Document Type : Research Article

Authors

1 Soil Science and Engineering department, Faculty of Agricultural Science, Guilan University, Iran

2 Soil science departement, Faculty of agricultural sience, University of Guilan, Rasht, Iran

3 Soil Science and Engineering department, Faculty of Agriculture, Bu-Ali Sina University, Iran

Abstract

Introduction
 Overuse of antibiotics and their entry into the environment is a global concern today. Much of the antibiotics taken by animals and humans are excreted unchanged and found in their feces which enter the environment through livestock waste and municipal wastewater disposal. Uncontrolled release and continuous introduction of antibiotics to the environment induced antibiotic resistance in microorganisms living in other habitats which pose a potential hazard to existing aquatic ecosystems and animals. Rivers act as the main sink for the effluents that distribute antibiotics and antibiotic resistant microorganisms in the environment. This study aimed to investigate the antibiotic resistant index (ARI) in Goharrood River. Seasonal variations and source of contamination, focusing on urban surface wastewater of Rasht, were investigated through sampling of river water and sediment in different points along the river during four seasons of a year.
Materials and Methods
The water and sediment of the river were sampled at three points along the river (focused on river course in Rasht city), in autumn, 2016 and in winter, spring and summer, 2017. The number of antibiotic resistant heterotrophic and coliform bacteria were counted via colony count method in the antibiotic supplemented 100 µg/mL Nutrient Agar and Eosine-Methylene Blue agar media respectively. Cephalexin, gentamicin, doxycycline, ciprofloxacin, and trimethoprim antibiotics were tested in this study. ARI was calculated by dividing number of bacteria colonies (heterotrophic and coliform bacteria) in plates supplemented with antibiotics to the number of colonies in control plate (without antibiotic). Escherichia coli as an indicator coliform bacterium was isolated from water and sediment samples (12 strains; 4 season and 3 sampling points) and their resistant pattern to these antibiotic was also tested by disk diffusion (Kirby-Bauer) method in Mueller-Hinton agar medium. The inhibition zone (ZOI) of E. coli growth was measured and its sensitivity/resistant was assessed based on CLSI standard protocol. The calculated ARI of heterotrophic and coliform bacteria of water and sediment of the River and the determined ZOI of E. coli isolated from water and sediment were analyzed by repeated measures of factorial arrangement in a completely randomized design format by SAS software package. Factors included sampling point at 3 levels (before entering river to the Rasht city; A, in the Rasht city; B, and after river exit from the Rasht city; C), and antibiotics at 5 levels (cephalexin, gentamicin, doxycycline, ciprofloxacin, and trimethoprim) as main plot and sampling time at 4 levels (autumn, winter, spring, and summer) as sub-plot.
Results and Discussion
 The highest ARI value of water heterotrophic bacteria was obtained to cephalexin at the sampling point C. Mean ARI of water heterotrophic bacteria to all antibiotics (regardless of type of antibiotic) at three points of A, B and C was 3.77, 4.54 and 7.53%, respectively. The highest ARI levels of heterotrophic and coliform bacteria in water were obtained in the summer season. In fact, the change of seasons and clearly the summer season controlled the ARI in water bacteria rather than the type of antibiotics. So that in this season 30.78% of water heterotrophic bacteria were resistant to ciprofloxacin antibiotic and about half (50.78) of the river water coliforms were resistant to cephalexin. Although ARI for heterotrophic and coliform bacteria was lower in sediment rather than that in water, the highest ARI levels of heterotrophic and coliform bacteria in sediment were obtained against cephalexin in autumn and winter, respectively. In general, the mean ARI in water and sediment bacteria was as follows: cephalexin > gentamicin > ciprofloxacine > doxycycline > trimethoprim. The lowest ZOI value for E. coli was obtained against cephalexin. Therefore, in all three sampling points, isolated E. coli bacteria from water and sediment were resistant to cephalexin. In the study of the sampling time and sampling point interaction, it was also seen that the lowest ZOI of E. coli was in autumn and at sampling point C. Therefore, it seems that E. coli has become resistant to antibiotics when river crosses the city.
Conclusion
 According to the results of this study, Goharrood river is contaminated with antibiotic-resistant, especially cephalexin resistant bacteria and it may distribute pollution downstream. If the river water is used in aquaculture and irrigation of downstream agriculture fields, the antibiotic resistant bacteria may be spread in the other ecosystems and finally may enter the human food chain.

