Document Type : Original Article


1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Animal Science, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran

3 Department of Biology, