Detection of Arcobacter and Helicobacter pylori Contamination in Organic Vegetables by Cultural and PCR Methods
The most demanded organic foods worldwide are those that are consumed fresh, such as fruits and vegetables. However, there is a knowledge gap about some aspects of organic food microbiological quality and safety. Organic fruits and vegetables are more exposed to pathogenic microorganisms due to surface contact with natural fertilizers such as animal manure, wastes and vermicompost used during farming. Therefore, the objective of this work was to study the contamination of organic fresh green leafy vegetables by two emergent pathogens, Arcobacter spp. and Helicobacter pylori. For this purpose, a total of 24 vegetable samples, 13 lettuce and 11 spinach were acquired from 10 different ecological supermarkets and greengroceries and analyzed by culture and PCR. Arcobacter spp. was detected in five samples (20%) by PCR, four spinach and one lettuce. One spinach sample was found to be also positive by culture. For H. pylori, the H. pylori VacA gene-specific band was detected in 12 vegetable samples (50%), 10 lettuces and two spinach. Isolation in the selective medium did not yield any positive result, possibly because of low contamination levels together with the presence of the organism in its viable but non-culturable form. Results showed significant levels of H. pylori and Arcobacter contamination in organic vegetables that are generally consumed raw, which seems to confirm that these foods can act as transmission vehicles to humans.
[1] EU Parliament, “Human health implications of organic food and organic agriculture,” Panel for the Future of Science and Technology, Dec. 2016.(www.europarl.europa.eu/RegData/etudes/STUD/2016/581922/EP RS_STU%282016%29581922_EN.pdf.)
[2] O. Alegbeleye, I. Singleton and A. S. Sant´Ana, “Sources and contamination routes of microbial pathogens to fresh produce during field cultivation: A review,” Food Microbiol., vol. 73, pp. 177-208, Feb. 2018.
[3] A. Rajwar, P. Srivastava and M. Sahgal, “Microbiology of fresh produce: route of contamination, detection methods, and remedy,” Crit Rev Food Sci Nutr., vol. 56, pp. 2383-2390, Oct. 2016.
[4] EU. “Commission notice on guidance document on addressing microbiological risks in fresh fruits and vegetables at primary production through good hygiene,” Official Journal of the European Union, 2017/C 163/01, May 2017.
[5] A. Mie, H. R. Andersen, S. Gunnarsson, J. Kahl, E. Kesse-Guyo et al., “Human health implications of organic food and organic agriculture: a comprehensive review,” Environ Health, vol. 16, p. 111, Oct. 2017.
[6] D. F. Maffei, E. Y. Batalha, M. Landgraf, D. W. Schaffner and B. D. G. M. Franco, “Microbiology of organic and conventionally grown fresh produce,” Braz J Microbiol., vol. 7, pp. 99-105, Dec. 2016.
[7] D. Corredor-García, S, García-Pinilla and C. M. Blanco-Lizarazo, “Systematic Review and Meta-analysis: Salmonella spp. prevalence in vegetables and fruits”, World J Microbiol Biotechnol., vol. 37, p. 47, Feb. 2021.
[8] S. de Oliveira, T. B.,Noronha and E. C. Tondo, “Salmonella spp. and Escherichia coli O157:H7 prevalence and levels on lettuce: A systematic review and meta-analysis”, Food Microbiol., vol 84, p. 103217, Dec. 2019.
[9] C. Willis, J. McLauchlin, H.Aird, C. Amar, C. Barker et al., “Occurrence of Listeria and Escherichia coli in frozen fruit and vegetables collected from retail and catering premises in England 2018- 2019”, Int J Food Microbiol., vol. 334, p. 108849, Dec. 2020.
[10] International Agency for Research on Cancer - World Health Organization, “Helicobacter pylori eradication as a strategy for preventing gastric cancer,” Working Group Reports, 2014. (https://www.iarc.fr/en/publications/pdfsonline/wrk/wrk8/Helicobacterpy loriEradication.pdf)
[11] World Health Organization, “WHO priority pathogens list for research and development of new antibiotics, Feb. 2017. (http://www.who.int/mediacentre/news/releases/2017/bacteria- antibiotics-needed/en/)
[12] S. Atapoor, F. S. Dehkordi and E. Rahimi, “Detection of Helicobacter pylori in various types of vegetables and salads,” Jundishapur Journal of Microbiology, vol. 7(5):e10013, May 2014.
