Abstract
Shellfish-growing areas in marine environments are affected by pollutants that mainly originate from land, including streams, domestic wastewater, and the effluents of wastewater treatment plants (WWTPs), which may function as reservoirs of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs). The objective of this study was to identify the occurrence and distribution of antibiotic resistance at five oyster sampling sites and 11 major inland pollution sources in the drainage basin of Kamak Bay, Republic of Korea. Culture-based methods were used to estimate the diversity and abundance of antibiotic-resistant Escherichia coli strains isolated from oysters and major inland pollution sources. The percentages of ARB and multiple antibiotic resistance index values were significantly high in discharge water from small fishing villages without WWTPs. However, the percentages of antibiotic-resistant E. coli isolates from oysters were low, as there was no impact from major inland pollutants. Fourteen ARGs were also quantified from oysters and major inland pollution sources. Although most ARGs except for quinolones were widely distributed in domestic wastewater discharge and effluent from WWTPs, macrolide resistance genes (ermB and msrA) were detected mainly from oysters in Kamak Bay. This study will aid in tracking the sources of antibiotic contamination in shellfish to determine the correlation between shellfish and inland pollution sources.
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References
Abia, A. L. K., Ubomba-Jaswa, E., & Momba, M. N. B. (2015). Impact of seasonal variation on Escherichia coli concentrations in the riverbed sediments in the Apies River, South Africa. Science of the Total Environment, 537, 462–469. https://doi.org/10.1016/j.scitotenv.2015.07.132
Alexander, J., Bollmann, A., Seitz, W., & Schwartz, T. (2015). Microbiological characterization of aquatic microbiomes targeting taxonomical marker genes and antibiotic resistance genes of opportunistic bacteria. Science of the Total Environment, 512–513, 316–325. https://doi.org/10.1016/j.scitotenv.2015.01.046
APHA (American Public Health Association) (1970). Recommended procedures for the examination of seawater and shellfish. Fourth ed. American Public Health Association, Washington D.C., U.S. pp. 1–47.
Auerbach, E. A., Seyfried, E. E., & McMahon, K. D. (2007). Tetracycline resistance genes in activated sludge wastewater treatment plants. Water Research, 41(5), 1143–1151. https://doi.org/10.1016/j.watres.2006.11.045
Bean, D. C., Livermore, D. M., Papa, I., & Hall, L. M. (2005). Resistance among Escherichia coli to sulphonamides and other antimicrobials now little used in man. Journal of Antimicrobial Chemotherapy, 56(5), 962–964. https://doi.org/10.1093/jac/dki332
Ben, W., Wang, J., Cao, R., Yang, M., Zhang, Y., & Qiang, Z. (2017). Distribution of antibiotic resistance in the effluents of ten municipal wastewater treatment plants in China and the effect of treatment processes. Chemosphere, 172, 392–398. https://doi.org/10.1016/j.chemosphere.2017.01.041
Biancani, P. J., Carmichael, R. H., Daskin, J. H., Burkhardt, W., & Calci, K. R. (2012). Seasonal and spatial effects of wastewater effluent on growth, survival, and accumulation of microbial contaminants by oysters in Mobile Bay Alabama. Estuaries and Coasts, 35(1), 121–131. https://doi.org/10.1007/s12237-011-9421-7
Börjesson, S., Matussek, A., Melin, S., Löfgren, S., & Lindgren, P. E. (2010). Methicillin-resistant Staphylococcus aureus (MRSA) in municipal wastewater: an uncharted threat? Journal of Applied Microbiology, 108(4), 1244–1251. https://doi.org/10.1111/j.1365-2672.2009.04515.x
Campos, C. J., & Cachola, R. A. (2007). Faecal coliforms in bivalve harvesting areas of the Alvor lagoon (Southern Portugal): Influence of seasonal variability and urban development. Environmental Monitoring and Assessment, 133(1–3), 31–41. https://doi.org/10.1007/s10661-006-9557-2
CLSI (Clinical and Laboratory Standards Institute). (2017). Performance standards for antimicrobial susceptibility testing. CLSI document M100-S26. Clinical and Laboratory Standards Institute. Wayne, PA, U.S.A., 29–76.
