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El Emam, Mohamed M.
Languages: English
Types: Unknown
Subjects:
Listeria monocytogenes is a common food borne pathogen which is an important contaminant found in various food factory environments, because of its ability to survive in a wide range of environmental conditions, and to grow at refrigeration temperatures. L. monocytogenes has caused both occasional outbreaks and sporadic cases of food-borne illness characterised by high mortality rates. In the UK and other European countries, there has been a conspicuous rise in the number of reported cases of “Listeriosis” recently. Hence, development of efficient and rapid methods for detection of this microorganism in various foods is of great significance for the food industry; and is needed to ensure the safety of foods that are considered at high risk of contamination. Conventional bacteriological methods (e.g. ISO 11290-1/A1) for the detection and quantification of L. monocytogenes are laborious and time consuming. Therefore, development of a rapid and reliable test capable of detecting very low numbers of the organism in ready-to-eat products is required. To address this, a phage amplification assay has been developed as a rapid method for the detection of L. monocytogenes using the broad host range phage A511. Successful development of the assay required identification of a virucide that could achieve inactivation of the phage without affecting the viability of the target cell to be detected. Several different substances were evaluated as potential virucides, and among the tested materials, tea infusions were found to be the most effective virucidal agent for this experiment. The efficacy of the new assay was tested using Stilton cheese, as a representative high risk dairy product, and a method was developed to use centrifugation to concentrate bacterial cells present in samples of half-Fraser broth enrichments. The cells were detected by using the new phage amplification assay and this combination of techniques was shown to be able to detect low numbers of cells in shorter times than can be achieved using conventional culture methods. An additional molecular identification step was also developed so that the identity of the cells detected could be confirmed using a multiplex PCR which targeted conserved regions of the Listeria 16S rDNA genes. In this assay, two amplified DNA fragments were generated confirming the presence of Listeria genus (400 bp band) and also L. monocytogenes species (287 bp band). An advantage of this combined phage-PCR method its ability to detect only viable cells in food samples. The combined assay was then tested on a wide range of spiked food samples, including Camembert cheese, pasteurised milk, minced meat, turkey meat and smoked salmon. The obtained results showed that the limit of detection was as low as 20 (± 5) cfu per 25 g, and duration needed for the detection and molecular conformation of speciation was 2 days (44 h), compared to 5 days using conventional culture methods. The combined phage-PCR assay was able to achieve a sensitive and specific identification of viable L. monocytogenes present in foods within 48 h, and therefore would allow for rapid screening of food products prior to release from the factory.
  • The results below are discovered through our pilot algorithms. Let us know how we are doing!

    • CHAPTER 1..................................................................................................... 1
    • INTRODUCTION.............................................................................................. 1
    • 1.1 History of Listeria monocytogenes........................................................................ 1
    • 1.2 Infections with Listeria monocytogenes..................................................... 3
    • 1.3 Most significant outbreaks of Listeriosis by region................................... 6
    • 1.4 PErnevsiernocnemoefnLt…ist…eri…a …mo…no…cy…tog…en…es…in…th…eF…oo…d…Pr…oc…ess…in…g……..……... 8
    • 1.5 Presence of Listeria monocytogenes in Dairy Processing............................ 9
    • 1.6 Presence of Listeria monocytogenes in Fish Processing................................ 11
    • 1.7 Characteristics of Listeria monocytogenes........................................................ 2.3.7 Precipitation of Phage Lysate Using Polyethylene Glycol (PEG)..
    • 2.4 Treatment of cells or phage with Virucide.................................................
    • 2.5 Listeria monocytogenes Phage Amplification Assays...............................
    • 2.6 Isolation of Listeria monocytogenes In Cheese..........................................
    • 2.7 Conventional Plating Procedures for Detection of Listeria spp..................
    • 2.8 Gram Stain...................................................................................................
    • 2.9 Biochemical Identification Test.................................................................