[13] E. Yahaghi, F. Khamesipour, F. Mashayekhi, F. S. Dehkord et al., “Helicobacter pylori in vegetables and salads: genotyping and antimicrobial resistance properties,” BioMed Research International, vol. 2014, pp. 1–12, Aug. 2014.
[14] A. Lehner, T. Tasara and R. Stephan, “Relevant aspects of Arcobacter spp. as potential foodborne pathogen,” International Journal of Food Microbiology, vol. 102, pp. 127-135, July 2005.
[15] T. P. Ramees, K. Dhama, K. Karthik, R. S. Rathore, A. Kumar et al., “Arcobacter: an emerging food-borne zoonotic pathogen, its public health concerns and advances in diagnosis and control - a comprehensive review,” Vet Q., vol. 37, pp. 136-161, Dec. 2017.
[16] L. Otth, G. Solís, M. Wilson and H. Fernández, “Susceptibility of Arcobacter butzleri to heavy metals”, Braz. J. Microbiol., vol. 36, pp. 286-288, Sep. 2005.
[17] T. D. Hsu and J. Lee, “Global distribution and prevalence of Arcobacter in food and water,” Zoonoses Public Health, vol. 62, pp. 579-589, July 2015.
[18] A. González, I. F. Bayas and M. A. Ferrús, “Isolation, molecular identification and quinolone-susceptibility testing of Arcobacter spp. isolated from fresh vegetables in Spain,” Food Microbiology, vol. 65, pp. 279-283, Aug. 2017.
[19] F. J. Vesga, Y. Moreno, M. A. Ferrús, C. Campos and A. A. Trespalacios, “Detection of Helicobacter pylori in drinking water treatment plants in Bogotá, Colombia, using cultural and molecular techniques,” Int J Hygiene Environ Health., vol. 221, pp. 595-601, May 2018.
[20] H. I. Atabay and J. E. Corry, “The prevalence of campylobacters and arcobacters in broiler chickens,” Journal of Applied Microbiology, vol. 83, pp. 619-626, Nov. 1997.
[21] H. O. Nilsson, J. Blom, W. Abu-Al-Soud, A. Ljungh, L. P. Andersen and T. Wadström, “Effect of cold starvation, acid stress and nutrients on metabolic activity of Helicobacter pylori,” Appl Environ Microbiol., vol. 68, pp. 11-19, Jan. 2002.
[22] M. C. Pina-Pérez, A. González, Y. Moreno and M. A. Ferrús, “Helicobacter pylori detection in shellfish: a real-time quantitative polymerase chain reaction approach,” Foodborne Path Dis., vol. 16, pp. 137-143, Nov. 2018.
[23] L. Collado, I. Inza, J. Guarro and M. J. Figueras, “Presence of Arcobacter spp. in environmental waters correlates with high levels of fecal pollution,” Environ Microbiol., vol. 10, pp. 1635-1640, June 2008.
[24] A. González and M A. Ferrús, “Study of Arcobacter spp. contamination in fresh lettuce detected by different cultural and molecular methods,” Int J Food Microbiol., vol. 145, pp. 311-314, Jan. 2011.
[25] L. Hausdorf, M. Neumann, I. Bergmann, K. Sobiella, K. Mundt et al., “Occurrence and genetic diversity of Arcobacter spp. in a spinach- processing plant and evaluation of two Arcobacter -specific quantitative PCR assays,” Syst Appl Microbiol., vol. 36, pp. 235-243, June 2013.
[26] A. Mottola, E. Bonerba, G. Bozzo, P. Marchetti, G. V. Celano et al., “Occurrence of emerging foodborne pathogenic Arcobacter spp. isolated from pre-cut (ready-to-eat) vegetables,” Int J Food Microbiol., vol. 236, pp. 33-37, Nov. 2016.
[27] L. Moreno-Mesonero, Y. Moreno, J. L. Alonso and M. A. Ferrús, “Detection of viable Helicobacter pylori inside free-living amoebae in wastewater and drinking water samples from Eastern Spain,” Environmental Microbiology, vol. 19, pp. 4103-4112, Oct. 2017.
[28] P. Santiago, Y. Moreno and M. A. Ferrús, “Identification of viable Helicobacter pylori in drinking water supplies by cultural and molecular techniques,” Helicobacter, vol. 20, pp. 252-259, Aug. 2015.
[1] EU Parliament, “Human health implications of organic food and organic agriculture,” Panel for the Future of Science and Technology, Dec. 2016.(www.europarl.europa.eu/RegData/etudes/STUD/2016/581922/EP RS_STU%282016%29581922_EN.pdf.)
