王健;贲伟伟;强志民;潘寻;

• 1:中国科学院生态环境研究中心中国科学院饮用水科学与技术重点实验室
• 2:环境保护部环境保护对外合作中心

摘要(Abstract)：

我国是世界养猪第一大国,生猪饲养量和猪肉产量均位居世界第一。养猪业每年所产生的粪便、废水中含有大量畜用抗生素及其代谢产物,使养猪业废弃物成为环境中重要的抗生素污染源之一,随之产生的抗性基因污染及传播问题也不容忽视。本文结合近年来国内外的研究数据,对我国养猪业废弃物中四环素类、磺胺类抗生素及其相关抗性基因的检测方法、污染状况及影响抗性基因传播的因素进行了分析,并基于控制我国养猪行业抗生素及抗性基因污染的目的,提出了今后的研究重点。

关键词(KeyWords)： 抗生素;抗性基因;四环素;磺胺;猪粪;养猪废水

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(21107127)

作者(Author): 王健;贲伟伟;强志民;潘寻;

Email:

参考文献(References)：

• [1]李振,王云建.畜禽养殖中抗生素使用的现状、问题及对策[J].今日畜牧兽医,2009(8):1-3
• [2]Zhang Q Q,Ying G G,Pan C G,et al.A comprehensive evaluation of antibiotics emission and fate in the river basins of China:Source analysis,multimedia modelling,and linkage to bacterial resistance[J].Environmental Science&Technology,2015,49(11):6772-6782
• [3]Sarmah A K,Meyer M T,Boxall A B A.A global perspective on the use,sales,exposure pathways,occurrence,fate and effects of veterinary antibiotics(VAs)in the environment[J].Chemosphere,2006,65(5):725-759
• [4]周启星,罗义,王美娥.抗生素的环境残留、生态毒性及抗性基因污染[J].生态毒理学报,2007,2(3):243-251Zhou Q X,Luo Y,Wang M E.Environmental residues and ecotoxicity of antibiotics and their resistance gene pollution:A review[J].Asian Journal of Ecotoxicology,2007,2(3):243-251(in Chinese)
• [5]Díaz-Cruz M S,García-Galán M J,BarcelóD.Highly sensitive simultaneous determination of sulfonamide antibiotics and one metabolite in environmental waters by liquid chromatography-quadrupole linear ion trap-mass spectrometry[J].Journal of Chromatography A,2008,1193(1):50-59
• [6]Pan X,Qiang Z M,Ben W W,et al.Simultaneous determination of three classes of antibiotics in the suspended solids of swine wastewater by ultrasonic extraction,solidphase extraction and liquid chromatography-mass spectrometry[J].Journal of Environmental Sciences,2011,23(10):1729-1737
• [7]Kim S,Eichhorn P,Jensen J N,et al.Removal of antibiotics in wastewater:Effect of hydraulic and solid retention times on the fate of tetracycline in the activated sludge process[J].Environmental Science&Technology,2005,39(15):5816-5823
• [8]Tagiri-Endo M,Suzuki S,Nakamura T,et al.Rapid determination of five antibiotic residues in swine wastewater by online solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry[J].Analytical and Bioanalytical Chemistry,2009,393(4):1367-1375
• [9]Jacobsen A M,Halling-Srensen B,Ingerslev F,et al.Simultaneous extraction of tetracycline,macrolide and sulfonamide antibiotics from agricultural soils using pressurised liquid extraction,followed by solid-phase extraction and liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2004,1038(1):157-170
• [10]Ben W W,Qiang Z M,Adams C,et al.Simultaneous determination of sulfonamides,tetracyclines and tiamulin in swine wastewater by solid-phase extraction and liquid chromatography-mass spectrometry[J].Journal of Chromatography A,2008,1202(2):173-180
• [11]Pan X,Qiang Z M,Ben W W,et al.Residual veterinary antibiotics in swine manure from concentrated animal feeding operations in Shandong Province,China[J].Chemosphere,2011,84(5):695-700
• [12]Lu L M,Dai L,Wang Y,et al.Characterization of antimicrobial resistance and integrons among Escherichia coli isolated from animal farms in Eastern China[J].Acta Tropica,2010,113(1):20-25
• [13]Zhang X Y,Ding L J,Yue J.Occurrence and characteristics of class 1 and class 2 integrons in resistant Escherich-ia coli isolates from animals and farm workers in Northeastern China[J].Microbial Drug Resistance,2009,15(4):323-328
• [14]Srinivasan V,Nam H M,Sawant A,et al.Distribution of tetracycline and streptomycin resistance genes and class 1integrons in Enterobacteriaceae isolated from dairy and nondairy farm soils[J].Microbial Ecology,2008,55(2):184-193
