Inhibitory effect of native enterococci isolates on some of the foodborne bacterial pathogens

Subject Areas : Food Science and Technology

S. Tafkiki ¹ , S. Hanifian ²

1 - M.Sc Graduate of Food Science and Technology, Tabriz Branch, Islamic Azad University, Tabriz, Iran
2 - Associate Professor of Department of Food Science and Technology, Tabriz Branch, Islamic Azad University, Tabriz, Iran

Received: 2018-04-10 Accepted : 2018-05-05 Published : 2019-03-21

Keywords: Enterococcus, raw milk, Traditional products, Antagonistic effect, Dual layer method,

Abstract :

Enterococci are among lactic acid bacteria that are homogeneous and are commonly found in raw milk and its products. The purpose of this study was to isolate Enterococcus species from raw milk and traditional milk products in Tabriz region and to study their inhibitory effect on some of pathogenic bacteria. For this, 105 specimens including 15 samples of each of raw milk, yogurt, cheese, cream, butter, dough, and whey were tested. After isolation and differential identification of the species, 24 isolates were selected based on the sample type and the variety of Enterococcus species. Due to the pathogenic potential of E. faecalis and E. fascism species, they were evaluated for the presence of Esp < /em> and Asa1 genes, respectively. The inhibitory effect of Enterococcus isolates was tested on nine important foodborne pathogens using overlay method. According to the results, nine species of E. fascism, E. faecalis, E. gallinarum, E. avium, E. Benedetti, E. caseliflavus, E. hirae, E. saccharoliticus and E. raffinosus were isolated. All isolates had an inhibitory effect on indicator organisms; however, the inhibitory effect was found different among various species and various strains. E. faecalis and E. fascism species had the most antibacterial effects. The molecular evaluation showed that out of 5 isolates of E. faecalis, 2 isolates contained Esp < /em> gene. In the case of E. faecalis, none of the isolates harbored Asa1 gene. If further experiments are carried out in relation to the proven safety of enterococci, their inhibitory effect on food pathogenic bacteria can be used.

References:

●Aguilar, C., Vanergas, C. and Klotz, B. (2011). Antagonistic effect of Lactobacillus strains against Escherichia coli and Listeria monocytogenes in milk. Journal of Dairy Research, 78(2): 136–143.
●Alvarado, García Almendárez, B.E. , Martin, S.E. and Regalado, C. (2006). Food-associated lactic acid bacteria with antimicrobial potential from traditional Mexican foods. Revists Latinoamericano De Microbiologia, 48(3–4): 260–268.
●Arribas, P., Seseña, S., Poveda, J.M., Chicón, R., Cabezas, L. and Palop, L. (2011). Enterococcus populations in artisanal Manchego cheese: Biodiversity and safety aspects. Food Microbiology, 28: 891–899.
●Balouiri, M., Sadiki, M. and Ibnsouda, S.K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2): 71–79.
●Carasi, P., Jacquot, C., Romanin, D.E., Elie, A.M., De Antoni, G.L., Urdaci, M.C. et al. (2014). Safety and potential beneficial properties of Enterococcus strains isolated from kefir. International Dairy Journal, 39(1): 193–200.

●Cleveland, J., Montville, T.J., Nes, I.F. and Chikindas, M.L.  (2001). Bacteriocins: safe, natural antimicrobials for food preservation. International Journal of Food Microbiology, 71(1): 1–20.

●Dever, L.L. and Handwerger, S. (1996). Persistence of Vancomycin-resistance Enterococcus faecium gastrointestinal tract colonization antibiotic-treated mice. Microbe Drug Resistant, 2: 415–421.

●Ferguson, D.M., Talavera, G.N., Hernández, L.A.R., Weisberg, S.B., Ambrose, R.F. and Jay, J.A. (2016). Virulence Genes among Enterococcus faecalis and Enterococcus faecium isolated from coastal beaches and human and nonhuman sources in southern California and Puerto Rico. Journal of Pathogens, 3437214.
●Fisher, K. and Phillips, C. (2009). The ecology, epidemiology and virulence of Enterococcus. Microbiology, 155, 1749–1757.

●Fortina, M.G., Ricci, G., Borgo, F., Manachini, P.L., Arends, K., Schiwon, K., Abajy, M.Y., Grohmann, E. (2008). A survey on biotechnological potential and safety of the novel Enterococcus species of dairy origin, E. italicus. International Journal of Food Microbiology, 123: 204–211.

●Fracalanzza, S.A.P., Scheidegger, E.M.D., Santos, P.F.D., Leite, P.C. and Teixeira, L.M. (2007). Antimicrobial resistance profiles of Enterococci isolated from poultry meat and pasteurized milk in Rio de Janeiro, Brazil.  Mem Inst Oswaldo Crus, Rio de Janeiro, 102(7): 853–859.

●Franz, C.M.A.P., Stiles, M.E., Schleifer, K.H and Holzapfel, W.H. (2003). Enterococci in foods-a conundrum for food safety. International Journal of Food Microbiology, 83: 105–122.

●Giraffa, G. (2003). Functionality of enterococci in dairy products. International Journal of Food Microbiology, 88: 215–222.