    • 2.10 Molecular Identification Methods............................................................ 2.10.1 Preparation of Bacterial Chromosomol DNA Using Lysis……..... 2.10.2 Preparation of Plaque DNA……………….................................... 53 2.10.3 Preparation of Genomic DNA....................................................... 53 2.10.4 DNA Amplification by Multiplex-Polymerase Chain Reaction 54 (mPCR) 2.10.5 Preparation of Agarose Gels and Electrophoresis of DNA…...... 55
    • CHAPTER 3..................................................................................................... 57
    • OPTIMIZATION AND EVALUATION OF DIFFERENT VIRUCIDAL 57
    • 3.1 Introduction................................................................................................. 57
    • 3.2 Results......................................................................................................... 58 3.2.1 Titration and Propagation of 23074-B1 and A511........................... 59 3.2.2 Bacteriophage Concentration and Purification................................ 63 3.2.3 Evaluation of the Activity of Virucide.............................................. 64 3.2.3.1 Treatment with Acetic Acid.................................................. 65 3.2.3.2 Treatment with Virusol......................................................... 67 3.2.3.3 Treatment with Ferrous Sulphate and Hydrogen Peroxide.… 68 3.2.3.4 Treatment with Tea Infusions................................................. 70
    • 6.2 Listeria monocytogenes Identification in Cheese.....................................
    • 6.2.1 Procedures For The Isolation of Listeria monocytogenes...........
    • 6.2.2 Conventional Culture Method (ISO 11290-1)...............................
    • 7.2 Isolation of L. monocytogenes from Food Samples..................................
    • 7.3 Confirmation of Cell Identity...................................................................
    • 7.4.1.2 Detection of L. monocytogenes In Artificially Contaminated Pasteurised Milk..................................................................
    • 7.4.1.3 Detection of L. monocytogenes In Artificially Contaminated Minced Beef
    • 7.4.1.4 Detection of L. monocytogenes In Artificially Contaminated Turkey Brest
    • 7.4.1.5 Detection of L. monocytogenes In Artificially Contaminated Smoked Salmon
    • Table 1.1: Food-borne outbreaks of human listeriosis......................................
    • Table 1.2: Listeriosis outbreak connected with seafood consumption..........
    • Figure 1.1: Illustration of CAMP test...............................................................
    • Figure 1.4 : Overview of Phage Amplification Assay.......................................
    • Nakamura, H., Hatanaka, M., Ochi, K., Nagao, M., Ogasawara, J., Hase, A., Kitase, T., Haruki, K. and Nishikawa, Y. (2004) Listeria monocytogenes isolated from cold-smoked fish products in Osaka city, Japan. International Journal of Food Microbiology 94, 323-328.
    • NCFA. (1999) Listeria monocytogenes detection in foods. 136. Nordic Committee on Food Analysis. 2nd ed. http://www.nmkl.org/.
    • Niederhauser, C., Candrin, U., Hofelein, C., Jermini, M., Buhler, P.H. and Luthy, J. (1992) Use of polymerase chain reaction for detection of Listeria monocytogenes in food. Applied Environmental Microbiology 58, 1564 - 1568.
    • Nørrung, B., Andersen, J.K. and Schlund, J. (1999) Incidence and control of Listeria monocytogenes in foods in Denmark. International Journal of Food Microbiology 53, 195-203.
    • Norton, D.M., McCamey, M.A., Gall, K., Scarlett, J.M., Boor, K. J. and Wiedmann, M. (2001a) Molecular studies on the ecology of Listeria monocytogenes in the smoked fish processing industry. Applied and Environmental Microbiology 67, 198-205.
    • Norton, D., Scarlett, J., Horton, K., Sue, D., Thimothe. J., Boor, K. and Wiedmann, M. (2001b) Characterization and pathogenic potential of Listeria monocytogenes isolates from the smoked fish industry. Applied and Environmental Microbiology 67, 646-653.
    • Nwaiwu, O. (2011) The effect of growth conditions on the surface properties of Listeria monocytogenes, PhD Thesis, University of Nottingham.
    • O'Grady, J., Ruttledge, M., Sedano-Balba, S. Smith, J.T., Majella, M. (2009) Rapid detection of Listeria monocytogenes in food using culture enrichment combined with real-time PCR. Food Microbiology 26, 4-7.
    • Oladepo, D.K., Candlish, A.A.G. and Stimson, W.H. (1992) Detection of Listeria monocytogenes using polyclonal antibody. Letters in Applied Microbiology 14, 26-29.
