Potential of cell-free supernatants from cultures of selected lactic acid bacteria and yeast obtained from local fermented foods as inhibitors of Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus

BMC Research Notes

Abraham Aseffa²  Nigatu Endalafer²  Emawayish Andargie²  Tewodros Tariku²  Nigus Zegeye¹  Solomon H Mariam² 

[1] Debre Berhan University, Debre Berhan, Ethiopia;Armauer Hansen Research Institute, P.O. Box 1005, Addis Ababa, Ethiopia

关键词: Inhibition;    Cell-free supernatant;    Inhibition;    Staphylococcus aureus;    Salmonella;    Listeria monocytogenes;    Lactic acid bacteria;   

Others  :  1129824
DOI  :  10.1186/1756-0500-7-606

 received in 2014-08-02, accepted in 2014-08-11,  发布年份 2014

【 摘 要 】

Background

Food-borne infections cause huge economic and human life losses worldwide. The most common contaminants of foods include Listeria monocytogenes Salmonellae and Staphylococcus aureus. L. monocytogenes is most notorious due to its tolerance to common food preservation methods and the risks it poses, including higher fatality rates. Safer, more efficacious control methods are thus needed. Along with food-borne pathogens, lactic acid bacteria (LAB) can also be found in foods. Some LAB isolates inhibit pathogenic bacteria by various mechanisms, including by production of antimicrobial metabolites.

Methods

The potential of cell-free culture supernatants (CFS) derived from broth cultures of selected local LAB and yeast isolates, some of which were subjected to various treatments, were tested for inhibition of L. monocytogenes, Salmonella spp. and S. aureus in in vitro culture by incorporating various proportions of the CFSs into the growth medium concurrently with inoculation (co-cultures) or following limited proliferation after inoculation of the pathogens (delayed cultures). The effects of the CFSs on various growth parameters were assessed.

Results

CFS from the LAB isolates were strongly inhibitory when co-cultured. The inhibitory activities were stable following heat or protease treatment of the CFSs. Inhibitory activity was dependent primarily on active substance(s) secreted into the supernatant. In all co-cultures, CFS proportion-dependent progressive decrease in the number of colonies was observed and both growth rates and number of generations were reduced with significantly fewer numbers of colony forming units, whereas generation times were significantly increased compared to those of controls. Transfer from co-cultures to fresh broth showed inhibited cultures contained bacteria that can re-grow, indicating the presence of viable bacteria that are undetectable by culture. Growth rates in CFS-treated delayed cultures were also reduced to varying degrees with the number of colonies in some cultures being significantly less than the corresponding control values. CFSs were active against both Gram-positive and –negative bacteria.

Conclusions

Active metabolites produced and secreted by LAB into the growth medium were effective in inhibiting the tested pathogens. Early addition of the CFSs was necessary for significant inhibition to occur. Further studies will help make these findings applicable to food safety.

