Environmental Science and Pollution Research

, Volume 26, Issue 2, pp 1954–1961 | Cite as

Novel approach for controlling resistant Listeria monocytogenes to antimicrobials using different disinfectants types loaded on silver nanoparticles (AgNPs)

  • Asmaa Nady MohammedEmail author
  • Sahar Abdel Aleem Abdel Aziz
Research Article


A combined use of silver nanoparticles (AgNPs) with different types of disinfectants as antimicrobial might be useful in mitigating the problem of development of bacterial resistance with a strong enhancement of the biocidal effect of disinfectants. To evaluate the biocidal activity of silver nanoparticles and its loaded forms, five commercial disinfectants (quaternary ammonium compounds (benzalkonium chloride (BC) and TH4+), Virkon®S, sodium hypochlorite, and hydrogen peroxide (H2O2)) were used against Listeria monocytogenes (L. monocytogenes) isolates at different concentrations and exposure times to reveal intra-species variability and the percentage of resistance to antimicrobial agents used. Therefore, a total of 260 specimens from animal and human stool as well as environmental samples from dairy cattle farms were cultured for isolation of L. monocytogenes. Thereafter, bacterial isolates were identified using PCR. Silver nanoparticle was synthesized using chemical reduction. Both silver nanoparticles and its loaded forms were characterized by transmission electron microscopy (TEM). The sensitivity test of 60 strains of L. monocytogenes bacteria to AgNPs and its loaded forms was evaluated using broth macrodilution method. Virkon®S/AgNPs 2.0% exhibited the highest bactericidal effect (100%) against L. monocytogenes strains followed by H2O2/AgNPs 5.0% and TH4+/AgNPs 1.0% (90% each). Furthermore, the percentage of resistance of L. monocytogenes was 0.0% to both H2O2/AgNPs 5.0% and Virkon®S/AgNPs 2.0%. In conclusion, monitoring the main source of contamination with Listeria monocytogenes in dairy cattle farms is an essential factor to achieve an efficient control. Moreover, the use of the disinfectants, Virkon®S 2.0%, H2O2 5.0%, and TH4+1.0%, loaded on silver nanoparticles composite had the strong bactericidal effect against L. monocytogenes.


Listeria monocytogenes Antimicrobial resistance AgNPs Disinfectants Disinfectants/silver nanoparticles composite 



The authors would like to thank all the members of the animal farms especially the farm workers (livestock contact) for helping us in sample collection.


This work was financially supported by Projects Funding and Granting Unit in Beni-Suef University, Beni-Suef, Egypt (the 6th stage of competitive projects, grant number 3).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

This protocol was performed by following the animal ethics guidelines and approved by both Institutional Animal Care and Use Committee (IACUC), and Institutional Review Board (IRB) of Beni-Suef University (Reference number: IORG 0009255).


