紫色土丘陵区畜禽养殖场土壤中抗生素抗性基因分布特征
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摘要
鉴于紫色土高垦殖丘陵区土壤中抗生素抗性基因的潜在生态环境风险,本研究采用高通量荧光定量PCR技术,分析了抗生素抗性基因在不同类型畜禽养殖场(养猪场、养鸡场和养牛场)土壤中的丰度和多样性.结果表明,3种养殖场土壤中共检出79种抗生素抗性基因和5种可移动基因元件,其中多重耐药类基因是养殖场土壤中丰度最高的耐药基因.不同养殖场土壤中抗生素抗性基因的丰度和多样性均表现为养鸡场和养猪场样品高于养牛场土壤,且3种养殖场土壤样品中的抗性基因表现出不同的类别分布特征.养殖场土壤中较高的可移动基因元件丰度,及其与抗生素抗性基因丰度显著的相关性(P<0. 05)说明,可移动基因元件可能促进了抗生素抗性基因在养殖场土壤中的迁移和扩散.
Given the potential risk posed by antibiotic resistance genes( ARGs) to the eco-environment in the hilly purple soil region,which has been intensively utilized by human,surface soil samples were collected from feedlots of pig,chicken,and cattle farms and were analyzed for the diversity and abundance of ARGs using high-throughput QPCR. In total,79 ARGs and 5 mobile genetic elements( MGEs) were detected across all samples. Among these genes,multidrug resistance genes were the most abundant type of ARGs.More abundant and diverse ARGs were observed in feedlot soil samples from pig and chicken farms than those from cattle feedlot soils,and these samples showed different distribution patterns of ARGs. High abundance of MGEs and their significant correlation with ARGs( P < 0. 05) implied an important role of MGEs in the dissemination of ARGs in livestock feedlot soils.
Given the potential risk posed by antibiotic resistance genes( ARGs) to the eco-environment in the hilly purple soil region,which has been intensively utilized by human,surface soil samples were collected from feedlots of pig,chicken,and cattle farms and were analyzed for the diversity and abundance of ARGs using high-throughput QPCR. In total,79 ARGs and 5 mobile genetic elements( MGEs) were detected across all samples. Among these genes,multidrug resistance genes were the most abundant type of ARGs.More abundant and diverse ARGs were observed in feedlot soil samples from pig and chicken farms than those from cattle feedlot soils,and these samples showed different distribution patterns of ARGs. High abundance of MGEs and their significant correlation with ARGs( P < 0. 05) implied an important role of MGEs in the dissemination of ARGs in livestock feedlot soils.
引文
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[18]张丹丹,郭亚平,任红云,等.福建省敖江下游抗生素抗性基因分布特征[J].环境科学,2018,39(6):2600-2606.Zhang D D,Guo Y P,Ren H Y,et al. Characteristics of antibiotic resistance genes in downstream areas of the Aojiang river,Fujian province[J]. Environmental Science,2018,39(6):2600-2606.
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[21]魏光伟,魏文康,余永鹏,等.饲料抗生素添加剂的应用技术[J].广东饲料,2007,16(3):15-16.Wei G W,Wei W K,Yu Y P,et al. The technique of antibiosis application in Feed[J]. Guangdong Feed,2007,16(3):15-16.
[22] Chen B W,Lin L,Fang L,et al. Complex pollution of antibiotic resistance genes due to beta-lactam and aminoglycoside use in aquaculture farming[J]. Water Research,2018,134:200-208.
[23] Zhang X X,Zhang T,Fang H H P. Antibiotic resistance genes in water environment[J]. Applied Microbiology and Biotechnology,2009,82(3):397-414.
[24] Klappenbach J A,Saxman P R,Cole J R,et al. rrndb:the ribosomal RNA operon copy number database[J]. Nucleic Acids Research,2001,29(1):181-184.
[25] Fang H,Wang H F,Cai L,et al. Prevalence of antibiotic resistance genes and bacterial pathogens in long-term manured greenhouse soils as revealed by metagenomic survey[J].Environmental Science&Technology,2015,49(2):1095-1104.
