Advertisement

Annals of Microbiology

, Volume 69, Issue 9, pp 885–893 | Cite as

Source tracking of Shiga-like toxin-producing Escherichia coli in the fresh vegetable production system of South India

  • Periasamy Pushpakanth
  • Zachariah John Kennedy
  • Dananjeyan BalachandarEmail author
Original Article

Abstract

Purpose

Numerous outbreaks of foodborne diseases through fresh agricultural produce urge research to assess the source of entry of pathogens to the produce that compromise microbiological safety. In the present investigation, the entry of shiga-like toxin-producing Escherichia coli O157:H7 in a fresh vegetable production system was assessed by microbiological and molecular approaches.

Methods

Five major vegetables, viz., beetroot, cabbage, carrot, onion, parsley, and potato, being cultivated routinely in the Western Guats of South India (The Nilgiris), were assessed for the prevalence of E. coli O157:H7. The fresh produce, rhizosphere soil, and water resources were sampled and the total coliforms and E. coli counts were assessed by plate count method and the O157:H7 by polymerase chain reaction targeting shiga-like toxin gene (stx1).

Results

The results revealed that all the vegetables collected from the fields had high levels of total coliforms (3 log CFU per g) with high proportions of E. coli (1–2 log CFU per g). The prevalence of O157:H7 among the E. coli isolates in these vegetables ranged from 0 to 5.8%. However, the prevalence of O157:H7 in rhizosphere soil of these vegetables was relatively high (1.6 to 42.5%). The water used for irrigation and washing the produce (carrot) also showed the presence of O157:H7. The real-time quantitative PCR (qPCR)–based detection of stx1 revealed that the O157:H7 prevalence in these vegetables and their rhizosphere soil were in higher magnitude than the counts by culturable method. The rhizosphere soil and water samples had higher O157:H7 CFU equivalents than fresh produce.

Conclusions

It is evident that the soil as well as the irrigation and process water got contaminated with feces, which are assumed to be the primary source and cause for the entry of O157:H7 to the fresh vegetable. Hence, good agronomical practices and good hygiene post-harvest practices have to be imposed in the vegetable production system to avoid the pathogen entry.

Keywords

Fresh produce E. coli O157:H7 Irrigation water Rhizosphere stx1 gene Real-time PCR 

Notes

Funding

This work was supported by Department of Biotechnology, New Delhi through R & D Project (Molecular detection and quantification of Shiga-like toxin-producing Escherichia coli in fresh vegetables, sanction no. BT/PR10398/PFN/20/899/2013).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

