Sevim Feyza ERDOĞMUŞ, Gül DURMAZ, Zahide KÖSE, Beytullah KENAR, Mine ERİK, Safiye Elif KORCAN

The determination of antimicrobial and antibiofilm activities of foodborne lactic acid bacteria against Enterobacter cloacae isolates

The aim of this study was to investigate the antimicrobial and antibiofilm activities of 10 different lactic acid bacteria (LAB)strains isolated from local food sources of animal origin against 4 Enterobacter cloacae isolates obtained from clinical cases and determinetheir adhesion potentials to intestinal epithelial cells. In this study, all Enterobacter cloacae isolates (P3, P4, P5, P7) identified with theBD Phoenix automation system were detected to form biofilm with both Congo red agar and Microtiter plate methods. Amoxicillinclavulanate,cefuroxime, and ampicillin resistance was determined in all isolates. It was determined that LAB strains producingexopolysaccharide (EPS) were able to colonize intestinal epithelial cells. It is noteworthy that LAB extracts were effective to inhibitthe biofilm formation of P3Ec, which had higher antibiotic resistance than those of other isolates. Antimicrobial effect of LAB extractson Enterobacter cloacae were also detected by both agar disc diffusion and well diffusion tests. In this study, all of the isolated LABstrains (especially L. lactis, L. fermentum, and L. casei) are good candidates for controlling Enterobacter cloacae biofilm formation. Thesefindings indicate that L. lactis, L. fermentum, and L. casei can potentially be developed as novel antibiofilm agents.

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1. Cramton SE, Gerke C, Schnell NF, Nichols WW, Gotz F. The intercellular adhesion [ica] locus is present in Staphylococcus aureus and is required for biofilm formation. Infection and Immunity 1999; 67: 5427-33.
2. Roy R, Tiwaria M, Donellib G, Tiwari V. Strategies for combating bacterial biofilms: a focus on anti-biofilm agents and their mechanisms of action. Virulence 2018; 9 (1): 522- 554.
3. Rasmussen TB, Givskov M. Quorum sensing inhibitors: a bargain of effects. Microbiology 2006; 152 (4): 895-904.
4. Soares GG, Costa JF, Melo Flávia BS, Mola R, Balbino TCL. Biofilm production and resistance profile of Enterobacter spp. strains isolated from pressure ulcers in Petrolina, Pernambuco, Brazil. Jornal Brasileiro de Patologia e Medicina Laboratorial 2016; 52 (5): 293-298.
5. Sarowska J, Choroszy-Król I, Regulska-Ilow B, Frej-Madrzak M, Jama-Kmiecik A. The therapeutic effect of probiotic bacteria on gastrointestinal diseases. Advances in Clinical and Experimental Medicine 2013; 22 (5): 759-766.
6. Stewart PS, Costerton JW. Antibiotic resistance of bacteria in biofilms. Lancet 2001; 358 (9276): 135-138.
7. Cui X, Shi Y, Gu S, Yan X, Chen H et al. Antibacterial and antibiofilm activity of lactic acid bacteria isolated from traditional artisanal milk cheese from northeast China against enteropathogenic bacteria. Probiotics and Antimicrobial Proteins 2018; 10 (4): 601-610.
8. De Roock S, Van Elk M, Van Dijk ME, Timmerman HM, Rijkers GT et al. Lactic acid bacteria differ in their ability to induce functional regulatory T cells in humans. Clinical and Experimental Allergy 2010; 40 (1): 103-110.
9. Corthésy B, Gaskins HR, Mercenier A. Cross-talk between probiotic bacteria and the host immune system. Journal of Nutrition 2007; 137 (3): 781-790.
10. Servin AL. Antagonistic activities of lactobacilli and bifidobacteria against microbial pathogens. FEMS Microbiology Reviews 2004; 28 (4): 405-440.
11. Freeman DJ, Falkiner FR, Keane CT. New method for detecting slime production by coagulase negative staphylococci. Journal of Clinic Pathology 1989; 42: 872-874.
12. Jain A, Agarwal A. Biofilm production, a marker of pathogenic potential of colonizing and commensal staphylococci. Journal of Microbiological Methods 2009; 76: 88-92.
13. Şahin R. Staphylococcus aureus suşlarında biyofilm üretimi, biyofilm pozitif ve negatif suşların genotipik ve fenotipik karakterlerinin karşılaştırılması. Thesis, Pamukkale University, Denizli, Turkey, 2007 (in Turkish).
14. Sandberg M, Maattanen A, Peltonen J, Vuorela PM, Fallarero A. Automating a 96-well microtitre plate model for Staphylococcus aureus biofilms: an approach to screening of natural antimicrobial compounds. International Journal of Antimicrobial Agents 2008; 32: 233-240.
