秸秆类型及配比变化对污泥厌氧消化中微生物群落的影响

doi:10.16085/j.issn.1000-6613.2019-0777

摘要/Abstract

摘要：

为探讨秸秆类型及配比变化对污泥厌氧消化体系中微生物群落结构的影响，采用16S rRNA高通量测序技术，对污泥-秸秆联合厌氧消化体系中微生物群落进行分析。结果表明：玉米秸秆的添加对体系中pH和挥发性脂肪酸（VFAs）的影响较大，尤其是乙酸。而小麦和水稻秸秆的添加对碱度的影响较大，但当配比增加时体系中的VFAs和乙酸浓度也会增加，特别是1∶1.5（挥发性固体质量比）。通过对微生物群落结构分析发现，秸秆类型及配比的变化能显著提高厌氧体系中水解菌和酸化菌的相对丰度（p＜0.001），如深古菌门Bathyarchaeota、未识别的_c_深古菌门（norank_p_Bathyarchaeota）、拟杆菌门（Bacteroidetes）、阴沟单胞菌门（Cloacimonetes）、同力菌门（Synergistetes）、未分类的_p_阴沟单胞菌门（unclassified_p_Cloacimonetes）、未识别的_c_拟杆菌门_vadinHA17（norank_c_Bacteroidetes_vadinHA17）、（Christensenellaceae_R-7_group）、未识别的_f_紫单胞菌科（norank_f_ Porphyromonadaceae）、未分类的_f_瘤胃菌科（unclassified_f_Ruminococcaceae）、拟杆菌属（Bacteroides）、丙酸杆菌属（Prolixibacter）、长绳菌属（Longilinea）、纤绳菌属（Leptolinea）等菌群；但对体系中乙酸型产甲烷菌（Methanosaeta）和氢型产甲烷菌（甲烷螺菌属Methanospirillum、甲烷杆菌属Methanobacterium和甲烷短杆菌属Methanobrevibacter）的生长产生抑制作用。同时也发现秸秆的添加对甲基型产甲烷菌（Methanomassiliicoccus）的生长具有显著的促进作用（p＜0.001），从而影响厌氧体系的产甲烷特性。

关键词: 秸秆类型, 配比, 微生物群落, 联合厌氧消化, 污泥

Abstract:

In order to investigate the effects of straw type and ratio change on microbial community structure in sludge anaerobic digestion system, 16S rRNA high-throughput sequencing technology was used to analyze the microbial community in sludge-straw anaerobic co-digestion system. The results showed that addition of corn straw had a great influence on the pH and volatile fatty acids (VFAs) in the system, especially on acetic acid. Addition of wheat and rice straw had a greater effect on alkalinity, but the VFAs and acetic acid concentrations in the system increased when the ratio increased, especially 1∶1.5 (mass ratio of volatile solids). Through the analysis of microbial community structure, it was found that the change of straw ratio and types could significantly increase the relative abundance of hydrolysis bacteria and acidification bacteria (p＜0.001) in anaerobic systems, for instance, Bathyarchaeota, norank_p_Bathyarchaeota (archaea), Bacteroidetes, Cloacimonetes, Synergistetes, unclassified_p_ Cloacimonetes, norank_c_Bacteroidetes_vadinHA17, Christensenellaceae_R-7_group, norank_f_Porphyromonadaceae, unclassified_f_Ruminococcaceae, Bacteroides, Prolixibacter, Longilinea and Leptolinea (bacteria), but the growth of methanogens (Methanosaeta) and hydrogen methanogenic bacteria (Methanospirillum, Methanobacterium and Methanobrevibacter) could be inhibited. It was also found that the addition of straw significantly promoted the growth of methylotrophic methanogens (Methanomassiliicoccus) (p＜0.001), which affected the methanogenic properties of anaerobic systems.

Key words: straw type, ratio, microbial community, anaerobic co-digestion, sludge

中图分类号:

X172

唐涛涛,李江,杨爱江,杨钊,向福亮,袁华宇. 秸秆类型及配比变化对污泥厌氧消化中微生物群落的影响[J]. 化工进展, 2020, 39(2): 667-678.

Taotao TANG,Jiang LI,Aijinag YANG,Zhao YANG,Fuliang XIANG,Huayu YUAN. Effects of straw type and ratio change on microbial community in anaerobic digestion of sludge[J]. Chemical Industry and Engineering Progress, 2020, 39(2): 667-678.

图/表 10

表1 污泥与秸秆基本性质

样品	pH	化学需氧量COD/mg·L^-1	含水率/%	$V S T S$ /%
污泥	7.2±0.25	24257.07±2313.10	96.91±0.54	51.57±7.47
玉米	—	—	19.92±6.17	92.62±5.83
小麦	—	—	15.41±0.42	88.34±0.56
水稻	—	—	15.76±0.49	82.26±0.88

表1 污泥与秸秆基本性质

样品	pH	化学需氧量COD/mg·L^-1	含水率/%	$V S T S$ /%
污泥	7.2±0.25	24257.07±2313.10	96.91±0.54	51.57±7.47
玉米	—	—	19.92±6.17	92.62±5.83
小麦	—	—	15.41±0.42	88.34±0.56
水稻	—	—	15.76±0.49	82.26±0.88

表2 秸秆对消化污泥特性的影响

试验号	累积沼气产量/L	pH	$N H 4 + - N$ /mg·L^-1	碱度/mg·L^-1	VFAs/mg·L^-1	VFAs/ALK
CK	52.43±1.67	7.31±0.16	324.43±13.28	1304.54±93.35	181.27±21.04	0.14±0.02
C1	106.13±5.80	7.62±0.26	478.33±27.05	1860.25±59.02	203.50±20.45	0.11±0.01
C2	163.02±3.99	6.94±0.23	293.42±7.00	1445.59±289.01	261.76±40.78	0.19±0.04
C3	223.98±8.63	6.89±0.24	262.89±9.54	1367.77±305.71	295.66±31.38	0.22±0.04
W1	93.82±3.71	7.76±0.13	449.94±17.56	1710.85±124.10	196.85±18.81	0.12±0.01
W2	150.88±4.55	7.39±0.18	442.44±12.96	1669.67±125.13	207.02±24.85	0.12±0.02
W3	211.5±5.17	7.51±0.20	536.0±38.96	2016.29±134.44	225.68±15.33	0.11±0.01
R1	106.91±1.86	7.44±0.18	390.19±15.52	1707.19±69.69	202.21±22.55	0.12±0.02
R2	149.99 ± 21.98	7.37±0.19	481.13±25.59	1869.41±77.10	211.83±21.05	0.11±0.01
R3	211.68 ± 11.29	7.42±0.24	542.52±25.96	2044.33±142.87	237.29±21.79	0.12±0.01
试验号	乙酸/mg·L^-1	丙酸/mg·L^-1	丁酸/mg·L^-1	异丁酸/mg·L^-1	戊酸/mg·L^-1
CK	57.24±19.69	16.17±3.80	21.95±1.17	30.19±2.00	26.55±6.82
C1	80.79±20.06	17.80±2.08	23.87±4.44	29.87±1.04	23.11±0.96
C2	119.89±35.62	19.78±4.94	23.35±3.31	34.88±3.95	23.80±3.28
C3	156.26±26.44	16.64±3.82	22.46±1.82	34.23±4.45	23.40±1.74
W1	68.04±16.06	21.89±2.81	25.43±5.60	28.86±0.91	23.18±1.03
W2	77.85±15.72	22.61±7.06	23.76±2.38	29.03±0.94	24.93±1.61
W3	99.54±11.52	21.22±5.57	23.59±1.44	30.09±1.12	24.50±1.82
R1	70.37±20.28	21.61±3.38	23.76±3.83	32.37±6.62	24.04±2.68
R2	83.33±18.35	20.83±2.96	25.21±4.13	29.66±1.55	23.40±1.89
R3	107.55±19.54	20.60±6.84	24.10±2.82	31.07±4.41	25.97±8.09

表2 秸秆对消化污泥特性的影响

试验号	累积沼气产量/L	pH	$N H 4 + - N$ /mg·L^-1	碱度/mg·L^-1	VFAs/mg·L^-1	VFAs/ALK
CK	52.43±1.67	7.31±0.16	324.43±13.28	1304.54±93.35	181.27±21.04	0.14±0.02
C1	106.13±5.80	7.62±0.26	478.33±27.05	1860.25±59.02	203.50±20.45	0.11±0.01
C2	163.02±3.99	6.94±0.23	293.42±7.00	1445.59±289.01	261.76±40.78	0.19±0.04
C3	223.98±8.63	6.89±0.24	262.89±9.54	1367.77±305.71	295.66±31.38	0.22±0.04
W1	93.82±3.71	7.76±0.13	449.94±17.56	1710.85±124.10	196.85±18.81	0.12±0.01
W2	150.88±4.55	7.39±0.18	442.44±12.96	1669.67±125.13	207.02±24.85	0.12±0.02
W3	211.5±5.17	7.51±0.20	536.0±38.96	2016.29±134.44	225.68±15.33	0.11±0.01
R1	106.91±1.86	7.44±0.18	390.19±15.52	1707.19±69.69	202.21±22.55	0.12±0.02
R2	149.99 ± 21.98	7.37±0.19	481.13±25.59	1869.41±77.10	211.83±21.05	0.11±0.01
R3	211.68 ± 11.29	7.42±0.24	542.52±25.96	2044.33±142.87	237.29±21.79	0.12±0.01
试验号	乙酸/mg·L^-1	丙酸/mg·L^-1	丁酸/mg·L^-1	异丁酸/mg·L^-1	戊酸/mg·L^-1
CK	57.24±19.69	16.17±3.80	21.95±1.17	30.19±2.00	26.55±6.82
C1	80.79±20.06	17.80±2.08	23.87±4.44	29.87±1.04	23.11±0.96
C2	119.89±35.62	19.78±4.94	23.35±3.31	34.88±3.95	23.80±3.28
C3	156.26±26.44	16.64±3.82	22.46±1.82	34.23±4.45	23.40±1.74
W1	68.04±16.06	21.89±2.81	25.43±5.60	28.86±0.91	23.18±1.03
W2	77.85±15.72	22.61±7.06	23.76±2.38	29.03±0.94	24.93±1.61
W3	99.54±11.52	21.22±5.57	23.59±1.44	30.09±1.12	24.50±1.82
R1	70.37±20.28	21.61±3.38	23.76±3.83	32.37±6.62	24.04±2.68
R2	83.33±18.35	20.83±2.96	25.21±4.13	29.66±1.55	23.40±1.89
R3	107.55±19.54	20.60±6.84	24.10±2.82	31.07±4.41	25.97±8.09

表3 不同实验组中细菌群落的丰度和多样性

样品名称	OTU	Chao	ACE	Shannon	Simpson	Coverage
CK	1301	1537.99	1515.78	5.28	0.017	0.9920
C1	1613	1805.21	1811.09	5.39	0.023	0.9925
C2	1595	1816.65	1785.31	4.98	0.043	0.9950
C3	1349	1622.81	1631.66	5.15	0.025	0.9879
W1	1572	1800.11	1781.41	5.45	0.016	0.9918
W2	1628	1835.45	1833.47	5.44	0.022	0.9923
W3	1346	1662.75	1658.87	4.90	0.030	0.9906
R1	1608	1864.64	1819.15	5.54	0.021	0.9923
R2	1627	1870.63	1849.14	5.37	0.019	0.9927
R3	1642	1805.18	1816.58	5.35	0.025	0.9942