Keywords

Main Subjects

  1. Akbari, Z., Shaker Khatibi, M., Mosaferi, M., Asl Rahnema Akbari, N., Shiri, Z., & Farshchian, M.R. (2016). Isolation and identification of dominant microorganisms in activated sludge of ABS effluent treatment plant and evaluation of their potential for acrylonitrile biodegradation. Journal of Urmia University of Medical Sciences 27(5): 375-383. (In Persian) URL: http://umj.umsu.ac.ir/article-1-3550-fa.html.
  2. Akiyama, T., & Savin, M.C. (2010). Populations of antibiotic-resistant coliform bacteria change rapidly in a wastewater effluent dominated stream. Science of the Total Environment 408: 6192-6201. https://doi.org/10.1016/j.scitotenv.2010.08.055.
  3. Anderl, J.N., Franklin, M.J., & Stewart, P.S. (2000). Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrobial Agents Chemotherapy 52: 3648-3663. https://doi.org/10.1128/AAC.44.7.1818-1824.2000.
  4. Bagherzadeh, A., Pirouz, B., & Sabet, A. (2018). Urban, industrial and agricultural pollutant sources on the water quality of Goharrood river in Rasht city-Gilan province, 8th International Engineering Engineering Seminar, Ahvaz. (In Persian).
  5. Bockstael, K., & Van Aerschot, A. (2009). Antimicrobial resistance in bacteria. Central European Journal of Medicine 4(2): 141-155. http://doi.org/10.2478/s11536-008-0088-9.
  6. Bonyadian, M., Moshtaghi, H., & Behroozi, P. (2017). Occurrence of verotoxigenic coli in cow feces and antimicrobial resistance of the isolates in cattle farms in Shahrekord area. Biological Journal of Microorganism 6(21): 75-84. (In Persian)
  7. Calero-Caceres, W., Mendez, J., Martín-Díaz, J., & Muniesa, M. (2017). The occurrence of antibiotic resistance genes in a Mediterranean river and their persistence in theriverbed sediment. Environmental Pollution 223: 384- https://doi.org/10.1016/j.envpol.2017.01.035.
  8. Cheng, G., Dai, M., Ahmed, S., Hao, H., Wang, X., & Yuan, Z. (2016). Antimicrobial drugs in fighting against antimicrobial resistance. Frontiers in Microbiology 7: 470. https://doi.org/10.3389/fmicb.2016.00470.
  9. CLSI, Clinical & Laboratory Standards Institute. (2014). Performance Standards for Antimicrobial Susceptibility Testing, Twenty-Fourth Informational Supplement. M100-S24. 230 p.
  10. Danner, M.C., Robertson, A., Behrends, V., & Reiss, J. (2019). Antibiotic pollution in surface fresh waters: occurrence and effects. Science of the Total Environment 664: 793-804. https://doi.org/10.1016/j.scitotenv.2019.01.406.
  11. Davin-Regli, A., Bolla, J.M., James, CE., Lavigne, J.P., Chevalier, J., Garnotel, E., & Molitor, A. (2008). Membrane permeability and regulation of drug “influx and efflux” in enterobacterial pathogens. Current Drug Targets 9(9): 750-759.
  12. Diwan, V., Hanna, N., Purohit, M., Chandran, S., Riggi, E., Parashar, V., Tamhankar, A.J., & Stålsby Lundborg, C. (2018). Seasonal variations in water-quality, antibiotic residues, resistant bacteria and antibiotic resistance genes of Escherichia coli isolates from water and sediments of the Kshipra River in Central India. International Journal of Environmental Research and Public Health 15(6): 1281. https://doi.org/10.3390/ijerph15061281.