    • Palumbo, D.J., Borucki, K.M., Mandrell, E.R. and Gorski, L. (2003) Serotyping of Listeria monocytogenes by enzyme-linked immunosorbent assay and identification of mixed-serotype cultures by colony immunoblotting. Journal of Clinical Microbiology 41, 564-571.
    • Pan,Y., Breidt, F. and Kathariou, S. (2006) Resistance of Listeria monocytogenes biofilms to sanitizing agents in a simulated food processing environment. Applied and Environmental Microbiology 72, 7711-7717.
    • Park, D.J., Drobniewski, F., Meyer, A. and Wilson, S.M. (2003) Use of a phage-based assay for phenotypic detection of Mycobacteria directly from sputum. Journal of Clinical Microbiology 41, 680-688.
    • Parish, M.E., and Higgins, P.D. (1989) Survival of Listeria monocytogenes in low pH model broth systems. Journal Food Protection 52, 144-147
    • Patel, R. and Paya, C.V (1997) Infections in solid-organ transplant recipients. Clinical Microbiology Reviews 10, 86-124.
    • Perrin, M., Bemer, .M. and Delamare, C. (2003) Fatal case of Listeria innocua bacteremia. Journal of Clinical Microbiology 41, 5308 - 5309.
    • Peterson, B.W., Sharma, P.K., van der Mei, H.C., Busscher H.J. (2012) Bacterial cell surface damage due to centrifugal compaction. Applied and Environmental Microbiology 78, 120-125.
    • Pirie, J. H. H. 1940. The genus Listerella Pirie. Science 91: 383.
    • Poltronieri, P., De Blasi, M.D. and D'Urso, O.F. (2009) Detection of Listeria monocytogenes through real-time PCR and biosensor methods. Plant Soil and Environment 55, 363-369
    • Premaratne, R. J., Lin, W.J. and Johnson, E.A. (1991) Development of an improved chemically defined minimal medium for Listeria monocytogenes. Applied Environmental Microbiology 57, 3046-3048.
    • Pritchard, T.J., Flanders, K.J. and. Donnelly, C.W (1995) Comparison of the incidence of Listeria on equipment versus environmental sites within dairy processing plants. International Journal of Food Microbiology 26, 375- 384.
    • Rakhuba, V.D. Kolomiets, I.E., Szwajcer, E., and Novik, I.G. (2010) Bacteriophage receptors, mechanisms of phage adsorption and penetration into host cell. Polish Journal of Microbiology 59, 145- 155.
    • Ramaswamy, V., Presence, V.M., Rejitha, J.S., Leashing, M.,U., Dharsana, K.S., Prasad, S.P. and Vijila, H.M. (2007) Listeria - review of epidemiology and pathogenesis. Journal of Microbiology and Immunology Infection 40, 4 - 13.
    • Rebagliati, V., Philippi, R., Rossi, M. and Troncoso, A., (2009) Prevention of food-borne listeriosis. India Journal of Pathology and Microbiology 52, 145-149.
    • Rees, C.E.D and Dodd, C.E.R (2006) Phage for rapid detection and control of bacterial pathogens in food. Advances in Applied Microbiology 59,159- 86.
    • Rees, C.E.D. and Botaris, G. (2012) "The use of phage for detection, antibiotic sensitivity testing and enumeration" in understanding Tuberculosis - Global experiences and innovative approaches to the diagnosis, Intech. P-J Cardona (ed) Chapter 14, pp 293-306.
    • Rees, J.C. and Voorhees, K.J. (2005) Simultaneous detection of two bacterial pathogens using bacteriophage amplification coupled with matrix-
    • Riedo, F.X., Pinner, R.W., de Lourdes Tosca, M., Cartter, M.L., Graves, L.M., Reeves, M.W., Weaver, R.E., Plikaytis, B.D. and Broome, C.V. (1994) A point-source foodborne listeriosis outbreak: documented incubation period and possible mild illness. Infectious Disease Journal 170, 693- 696.
    • Rittenburg, J.H. (1990) Fundamentals of immunoassay. In Development and Application of lmmunoassay for Food Anafysis. R.H. Rittenburg (Ed.). Elsevier Science Publishers Ltd., New York, NY. pp. 29-58.
