Metagenomic analysis reveals wastewater treatment plants as hotspots of antibiotic resistance genes and mobile genetic elements

Water Research

Volumn 123, Issue , 2017, Pages 468-478

  Guo, Jianhua ^a   Li, Jie ^b   Chen, Hui ^a   Bond, Philip L ^a   Yuan, Zhiguo ^a  

^a UNIVERSITY OF QUEENSLAND   (Australia)

^b BGI SHENZHEN   (China)

Author keywords

Antibiotic resistance genes (ARGs); Antibiotic resistant bacteria (ARB); Antibiotics; Biological wastewater treatment; Metagenomics; Mobile genetic elements (MGEs)

Indexed keywords

ACTIVATED SLUDGE PROCESS; ANAEROBIC DIGESTION; ANTIBIOTICS; BACTERIA; BIOLOGICAL WATER TREATMENT; DNA; GENE TRANSFER; GENES; RECLAMATION; SEWAGE PUMPING PLANTS; WASTEWATER TREATMENT; WATER TREATMENT PLANTS;

ANTIBIOTIC RESISTANCE GENES; ANTIBIOTIC-RESISTANT BACTERIA; BIOLOGICAL WASTE WATER TREATMENT; GENETIC ELEMENTS; METAGENOMICS;

SEWAGE TREATMENT PLANTS;

AMIKACIN; AMINOGLYCOSIDE ANTIBIOTIC AGENT; BETA LACTAM ANTIBIOTIC; CHLORAMPHENICOL; DIBEKACIN; ISEPAMICIN; LINCOSAMIDE; MACROLIDE; NETILMICIN; POLYPEPTIDE ANTIBIOTIC AGENT; QUINOLINE DERIVED ANTIINFECTIVE AGENT; SISOMICIN; SPECTINOMYCIN; STREPTOGRAMIN DERIVATIVE; STREPTOMYCIN; SULFONAMIDE; TETRACYCLINE DERIVATIVE; TOBRAMYCIN; TRIMETHOPRIM; ANTIINFECTIVE AGENT; WASTE WATER;

ACTIVATED SLUDGE; ANAEROBIC DIGESTION; ANTIBIOTIC RESISTANCE; ANTIBIOTICS; BACTERIUM; GENE; GENE EXPRESSION; GENE TRANSFER; GENETIC ANALYSIS; GENOMICS; MICROBIAL COMMUNITY; WASTEWATER; WASTEWATER TREATMENT; WASTEWATER TREATMENT PLANT;

ACTINOBACTERIA; ACTIVATED SLUDGE; ANAEROBIC DIGESTION; ANTIBIOTIC RESISTANCE; ANTIBIOTIC RESISTANCE GENE; ARTICLE; BACTERIAL GENOME; BACTEROIDETES; CLOACIMONETES; CLOSTRIDIUM; COMMUNITY STRUCTURE; DNA SEQUENCE; EURYARCHAEOTA; FIRMICUTES; GENE INSERTION SEQUENCE; GENE STRUCTURE; GENE TRANSFER; GENETIC VARIABILITY; HIGH THROUGHPUT SEQUENCING; INTEGRON; METAGENOMICS; MICROBIAL COMMUNITY; MOBILE GENETIC ELEMENT; MOLECULAR PHYLOGENY; NITROSOMONAS; NITROSPIRAE; NONHUMAN; OPEN READING FRAME; PLASMID; PRIORITY JOURNAL; PROTEOBACTERIA; TAXONOMY; TRANSPOSON; WASTE WATER MANAGEMENT; WASTE WATER TREATMENT PLANT; BACTERIAL GENE; BACTERIUM; GENETICS; HIGH THROUGHPUT SCREENING; PHYLOGENY; WASTE WATER;

CLOSTRIDIUM; NITROSOMONAS;

