好氧堆肥高温期灭活病原菌的效果和影响因素研究

doi:10.13560/j.cnki.biotech.bull.1985.2021-0048

摘要/Abstract

摘要：

畜禽粪便和秸秆等农业有机废弃物中存在大量具有致病风险的人体和动植物病原菌,好氧堆肥高温期对病原菌的消减效果和影响病原菌存活的理化因子需探明。首先检测了市售25种商品有机肥料中病原菌存活的情况;其次考察了好氧堆肥高温阶段病原菌的灭活效果,并探究温度、pH、铵浓度对2种模式病原菌（大肠杆菌与青枯菌）存活的影响。调查的25种商品有机肥中有3个样品检测出存活的潜在病原菌,表明市售商品有机肥仍存在环境风险。好氧堆肥高温期对病原菌的灭活试验表明,高温期后堆体中仅检测出丰度极低的病原菌DNA。室内试验表明随着温度的升高,2种模式病原菌的存活能力减弱,灭活的时间缩短;pH对2种病原菌的灭活效果不同;铵浓度越高,对2种病原菌生长的抑制效果越强。高温期是好氧堆肥过程中病原菌灭活的重要阶段,堆肥高温期可有效消减畜禽粪便堆肥原料中的大部分病原菌,达到相关行业标准的要求。堆体温度、pH和铵浓度是影响病原菌存活的重要因素。

关键词: 好氧堆肥, 高温期, 病原菌, 灭活, 参数

Abstract:

There are large numbers of human and animal-plant pathogens at high risk of disease in the agricultural organic waste such as livestock manure and plant straws. Effects of pathogen inactivation during the thermophilic stage of aerobic composting and key factors affecting the survival of pathogens need to be investigated. Firstly,the pathogen survivals in 25 kinds of commercial organic fertilizers on the market were detected. Secondly,pathogen inactivation during the thermophilic stage of aerobic composting were investigated. Effects of temperature,pH and ammonium content on the survival of two model pathogens（Escherichia coli and Ralstonia solanacearum）were explored. Three of the 25 commercial organic fertilizers contained living potential pathogens,indicating that the organic fertilizers on commercial market still had potential environmental threats. The inactivation testing of pathogens during the thermophilic stage of aerobic composting showed that only pathogen DNA at very low abundance was detected in the composting heaps. The results in the laboratory showed that the survival of two model pathogens decreased with the increase of temperature,and the inactivation time became less. The pH presented different effects on the two pathogens. The higher the ammonium content was,the stronger the inhibitory effect on the growth of the two pathogens was. Thermophilic stage is important to pathogen inactivation during aerobic composting. Thermophilic stage during composting may effectively reduce most pathogens in the raw materials of composting such as livestock and poultry manure,which meet the requirements of relevant industry standards. Temperature,pH and ammonium content are important factors affecting pathogen survival in the composting heaps.

Key words: aerobic composting, thermophilic stage, pathogen, inactivation, parameters

杨天杰, 张令昕, 顾少华, 潘子豪, 江高飞, 王世梅, 韦中, 徐阳春, 沈其荣. 好氧堆肥高温期灭活病原菌的效果和影响因素研究[J]. 生物技术通报, 2021, 37(11): 237-247.

YANG Tian-jie, ZHANG Ling-xin, GU Shao-hua, PAN Zi-hao, JIANG Gao-fei, WANG Shi-mei, WEI Zhong, XU Yang-chun, SHEN Qi-rong. Effects on Pathogen Inactivation at the Thermophilic Stage of Aerobic Composting and Its Impact Factors[J]. Biotechnology Bulletin, 2021, 37(11): 237-247.