• [15]Amann R I,Ludwig W,Schleifer K H.Phylogenetic identification and in situ detection of individual microbial cells without cultivation[J].Microbiological Reviews,1995,59(1):143-169
• [16]Wu N,Qiao M,Zhang B,et al.Abundance and diversity of tetracycline resistance genes in soils adjacent to representative swine feedlots in China[J].Environmental Science&Technology,2010,44(18):6933-6939
• [17]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,2013,110(9):3435-3440
• [18]Mckinney C W,Loftin K A,Meyer M T,et al.tet and sul antibiotic resistance genes in livestock lagoons of various operation type,configuration,and antibiotic occurrence[J].Environmental Science&Technology,2010,44(16):6102-6109
• [19]Heuer H,Smalla K.Manure and sulfadiazine synergistically increased bacterial antibiotic resistance in soil over at least two months[J].Environmental Microbiology,2007,9(3):657-666
• [20]Musovic S,Oregaard G,Kroer N,et al.Cultivation-independent examination of horizontal transfer and host range of an Inc P-1 plasmid among gram-positive and gram-negative bacteria indigenous to the barley rhizosphere[J].Applied and Environmental Microbiology,2006,72(10):6687-6692
• [21]Selvam A,Xu D L,Zhao Z Y,et al.Fate of tetracycline,sulfonamide and fluoroquinolone resistance genes and the changes in bacterial diversity during composting of swine manure[J].Bioresource Technology,2012,126:383-390
• [22]Barkovskii A L,Manoylov K M,Bridges C.Positive and negative selection towards tetracycline resistance genes in manure treatment lagoons[J].Journal of Applied Microbiology,2012,112(5):907-919
• [23]Peak N,Knapp C W,Yang R K,et al.Abundance of six tetracycline resistance genes in wastewater lagoons at cattle feedlots with different antibiotic use strategies[J].Environmental Microbiology,2007,9(1):143-151
• [24]Aminov R I,Garrigues-Jeanjean N,Mackie R I.Molecular ecology of tetracycline resistance:Development and validation of primers for detection of tetracycline resistance genes encoding ribosomal protection proteins[J].Applied and Environmental Microbiology,2001,67(1):22-32
• [25]Luo Y,Mao D Q,Rysz M,et al.Trends in antibiotic resistance genes occurrence in the Haihe River,China[J].Environmental Science&Technology,2010,44(19):7220-7225
• [26]Barkovskii A,Bridges C.Persistence and profiles of tetracycline resistance genes in swine farms and impact of operational practices on their occurrence in farms'vicinities[J].Water,Air,&Soil Pollution,2012,223(1):49-62
• [27]Zhang X X,Zhang T,Fang H.Antibiotic resistance genes in water environment[J].Applied Microbiology and Biotechnology,2009,82(3):397-414
• [28]Ng L K,Martin I,Alfa M,et al.Multiplex PCR for the detection of tetracycline resistant genes[J].Molecular and Cellular Probes,2001,15(4):209-215
• [29]Garofalo C,Vignaroli C,Zandri G,et al.Direct detection of antibiotic resistance genes in specimens of chicken and pork meat[J].International Journal of Food Microbiology,2007,113(1):75-83
• [30]Khan S A,Nawaz M S,Khan A A,et al.Molecular characterization of multidrug-resistant Enterococcus spp.from poultry and dairy farms:Detection of virulence and vancomycin resistance gene markers by PCR[J].Molecular and Cellular Probes,2005,19(1):27-34
• [31]Moura A,Henriques I,Ribeiro R,et al.Prevalence and characterization of integrons from bacteria isolated from a slaughterhouse wastewater treatment plant[J].Journal of Antimicrobial Chemotherapy,2007,60(6):1243-1250
• [32]Perreten V,Vorlet-Fawer L,Slickers P,et al.Microarraybased detection of 90 antibiotic resistance genes of grampositive bacteria[J].Journal of Clinical Microbiology,2005,43(5):2291-2302
• [33]Frye J G,Jesse T,Long F,et al.DNA microarray detection of antimicrobial resistance genes in diverse bacteria[J].International Journal of Antimicrobial Agents,2006,27(2):138-151