ANTI-BACTERIAL AGENTS; BACTERIA; DRUG RESISTANCE, MICROBIAL; GENES, BACTERIAL; HIGH-THROUGHPUT SCREENING ASSAYS; PHYLOGENY; WASTE WATER;

EID: 85021760467     PISSN: 00431354     EISSN: 18792448     Source Type: Journal    
DOI: 10.1016/j.watres.2017.07.002     Document Type: Article

References (81)

1. Alonso, A., Sánchez, P., Martínez, J.L., Environmental selection of antibiotic resistance genes. Environ. Microbiol. 3:1 (2001), 1–9.
2. Aminov, R.I., The role of antibiotics and antibiotic resistance in nature. Environ. Microbiol. 11:12 (2009), 2970–2988.
3. Ashbolt, N.J., Amezquita, A., Backhaus, T., Borriello, P., Brandt, K.K., Collignon, P., Coors, A., Finley, R., Gaze, W.H., Heberer, T., Lawrence, J.R., Larsson, D.G.J., McEwen, S.A., Ryan, J.J., Schonfeld, J., Silley, P., Snape, J.R., Van den Eede, C., Topp, E., Human health risk assessment (HHRA) for environmental development and transfer of antibiotic resistance. Environ. Health Perspect. 121:9 (2013), 993–1001.
4. Bengtsson-Palme, J., Hammaren, R., Pal, C., Ostman, M., Bjorlenius, B., Flach, C.F., Fick, J., Kristiansson, E., Tysklind, M., Larsson, D.G.J., Elucidating selection processes for antibiotic resistance in sewage treatment plants using metagenomics. Sci. Total Environ. 572 (2016), 697–712.
5. Bennett, P.M., Plasmid encoded antibiotic resistance: acquisition and transfer of antibiotic resistance genes in bacteria. Br. J. Pharmacol. 153:1 (2008), S347–S357.
6. Berendonk, T.U., Manaia, C.M., Merlin, C., Fatta-Kassinos, D., Cytryn, E., Walsh, F., Buergmann, H., Sorum, H., Norstrom, M., Pons, M.-N., Kreuzinger, N., Huovinen, P., Stefani, S., Schwartz, T., Kisand, V., Baquero, F., Luis Martinez, J., Tackling antibiotic resistance: the environmental framework. Nat. Rev. Microbiol. 13:5 (2015), 310–317.
7. Calero-Caceres, W., Melgarejo, A., Colomer-Lluch, M., Stoll, C., Lucena, F., Jofre, J., Muniesa, M., Sludge as a potential important source of antibiotic resistance genes in both the bacterial and bacteriophage fractions. Environ. Sci. Technol. 48:13 (2014), 7602–7611.
8. Carey, D.E., Zitomer, D.H., Hristova, K.R., Kappell, A.D., McNamara, P.J., Triclocarban influences antibiotic resistance and alters anaerobic digester microbial community structure. Environ. Sci. Technol. 50:1 (2016), 126–134.
9. Chen, B., Yang, Y., Liang, X., Yu, K., Zhang, T., Li, X., Metagenomic profiles of antibiotic resistance genes (ARGs) between human impacted estuary and deep ocean sediments. Environ. Sci. Technol. 47:22 (2013), 12753–12760.
10. Chen, B., Yuan, K., Chen, X., Yang, Y., Zhang, T., Wang, Y., Luan, T., Zou, S., Li, X., Metagenomic analysis revealing antibiotic resistance genes (ARGs) and their genetic compartments in the Tibetan environment. Environ. Sci. Technol. 50:13 (2016), 6670–6679.
11. Christgen, B., Yang, Y., Ahammad, S.Z., Li, B., Rodriquez, D.C., Zhang, T., Graham, D.W., Metagenomics shows that low-energy anaerobic−aerobic treatment reactors reduce antibiotic resistance gene levels from domestic wastewater. Environ. Sci. Technol. 49:4 (2015), 2577–2584.