【 授权许可】

   
2014 Mariam et al.; licensee BioMed Central Ltd.

【 参考文献 】

• [1]Hoffmann S, Batz MB, Morris JG: Annual cost of illness and quality-adjusted life year losses in the United States due to 14 foodborne pathogens. J Food Prot 2012, 75:1292-1302.
• [2]Scharff RL: Economic burden from health losses due to foodborne illness in the United States. J Food Prot 2012, 75:123-131.
• [3]Food Safety: Centers for disease control and prevention. http://www.cdc.gov/foodsafety/facts.html webcite
• [4]Pan Y, Breidt F, Kathariou S: Resistance of Listeria monocytogenes biofilms to sanitizing agents in a simulated food processing environment. Appl Environ Microbiol 2006, 72:7711-7717.
• [5]Müller A, Rychli K, Muhterem-Uyar M, Zaiser A, Stessl B, Guinane CM, Cotter PD, Wagner M, Schmitz-Esser S: Tn6188 - a novel transposon in Listeria monocytogenes responsible for tolerance to benzalkonium chloride. PLoS One 2013, 8:e76835.
• [6]Vázquez-Boland JA, Kuhn M, Berche P, Chakraborty T, Domínguez-Bernal G, Goebel W, González-Zorn B, Wehland J, Kreft J: Listeria pathogenesis and molecular virulence determinants. Clin Microbiol Rev 2001, 14:584-640.
• [7]Camejo A, Carvalho F, Reis O, Elsa Leitão E, Sousa S, Cabanes D: The arsenal of virulence factors deployed by Listeria monocytogenes to promote its cell infection cycle. Virulence 2011, 2:379-394.
• [8]Abee T, Krockel L, Hill C: Bacteriocins: modes of action and potentials in food preservation and control of food poisoning. Int J Food Microbiol 1995, 28:169-185.
• [9]Caplicec E, Fitzgerald GF: Food fermentations: role of microorganisms in food production and preservation. Int J Food Microbiol 1999, 50:131-149.
• [10]Natrajan N, Sheldon BW: Inhibition of Salmonella on poultry skin using protein- and polysaccharide-based films containing a nisin formulation. J Food Prot 2000, 63:1268-1272.
• [11]Benkerroum N, Oubel H, Sandine WE: Effect of nisin on yogurt starter, and on growth and survival of Listeria monocytogenes during fermentation and storage of yogurt. Int J Food Saf 2003, 1:1-5.
• [12]De Vuyst L, Leroy F: Bacteriocins from lactic acid bacteria: production, purification, and food applications. J Mol Microbiol Biotechnol 2007, 13:194-199.
• [13]Atassi F, Servin AL: Individual and co-operative roles of lactic acid and hydrogen peroxide in the killing activity of enteric strain Lactobacillus johnsonii NCC933 and vaginal strain Lactobacillus gasseri KS120.1 against enteric, uropathogenic and vaginosis-associated pathogens. FEMS Microbiol Lett 2010, 304:29-38.
• [14]O’Shea EF, Cotter PD, Ross RP, Hill C: Strategies to improve the bacteriocin protection provided by lactic acid bacteria. Curr Opin Biotechnol 2013, 24:130-134.
• [15]Dabour N, Zihler A, Kheadr E, Lacroix C, Fliss I: In vivo study on the effectiveness of pediocin PA-1 and Pediococcus acidilactici UL5 at inhibiting Listeria monocytogenes. Int J Food Microbiol 2009, 133:225-233.
• [16]Riley MA, Robinson SM, Roy CM, Dennis M, Liu V, Dorit RL: Resistance is futile: the bacteriocin model for addressing the antibiotic resistance challenge. Biochem Soc Trans 2012, 40:1438-1442.
• [17]Goh Y-L, He H, March JC: Engineering commensal bacteria for prophylaxis against infection. Curr Opin Biotechnol 2012, 23:924-930.
• [18]Wells J: Mucosal vaccination and therapy with genetically modified lactic acid bacteria. Ann Rev Food Sci Technol 2011, 2:423-445.
• [19]Koo OK, Amalaradjou MAR, Bhunia AK: Recombinant probiotic expressing Listeria adhesion protein attenuates Listeria monocytogenes virulence in vitro. PLoS One 2012, 7:e29277.
• [20]Cotter PD, Ross RP, Hill C: Bacteriocins - a viable alternative to antibiotics? Nat Rev Microbiol 2013, 11:95-105.
• [21]Joint Food and Agriculture Organization of the United Nations/World Health Organization: Working Group Report on Drafting Guidelines for the Evaluation of Probiotics in Food, London, Ontario, Canada. April 30 and May, 2002 [cited 2010 Aug 25]. ftp://ftp.fao.org/es/esn/food/wgreport2.pdf
• [22]MacFadden RR: Biochemical Tests for Identification of Medical Bacteria. 3rd edition. Philadelphia: Lippincott Williams and Wilkins; 2000.
• [23]Murray PR, Shea Y: Guide to Clinical Microbiology. 3rd edition. Washington, DC: American Society for Microbiology; 2004.
• [24]Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory: New York; 1989.
• [25]Cleveland J, Montville TJ, Nes IF, Chikindas ML: Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 2001, 71:1-20.
• [26]Cotter PD, Hill C, Ross RP: BACTERIOCINS: developing innate immunity for food. Nat Rev Microbiol 2005, 3:777-788.
• [27]Jack RW, Tagg JR, Ray B: Bacteriocins of gram-positive bacteria. Microbiol Rev 1995, 59:171-200.
• [28]Mélançon D, Grenier D: Production and properties of bacteriocin-like inhibitory substances from the swine pathogen Streptococcus suis serotype 2. Appl Environ Microbiol 2003, 69:4482-4488.
• [29]Walls T, Power D, Tagg J: Bacteriocin-like inhibitory substance (BLIS) production by the normal flora of the nasopharynx: potential to protect against otitis media? J Med Microbiol 2003, 52:829-833.
• [30]Gálvez A, Abriouel H, Benomar N, Lucas R: Microbial antagonists to food-borne pathogens and biocontrol. Curr Opin Biotechnol 2010, 21:142-148.
• [31]Rodríguez JM, Martínez MI, Suárez AM, Martínez JM, Hernández PE: Research note: unsuitability of the MRS medium for the screening of hydrogen peroxide-producing lactic acid bacteria. Lett Appl Microbiol 1997, 25:73-74.
• [32]Pridmore RD, Pittet AC, Praplan F, Cavadini C: Hydrogen peroxide production by Lactobacillus johnsonii NCC 533 and its role in anti-Salmonella activity. FEMS Microbiol Lett 2008, 283:210-215.
• [33]Bleicher A, Stark T, Hofmann T, Bogovic Matijasić B, Rogelj I, Scherer S, Neuhaus K: Potent antilisterial cell-free supernatants produced by complex red-smear cheese microbial consortia. J Dairy Sci 2010, 93:4497-4505.
• [34]Bouttefroy A, Millie’re J-B: Nisin–curvaticin 13 combinations for avoiding the regrowth of bacteriocin resistant cells of Listeria monocytogenes ATCC 15313. Int J Food Microbiol 2000, 62:65-75.
• [35]Hartmann HA, Wilke T, Erdmann R: Efficacy of bacteriocin-containing cell-free culture supernatants from lactic acid bacteria to control Listeria monocytogenes in food. Int J Food Microbiol 2011, 146:192-199.