بررسی باکتری‌های کلی‌فرم مقاوم به آنتی‌بیوتیک در رودخانه زرجوب

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم خاک، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران.

2 گروه علوم دامی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران.

چکیده

آلودگی رودخانه­ها به باکتری­های مقاوم به آنتی­بیوتیک، سبب پخش این باکتری­ها در محیط و ورود آن­ها به زنجیره غذایی می­شود که تهدیدی جدی برای سلامت عمومی است. هدف این پژوهش، بررسی حضور کلی­فرم­های مقاوم به آنتی­بیوتیک در رودخانه زرجوب بود. برای بررسی نقش منابع آلودگی از آب رودخانه، در سه نقطه ورودی، وسط و خروجی شهر رشت در ماه­های مرداد و بهمن 1398 نمونه­برداری شد. شمارش کلی­فرم­ها در محیط کشت ائوزین-متیلن بلو آگار دارای µg mL-1 100 آنتی­بیوتیک و به روش پلیت کانت انجام شد. در این پژوهش، از آنتی­بیوتیک­های سفالکسین، جنتامایسین، داکسی­سایکلین و سیپروفلوکساسین استفاده شد. مقاومت ایشریشیا کولای (E. coli) به عنوان باکتری شاخص به آنتی­بیوتیک­ها بر اساس روش کربی-بائر مورد آزمایش قرار گرفت و قطر ناحیه بازدارندگی (ZOI) اندازه­گیری شد. بیشترین تعداد کلی­فرم­های مقاوم در برابر آنتی­بیوتیک سیپروفلوکساسین در نمونه مرداد ماه (CFU mL-1 Log 66/6) و کمترین تعداد آن­ها در برابر جنتامایسین و در نمونه بهمن ماه (CFU mL-1 Log 44/3) به دست آمد. همچنین بالاترین تعداد کلی­فرم­های مقاوم در برابر آنتی­بیوتیک­ها در نقطه نمونه­برداری وسط شهر و در مرداد ماه به دست آمد. روند تغییر کلی­فرم­های مقاوم به آنتی­بیوتیک­ها نشان داد که در بهمن ماه با عبور رودخانه از شهر، تعداد آن­ها افزایش یافت اما در مرداد ماه در وسط شهر بیشتر از دو جایگاه دیگر بود. بررسی ZOI باکتری E. coli نشان داد که این باکتری در هر سه جایگاه نمونه­برداری در برابر همه آنتی­بیوتیک­ها بجز داکسی­سایکلین و در هر دو ماه نمونه­برداری به سفالکسین مقاوم بود. در کل، 35 درصد کلی­فرم­های رودخانه زرجوب به سیپروفلوکساسین و باکتری شاخصE. coli جدا شده از آب رودخانه (در هر سه نقطه و دو ماه نمونه­برداری) به سفالکسین مقاوم بودند. بنابراین مصرف آب رودخانه زرجوب در پایین­دست در کشاورزی و آبزی پروروی به ویژه در فصل گرم سال مناسب نیست.

کلیدواژه‌ها


عنوان مقاله [English]

Investigation of Antibiotic Resistant Coliform Bacteria in Zarjoub River

نویسندگان [English]

  • Mohammad Bagher Farhangi 1
  • Nasrin Ghorbanzadeh 1
  • Milad Amini 1
  • Shahrokh Ghovvati 2
1 Soil Science Department, Faculty of agricultural Science, University of Guilan, Rasht, Iran Rasht, P.O. Box 41635-1314, Iran.
2 Animal Science Department, Faculty of agricultural Science, University of Guilan, Rasht, Iran.
چکیده [English]