表4 不同实验组中古菌群落的丰度和多样性

样品名称	OTU	Chao	ACE	Shannon	Simpson	Coverage
CK	59	60.11	60.83	1.82	0.285	0.9999
C1	62	69.88	78.69	1.96	0.260	0.9997
C2	48	50.00	50.68	1.65	0.366	0.9999
C3	55	63.20	62.93	2.01	0.227	0.9997
W1	110	119.50	120.98	2.01	0.237	0.9995
W2	60	65.58	72.47	1.93	0.261	0.9997
W3	60	73.00	67.90	1.65	0.376	0.9998
R1	82	125.17	123.88	2.14	0.191	0.9995
R2	92	99.56	104.77	2.35	0.147	0.9997
R3	78	125.00	139.07	1.80	0.296	0.9993

图1 OTU的Venn图

图2 细菌和古菌在门和属水平下的分类Bacteroidetes (拟杆菌门）；Proteobacteria（变形菌门）；Chloroflexi（绿弯菌门）；Firmicutes（厚壁菌门）；Actinobacteria（放线菌门）；Acidobacteria（酸杆菌门）；Verrucomicrobia（疣微菌门）；Nitrospirae（硝化螺旋菌门）；Caldiserica（嗜热丝菌门）；Planctomycetes（浮霉菌门）；Fibrobacteres（纤维秆菌门）；Euryarchaeota（广古菌门）（门分类）；Bacteroides（拟杆菌属）；Nitrospira（硝化螺旋菌属）；Woodsholea（木洞菌属）；Syntrophobacter（互营杆菌属）；Syntrophus（互营菌属）；Smithella（史密斯氏菌属）；Roseiflexus（花弯菌属）； norank_f_Porphyromonadaceae（未识别的_f_紫单胞菌科）；norank_f_Anaerolineaceae（未识别的_f_厌氧绳菌科）；norank_f_Saprospiraceae（未识别的_f_腐螺旋菌科）；norank_k_Acidobacteria（未识别的_k_酸杆菌门）；norank_f_Xanthomonadaceae（未识别的_f_黄单胞菌科）；unclassified_f_ Ruminococcaceae（未分类的_f_瘤胃菌科）；unclassified_p_Chloroflexi（未分类的_p_绿弯菌门）；unclassified_f_Anaerolineaceae（未分类的_f_厌氧绳菌科）；norank_o_Acidimicrobiales（未识别的_o_酸微菌目）；unclassified_f_Peptostreptococcaceae（未分类的_f_消化链球菌科）；Methanosaeta （鬃毛甲烷菌属）；Methanobacterium（甲烷杆菌属）；Methanospirillum（甲烷螺菌属）；Methanobrevibacter（甲烷短杆菌属）（属分类）

图3 各样本间的分布热图Leptolinea（纤绳菌属）；Methanomethylovorans（甲烷食甲基菌属）；Methanosarcina（甲烷八叠球菌属）；Methanosphaera（甲烷球菌属）；Methanolinea（甲烷绳菌属）；Methanoculleus（甲烷囊菌属）；Methanocorpusculum（甲烷粒菌属）（属分类）

图4 细菌和古菌在属分类上的主成分分析

图5 各样本间的RDA分析

表5 环境因子的VIF值

环境因子	VIF
环境因子	筛选前	筛选后
pH	51.81	7.15
氨氮（ $N H 4 + - N$ ）	3631.04	4.34
乙酸	248838.3	3.96
丙酸	1474.49	2.42
丁酸	711.34	2.88
异丁酸	4888.83	4.67

表5 环境因子的VIF值

环境因子	VIF
环境因子	筛选前	筛选后
pH	51.81	7.15
氨氮（ $N H 4 + - N$ ）	3631.04	4.34
乙酸	248838.3	3.96
丙酸	1474.49	2.42
丁酸	711.34	2.88
异丁酸	4888.83	4.67

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