[26]段曼莉.生物炭对土壤中抗生素及其抗性基因变化的影响研究[D].杨凌:西北农林科技大学,2017. 23-24.Duan M L. Effects of biochar application on variation of antibiotic and antibiotic resistance genes in soil[D]. Yangling:Northwest A&F University,2017. 23-24.
[27] Zhang Q Q, Ying G G, Pan C G, et al. Comprehensive evaluation of antibiotics emission and fate in the river basins of China:source analysis,multimedia modeling,and linkage to bacterial resistance[J]. Environmental Science&Technology,2015,49(11):6772-6782.
[28] Hou J,Wan W N,Mao D Q,et al. Occurrence and distribution of sulfonamides, tetracyclines, quinolones, macrolides, and nitrofurans in livestock manure and amended soils of Northern China[J]. Environmental Science and Pollution Research,2015,22(6):4545-4554.
[29] Dantas G,Sommer M O A. How to fight back against antibiotic resistance[J]. American Scientist,2014,102(1):42-51.
[2]朱永官,欧阳纬莹,吴楠,等.抗生素耐药性的来源与控制对策[J].中国科学院院刊,2015,30(4):509-516.Zhu Y G,Ouyang W Y,Wu N,et al. Antibiotic resistance:sources and mitigation[J]. Bulletin of Chinese Academy of Sciences,2015,30(4):509-516.
[3]苏建强,黄福义,朱永官.环境抗生素抗性基因研究进展[J].生物多样性,2013,21(4):481-487.Su J Q,Huang F Y,Zhu Y G. Antibiotic resistance genes in the environment[J]. Biodiversity Science,2013,21(4):481-487.
[4] D'Costa V M,King C E,Kalan L,et al. Antibiotic resistance is ancient[J]. Nature,2011,477(7365):457-461.
[5] Knapp C W,Dolfing J,Ehlert P A I,et al. Evidence of increasing antibiotic resistance gene abundances in archived soils since 1940[J]. Environmental Science&Technology,2010,44(2):580-587.
[6] Looft T,Johnson T A,Allen H K,et al. In-feed antibiotic effects on the swine intestinal microbiome[J]. Proceedings of the National Academy of Sciences of the United States of America,2012,109(5):1691-1696.
[7] Chee-Sanford J C,Mackie R I,Koike S,et al. Fate and transport of antibiotic residues and antibiotic resistance genes following land application of manure waste[J]. Journal of Environmental Quality,2009,38(3):1086-1108.
[8] Zhu Y G,Johnson T A,Su J Q,et al. Diverse and abundant antibiotic resistance genes in Chinese swine farms[J].Proceedings of the National Academy of Sciences of the United States of America,2013,110(9):3435-3440.
[9] He L Y,Liu Y S,Su H C,et al. Dissemination of antibiotic resistance genes in representative broiler feedlots environments:identification of indicator ARGs and correlations with environmental variables[J]. Environmental Science&Technology,2014,48(22):13120-13129.
[10] Forsberg K J,Reyes A,Wang B,et al. The shared antibiotic resistome of soil bacteria and human pathogens[J]. Science,2012,337(6098):1107-1111.
[11] Cheng W X,Chen H,Su C,et al. Abundance and persistence of antibiotic resistance genes in livestock farms:a comprehensive investigation in eastern China[J]. Environment International,2013,61:1-7.
[12] Qian X,Gu J,Sun W,et al. Diversity,abundance,and persistence of antibiotic resistance genes in various types of animal manure following industrial composting[J]. Journal of Hazardous Materials,2018,344:716-722.
[13] Peng S,Feng Y Z,Wang Y M,et al. Prevalence of antibiotic resistance genes in soils after continually applied with different manure for 30 years[J]. Journal of Hazardous Materials,2017,340:16-25.
[14] Zhou B R, Wang C, Zhao Q, et al. Prevalence and dissemination of antibiotic resistance genes and coselection of heavy metals in Chinese dairy farms[J]. Journal of Hazardous Materials,2016,320:10-17.