    • Roberts, J.A. and Wiedmann, M. (2003) Pathogen, host and environmental factors contributing to the pathogenesis of listeriosis. Cellular and Molecular Life Sciences 60, 904-918.
    • Rocourt, J. (1986) Bacteriophages and bacteriocins of the genus Listeria. Zentralbl. Bakteriol Microbial Hyg 1, 12-28.
    • Rodríguez-Lázaro, D., Jofré, A., Aymerich, T., Garriga, M. and Pla, M. (2005) Rapid quantitative detection of Listeria monocytogenes in salmon products: evaluation of pre- real-time PCR strategies. Journal of Food Protection 68, 1467-1471.
    • Rørvik, L.M. (2000) Listeria monocytogenes in the smoked salmon industry. International Journal of Food Microbiology 62, 183-190.
    • Rørvik, L.M., Caugant, D.A. and Yndestad, M. (1995) Contamination pattern of Listeria monocytogenes and other Listeria spp. in a salmon slaughterhouse and smoked salmon processing plant. International Journal of Food Microbiology 25, 19-27.
    • Rørvik, L.M., Skjerve, E., Knudsen, B.R. and Yndestad, M., (1997) Risk factors for contamination of smoked salmon with Listeria monocytogenes during processing. International Journal of Food Microbiology 37, 215-219.
    • Rossen, L., Holmstrom, K., Olsen, E.J. and Rasmussen, F.O. (1991) A rapid polymerase chain reaction (PCR)-based assay for the identification of Listeria monocytogenes in food samples. International Journal of Food Microbiology 14, 145-151.
    • Ruban, W.S., Sharada, R. and Banday, G.T. (2001) Physical Methods of Separation and Concentration of Microbes in Food: An Aid for Rapid Detection 9, 106-111.
    • Ryser, E.T. and Marth, H.E. (1999) Listeria, listeriosis, and food safety. In: Listeriosis in animals. pp 39-57. Second Edition. (Editor: Wesely, A.) Marcel Dekker, Inc. New York, USA.
    • Sakanaka, S., Kim, M., Taniguchi, M. and Yamamoto, T. (1989) Antibacterial substances in Japanese green tea extract against Streptococcus mutans a cariogenic bacterium. Agricultural and Biological Chemistry 53, 2307- 2311.
    • Schuchat, A., Swaminathan, B. and Broome, C. V. (1991) Epidemiology of human disease. Clinical Microbiology Reviews 4, 169-183.
    • Schmelcher, M and Loessner, J.M. (2008) Bacteriophage: Powerful Tools for the Detection of Bacterial Pathogens. In: Principles of Bacterial Detection: Biosensors, Recognition Receptors and Microsystems, pp.731-754 (Editor: Zourob, M.) Springer Science + Business Media, LLC.
    • Seeliger, H.P.R. and Jones, D. (1986) The genus Listeria. (Editors: Sneath, P.H.A., Mair, N S., Sharpe, M.E. and Holt, J.G.) Bergey's manual of systemic bacteriology. pp 1235-1245. Baltimore: Williams &Wilkins.
    • Sillankorva, S., Pleteneva, E., Shaburova, O., Santos, S., Carvalho, C., Azeredo, J., and Krylov, V. (2010) Salmonella Enteritidis bacteriophage candidates for phage therapy of poultry. Journal of Applied Microbiology 108, 1175-1186.
    • Simon, M.C., Gray, D.I. and Cook, N. (1996) DNA extraction and PCR methods for the detection of Listeria monocytogenes in cold-smoked salmon. Applied and Environmental Microbiology 662, 822-824.
    • Somers, E.B. and Wong, A.C. (2004) Efficacy of two cleaning and sanitizing combinations on Listeria monocytogenes biofilms formed at low temperature on a variety of materials in the presence of ready-to-eat meat residue. Journal of Food Protection 67, 2218-2229.
    • Somer, L. and Kashi, Y. (2003) A PCR method based on 16S rRNA sequence for simultaneous detection of the genus Listeria and the species Listeria monocytogenes in food products. Journal of Food Protection 66, 1658 - 65.
    • Sorinano, J.M., Rico, H., Molto, J.C., and Manes, J. (2001) Listeria species in raw and ready-to-eat foods from restaurants. Journal of Food Protection 64, 551-553.
