Investigation of Radish and Parsley Vegetables Contamination to Antibiotic Resistant Bacteria - A Case Study, Guilan Province

Document Type : Research Paper


1 Soil Science Department, Faculty of Agriculture, Guilan University, Rasht, P.O. Box 41635-1314, Iran.

2 Soil Science Department, University of Guilan, Rasht, Iran


Manure application and irrigation practice with wastewaters in olericulture may introduce antibiotics and antibiotic resistant bacteria to soil which may contaminate edible vegatables. The aim of this study was to investigate the antibiotic resistant bacteria isolated from radish (Raphanus sativu) and parsley (Petroselinum crispum L.) and their cultivated soils. Three vegetable farms were selected in Pirbazar, Chaboksar and Fouman areas in Guilan province. Samples were taken from vegetables and soils at three replications. After making serial dilution, their heterotrophic and coliform bacteria number were determined on NA and EMB agar media respectively, containing 100 µg/mL antibiotic. Four antibiotics including cephalexin, ciprofloxacin, gentamicin and trimethoprim were tested. Antibiotic resistant index (ARI) was calculated by dividing bacteria colony numbers on each antibiotic contained medium to control (medium without antibiotic). Data were analyzed as split-split plot design with three locations as main plot sector, two olericulture fields as sub-plot and four antibiotics as sub-sub plot factor. The effect of antibiotic was significant on the vegetable and soil bacteria ARI (p<0.05). But the effect of sampling area and farm (radish and parsley farms) were significant only on the heterotrophic and coliform bacteria ARI, respectively (p <0.01). In Chaboksar area, 29% of vegetable's heterotrophic bacteria and 42% of soil coliforms were resistant to gentamicin. The highest coliform ARI (38%) was obtained in radish and was to gentamicin. Overall, culturable bacteria from vegetables of Chaboksar area had higher antibiotic resistance index and eating fresh vegetables cultivated in this area is not recommended.