15. Rashid MH, Kornberg A. Inorganic polyphosphate is needed for swimming, swarming, and twitching motilities of Pseudomonas aeruginosa. Proceedings of the National Academy of Sciences of the USA 2000; 97: 4885-4890.
16. Deziel E, Comeau Y, Villemur R. Initiation of biofilm formation by P. aeruginosa 57RP correlates with emergence of hyperpiliated and highly adherent phenotypic variants deficient in swimming, swarming, and twitching motilities. Journal of Bacteriology 2001; 183: 1195-1204.
17. Marshall VM, Rawson HL. Effects of exopolysaccharide producing strains of thermophilic lactic acid bacteria on texture of stirred yoghurt. International Journal of Food Science & Technology 1999; 34: 137-143
18. Pringsulaka O, Thongngam N, Suwannasai N, Atthakor W, Pothıvejkul K et al. Partial characterisation of bacteriocins produced by lactic acid bacteria isolated from Thai fermented meat and fish. Food Control 2012; 23: 547-551.
19. Schillinger U, Luke FK. Antibacterial activity of Lactobacillus sake isolated from meat. Applied and Environmental Microbiology 1989; 55 (8): 1901-1906.
20. Thenmozhi R, Nithyanand P, Rathna J, Karutha Pandian S. Antibiofilm activity of coral- associated bacteria against different clinical M serotypes of Streptococcus pyogenes. FEMS Immunology Medical Microbiology 2009; 57: 284-294.
21. Nithyanand P, Pandian SK. Phylogenetic characterization of culturable bacterial diversity associated with the mucus and tissue of the coral Acropora digitifera from Gulf of Mannar. FEMS Microbiology Ecology 2009; 69: 384-394.
22. Theelen MJ, Wilson WD, Edman JM, Magdesian KG, Kass PH. Temporal trends in prevalence of bacteria isolated from foals with sepsis: 1979–2010. Equine Veterinary Journal 2014; 46: 169-173.
23. Adams DJ, Eberly MD, Goudie A, Nylund CM. Rising vancomycin resistant enterococcus infections in hospitalized children in the United States. Hospital Pediatrics 2016; 6: 404- 411.
24. Arias CA, Contreras GA, Murray BE. Management of multidrug-resistant enterococcal infections. Clinical Microbiology and Infection 2010; 16: 555-562.
25. Devriese LA, Laurier L, Herdt PD, Haesebrouck F. Enterococcal and streptococcal species isolated from faeces of calves, young cattle and dairy cows. Journal of Applied Bacteriology 1992; 72: 29-31.
26. Bradley AJ, Leach KA, Breen JE, Green LE, Green MJ. Survey of the incidence and aetiology of mastitis on dairy farms in England and Wales. Veterinary Record 2007; 160: 253-257.
27. Nam HM, Lim SK, Moon JS, Kang HM, Kim JM et al. Antimicrobial resistance of Enterococci isolated from mastitic bovine milk samples in Korea. Zoonoses and Public Health 2010; 57 (7-8): 59-64.
28. Wu X, Hou S, Zhang Q, Ma Y, Zhang Y et al. Prevalence of virulence and resistance to antibiotics in pathogenic enterococci isolated from mastitic cows. Journal of Veterinary Medical Science. 2016; 78 (11): 1663-1668.
29. Willis AT, Magdesian KG, Byrne BA, Edman JM. Enterococcus infections in foals. The Veterinary Journal 2019; 248: 42-47.
30. Song W, Park MJ, Kim HS, Meyer V, Hombach M. Comparison of Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing breakpoints for beta-lactams in Enterobacteriaceae producing extendedspectrum beta-lactamases and/or plasmid-mediated AmpC beta-lacta. Korean Journal of Clinical Microbiology 2011; 14: 24-9.
31. Poulou A, Grivakou E, Vrioni G, Koumaki V, Pittaras T et al. Modified CLSI extended-spectrum β-lactamase (ESBL) confirmatory test for phenotypic detection of ESBLs among Enterobacteriaceae producing various β-lactamases. Journal of Clinical Microbiology 2014; 52: 1483-1489.
32. Muslim SN, Mohammed Ali AN, Al-Kadmy IMS, Khazaal SS, Ibrahim SA et al. Screening, nutritional optimization and purification for phytase produced by Enterobacter aerogenes and its role in enhancement of hydrocarbons degradation and biofilm inhibition. Microbial Pathogenesis 2018; 115: 159-167.