  1. Abu Al-Soud W, Rådström P (1998) Capacity of nine thermostable DNA polymerases to mediate DNA amplification in the presence of PCR-inhibiting samples. Appl Environ Microbiol 64(10):3748–3753Google Scholar
  2. Bell KY, Cutter CN, Sumner SS (1997) Reduction of foodborne bicarbonate, and hydrogen peroxide spray washes. Food Microbiol 14:439–448CrossRefGoogle Scholar
  3. Belluco S, Losasso C, Patuzzi I, Rigo L, Conficoni D, Gallocchio F, Cibin V, Catellani P, Segato S, Ricci A (2016) Silver as antibacterial toward Listeria monocytogenes. Front Microbiol 7:307. CrossRefGoogle Scholar
  4. Berton V, Montesi F, Losasso C, Facco DR, Toffan A, Terregino C (2015) Study of the interaction between silver nanoparticles and Salmonella as revealed by transmission electron microscopy. J Prob Health 03:123. Google Scholar
  5. Cartwright EJ, Jackson KA, Johnson SD, Graves LM, Silk BJ, Mahon BE (2013) Listeriosis outbreaks and associated food vehicles, United States, 1998-2008. Emerg Infect Dis 19:1–9CrossRefGoogle Scholar
  6. Castro H, Jaakkonen A, Hakkinen M, Korkeala H, Lindström M (2017) Longitudinal study of the occurrence, persistence and contamination routes of Listeria monocytogenes genotypes on three Finnish dairy cattle farms. Appl Environ Microbiol 84.
  7. Cesar APR, Mesquita AJ, Prado CS, Nunes IA, Almeida Filho ES (2011) Listeria spp. E Listeria monocytogenes na produção de salsichas tipo hot dog. Ci Anim Bras 12:339–352CrossRefGoogle Scholar
  8. Conficoni D, Losasso C, Cortini E, DiCesare A, Cibin V, Giaccone V, Corno G, Ricci A (2016) Resistance to biocides in Listeria monocytogenes collected in meat-process in environments. Front Microbiol 7.
  9. Cossart P, Lebreton A (2014) A trip in the “New Microbiology” with the bacterial pathogen Listeria monocytogenes. FEBS 588:2437–2445CrossRefGoogle Scholar
  10. Dickinson JH, Kroll RG, Grant KA (1995) The direct application of the polymerase chain reaction to DNA extracted from foods. Lett Appl Microbiol 20:212–216CrossRefGoogle Scholar
  11. Dobeic M, Kenda E, Mičunovič J, Zdovc I (2011) Airborne Listeria spp in the red meat processing industry. Czech J Food Sci 29:441–447CrossRefGoogle Scholar
  12. Dos Santos CA, Seckler MM, Ingle AP, Gupta I, Galdiero S, Galdiero M, Gade A, Rai M (2014) Silver nanoparticles: therapeutical uses, toxicity, and safety issues. J Pharm Sci 103:1931–1944CrossRefGoogle Scholar
  13. Esteban JI, Oporto B, Aduriz G, Juste RA, Hurtado A (2009) Faecal shedding and strain diversity of Listeria monocytogenes in healthy ruminants and swine in Northern Spain. BMC Vet Res 5:1–10.
  14. Fouladynezhad N, Afsah Hejri L, Rukayadi Y, Nakaguchi Y, Nishibuchi M, Radu S (2013) Efficiency of four Malaysian commercial disinfectants on removing Listeria monocytogenes biofilm. Int Food Res J 20(3):1485–1490Google Scholar
  15. Fox E, O'mahony T, Clancy M, Dempsey R, O'brien M, Jordan K (2009) Listeria monocytogenes in the Irish dairy farm environment. J Food Protec 72(7):1450–1456CrossRefGoogle Scholar
  16. Franci G, Falanga A, Galdiero S, Palomba L, Rai M, Morelli G, Galdiero M (2015) Silver nanoparticles as potential antibacterial agents. Molecules 20:8856–8874CrossRefGoogle Scholar
  17. Gerba CP (2015) Quaternary ammonium biocides: efficacy in application. Appl Environ Microbiol 81:464–469CrossRefGoogle Scholar
  18. Gilbert P, Moore LE (2005) Cationic antiseptics: diversity of action under a common epithet. J Appl Microbiol 99:703–715CrossRefGoogle Scholar
  19. Girma Y, Abebe B (2018) Isolation, identification and antimicrobial susceptibility of Listeria species from raw bovine milk in Debre-Birhan Town, Ethiopia. J Zoonotic Dis Public Health 2:4 Google Scholar
  20. Gravani R (1999) Incidence and control of Listeria in food-processing facilities. In: Ryser ET, Marth EH (eds) Listeria, listeriosis and food safety, 2nd edn. Marcel Dekker, New York, pp 657–709Google Scholar
  21. Gray M, Zadoks R, Fortes E, Dogan (2004) Listeria monocytogenes isolates from foods and humans form distinct but overlapping populations. Appl Environ Microbiol 70(10): 5833–5841Google Scholar