• [34]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
• [35]Nyrén P,Karamohamed S,Ronaghi M.Detection of single-base changes using a biolumino-metric primer extension assay[J].Analytical Biochemistry,1997,244(2):367-373
• [36]Ronaghi M,Karamohamed S,Pettersson B,et al.Realtime DNA sequencing using detection of pyrophosphate release[J].Analytical Biochemistry,1996,242(1):84-89
• [37]Shendure J,Ji H.Next-generation DNA sequencing[J].Nature Biotechnology,2008,26(10):1135-1145
• [38]Allen H K.Antibiotic resistance gene discovery in foodproducing animals[J].Current Opinion in Microbiology,2014,19:25-29
• [39]Christgen B,Yang Y,Ahammad S Z,et al.Metagenomics shows that low-energy anaerobic-aerobic treatment reactors reduce antibiotic resistance gene levels from domestic wastewater[J].Environmental Science&Technology,2015,49(4):2577-2584
• [40]Yang Y,Li B,Ju F,et al.Exploring variation of antibiotic resistance genes in activated sludge over a four-year period through a metagenomic approach[J].Environmental Science&Technology,2013,47(18):10197-10205
• [41]Sydenham T V,Sóki J,Hasman H,et al.Identification of antimicrobial resistance genes in multidrug-resistant clinical Bacteroides fragilis isolates by whole genome shotgun sequencing[J].Anaerobe,2015,31:59-64
• [42]Schmieder R,Edwards R.Insights into antibiotic resistance through metagenomic approaches[J].Future Microbiology,2011,7(1):73-89
• [43]张慧敏,章明奎,顾国平.浙北地区畜禽粪便和农田土壤中四环素类抗生素残留[J].生态与农村环境学报,2008,24(3):69-73Zhang H M,Zhang M K,Gu G P.Residues of tetracyclines in livestock and poultry manures and agricultural soils from North Zhejiang Province[J].Journal of Ecology and Rural Environment,2008,24(3):69-73(in Chinese)
• [44]张树清,张夫道,刘秀梅,等.规模化养殖畜禽粪主要有害成分测定分析研究[J].植物营养与肥料学报,2005,11(6):822-829Zhang S Q,Zhang F D,Liu X M,et al.Determination and analysis on main harmful composition in excrement of scale livestock and poultry feedlots[J].Plant Nutrition and Fertilizer Science,2005,11(6):822-829(in Chinese)
• [45]刘新程,董元华,王辉.江苏省集约化养殖畜禽排泄物中四环素类抗生素残留调查[J].农业环境科学学报,2008,27(3):1177-1182Liu X C,Dong Y H,Wang H.Residues of tetracyclines in animal manure from intensive farm in Jiangsu Province[J].Journal of Agro-Environment Science,2008,27(3):1177-1182(in Chinese)
• [46]魏瑞成,王冉,李维,等.猪粪中金霉素残留的测定方法[J].浙江农业学报,2008,20(4):291-295Wei R C,Wang R,Li W,et al.Determination method of chlortetracycline residues in pig faeces[J].Acta Agriculturae Zhejiangensis,2008,20(4):291-295(in Chinese)
• [47]Hu X G,Luo Y,Zhou Q X,et al.Determination of thirteen antibiotics residues in manure by solid phase extraction and high performance liquid chromatography[J].Chinese Journal of Analytical Chemistry,2008,36(9):1162-1166
• [48]Martínez-Carballo E,González-Barreiro C,Scharf S,et al.Environmental monitoring study of selected veterinary antibiotics in animal manure and soils in Austria[J].Environmental Pollution,2007,148(2):570-579
• [49]Jacobsen A M,Halling-Srensen B.Multi-component analysis of tetracyclines,sulfonamides and tylosin in swine manure by liquid chromatography-tandem mass spectrometry[J].Analytical and Bioanalytical Chemistry,2006,384(5):1164-1174
• [50]Zhao L,Dong Y H,Wang H.Residues of veterinary antibiotics in manures from feedlot livestock in eight provinces of China[J].Science of the Total Environment,2010,408(5):1069-1075
• [51]胡献刚,罗义,周启星,等.固相萃取-高效液相色谱法测定畜牧粪便中13种抗生素药物残留[J].分析化学,2008,36(9):1162-1166Hu X G,Luo Y,Zhou Q X,et al.Determination of thirteen antibiotics residues in manure by solid phase extraction and high performance liquid chromatography[J].Chinese Journal of Analytical Chemistry,2008,36(9):1162-1166(in Chinese)
• [52]Haller M Y,Müller S R,Mcardell C S,et al.Quantification of veterinary antibiotics(sulfonamides and trimethoprim)in animal manure by liquid chromatographymass spectrometry[J].Journal of Chromatography A,2002,952(1-2):111-120
