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Research article

Food Microbiology and Biotechnology (FMB)  |  Food Biotechnology

Development of a Screening Method and Device for the Detection of Escherichia coli from Agri-Food Production Environments and Fresh Produce

Bohyun Yun 1, Hyun-Mi An 1, Won-Bo Shim 2, Won-Il Kim 1, Nguyen Bao Hung 1, Sanghyun Han 3, Hyun-Ju Kim 1, Seungdon Lee 1 and Se-Ri Kim 1*

1Microbial Safety Team, Agro-Food Safety & Crop Protection Department, National Institution of Agricultural Science, Rural Development Administration, Wanju 55365, Republic of Korea, 2Department of Agricultural Chemistry and Food Science & Technology, Gyeongsang National University, Jinju 52828, Republic of Korea, 3Division of Research Policy, Research Policy Bureau, Rural Development Administration, Jeonju 54875, Republic of Korea

Received: September 12, 2017; Accepted: October 18, 2017

J. Microbiol. Biotechnol. 2017; 27(12): 2141-2150

Published December 28, 2017 https://doi.org/10.4014/jmb.1709.09019

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

This study was conducted to develop a screening method using Colilert-18 and a device for the detection of E. coli from agri-food production environments and fresh vegetables. The specificity and sensitivity of Colilert-18 by temperature (37°C and 44°C) were evaluated with 38 E. coli and 78 non-E. coli strains. The false-positive rate was 3.8% (3/78) and 0% (0/78) at 37°C and 44°C, respectively. The detection limit of E. coli at 37°C at <1.0 log CFU/250 ml was lower than that at 44°C. The efficiency of the developed device, which comprised an incubator equipped with a UV lamp to detect E. coli in the field, was evaluated by measuring the temperature and UV lamp brightness. The difference between the set temperature and actual temperature of the developed device was about 1.0°C. When applying the developed method and device to various samples, including utensils, gloves, irrigation water, seeds, and vegetables, there were no differences in detection rates of E. coli compared with the Korean Food Code method. For sanitary disposal of culture samples after experiments, the sterilization effect of sodium dichloroisocyanurate (NaDCC) tablets was assessed for use as a substitute for an autoclave. The addition of one tablet of NaDCC per 50 ml was sufficient to kill E. coli cultured in Colilert-18. These results show that the developed protocol and device can efficiently detect E. coli from agri-food production environments and vegetables.

