除臭菌的筛选及其在堆肥中的应用效果

doi:10.3864/j.issn.0578-1752.2024.13.010

摘要/Abstract

摘要：

【目的】筛选新型除臭菌，解决以鸡粪为主要原料的有机肥厂的臭气排放问题，为好氧堆肥生物除臭技术提供理论依据和支撑。【方法】以鸡粪为筛菌样品，利用定性初筛、驯化富集、分离纯化、定量复筛、拮抗试验以及耐药性试验筛选能够满足多重目标的除臭菌，并通过形态观察与16S rDNA测序鉴定菌株。将筛选获得的除臭菌株制成复合菌剂BH并应用于鸡粪堆肥，共设置3个处理，分别为对照CK（原料中不添加菌剂）、DT1（原料中添加1%除臭菌剂BH）、DT2（原料中添加1%的市售商品除臭菌剂）。在堆肥不同时期，分析测定堆体温度、pH、电导率（EC）、庆大霉素（GM）含量以及NH₃排放量等指标，探讨耐药性除臭菌剂BH对堆肥过程的影响。【结果】初步筛选分离获得15株细菌，命名为BH1—BH15，未筛选到真菌；经复筛发现，菌株BH2、BH5、BH9、BH11、BH12和BH15对NH₃的去除率较高，分别为48.8%、49.4%、45.8%、48.3%、51.0%和51.8%，且各菌株间无拮抗作用。对除臭菌株耐药性研究发现，菌株BH11和BH12对庆大霉素具有较强的耐受性；经形态学特征与分子生物学鉴定，菌株BH11为Bordetella sp.，BH12为Weeksella massiliensis。将菌株BH11和BH12制成复合菌剂BH，其除臭效果显著优于单一菌剂，NH₃去除率高达65.8%。堆肥试验中，各处理高温期均持续9 d以上，实现了鸡粪的无害化处理；堆肥结束时各处理的pH稳定在8.4左右，处于5.5—8.5之间，EC在2.73—3.43 mS·cm^-1之间，低于4 mS·cm^-1，表明物料可用作植物肥料且符合有机肥标准。研究发现，DT2处理的GM降解效率显著高于CK处理，表明商品菌剂能够促进庆大霉素降解。此外，堆肥过程中NH₃的排放主要发生在堆肥升温期与高温期，且高温期的排放量高于升温期；与CK处理相比，菌剂BH显著抑制升温期NH₃的排放，且除臭效果优于商品菌剂；而进入高温期后，商品菌剂除臭效果优于菌剂BH。【结论】筛选获得2株耐药性除臭菌，对NH₃的去除率分别为48.3%（BH11）和51.0%（BH12），制成的复合菌剂BH对NH₃的去除率高达65.8%。将菌剂BH应用于鸡粪堆肥，可以有效减少堆肥过程中NH₃的排放，且各指标均符合堆肥腐熟标准。

关键词: 除臭菌, 耐药性, 筛选, 复合菌剂, 鸡粪, 堆肥

Abstract:

【Objective】 The aim of this study was to screen new deodorizing bacteria for solving the problem of odor emission from organic fertilizer plants with chicken manure as the main raw material, and providing the theoretical basis and support for aerobic composting biological deodorization technology.【Method】 The chicken manure was used as the sample to screen deodorizing bacteria to meet multiple goals by qualitative preliminary screening, domestication and enrichment, separation and purification, quantitative re-screening, antagonism test and drug-resistance test. The strains were identified by morphological observation and 16S rDNA sequence, and the compound bacteria agent BH was prepared and applied to chicken manure composting. Three treatments were set up, including raw materials without biological agents (CK), raw materials mixed with 1% biological deodorizing agent BH (DT1), and raw materials mixed with 1% purchased biological deodorizing agent (DT2). Some indexes were detected, including temperature, pH, electroconductibility (EC), gentamicin (GM) content and NH₃ emission, to explore the effect of BH during composting.【Result】 15 strains of bacteria were screened and isolated, named as BH1-BH15, but no fungi were screened. After rescreening, it was found that strains BH2, BH5, BH9, BH11, BH12 and BH15 had higher removal rates of NH₃, which were 48.8%, 49.4%, 45.8%, 48.3%, 51.0% and 51.8%, respectively, and there was no antagonism among the strains. The study on the drug-resistance of deodorizing strains showed that strains BH11 and BH12 had strong resistance to gentamicin. BH11 was identified as Bordetella sp. and BH12 was identified as Weeksella massiliensis by morphology and molecular biology. Then strain BH11 and BH12 were made into compound bacterial agent BH, and its deodorizing effect was significantly better than that of single bacterial agent, with the removal rate of NH₃ was 65.8%. During composting, the thermophilic phase of each treatment lasted for more than 9 days, and the harmless treatment of chicken manure was achieved. At the end of composting, the pH value of each treatment was stable at 8.40, between 5.5 and 8.5, and the EC was between 2.73 and 3.43 mS·cm^-1, lower than 4 mS·cm^-1, indicated that the material could be used as plant fertilizer and met the organic fertilizer standards. It was found that the GM degradation efficiency of DT2 treatment was significantly higher than that of CK treatment, indicated the commercial bacterial agent could promote the degradation of gentamicin. In addition, the emission of NH₃ during composting mainly occurred in mesophilic phase and thermophilic phase, and the emission in thermophilic phase was higher than that in mesophilic phase. Compared with CK treatment, the bacterial agent BH significantly inhibited the emission of NH₃ in mesophilic phase, and the deodorization effect was better than that of the commercial bacterial agent. After entering thermophilic phase, the deodorization effect of commercial bacterial agent was better than that of bacterial agent BH.【Conclusion】 Two strains of drug-resistant deodorizing bacteria were screened and the removal rates of NH₃ were 48.3% (BH11) and 51.0% (BH12), respectively, and the removal rate of NH₃ by the compound bacterial agent BH was 65.8%. The application of bacterial agent BH to chicken manure composting could effectively reduce the emission of NH₃ during composting, and all indicators met the standards of composting maturity.

Key words: deodorizing bacteria, drug resistance, screening, compound bacterial agent, chicken manure, composting

魏启航, 冯瑶, 王晓醒, 朱宏岗, 方昭, 李兆君. 除臭菌的筛选及其在堆肥中的应用效果[J]. 中国农业科学, 2024, 57(13): 2623-2634.

WEI QiHang, FENG Yao, WANG XiaoXing, ZHU HongGang, FANG Zhao, LI ZhaoJun. Screening of Deodorizing Bacteria and Its Application in Composting[J]. Scientia Agricultura Sinica, 2024, 57(13): 2623-2634.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2024.13.010

https://www.chinaagrisci.com/CN/Y2024/V57/I13/2623

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指标 Index	pH	EC (mS·cm^-1)	含水率 Moisture content (%)	有机碳 Organic carbon (%)	全氮 Total nitrogen (%)
鸡粪 Chicken manure	6.55	3.86	54.07	28.97	3.83
玉米秸秆 Corn straw	/	/	9.69	49.81	0.64

菌株Strain	BH2	BH5	BH9	BH11	BH12	BH15
BH2	-
BH5	-	-
BH9	-	-	-
BH11	-	-	-	-
BH12	-	-	-	-	-
BH15	-	-	-	-	-	-

处理Treatment	庆大霉素含量Gentamicin content		降解率 Degradation rate (%)
处理Treatment	1 d (mg·kg^-1)	40 d (mg·kg^-1)	降解率 Degradation rate (%)
CK	38.76±1.50	19.79±0.61	48.88±2.65a
DT1	38.57±1.62	19.51±0.49	49.40±0.86a
DT2	38.78±0.91	17.13±0.72	55.82±1.47b