• [53]Wei R C,Ge F,Huang S Y,et al.Occurrence of veterinary antibiotics in animal wastewater and surface water around farms in Jiangsu Province,China[J].Chemosphere,2011,82(10):1408-1414
• [54]Tong L,Li P,Wang Y X,et al.Analysis of veterinary antibiotic residues in swine wastewater and environmental water samples using optimized SPE-LC/MS/MS[J].Chemosphere,2009,74(8):1090-1097
• [55]Ben W W,Pan X,Qiang Z M.Occurrence and partition of antibiotics in the liquid and solid phases of swine wastewater from concentrated animal feeding operations in Shandong Province,China[J].Environmental Science:Processes&Impacts,2013,15(4):870-875
• [56]Campagnolo E R,Johnson K R,Karpati A,et al.Antimi-crobial residues in animal waste and water resources proximal to large-scale swine and poultry feeding operations[J].Science of the Total Environment,2002,299(1-3):89-95
• [57]Malintan N T,Mohd M A.Determination of sulfonamides in selected Malaysian swine wastewater by high-performance liquid chromatography[J].Journal of Chromatography A,2006,1127(1-2):154-160
• [58]Hu X G,Zhou Q X,Luo Y.Occurrence and source analysis of typical veterinary antibiotics in manure,soil,vegetables and groundwater from organic vegetable bases,Northern China[J].Environmental Pollution,2010,158(9):2992-2998
• [59]Li Y W,Wu X L,Mo C H,et al.Investigation of sulfonamide,tetracycline,and quinolone antibiotics in vegetable farmland soil in the Pearl River Delta Area,Southern China[J].Journal of Agricultural and Food Chemistry,2011,59(13):7268-7276
• [60]Huang X,Liu C X,Li K,et al.Occurrence and distribution of veterinary antibiotics and tetracycline resistance genes in farmland soils around swine feedlots in Fujian Province,China[J].Environmental Science and Pollution Research,2013,20(12):9066-9074
• [61]Pawelzick H,Hoper H,Nau H,et al.A survey of the occurrence of various tetracyclines and sulfamethazine in sandy soils in Northwestern Germany fertilized with liquid manure[C].SETAC Euro 14th Annual Meeting,Prague:Czech Republic,2004:18-22
• [62]Hamscher G,Sczesny S,Hper H,et al.Determination of persistent tetracycline residues in soil fertilized with liquid manure by high-performance liquid chromatography with electrospray ionization tandem mass spectrometry[J].Analytical Chemistry,2002,74(7):1509-1518
• [63]Thaker M,Spanogiannopoulos P,Wright G.The tetracycline resistome[J].Cellular and Molecular Life Sciences,2010,67(3):419-431
• [64]Skld O.Sulfonamide resistance:Mechanisms and trends[J].Drug Resistance Updates,2000,3(3):155-160
• [65]Roberts M C.Update on acquired tetracycline resistance genes[J].FEMS Microbiology Letters,2005,245(2):195-203
• [66]Mu Q H,Li J,Sun Y X,et al.Occurrence of sulfonamide-,tetracycline-,plasmid-mediated quinolone-and macrolide-resistance genes in livestock feedlots in Northern China[J].Environmental Science and Pollution Research,2014,22(9):1-9
• [67]Peng S,Wang Y M,Zhou B B,et al.Long-term application of fresh and composted manure increase tetracycline resistance in the arable soil of Eastern China[J].Science of the Total Environment,2015,506-507:279-286
• [68]Miller R.Environmental bacteriophage-host interactions:Factors contribution to natural transduction[J].Antonie Van Leeuwenhoek,2001,79(2):141-147
• [69]Thomas C M,Nielsen K M.Mechanisms of,and barriers to,horizontal gene transfer between bacteria[J].Nature Reviews Microbiology,2005,3(9):711-721
• [70]Frost L S,Leplae R,Summers A O,et al.Mobile genetic elements:The agents of open source evolution[J].Nature Reviews Microbiology,2005,3(9):722-732
• [71]Norman A,Hansen L H,Srensen S J.Conjugative plasmids:Vessels of the communal gene pool[J].Philosophical Transactions of the Royal Society B,2009,364(1527):2275-2289