[2] O. Alegbeleye, I. Singleton and A. S. Sant´Ana, “Sources and contamination routes of microbial pathogens to fresh produce during field cultivation: A review,” Food Microbiol., vol. 73, pp. 177-208, Feb. 2018.
[3] A. Rajwar, P. Srivastava and M. Sahgal, “Microbiology of fresh produce: route of contamination, detection methods, and remedy,” Crit Rev Food Sci Nutr., vol. 56, pp. 2383-2390, Oct. 2016.
[4] EU. “Commission notice on guidance document on addressing microbiological risks in fresh fruits and vegetables at primary production through good hygiene,” Official Journal of the European Union, 2017/C 163/01, May 2017.
[5] A. Mie, H. R. Andersen, S. Gunnarsson, J. Kahl, E. Kesse-Guyo et al., “Human health implications of organic food and organic agriculture: a comprehensive review,” Environ Health, vol. 16, p. 111, Oct. 2017.
[6] D. F. Maffei, E. Y. Batalha, M. Landgraf, D. W. Schaffner and B. D. G. M. Franco, “Microbiology of organic and conventionally grown fresh produce,” Braz J Microbiol., vol. 7, pp. 99-105, Dec. 2016.
[7] D. Corredor-García, S, García-Pinilla and C. M. Blanco-Lizarazo, “Systematic Review and Meta-analysis: Salmonella spp. prevalence in vegetables and fruits”, World J Microbiol Biotechnol., vol. 37, p. 47, Feb. 2021.
[8] S. de Oliveira, T. B.,Noronha and E. C. Tondo, “Salmonella spp. and Escherichia coli O157:H7 prevalence and levels on lettuce: A systematic review and meta-analysis”, Food Microbiol., vol 84, p. 103217, Dec. 2019.
[9] C. Willis, J. McLauchlin, H.Aird, C. Amar, C. Barker et al., “Occurrence of Listeria and Escherichia coli in frozen fruit and vegetables collected from retail and catering premises in England 2018- 2019”, Int J Food Microbiol., vol. 334, p. 108849, Dec. 2020.
[10] International Agency for Research on Cancer - World Health Organization, “Helicobacter pylori eradication as a strategy for preventing gastric cancer,” Working Group Reports, 2014. (https://www.iarc.fr/en/publications/pdfsonline/wrk/wrk8/Helicobacterpy loriEradication.pdf)
[11] World Health Organization, “WHO priority pathogens list for research and development of new antibiotics, Feb. 2017. (http://www.who.int/mediacentre/news/releases/2017/bacteria- antibiotics-needed/en/)
[12] S. Atapoor, F. S. Dehkordi and E. Rahimi, “Detection of Helicobacter pylori in various types of vegetables and salads,” Jundishapur Journal of Microbiology, vol. 7(5):e10013, May 2014.
[13] E. Yahaghi, F. Khamesipour, F. Mashayekhi, F. S. Dehkord et al., “Helicobacter pylori in vegetables and salads: genotyping and antimicrobial resistance properties,” BioMed Research International, vol. 2014, pp. 1–12, Aug. 2014.
[14] A. Lehner, T. Tasara and R. Stephan, “Relevant aspects of Arcobacter spp. as potential foodborne pathogen,” International Journal of Food Microbiology, vol. 102, pp. 127-135, July 2005.
[15] T. P. Ramees, K. Dhama, K. Karthik, R. S. Rathore, A. Kumar et al., “Arcobacter: an emerging food-borne zoonotic pathogen, its public health concerns and advances in diagnosis and control - a comprehensive review,” Vet Q., vol. 37, pp. 136-161, Dec. 2017.
[16] L. Otth, G. Solís, M. Wilson and H. Fernández, “Susceptibility of Arcobacter butzleri to heavy metals”, Braz. J. Microbiol., vol. 36, pp. 286-288, Sep. 2005.
[17] T. D. Hsu and J. Lee, “Global distribution and prevalence of Arcobacter in food and water,” Zoonoses Public Health, vol. 62, pp. 579-589, July 2015.
[18] A. González, I. F. Bayas and M. A. Ferrús, “Isolation, molecular identification and quinolone-susceptibility testing of Arcobacter spp. isolated from fresh vegetables in Spain,” Food Microbiology, vol. 65, pp. 279-283, Aug. 2017.