12. Diehl, D.L., LaPara, T.M., Effect of temperature on the fate of genes encoding tetracycline resistance and the integrase of Class 1 Integrons within anaerobic and aerobic digesters treating municipal wastewater solids. Environ. Sci. Technol. 44:23 (2010), 9128–9133.
13. Filee, J., Siguier, P., Chandler, M., Insertion sequence diversity in Archaea. Microbiol. Mol. Biol. Rev. 71:1 (2007), 121–157.
14. Ghosh, S., Ramsden, S.J., LaPara, T.M., The role of anaerobic digestion in controlling the release of tetracycline resistance genes and class 1 integrons from municipal wastewater treatment plants. Appl. Microbiol. Biotechnol. 84:4 (2009), 791–796.
15. Guo, J., Ni, B.-J., Han, X., Chen, X., Bond, P., Peng, Y., Yuan, Z., Unraveling microbial structure and diversity of activated sludge in a full-scale simultaneous nitrogen and phosphorus removal plant using metagenomic sequencing. Enzyme Microb. Technol. 102 (2017), 16–25.
16. Gupta, S.K., Padmanabhan, B.R., Diene, S.M., Lopez-Rojas, R., Kempf, M., Landraud, L., Rolain, J.-M., ARG-ANNOT, a new bioinformatic tool to discover antibiotic resistance genes in bacterial genomes. Antimicrob. Agents Chemother. 58:1 (2014), 212–220.
17. Huson, D.H., Weber, N., Microbial community analysis using MEGAN. DeLong, E.F., (eds.) Microbial Metagenomics, Metatranscriptomics, and Metaproteomics, 2013, 465–485.
18. Ibrahim, M.E., Bilal, N.E., Hamid, M.E., Increased multi-drug resistant Escherichia coli from hospitals in Khartoum state. Sudan. Afr. Health Sci. 12:3 (2012), 368–375.
19. Joy, S.R., Bartelt-Hunt, S.L., Snow, D.D., Gilley, J.E., Woodbury, B.L., Parker, D.B., Marx, D.B., Li, X., Fate and transport of antimicrobials and antimicrobial resistance genes in soil and runoff following land application of swine manure slurry. Environ. Sci. Technol. 47:21 (2013), 12081–12088.
20. Junjie, Q., Ruiqiang, L., Raes, J., Arumugam, M., Burgdorf, K.S., Manichanh, C., Nielsen, T., Pons, N., Levenez, F., Yamada, T., Mende, D.R., Junhua, L., Junming, X., Shaochuan, L., Dongfang, L., Jianjun, C., Bo, W., Huiqing, L., Huisong, Z., Yinlong, X., Tap, J., Lepage, P., Bertalan, M., Batto, J.M., Hansen, T., Paslier, D.l., Linneberg, A., Nielsen, H.B., Pelletier, E., Renault, P., Sicheritz-Ponten, T., Turner, K., Hongmei, Z., Chang, Y., Shengting, L., Min, J., Yan, Z., Yingrui, L., Xiuqing, Z., Songgang, L., Nan, Q., Huanming, Y., Jian, W., Brunak, S., Dore, J., Guarner, F., Kristiansen, K., Pedersen, O., Parkhill, J., Weissenbach, J., Bork, P., Ehrlich, S.D., Jun, W., Meta, H.I.T.C., A human gut microbial gene catalogue established by metagenomic sequencing. Nature 464:7285 (2010), 59–65 UK.
21. Kanehisa, M., Goto, S., Hattori, M., Aoki-Kinoshita, K.F., Itoh, M., Kawashima, S., Katayama, T., Araki, M., Hirakawa, M., From genomics to chemical genomics: new developments in KEGG. Nucleic acids. Res. 34 (2006), D354–D357.
22. Koike, S., Krapac, I.G., Oliver, H.D., Yannarell, A.C., Chee-Sanford, J.C., Aminov, R.I., Mackie, R.I., Monitoring and source tracking of tetracycline resistance genes in lagoons and groundwater adjacent to swine production facilities over a 3-year period. Appl. Environ. Microbiol. 73:15 (2007), 4813–4823.