Coque, T. M., Baquero, F., & Canton, R. (2008). Increasing prevalence of ESBL-producing Enterobacteriaceae in Europe. Eurosurveillance. 13(47), 19044. https://doi.org/10.18535/jmscr/v5i11.223
Dang, H., Zhang, X., Song, L., Chang, Y., & Yang, G. (2007). Molecular determination of oxytetracycline-resistant bacteria and their resistance genes from mariculture environments of China. Journal of Applied Microbiology, 103(6), 2580–2592. https://doi.org/10.1111/j.1365-2672.2007.03494.x
Dang, H., Zhao, J., Song, L., Chen, M., & Chang, Y. (2009). Molecular characterizations of chloramphenicol-and oxytetracycline-resistant bacteria and resistance genes in mariculture waters of China. Marine Pollution Bulletin, 58(7), 987–994. https://doi.org/10.1016/j.marpolbul.2009.02.016
Derolez, V., Soudant, D., Fiandrino, A., Cesmat, L., & Serais, O. (2013). Impact of weather conditions on Escherichia coli accumulation in oysters of the Thau lagoon (the Mediterranean, France). Journal of Applied Microbiology, 14(2), 516–525. https://doi.org/10.1111/jam.12040
Evanson, M., & Ambrose, R. F. (2006). Sources and growth dynamics of fecal indicator bacteria in a coastal wetland system and potential impacts to adjacent waters. Water Research, 40(3), 475–486. https://doi.org/10.1016/j.watres.2005.11.027
Gao, P., He, S., Huang, S., Li, K., Liu, Z., Xue, G., & Sun, W. (2015). Impacts of coexisting antibiotics, antibacterial residues, and heavy metals on the occurrence of erythromycin resistance genes in urban wastewater. Applied Microbiology and Biotechnology, 99(9), 3971–3980. https://doi.org/10.1007/s00253-015-6404-9
Geary, P. M., & Davies, C. M. (2003). Bacterial source tracking and shellfish contamination in a coastal catchment. Water Science and Technology, 47(7–8), 95–100. https://doi.org/10.2166/wst.2003.0676
Glasoe, S., & Christy, A. (2004). Coastal urbanization and microbial contamination of shellfish growing areas. Puget Sound Action Team Publication (PSAT). Olympia, WA, 04–09.
Gullberg, E., Cao, S., Berg, O. G., Ilbäck, C., Sandegren, L., Hughes, D., & Andersson, D. I. (2011). Selection of resistant bacteria at very low antibiotic concentrations. PLoS Pathogens, 7(7), e1002158. https://doi.org/10.1371/journal.ppat.1002158
Hall, B. G., & Barlow, M. (2004). Evolution of the serine β-lactamases: Past, present and future. Drug Resistance Updates, 7(2), 111–123. https://doi.org/10.1007/978-1-4614-1400-1_12
Hammerum, A. M., Sandvang, D., Andersen, S. R., Seyfarth, A. M., Porsbo, L. J., Frimodt-Møller, N., & Heuer, O. E. (2006). Detection of sul1, sul2 and sul3 in sulphonamide resistant Escherichia coli isolates obtained from healthy humans, pork and pigs in Denmark. International Journal of Food Microbiology, 106(2), 235–237. https://doi.org/10.1016/j.ijfoodmicro.2005.06.023
Hastings, P. J., Rosenberg, S. M., & Slack, A. (2004). Antibiotic-induced lateral transfer of antibiotic resistance. Trends in microbiology, 12(9), 401–404. https://doi.org/10.1016/j.tim.2004.07.003
Hernández, A., Sanchez, M. B., & Martínez, J. L. (2011). Quinolone resistance: Much more than predicted. Frontiers in Microbiology, 2(22), 1–6. https://doi.org/10.3389/fmicb.2011.00022
