Bacteria on Meat Abattoir Process Surfaces after Sanitation: Characterisation of Survival Properties of Listeria monocytogenes and the Commensal Bacterial Flora

Abstract

Contamination of food with spoilage bacteria and pathogens from food processing environment remains a challenge for the food industry. Bacteria able to persist in such environments over time must survive several hygienic hurdles. The aim of this study was to identify bacteria surviving practical disinfection and compare their survival abilities with representative isolates of the pathogen Listeria monocytogenes. Bacteria isolated from processing surfaces after cleaning and disinfection in a meat abattoir were identified. Selected isolates of the most frequently isolated bacterial genera along with eight meat associated L. monocytogenes were further characterized with regard to biofilm formation abilities at 12 and 20, tolerance to desiccation (stainless steel at 70% RH at 12) and bactericidal effects of recommended in-use-concentrations of four commercial disinfectants on stainless steel surface. The most dominating bacterial genera based on counts on non-selective agar were Aerococcus, Acinetobacter, Pseudomonas, Serratia and Staphylococcus. Isolates of Citrobacter. Enterobacter and Serratia dominated on agar plates selective for Enterobacteriaceae. In general, Gram negative bacteria formed more biofilm than Gram positives, especially at 12 with the best biofilm formers being Acinetobacter, Citrobacter and Pseudomonas. Listeria monocytogenes were poor biofilm formers. Gram positives survived better air drying than Gram negatives. Strains of L. monocytogenes were more sensitive to desiccation than the other Gram positives; Aerococcus, Kocuria and Staphylococcus. Two disinfectants containing peracetic acid and a disinfectant containing alkylaminoacetate had limited or no antibacterial effect against bacteria dried on stainless steel. A quaternary ammonium compound-based disinfectant provided >2 log reductions of Aerococcus, Acinetobacter and Listeria. Only 0.5 log reductions were obtained against Staphylococcus and no bactericidal effect against Serratia. In this study the dominating flora in a meat abattoir was isolated and identified. Several of these bacteria were better biofilm formers and more resistant to desiccation and disinfection than L. monocytogenes. The disinfectants tested had limited bactericidal activity against surface associated bacteria.


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T. Møretrø, S. Langsrud and E. Heir, "Bacteria on Meat Abattoir Process Surfaces after Sanitation: Characterisation of Survival Properties of Listeria monocytogenes and the Commensal Bacterial Flora," Advances in Microbiology, Vol. 3 No. 3, 2013, pp. 255-264. doi: 10.4236/aim.2013.33037.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] J. T. Holah, J. Bird and K. E. Hall, “The Microbial Ecology of High-Risk, Chilled Food Factories; Evidence for Persistent Listeria spp. and Escherichia coli Strains,” Journal of Applied Microbiology, Vol. 97, No. 1, 2004, pp. 68-77. doi:10.1111/j.1365-2672.2004.02272.x
[2] R. B. Tompkin, “Control of Listeria monocytogenes in the Food-Processing Environment,” Journal of Food Protection, Vol. 65, No. 4, 2002, pp. 709-725.
[3] T. Moretro and S. Langsrud, “Listeria monocytogenes: Biofilm Formation and Persistence in Food Processing Environments,” Biofilms, Vol. 1, No. 2, 2004, pp. 107-121. doi:10.1017/S1479050504001322
[4] M. K. Borucki, et al., “Variation in Biofilm Formation among Strains of Listeria monocytogenes,” Applied and Environmental Microbiology, Vol. 69, No. 12, 2003, pp. 7336-7342. doi:10.1128/AEM.69.12.7336-7342.2003
[5] B. Aase, et al., “Occurrence of and a Possible Mechanism for Resistance to a Quaternary Ammonium Compound in Listeria monocytogenes,” International Journal of Food Microbiology, Vol. 62, No. 1-2, 2000, pp. 57-63. doi:10.1016/S0168-1605(00)00357-3
[6] J. M. Lunden, et al., “Persistent Listeria monocytogenes Strains Show Enhanced Adherence to Food Contact Surface after Short Contact Times,” Journal of Food Protection, Vol. 63, No. 9, 2000, pp. 1204-1207.