• [72]Bennett P M.Plasmid encoded antibiotic resistance:Acquisition and transfer of antibiotic resistance genes in bacteria[J].British Journal of Pharmacology,2008,153(S1):S347-S357
• [73]Binh C T T,Heuer H,Kaupenjohann M,et al.Piggery manure used for soil fertilization is a reservoir for transferable antibiotic resistance plasmids[J].FEMS Microbiology Ecology,2008,66(1):25-37
• [74]Smalla K,Heuer H,Gotz A,et al.Exogenous isolation of antibiotic resistance plasmids from piggery manure slurries reveals a high prevalence and diversity of Inc Q-like plasmids[J].Applied and Environmental Microbiology,2000,66(11):4854-4862
• [75]Liu M M,Zhang Y,Yang M,et al.Abundance and distribution of tetracycline resistance genes and mobile elements in an oxytetracycline production wastewater treatment system[J].Environmental Science&Technology,2012,46(14):7551-7557
• [76]Koike S,Krapac I G,Oliver H D,et al.Monitoring and source tracking of tetracycline resistance genes in lagoons and groundwater adjacent to swine production facilities over a 3-year period[J].Applied and Environmental Microbiology,2007,73(15):4813-4823
• [77]Heuer H,Schmitt H,Smalla K.Antibiotic resistance gene spread due to manure application on agricultural fields[J].Current Opinion in Microbiology,2011,14(3):236-243
• [78]Musovic S,Klümper U,Dechesne A,et al.Long-term manure exposure increases soil bacterial community potential for plasmid uptake[J].Environmental Microbiology Reports,2014,6(2):125-130
• [79]Séveno N A,Kallifidas D,Smalla K,et al.Occurrence and reservoirs of antibiotic resistance genes in the environment[J].Reviews in Medical Microbiology,2002,13(1):15-27
• [80]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
• [81]Selvam A,Zhao Z,Li Y C,et al.Degradation of tetracycline and sulfadiazine during continuous thermophilic composting of pig manure and sawdust[J].Environmental Technology,2013,34(16):1-9
• [82]Selvam A,Zhao Z Y,Wong J W C.Composting of swine manure spiked with sulfadiazine,chlortetracycline and ciprofloxacin[J].Bioresource Technology,2012,126:412-417
• [83]Wu X F,Wei Y S,Zheng J X,et al.The behavior of tetracyclines and their degradation products during swine manure composting[J].Bioresource Technology,2011,102(10):5924-5931
• [84]Arikan O A,Mulbry W,Rice C.Management of antibiotic residues from agricultural sources:Use of composting to reduce chlortetracycline residues in beef manure from treated animals[J].Journal of Hazardous materials,2009,164(2-3):483-489
• [85]Dolliver H,Gupta S,Noll S.Antibiotic degradation during manure composting[J].Journal of Environmental Quality,2008,37(3):1245-1253
• [86]Arikan O A,Sikora L J,Mulbry W,et al.Composting rapidly reduces levels of extractable oxytetracycline in manure from therapeutically treated beef calves[J].Bioresource Technology,2007,98(1):169-176
• [87]潘寻,强志民,贲伟伟.高温堆肥对猪粪中多类抗生素的去除效果[J].生态与农村环境学报,2013,29(1):64-69Pan X,Qiang Z M,Ben W W.Effects of high-temperature composting on degradation of antibiotics in swine manure[J].Journal of Ecology and Rural Environment,2013,29(1):64-69(in Chinese)
• [88]Wang L L,Oda Y,Grewal S,et al.Persistence of resistance to erythromycin and tetracycline in swine manure during simulated composting and lagoon treatments[J].Microbial Ecology,2012,63(1):32-40
• [89]Yu Z T,Michel F C,Hansen G,et al.Development and application of real-time PCR assays for quantification of genes encoding tetracycline resistance[J].Applied and Environmental Microbiology,2005,71(11):6926-6933
• [90]Engemann C A,Keen P L,Knapp C W,et al.Fate of tetracycline resistance genes in aquatic systems:Migration from the water column to peripheral biofilms[J].Environmental Science&Technology,2008,42(14):5131-5136◆