Huang, Y., Zhang, L., Tiu, L., & Wang, H. H. (2015a). Characterization of antibiotic resistance in commensal bacteria from an aquaculture ecosystem. Frontiers in Microbiology, 6(914), 1–7. https://doi.org/10.3389/fmicb.2015.00914
Huang, X., Liu, C., Li, K., Su, J., Zhu, G., & Liu, L. (2015b). Performance of vertical up-flow constructed wetlands on swine wastewater containing tetracyclines and tet genes. Water Research, 70, 109–117. https://doi.org/10.1016/j.watres.2014.11.048
Huang, H., Chen, Y., Zheng, X., Su, Y., Wan, R., & Yang, S. (2016). Distribution of tetracycline resistance genes in anaerobic treatment of waste sludge: The role of pH in regulating tetracycline resistant bacteria and horizontal gene transfer. Bioresource Technology, 218, 1284–1289. https://doi.org/10.1016/j.biortech.2016.07.097
Hummel, A., Holzapfel, W. H., & Franz, C. M. (2007). Characterisation and transfer of antibiotic resistance genes from enterococci isolated from food. Systematic and Applied Microbiology, 30(1), 1–7. https://doi.org/10.1016/j.syapm.2006.02.004
Jacoby, G., & Bush, K. (2005). β‐Lactam Resistance in the 21st Century. Frontiers in antimicrobial resistance: a tribute to Stuart B. Levy, 53–65. https://doi.org/10.1128/9781555817572.ch5
Jeong, S. H., Shin, S. B., Lee, J. H., Kwon, J. Y., Lee, H. C., Kim, S. J., & Ha, K. S. (2020). Level of contamination in the feces of several species at major inland pollution sources in the drainage basin of Yeoja Bay, Republic of Korea. Environmental Monitoring and Assessment, 192(3), 170. https://doi.org/10.1007/s10661-020-8131-7
Jung, W. K., Koo, H. C., Kim, K. W., Shin, S., Kim, S. H., & Park, Y. H. (2008). Antibacterial activity and mechanism of action of the silver ion in Staphylococcus aureus and Escherichia coli. Applied and Environmental Microbiology, 74(7), 2171–2178. https://doi.org/10.3389/fmicb.2020.582349
Jung, Y. J., Park, Y. C., Lee, K. J., Kim, M. S., Go, K. R., Park, S. G., Kwon, S. J., Yang, J. H., & Mok, J. S. (2017). Spatial and seasonal variation of pollution sources in proximity of the Jaranman-Saryangdo area in Korea. Marine Pollution Bulletin, 115(1–2), 369–375. https://doi.org/10.1016/j.marpolbul.2016.12.003
Kümmerer, K. (2009a). Antibiotics in the aquatic environment—A review—Part I. Chemosphere, 75(4), 417–434. https://doi.org/10.1016/j.chemosphere.2008.11.086
Kümmerer, K. (2009b). Antibiotics in the aquatic environment—A review—Part II. Chemosphere, 75(4), 435–441. https://doi.org/10.1016/j.chemosphere.2008.12.006
Kumar, A., & Pal, D. (2018). Antibiotic resistance and wastewater: Correlation, impact and critical human health challenges. Journal of Environmental Chemical Engineering, 6(1), 52–58. https://doi.org/10.1016/j.jece.2017.11.059
Kwon, J. Y., Kwon, S. J., Yang, J. H., Mok, J. S., Jeong, S. H., Ha, K. S., Lee, H. J., & Jung, Y. J. (2019). Antimicrobial resistance of Escherichia coli isolated from oysters Crassostrea gigas and major inland pollution sources in the Jaranman-Saryangdo area in Korea. Korean Journal of Fisheries and Aquatic Sciences, 52(6), 605–616. https://doi.org/10.5657/KFAS.2019.0605
Lachmayr, K. L., Kerkhof, L. J., DiRienzo, A. G., Cavanaugh, C. M., & Ford, T. E. (2009). Quantifying nonspecific TEM β-lactamase (blaTEM) genes in a wastewater stream. Applied and Environmental Microbiology, 75(1), 203–211. https://doi.org/10.1128/aem.01254-08
