Abstract
Class 1 integronase gene (intI1) and tetracycline resistance genes (tetA and tetC) from various environmental sites in Jiangsu Province (China) were detected using qualitative PCR (polymerase chain reaction) and quantified with SYBR Green-based qRT-PCR (quantitative real-time PCR) in this study. Qualitative PCR assays demonstrated that intI1, tetA and tetC occurred in the water environments of Taihu Lake, the Nanjing section of the Yangtze River, a sewage treatment plant (STP) in Nanjing City, and two drinking water treating bioreactors. qRT-PCR results showed that abundance of intI1 in lake water and sediments was lower than the tet genes, for a given sample site and date (P < 0.05). On a volumetric basis, lake sediments contained higher concentrations of the three genes by four to five orders of magnitude than water samples, and lake water and sediments sampled in April contained fewer copies of all the genes than the samples collected in June and August (P < 0.05). The levels of intI1, tetA and tetC in the Yangtze River water increased significantly after the river flowed through Nanjing City (P < 0.05). 94.1% integron, 97.2% tetA and 98.3% tetC were removed by the activated sludge process in the STP, and more than 80% of each gene was removed in both of the two biofilters in terms of relative concentration based on sample volume. However, on the basis of DNA mass, lower removals were obtained for both the activated sludge and biofiltration processes.
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Akinbowale OL, Peng H, Barton MD (2007) Class 1 integron mediates antibiotic resistance in Aeromonas spp. from rainbow trout farms in Australia. Int J Antimicrob Agents 29:S113
Aminov RI, Mackie RI (2007) Evolution and ecology of antibiotic resistance genes. FEMS Microbiol Lett 271:147–161
Aminov RI, Chee-Sanford JC, Garrigues N, Teferedegne B, Krapac IJ, White BA, Mackie RI (2002) Development, validation, and application of PCR primers for detection of tetracycline efflux genes of gram-negative bacteria. Appl Environ Microbiol 68:1786–1793
Auerbach EA, Seyfried EE, McMahon KD (2007) Tetracycline resistance genes in activated sludge wastewater treatment plants. Water Res 41:1143–1151
Bruce JR, Paul KSL, Michael M (2005) Emerging chemicals of concern: pharmaceuticals and personal care products (PPCPs) in Asia, with particular reference to Southern China. Mar Pollut Bull 50:913–920
Chee-Sanford JC, Aminov RI, Krapac IJ, Garrigues-Jeanjean N, Mackie RI (2001) Occurrence and diversity of tetracycline resistance genes in lagoons and groundwater underlying two swine production facilities. Appl Environ Microbiol 67:1494–1502
Dalsgaard A, Forslund A, Serichantalergs O, Sandvang D (2000) Distribution and content of class 1 integrons in different Vibrio cholerae O-serotype strains isolated in Thailand. Antimicrob Agents Chemother 44:1315–1321
Dang HY, Zhang XX, Song LS, Chang YQ, Yang GP (2007) Molecular determination of oxytetracycline-resistant bacteria and their resistance genes from mariculture environments of China. J Appl Microbiol 103:2580–2592
De Souza MJ, Nair S, Loka Bharathi PA, Chandramohan D (2006) Metal and antibiotic-resistance in psychotropic bacteria from Antarctic Marine waters. Ecotoxicology 15:379–384
Engemann CA, Adams L, Knapp CW, Graham DW (2006) Disappearance of oxytetracycline resistance genes in aquatic systems. FEMS Microbiol Lett 263:176–182
Guillaume G, Verbrugge D, Chasseur-Libotte ML, Moens W, Collard JM (2000) PCR typing of tetracycline resistance determinants (Tet A-E) in Salmonella enterica serotype Hadar and in the microbial community of activated sludges from hospital and urban wastewater treatment facilities in Belgium. FEMS Microbiol Ecol 32:77–85
Hardwick SA, Stokes HW, Findlay S, Taylor M, Gillings MR (2008) Quantification of class 1 integron abundance in natural environments using real-time quantitative PCR. FEMS Microbiol Lett 278:207–212
Jacobs L, Chenia HY (2007) Characterization of integrons and tetracycline resistance determinants in Aeromonas spp. isolated from South African aquaculture systems. Int J Food Microbiol 114:295–306
Lachmayr KL, Kerkhof LJ, DiRienzo AG, Cavanaugh CM, Ford TE (2009) Quantifying nonspecific TEMβ-lactamase (blaTEM) genes in a wastewater stream. Appl Environ Microbiol 75:203–211
Liang DW, Zhang T, Fang HHP (2008) Real-time quantifications of dominant anaerobes in an upflow reactor by polymerase chain reaction using a TaqMan probe. Water Sci Technol 57:1851–1855
Macauley JJ, Adams CD, Mormile MR (2007) Diversity of tet resistance genes in tetracycline resistant bacteria isolated from a swine lagoon with low antibiotic impact. Can J Microbiol 53:1307–1315
Mackie RI, Koike S, Krapac I, Chee-Sanford J, Maxwell S, Aminov RI (2006) Tetracycline residues and tetracycline resistance genes in groundwater impacted by swine production facilities. Anim Biotechnol 17:157–176
Mazel D (2006) Integrons: agents of bacterial evolution. Nat Rev Microbiol 4:608–620