[19] F. J. Vesga, Y. Moreno, M. A. Ferrús, C. Campos and A. A. Trespalacios, “Detection of Helicobacter pylori in drinking water treatment plants in Bogotá, Colombia, using cultural and molecular techniques,” Int J Hygiene Environ Health., vol. 221, pp. 595-601, May 2018.
[20] H. I. Atabay and J. E. Corry, “The prevalence of campylobacters and arcobacters in broiler chickens,” Journal of Applied Microbiology, vol. 83, pp. 619-626, Nov. 1997.
[21] H. O. Nilsson, J. Blom, W. Abu-Al-Soud, A. Ljungh, L. P. Andersen and T. Wadström, “Effect of cold starvation, acid stress and nutrients on metabolic activity of Helicobacter pylori,” Appl Environ Microbiol., vol. 68, pp. 11-19, Jan. 2002.
[22] M. C. Pina-Pérez, A. González, Y. Moreno and M. A. Ferrús, “Helicobacter pylori detection in shellfish: a real-time quantitative polymerase chain reaction approach,” Foodborne Path Dis., vol. 16, pp. 137-143, Nov. 2018.
[23] L. Collado, I. Inza, J. Guarro and M. J. Figueras, “Presence of Arcobacter spp. in environmental waters correlates with high levels of fecal pollution,” Environ Microbiol., vol. 10, pp. 1635-1640, June 2008.
[24] A. González and M A. Ferrús, “Study of Arcobacter spp. contamination in fresh lettuce detected by different cultural and molecular methods,” Int J Food Microbiol., vol. 145, pp. 311-314, Jan. 2011.
[25] L. Hausdorf, M. Neumann, I. Bergmann, K. Sobiella, K. Mundt et al., “Occurrence and genetic diversity of Arcobacter spp. in a spinach- processing plant and evaluation of two Arcobacter -specific quantitative PCR assays,” Syst Appl Microbiol., vol. 36, pp. 235-243, June 2013.
[26] A. Mottola, E. Bonerba, G. Bozzo, P. Marchetti, G. V. Celano et al., “Occurrence of emerging foodborne pathogenic Arcobacter spp. isolated from pre-cut (ready-to-eat) vegetables,” Int J Food Microbiol., vol. 236, pp. 33-37, Nov. 2016.
[27] L. Moreno-Mesonero, Y. Moreno, J. L. Alonso and M. A. Ferrús, “Detection of viable Helicobacter pylori inside free-living amoebae in wastewater and drinking water samples from Eastern Spain,” Environmental Microbiology, vol. 19, pp. 4103-4112, Oct. 2017.
[28] P. Santiago, Y. Moreno and M. A. Ferrús, “Identification of viable Helicobacter pylori in drinking water supplies by cultural and molecular techniques,” Helicobacter, vol. 20, pp. 252-259, Aug. 2015.
@article{"International Journal of Biological, Life and Agricultural Sciences:80824", author = "Miguel García-Ferrús and Ana González and María A. Ferrús", title = "Detection of Arcobacter and Helicobacter pylori Contamination in Organic Vegetables by Cultural and PCR Methods", abstract = "The most demanded organic foods worldwide are those that are consumed fresh, such as fruits and vegetables. However, there is a knowledge gap about some aspects of organic food microbiological quality and safety. Organic fruits and vegetables are more exposed to pathogenic microorganisms due to surface contact with natural fertilizers such as animal manure, wastes and vermicompost used during farming. Therefore, the objective of this work was to study the contamination of organic fresh green leafy vegetables by two emergent pathogens, Arcobacter spp. and Helicobacter pylori. For this purpose, a total of 24 vegetable samples, 13 lettuce and 11 spinach were acquired from 10 different ecological supermarkets and greengroceries and analyzed by culture and PCR. Arcobacter spp. was detected in five samples (20%) by PCR, four spinach and one lettuce. One spinach sample was found to be also positive by culture. For H. pylori, the H. pylori VacA gene-specific band was detected in 12 vegetable samples (50%), 10 lettuces and two spinach. Isolation in the selective medium did not yield any positive result, possibly because of low contamination levels together with the presence of the organism in its viable but non-culturable form. Results showed significant levels of H. pylori and Arcobacter contamination in organic vegetables that are generally consumed raw, which seems to confirm that these foods can act as transmission vehicles to humans.", keywords = "Arcobacter spp., Helicobacter pylori, organic vegetables, Polymerase Chain Reaction, PCR.", volume = "16", number = "5", pages = "52-4", }