Lipp, E. K., Kurz, R., Vincent, R., Rodriguez-Palacios, C., Farrah, S. R., & Rose, J. B. (2001). The effects of seasonal variability and weather on microbial fecal pollution and enteric pathogens in a subtropical estuary. Estuaries, 24(2), 266–276. https://doi.org/10.2307/1352950
Marti, E., Jofre, J., & Balcazar, J. L. (2013). Prevalence of antibiotic resistance genes and bacterial community composition in a river influenced by a wastewater treatment plant. PLoS One, 8(10), e78906. https://doi.org/10.1371/journal.pone.0078906
Marti, E., Variatza, E., & Balcazar, J. L. (2014). The role of aquatic ecosystems as reservoirs of antibiotic resistance. Trends in Microbiology, 22(1), 36–41. https://doi.org/10.1016/j.tim.2013.11.001
Matthew, M., Hedges, R. W., & Smith, J. T. (1979). Types of beta-lactamase determined by plasmids in gram-negative bacteria. Journal of Bacteriology, 138(3), 657–662. https://doi.org/10.1128/jb.138.3.657-662.1979
Mok, J. S., Lee, K. J., Kim, P. H., Lee, T. S., Lee, H. J., Jung, Y. J., & Kim, J. H. (2016). Bacteriological quality evaluation of seawater and oysters from the Jaranman-Saryangdo area, a designated shellfish growing area in Korea: Impact of inland pollution sources. Marine Pollution Bulletin, 108(1–2), 147–154. https://doi.org/10.1016/j.marpolbul.2016.04.036
Mok, J. S., Ryu, A., Kwon, J. Y., Kim, B., & Park, K. (2019). Distribution of Vibrio species isolated from bivalves and bivalve culture environments along the Gyeongnam coast in Korea Virulence and antimicrobial resistance of Vibrio parahaemolyticus isolates. Food Control, 106, 106697. https://doi.org/10.1016/j.foodcont.2019.06.023
Nijssen, S., Florijn, A., Bonten, M. J. M., Schmitz, F. J., Verhoef, J., & Fluit, A. C. (2004). Beta-lactam susceptibilities and prevalence of ESBL-producing isolates among more than 5,000 European Enterobacteriaceae isolates. International Journal of Antimicrobial Agents, 24(6), 585–591. https://doi.org/10.1016/j.ijantimicag.2004.08.008
Oksanen, J., Blanchet, F. G., Kindt, R., Legendre, P., O’hara, R. B., Simpson, G. L., Stevens, M. H. H., Wagner, H. (2010). Vegan: community ecology package. R package version 1.17-8. http://CRAN.Rproject.org/package=vegan
Ozaktas, T., Taskin, B., & Gozen, A. G. (2012). High level multiple antibiotic resistance among fish surface associated bacterial populations in non-aquaculture freshwater environment. Water Research, 46(19), 6382–6390. https://doi.org/10.1016/j.watres.2012.09.010
Portillo, A., Ruiz-Larrea, F., Zarazaga, M., Alonso, A., Martinez, J. L., & Torres, C. (2000). Macrolide resistance genes in Enterococcus spp. Antimicrobial Agents and Chemotherapy, 44(4), 967–971. https://doi.org/10.1128/aac.44.4.967-971.2000
Poirel, L., Rodriguez-Martinez, J. M., Mammeri, H., Liard, A., & Nordmann, P. (2005). Origin of plasmid-mediated quinolone resistance determinant QnrA. Antimicrobial Agents and Chemotherapy, 49(8), 3523–3525. https://doi.org/10.1128/aac.49.8.3523-3525.2005
Poirel, L., Leviandier, C., & Nordmann, P. (2006). Prevalence and genetic analysis of plasmid-mediated quinolone resistance determinants QnrA and QnrS in Enterobacteriaceae isolates from a French university hospital. Antimicrobial Agents and Chemotherapy, 50(12), 3992–3997. https://doi.org/10.1128/aac.00597-06