  13. Emad,E., & Chaudhuri, M. (2011). The feasibility of using combined tioz photoeatalysis-SBR process for Antibiotic wastewater treatment. Desalination 272: 218-224. https://doi.org/10.1016/j.desal.2011.01.020.
  14. Farhangi, M., Ghorbanzadeh, N., Amini, M., & Ghovvati, S. (2021). Investigation of antibiotic resistant coliform bacteria in Zarjoub River. Iranian Journal of Soil and Water Research 52(8): 2061-2076. (In Persian). http://doi.org/10.22059/IJSWR.2021.322755.668946.
  15. Fernández, L., & Hancock, R.E.W. (2012). Adaptive and mutational resistance: role of porins and efflux pumps in drug resistance. Clinical Microbiology Reviews 25(4): 661-681.https://doi.org/10.1128/CMR.00043-12.
  16. Huttner, A., Harbarth, S., Carlet, J., Cosgrove, S., Goossens, H., Holmes, A., Jarlier, V., Voss, A., & Pittet, D. (2013). Antimicrobial resistance: a global view from the 2013 World Healthcare-Associated Infections Forum. Antimicrobial Resistance and Infection Control 1: 13. http://doi.org/10.1186/2047-2994-2-31.
  17. Keen, P.L., & Montforts, M. (2012). Antimicrobial Resistance in the Environment. Hoboken, NJ: Wiley-Blackwe.ll
  18. Khameneh, B., Diab, R., Ghazvini, K., & Bazzaz, B.S.F. (2016). Breakthroughs in bacterial resistance mechanisms and the potential ways to combat them. Microbial Pathogenesis 95: 32-42. https://doi.org/10.1016/j.micpath.2016.02.009.
  19. Khatib Haghighi, M., & Ghane, A. (2017). Study of coliform contamination of Havigh River in the west of Guilan Journal of Aquatic Caspian Sea 2(3): 55-66. (In Persian)
  20. Khatib Haghighi, S., Faeed, M., Ghane, A., & Malaki Shomali, S. (2017). Study of coliform contamination of Karganrood River in the west of Guilan province. Advanced Aquaculture Sciences Journal 1(2): 87-98. (In Persian)
  21. Kulik, N., Trapido, M., Goi, A., Veressinina, Y., & Munter, R. (2008). Combined chemical treatment of pharmaceutical effluents from medical ointment production. Chemosphere 70(8): 1525-1531. https://doi.org/10.1016/j.chemosphere.2007.08.026.
  22. Madigan, M.T., Bender, K.S., Buckley, D.H., Sattley, W.M., & Stahl, D.A. (2019). Brock Biology of Microorganisms (15th ed). Harlow Pearson Education Limited.
  23. Massey,B., Haggard, B.E., Galloway, J.M., Loftin, K.A., Meyer, M.T., & Green, W.R. (2010). Antibiotic fate and transport in three effluent-dominated Ozark streams. Ecological Engineering 36: 930–8. https://doi.org/10.1016/j.ecoleng.2010.04.009.
  24. Mirhoseini, S.H., Nikaeen, M., Khanahmad, H., & Hassanzadeh, A. (2016). Occurrence of airborne vancomycin-and gentamicin-resistant bacteria in various hospital wards in Isfahan, Iran. Advanced Biomedical Research 5: 143-148. URL: https://www.advbiores.net/text.asp?2016/5/1/143/187399.
  25. Motta, S.S., Cluzel, P., & Aldana, M. (2015). Adaptive resistance in bacteria requires epigenetic inheritance, genetic noise, and cost of efflux pumps. PloS One 10(3): p.e0118464. https://doi.org/10.1371/journal.pone.0118464.
  26. Pegler,, & Healy, B. (2007). In patients allergic to penicillin, consider second and third generation cephalosporins for life threatening infections. BMJ 335(7627): 991-991. http://doi.org/10.1136/bmj.39372.829676.47.