Contamination of rivers to antibiotic-resistant bacteria causes these bacteria to spread in the environment and enter the food chain, which is a serious threat to public health. The aim of this study was to investigate the presence of antibiotic resistant coliforms in the Zarjoub River. To explore the source of contamination, water sampling was carried out in three points along the river course; entry, middle and exit of Rasht city, in August 2017 and February 2018. Plate count method was adopted to enumerate coliform bacteria number in Eosin-Methylene Blue agar medium with 100 µg mL-1 of cephalexin, gentamicin, doxycycline and ciprofloxacin antibiotics. Antibiotic resistance of Escherichia coli, as an indicator bacterium, was tested based on Kirby-Bauer method and Zone of Inhibition was measured (ZOI). The mean highest number of antibiotic-resistant coliforms was against ciprofloxacin in August samples (6.66 Log CFU mL-1) and the lowest mean number of those was against gentamicin in February samples (3.44 Log CFU mL-1). Also, the highest number of antibiotic-resistant coliforms was obtained at the sampling point in the middle of the city in August. The trend of antibiotic-resistant coliforms showed that while their number increased as the river passed through the city in February, it was more than two other sampling points in the middle of the city in August. E. coli resistance pattern, based on the ZOI, showed that this bacterium was resistant to all tested antibiotics except doxycycline in all three sampling points and also was resistant to cephalexin in both months of sampling. Overall, 35% of culturable coliforms of Zarjoub River were resistant to ciprofloxacin, and E. coli strains isolated from the river water (three sampling points and two months) were resistant to cephalexin. Therefore, downstream use of Zarjoub River water is not suitable for agriculture and aquaculture, especially in the warm season.

کلیدواژه‌ها [English]