23. Kristiansson, E., Fick, J., Janzon, A., Grabic, R., Rutgersson, C., Weijdegard, B., Soderstrom, H., Larsson, D.G.J., Pyrosequencing of antibiotic-contaminated river sediments reveals high levels of resistance and gene transfer elements. PLoS One, 6(2), 2011.
24. Krzywinski, M., Schein, J., Birol, I., Connors, J., Gascoyne, R., Horsman, D., Jones, S.J., Marra, M.A., Circos: an information aesthetic for comparative genomics. Genome Res. 19:9 (2009), 1639–1645.
25. LaPara, T.M., Burch, T.R., McNamara, P.J., Tan, D.T., Yan, M., Eichmiller, J.J., Tertiary-treated municipal wastewater is a significant point source of antibiotic resistance genes into duluth-superior harbor. Environ. Sci. Technol. 45:22 (2011), 9543–9549.
26. Lapworth, D.J., Baran, N., Stuart, M.E., Ward, R.S., Emerging organic contaminants in groundwater: a review of sources, fate and occurrence. Environ. Pollut. 163 (2012), 287–303.
27. Larson, E., Community factors in the development of antibiotic resistance. Annu. Rev. Public Health, 2007, 435–447.
28. Li, B., Yang, Y., Ma, L., Ju, F., Guo, F., Tiedje, J.M., Zhang, T., Metagenomic and network analysis reveal wide distribution and co-occurrence of environmental antibiotic resistance genes. ISME J. 9:11 (2015), 2490–2502.
29. Li, B., Zhang, X., Guo, F., Wu, W., Zhang, T., Characterization of tetracycline resistant bacterial community in saline activated sludge using batch stress incubation with high-throughput sequencing analysis. Water Res. 47:13 (2013), 4207–4216.
30. Li, D., Yang, M., Hu, J., Zhang, J., Liu, R., Gu, X., Zhang, Y., Wang, Z., Antibiotic-resistance profile in environmental bacteria isolated from penicillin production wastewater treatment plant and the receiving river. Environ. Microbiol. 11:6 (2009), 1506–1517.
31. Li, R., Li, Y., Kristiansen, K., Wang, J., SOAP: short oligonucleotide alignment program. Bioinformatics 24:5 (2008), 713–714.
32. Li, R., Yu, C., Li, Y., Lam, T.-W., Yiu, S.-M., Kristiansen, K., Wang, J., SOAP2: an improved ultrafast tool for short read alignment. Bioinformatics 25:15 (2009), 1966–1967.
33. Li, R., Zhu, H., Ruan, J., Qian, W., Fang, X., Shi, Z., Li, Y., Li, S., Shan, G., Kristiansen, K., Li, S., Yang, H., Wang, J., Wang, J., De novo assembly of human genomes with massively parallel short read sequencing. Genome Res. 20:2 (2010), 265–272.
34. Li, X., Watanabe, N., Xiao, C., Harter, T., McCowan, B., Liu, Y., Atwill, E.R., Antibiotic-resistant E. coli in surface water and groundwater in dairy operations in Northern California. Environ. Monit. Assess. 186:2 (2014), 1253–1260.
35. Liu, M., Zhang, Y., Yang, M., Tian, Z., Ren, L., Zhang, S., Abundance and distribution of tetracycline resistance genes and mobile elements in an oxytetracycline production wastewater treatment system. Environ. Sci. Technol. 46:14 (2012), 7551–7557.
36. Lupo, A., Coyne, S., Berendonk, T.U., Origin and evolution of antibiotic resistance: the common mechanisms of emergence and spread in water bodies. Front. Microbiol., 3, 2012, 18.
37. Mahillon, J., Chandler, M., Insertion sequences. Microbiol. Mol. Biol. Rev. 62:3 (1998), 725–774.
