Diverse and abundant antibiotic resistance genes in Chinese swine farms

Antibiotic resistance genes (ARGs) are emerging contaminants posing a potential worldwide human health risk. Intensive animal husbandry is believed to be a major contributor to the increased environmental burden of ARGs. Despite the volume of antibiotics used in China, little information is available regarding the corresponding ARGs associated with animal farms. We assessed type and concentrations of ARGs at three stages of manure processing to land disposal at three large-scale (10,000 animals per year) commercial swine farms in China. In-feed or therapeutic antibiotics used on these farms include all major classes of antibiotics except vancomycins. High-capacity quantitative PCR arrays detected 149 unique resistance genes among all of the farm samples, the top 63 ARGs being enriched 192-fold (median) up to 28,000-fold (maximum) compared with their respective antibiotic-free manure or soil controls. Antibiotics and heavy metals used as feed supplements were elevated in the manures, suggesting the potential for coselection of resistance traits. The potential for horizontal transfer of ARGs because of transposon-specific ARGs is implicated by the enrichment of transposases—the top six alleles being enriched 189-fold (median) up to 90,000-fold in manure—as well as the high correlation (r2 = 0.96) between ARG and transposase abundance. In addition, abundance of ARGs correlated directly with antibiotic and metal concentrations, indicating their importance in selection of resistance genes. Diverse, abundant, and potentially mobile ARGs in farm samples suggest that unmonitored use of antibiotics and metals is causing the emergence and release of ARGs to the environment.

[1]  S. Mindlin,et al.  Tn5045, a novel integron-containing antibiotic and chromate resistance transposon isolated from a permafrost bacterium. , 2011, Research in microbiology.

[2]  Mara Hvistendahl,et al.  Public health. China takes aim at rampant antibiotic resistance. , 2012, Science.

[3]  V. Miriagou,et al.  IS26-Associated In4-Type Integrons Forming Multiresistance Loci in Enterobacterial Plasmids , 2005, Antimicrobial Agents and Chemotherapy.

[4]  S. Levy,et al.  Changes in intestinal flora of farm personnel after introduction of a tetracycline-supplemented feed on a farm. , 1976, The New England journal of medicine.

[5]  F. Michel,et al.  Development and Application of Real-Time PCR Assays for Quantification of erm Genes Conferring Resistance to Macrolides-Lincosamides-Streptogramin B in Livestock Manure and Manure Management Systems , 2007, Applied and Environmental Microbiology.

[6]  S. Koike,et al.  Fate and transport of antibiotic residues and antibiotic resistance genes following land application of manure waste. , 2009, Journal of environmental quality.

[7]  M. Gillings,et al.  Are humans increasing bacterial evolvability? , 2012, Trends in ecology & evolution.

[8]  N. Khardori In-feed antibiotic effects on the swine intestinal microbiome , 2012 .

[9]  S. Levy,et al.  Food Animals and Antimicrobials: Impacts on Human Health , 2011, Clinical Microbiology Reviews.

[10]  Jian-jun Wu,et al.  Potential risks of copper, zinc, and cadmium pollution due to pig manure application in a soil-rice system under intensive farming: a case study of Nanhu, China. , 2011, Journal of environmental quality.

[11]  Satish C. Gupta,et al.  Antibiotic degradation during manure composting. , 2008, Journal of environmental quality.

[12]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[13]  B. Murray,et al.  Antibiotic-resistant bugs in the 21st century--a clinical super-challenge. , 2009, The New England journal of medicine.

[14]  Yong-guan Zhu,et al.  Abundance and diversity of tetracycline resistance genes in soils adjacent to representative swine feedlots in China. , 2010, Environmental science & technology.

[15]  Miriam Barlow,et al.  What antimicrobial resistance has taught us about horizontal gene transfer. , 2009, Methods in molecular biology.

[16]  Natasha Gilbert Rules tighten on use of antibiotics on farms , 2012, Nature.

[17]  Camilla Rodrigues,et al.  Totally drug-resistant tuberculosis in India. , 2012, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[18]  A. Rajić,et al.  Associations between reported on-farm antimicrobial use practices and observed antimicrobial resistance in generic fecal Escherichia coli isolated from Alberta finishing swine farms. , 2009, Preventive veterinary medicine.

[19]  Frank Møller Aarestrup,et al.  Effect of Abolishment of the Use of Antimicrobial Agents for Growth Promotion on Occurrence of Antimicrobial Resistance in Fecal Enterococci from Food Animals in Denmark , 2001, Antimicrobial Agents and Chemotherapy.

[20]  Maliha Aziz,et al.  Staphylococcus aureus CC398: Host Adaptation and Emergence of Methicillin Resistance in Livestock , 2012, mBio.

[21]  H. Heuer,et al.  Plasmids foster diversification and adaptation of bacterial populations in soil. , 2012, FEMS microbiology reviews.

[22]  N. Bolan,et al.  Distribution and bioavailability of copper in farm effluent. , 2003, The Science of the total environment.

