Control of Staphylococcus aureus in Meat System by in situ and ex situ Bacteriocins from Lactobacillus sakei and Pediococcus spp.

Authors: M. Naimi, M. B. Khaled

Abstract:

The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physico-chemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: Biocontrol, in situ and ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1093118

References:

[1] U. Schillinger, and F.K. Lücke, "Identification of lactobacilli from meat and meat products”, Food Microbiology.4, 1987, 199-208.
[2] Y. Morishita, and K. Shiromizu, "Characterization of lactobacilli isolated from meats and meat products”, International Journal of Food Microbiology, 3, 1986, 19-29.
[3] J. Samelis, F. Maurogenakis, and J. Metaxopoulos, "Characterisation of lactic acid bacteria isolated from naturally fermented Greek dry salami”, International Journal of Food Microbiology, 23, 1994, 179-196.
[4] A. Najjari, H. Ouzari, A. Boudabous, and M. Zagorec, "Method for reliable isolation of Lactobacillus sakei strains originati from Tunisian seafood and meat products”, International Journal of Food Microbiology, 121, 2008, 342–351.
[5] R. Bromberg, I. Moreno, C.L. Zaganini, R.R. Delboni, and J. De Oliveira, "Isolation of bacteriocin-producing lactic acid bacteria from meat and meat products and its spectrum of inhibitory activity”, Brazilian Journal of Microbiology, Vol.35, 2004.
[6] R. J. Jones, H. M. Hussein, M. Zagorec, G. Brightwell, and J. R Tagg, "Isolation of lactic acid bacteria with inhibitory activity against pathogens and spoilage organisms associated with fresh meat”, Food Microbiology, Vol. 25, 2008.
[7] E. C. P. De-Martinis, and F .Z. Freitas, "Screening of lactic acid bacteria from Brazilian meats for bacteriocin formation”, Food Control, Vol. 14, 2003.
[8] M. A. H. Al-Allaf, A. M. M. Al-Rawi, and A. T. Al-Mola, "Antimicrobial activity of lactic acid bacteria isolated from minced beef meat against some pathogenic bacteria”, Iraqi Journal of Veterinary Sciences. Vol 23 Supplement I (2009) 115-117.
[9] J. Chaiyana, S. Boonrang, and S. Sinsuwongwat, "Isolation and screening of bactériocin producing bacteria from fermented meat products”, Biotechnology for gross national hapipiness, 2007, 245- 258.
[10] P. H. Castellano, W. H. Holzapfel,andG. M. Vignolo, "The control of Listeria innocua and Lactobacillus sakei in broth and meat slurry with the bacteriocinogenic strain Lactobacillus casei CRL705”, Food Microbiology, 21, 2004, 291-298.
[11] C. Charlier, M. Cretenet, S. Even, and Y. Le Loir, "Interactions between Staphylococcus aureus and lactic acid bacteria: An old story with new perspectives”, International Journal of Food Microbiology. 13, 2009 30–39.
[12] A. Gálvez, H. Abriouel, R. L. López, and N. Ben Omar, "Bacteriocinbased strategies for food biopreservation”, International Journal of Food Microbiology. 120, 2007, 51-70.
[13] T. R. Klaenhammer, T. R. Barrangou, R. Buck, B. L. Azcarate-Peril, and M. A. Altermann, "Genomic features of lactic acid bacteria effecting bioprocessing and health”, FEMS Microbiol Rev, Vol. 29, 2005.
[14] P. Castellano, C. Belfiore, S. Fadda, and G. Vignolo, "A review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina”, Meat Science, Vol. 79, 2008.
[15] C. Dortu, and P. Thonart, "Les bactériocines des bactéries lactiques: caractéristiques et intérêts pour la bioconservation des produits alimentaires”, Biotechnol. Agron. Soc. Environ. 13, 2009.
[16] F. K Lucke, "Utilization of microbes to process and preserve meat”, Meat Science, Vol.56, 2000, 105–115.
[17] W. H. Holzapfel, R. Geisen, and U. Schillinger, "Biological preservation of foods with reference to protective cultures, bacteriocins and foodgrade enzymes”, International Journal of Food Microbiology. 24, 1995, 343–36.
[18] T.R. Klaenhammer, "Bacteriocins of lactic acid bacteria”, Biochimie, Vol. 70, 1988.
[19] T. Abee, L. Krockel, and C. Hill, "Bacteriocins : modes of action and potentials in food preservation and control of food poisoning”, International Journal of Food Microbiology, Vol. 28, 1995.
[20] A. S Naidu, R. Unal, and J. Tulpinski, "Bacteriocins: Antimicrobial Activity and Applications”, In: K. Shetty, G. Paliyath, Anthony, Pometto, and R. E. Levin, food biotechnology, 2006.