图/表 9

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样品编号 Sample No.	生产原料 Raw materials	堆肥工艺 Composting process	堆肥时间 Composting time/d
1	牛粪、秸秆、营养剂Cow manure, straws, nutrient	条垛式堆肥Windrow composting	15-21
2	畜禽粪便、秸秆Livestock manure, straws	条垛式堆肥Windrow composting	15-21
3	牛粪、稻壳、猪粪Cow manure, rice husk, pig manure	槽式堆肥Trough composting	>30
4	牛粪、秸秆Cow manure, straws	条垛式堆肥Windrow composting	>30
5	牛粪、米糠、草木灰Cow manure, rice bran, plant ash	条垛式堆肥Windrow composting	<15
6	牛粪、米糠、发酵菌Cow manure, rice bran, fermented microbe	条垛式堆肥Windrow composting	22-30
7	沼渣、猪粪、鸡粪、菇渣、甜叶菊渣Biogas residues, pig manure, chicken manure, mushroom residues, stevia residues	条垛式堆肥Windrow composting	>30
8	鸡粪、羊粪Chicken manure, sheep manure	静态通气堆肥Static aeration composting	22-30
9	菌菇渣、牛粪、糠醛渣、梨渣Mushroom residues, cow manure, furfural residues, pear residues	条垛式堆肥Windrow composting	22-30
10	木薯渣、牛粪、菌菇渣Cassava residues, cow manure, mushroom residues	条垛式堆肥Windrow composting	>30
11	木薯渣、鸡粪、菌菇渣Cassava residues, chicken manure, mushroom residues	条垛式堆肥Windrow composting	15-21
12	菌菇渣、羊粪、秸秆、猪粪、草木灰Mushroom residues, sheep manure, straws, pig manure, plant ash	条垛式堆肥Windrow composting	>30
13	牛粪、木薯渣、猪粪、氨基酸Cow manure, cassava residue, pig manure, amino acids	条垛式堆肥Windrow composting	15-21
14	牛粪、糠醛渣、蘑菇渣、骨粉Cow manure, furfural residues, mushroom residues, bone meal	条垛式堆肥Windrow composting	>30
15	鸡粪、稻壳、秸秆Chicken manure, rice husk, straws	条垛式堆肥Windrow composting	15-21
16	畜禽粪便、菌渣Livestock manure, bacterial residues	条垛式堆肥Windrow composting	<15
17	鲜鸡粪、垫料鸡粪Fresh chicken manure, litter chicken manure	条垛式堆肥Windrow composting	22-30
18	畜禽粪便、食用菌培养棒Livestock manure, edible fungus culture sticks	条垛式堆肥Windrow composting	20-25

样品编号 Sample No.	生产原料 Raw materials	堆肥工艺 Composting process	堆肥时间 Composting time/d
1	牛粪、秸秆、营养剂Cow manure, straws, nutrient	条垛式堆肥Windrow composting	15-21
2	畜禽粪便、秸秆Livestock manure, straws	条垛式堆肥Windrow composting	15-21
3	牛粪、稻壳、猪粪Cow manure, rice husk, pig manure	槽式堆肥Trough composting	>30
4	牛粪、秸秆Cow manure, straws	条垛式堆肥Windrow composting	>30
5	牛粪、米糠、草木灰Cow manure, rice bran, plant ash	条垛式堆肥Windrow composting	<15
6	牛粪、米糠、发酵菌Cow manure, rice bran, fermented microbe	条垛式堆肥Windrow composting	22-30
7	沼渣、猪粪、鸡粪、菇渣、甜叶菊渣Biogas residues, pig manure, chicken manure, mushroom residues, stevia residues	条垛式堆肥Windrow composting	>30
8	鸡粪、羊粪Chicken manure, sheep manure	静态通气堆肥Static aeration composting	22-30
9	菌菇渣、牛粪、糠醛渣、梨渣Mushroom residues, cow manure, furfural residues, pear residues	条垛式堆肥Windrow composting	22-30
10	木薯渣、牛粪、菌菇渣Cassava residues, cow manure, mushroom residues	条垛式堆肥Windrow composting	>30
11	木薯渣、鸡粪、菌菇渣Cassava residues, chicken manure, mushroom residues	条垛式堆肥Windrow composting	15-21
12	菌菇渣、羊粪、秸秆、猪粪、草木灰Mushroom residues, sheep manure, straws, pig manure, plant ash	条垛式堆肥Windrow composting	>30
13	牛粪、木薯渣、猪粪、氨基酸Cow manure, cassava residue, pig manure, amino acids	条垛式堆肥Windrow composting	15-21
14	牛粪、糠醛渣、蘑菇渣、骨粉Cow manure, furfural residues, mushroom residues, bone meal	条垛式堆肥Windrow composting	>30
15	鸡粪、稻壳、秸秆Chicken manure, rice husk, straws	条垛式堆肥Windrow composting	15-21
16	畜禽粪便、菌渣Livestock manure, bacterial residues	条垛式堆肥Windrow composting	<15
17	鲜鸡粪、垫料鸡粪Fresh chicken manure, litter chicken manure	条垛式堆肥Windrow composting	22-30
18	畜禽粪便、食用菌培养棒Livestock manure, edible fungus culture sticks	条垛式堆肥Windrow composting	20-25