38. Mao, D., Yu, S., Rysz, M., Luo, Y., Yang, F., Li, F., Hou, J., Mu, Q., Alvarez, P.J.J., Prevalence and proliferation of antibiotic resistance genes in two municipal wastewater treatment plants. Water Res. 85 (2015), 458–466.
39. Marti, E., Variatza, E., Luis Balcazar, J., The role of aquatic ecosystems as reservoirs of antibiotic resistance. Trends Microbiol. 22:1 (2014), 36–41.
40. Miller, J.H., Novak, J.T., Knocke, W.R., Pruden, A., Survival of antibiotic resistant bacteria and horizontal gene transfer control antibiotic resistance gene content in anaerobic digesters. Front. Microbiol., 7, 2016, 263.
41. Miller, J.R., Koren, S., Sutton, G., Assembly algorithms for next-generation sequencing data. Genomics 95:6 (2010), 315–327.
42. Moura, A., Pereira, C., Henriques, I., Correia, A., Novel gene cassettes and integrons in antibiotic-resistant bacteria isolated from urban wastewaters. Res. Microbiol. 163:2 (2012), 92–100.
43. Moura, A., Soares, M., Pereira, C., Leitão, N., Henriques, I., Correia, A., INTEGRALL: a database and search engine for integrons, integrases and gene cassettes. Bioinformatics 25:8 (2009), 1096–1098.
44. Mullany, P., Functional metagenomics for the investigation of antibiotic resistance. Virulence 5:3 (2014), 443–447.
45. Munck, C., Albertsen, M., Telke, A., Ellabaan, M., Nielsen, P.H., Sommer, M.O.A., Limited dissemination of the wastewater treatment plant core resistome. Nat. Commun., 6, 2015.
46. Munir, M., Wong, K., Xagoraraki, I., Release of antibiotic resistant bacteria and genes in the effluent and biosolids of five wastewater utilities in Michigan. Water Res. 45:2 (2011), 681–693.
47. Novo, A., André, S., Viana, P., Nunes, O.C., Manaia, C.M., Antibiotic resistance, antimicrobial residues and bacterial community composition in urban wastewater. Water Res. 47:5 (2013), 1875–1887.
48. Paiva, M.C., Reis, M.P., Costa, P.S., Dias, M.F., Bleicher, L., Scholte, L.L.S., Nardi, R.M.D., Nascimento, A.M.A., Identification of new bacteria harboring qnrS and aac6 ‘-Ib/cr and mutations possibly involved in fluoroquinolone resistance in raw sewage and activated sludge samples from a full-scale WWTP. Water Res. 110 (2017), 27–37.
49. Parnanen, K., Karkman, A., Tamminen, M., Lyra, C., Hultman, J., Paulin, L., Virta, M., Evaluating the mobility potential of antibiotic resistance genes in environmental resistomes without metagenomics. Sci. Rep., 6, 2016.
50. Port, J.A., Cullen, A.C., Wallace, J.C., Smith, M.N., Faustman, E.M., Metagenomic frameworks for monitoring antibiotic resistance in aquatic environments. Environ. Health Perspect. 122:3 (2014), 222–228.
51. Port, J.A., Wallace, J.C., Griffith, W.C., Faustman, E.M., Metagenomic profiling of microbial composition and antibiotic resistance determinants in puget sound. PLoS One, 7(10), 2012.
52. Powell, S., Forslund, K., Szklarczyk, D., Trachana, K., Roth, A., Huerta-Cepas, J., Gabaldón, T., Rattei, T., Creevey, C., Kuhn, M., Jensen, L.J., von Mering, C., Bork, P., eggNOG v4.0: nested orthology inference across 3686 organisms. Nucleic Acids Res. 42:D1 (2014), D231–D239.
