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Probiotic Potential and Antimicrobial Activity of Enterococcus faecium Isolated from Chicken Caecal and Fecal Samples

Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim


Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate, characterize and identify Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from chicken ceca and feces of thirty three clinically healthy chickens from a local farm. In vitro studies were performed to assess antibacterial activity of the isolated LAB (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified by biochemical carbohydrate fermentation patterns using an API 50 CHL kit and API ZYM kits and by sequenced 16S rDNA. An isolate belonging to E. faecium species exhibited inhibitory effect against S. enteritidis. This isolate producing a clear zone as large as 10.30 mm or greater and was able to grow in the coculture medium and at the same time, inhibited the growth S. enteritidis. In addition, E. faecium exhibited significant resistance under highly acidic conditions at pH 2.5 for 8 h and survived well in bile salt at 0.2% for 24 h and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. Enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: Antimicrobial activity, probiotic, acid tolerance, Enterococcus faecium

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[1] H. A. Abdel-Rahman., S. M. Shawky, H. Ouda, A. A. Nafeaa, and S. H. Orabi, "Effect of Two Probiotics and Bioflavonoids Supplementation to the Broilers Diet and Drinking Water on the Growth Performance and Hepatic Antioxidant Parameters". Global Veterinaria 10 (6): 734-741, 2013.
[2] R. Herich, and M. Levkut, "Lactic acid bacteria, probiotics and immune system". Veterinární medicína, 47, 169-180, 2002.
[3] P. Hlivak, J. Odraska, M. Ferencik, L. Ebringer, E. Jahnova, and A. Mikes, "One-year application of probiotic strain Enterococcus faecium M-74 decreases serum cholesterol levels". Bratisl. Lek. Listy 106, 67– 72, 2005.
[4] Y. Huang, and Y. Zheng, "The probiotic Lactobacillus acidophilus reduces cholesterol absorption through the down-regulation of Niemann–Pick C1-like 1 in Caco-2 cells". Br. J. Nutr. 9, 1–6, 2009.
[5] L. Pascual, F. Ruiz, W. Giordano, and I. L. Barberis, "Vaginal colonization and activity of the probiotic bacterium Lactobacillus fermentum L23 in a murine model of vaginal tract infection". Journal of Medical Microbiology 59, 360–364, 2010.
[6] O. Ashayerizadeh, B. Dastar, M. Shams Shargh, E. Rahmatnejad, and A. Ashayerizadeh, "Influence of prebiotic and two herbal additives on interior organs and hematological indices of broilers". Journal of Animal and Veterinary Advances. 8(9), p. 1851-1855, 2009.
[7] A. Bhardwaj, G. Kaur, H. Gupta, S. Vij, and R. K. Malik, "Interspecies diversity, safety and probiotic potential of bacteriocinogenic Enterococcus faecium isolated from dairy food and human faeces". World Journal of Microbiology and Biotechnology, 27, 591-602, 2011.
[8] J. A. Patterson, and K. M. Burkholder, "Application of prebiotics and probiotics in poultry production. Poultry Sci., 82: 627-631, 2003.
[9] A. Ahmadova, S. D. Todorov, Y. Choiset, H. Rabesona, T. M. Zadi, Kuliyev, A., B.D.G.M. Franco, J. M. Chobert, and T. Haertlé, "Evaluation of antimicrobial activity, probiotic properties and safety of wild strain Enterococcus faecium AQ71 isolated from Azerbaijani Motal cheese". Food Control 30, 631–64, 2013.
[10] B. B. Matijašic´, T. Obermajer, and I. Rogelj, "Quantification of Lactobacillus gasseri, Enterococcus faecium and Bifidobacterium infantis in a probiotic OTC drug by real-time PCR". Food Control 21, 419–425, 2010.
[11] L. Saavedra, M. P. Taranto, F. Sesma, and G. F. de Valdez, "Homemade traditional cheeses for the isolation of probiotic Enterococcus faecium strains". Int. J. Food Microbiol. 88, 241–245, 2003.
[12] J. Benyacoub, P. F. Perez, F. Rochat, K. Y. Saudan, G. Reuteler, N. Antille, M. Humen, G. L. De Antoni, C. Cavadini, S. Blum, and E. J. Schiffrin, "Enterococcus faecium SF68 enhances the immune response to Giardia intestinalisin mice". J. Nutr. 135: 1171–1176, 2005.
[13] D. G. V. Emmanuel, A. Jafari, K. A. Beauchemin, J. A. Leedle, and B. N. Ametaj, "Feeding live cultures of Enterococcus faecium and Saccharomyces cerevisiae induces an inflammatory response in feedlot steers". J. Anim. Sci. 85:233–239, 2007.
[14] U. Lodemann, K. Hubener, N. Jansen, and H. Martens, "Effects of Enterococcus faecium NCIMB 10415 as probiotic supplement on intestinal transport and barrier function of piglets". Arch. Anim. Nutr. 60:35–48, 2006.
[15] L. Scharek, J. Guth, K. Reiter, K. D. Weyrauch, D. Taras, P. Schwerk, P. Schierack, M. F. Schmidt, L. H. Wieler, and K. Tedin, "Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets". Vet. Immunol. Immunopathol. 105:151– 161, 2005.