[21] T.R Klaenhammer, "Genetics of bacteriocins produced by lactic acid bacteria”, Federation of European Microbiological Societies, Microbiology Reviews, Vol. 12 1993.
[22] L. H. Deegana, P. D. Cottera, and C. Hilla, "Bacteriocine biological tools for biopreservation”, International Dairy Journal, Vol.16, 2006, 1058–1071.
[23] M. Hugas, "Bacteriocinogenic Lactic Acid Bacteria for the Biopreservation of Meat and Meat Products”, Meat Science. Vol 49, 1998, 139-150
[24] J. Guiraud, and P. Galzy, "L’analyse Microbiologique dans les industries alimentaires”, L’usine, 1980, P235.
[25] M. E. Stiles, "Potential for biological control of agents of foodborne disease”, Food Research International, Vol. 27, 1994.
[26] B. Ray, and A. bhunia, "Fundamental Food Microbiology”, 4 th edition, CRC Press, 2008, 492 pp.
[27] I. Kalalou, M. Faid, and A. T. Ahami, "Extending shelf life of fresh minced camel meat at ambient temperature by Lactobacillus dlbrueckii subsp. Delbrueckii”, Electronic Journal of Biotechnology, Vol.7, 2004.
[28] S. Ananou, M. Maqueda, M. Martínez-Bueno, A. Gálvez, and E. Valdivia, "Control of Staphylococcus aureus in sausages by enterocin AS-48”, Meat Science. 71, 2005, 549–556.
[29] H. Khan, S. Flint, and P. L. Yu, "Enterocins in food preservation, International Journal of Food Microbiology”, International Journal of Food Microbiology, 2010.
[30] S. Ananou, M. Garriga, A. Jofré, T. Aymerich, A. Gálvez, M. Maqueda, M. Martínez-Bueno, and E. Valdivia, "Combined effect of enterocin AS- 48 and high hydrostatic pressure to control food-borne pathogens inoculated in low acid fermented sausages”, Meat Science, 2009.
[31] S. Ananou, M. Garriga, M. Hugas, M. Maqueda, M. Martínez-Bueno, A. Gálvez, and E. Valdivia, "Control of Listeria monocytogenes in model sausages by enterocin AS-48”, International Journal of Food Microbiology, Vol.103, 2005, 179-190.
[32] L. V. Thomas, M. R. Clarkson, and J. Delves-Broughton, "Nisin”, in: A. S. Naidu, Natural food antimicrobial systems, Boca Raton: CRC Pres, 2000, 463– 524.
[33] G.F. De-Valdez, "Maintenance of Lactic Acid Bacteria”,in: J. F. T. Spencer, and A. L. R. Spencer, Food Microbiology protocols, Humana Press, 2001.
[34] A. Badis, N. Laouabdia-Sellami, D. Guetarni, M. Kihal, and R. Ouzrout, "caracterisationphenotypique des bacteries lactiques isolees à partir de lait cru de chèvre de deux populations caprines locales "arabia et kabyle"”, Sciences et Technologie, Vol. 23, 2005.
[35] L. J. Harris, M. A. Daeschel, M. E. Stiles, and T. R. Klaenhammer, "Antimicrobial activity of lactic acid bacteria against listeria monocytogenes”, Journal of Food Protection, Vol. 52, 1989.
[36] U. Schillinger, and F.K. Lücke, "Antibacterial Activity of Lactobacillus sake Isolated from Meat”, applied and environmental Microbiology, Vol. 55, 1989.
[37] S. A. Cuozzo, J. M. Sesma-Fernando, A. A. Holgado, R. Pesce, and R. R. Raya, "Methods for the Detection and Concentration of Bacteriocins Produced by Lactic Acid Bacteria”,in: F. T. S. John, and L .R. S. Alicia, "Food Microbiology protocols”, Humana Press, 2001.
[38] A. Delgado, D. Brito, P. Fevereiro, R. Tenreiro, and C. Peres, "Bioactivity quantification of crude bacteriocin solutions”, Journal of Microbiological Methods, Vol. 62, 2005.
[39] G. Greer, and D. Dilts-Bryan, "Lactic acid inhibition of the growth of spoilage bacteria and cold tolerant pathogens on pork”, International Journal of Food Microbiology. 25, 1995, 141-151.
[40] J. Cleveland, T. J. Montville, I. F. Nes, and M. L. Chikindas, "Bacteriocins: safe, natural antimicrobials for food preservation”, International Journal of Food Microbiology. 71, 2001, 1–20.
[41] A. L. Vigil, E. Palou, M. E. Parish, and D. P. Michael, "Methods for Activity Assay and Evaluation of Results”,in: D. P. Michael, J. N. Sofos, and A. L. Branen, Antimicrobials in food , 2005, Third Edition, CRC Press : 659- 680.
[42] Y. Babji, and T. R. K. Murthy, "Effect of inoculation of mesophilic lactic acid bacteria on microbial and sensory changes of minced goat meat during storage under vacuum and subsequent aerobic storage”, Meat Science, 54, 2000, 197- 202.