  27. Rezaei, P. (2018). Determination of flood catchment area using GIS and HEC-RAS hydraulic model (Case study: Goharrood River in the city of Rasht). Geography and Environmental Hazards 27: 41-56. (In Persian). http://doi.org/10.22067/geo.v0i0.69052.
  28. 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- https://doi.org/10.1016/j.scitotenv.2017.08.116.
  29. Safari Sinegani, A.A., Sharifi, Z., &Safari Sinegani, M. (2011). Experimental Methods in Soil Microbiology. (1st ed), Bu Ali Sina University Press, Hamadan, 457 p. (In Pesian)
  30. Safaeian, S., Moghadam, Z., Hosseini, H., & Ismaili, A. (2013). Antibiotic resistance in gram negative bacteria isolated from intestinal organ of Anzali Wetland wild carp. Journal of Environmental Science and Technology 15(4): 65-75. (In Persian)
  31. Schmidt, A.S., Bruun, M.S., Dalsgaard, I., Pedersen, K., & Larsen, J.L. (2000). Occurrence of antimicrobial resistance in fish-pathogenic and environmental bacteria associated with four Danish rainbow trout farms. Applied and Environmental Microbiology 66: 4908–15. https://doi.org/10.1128/AEM.66.11.4908-4915.2000.
  32. Singh, R., Singh, A.P., Kumar, S., Giri, B.S., & Kim, K.H. (2019). Antibiotic resistance in major rivers in the world: a systematic review on occurrence, emergence, and management strategies. Journal of Cleaner Production 234: 1484-1505. https://doi.org/10.1016/j.jclepro.2019.06.243.
  33. Temmerman, R., Pot, B., Huys, G., & Swings, J. (2003). Identification and antibiotic susceptibility of bacterial isolates from probiotic products. International Journal of Food Microbiology 81(1): 1-10. http://doi.org/10.1016/s0168-1605(02)00162-9.
  34. Thiele-Bruhn, S., Seibicke, T., Schulten, H.R., & Leinweber, P. (2004). Sorption of sulfonamide pharmaceutical antibiotics on whole soils and particle-size fractions. Journal of Environmental Quality 33: 1331-42. http://doi.org/10.2134/jeq2004.1331.
  35. Tripathi, K., & Sharma, A.K. (2011). Seasonal variation in bacterial contamination of water sources with antibiotic resistant faecal coliforms in relation to pollution. Journal of Applied and Natural Science 3(2): 298-302.https://doi.org/10.31018/jans.v3i2.202.
  36. Van Den Bogaard, A.E.J.M., London, N., & Stobberingh, N.N. (2000). Antimicrobial resistance in pig faecal samples from The Netherlands (five abattoirs) and Sweden. Journal of Antimicrobial Chemotherapy 45: 663-671. http://doi.org/10.1093/jac/45.5.663.
  37. Wang,, Zhou, S., Han, X., Zhang, L., Ding, S., Li, Y., Zhang, D., & Zarin, K. (2020). Occurrence, distribution, and source track of antibiotics and antibiotic resistance genes in the main rivers of Chongqing city, southwest China. Journal of Hazardous Materials 389: 122110. http://doi.org/10.1016/j.jhazmat.2020.122110.
  38. Watkinson,J., Micalizzi, G.B., Graham, G.M., Bates, J.B., & Constanzo, S.D. (2007). Antibiotic resistant Escherichia coli in wastewaters, surface waters and oysters from an urban riverine system. Applied and Environmental Microbiology 73: 5667–5670. http://doi.org/10.1128/AEM.00763-07.
  39. Zhang,, Fu, D., Xu, Y., & Liu, C. (2010). Optimization of parameters on photocatalytic degradation of chloramphenicol using TiO2 as photocatalyist by response surface methodology. Journal of Environmental Sciences 22(8): 1281-1289. http://doi.org/10.1016/s1001-0742(09)60251-5.
CAPTCHA Image