  • Escherichia coli
  • Cephalexin
  • Ciprofloxacine
  • Guilan
  • ZOI
APHA (American Public Health Association). (2005) Standard Methods for the Examination of Water and Wastewater, (21st ed). American Water Works Association, Water Environment Federation, Washington, DC.
Awad, Y.M., Kim, S.C., Abd El-Azeem, S.A.M., Kim, K.H., Kim, K.R., Kim, K., Jeon, C., Lee, S.S., and Ok, Y.S. (2014). Veterinary antibiotics contamination in water, sediment, and soil near a swine manure composting facility. Environmental Earth Science, 71, 1433–1440.
Ayandiran, T.A., Ayandele, A.A., Dahunsi, S.O. and Ajala, O.O. (2014). Microbial assessment and prevalence of antibiotic resistance in polluted Oluwa River, Nigeria. The Egyptian Journal of Aquatic Research, 40(3), 291-299.
Calero-Caceres, W., Mendez, J., Martín-Díaz, J., and Muniesa, M. (2017). The occurrence of antibiotic resistance genes in a Mediterranean river and their persistence in the riverbed sediment. Environmental Pollution, 223, 384-394.
Chen, Z., Yu, D., He, S., Ye, H., Zhang, L., Wen, Y., Zhang, W., Shu, L. and Chen, S. (2017). Prevalence of antibiotic-resistant Escherichia coli in drinking water sources in Hangzhou City. Frontiers in Microbiology, 8, 1133.
Chopra, I., and Roberts, M. (2001). Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiology and Molecular Biology Reviews, 65, 232–260.
CLSI. (2014). Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fourth Informational Supplement. Clinical Labaraotory Standard Institute.
Dang, H., Song, L., Chen, M., and Chang, Y. (2006). Concurrence of cat and tet genes in multiple antibiotic-resistant bacteria isolated from a sea cucumber and sea urchin mariculture farm in China. Microbial Ecology, 52, 634–643.
Danner, M.C., Robertson, A., Behrends, V., and Reiss, J. (2019). Antibiotic pollution in surface fresh waters: occurrence and effects. Science of the Total Environment, 664, 793-804.
Dhawde, R., Macaden, R., Saranath, D., Nilgiriwala, K., Ghadge, A. and Birdi, T. (2018). Antibiotic resistance characterization of environmental E. coli isolated from River Mula-Mutha, Pune District, India. International Journal of Environmental Research and Public Health, 15(6), 1247.
Diwan, V., Hanna, N., Purohit, M., Chandran, S., Riggi, E., Parashar, V., Tamhankar, A.J. and 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.
Ebrahimi E., Asadi, H., Tajrishy, M., Farhangi, M.B., and Tajdari, K. (2019). Determination of cholorid and total dissolved solids in inlet rivers the Anzali lagoon. In:16th Iranian Soil Science Congress, 27-29 Aug, University of Zanjan, Iran. (In Persian)
Fajardo, A., Martinez-Martin, N., Mercadillo, M., Galan, J.C., Ghysels, B., Matthijs, S., Cornelis, P., Wiehlmann, L., Tummler, B., Baquero, F., and Martinez, J.L. (2008). The neglected intrinsic resistome of bacterial pathogens. PLos One, 3, 1619.
Freixa, A., Ejarque, E., Crognale, S., Amalfitano, S., Fazi, S., Butturini, A., and Romaní, A.M. (2016). Sediment microbial communities rely on different dissolved organic matter sources along a Mediterranean river continuum. Limnology and Oceanography, 61, 1389-1405.
Gaskins, H., Collier, C., and Anderson, D. (2002). Antibiotics as growth promotants: mode of action. Animal Biotechnology, 13(1), 29-42.
Gelband, H., Molly Miller, P., Pant, S., Gandra, S., Levinson, J., Barter, D., White, A., and Laxminarayan, R. (2015). The state of the world's antibiotics 2015. Wound Healing Southern Africa, 8(2), 30-34.
Goni-Urriza, M., Capdepuy, M., Arpin, C., Raymond, N., Caumette, P. and Quentin, C. (2000). Impact of an urban effluent on antibiotic resistance of riverine Enterobacteriaceae and Aeromonas spp. Applied and Environmental Microbiology, 66(1), 125-132.
Guo, X.P., Liu, X., Niu, Z.S., Lu, D.P., Zhao, S., Sun, X.L., Wu, J.Y., Chen, Y.R., Tou, F.Y., Hou, L. and Liu, M. (2018). Seasonal and spatial distribution of antibiotic resistance genes in the sediments along the Yangtze Estuary, China. Environmental Pollution, 242, 576-584.
Haghighatpanah, M., Mozaffari Nejad, A.S., Mojtahedi, A., Amirmozafari, N., and Zeighami, H. (2016). Detection of extended-spectrum β-lactamase (ESBL) and plasmid-borne bla CTX-M and bla TEM genes among clinical strains of Escherichia coli isolated from patients in the north of Iran. Journal of Global Antimicrobal Resistance, 7,110–113.
Kaeseberg, T., Schubert, S., Oertel, R., Zhang, J., Berendonk, T.U. and Krebs, P. (2018). Hot spots of antibiotic tolerant and resistant bacterial subpopulations in natural freshwater biofilm communities due to inevitable urban drainage system overflows. Environmental Pollution, 242, 164-170.
Keen, P.L., and Montforts, M. (2012). Antimicrobial Resistance in the environment. Hoboken, NJ: Wiley-Blackwe.ll
Khatib Haghighi, M., and Ghane, A. (2017). Study of Coliform Contamination of Havigh River in the West of Guilan Province. Journal of Aquatic Caspian Sea, 2(3), 55-66. (In Persian).
Khatib Haghighi, S., Faeed, M., Ghane, A., and 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).
Knapp, C.W., Lima, L., Olivares-Rieumont, S., Bowen, E., Werner, D., and Graham, D.W. (2012). Seasonal variations in antibiotic resistance gene transport in the Almendares River, Havana, Cuba. Frontiers in Microbiology. 3, 396.