[15]仇焕广,廖绍攀,井月,等.我国畜禽粪便污染的区域差异与发展趋势分析[J].环境科学,2013,34(7):2766-2774.Qiu H G,Liao S P,Jing Y,et al. Regional differences and development tendency of livestock manure pollution in China[J].Environmental Science,2013,34(7):2766-2774.
[16] Van Boeckel T P,Brower C,Gilbert M,et al. Global trends in antimicrobial use in food animals[J]. Proceedings of the National Academy of Sciences of the United States of America,2015,112(18):5649-5654.
[17]鲜青松,唐翔宇,朱波.坡耕地薄层紫色土-岩石系统中氮磷的迁移特征[J].环境科学,2017,38(7):2843-2849.Xian Q S, Tang X Y, Zhu B. Transport of nitrogen and phosphorus from sloping farmland with thin purple soil overlying rocks[J]. Environmental Science,2017,38(7):2843-2849.
[18]张丹丹,郭亚平,任红云,等.福建省敖江下游抗生素抗性基因分布特征[J].环境科学,2018,39(6):2600-2606.Zhang D D,Guo Y P,Ren H Y,et al. Characteristics of antibiotic resistance genes in downstream areas of the Aojiang river,Fujian province[J]. Environmental Science,2018,39(6):2600-2606.
[19] Cheng J H,Tang X Y,Cui J F. Effect of long-term manure slurry application on the occurrence of antibiotic resistance genes in arable purple soil(entisol)[J]. Science of the Total Environment,2019,647:853-861.
[20]黄福义,安新丽,陈青林,等.梅花鹿养殖场抗生素抗性基因分布特征[J].环境科学,2016,37(11):4402-4409.Huang F Y, An X L, Chen Q L, et al. Distribution characteristics of antibiotic resistance genes in sika deer farm[J]. Environmental Science,2016,37(11):4402-4409.
[21]魏光伟,魏文康,余永鹏,等.饲料抗生素添加剂的应用技术[J].广东饲料,2007,16(3):15-16.Wei G W,Wei W K,Yu Y P,et al. The technique of antibiosis application in Feed[J]. Guangdong Feed,2007,16(3):15-16.
[22] Chen B W,Lin L,Fang L,et al. Complex pollution of antibiotic resistance genes due to beta-lactam and aminoglycoside use in aquaculture farming[J]. Water Research,2018,134:200-208.
[23] Zhang X X,Zhang T,Fang H H P. Antibiotic resistance genes in water environment[J]. Applied Microbiology and Biotechnology,2009,82(3):397-414.
[24] Klappenbach J A,Saxman P R,Cole J R,et al. rrndb:the ribosomal RNA operon copy number database[J]. Nucleic Acids Research,2001,29(1):181-184.
[25] Fang H,Wang H F,Cai L,et al. Prevalence of antibiotic resistance genes and bacterial pathogens in long-term manured greenhouse soils as revealed by metagenomic survey[J].Environmental Science&Technology,2015,49(2):1095-1104.
[26]段曼莉.生物炭对土壤中抗生素及其抗性基因变化的影响研究[D].杨凌:西北农林科技大学,2017. 23-24.Duan M L. Effects of biochar application on variation of antibiotic and antibiotic resistance genes in soil[D]. Yangling:Northwest A&F University,2017. 23-24.
[27] Zhang Q Q, Ying G G, Pan C G, et al. Comprehensive evaluation of antibiotics emission and fate in the river basins of China:source analysis,multimedia modeling,and linkage to bacterial resistance[J]. Environmental Science&Technology,2015,49(11):6772-6782.
[28] Hou J,Wan W N,Mao D Q,et al. Occurrence and distribution of sulfonamides, tetracyclines, quinolones, macrolides, and nitrofurans in livestock manure and amended soils of Northern China[J]. Environmental Science and Pollution Research,2015,22(6):4545-4554.
[29] Dantas G,Sommer M O A. How to fight back against antibiotic resistance[J]. American Scientist,2014,102(1):42-51.
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