Alef, K. and Nannipieri, P. (1995). Methods in Applied Soil Microbiological and Biochemistry. Academic Press INC.
Aminshahidi, M., Fani, F., Firoozian, N. and Rafaatpour, N. (2018). An investigation of the contamination of vegetables and the irrigating water with bacteria causing food-borne and water-borne diseases in different areas of the shiraz in summer 95. Journal of Fasa University of Medical Sciences, 8(1), 708-717. (In Farsi)
Azanu, D., Mortey, C., Darko, G., Weisser, J. J., Styrishave, B. and Abaidoo, R. C. (2016). Uptake of antibiotics from irrigation water by plants. Chemosphere, 157, 107-114.
Baguer, A. J., Jensen, J. and Krogh, P. H. (2000). Effects of antibiotics oxytetracycline and tylosin on soil fauna. Chemosphere, 40, 751–757.
Boxall, A. B., Johnson, P., Smith, E. J., Sinclair, C. J., Stutt, E. and Levy, L.S. (2006). Uptake of veterinary medicines from soils into plants. Journal of Agricultural and Food Chemistry, 54(6), 2288-2297.
Clementi, F. and Aquilanti, L. (2011). Recent investigations and updated criteria for the assessment of antibiotic resistance in food lactic acid bacteria. Anaerobe, 17(6), 394-398.
Dingman, W. D. (2000). Growth of Escherichia coli O157:H7 in bruised apple (Malus domestica) tissue as influenced by cultivar, date of harvest, and source. Applied and Environmental Microbiology, 66, 1077-1083.
Ebrahimi, A.A., Ebrahimi, S., and Agholi, M. (2009). Evaluation of resistance of Shigellae isolated from children with diarrhea in Fasa in summer 2005. Tebbe Jonub Journal, 12 (3); 225-230. (In Farsi).
Rasouli, A., Faghihi, S.M. (2014). A survey on antibacterial drug usage in dairy farms in Tehran province during 2004-2005. Journal of Veterinary Research, 69,1:41-48. (In Farsi).
Fan, H., Brunham, R. C. and McClarty, G. (1992). Acquisition and synthesis of folates by obligate intracellular bacteria of the genus Chlamydia. Journal of Clinical Investigations, 90, 1803–11.
Fisher, T.L. and Golden D.A. (1998). Fate of Escherichia coli O157:H7 in ground apples used in cider production. Journal of Food Protection, 61,1372-1374.
Hamilton-Miller, J. M. (1988). Reversal of activity of trimethoprim against Gram-positive cocci by thymidine, thymine and ‘folates’. Journal of Antimicrobial Chemotherapy, 22, 35-9.
Hassan, S. A., Altalhi, A. D., Gherbawy, Y. A. and El-Deeb, B.A. (2011). Bacterial load of fresh vegetables and their resistance to the currently used antibiotics in Saudi Arabia. Foodborne Pathogens and Disease, 8(9), 1011-1018.
Hu, X., Zhou, Q. and Luo, Y. (2010). Occurrence and source analysis of typical veterinary antibiotics in manure, soil, vegetables and groundwater from organic vegetable bases. Northern China Environment Pollution, 158 (9), 2992–2998.
Hughes, P. and Heritage, J. (2004). Antibiotic growth-promoters in food animals. In: S. Jutzi, Assessing Quality and Safety of Animal Feeds, FAO Animal Production and Health Paper, pp.129-152. Food and Agriculture Organization of the United Nations, Rome.
Jones, D. L. (1999). Potential health risks associated with the persistence of Escherichia coli O157:H7 in agricultural environments. Soil Use and Management, 15, 76-83.
Kabir, A., Das, A. K. and Kabir, M.S. (2014). Incidence of antibiotic resistant pathogenic bacteria in vegetable items sold by local and super shops in Dhaka city. Stamford Journal of Microbiology, 4(1), 13-18.
Kumar, K., Gupta, S. C., Baidoo, S. K., Chander, Y. and Rosen, C. J. (2005). Antibiotic uptake by plants from soil fertilized with animal manure. Journal of Environmental Quality, 34, 2082–2085.
Li, D., Yang, M., Hu, J., Ren, L., Zhang, Y. and Li, K. (2008). Determination and fate of oxytetracycline and related compounds in oxytetracycline production wastewater and the receiving river. Environmental Toxicology and Chemistry, 27, 80–86.
Madigan, M.T., Clark, D.P., Stahl, D., and Martinko, J.M. (2010). Brock Biology of Microorganisms. 13th edition. Benjamin Cummings.
Martinez, J.L. (2009). Environmental pollution by antibiotics and by antibiotic resistance determinants. Environmental Pollution, 157(11), 2893-2902.
Michael, J., and Weinstein, M. J. (2011). Handbook of Antibiotic Discovery and Development (3 ed.). New York: Springer.
Ministry of Agriculture, (2009) Agricultural and Animal Production Statistics, Deputy of Economic Planning. Office of Statistics and Information Technology. (In Farsi).
Mohammed Hamzah, A., Mohammed Hussein, A., and Mahmoud Khalef, J. (2013). Isolation of Escherichia coli 0157: H7 strain from fecal samples of zoo animal. The Scientific World Journal, 2013.
Nipa, M. N., Mazumdar, R. M., Hasan, M. M., Fakruddin, M. D., Islam, S., Bhuiyan, H. R. and Iqbal, A. (2011). Prevalence of multi drug resistant bacteria on raw salad vegetables sold in major markets of Chittagong city, Bangladesh. Middle-East Journal of Scientific Research, 10(1), 70-77.
Österblad, M., Pensala, O., Peterzéns, M., Heleniusc, H. and Huovinen, P. (1999). Antimicrobial susceptibility of Enterobacteriaceae isolated from vegetables. Journal of Antimicrobial Chemotherapy, 43(4): 503-509.
Pan, M. and Chu, L. M. (2016). Phytotoxicity of veterinary antibiotics to seed germination and root elongation of crops. Ecotoxicology and Environmental Safety, 126, 228-237.
Pell, A. N. (1997). Manure and microbes: public and animal health problem. Journal of Dairy Science, 80, 2673-2681.
Phillips, I., Casewell, M., Cox, T., Groot, B. D., Friis, C., Jones, R., Nightingale, C., Preston, R. and Waddell, J. (2004). Does the use of antibiotics in food animals pose a risk to human health? A critical review of published data. Journal of Antimicrobial Chemotherapy, 53, 28-52.
Rezai, H. (2013). A review of research on application of livestock manure in agricultural land of Iran. Journal of Land Management, 1, 55-68. (In Farsi)
Rizzo, L., Manaia, C., Merlin, C., Schwartz, T., Dagot, C., Ploy, M. C., Michael, I. and 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.
Safari Sinegani, A. A., Hasanalizadeh, N. and Rahimi, Gh. (2018). The effect of irrigation with unrefined municipal wastewater on the fecal coliforms number in soil and vegetables. Water and Soil Science, 28(4), 57-68. (In Farsi)
Schwaiger, K., Helmke, K., Hölzel, C. S. and Bauer, J. (2011). Comparative analysis of the bacterial flora of vegetables collected directly from farms and from supermarkets in Germany. International Journal of Environmental Health Research, 21(3), 161–172.
Soltan Dallal, M. M., Nikmanesh, B., Haghi-Ashtiani, M. T., Okazi, A. and Sharifi Yazdi, M. K. (2017). Serotyping and multiple antibiotic resistance patterns of Shigella sonnei isolated from diarrhea in Children’s Medical Center of Tehran. Journal of Paramedical Faculty of Tehran University of Medical Sciences (Payavard Salamat), 11(5), 560-566. (In Farsi)
Stewart, P. S. and Costerton, J. W. (2001), Antibiotic resistance of bacteria in biofilms. The Lancet, 358(9276): pp.135-138.
Tasho, R. P. and Cho, J. Y. (2016). Veterinary antibiotics in animal waste, its distribution in soil and uptake by plants: a review. Science of the Total Environment, 563, 366-376.
Environment Agency (Standing Committee of Analysts) (2002). The Microbiology of Drinking Water– Part 1 - Water Quality and Public Health; Nottingham, Environment Agency.
US Food and Drug Administration (2011). Antimicrobial resistant bacteria in animals and food, last updated 2011.
Van Den Bogaard A. E. J. M., London N. and Stobberingh E. E. (2000). Antimicrobial resistance in pig fecal samples from The Netherlands (five abattoirs) and Sweden. Journal of Antimicrobial Chemotherapy, 45(5), 663–671.