复合菌剂添加对鸡粪膜堆肥过程中耐药基因消减的影响

曾经理; 谢诗茵; 邓毅恒; 牛世华; 李有建; 邢斯程; 廖新俤

文章摘要

曾经理,谢诗茵,邓毅恒,牛世华,李有建,邢斯程,廖新俤.复合菌剂添加对鸡粪膜堆肥过程中耐药基因消减的影响[J].农业环境科学学报,2025,44(6):1636-1646.

复合菌剂添加对鸡粪膜堆肥过程中耐药基因消减的影响

Effects of composite bacterial inocula on the reduction of antibiotic resistance genes during the membrane-covered composting of chicken manure

投稿时间：2024-05-31

DOI：10.11654/jaes.2024-0455

中文关键词: 鸡粪膜堆肥耐药基因复合菌剂分层样本氮素

英文关键词: chicken manure membrane composting ARGs compound bacterial agent stratified sample nitrogen

基金项目:国家自然科学基金项目（32072783）；现代农业产业技术体系建设专项资金项目（CARS-40）

作者	单位	E-mail
曾经理	华南农业大学动物科学学院, 广州 510642
谢诗茵	华南农业大学动物科学学院, 广州 510642
邓毅恒	华南农业大学动物科学学院, 广州 510642
牛世华	温氏食品集团股份有限公司, 广东云浮 527300
李有建	温氏食品集团股份有限公司, 广东云浮 527300
邢斯程	华南农业大学群体微生物研究中心, 广州 510642 广东省农用动物基因组学与分子育种重点实验室, 广州 510642 畜禽繁育国地联合工程技术研究中心, 广州 510642
廖新俤	华南农业大学动物科学学院, 广州 510642 广东省农用动物基因组学与分子育种重点实验室, 广州 510642 畜禽繁育国地联合工程技术研究中心, 广州 510642 广东省猪禽养殖业重点实验室, 广州 510642	xdliao@scau.edu.cn

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中文摘要:

为探究复合菌剂添加对蛋鸡粪膜堆肥过程中耐药基因消减的影响，以蘑菇渣为辅料混合蛋鸡粪开展膜堆肥试验。试验设计空白肉汤组（CK）、市售菌剂组（EM）、自研菌剂组（PB4和PB5），采集第0天和第30天的两个混合样本，并对第30天样本额外采集上、中、下层分层样本，对样本理化指标和耐药基因进行分析，分别探讨3种菌剂对膜堆肥过程中耐药基因的消减规律和对氮素固存的影响。结果表明：本研究所选取的9个耐药基因和2个可移动遗传元件，在第30天混合样本中，相较于初始耐药基因绝对丰度，EM组相较于CK组显著提高了bla-NDM、tetA的去除率（P<0.05），PB4、PB5组相较于CK组显著提高了bla-NDM、ermC、intI2、tetA的去除率（P<0.05），但PB5组对drfA1、intI1、tetX的去除效果显著低于CK组（P<0.05）。相较于初始丰度，PB5菌剂还促进了tetX丰度的增加，EM菌剂促进了ermC、intI2、tetX丰度的升高，CK组中ermC、intI2、tetA丰度也有升高。对比组内初始耐药基因丰度，添加PB4菌剂对总耐药基因绝对丰度的去除率为77.63%，显著高于CK（67.31%）、EM（70.36%）和PB5（72.23%）。分层样本中，EM菌剂对下层样本耐药基因去除效果最佳，去除率为53.15%，PB4、PB5菌剂对上、中层样本耐药基因去除率均高于90%，但PB5菌剂提高了下层样本耐药基因的丰度。PB4菌剂在氮素固存方面表现最佳，相较堆肥结束时的CK组，添加PB4菌剂显著增加了13.72%的铵态氮含量，显著减少了11.97%的氨气累计排放量（P<0.05），添加EM菌剂和PB5菌剂对氨气减排效果不显著，且添加PB5菌剂使得硝态氮含量降低。研究表明，PB4菌剂对于耐药基因的消减以及氮素固存的综合效益最佳。

英文摘要:

To explore the pattern of antibiotic resistance genes（ARGs）reduction during the composting process of laying hen manur, mushroom residue was used as auxiliary material to mix with laying hens manure for conducting a composting test. The experimental design included a blank broth group（CK）, a commercial bacterial agent group（EM）, and two self-developed bacterial agent groups（PB4 andPB5）. Mixed samples were collected on days 0 and 30, and samples from the top, middle, and bottom layers were collected on day 30. The physical and chemical indexes and ARGs of the samples were analyzed, and the effects of three bacterial agents on the reduction of ARGs and nitrogen fixation during membrane composting were discussed. The results indicate that among the nine selected resistance genes and two mobile genetic elements in this study, in the mixed samples on day 30, compared with the initial absolute abundance of ARGs, the removal rates of bla-NDM and tetA in the EM group were significantly increased compared with the CK group（P<0.05）. Additionally, the removal rates of bla-NDM, ermC, intI2 and tetA in PB4 and PB5 groups were also significantly increased compared with the CK group（P< 0.05）. However, it was found that the removal effect of drfA1, intI1 and tetX in PB5 group was significantly lower than that in the CK group （P<0.05）. Furthermore, when comparing with their initial abundance levels, it was observed that using PB5 bacteria agent promoted an increase in tetX abundance. Similarly, using the EM bacteria agent led to an increase in the abundance of ermC, intI2 and tetX. The abundance of ermC, intI2 and tetA also increased in the CK group. Comparing the initial abundance of resistance genes within each group, the addition of PB4 bacteria agent removed 77.63% of the absolute abundance of total resistance genes, which was significantly higher than that of CK（67.31%）, EM（70.36%）and PB5（72.23%）. In stratified samples, EM bacteria agent exhibited the highest removal efficiency of antibiotic resistance genes（ARGs）in the bottom layer at 53.15%, while PB4 and PB5 bacteria agents achieved removal rates exceeding 90% in the upper and middle layers. However, PB5 bacteria agent increased the abundance of ARGs in the bottom layer. After composting, the addition of PB4 bacterial agent significantly increased the ammonium nitrogen content by 13.72%, while also reducing accumulated ammonia emission by 11.97%（P<0.05）compared to the CK group. However, the addition of EM bacteria agent and PB5 bacterial agent did not have a significant effect on reducing NH₃ emission. Additionally, the content of nitrate nitrogen decreased with the addition of PB5 bacteria agent. In summary, the PB4 bacteria agent has the best comprehensive benefit for the reduction of ARGs and nitrogen fixation.

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