[7] E. M. Fox, N. Leonard and K. Jordan, “Physiological and Transcriptional Characterization of Persistent and Nonpersistent Listeria monocytogenes Isolates,” Applied and Environmental Microbiology, Vol. 77, No. 18, 2011, pp. 6559-6569. doi:10.1128/AEM.05529-11
[8] J. Lundén, R. Tolvanen and H. Korkeala, “Acid and Heat Tolerance of Persistent and Nonpersistent Listeria monocytogenes Food Plant Strains,” Letters in Applied Microbiology, Vol. 46, No. 2, 2008, pp. 276-280. doi:10.1111/j.1472-765X.2007.02305.x
[9] G. Brightwell, et al., “Identifying the Bacterial Community on the Surface of Intralox(TM) Belting in a Meat Boning Room by Culture-Dependent and Culture-Independent 16S rDNA Sequence Analysis,” International Journal of Food Microbiology, Vol. 109, No. 1-2, 2006, pp. 47-53. doi:10.1016/j.ijfoodmicro.2006.01.008
[10] E. Mettler and B. Carpentier, “Variations over Time of Microbial Load and Physicochemical Properties of Floor Materials after Cleaning in Food Industry Premises,” Journal of Food Protection, Vol. 61, No. 1, 1998, pp. 57-65.
[11] M. Sharma and S. K. Anand, “Characterization of Constitutive Microflora of Biofilms in Dairy Processing Lines,” Food Microbiology, Vol. 19, No. 6, 2002, pp. 627-636. doi:10.1006/fmic.2002.0472
[12] D. Bagge-Ravn, et al., “The Microbial Ecology of Processing Equipment in Different Fish Industries—Analysis of the Microflora during Processing and Following Cleaning and Disinfection,” International Journal of Food Microbiology, Vol. 87, No. 3, 2003, pp. 239-250. doi:10.1016/S0168-1605(03)00067-9
[13] B. Guobjornsdottir, H. Einarsson and G. Thorkelsson, “Microbial Adhesion to Processing Lines for Fish Fillets and Cooked Shrimp: Influence of Stainless Steel Surface Finish and Presence of Gram-Negative Bacteria on the Attachment of Listeria monocytogenes,” Food Technology and Biotechnology, Vol. 43, No. 1, 2005, pp. 55-61.
[14] S. Langsrud, L. Seifert and T. Moretro, “Characterization of the Microbial Flora in Disinfecting Footbaths with Hypochlorite,” Journal of Food Protection, Vol. 69, No. 9, 2006, pp. 2193-2198.
[15] T. Moretro, et al., “Evaluation of the Antimicrobial Effect of a Triclosan-Containing Industrial Floor Used in the Food Industry,” Journal of Food Protection, Vol. 69, No. 3, 2006, pp. 627-633.
[16] D. K. Jeong and J. F. Frank, “Growth of Listeria monocytogenes at 10℃ in Biofilms with Microorganisms Isolated from Meat and Dairy Processing Environments,” Journal of Food Protection, Vol. 57, No. 7, 1994, pp. 576-586.