    • Stanley, E.C., Richard, J.M., Rebecca, J.S., Sarah M.G., Michael, R.B., McGowan, M. and Rees, C.E. (2007) Development of a new combined rapid method using phage and PCR for detection and identification of viable Mycobacterium paratuberculosis bacteria within 48 hours. Applied and Environmental Microbiology 73, 1851-1857.
    • Stessl, B., Luf, W., Wagner, M., and Schoder, D. (2009) Performance testing of six chromogenic ALOA-type media for the detection of Listeria monocytogenes. Journal of Applied Microbiology 106, 651-659.
    • Stewart, G.S.A.B., Denyer, S.P. and Jassim, S.A.A. (1995) Antiviral and antifungal composition and method. PCT patent WO 95/22254.
    • Stewart, G.S.A.B., Jassim, S.A.A., Denyer, S.P., Rees, C.E.D., Park, S. and Rostas-Mulligan, K. (1992) Methods for rapid microbial detection. PCT patent WO 92/02633.
    • Stewart, G.S.A., Jassim, S. Denyer, S.P., Newby, P., Linley, K. and Dhir, V.K. (1998) The specific and sensitive detection of bacterial pathogens within 4 h using bacteriophage amplification. Journal of Applied Microbiology 84, 777 - 783.
    • Strauch, E., Hammerl, J., and Hertwig, S. (2007) Bacteriophages: New Tools for Safer Food? Journal fur Verbraucherschutz und Lebensmittelsicherheit 2, 138-143.
    • Supanivatin, P. Kosonpisit, K. Liamkaew, R., Saeteawand, N. and Thipayarat, A. (2012) Inhibitory effects of Listeria selective enrichment media on growth characteristics of L. ivanovii. Procedia Engineering 32, 112- 118.
    • Schuchat, A., B. Swaminathan and C. V. Broome. (1991) Epidemiology of human disease. Clinical Microbiology Review 4, 169-183.
    • Swaminathan, B. and Gerner-Smidt, P. (2007) The epidemiology of human listeriosis. Microbes and Infection 9, 1236-1243.
    • Thimothe, J., Walker, J., Suvanich, V., Gall, K.L., Moody, M.W., Wiedmann, M. (2002) Detection of Listeria in crawfish processing plants and in raw, whole crawfish and processed crawfish (Procambarus spp.). Journal of Food Protection 65, 1735-1739.
    • Walker, S.J. and Stringer, F.M. (1987) Growth of Listeria monocytogenes and Aeromonas hydrophila at chill temperatures. Journal and Applied of Bacteriology 63, 20-26.
    • Wang, C. and Hong, C. (1999) A rapid PCR-based hybridization assay for the detection of Listeria monocytogenes in channel catfish. Food Microbiology 16, 29 1-297.
    • Wang, I.N., Smith, D.L., and Young, R. (2000) Holins: the protein clocks of bacteriophage infections. Annual Reviews in Microbiology 54, 799- 825.
    • Warburton, D.W., Farber, J.M., Armstrong, A., Caldeira, R., Tiwari, N.P., Babiuk, T., LaCasse, P., and Reed, S. (1991) A Canadian comparative study of modified versions of the “FDA” and “USDA” methods for the detection of Listeria monocytogenes. Journal of Food Protection 54, 669-676.
    • Yang, H., Yang, L., Wimbrow, N. A., Jiang, X. and Sun, Y. (2007) Rapid detection of Listeria monocytogenes by nanoparticle-based immunomagnetic separation and real-time PCR. International Journal of Food Microbiology 118, 132-138.
    • Young, I., Wang, I., and Roof, W.D. (2000) Phages will out: strategies of host cell lysis. Trends in Microbiology 8, 120-128.
    • Zameer, F and Gopal, S (2010) Impact of hydrogen peroxide on growth and survival of Listeria monocytogenes biofilms. E-Journal of Chemistry 7, 1008-1012.
    • Zink, R. and Loessner, J. M. (1992) Classification of virulent and temperate bacteriophages of Listeria spp. on the basis of morphology and protein analysis. Applied and Environmental Microbiology 58, 296-302.
    • Zottola, E.A. (1994) Microbial attachment and biofilm formation: a new problem for the food industry? Food Technology 48, 107-114.
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