Proia, L., von Schiller, D., Sànchez-Melsió, A., Sabater, S., Borrego, C. M., Rodríguez-Mozaz, S., & Balcázar, J. L. (2016). Occurrence and persistence of antibiotic resistance genes in river biofilms after wastewater inputs in small rivers. Environmental Pollution, 210, 121–128. https://doi.org/10.1016/j.envpol.2015.11.035
Rafraf, I. D., Lekunberri, I., Sànchez-Melsió, A., Aouni, M., Borrego, C. M., & Balcázar, J. L. (2016). Abundance of antibiotic resistance genes in five municipal wastewater treatment plants in the Monastir Governorate, Tunisia. Environmental Pollution, 219, 353–358. https://doi.org/10.1016/j.envpol.2016.10.062
Rieke, E. L., Moorman, T. B., Douglass, E. L., & Soupir, M. L. (2018). Seasonal variation of macrolide resistance gene abundances in the South Fork Iowa River Watershed. Science of the Total Environment, 610, 1173–1179. https://doi.org/10.1016/j.scitotenv.2017.08.116
Rizzo, L., Manaia, C., Merlin, C., Schwartz, T., Dagot, C., Ploy, M. C., Michael, I., & Fatta-Kassinos, D. (2013). Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: A review. Science of the Total Environment, 447, 345–360. https://doi.org/10.1016/j.scitotenv.2013.01.032
Robicsek, A., Jacoby, G. A., & Hooper, D. C. (2006). The worldwide emergence of plasmid-mediated quinolone resistance. The Lancet Infectious Diseases, 6(10), 629–640. https://doi.org/10.1016/s1473-3099(06)70599-0
Schmitz, F. J., Sadurski, R., Kray, A., Boos, M., Geisel, R., Köhrer, K., Verhoef, J., & Fluit, A. C. (2000). Prevalence of macrolide-resistance genes in Staphylococcus aureus and Enterococcus faecium isolates from 24 European university hospitals. Journal of Antimicrobial Chemotherapy, 45(6), 8910–9894. https://doi.org/10.1093/jac/45.6.891
Seyfried, E. E., Newton, R. J., Rubert, K. F., Pedersen, J. A., & McMahon, K. D. (2010). Occurrence of tetracycline resistance genes in aquaculture facilities with varying use of oxytetracycline. Microbial Ecology, 59(4), 799–807. https://doi.org/10.1007/s00248-009-9624-7
Shah, S. Q., Cabello, F. C., L’Abée-Lund, T. M., Tomova, A., Godfrey, H. P., Buschmann, A. H., & Sørum, H. (2014). Antimicrobial resistance and antimicrobial resistance genes in marine bacteria from salmon aquaculture and non-aquaculture sites. Environmental Microbiology, 16(5), 1310–1320. https://doi.org/10.1111/1462-2920.12421
Sköld, O. (2000). Sulfonamide resistance: Mechanisms and trends. Drug Resistance Updates, 3(3), 155–160. https://doi.org/10.1054/drup.2000.0146
Stoll, C., Sidhu, J. P., Tiehm, A., & Toze, S. (2012). Prevalence of clinically relevant antibiotic resistance genes in surface water samples collected from Germany and Australia. Environmental Science and Technology, 46(17), 9716–9726. https://doi.org/10.1021/es302020s
Suzuki, S., Pruden, A., Virta, M., & Zhang, T. (2017). Antibiotic resistance in aquatic systems. Frontiers in Microbiology, 8(14), 1–3. https://doi.org/10.3389/978-2-88945-131-9
Taviani, E., Ceccarelli, D., Lazaro, N., Bani, S., Cappuccinelli, P., Colwell, R. R., & Colombo, M. M. (2008). Environmental Vibrio spp., isolated in Mozambique, contain a polymorphic group of integrative conjugative elements and class 1 integrons. FEMS Microbiology Ecology, 64(1), 45–54. https://doi.org/10.1111/j.1574-6941.2008.00455.x.
