呈负相关,相关系数R2=0.8352,丙酸的浓度在0-30g·L-1范围内的食丙酸产氢产乙酸菌丰度处于较高水平。
(3)添加丁酸,每两天添加一次,共6次,添加量为4.8g时,较其他添加量可显著增加反应器中产甲烷菌的活性。丁酸浓度在0-2.4g.L-1范围内与食丁酸产氢产乙酸菌丰度呈正相关,相关系数R2=0.7229；乙酸浓度在0-30g·L-1与食丁酸产氢产乙酸菌丰度呈正相关,相关系数R2=0.8100,可推知,乙酸浓度与参与该反应微生物丰度的关联性更好。
(4)在中温干式厌氧消化过程中,分别添加最佳量的乙酸,丙酸,丁酸,并以空白组作对照,研究VFAs对SRB还原速率的贡献,结果显示：丙酸>丁酸>乙酸。利用FISH技术监测添加不同VFAs的反应器内SRB与产甲烷菌(MB)种群的演变,研究表明,各反应器中SRB与MB的活性均为：添加丙酸>添加乙酸＞添加丁酸＞对照,得出添加一定量的丙酸更有利于激活SRB的活性,促进厌氧消化反应器中丙酸的转化,增强SRB与MB种群间的协同作用,提高厌氧反应器抵御“酸败”的能力。
Dry anaerobic digestion has great commercial prospects, which has much more advantages than wet anaerobic digestion. However, there are still many problems to be solved:such as mass transfer problem, startup time, stable gas, etc. It's worth to note that the stability of the system depends on the activity of bacteria participating in the digestive process. In this work, the specific microorganisms in anaerobic digestion in different conditions were detectde using fluorescence in situ hybridization (FISH). The effect of intermediate to produce methane in dry anaerobic digestion process was explored. Meanwhile, the relationship of VFAs and the corresponding microorganisms was analyzed. In addition, the influence of VFAs, which include acetic acid, propionic acid and butyric acid, to the sulfate reducing bacteria in anaerobic digestion was also investigated. The main results of this study were as follows:
(1) The optimal amount of acetic acid added was10.5g every other day for eight times till to the stability stage. This could avoid inhibiting the active of microbial community, produce more biogas, and short the startup time of anaerobic digestion. The average population of the H2-utilizing methanogen was more than the acetate-utilizing methanogen. At the start-up of the experiment the population of H2-utilizing methanogen increased to the maximum and decreased sharply. At stable stage the level of H2-utilizing methanogens was very low, in contrast, the population of acetate-utilizing methanogen increased. The relationship of the content of acetic acid with the population of H2-utilizing methanogen and acetate-utilizing methanogen were all negative linear with the coefficient of0.8658and0.8074, respectively, when the content of acetic acid was0-25g·L-1.
(2) The optimal amount of propionic acid added was10.5g every other day for six times. Under such condition, biogas production quantity was larger than other reactors, which could promote the growth of methanogens. The relationship of the content of propionic acid and the population of Syntrophic propionate-oxidizing bacteria were negative linear with the coefficient of0.8352, when the content of propionic acid was0-100g·L-1. Meanwhile, concentration of propionic acid in0to30g·L-1was accompanied by the higher level of Syntrophic propionate-oxidizing bacteria.
(3) The optimal amount of butyric acid added was4.8g every other day for six times. This could increase the activity of methane bacteria. The relationship of butyric acid and Syntrophic butyrate-oxidizing bacteria were positive linear with the coefficient of0.7229, when the content of butyric acid was0-2.4g·L-1. The relationship of the content of acetic acid and the population of Syntrophic butyrate-oxidizing bacteria were positive linear with the coefficient of0.8100, when the content of acetic acid was0-30g·L-1. It indicated that the correlation between concentration of acetic acid and the microbial involved in the response was better.
(4) Added the optimal amount of acetic acid, propionic acid, butyric acid, the contribution of VFAs to reduction rate of SRB was investigated, results showed: acrylic acid> butyric acid> acetic acid. The evolution of populations of SRB and MB was monitored by using FISH technology in the reactor by adding different VFAs, studies showed that the activity of MB and SRB in the reactors was additional butyric acid> additional acetic acid> additional propionic acid> controls. Adding a certain amount of propionic acid was more advantageous to activate the activity of SRB, promote the transformation of propionic acid, enhance synergy of SRB and MB, improve the ability of against the rancidity.