[17] V. Leriche and B. Carpentier, “Limitation of Adhesion and Growth of Listeria monocytogenes on Stainless Steel Surfaces by Staphylococcus sciuri Biofilms,” Journal of Applied Microbiology, Vol. 88, No. 4, 2000, pp. 594-605. doi:10.1046/j.1365-2672.2000.01000.x
[18] B. Carpentier and D. Chassaing, “Interactions in Biofilms between Listeria monocytogenes and Residant Microorganisms from Food Industry Premises,” International Journal of Food Microbiology, Vol. 97, No. 2, 2004, pp. 111-122. doi:10.1016/j.ijfoodmicro.2004.03.031
[19] O. Habimana, et al., “Enhanced Surface Colonization by Escherichia coli O157:H7 in Biofilms Formed by an Acinetobacter calcoaceticus Isolate from Meat-Processing Environments,” Applied and Environmental Microbiology, Vol. 76, No. 13, 2010, pp. 4557-4559. doi:10.1128/AEM.02707-09
[20] B. C. Schirmer, E. Heir and S. Langsrud, “Characterization of the Bacterial Spoilage Flora in Marinated Pork Products,” Journal of Applied Microbiology, Vol. 106, No. 6, 2009, pp. 2106-2116. doi:10.1111/j.1365-2672.2009.04183.x
[21] E. Fugett, et al., “International Life Sciences Institute North America Listeria monocytogenes Strain Collection: Development of Standard Listeria monocytogenes Strain Sets for Research and Validation Studies,” Journal of Food Protection, Vol. 69, No. 12, 2006, pp. 2929-2938.
[22] P. Glaser, et al., “Comparative Genomics of Listeria Species,” Science, Vol. 294, No. 5543, 2001, pp. 849-852.
[23] T. Moretro, et al., “Factors Affecting Survival of Shigatoxin-Producing Escherichia coli on Abiotic Surfaces,” International Journal of Food Microbiology, Vol. 138, No. 1-2, 2010, pp. 71-77.
[24] European Committee for standardization, “NS-EN 13697: 2001: Quantitative Non-Porous Surface Test for the Evaluation of Bactericidal and/or Fungicidal Activity of Chemical Disinfectants Used in Food, Industrial, Domestic and Institutional Areas. Test Method and Requirements without Mechanical Action,” 2001.
[25] T. Moretro, et al., “Evaluation of Efficacy of Disinfectants against Salmonella from the Feed Industry,” Journal of Applied Microbiology, Vol. 106, No. 3, 2009, pp. 1005-1012. doi:10.1111/j.1365-2672.2008.04067.x
[26] B. Carpentier and O. Cerf, “Review—Persistence of Listeria monocytogenes in Food Industry Equipment and Premises,” International Journal of Food Microbiology, Vol. 145, No. 1, 2011, pp. 1-8. doi:10.1016/j.ijfoodmicro.2011.01.005
[27] K. Sasahara and E. Zottola, “Biofilm Formation by Listeria monocytogenes Utilizes a Primary Colonizing Microorganism in Flowing Systems,” Journal of Food Protection, Vol. 56, No. 12, 1993, pp. 1022-1028.
[28] D. E. Norwood and A. Gilmour, “The Growth and Resistance to Sodium Hypochlorite of Listeria monocytogenes in a Steady-State Multispecies Biofilm,” Journal of Applied Microbiology, Vol. 88, No. 3, 2000, pp. 512-520. doi:10.1046/j.1365-2672.2000.00990.x
[29] T. Combrouse, et al., “Quantification of the Extracellular Matrix of the Listeria monocytogenes Biofilms of Different Phylogenic Lineages with Optimization of Culture Conditions,” Journal of Applied Microbiology, Vol. 114, No. 4, 2013, pp. 1120-1131. doi:10.1111/jam.12127
[30] A. Moltz and S. Martin, “Formation of Biofilms by Listeria monocytogenes under Various Growth Conditions,” Journal of Food Protection, Vol. 68, No. 1, 2005, pp. 92-97.
[31] J. Folsom, G. Siragusa and J. Frank, “Formation of Biofilm at Different Nutrient Levels by Various Genotypes of Listeria monocytogenes,” Journal of Food Protection, Vol. 69, No. 4, 2006, pp. 826-834.
[32] A. Kramer, I. Schwebke and G. Kampf, “How Long do Nosocomial Pathogens Persist on Inanimate Surfaces? A Systematic Review,” BMC Infectious Diseases, Vol. 6, 2006, p. 130. doi:10.1186/1471-2334-6-130
[33] R. B. Tompkin, et al., “Guidelines to Prevent Post-Processing Contamination from Listeria monocytogenes,” Dairy, Food and Environmental Sanitation, Vol. 19, No. 8, 1999, pp. 551-562.