病原菌Pathogen	堆体Pile	堆肥时间Composting time/d	OTU数量OTU abundance
白葡萄球菌 Saccharopolyspora rectivirgula	A	0	176.0±83.44 a
	A	8	1.33±1.15 b
	B	0	27.0±6.0 a
	B	8	1.0±0 b
鹿角球菌 Vagococcus lutrae	A	0	4.5±2.12 a
	A	8	1.33±1.53 a
	B	0	75.33±3.21 a
	B	8	0 b
Paenalcaligenes hominis	A	0	0
	A	8	0
	B	0	22.67±8.08 a
	B	8	3.67±2.31 b
鲍曼不动杆菌 Acinetobacter baumannii	A	0	26±12.73 a
	A	8	0 b
	B	0	6.0±3.61 a
	B	8	0 b
深红沙雷氏菌 Serratia rubidaea	A	0	29.5±6.36 a
	A	8	0 b
	B	0	2.0±1.0 a
	B	8	0.33±0.58 a
Chitinophaga terrae	A	0	23.5±28.99 a
	A	8	0 a
	B	0	0.67±0.58 a
	B	8	0 a
大肠埃希-志贺氏菌 Escherichia-Shigella	A	0	15.5±0.71 a
	A	8	0 b
	B	0	3.0±2.65 a
	B	8	0 a
纤维化纤维微细菌 Cellulosimicrobium cellulans	A	0	6.0±5.65 a
	A	8	0.33±0.58 a
	B	0	5.33±2.31 a
	B	8	0 b
嗜麦芽窄食单胞菌 Stenotrophomonas maltophilia	A	0	13.5±3.54 a
	A	8	0 b
	B	0	0.33±0.58 a
	B	8	0 a
抗热分枝杆菌 Mycobacterium thermoresistibile	A	0	2.0±0 a
	A	8	2.0±3.46 a
	B	0	0 a
	B	8	0.33±0.58 a
Pandoraea pnomenusa	A	0	4.0±1.41 a
	A	8	0 b
	B	0	0.67±0.58 a
	B	8	0 a
格氏李斯特菌 Listeria grayi	A	0	2.5±0.71 a
	A	8	0 b
	B	0	0.33±0.58 a
	B	8	0 a

病原菌Pathogen	堆体Pile	堆肥时间Composting time/d	OTU数量OTU abundance
白葡萄球菌 Saccharopolyspora rectivirgula	A	0	176.0±83.44 a
	A	8	1.33±1.15 b
	B	0	27.0±6.0 a
	B	8	1.0±0 b
鹿角球菌 Vagococcus lutrae	A	0	4.5±2.12 a
	A	8	1.33±1.53 a
	B	0	75.33±3.21 a
	B	8	0 b
Paenalcaligenes hominis	A	0	0
	A	8	0
	B	0	22.67±8.08 a
	B	8	3.67±2.31 b
鲍曼不动杆菌 Acinetobacter baumannii	A	0	26±12.73 a
	A	8	0 b
	B	0	6.0±3.61 a
	B	8	0 b
深红沙雷氏菌 Serratia rubidaea	A	0	29.5±6.36 a
	A	8	0 b
	B	0	2.0±1.0 a
	B	8	0.33±0.58 a
Chitinophaga terrae	A	0	23.5±28.99 a
	A	8	0 a
	B	0	0.67±0.58 a
	B	8	0 a
大肠埃希-志贺氏菌 Escherichia-Shigella	A	0	15.5±0.71 a
	A	8	0 b
	B	0	3.0±2.65 a
	B	8	0 a
纤维化纤维微细菌 Cellulosimicrobium cellulans	A	0	6.0±5.65 a
	A	8	0.33±0.58 a
	B	0	5.33±2.31 a
	B	8	0 b
嗜麦芽窄食单胞菌 Stenotrophomonas maltophilia	A	0	13.5±3.54 a
	A	8	0 b
	B	0	0.33±0.58 a
	B	8	0 a
抗热分枝杆菌 Mycobacterium thermoresistibile	A	0	2.0±0 a
	A	8	2.0±3.46 a
	B	0	0 a
	B	8	0.33±0.58 a
Pandoraea pnomenusa	A	0	4.0±1.41 a
	A	8	0 b
	B	0	0.67±0.58 a
	B	8	0 a
格氏李斯特菌 Listeria grayi	A	0	2.5±0.71 a
	A	8	0 b
	B	0	0.33±0.58 a
	B	8	0 a

堆体理化性质 Physico-chemical properties	病原菌数量Pathogen abundance
堆体理化性质 Physico-chemical properties	自由度 Df	F值 F value	P值 P value
温度Temperature/℃	1	86.21	< 0.001
pH	1	23.63	< 0.01
铵态氮含量 Ammonium content/（mg·g^-1）	1	0.12	0.74
残差Residuals	7