Journal of Industrial Microbiology & Biotechnology

, Volume 40, Issue 10, pp 1105–1116 | Cite as

Effectiveness of phages in the decontamination of Listeria monocytogenes adhered to clean stainless steel, stainless steel coated with fish protein, and as a biofilm

  • Geevika J. Ganegama Arachchi
  • Andrew G. Cridge
  • Beatrice M. Dias-Wanigasekera
  • Cristina D. Cruz
  • Lynn McIntyre
  • Rachel Liu
  • Steve H. Flint
  • Anthony N. Mutukumira
Environmental Microbiology

Abstract

Listeria monocytogenes is a food-borne pathogen which causes listeriosis and is difficult to eradicate from seafood processing environments; therefore, more effective control methods need to be developed. This study investigated the effectiveness of three bacteriophages (LiMN4L, LiMN4p and LiMN17), individually or as a three-phage cocktail at ≈9 log10 PFU/ml, in the lysis of three seafood-borne L. monocytogenes strains (19CO9, 19DO3 and 19EO3) adhered to a fish broth layer on stainless steel coupon (FBSSC) and clean stainless steel coupon (SSC), in 7-day biofilm, and dislodged biofilm cells at 15 ± 1 °C. Single phage treatments (LiMN4L, LiMN4p or LiMN17) decreased bacterial cells adhered to FBSSC and SSC by ≈3–4.5 log units. Phage cocktail reduced the cells on both surfaces (≈3.8–4.5 and 4.6–5.4 log10 CFU/cm2, respectively), to less than detectable levels after ≈75 min (detection limit = 0.9 log10 CFU/cm2). The phage cocktail at ≈5.8, 6.5 and 7.5 log10 PFU/cm2 eliminated Listeria contamination (≈1.5–1.7 log10 CFU/cm2) on SSC in ≈15 min. One-hour phage treatments (LiMN4p, LiMN4L and cocktail) in three consecutive applications resulted in a decrease of 7-day L. monocytogenes biofilms (≈4 log10 CFU/cm2) by ≈2–3 log units. Single phage treatments reduced dislodged biofilm cells of each L. monocytogenes strain by ≈5 log10 CFU/ml in 1 h. The three phages were effective in controlling L. monocytogenes on stainless steel either clean or soiled with fish proteins which is likely to occur in seafood processing environments. Phages were more effective on biofilm cells dislodged from the surface compared with undisturbed biofilm cells. Therefore, for short-term phage treatments of biofilm it should be considered that some disruption of the biofilm cells from the surface prior to phage application will be required.

Keywords

Listeria phages Biofilms Low count cells Stainless steel Low temperature 

Notes

Acknowledgments

The authors acknowledge New Zealand King Salmon Limited for funding the project, and Foundation for Research, Science and Technology, New Zealand for providing a scholarship (contract no. NZKX0902) to Geevika Ganegama Arachchi. The authors also thank Richard Smith, ex-company mentor at New Zealand King Salmon Limited; Ron Fyfe, Cawthron Institute; Graeme Fox at Sealord Group Limited and Karen Wittington at New Zealand King Salmon Limited for technical support.

Supplementary material

10295_2013_1313_MOESM1_ESM.docx (579 kb)
Supplementary material 1 (DOCX 578 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2013

Authors and Affiliations

  • Geevika J. Ganegama Arachchi
    • 1
  • Andrew G. Cridge
    • 5
  • Beatrice M. Dias-Wanigasekera
    • 2
  • Cristina D. Cruz
    • 3
  • Lynn McIntyre
    • 4
  • Rachel Liu
    • 1
  • Steve H. Flint
    • 1
  • Anthony N. Mutukumira
    • 1
  1. 1.Institute of Food Nutrition and Human HealthMassey UniversityAucklandNew Zealand
  2. 2.Food Standards Australia New Zealand (FSANZ)Canberra BCAustralia
  3. 3.The New Zealand Institute for Plant and Food Research LimitedAucklandNew Zealand
  4. 4.Harper Adams UniversityShropshireUK
  5. 5.Laboratory for Evolution and Development, Biochemistry DepartmentUniversity of OtagoDunedinNew Zealand

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