53. Pruden, A., Balancing water sustainability and public health goals in the face of growing concerns about antibiotic resistance. Environ. Sci. Technol. 48:1 (2014), 5–14.
54. Pruden, A., Arabi, M., Storteboom, H.N., Correlation between upstream human activities and riverine antibiotic resistance genes. Environ. Sci. Technol. 46:21 (2012), 11541–11549.
55. Rhodes, L.D., Grayson, T.H., Alexander, S.M., Strom, M.S., Description and characterization of IS994, a putative IS3 family insertion sequence from the salmon pathogen, Renibacterium salmoninarum. Gene 244:1–2 (2000), 97–107.
56. Rizzo, L., Manaia, C., Merlin, C., Schwartz, T., Dagot, C., Ploy, M.C., Michael, I., Fatta-Kassinos, D., Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Sci. Total Environ. 447 (2013), 345–360.
57. Schmieder, R., Edwards, R., Insights into antibiotic resistance through metagenomic approaches. Future Microbiol. 7:1 (2012), 73–89.
58. Shah, S.Q.A., Cabello, F.C., L'Abée-Lund, T.M., Tomova, A., Godfrey, H.P., Buschmann, A.H., Sørum, H., Antimicrobial resistance and antimicrobial resistance genes in marine bacteria from salmon aquaculture and non-aquaculture sites. Environ. Microbiol. 16:5 (2014), 1310–1320.
59. Shi, P., Jia, S., Zhang, X.-X., Zhang, T., Cheng, S., Li, A., Metagenomic insights into chlorination effects on microbial antibiotic resistance in drinking water. Water Res. 47:1 (2013), 111–120.
60. Shintani, M., Urata, M., Inoue, K., Eto, K., Habe, H., Ornori, T., Yamane, H., Nojiri, H., The Sphingomonas plasmid pCAR3 is involved in complete mineralization of carbazole. J. Bacteriol. 189:5 (2007), 2007–2020.
61. Siguier, P., Perochon, J., Lestrade, L., Mahillon, J., Chandler, M., ISfinder: the reference centre for bacterial insertion sequences. Nucleic Acids Res. 34:Suppl. 1 (2006), D32–D36.
62. Stalder, T., Barraud, O., Casellas, M., Dagot, C., Ploy, M.-C., Integron involvement in environmental spread of antibiotic resistance. Front. Microbiol., 3, 2012.
63. Sui, Q., Huang, J., Deng, S., Yu, G., Fan, Q., Occurrence and removal of pharmaceuticals, caffeine and DEET in wastewater treatment plants of Beijing, China. Water Res. 44:2 (2010), 417–426.
64. Tatusov, R.L., Galperin, M.Y., Natale, D.A., Koonin, E.V., The COG database: a tool for genome-scale analysis of protein functions and evolution. Nucleic acids. Res. 28:1 (2000), 33–36.
65. Taylor, N.G.H., Verner-Jeffreys, D.W., Baker-Austin, C., Aquatic systems: maintaining, mixing and mobilising antimicrobial resistance?. Trends Ecol. Evol. 26:6 (2011), 278–284.
66. Threedeach, S., Chiemchaisri, W., Watanabe, T., Chiemchaisri, C., Honda, R., Yamamoto, K., Antibiotic resistance of Escherichia coli in leachates from municipal solid waste landfills: comparison between semi-aerobic and anaerobic operations. Bioresour. Technol. 113 (2012), 253–258.
67. Torres-Cortés, G., Millán, V., Ramírez-Saad, H.C., Nisa-Martínez, R., Toro, N., Martínez-Abarca, F., Characterization of novel antibiotic resistance genes identified by functional metagenomics on soil samples. Environ. Microbiol. 13:4 (2011), 1101–1114.