Koo, H.J. and Woo, G.J. (2011). Distribution and transferability of tetracycline resistance determinants in Escherichia coli isolated from meat and meat products. International Journal of Food Microbiology, 145(2-3), 407-413.
Laroche, E.P.F., Fournier, M., and Pawlak, B. (2010). Transport of antibioticresistant Escherichia coli in a public rural karst water supply. Journal of Hydrology, 392(1-2), 12-21.
Machado, A., and Bordalo, A.A. (2014). Prevalence of antibiotic resistance in bacteria isolated from drinking well water available in Guinea-Bissau (West Africa). Ecotoxicology and Environmental Safety, 106, 188–194.
Madigan, M.T., Bender, K.S., Buckley, D.H., Sattley, W.M. and Stahl, D.A. (2019). Brock biology of microorganisms (15th ed). Harlow Pearson Education Limited.
Manaia, C.M. (2017). Assessing the risk of antibiotic resistance transmission from the environment to humans: non-direct proportionality between abundance and risk. Trends in Microbiology, 25(3), 173-181.
McArthur, J.V., Fletcher, D., Tuckfield, R.C., and Baker-Austin, C. (2016). Patterns of multi-antibiotic-resistant Escherichia coli from streams with no history of antimicrobial inputs. Microbial Ecology, 72(4), 840-850.
Mohanta, T. and Goel, S. (2014). Prevalence of antibiotic-resistant bacteria in three different aquatic environments over three seasons. Environmental Monitoring and Assessment, 186(8), 5089-5100.
Nikoogoftar Ranjbar, S., Pourbabaee, A.A. and Davari, K. (2016). The frequency of antibiotic resistant coliforms isolated from sewage of Qom city, Iran. Qom University of Medical Sciences Journal, 10(10), 69-77. (In Persian)
Nourani Masouleh, H., Forghani, A. and Ramezanpour, H. (2014). Quality evaluation of the incoming water to Anzali Lagoon, International Bulletin of Water Resources and Development, 1(2), 20. (In Persian)
Poonia, S., Singh, T.S. and Tsering, D.C. (2014). Antibiotic susceptibility profile of bacteria isolated from natural sources of water from rural areas of East Sikkim. Indian journal of community medicine: official publication of Indian Association of Preventive & Social Medicine, 39(3), 156.
Purohit, M., Diwan, V., Parashar, V., Tamhankar, A.J. and Lundborg, C.S. (2020). Mass bathing events in River Kshipra, Central India-influence on the water quality and the antibiotic susceptibility pattern of commensal E. coli. Plos one, 15(3), 0229664.
Rajurkar, N.S., Nongbri, B. and Patwardhan, A.M. (2003). Physico-chemical and biological investigations of river Umshyrpi at Shilling, Meghalaya. Indian Journal of Environmental Health, 45, 83-92.
Rieke, E.L., Moorman, T.B., Douglass, E.L., and 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.
Saberinia, F., Farhangi, M.B., Yaghmaeian Mahabadi, N., and Ghorbanzadeh, N. (2021). Investigation of Gowharrood River contamination to antibiotic resistant bacteria. Journal of Water and Wastewater; Ab va Fazilab, 31(7), 145-161. (In Persian).
Safari Sinegani, A.A., Sharifi, Z. and Sinegani, M.S. (2011). Experimental methods in soil microbiology. (1st ed), Bu Ali Sina University Press, Hamadan, 457 Pages. (In Pesian)
Sayyad Ghorbani Shirin, F., and Alidoost Nedamani, S. (2017). Evaluation of microbial contamination of Nawrood River to E. coli and Coliform bacteria based on global standards. Journal of Environmental Geology, 11(40), 1-10. (In Persian).
Singh, R., Singh, A.P., Kumar, S., Giri, B.S. and 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.
Stockwell, V., and Duffy, B. (2012). Use of antibiotics in plant agriculture. Review Scientific Techniqe, 31, 199–210.
Tao, R., Ying, G.G., Su, H.C., Zhou, H.W., and Sidhu, J.P. (2010). Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China. Environmental Pollution. 158, 2101–2109.
Tripathi, K. and 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.
Van den Bogaard, A., London, N., and Stobberingh, E. (2000). Antimicrobial resistance in pig faecal samples from the Netherlands (five abattoirs) and Sweden. Journal of Antimicrobial Chemotherapy, 45(5), 663-671.
Vaz-Moreira, I., Nunes, O.C., and Manaia, C.M. (2014). Bacterial diversity and antibiotic resistance in water habitats: searching the links with the human microbiome, FEMS Microbiology Reviews, 38(4), 761-778.
Walkley, A., and Black, I.A. (1934). An examination of the Degtjareff method for determining soil organic matter, and a proposed modification of the chromic acid titration method. Soil science 37(1), 29-38.
Wang, G., Zhou, S., Han, X., Zhang, L., Ding, S., Li, Y., Zhang, D., and 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.
Watkinson, A.J., Micalizzi, G.B., Graham, G.M., Bates, J.B., and 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.
Younesi N., Safari Sinegani, A.A., and Khodakaramian, G. (2017). Detection of Beta-lactamase gene in the culturable bacteria isolated from agricultural, pasture and mining soils around mines in Hamedan, Iran, Biological Journal of Microorganism, 6(21), 35-48. (In Pesian)
Zhang, X., Li, Y., Liu, B., Wang, J., Feng, C., Gao, M., and Wang, L. (2014). Prevalence of veterinary antibiotics and antibiotic-resistant Escherichia coli in the surface water of a livestock production region in northern China, Plos One, 9, 111026.