Document Type : Original Article

Authors

1 DVM Graduated student, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

Abstract

This study was carried out to investigate the stability, antibacterial properties and biofilm removal potential of cell-free supernatant (CFS) of Lactobacillus acidophilus LA5 and Lactobacillus casei 431 against Staphylococcus aureus ATCC 25923. Antibacterial activity of both Lactobacillus strains was measured according to the agar spot method. The CFS was prepared by centrifugation of bacterial suspension at 4000 g for 10 min and the antimicrobial activity was measured using agar-well diffusion. The stability of CFSs during storage at 4.00 ± 2.00 °C and 25.00 ± 2.00 °C for a period of 4 weeks was measured based on the method of broth micro-dilution assay. Moreover, biofilm removal potential of CFS on 2-days-old biofilm of S. aureus­ developed on polystyrene and glass surfaces was also determined. The efficacy of CFS on bacterial biofilm established on the glass surface was also observed using fluorescence microscope. Results showed that inhibition zones of L. acidophilus (50.26 mm) were greater than L. casei (37.06 mm). The minimum inhibitory concentration of both CFSs remained stable (40 mg mL-1) during the storage for 28 days at 4.00 and 25.00 °C and storage temperature did not affect the antibacterial effectiveness of CFS. The addition of both CFSs significantly removed biofilm developed on both tested surfaces in a concentration-dependent manner. Biofilm removal property of L. acidophilus CFS was generally better than L. casei CFS which was confirmed by fluorescence microscope. The application of CFS of probiotic strains (i.e. Lactobacillus) as antibacterial and biofilm removal compounds could be very suitable to control the growth of food-borne pathogens.