N/A

Informed consent

N/A

References

  1. Abdul-Raouf UM, Beuchat LR, Ammar MS (1993) Survival and growth of Escherichia coli O157:H7 on salad vegetables. Appl Environ Microbiol 59:1999–2006PubMedPubMedCentralGoogle Scholar
  2. Alegbeleye OO, Singleton I, Sant’Ana AS (2018) Sources and contamination routes of microbial pathogens to fresh produce during field cultivation: a review. Food Microbiol 73:177–208.  https://doi.org/10.1016/j.fm.2018.01.003 CrossRefPubMedGoogle Scholar
  3. Armstrong GL, Hollingsworth J, Morris JG Jr (1996) Emerging foodborne pathogens: Escherichia coli O157: H7 as a model of entry of a new pathogen into the food supply of the developed world. Epidemiol Rev 18:29–51CrossRefPubMedGoogle Scholar
  4. Cardamone C, Aleo A, Mammina C, Oliveri G, Di Noto AMJ (2015) Assessment of the microbiological quality of fresh produce on sale in Sicily, Italy: preliminary results. J Biol Res 22:3.  https://doi.org/10.1186/s40709-015-0026-3 CrossRefGoogle Scholar
  5. CDC (2018) Food safety: food borne illness and germs. https://www.cdc.gov/foodsafety/foodborne-germs.html
  6. Cebula TA, Payne WL, Feng P (1995) Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR. J Clin Microbiol 33:248–250PubMedPubMedCentralGoogle Scholar
  7. Delannoy S, Beutin L, Fach P (2013) Towards a molecular definition of enterohemorrhagic Escherichia coli (EHEC): detection of genes located on O island 57 as markers to distinguish EHEC from closely related enteropathogenic E. coli strains. J Clin Microbiol 51:1083–1088.  https://doi.org/10.1128/jcm.02864-12 CrossRefPubMedPubMedCentralGoogle Scholar
  8. Denis N, Zhang H, Leroux A, Trudel R, Bietlot H (2016) Prevalence and trends of bacterial contamination in fresh fruits and vegetables sold at retail in Canada. Food Control 67:225–234.  https://doi.org/10.1016/j.foodcont.2016.02.047 CrossRefGoogle Scholar
  9. Fode-Vaughan KA, Maki JS, Benson JA, Collins MLP (2003) Direct PCR detection of Escherichia coli O157:H7. Lett Appl Microbiol 37:239–243.  https://doi.org/10.1046/j.1472-765X.2003.01386.x CrossRefPubMedGoogle Scholar
  10. Franz E, Semenov AV, Termorshuizen AJ, De Vos OJ, Bokhorst JG, Van Bruggen AHC (2008) Manure-amended soil characteristics affecting the survival of E. coli O157:H7 in 36 Dutch soils. Environ Microbiol 10:313–327.  https://doi.org/10.1111/j.1462-2920.2007.01453.x CrossRefPubMedGoogle Scholar
  11. Franz E et al (2014) Exploiting the explosion of information associated with whole genome sequencing to tackle Shiga toxin-producing Escherichia coli (STEC) in global food production systems. Int J Food Microbiol 187:57–72.  https://doi.org/10.1016/j.ijfoodmicro.2014.07.002 CrossRefPubMedGoogle Scholar
  12. Gil MI, Selma MV, López-Gálvez F, Allende A (2009) Fresh-cut product sanitation and wash water disinfection: problems and solutions. Int J Food Microbiol 134:37–45.  https://doi.org/10.1016/j.ijfoodmicro.2009.05.021 CrossRefPubMedGoogle Scholar
  13. Gutiérrez-Rodríguez E, Gundersen A, Sbodio A, Koike S, Suslow TV (2019) Evaluation of post-contamination survival and persistence of applied attenuated E. coli O157:H7 and naturally-contaminating E. coli O157:H7 on spinach under field conditions and following postharvest handling. Food Microbiol 77:173–184.  https://doi.org/10.1016/j.fm.2018.08.013 CrossRefPubMedGoogle Scholar
  14. Han Y, Linton RH (2004) Fate of Escherichia coli O157:H7 and Listeria monocytogenes in strawberry juice and acidified media at different pH values and temperatures. J Food Prot 67:2443–2449.  https://doi.org/10.4315/0362-028x-67.11.2443 CrossRefPubMedGoogle Scholar
  15. Hashem FA, Saleh MM (1999) Antimicrobial components of some cruciferae plants (Diplotaxis harra Forsk. and Erucaria microcarpa Boiss.). Phytother Res 13:329–332.  https://doi.org/10.1002/(SICI)1099-1573(199906)13:4<329::AID-PTR458>3.0.CO;2-U CrossRefPubMedGoogle Scholar
  16. Heredia N, García S (2018) Animals as sources of food-borne pathogens: a review. Anim Nutr 4:250–255.  https://doi.org/10.1016/j.aninu.2018.04.006 CrossRefPubMedPubMedCentralGoogle Scholar