Keywords

E. coli, Colilert-18, NaDCC, device, on-site detection

References

  1. CDC. 2010. Preliminary FoodNet data on the incidence of infection with pathogens transmitted commonly through food - 10 states, 2009. MMWR Morb. Mortal. Wkly. Rep. 59: 418.
    Pubmed
  2. Panel EB. 2013. Scientific opinion on the risk posed by pathogens in food of non-animal origin. Part 1 (outbreak data analysis and risk ranking of food/pathogen combinations). EFSA J. 11: 3025.
    CrossRef
  3. Team EE. 2012. The European Union summary report on trends and sources of zoonoses, zoonotic agents and foodborne outbreaks in 2010. EFSA J. 10: 2597.
  4. Olaimat AN, Holley RA. 2012. Factors influencing the microbial safety of fresh produce: a review. Food Microbiol. 32: 1-19.
    Pubmed CrossRef
  5. Venkateswaran K, Murakoshi A, Satake M. 1996. Comparison of commercially available kits with standard methods for the detection of coliforms and Escherichia coli in foods. Appl. Environ. Microbiol. 62: 2236-2243.
    Pubmed PMC
  6. Gunda NSK, Naicker S, Shinde S, Kimbahune S, Shrivastava S, Mitra S. 2014. Mobile Water Kit (MWK): a smartphone compatible low-cost water monitoring system for rapid detection of total coliform and E. coli. Anal. Methods 6: 6236-6246.
    CrossRef
  7. Kim K, Myung H. 2015. Sensor node for remote monitoring of waterborne disease-causing bacteria. Sensors 15: 10569-10579.
    Pubmed PMC CrossRef
  8. Berger S. 1991. Ability of the Colilert method to recover oxidant-stressed Escherichia coli. Lett. Appl. Microbiol. 13: 247-250.
    CrossRef
  9. Covert TC, Rice EW, Johnson SA, Berman D, Johnson CH, Mason PJ. 1992. Comparing defined-substrate coliform tests for the detection of Escherichia coli in water. J. Am. Water Works Assoc. 84: 98-104.
  10. Frampton E, Restaino L. 1993. Methods for Escherichia coli identification in food, water and clinical samples based on beta-glucuronidase detection. J. Appl. Bacteriol. 74: 223-233.
    Pubmed CrossRef
  11. Kawasaki S, Nazuka E, Bari ML, Amano Y, Yoshida M, Isshiki K. 2003. Comparison of traditional culture method with DOX system for detecting coliform and Escherichia coli from vegetables. Food Sci. Technol. Res. 9: 304-308.
    CrossRef
  12. Eccles J, Searle R, Holt D, Dennis P. 2004. A comparison of methods used to enumerate Escherichia coli in conventionally treated sewage sludge. J. Appl. Microbiol. 96: 375-383.
    Pubmed CrossRef
  13. Muirhead R, Littlejohn R, Bremer P. 2004. Evaluation of the effectiveness of a commercially available defined substrate medium and enumeration system for measuring Escherichia coli numbers in faeces and soil samples. Lett. Appl. Microbiol. 39: 383-387.
    Pubmed CrossRef
  14. Jain S, Sahanoon OK, Blanton E, Schmitz A, Wannemuehler KA, Hoekstra RM, et al. 2010. Sodium dichloroisocyanurate tablets for routine treatment of household drinking water in periurban Ghana: a randomized controlled trial. Am. J. Trop. Med. Hyg. 82: 16-22.
    Pubmed PMC CrossRef
  15. Berger CN, Sodha SV, Shaw RK, Griffin PM, Pink D, Hand P, et al. 2010. Fresh fruit and vegetables as vehicles for the transmission of human pathogens. Environ. Microbiol. 12:2385-2397.
    Pubmed CrossRef
  16. Abramson A, Benami M, Weisbrod N. 2013. Adapting enzymebased microbial water quality analysis to remote areas in low-income countries. Environ. Sci. Technol. 47: 10494-10501.
    Pubmed
  17. Griffin PM, Tauxe RV. 1991. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol. Rev. 13: 60-98.
    Pubmed CrossRef
  18. Maheux AF, Huppé V, Boissinot M, Picard FJ, Bissonnette L, Bernier J-LT, et al. 2008. Analytical limits of four β-glucuronidase and β-galactosidase-based commercial culture methods used to detect Escherichia coli and total coliforms. J. Microbiol. Methods 75: 506-514.
    Pubmed CrossRef
  19. Wang J, Qiu S, Xu X, Su W, Li P, Liang B, et al. 2015. Emergence of ONPG-negative Shigella sonnei in Shanghai, China. Diagn. Microbiol. Infect. Dis. 83: 338-340.
    Pubmed CrossRef
  20. McDaniels A, Rice E, Reyes A, Johnson C, Haugland R, Stelma G. 1996. Confirmational identification of Escherichia coli, a comparison of genotypic and phenotypic assays for glutamate decarboxylase and beta-D-glucuronidase. Appl. Environ. Microbiol. 62: 3350-3354.
    Pubmed PMC
  21. ICMSF. 1996. Shigella, pp. 280-298. In: Microorganisms in Food 5:Microbiological Specifications of Food Pathogens. Ch. 6. Blackie Academic and Professional, London.
  22. Small P, Täuber MG, Hackbarth C, Sande M. 1986. Influence of body temperature on bacterial growth rates in experimental pneumococcal meningitis in rabbits. Infect. Immun. 52: 484-487.
    Pubmed PMC
  23. Palmer CJ, Tsai Y-L, Lang AL, Sangermano LR. 1993. Evaluation of Colilert-marine water for detection of total coliforms and Escherichia coli in the marine environment. Appl. Environ. Microbiol. 59: 786-790.
    Pubmed PMC
  24. Doyle M, Schoeni J. 1984. Survival and growth characteristics of Escherichia coli associated with hemorrhagic colitis. Appl. Environ. Microbiol. 48: 855-856.
    Pubmed PMC
  25. Alonso J, Soriano A, Amoros I, Ferrus M. 1998. Quantitative determination of E. coli, and fecal coliforms in water using a chromogenic medium. J. Environ. Sci. Health A Tox. Hazard. Subst. Environ. Eng. 33: 1229-1248.
    CrossRef
  26. Yeom SC. 2015. Sample analysis system using enzyme coloring method. Korea Patent 10-2015-0068755.
  27. Lee OJ. 2014. 11. 20. Eschericha coli continuous culture detection system in water sample. Korea Patent 10-2014-1465900.
  28. Julian TR, Islam MA, Pickering AJ, Roy S, Fuhrmeister ER, Ercumen A, et al. 2015. Genotypic and phenotypic characterization of Escherichia coli isolates from feces, hands, and soils in rural Bangladesh via the Colilert Quanti-Tray system. Appl. Environ. Microbiol. 81: 1735-1743.
    Pubmed PMC CrossRef
  29. Clasen T, Edmondson P. 2006. Sodium dichloroisocyanurate (NaDCC) tablets as an alternative to sodium hypochlorite for the routine treatment of drinking water at the household level. Int. J. Hyg. Environ. Health 209: 173-181.
    Pubmed CrossRef
  30. White GC. 2010. Handbook of Chlorination and Alternative Disinfectants. John Wiley & Sons, Inc., New York.
  31. Schlosser O, Robert C, Bourderioux C, Rey M, Roubin M. 2001. Bacterial removal from inexpensive portable water treatment systems for travelers. J. Travel Med. 8: 12-18.
    Pubmed CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2017; 27(12): 2141-2150