68. Udikovic-Kolic, N., Wichmann, F., Broderick, N.A., Handelsman, J., Bloom of resident antibiotic-resistant bacteria in soil following manure fertilization. Proc. Natl. Acad. Sci. U. S. A. 111:42 (2014), 15202–15207.
69. van der Donk, C.F.M., van de Bovenkamp, J.H.B., De Brauwer, E.I.G.B., De Mol, P., Feldhoff, K.-H., Kalka-Moll, W.M., Nys, S., Thoelen, I., Trienekens, T.A.M., Stobberingh, E.E., Antimicrobial resistance and spread of multi drug resistant Escherichia coli isolates collected from nine urology services in the euregion meuse-rhine. PLoS One, 7(10), 2012, e47707.
70. van Hoek, A.H.A.M., Mevius, D., Guerra, B., Mullany, P., Roberts, A.P., Aarts, H.J.M., Acquired antibiotic resistance genes: an overview. Front. Microbiol., 2, 2011.
71. Versluis, D., D'Andrea, M.M., Garcia, J.R., Leimena, M.M., Hugenholtz, F., Zhang, J., Ozturk, B., Nylund, L., Sipkema, D., van Schaik, W., de Vos, W.M., Kleerebezem, M., Smidt, H., van Passel, M.W.J., Mining microbial metatranscriptomes for expression of antibiotic resistance genes under natural conditions. Sci. Rep., 5, 2015.
72. Wang, Z., Zhang, X.-X., Huang, K., Miao, Y., Shi, P., Liu, B., Long, C., Li, A., Metagenomic profiling of antibiotic resistance genes and mobile genetic elements in a tannery wastewater treatment plant. PLoS One, 8(10), 2013.
73. Xu, J., Xu, Y., Wang, H., Guo, C., Qiu, H., He, Y., Zhang, Y., Li, X., Meng, W., Occurrence of antibiotics and antibiotic resistance genes in a sewage treatment plant and its effluent-receiving river. Chemosphere 119:0 (2015), 1379–1385.
74. Yang, Y., Li, B., Zou, S., Fang, H.H.P., Zhang, T., Fate of antibiotic resistance genes in sewage treatment plant revealed by metagenomic approach. Water Res. 62:0 (2014), 97–106.
75. Yang, Y., Yu, K., Xia, Y., Lau, F.T.K., Tang, D.T.W., Fung, W.C., Fang, H.H.P., Zhang, T., Metagenomic analysis of sludge from full-scale anaerobic digesters operated in municipal wastewater treatment plants. Appl. Microbiol. Biot. 98:12 (2014), 5709–5718.
76. Zhang, J., Chen, M., Sui, Q., Tong, J., Jiang, C., Lu, X., Zhang, Y., Wei, Y., Impacts of addition of natural zeolite or a nitrification inhibitor on antibiotic resistance genes during sludge composting. Water Res. 91 (2016), 339–349.
77. Zhang, T., Li, B., Occurrence, transformation, and fate of antibiotics in municipal wastewater treatment plants. Crit. Rev. Environ. Sci. Technol. 41:11 (2011), 951–998.
78. Zhang, T., Yang, Y., Pruden, A., Effect of temperature on removal of antibiotic resistance genes by anaerobic digestion of activated sludge revealed by metagenomic approach. Appl. Microbiol. Biotechnol., 2015, 1–9.
79. Zhang, X.-X., Zhang, T., Fang, H.H.P., Antibiotic resistance genes in water environment. Appl. Microbiol. Biotechnol. 82:3 (2009), 397–414.
80. Zhu, W., Lomsadze, A., Borodovsky, M., Ab initio gene identification in metagenomic sequences. Nucleic acids. Res., 38(12), 2010, e132.
81. Zhu, Y.G., Johnson, T.A., Su, J.Q., Qiao, M., Guo, G.X., Stedtfeld, R.D., Hashsham, S.A., Tiedje, J.M., Diverse and abundant antibiotic resistance genes in Chinese swine farms. Proc. Natl. Acad. Sci. U. S. A. 110:9 (2013), 3435–3440.