  17. Hines E (2000) PCR-based testing: unraveling the mystery. Food Qual 7:22–28Google Scholar
  18. Ibekwe AM, Watt PM, Shouse PJ, Grieve CM (2004) Fate of Escherichia coli O157:H7 in irrigation water on soils and plants as validated by culture method and real-time PCR. Can J Microbiol 50:1007–1014.  https://doi.org/10.1139/w04-097 CrossRefPubMedGoogle Scholar
  19. Islam M, Doyle MP, Phatak SC, Millner P, Jiang X (2004) Persistence of Enterohemorrhagic Escherichia coli O157:H7 in soil and on leaf lettuce and parsley grown in fields treated with contaminated manure composts or irrigation water. J Food Prot 67:1365–1370.  https://doi.org/10.4315/0362-028X-67.7.1365 CrossRefPubMedPubMedCentralGoogle Scholar
  20. Jenkinson DS, Ladd JN (1981) Microbial biomass in soil, measurement and turn over. In: Paul EA, Ladd JN (eds) Soil biochemistry volume 5. Marcel Dekker, New York, pp 415–471Google Scholar
  21. Julien-Javaux F, Gérard C, Campagnoli M, Zuber S (2019) Strategies for the safety management of fresh produce from farm to fork. Curr Opin Food Sci.  https://doi.org/10.1016/j.cofs.2019.01.004
  22. Karmali MA (2004) Infection by shiga toxin-producing Escherichia coli. Mol Biotechnol 26:117–122.  https://doi.org/10.1385/mb:26:2:117 CrossRefPubMedGoogle Scholar
  23. Kennedy M et al (2004) Hospitalizations and deaths due to Salmonella infections, FoodNet, 1996–1999. Clin Infect Dis 38:S142–S148.  https://doi.org/10.1086/381580 CrossRefPubMedGoogle Scholar
  24. Kim J-H, Rhim S-R, Kim K-T, Paik H-D, Lee J-Y (2014) Simultaneous detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in low-fatted milk by multiplex PCR. Korean J Food Sci Anim Resour 34:717–723.  https://doi.org/10.5851/kosfa.2014.34.5.717 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Klein D, Loh T, Goulding R (1971) Rapid procedure to evaluate the dehydrogenase activity of soils low in organic matter. Soil Biol Biochem 3:385–387.  https://doi.org/10.1016/0038-0717(71)90049-6 CrossRefGoogle Scholar
  26. Liming SH, Bhagwat AA (2004) Application of a molecular beacon—real-time PCR technology to detect Salmonella species contaminating fruits and vegetables. Int J Food Microbiol 95:177–187.  https://doi.org/10.1016/j.ijfoodmicro.2004.02.013 CrossRefPubMedGoogle Scholar
  27. Mackay IM (2004) Real-time PCR in the microbiology laboratory. Clin Microbiol Infect 10:190–212.  https://doi.org/10.1111/j.1198-743X.2004.00722.x CrossRefPubMedGoogle Scholar
  28. Malorny B, Tassios PT, Rådström P, Cook N, Wagner M, Hoorfar J (2003) Standardization of diagnostic PCR for the detection of foodborne pathogens. Int J Food Microbiol 83:39–48.  https://doi.org/10.1016/S0168-1605(02)00322-7 CrossRefGoogle Scholar
  29. Mckillip JL, Drake M (2004) Real-time nucleic acid–based detection methods for pathogenic bacteria in food. J Food Prot 67:823–832.  https://doi.org/10.4315/0362-028x-67.4.823 CrossRefPubMedGoogle Scholar
  30. Muela A, García-Bringas JM, Arana I, Barcina I (2000) The effect of simulated solar radiation on Escherichia coli: the relative roles of UV-B, UV-A, and photosynthetically active radiation. Microb Ecol 39:65–71.  https://doi.org/10.1007/s002489900181 CrossRefPubMedGoogle Scholar
  31. Mukherjee A, Speh D, Diez-Gonzalez F (2007) Association of farm management practices with risk of Escherichia coli contamination in pre-harvest produce grown in Minnesota and Wisconsin. Int J Food Microbiol 120:296–302.  https://doi.org/10.1016/j.ijfoodmicro.2007.09.007 CrossRefPubMedGoogle Scholar
  32. Naganandhini S, Kennedy ZJ, Uyttendaele M, Balachandar D (2015) Persistence of pathogenic and non-pathogenic Escherichia coli strains in various tropical agricultural soils of India. PLoS One, e0130038 10.  https://doi.org/10.1371/journal.pone.0130038
  33. NHB (2017) National horticultural board: annual report. Ministry of Agriculture, IndiaGoogle Scholar
  34. NIN (2011) Dietary guidelines for indians. National Institute Of Nutrition, HyderabadGoogle Scholar