Published online December 28, 2017 https://doi.org/10.4014/jmb.1709.09019

Copyright © The Korean Society for Microbiology and Biotechnology.

Development of a Screening Method and Device for the Detection of Escherichia coli from Agri-Food Production Environments and Fresh Produce

Bohyun Yun 1, Hyun-Mi An 1, Won-Bo Shim 2, Won-Il Kim 1, Nguyen Bao Hung 1, Sanghyun Han 3, Hyun-Ju Kim 1, Seungdon Lee 1 and Se-Ri Kim 1*

1Microbial Safety Team, Agro-Food Safety & Crop Protection Department, National Institution of Agricultural Science, Rural Development Administration, Wanju 55365, Republic of Korea, 2Department of Agricultural Chemistry and Food Science & Technology, Gyeongsang National University, Jinju 52828, Republic of Korea, 3Division of Research Policy, Research Policy Bureau, Rural Development Administration, Jeonju 54875, Republic of Korea

Received: September 12, 2017; Accepted: October 18, 2017

Abstract

This study was conducted to develop a screening method using Colilert-18 and a device for
the detection of E. coli from agri-food production environments and fresh vegetables. The
specificity and sensitivity of Colilert-18 by temperature (37°C and 44°C) were evaluated with
38 E. coli and 78 non-E. coli strains. The false-positive rate was 3.8% (3/78) and 0% (0/78) at
37°C and 44°C, respectively. The detection limit of E. coli at 37°C at <1.0 log CFU/250 ml was
lower than that at 44°C. The efficiency of the developed device, which comprised an incubator
equipped with a UV lamp to detect E. coli in the field, was evaluated by measuring the
temperature and UV lamp brightness. The difference between the set temperature and actual
temperature of the developed device was about 1.0°C. When applying the developed method
and device to various samples, including utensils, gloves, irrigation water, seeds, and
vegetables, there were no differences in detection rates of E. coli compared with the Korean
Food Code method. For sanitary disposal of culture samples after experiments, the
sterilization effect of sodium dichloroisocyanurate (NaDCC) tablets was assessed for use as a
substitute for an autoclave. The addition of one tablet of NaDCC per 50 ml was sufficient to
kill E. coli cultured in Colilert-18. These results show that the developed protocol and device
can efficiently detect E. coli from agri-food production environments and vegetables.