●Gomes, B.C., Esteres, C.T., Palozzo, I.C.V., Darini, A.L.C., Felis, G.E., Sechi, L.A. et al. (2008). Prevalence and characterization of Enterococcus spp. isolated from Brazilian foods. Journal of Food Microbiology, 25: 668–675.

●Granato, D., Branco, G.F., Gomes Cruz, A., Faria, J.A.F., Shah, N.P. (2005). Probiotic dairy products as functional foods. Comprehensive Reviews in Food Science and Food Safety, 9(5): 455–470.

●Jamet, E., Akary, E., Poission, M.A. Cham, J.F., Bertrand, X. and Serror, P. (2012). Prevalence and characterization of antibiotic-resistant Enterococcus feacalis in French cheese. Food Microbiology, 31, 191–198.

●Heidari, H., Hasanpour, S., Ebrahim-Saraie, H.S. and Motamedifar, M. (2017). High incidence of virulence factors among clinical Enterococcus faecalis isolates in southwestern Iran. Infection & Chemotherapy, 49(1): 51–56.

●Hockett K.L. and Baltrus, D.A. (2017). Use of the soft-agar overlay technique to screen for bacterially produced inhibitory compounds. Journal of Visualized Experiments, (119): 55064.

●Lei, M., Dai, X. and Liu, M. (2015). Biological characteristics and safety examination of five Enterococcal strains from probiotic products. Journal of Food Safety, 35(3): 324–335.

●Manero, A. and Blanch, A.R. (1999). Identification of Enterococcus spp. with a biochemical key. Applied and Environmental Microbiology, 65(10): 4425–4430.

●Mannu, L., Paba, A., Daga, E., Comunian, R., Zanetti, S., Dupre, I. et al. (2013). Comparison of the incidence of virulence determinants and antibiotic resistance between Enterococcus faecium strains of dairy, animal and clinical origin. International Journal of Food Microbiology, 88: 291–304.

●Morandi, S., Brasca, M., Andrighetto, C., Lombardi, A. and Lodi, R. (2006). Technological and molecular characterization of enterococci isolated from north-west Italian dairy products. International Dairy Journal 16: 867–875.

●Moreno, M.R.F., Sarantinopoulos, P., Tsakalidou, E and De Vuyst, L. (2006). The role and application of Enterococci in food and health. International Journal of Food Microbiology, 106(1): 1–24.

●Nam, H.M., Kim, J.M., Moon, J.S., Kang, H.M., Jang, K.C., Joo, Y.S. et al. (2009). Antimicrobial  resistance  of  Enterococci  isolated  from mastitis  bovine  milk  samples  in  Korea. Zoonoses and Public Health, National Veterinary Research and Quarantine Service, Anyang City, Gyeonggi-d Korea, 48, 430–436.

●Polak-Berecka, M., Waśko, A. and  Koston, D. (2009). Comparison of different methods for detection of antimicrobial activity of probiotic strains of Lactobacillus rhamnosus against some food spoilage microorganisms. Annales UMCS, Biologia, 64: 15–24.

●Rashid, S. Hasan Shahiyan, M., Saeedi Asl, M.R. and Estiri, H. (2015). Screening, identification and evaluation of probiotic potential of Enterococci in traditional dairy products of Sabzevar. Journal of Sabzevar University of Medical Sciences, 21 (1): 7–16.

●Rasouli Pirouzian, H., Hesari, J., Farajnia, S., Moghaddam, M. and Ghiassifar, Sh. (2009). Isolation and identification of dominant strains of enterococci in traditional lighvan cheese. Journal of Food Research, 19(1): 13–24. [In Persian]

●Sánchez Valenzuela, A., ben Omar, N., Abriouel, H., Lucas Lopez, R., Veljovic, K., Martinez Canamero, M. et al. (2009). Virulence factors, antibiotic resistance, and bacteriocins in enterococci from artisan foods of animal origin. Food Control, 20, 381–385.

●Serio, A., Chaves-López, C., Paparella, A. and Suzzi, G. (2010). Evaluation of metabolic activities of enterococci isolated from Pecorino Abruzzese cheese. International Dairy Journal 20: 459–464.

●Uraipan, S. and Hongpattarakere, T. (2015). Antagonistic characteristics against food-borne pathogenic bacteria of lactic acid bacteria and bifidobacteria isolated from feces of healthy Thai infants. Jundishapur Journal of Microbiology, 8(6): 18264.

●Vankerckhoven, V., Van Autgaerden, T, Vael, C., Lammens, C., Chapelle, S., Rossi, R. et al. (2004). Development of a multiplex PCR for the detection of asa1, gelE, cylA, esp, and hyl genes in enterococci and survey for virulence determinants among European hospital isolates of Enterococcus faecium. Journal of Clinical Microbiology, 42(10): 4473–4479.

●Yoon, M.Y., Kim, Y.J. and Hwang, H.J. (2008). Properties and safety aspects of Enterococcus faecium strains isolated from Chungkukjang, a fermented soy product. LWT-Food Science and Technology, 41(5): 925–933.