[34] H. Takahashi, et al., “Desiccation Survival of Listeria monocytogenes and Other Potential Foodborne Pathogens on Stainless Steel Surfaces is Affected by Different Food Soils,” Food Control, Vol. 22, No. 3-4, 2011, pp. 633-637. doi:10.1016/j.foodcont.2010.09.003
[35] J. W. Costerton, et al., “Microbial Biofilms,” Annual Review of Microbiology, Vol. 49, 1995, pp. 711-742. doi:10.1146/annurev.mi.49.100195.003431
[36] J. D. Stopforth, et al., “Biofilm Formation by Acid-Adapted and Non-Adapted Listeria monocytogenes in Fresh Beef Decontamination Washings and Its Subsequent Inactivation with Sanitizers,” Journal of Food Protection, Vol. 65, No. 11, 2002, pp. 1717-1727.
[37] G. McDonnell and A. D. Russell, “Antiseptics and Disinfectants: Activity, Action, and Resistance,” Clinical Microbiology Reviews, Vol. 14, No. 1, 2001, pp. 227-228.
[38] E. Ortega Morente, et al., “Biocide Tolerance in Bacteria,” International Journal of Food Microbiology, Vol. 162, No. 1, 2013, pp. 13-25. doi:10.1016/j.ijfoodmicro.2012.12.028
[39] S. Langsrud and G. Sundheim, “Factors Contributing to the Survival of Poultry Associated Pseudomonas spp. Exposed to a Quaternary Ammonium Compound,” Journal of Applied Bacteriology, Vol. 82, No. 6, 1997, pp. 705-712. doi:10.1046/j.1365-2672.1997.00186.x
[40] E. Heir, G. Sundheim and A. Holck, “Identification and Characterization of Quaternary Ammonium Compound Resistant Staphylococci from the Food Industry,” International Journal of Food Microbiology, Vol. 48, No. 3, 1999, pp. 211-219. doi:10.1016/S0168-1605(99)00044-6
[41] E. Heir, G. Sundheim and A. L. Holck, “Resistance to Quaternary Ammonium Compounds in Staphylococcus spp. Isolated from the Food Industry and Nucleotide Sequence of the Resistance Plasmid pST827,” Journal of Applied Bacteriology, Vol. 79, No. 2, 1995, pp. 149-156. doi:10.1111/j.1365-2672.1995.tb00928.x
[42] D. O. Kolawole, “Resistance Mechanisms of Mucoid-Grown Staphylococcus aureus to the Antibacterial Action of Some Disinfectants and Antiseptics,” FEMS Microbiology Letters, Vol. 25, No. 2-3, 1984, pp. 205-209. doi:10.1111/j.1574-6968.1984.tb01457.x
[43] S. Langsrud, T. Moretro and G. Sundheim, “Characterization of Serratia marcescens Surviving in Disinfecting Footbaths,” Journal of Applied Microbiology, Vol. 95, No. 1, 2003, pp. 186-195. doi:10.1046/j.1365-2672.2003.01968.x
[44] H. Maseda, et al., “Mutation in the sdeS Gene Promotes Expression of the sdeAB Efflux Pump Genes and Multidrug Resistance in Serratia marcescens,” Antimicrobial Agents and Chemotherapy, Vol. 55, No. 6, 2011, pp. 2922-2926. doi:10.1128/AAC.01755-10
[45] L. Guillier, et al., “Modelling the Competitive Growth between Listeria monocytogenes and Biofilm Microflora of Smear Cheese Wooden Shelves,” International Journal of Food Microbiology, Vol. 128, No. 1, 2008, pp. 51-57. doi:10.1016/j.ijfoodmicro.2008.06.028

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