Keywords: E. coli, Colilert-18, NaDCC, device, on-site detection

References

  1. CDC. 2010. Preliminary FoodNet data on the incidence of infection with pathogens transmitted commonly through food - 10 states, 2009. MMWR Morb. Mortal. Wkly. Rep. 59: 418.
    Pubmed
  2. Panel EB. 2013. Scientific opinion on the risk posed by pathogens in food of non-animal origin. Part 1 (outbreak data analysis and risk ranking of food/pathogen combinations). EFSA J. 11: 3025.
    CrossRef
  3. Team EE. 2012. The European Union summary report on trends and sources of zoonoses, zoonotic agents and foodborne outbreaks in 2010. EFSA J. 10: 2597.
  4. Olaimat AN, Holley RA. 2012. Factors influencing the microbial safety of fresh produce: a review. Food Microbiol. 32: 1-19.
    Pubmed CrossRef
  5. Venkateswaran K, Murakoshi A, Satake M. 1996. Comparison of commercially available kits with standard methods for the detection of coliforms and Escherichia coli in foods. Appl. Environ. Microbiol. 62: 2236-2243.
    Pubmed KoreaMed
  6. Gunda NSK, Naicker S, Shinde S, Kimbahune S, Shrivastava S, Mitra S. 2014. Mobile Water Kit (MWK): a smartphone compatible low-cost water monitoring system for rapid detection of total coliform and E. coli. Anal. Methods 6: 6236-6246.
    CrossRef
  7. Kim K, Myung H. 2015. Sensor node for remote monitoring of waterborne disease-causing bacteria. Sensors 15: 10569-10579.
    Pubmed KoreaMed CrossRef
  8. Berger S. 1991. Ability of the Colilert method to recover oxidant-stressed Escherichia coli. Lett. Appl. Microbiol. 13: 247-250.
    CrossRef
  9. Covert TC, Rice EW, Johnson SA, Berman D, Johnson CH, Mason PJ. 1992. Comparing defined-substrate coliform tests for the detection of Escherichia coli in water. J. Am. Water Works Assoc. 84: 98-104.
  10. Frampton E, Restaino L. 1993. Methods for Escherichia coli identification in food, water and clinical samples based on beta-glucuronidase detection. J. Appl. Bacteriol. 74: 223-233.
    Pubmed CrossRef
  11. Kawasaki S, Nazuka E, Bari ML, Amano Y, Yoshida M, Isshiki K. 2003. Comparison of traditional culture method with DOX system for detecting coliform and Escherichia coli from vegetables. Food Sci. Technol. Res. 9: 304-308.
    CrossRef
  12. Eccles J, Searle R, Holt D, Dennis P. 2004. A comparison of methods used to enumerate Escherichia coli in conventionally treated sewage sludge. J. Appl. Microbiol. 96: 375-383.
    Pubmed CrossRef
  13. Muirhead R, Littlejohn R, Bremer P. 2004. Evaluation of the effectiveness of a commercially available defined substrate medium and enumeration system for measuring Escherichia coli numbers in faeces and soil samples. Lett. Appl. Microbiol. 39: 383-387.
    Pubmed CrossRef
  14. Jain S, Sahanoon OK, Blanton E, Schmitz A, Wannemuehler KA, Hoekstra RM, et al. 2010. Sodium dichloroisocyanurate tablets for routine treatment of household drinking water in periurban Ghana: a randomized controlled trial. Am. J. Trop. Med. Hyg. 82: 16-22.
    Pubmed KoreaMed CrossRef
  15. Berger CN, Sodha SV, Shaw RK, Griffin PM, Pink D, Hand P, et al. 2010. Fresh fruit and vegetables as vehicles for the transmission of human pathogens. Environ. Microbiol. 12:2385-2397.
    Pubmed CrossRef