Mazel D, Dychinco B, Webb VA, Davies J (2000) Antibiotic resistance in the ECOR collection, integrons and identification of a novel aad gene. Antimicrob Agents Chemother 44:1568–1574
Moura A, Henriques I, Ribeiro R, Correia A (2007) Prevalence and characterization of integrons from bacteria isolated from a slaughterhouse wastewater treatment plant. J Antimicrob Chemother 60:1243–1250
Mukherjee S, Chakraborty R (2006) Incidence of class 1 integrons in multiple antibiotic-resistant Gram-negative copiotrophic bacteria from the River Torsa in India. Res Microbiol 157:220–226
Ng LK, Martin I, Alfa M, Mulvey M (2001) Multiplex PCR for the detection of tetracycline resistant genes. Mol Cell Probe 15:209–215
Park S, Choi K (2008) Hazard assessment of commonly used agricultural antibiotics on aquatic ecosystems. Ecotoxicology 17:526–538
Pei RT, Kim SC, Carlson KH, Pruden A (2006) Effect of river landscape on the sediment concentrations of antibiotics and corresponding antibiotic resistance genes (ARG). Water Res 40:2427–2435
Ploy MC, Lambert T, Couty JP, Denis F (2000) Integrons: an antibiotic resistance gene capture and expression system. Clin Chem Lab Med 38:483–487
Poppe C, Martin L, Muckle A, Archambault M, McEwen S, Weir E (2006) Characterization of antimicrobial resistance of Salmonella Newport isolated from animals, the environment, and animal food products in Canada. Can J Vet Res 70:105–114
Richards SM, Cole SE (2006) A toxicity and hazard assessment of fourteen pharmaceuticals to Xenopus laevis larvae. Ecotoxicology 15:647–656
Roberts MC (2002) Resistance to tetracycline, macrolide-lincosamide-streptogramin, trimethoprim, and sulfonamide drug classes. Mol Biotechnol 20:261–283
Salyers AA, Amabile-Cuevas CF (1997) Why are antibiotic resistance genes so resistant to elimination? Antimicrob Agents Chemother 41:2321–2325
Schmidt AS, Bruun MS, Dalsgaard I, Larsen JL (2001) Incidence, distribution, and spread of tetracycline resistance determinants and integron-associated antibiotic resistance genes among motile aeromonads from a fish farming environment. Appl Environ Microbiol 67:5675–5682
Szczepanowski R, Krahn I, Linke B, Goesmann A, Pühler A, Schlüter A (2004) Antibiotic multiresistance plasmid pRSB101 isolated from a wastewater treatment plant is related to plasmids residing in phytopathogenic bacteria and carries eight different resistance determinants including a multidrug transport system. Microbiology-SGM 150:3613–3630
Taviani E, Ceccarelli D, Lazaro N, Bani S, Cappuccinelli P, Colwell RR, Colombo MM (2008) Environmental Vibrio spp., isolated in Mozambique, contain a polymorphic group of integrative conjugative elements and class 1 integrons. FEMS Microbiol Ecol 64:45–54
Wang XB (2005) The trend and situation of the consumption of anti-infective drugs in 22 hospitals in Nanjing during 2001–2003. Chinese J Pharmacoepidemiol 14:34–36
Wang YL, Leung PC, Qian PY, Gu JD (2006) Antibiotic resistance and plasmid profile of environmental isolates of Vibrio species from Mai Po Nature Reserve, Hong Kong. Ecotoxicology 15:371–378
White PA, MacIver CJ, Rawlinson WD (2001) Integrons and gene cassettes in the Enterobacteriaceae. Antimicrob Agents Chemother 45:2658–2661
Yin HB, Fan CX, Ding SM, Zhang L, Zhong JC (2008) Geochemistry of iron, sulfur and related heavy metals in metal-polluted Taihu Lake sediments. Pedosphere 18:564–573
Yu ZT, Michel FC Jr, Hansen G, Wittum T, Morrison M (2005) Development and application of real-time PCR assays for quantification of genes encoding tetracycline resistance. Appl Environ Microbiol 71:6926–6933
Zhang T, Fang HHP (2006) Applications of real-time polymerase chain reaction for quantification of microorganisms in environmental samples. Appl Microbiol Biotechnol 70:281–289
Zhang T, Zhang M, Zhang XX, Fang HHP (2009a) Tetracycline resistance genes and tetracycline resistant lactose-fermenting Enterobacteriaceae in activated sludge of sewage treatment plants. Environ Sci Technol 43:3455–3460
Zhang XX, Zhang T, Fang HHP (2009b) Antibiotic resistance genes in water environment. Appl Microbiol Biotechnol 82:397–414
Zhang XX, Zhang T, Zhang M, Fang HHP, Cheng SP (2009c) Characterization and quantification of class 1 integrons and associated gene cassettes in sewage treatment plants. Appl Microbiol Biotechnol 82:1169–1177
Acknowledgments
This study was financially supported by the National Basic Research Program of China (973 program) (no. 2008CB418102), Hong Kong Research Grants Council (no. HKU 7195/06E) and International Foundation for Science (no. W/4215-1). The authors would like to thank Beihekou Tap Water Plant of Nanjing (Jiangsu, China) for the technological support and manipulation management of the biofilters treating Yangtze River source water.
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Zhang, X., Wu, B., Zhang, Y. et al. Class 1 integronase gene and tetracycline resistance genes tetA and tetC in different water environments of Jiangsu Province, China. Ecotoxicology 18, 652–660 (2009). https://doi.org/10.1007/s10646-009-0332-3
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DOI: https://doi.org/10.1007/s10646-009-0332-3