  22. Hegstad K, Langsrud S, Lunestad BT, Scheie AA, Sunde M, Yazdankhah SP (2010) Does the wide use of quaternary ammonium compounds enhance the selection and spread of antimicrobial resistance and thus threaten our health? Microb Drug Resist 16:91–104CrossRefGoogle Scholar
  23. Hellström S (2011) Contamination routes and control of listeria monocytogenes in food production. Academic dissertation. Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine University of Helsinki FinlandGoogle Scholar
  24. Ibusquiza P, Herrera J, Cabo M (2011) Resistance to benzalkonium chloride, peracetic acid and nisin during formation of mature biofilms by Listeria monocytogenes. Food Microbiol 28(3):418–425CrossRefGoogle Scholar
  25. ISO (1998) ISO 11290-2:1998 – microbiology of food and animal feeding stuffs – horizontal method for the detection and enumeration of Listeria monocytogenes. London: BSI CorporateGoogle Scholar
  26. Jana S, Pal T (2007) Synthesis, characterization and catalytic application of silver nanoshell coated functionalized polystyrene beads. J Nanosci Nanotechnol 7:2151–2156CrossRefGoogle Scholar
  27. Lara HH, Garza-Trevino EN, Ixtepan-Turrent L, Singh DK (2011) Silver nanoparticles are broad-spectrum bactericidal and virucidal compounds. J Nanobiotechnol 9:30. CrossRefGoogle Scholar
  28. Leong D, Alvarez-Ordóñez A, Jordan K (2014) Monitoring occurrence and persistence of Listeria monocytogenes in foods and food processing environments in the Republic of Ireland. Front Microbiol 5.
  29. Li Q, Mahendra S, Lyon DY, Brunet L, Liga MV, Li D, Pedro JJ, Alvarez (2008) Antimicrobial nanomaterials for water disinfection and microbial control: potential applications and implications. Water Res 42:4591–4602CrossRefGoogle Scholar
  30. Losasso C, Belluco S, Cibin V, Zavagnin P, Mièetiæ I, Gallocchio F, Zanella M, Bregoli L, Biancotto G, Ricci A (2014) Antibacterial activity of silver nanoparticles: sensitivity of different Salmonella serovars. Front Microbiol 5:227. CrossRefGoogle Scholar
  31. Lu Z, Dai T, Huang L, Kurup DB, Tegos GP, Jahnke A, Wharton T, Hamblin MR (2010) Photodynamic therapy with a cationic functionalized fullerene rescues mice from fatal wound infections. Nanomedicine 5:1525–1533CrossRefGoogle Scholar
  32. Martín B, Perich A, Gomez D, Yanguela J, Rodriguez A, Garriga M, Aymerich T (2014) Diversity and distribution of Listeria monocytogenes in meat processing plants. Food Microbiol 44:119–127CrossRefGoogle Scholar
  33. Martínez-Suárez JV, Ortiz S, López-Alonso V (2016) Potential impact of the resistance to quaternary ammonium disinfectants on the persistence of Listeria monocytogenes in food processing environments. Front Microbiol 7:638. CrossRefGoogle Scholar
  34. Meyer B (2006) Does microbial resistance to biocides create a hazard to food hygiene? Int J Food Microbiol 112:273–279CrossRefGoogle Scholar
  35. Mijnendonckx K, Leys N, Mahillon J, Silver S, Van Houdt R (2013) Antimicrobial silver: uses, toxicity and potential for resistance. Biometals 26:609–621CrossRefGoogle Scholar
  36. Mohammed AN, Abdel Aziz SA (2017) Ecological study on Listeria monocytogenes and the extent of its resistance to different disinfectants in dairy farm for improving animal health. Asian J Anim Vet Adv 12:302–310CrossRefGoogle Scholar
  37. Mohammed HO, Atwill E, Dunbar L, Ward T, McDonough P, Gonzalez R, Stipetic K (2010) The risk of Listeria monocytogenes infection in beef cattle operations. J Appl Microbiol 108:349–356CrossRefGoogle Scholar
  38. Møretrø T, Schirmer BCT, Heir E, Fagerlund A, Hjemli P, Langsrud S (2017) Tolerance to quaternary ammonium compound disinfectants may enhance growth of Listeria monocytogenes in the food industry. Int J Food Microbiol 241:215–224CrossRefGoogle Scholar
  39. Morganti M, Scaltriti E, Cozzolino P, Bolzoni L, Casadei G, Pierantoni M, Foni E, Pongolini S (2015) Processing-dependent and clonal contamination patterns of Listeria monocytogenes in the cured ham food chain revealed by genetic analysis. Appl Environ Microbiol 82:822–831CrossRefGoogle Scholar
