整合宏基因组学分析生猪养殖粪污发酵过程中耐药基因的动态变化  

Analysis of dynamic changes of antibiotic resistant genes during pig manure fermentation by integrating metagenomics

作　　者：赵素君[1,2] 曹冶 李江凌[1,2] 张金灵 刘锐[1,2] 王秋实 于吉锋[1,2] ZHAO Su-jun;CAO Ye;LI Jiang-ling;ZHANG Jin-ling;LIU Rui;WANG Qiu-shi;YU Ji-feng(Sichuan Animal Science Academy,Chengdu 610066,China;Animal Breeding and Genetics key Laboratory of Sichuan Province,Chengdu 610066,China)

机构地区：[1]四川省畜牧科学研究院,成都610066 [2]动物遗传育种四川省重点实验室,成都610066

出　　处：《西南农业学报》2023年第3期655-664,共10页Southwest China Journal of Agricultural Sciences

基　　金：中央引导地方科技发展专项自由探索类基础研究(2020ZYD031);国家现代农业产业技术体系四川生猪创新团队粪污有害物迁移途径与环境影响研究岗位(sccxtd-2022-08);四川省科技计划(重点研发项目)(2021YFYZ0030);四川省财政运行专项(SASA2022CZYX007)。

摘　　要：【目的】探究生猪养殖粪污在堆肥过程中耐药基因的消减规律。【方法】以猪粪和菌渣为发酵原料,进行有氧堆肥处理,在堆肥第1、4、8、12、16、22、29天,进行上、中、下分层多点采样,提取总DNA构建文库,经定量和检测合格后,进行宏基因组测序分析。通过宏基因组学方法对堆肥过程中耐药基因的动态变化进行分析。【结果】堆肥期间共发现613个耐药基因,分别对32类抗生素产生耐药。其中,丰度较高的抗生素分别为青霉烷类抗生素(Penam)、头孢菌素类抗生素(Cephalosporin)、氨基糖苷类抗生素(Aminoglycoside antibiotic)、磷霉素类抗生素(Fosfomycin)、糖肽类抗生素(Glycopeptide antibiotic)和四环素类抗生素(Tetracycline antibiotic),这与现实中常用的抗生素种类相吻合。分析发现耐药基因的消减出现2个峰值,第一个峰值出现在试验起始时,在第4天迅速下降,然后在第12天(对照组)和第8天(实验组)达到第二个峰值,随着细菌的自然凋亡,耐药基因的丰度又下降到低值,慢慢消减下去。【结论】在堆肥过程中,生猪养殖粪污中耐药基因丰度呈现波动式消减。【Objective】The paper aimed to study the reduction of antibiotic resistance genes(ARGs)in pig manure during composting.【Method】Pig manure and mushroom residue were used as fermentation materials for aerobic composting.On the 1st,4th,8th,12th,16th,22ndand 29thday of composting,the multi⁃point sampling was carried out in upper,middle and lower layers to extract total DNA and con⁃struct the library.Macrogenomic sequencing analysis was carried out after quantification and qualified detection.The dynamic changes of ARGs during composting were analyzed by metagenomic sequencing.【Result】A total of 613 ARGs were found during composting,which were resistant to 32 classes of antibiotics.Among them,Penam,Cephalosporin,Aminoglycoside,Fosfomycin,Glycopeptide and Tetracycline was more abundant.This was consistent with the types of antibiotics commonly used in reality.Analysis found two peaks in depletion of ARGs.The first peak appeared at the beginning of the trial,declined rapidly on the 4thday,then reached a second peak on the 12th(control group)and 8thday(experimental group).With the natural apoptosis of bacteria,the abundance of ARGs dropped to a low value and gradu⁃ally decreased.【Conclusion】During composting,the ARGs in pig manure decreased in a fluctuating manner.

关 键 词：宏基因组学 粪污 发酵 耐药基因 

分 类 号：S19[农业科学—农业基础科学]