Keywords

Main Subjects

  1. Lee JS, Bae YM, Lee SY, et al. Biofilm formation of Staphylococcus aureus on various surfaces and their resistance to chlorine sanitizer. J Food Sci 2015; 80(10): M2279-M2286.
  2. Kadariya J, Smith TC, Thapaliya D. Staphylococcus aureus and staphylococcal food-borne disease: an ongoing challenge in public health. BioMed Res Int 2014; 2014: doi: 10.1155/2014/827965.
  3. Méndez-Vilas A. Microbial pathogens and strategies for combating them: Science, Technology and Education. Badajoz, Spain: Formatex 2013: 42-51.
  4. Khiralla GM, Mohamad EAH, Farag AG, et al. Antibiofilm effect of Lactobacillus pentosus and Lactobacillus plantarum cell-free supernatants against some bacterial pathogens. J Biotech Res 2015; 6: 86-95.
  5. Shunmugaperumal T. Biofilm eradication and prevention: A pharmaceutical approach to medical device infections. Hoboken, USA: John Wiley & Sons 2010; 3-36.
  6. Srey S, Jahid IK, Ha SD. Biofilm formation in food industries: A food safety concern. Food Control 2013; 31(2): 572-585.
  7. Qing L, Xiaoli L, Mingsheng D, et al. Aggregation and adhesion abilities of 18 lactic acid bacteria strains isolated from traditional fermented food. Int J Agri Pol Res 2015; 3(2): 84-92.
  8. Ait Ouali F, Al Kassaa I, Cudennec B, et al. Identification of lactobacilli with inhibitory effect on biofilm formation by pathogenic bacteria on stainless steel surfaces. Int J Food Microbiol 2014; 191: 116-124.
  9. Jalilsood T, Baradaran A, Song AAL, et al. Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: A potential host for the expression of heterologous proteins. Microb Cell Fact 2015; 14: 96. doi: 10.1186/s12934-015-0283-8.
  10. Gomez NC, Ramiro JM, Quecan BX, et al. Use of potential probiotic lactic acid bacteria (lab) biofilms for the control of Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7 biofilms formation. Front Microbiol 2016; 7: 863. doi: 10.3389/fmicb.2016.00863.
  11. Aminnezhad S, Kasra-Kermanshahi R. Antibiofilm activity of cell-free supernatant from Lactobacillus casei in Pseudomonas aeruginosa. Feyz 2014; 18(1): 30-37.
  12. Bulgasem BY, Hassan Z, Abdalsadiq NK, et al. Anti-adhesion activity of lactic acid bacteria supernatant against human pathogenic Candida species biofilm. Health Sci J 2015; 9(6): 3.
  13. Wang HH, Ye KP, Zhang QQ, et al. Biofilm formation of meat-borne Salmonella enterica and inhibition by the cell-free supernatant from Pseudomonas aeruginosa. Food Control 2013; 32(2): 650-658.
  14. Awaisheh SS, Ibrahim SA. Screening of antibacterial activity of lactic acid bacteria against different pathogens found in vacuum-packaged meat products. Foodborne Pathog Dis 2009; 6(9): 1125-1132.
  15. Marques JDL, Funck GD, Dannenberg GDS, et al. Bacteriocin-like substances of Lactobacillus curvatus P99: characterization and application in biodegradable films for control of Listeria monocytogenes in cheese. Food Microbiol 2017; 63: 159-163.
  16. Devi Avaiyarasi N, David Ravindran A, Venkatesh P, et al. In vitro selection, characterization and cytotoxic effect of bacteriocin of Lactobacillus sakei GM3 isolated from goat milk. Food Control 2016; 69: 124-133.
  17. CLSI. Clinical and Laboratory Standards Institute: performance standards for antimicrobial susceptibility testing; twentieth informational supplement. Document M100-S22. Wayne, USA; CLSI 2012.
  18. Mahdavi M, Jalali M, Kasra Kermanshahi R. The effect of nisin on biofilm forming foodborne bacteria using micro-titer plate method. Res Pharm Sci 2007; 2(2): 113-118.
  19. Valeriano C, de Oliveira TLC, de Carvalho SM, et al. The sanitizing action of essential oil-based solutions against Salmonella enterica serotype Enteritidis S64 biofilm formation on AISI 304 stainless steel. Food Control 2012; 25(2): 673-677.
  20. Perez Ibarreche M, Castellano P, Vignolo G. Evaluation of anti-Listeria meat borne Lactobacillus for biofilm formation on selected abiotic surfaces. Meat Sci 2014; 96(1): 295-303.
  21. Totoonchi P, Hesari J, Moradi M, et al. Production and evaluation of probiotic red grape juice by Latobacillus acidophilus LA5, and Lactobacillus casei. J Food Sci Res 2015; 25(4): 655-666
  22. Mirnejad R, Vahdati AR, Rashidiani J, et al. The antimicrobial effect of lactobacillus casei culture supernatant against multiple drug resistant clinical isolates of Shigella sonnei and Shigella flexneri in vitro. Iran Red Crescent Med J 2013; 15(2): 122-126.
  23. Sharma C, Singh BP, Thakur N, et al. Antibacterial effects of Lactobacillus isolates of curd and human milk origin against food-borne and human pathogens. Biotech 2017; 7(1): 31.
  24. Mahasneh AM, Hamdan S, Mahasneh SA. Probiotic properties of Lactobacillus species isolated from local traditional fermented products. Jordan J Biol Sci 2015; 8(2): 81-87.
  25. Tabasco R, García-Cayuela T, Peláez C, et al. Lactobacillus acidophilus La-5 increases lactacin B production when it senses live target bacteria. Int J Food Microbiol 2009; 132(2-3): 109-116.
  26. Abedi D, Feizizadeh S, Akbari V, et al. In vitro anti-bacterial and anti-adherence effects of Lactobacillus delbrueckii subsp bulgaricus on Escherichia coli. Res Pharm Sci 2013; 8(4): 260-268.
  27. Valéria Garcia Pereira VG, Gómez RJH. Antimicrobial activity of Lactobacillus acidophilus against foodborne pathogens. Semina Ciencias Agrarias2007; 28(2): 229-239.
  28. Ehrmann MA, Kurzak P, Bauer J, et al. Characterization of lactobacilli towards their use as probiotic adjuncts in poultry. J Appl Microbiol 2002; 92(5): 966-975.
  29. Vuotto C, Barbanti F, Mastrantonio P, et al. Lactobacillus brevis CD2 inhibits Prevotella melaninogenica biofilm. Oral Dis 2014; 20(7): 668-674.
  30. Kim Y, Oh S, Kim SH. Released exopolysaccharide (r-EPS) produced from probiotic bacteria reduce biofilm formation of enterohemorrhagic Escherichia coli O157:H7. Biochem Biophys Res Commun 2009; 379(2): 324-329.
  31. Ciandrini E, Campana R, Casettari L, et al. Characterization of biosurfactants produced by Lactobacillus spp. and their activity against oral Streptococci biofilm. Appl Microbiol Biotechnol 2016; 100(15): 6767-6777.
  32. Auger S, Ramarao N, Faille C, et al. Biofilm formation and cell surface properties among pathogenic and nonpathogenic strains of the Bacillus cereus group. Appl Environ Microbiol 2009; 75(20): 6616-6618.