[23]  G. Dantas,et al.  The Shared Antibiotic Resistome of Soil Bacteria and Human Pathogens , 2012, Science.

[24]  K. Jones,et al.  Assessment of organic contanhnant fate in waste water treatment plants I: Selected compounds and physicochemical properties , 1999 .

[25]  Otto X. Cordero,et al.  Ecology drives a global network of gene exchange connecting the human microbiome , 2011, Nature.

[26]  J. Weissenbach,et al.  Comparative Genomics of Multidrug Resistance in Acinetobacter baumannii , 2006, PLoS genetics.

[27]  K. Jones,et al.  Assessment of organic contaminant fate in waste water treatment plants. I: Selected compounds and physicochemical properties. , 1999, Chemosphere.

[28]  Jian-Qiang Su,et al.  Fate of tetracyclines in swine manure of three selected swine farms in China. , 2012, Journal of environmental sciences.

[29]  Mazdak Arabi,et al.  Correlation between upstream human activities and riverine antibiotic resistance genes. , 2012, Environmental science & technology.

[30]  Jessica G. Davis,et al.  Response of antibiotics and resistance genes to high-intensity and low-intensity manure management. , 2007, Journal of environmental quality.

[31]  H. Heuer,et al.  IncP-1ε Plasmids are Important Vectors of Antibiotic Resistance Genes in Agricultural Systems: Diversification Driven by Class 1 Integron Gene Cassettes , 2011, Front. Microbio..

[32]  Z. Qiang,et al.  Residual veterinary antibiotics in swine manure from concentrated animal feeding operations in Shandong Province, China. , 2011, Chemosphere.

[33]  Charles W. Knapp,et al.  Evidence of increasing antibiotic resistance gene abundances in archived soils since 1940. , 2010, Environmental science & technology.

[34]  Ramunas Stepanauskas,et al.  Co-selection of antibiotic and metal resistance. , 2006, Trends in microbiology.

[35]  O. Nybroe,et al.  Cu exposure under field conditions coselects for antibiotic resistance as determined by a novel cultivation-independent bacterial community tolerance assay. , 2010, Environmental science & technology.

[36]  S. Ladely,et al.  Effects of Tylosin Use on Erythromycin Resistance in Enterococci Isolated from Swine , 2004, Applied and Environmental Microbiology.

[37]  H. Heuer,et al.  Piggery manure used for soil fertilization is a reservoir for transferable antibiotic resistance plasmids. , 2008, FEMS microbiology ecology.

[38]  C. Knapp,et al.  Abundance of six tetracycline resistance genes in wastewater lagoons at cattle feedlots with different antibiotic use strategies. , 2007, Environmental microbiology.

[39]  M. Chandler,et al.  Insertion Sequences , 1998, Microbiology and Molecular Biology Reviews.

[40]  K. Holmström,et al.  A field survey of chemicals and biological products used in shrimp farming. , 2003, Marine pollution bulletin.

[41]  H. Heuer,et al.  Diverse aadA gene cassettes on class 1 integrons introduced into soil via spread manure. , 2009, Research in microbiology.

[42]  M. Woodward,et al.  A high prevalence of antimicrobial resistant Escherichia coli isolated from pigs and a low prevalence of antimicrobial resistant E. coli from cattle and sheep in Great Britain at slaughter. , 2008, FEMS microbiology letters.

[43]  Heike Schmitt,et al.  Antibiotic resistance gene spread due to manure application on agricultural fields. , 2011, Current opinion in microbiology.

[44]  S. Levy Emergence of antibiotic-resistant bacteria in the intestinal flora of farm inhabitants. , 1978, The Journal of infectious diseases.

[45]  Sudeshna Ghosh,et al.  The effects of subtherapeutic antibiotic use in farm animals on the proliferation and persistence of antibiotic resistance among soil bacteria , 2007, The ISME Journal.

[46]  D. Hughes,et al.  Sampling the Antibiotic Resistome , 2006, Science.

[47]  J. Tiedje,et al.  DNA recovery from soils of diverse composition , 1996, Applied and environmental microbiology.

[48]  Tong Zhang,et al.  Plasmid Metagenome Reveals High Levels of Antibiotic Resistance Genes and Mobile Genetic Elements in Activated Sludge , 2011, PloS one.

[49]  Jessica G. Davis,et al.  tet and sul antibiotic resistance genes in livestock lagoons of various operation type, configuration, and antibiotic occurrence. , 2010, Environmental science & technology.

[50]  P. McDermott,et al.  Identification of antimicrobial resistance and class 1 integrons in Shiga toxin-producing Escherichia coli recovered from humans and food animals. , 2005, The Journal of antimicrobial chemotherapy.

[51]  H. Heuer,et al.  Spreading antibiotic resistance through spread manure: characteristics of a novel plasmid type with low %G+C content. , 2009, Environmental microbiology.

[52]  Zhang Fu-suo,et al.  The estimation of the production amount of animal manure and its environmental effect in China , 2006 .

[53]  D. Church Major factors affecting the emergence and re-emergence of infectious diseases , 2004, Clinics in Laboratory Medicine.