  16. Abramson A, Benami M, Weisbrod N. 2013. Adapting enzymebased microbial water quality analysis to remote areas in low-income countries. Environ. Sci. Technol. 47: 10494-10501.
    Pubmed
  17. Griffin PM, Tauxe RV. 1991. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol. Rev. 13: 60-98.
    Pubmed CrossRef
  18. Maheux AF, Huppé V, Boissinot M, Picard FJ, Bissonnette L, Bernier J-LT, et al. 2008. Analytical limits of four β-glucuronidase and β-galactosidase-based commercial culture methods used to detect Escherichia coli and total coliforms. J. Microbiol. Methods 75: 506-514.
    Pubmed CrossRef
  19. Wang J, Qiu S, Xu X, Su W, Li P, Liang B, et al. 2015. Emergence of ONPG-negative Shigella sonnei in Shanghai, China. Diagn. Microbiol. Infect. Dis. 83: 338-340.
    Pubmed CrossRef
  20. McDaniels A, Rice E, Reyes A, Johnson C, Haugland R, Stelma G. 1996. Confirmational identification of Escherichia coli, a comparison of genotypic and phenotypic assays for glutamate decarboxylase and beta-D-glucuronidase. Appl. Environ. Microbiol. 62: 3350-3354.
    Pubmed KoreaMed
  21. ICMSF. 1996. Shigella, pp. 280-298. In: Microorganisms in Food 5:Microbiological Specifications of Food Pathogens. Ch. 6. Blackie Academic and Professional, London.
  22. Small P, Täuber MG, Hackbarth C, Sande M. 1986. Influence of body temperature on bacterial growth rates in experimental pneumococcal meningitis in rabbits. Infect. Immun. 52: 484-487.
    Pubmed KoreaMed
  23. Palmer CJ, Tsai Y-L, Lang AL, Sangermano LR. 1993. Evaluation of Colilert-marine water for detection of total coliforms and Escherichia coli in the marine environment. Appl. Environ. Microbiol. 59: 786-790.
    Pubmed KoreaMed
  24. Doyle M, Schoeni J. 1984. Survival and growth characteristics of Escherichia coli associated with hemorrhagic colitis. Appl. Environ. Microbiol. 48: 855-856.
    Pubmed KoreaMed
  25. Alonso J, Soriano A, Amoros I, Ferrus M. 1998. Quantitative determination of E. coli, and fecal coliforms in water using a chromogenic medium. J. Environ. Sci. Health A Tox. Hazard. Subst. Environ. Eng. 33: 1229-1248.
    CrossRef
  26. Yeom SC. 2015. Sample analysis system using enzyme coloring method. Korea Patent 10-2015-0068755.
  27. Lee OJ. 2014. 11. 20. Eschericha coli continuous culture detection system in water sample. Korea Patent 10-2014-1465900.
  28. Julian TR, Islam MA, Pickering AJ, Roy S, Fuhrmeister ER, Ercumen A, et al. 2015. Genotypic and phenotypic characterization of Escherichia coli isolates from feces, hands, and soils in rural Bangladesh via the Colilert Quanti-Tray system. Appl. Environ. Microbiol. 81: 1735-1743.
    Pubmed KoreaMed CrossRef
  29. Clasen T, Edmondson P. 2006. Sodium dichloroisocyanurate (NaDCC) tablets as an alternative to sodium hypochlorite for the routine treatment of drinking water at the household level. Int. J. Hyg. Environ. Health 209: 173-181.
    Pubmed CrossRef
  30. White GC. 2010. Handbook of Chlorination and Alternative Disinfectants. John Wiley & Sons, Inc., New York.
  31. Schlosser O, Robert C, Bourderioux C, Rey M, Roubin M. 2001. Bacterial removal from inexpensive portable water treatment systems for travelers. J. Travel Med. 8: 12-18.
    Pubmed CrossRef