  35. O’Grady J, Ruttledge M, Sedano-Balbás S, Smith TJ, Barry T, Maher M (2009) Rapid detection of Listeria monocytogenes in food using culture enrichment combined with real-time PCR. Food Microbiol 26:4–7.  https://doi.org/10.1016/j.fm.2008.08.009 CrossRefPubMedGoogle Scholar
  36. Olaimat AN, Holley RA (2012) Factors influencing the microbial safety of fresh produce: a review. Food Microbiol 32:1–19.  https://doi.org/10.1016/j.fm.2012.04.016 CrossRefPubMedGoogle Scholar
  37. Olsen SJ, MacKinnon LC, Goulding JS, Bean NH, Slutsker L (2000) Surveillance for foodborne-disease outbreaks—United States, 1993–1997. MMWR CDC Surveill Summ 49:1–62PubMedGoogle Scholar
  38. Petrolini FVB, Lucarini R, Souza MG, Pires RH, Cunha WR, Martins CHG (2013) Evaluation of the antibacterial potential of Petroselinum crispum and Rosmarinus officinalis against bacteria that cause urinary tract infections. Braz J Microbiol 44:829–834CrossRefPubMedPubMedCentralGoogle Scholar
  39. Postollec F, Falentin H, Pavan S, Combrisson J, Sohier D (2011) Recent advances in quantitative PCR (qPCR) applications in food microbiology. Food Microbiol 28:848–861.  https://doi.org/10.1016/j.fm.2011.02.008 CrossRefPubMedGoogle Scholar
  40. Sehgal R, Kumar Y, Kumar S (2008) Prevalence and geographical distribution of Escherichia coli O157 in India: a 10-year survey. Trans R Soc Trop Med Hyg 102:380–383.  https://doi.org/10.1016/j.trstmh.2008.01.015 CrossRefPubMedGoogle Scholar
  41. Sharapov UM et al (2016) Multistate outbreak of Escherichia coli O157:H7 infections associated with consumption of fresh spinach: United States, 2006. J Food Prot 79:2024–2030.  https://doi.org/10.4315/0362-028x.Jfp-15-556 CrossRefPubMedGoogle Scholar
  42. Smith JL, Fratamico P (2005) Diarrhea-inducing Escherichia coli. In: Fratamico PM, Bhunia AK, Smith JL (eds) Foodborne pathogens: microbiology and molecular biology. Caister Academic Press, Norfolk, pp 357–382Google Scholar
  43. Söderström A et al (2008) A large Escherichia coli O157 outbreak in Sweden associated with locally produced lettuce. Foodborne Pathog Dis 5:339–349.  https://doi.org/10.1089/fpd.2007.0065 CrossRefPubMedGoogle Scholar
  44. Solomon EB, Yaron S, Matthews KR (2002) Transmission of Escherichia coli O157:H7 from contaminated manure and irrigation water to lettuce plant tissue and its subsequent internalization. Appl Environ Microbiol 68:397–400.  https://doi.org/10.1128/AEM.68.1.397-400.2002 CrossRefPubMedPubMedCentralGoogle Scholar
  45. Steele M, Mahdi A, Odumeru J (2005) Microbial assessment of irrigation water used for production of fruit and vegetables in Ontario, Canada. J Food Prot 68:1388–1392.  https://doi.org/10.4315/0362-028x-68.7.1388 CrossRefPubMedGoogle Scholar
  46. Su XL, Li Y (2005) Surface plasmon resonance and quartz crystal microbalance immunosensors for detection of Escherichia coli O157:H7. TransASAE 48:405–413.  https://doi.org/10.13031/2013.17919 CrossRefGoogle Scholar
  47. Wendel AM et al (2009) Multistate outbreak of Escherichia coli O157:H7 infection associated with consumption of packaged spinach, August–September 2006: the Wisconsin investigation. Clin Infect Dis 48:1079–1086.  https://doi.org/10.1086/597399 CrossRefPubMedGoogle Scholar
  48. Wong PYY, Kitts DD (2006) Studies on the dual antioxidant and antibacterial properties of parsley (Petroselinum crispum) and cilantro (Coriandrum sativum) extracts. Food Chem 97:505–515.  https://doi.org/10.1016/j.foodchem.2005.05.031 CrossRefGoogle Scholar
  49. XLSTAT (2010) XLSTAT. Addinsoft SARL, Paris http://www.xlstat.com Google Scholar
  50. Yilmaz A, Gun H, Ugur M, Turan N, Yilmaz H (2006) Detection and frequency of VT1, VT2 and eaeA genes in Escherichia coli O157 and O157:H7 strains isolated from cattle, cattle carcasses and abattoir environment in Istanbul. Int J Food Microbiol 106:213–217.  https://doi.org/10.1016/j.ijfoodmicro.2005.05.018 CrossRefPubMedGoogle Scholar

Copyright information

© Università degli studi di Milano 2019

Authors and Affiliations

  1. 1.Department of Agricultural MicrobiologyTamil Nadu Agricultural UniversityCoimbatoreIndia
  2. 2.Post Harvest Technology CentreTamil Nadu Agricultural UniversityCoimbatoreIndia

Personalised recommendations