[16] W. Theppangna, K. Otsuki, and T. Murase, "Inhibitory effects of Enterococcus strains obtained from a probiotic product on in vitro growth of Salmonella enteric serovar Enteritidis strain IFO3313". J. Food Prot. 69: 2258–2262, 2006.
[17] D. Taras, W. Vahjen, M.Macha, and O. Simon, "Performance, diarrhea incidence, and occurrence of Escherichia coli virulence genes during long-term administration of a probiotic Enterococcus faecium strain to sows and piglets". J. Anim. Sci. 84:608–617, 2006.
[18] M. Tokumaru, H. Konuma, M. Umesako, S.Konno, and K. Shinagawa, "Rates of detection of Salmonella and Campyobacterin meats in response to the sample size and the infection level of each species". Int. J. Food Microbiol. 13:41–46, 1991.
[19] M. A. Ehrmann, P. Kurzak, J. Bauer, and R. F. Vogel, "Characterization of lactobacilli towards their use as probiotic adjuncts in poultry". J. Appl. Microbiol. 92:966–975, 2002.
[20] L. M. Shin, R. J. McNally, S. M. Kosslyn, W. L. Thompson, S. L. Rauch, and N. M. Alpert, "Regional cerebral blood flow duringscriptdriven imagery in childhood sexual abuse-related posttraumatic stress disorder: A positron emission tomographic investigation". Am J Psychiatry 156:575–584, 1999.
[21] K. Perelmuter, M. Fraga, and P. Zunino, "In vitro activity of potential probiotic Lactobacillus murinus isolated from the dog". Journal of Applied Microbiology, 104, 1718-1725, 2008.
[22] Y. Huang, and M. C. Adams, "In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionibacteria". International Journal of Food Microbiology, 91, 253 -260, 2004.
[23] A. R. Madureira, M. S. Gião, M. E. Pintado, A. M. P. Gomes, C. Freitas, and F. X. Malcata, "Incorporation and survival of probiotic bacteria in whey cheese matrices". J. Food Sci. 70(3): M161-M165, 2005.
[24] M. Succi, P. Tremonte, A. Reale, E. Sorrentino, L. Grazia, and S. Pacifico, "Bile salt and acid tolerance of Lactobacillus rhamnosus strains isolated from Parmigiano Reggiano cheese". FEMS Microbiology Letters, 244, 129-137, 2005.
[25] S. Pieniz, R. Andreazza, T. Anghinoni, F. Camargo, and A. Brandelli, "Probiotic potential, antimicrobial and antioxidant activities of Enterococcus durans strain LAB18s. Food Control 37, 251-256, 2014.
[26] P. Sun, J. Wang, and Y. Jiang, "Effects of Enterococcus faecium (SF68) on immune function in mice". Food Chemistry, 123, 63-68, 2010.
[27] B. Hyronimus, C. L. Marrec, S. A. Hadj, and A. Deschamps, "Acid and bile tolerance of spore-forming lactic acid bacteria". International Journal of Food Microbiology, 61, 193-197, 2000.
[28] J. Gong, R., J. Forster, and H. Yu, "Diversity and phylogenetic analysis of bacteria in the mucosa of chicken ceca and comparison with bacteria in the cecal lumen". FEMS Microbiol Lett 208:1–7, 2002.
[29] J. B. Payne, J. A. Osborne, P. K. Jenkins, and B. W. Sheldon, "Modeling the growth and death kinetics of Salmonella in poultry litter as a function of pH and water activity". Poult. Sci., 86: 191-201, 2007.
[30] EFSA, The community summary report on trends and sources of zoonoses and zoonotic agents in the European Union in 2007. EFSA J 223:1–320, 2009.
[31] H. R. Taheri, H. Moravej, F. Tabandeh, M. Zaghari, and M. Shivazad, "Screening of lactic acid bacteria toward their selection as a source of chicken probiotic". Poult. Sci. 88:1586-1593, 2009.
[32] M. Chichlowski, J. Croom, B. W. McBride, G. Davis, L. Daniel, and M. Koci, "Direct-fed microbial and salinomycin modulate whole body and intestinal oxygen consumption and intestinal enterocytes cytokine production in the broiler chick". Poult. Sci., 86: 1100-1106, 2007.
[33] M. A. Riley, and J. E. Wertz, "Bacteriocins: evolution, ecology, and application". Annual review of microbiology, Vol.56, pp. 117-137, ISSN 0066-4227, 2002.
[34] P. Audisio, A.De Biase, G. Antonini, M. Belfiore, and M. Oliverio, "Morphological, molecular and ecological evidence of a new Euro- Anatolian species of the Meligethes coracinus complex (Coleoptera: Nitidulidae). Insect Systematics and Evolution, Volume 31, Issue 4, p. 361 – 385, 2000.
[35] R. Herich, T. Kokinčáková, A. Lauková, and M. Levkutová, "Effect of preventive application of Enterococcus faecium EF55 on intestinal mucosa during salmonellosis in Chickens". Czech J. Anim. Sci., 55, (1): 42–47, 2010.
[36] V. Strompfová, and A. Lauková, "In vitro study on bacteriocin production of enterococci associated with chickens". Anaerobe 13: 228- 237, 2007.
[37] M. Ashraf, and N. A. Akram, "Improving salinity tolerance of plants through conventional breeding and genetic engineering: an analytical comparison". Biotechnol. Adv., 27: 744-52, 2009.