33. Jamal M, Andleeb S, Jalil F, Imran M, Nawaz MA et al. Isolation, characterization and efficacy of phage MJ2 against biofilm forming multi-drug resistant Enterobacter cloacae. Folia Microbiologica 2019; 64: 101-111.
34. Sabir N, Ikram A, Zaman G, Satti L, Gardezi A et al. Bacterial biofilm-based catheter-associated urinary tract infections: causative pathogens and antibiotic resistance. American Journal of Infection Control 2017; 45 (10): 1101-1105.
35. Lupo A, Papp-Wallace KM, Sendi P, Bonomo RA, Endimiani A. Non-phenotypic tests to detect and characterize antibiotic resistance mechanisms in Enterobacteriaceae. Diagnostic Microbiology and Infectious Disease 2013; 77: 179-194.
36. Uludağ Altun H, Şener B. Biyofilm infeksiyonları ve antibiyotik direnci. Hacettepe Tıp Dergisi 2008; 39: 82-88 (in Turkish).
37. Slama Ben R, Kouidhi B, Zmantar T, Chaieb K, Bakhrouf A. Anti‐listerial and anti‐biofilm activities of potential probiotic Lactobacillus strains isolated from Tunisian traditional fermented food. Journal of Food Safety 2013; 33 (1): 8-16.
38. Deegan LH, Cotter PD, Colin H, Ross P. Bacteriocins: biological tools for bio-preservation and shelf-life extension. International Dairy Journal 2006; 16: 1058-1071.
39. Liao CC, Yousef AE, Chism GW, Richter ER. Inhibition of Staphylococcus aureus in buffer, culture media and foods by lacidin A, a bacteriocin produced by Lactobacillus acidophilus OSU133. Journal of Food Safety 1994; 14: 87-101.
40. Delves-Broughton J, Blackburn P, Evans RJ, Hugenholtz J. Applications of the bacteriocin, nisin. Antonie Van Leeuwenhoek 1996; 69: 193-202.
41. Santos VL, Nardi Drummond RM, Dias-Souza MV. Bacteriocins as antimicrobial and antibiofilm agents. Current Developments in Biotechnology and Bioengineering 2017; 403-436.
42. Cerning J. Exocellular polysaccharides produced by lactic acid bacteria. FEMS Microbiology Reviews 1990; 87: 113-130.
43. Tallon R, Bressollier P, Urdaci MC. Isolation and characterization of two exopolysaccharides produced by Lactobacillus plantarum EP56. Research in Microbiology 2003; 154 (10): 705-712. doi: 10.1016/j.resmic.2003.09.006
44. Looijesteijn PJ, Trapet L, de Vries E, Abee T, Hugenholtz J. Physiological function of exopolysaccharides produced by Lactococcus lactis. International Journal of Food Microbiology 2001; 64 (1-2): 71-80.
45. Karatuğ Taşkale N, Yüksel FN, Özdemir C, Gürcan C, Ugur S et al. Anti-biofilm potential effect of Lactococcus lactis CNM81 strain isolated from raw milk on Salmonella Typhimurium SL1344 biofilm. In: Proceedings of the 27th ECCMID Congress; Vienna, Austria; 2017.
46. Franz CM, Stiles ME, Schleifer KH, Holzappel WH. Enterococci in foods - a conundrum for food safety. International Journal of Food Microbiology 2003; 88 (2-3): 105-122.
47. Klein G. Taxonomy, ecology and antibiotic resistance of enterococci from food and gastro-intestinal tract. International Journal of Food Microbiology 2003; 88 (2-3): 123-131.
48. Franz CM, Holzapfel WH, Stiles ME. Enterococci at the crossroads of food safety. International Journal of Food Microbiology 1999; 47: 1-24.
49. Hugas M, Garria M, Aymerich MT. Functionality of enterococci in meat products. International Journal of Food Microbiology 2003; 88 (2-3): 223-233.
50. Saavedra L, Taranto MP, Sesma F, Valdez GF. Homemade traditional cheeses for the isolation of probiotic Enterococcus faecium strains. International Journal of Food Microbiology 2003; 88:241-245. doi: 10.1016/S0168-1605(03)00186-7
51. Soyuçok A, Ekiz T, Başyiğit Kılıç G. The properties of exopolysaccharides and their importance in food industry. Nevşehir Bilim ve Teknoloji Dergisi 2016; 332-344 (in Turkish with an abstract in English).
52. Böke H. Lactobacillus delbrueckii subsp. bulgaricus türüne ait susların bazı probiyotik özelliklerinin belirlenmesi ve tutuklamanın bu özellikler üzerine etkisi. MSc, Gazi University, Ankara, Turkey, 2005 (in Turkish).