(1)在消化反应器中添加乙酸,每两天添加一次,共8次,发现添加量为10.5g时,能显著增加反应器中产甲烷菌活性,产生更多的沼气,缩短消化反应的启动时间。反应器内食乙酸产甲烷菌的平均丰度高于食氢产甲烷菌,因为在干式厌氧消化前期,食氢产甲烷菌丰度处于较高水平,随着消化反应的进行,食乙酸产甲烷菌逐渐处于主导地位。对过程中乙酸浓度与产甲烷菌丰度进行关联,得到：乙酸浓度在0-25g·L-1范围内与食氢产甲烷菌,食乙酸产甲烷菌丰度均呈负相关,相关系数R2分别为0.8658,0.8074。
(2)添加丙酸,每两天添加一次,共6次,添加量为4g时,较其他添加量时的产沼气量大,更能促进产甲烷菌的生长。丙酸浓度在0-100g·L-1范围内与食丙酸产氢产乙酸菌丰度呈负相关,相关系数R2=0.8352,丙酸的浓度在0-30g·L-1范围内的食丙酸产氢产乙酸菌丰度处于较高水平。
(3)添加丁酸,每两天添加一次,共6次,添加量为4.8g时,较其他添加量可显著增加反应器中产甲烷菌的活性。丁酸浓度在0-2.4g.L-1范围内与食丁酸产氢产乙酸菌丰度呈正相关,相关系数R2=0.7229；乙酸浓度在0-30g·L-1与食丁酸产氢产乙酸菌丰度呈正相关,相关系数R2=0.8100,可推知,乙酸浓度与参与该反应微生物丰度的关联性更好。
(4)在中温干式厌氧消化过程中,分别添加最佳量的乙酸,丙酸,丁酸,并以空白组作对照,研究VFAs对SRB还原速率的贡献,结果显示：丙酸>丁酸>乙酸。利用FISH技术监测添加不同VFAs的反应器内SRB与产甲烷菌(MB)种群的演变,研究表明,各反应器中SRB与MB的活性均为：添加丙酸>添加乙酸＞添加丁酸＞对照,得出添加一定量的丙酸更有利于激活SRB的活性,促进厌氧消化反应器中丙酸的转化,增强SRB与MB种群间的协同作用,提高厌氧反应器抵御“酸败”的能力。
Dry anaerobic digestion has great commercial prospects, which has much more advantages than wet anaerobic digestion. However, there are still many problems to be solved:such as mass transfer problem, startup time, stable gas, etc. It's worth to note that the stability of the system depends on the activity of bacteria participating in the digestive process. In this work, the specific microorganisms in anaerobic digestion in different conditions were detectde using fluorescence in situ hybridization (FISH). The effect of intermediate to produce methane in dry anaerobic digestion process was explored. Meanwhile, the relationship of VFAs and the corresponding microorganisms was analyzed. In addition, the influence of VFAs, which include acetic acid, propionic acid and butyric acid, to the sulfate reducing bacteria in anaerobic digestion was also investigated. The main results of this study were as follows:
(1) The optimal amount of acetic acid added was10.5g every other day for eight times till to the stability stage. This could avoid inhibiting the active of microbial community, produce more biogas, and short the startup time of anaerobic digestion. The average population of the H2-utilizing methanogen was more than the acetate-utilizing methanogen. At the start-up of the experiment the population of H2-utilizing methanogen increased to the maximum and decreased sharply. At stable stage the level of H2-utilizing methanogens was very low, in contrast, the population of acetate-utilizing methanogen increased. The relationship of the content of acetic acid with the population of H2-utilizing methanogen and acetate-utilizing methanogen were all negative linear with the coefficient of0.8658and0.8074, respectively, when the content of acetic acid was0-25g·L-1.
(2) The optimal amount of propionic acid added was10.5g every other day for six times. Under such condition, biogas production quantity was larger than other reactors, which could promote the growth of methanogens. The relationship of the content of propionic acid and the population of Syntrophic propionate-oxidizing bacteria were negative linear with the coefficient of0.8352, when the content of propionic acid was0-100g·L-1. Meanwhile, concentration of propionic acid in0to30g·L-1was accompanied by the higher level of Syntrophic propionate-oxidizing bacteria.
(3) The optimal amount of butyric acid added was4.8g every other day for six times. This could increase the activity of methane bacteria. The relationship of butyric acid and Syntrophic butyrate-oxidizing bacteria were positive linear with the coefficient of0.7229, when the content of butyric acid was0-2.4g·L-1. The relationship of the content of acetic acid and the population of Syntrophic butyrate-oxidizing bacteria were positive linear with the coefficient of0.8100, when the content of acetic acid was0-30g·L-1. It indicated that the correlation between concentration of acetic acid and the microbial involved in the response was better.
(4) Added the optimal amount of acetic acid, propionic acid, butyric acid, the contribution of VFAs to reduction rate of SRB was investigated, results showed: acrylic acid> butyric acid> acetic acid. The evolution of populations of SRB and MB was monitored by using FISH technology in the reactor by adding different VFAs, studies showed that the activity of MB and SRB in the reactors was additional butyric acid> additional acetic acid> additional propionic acid> controls. Adding a certain amount of propionic acid was more advantageous to activate the activity of SRB, promote the transformation of propionic acid, enhance synergy of SRB and MB, improve the ability of against the rancidity.
引文
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