Taylor, N. G., Verner-Jeffreys, D. W., & Baker-Austin, C. (2011). Aquatic systems: Maintaining, mixing and mobilising antimicrobial resistance? Trends in Ecology and Evolution, 26(6), 278–284. https://doi.org/10.1016/j.tree.2011.03.004
Thiele-Bruhn, S. (2003). Pharmaceutical antibiotic compounds in soils—A review. Journal of Plant Nutrition and Soil Science, 166(2), 145–167. https://doi.org/10.1002/jpln.200390023
US FDA. (2015). National Shellfish Sanitation Program guide for the control of molluscan shellfish 2015 revisions. Available online: https://www.fda.gov/Food/GuidanceRegulation/FederalStateFoodPrograms/default.htm. (Accessed 10 August 2018).
US FDA. (2017). Enumeration of Escherichia coli and the Coliform Bacteria: Chapter 4. US FDA Bacteriological Analytical Manual online. Available from: http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/UCM070149. (Accessed 10 August 2018).
Wang, R. X., Wang, A., & Wang, J. Y. (2014). Antibiotic resistance monitoring in heterotrophic bacteria from anthropogenic-polluted seawater and the intestines of oyster Crassostrea hongkongensis. Ecotoxicology and Environmental Safety, 109, 27–31. https://doi.org/10.1016/j.ecoenv.2014.07.028
Warnes, M. G. R., Bolker, B., Bonebakker, L., Gentlman, R., & Huber, W. (2016). Package ‘gplots’. Various R programming tools for plotting data.
Wen, Q., Yang, L., Duan, R., & Chen, Z. (2016). Monitoring and evaluation of antibiotic resistance genes in four municipal wastewater treatment plants in Harbin, Northeast China. Environmental Pollution, 212, 34–40. https://doi.org/10.1016/j.envpol.2016.01.043
Xi, C., Zhang, Y., Marrs, C. F., Ye, W., Simon, C., Foxman, B., & Nriagu, J. (2009). Prevalence of antibiotic resistance in drinking water treatment and distribution systems. Applied and Environmental Microbiology, 75(17), 5714–5718. https://doi.org/10.5353/th_b5784873
Yang, Y., Zhang, T., Zhang, X. X., Liang, D. W., Zhang, M., Gao, D. W., Zhu, H. G., Huang, Q. G., & Fang, H. H. P. (2012). Quantification and characterization of β-lactam resistance genes in 15 sewage treatment plants from East Asia and North America. Applied Microbiology and Biotechnology, 95(5), 1351–1358. https://doi.org/10.1007/s00253-011-3810-5
Zhang, H., Liu, P., Feng, Y., & Yang, F. (2013). Fate of antibiotics during wastewater treatment and antibiotic distribution in the effluent-receiving waters of the Yellow Sea, northern China. Marine pollution bulletin, 73(1), 282–290. https://doi.org/10.1016/j.marpolbul.2013.05.007
Zhang, X. X., Zhang, T., & Fang, H. H. (2009). Antibiotic resistance genes in water environment. Applied Microbiology and Biotechnology, 82(3), 397–414. https://doi.org/10.1016/b978-0-12-801214-7.00016-8
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Jeong, S.H., Kwon, J.Y., Shin, S.B. et al. Antibiotic resistance in shellfish and major inland pollution sources in the drainage basin of Kamak Bay, Republic of Korea. Environ Monit Assess 193, 471 (2021). https://doi.org/10.1007/s10661-021-09201-z
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DOI: https://doi.org/10.1007/s10661-021-09201-z