  40. Nassirabady N, Meghdadi H, Alami A (2015) Isolation of Listeria monocytogenes of Karun River (environmental sources rural and urban) by culture and PCR assay. Int J Enter Path 3:e21829Google Scholar
  41. Ortiz S, López-Alonso V, Rodríguez P, Martínez-Suárez JV (2016) The connection between persistent, disinfectant-resistant Listeria monocytogenes strains from two geographically separate Iberian pork processing plants: evidence from comparative genome analysis. Appl Environ Microbiol 82:308–317CrossRefGoogle Scholar
  42. Owusu-Kwarteng J, Wuni A, Akabanda F, Jespersen L (2018) Prevalence and characteristics of Listeria monocytogenes isolates in raw milk, heated milk and nunu, a spontaneously fermented milk beverage, in Ghana. Beverages 4:40. CrossRefGoogle Scholar
  43. Park HJ, Kim JY, Kim J, Lee JH, Hahn JS, Gu MB, Yoon J (2009) Silver ion-mediated reactive oxygen species generation affecting bactericidal activity. Water Res 43:1027–1032CrossRefGoogle Scholar
  44. Park J, Cha S, Cho S, Park Y (2016) Green synthesis of gold and silver nanoparticles using gallic acid: catalytic activity and conversion yield toward the 4-nitrophenol reduction reaction. J Nanopart Res 18:166. CrossRefGoogle Scholar
  45. Park MV, Neigh AM, Vermeulen JP, de la Fonteyne LJ, Verharen HW, Briedé JJ, van Loveren H, de Jong WH (2011) The effect of particle size on the cytotoxicity, inflammation, developmental toxicity and genotoxicity of silver nanoparticles. Biomaterials 32(36):9810–9817CrossRefGoogle Scholar
  46. Periasamy S, Joo HS, Duong AC, Bach TH, Tan VY, Chatterjee SS, Cheung GY, Otto M (2012) How Staphylococcus aureus biofilms develop their characteristic structure. Proc Natl Acad Sci 109:1281–1286CrossRefGoogle Scholar
  47. Rai M, Kon K, Ingle A, Duran N, Galdiero S, Galdiero M (2016) Broad-spectrum bioactivities of silver nanoparticles: the emerging trends and future prospects. Appl Microbiol Biotechnol 98:1951–1961CrossRefGoogle Scholar
  48. Sambetbayev A (2016) Efficacy of disinfectants against Listeria monocytognes. MSc Thesis Green Biotechnology and Food Security Faculty of Science and Forestry University of Eastern FinlandGoogle Scholar
  49. Sileikaite J, Puiso I, Prosycevas I, Tamulivicius S (2009) Investigation of silver nanoparticles formation kinetics during reduction of silver nitrate with sodium citrate. Mater Sci 15(1):21–27Google Scholar
  50. Szmacinski H, Lakowicz JR, Catchmark JM, Eid K, Anderson JP, Middendorf L (2008) Correlation between scattering properties of silver particle arrays and fluorescence enhancement. Appl Spectrosc 62:733–738CrossRefGoogle Scholar
  51. Tahoun ABMB, Abou Elez RMM, Abdelfatah EN, Elsohaby I, El-Gedawy AA, Elmoslemany AM (2017) Listeria monocytogenes in raw milk, milking equipment and dairy workers: molecular characterization and antimicrobial resistance patterns. J Glob Antimicrob Re 10:264–270CrossRefGoogle Scholar
  52. Tezel U, Pavlostathis SG (2015) Quaternary ammonium disinfectants: microbial adaptation, degradation and ecology. Curr Opin Biotechnol 33:296–304CrossRefGoogle Scholar
  53. Tiweri U, Walsh D, Rivas L, Jordan K, Duffy G (2014) Modelling the interaction of storage temperature, pH and water activity on the growth behaviour of Listeria monocytogenes in raw and pasteurized semi-soft rind washed milk cheese during storage following ripening. Food Control 42:248–256CrossRefGoogle Scholar
  54. Yun HS, Kim Y, Oh S, Jeon WM, Frank JF, Kim SH (2012) Susceptibility of Listeria monocytogenes biofilms and planktonic cultures to hydrogen peroxide in food processing environments. Biosci Biotechnol Biochem 76(11):2008–2013. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Asmaa Nady Mohammed
    • 1
  • Sahar Abdel Aleem Abdel Aziz
    • 1
  1. 1.Department of Hygiene, Zoonoses and Epidemiology, Faculty of Veterinary MedicineBeni-Suef UniversityBeni-SuefEgypt

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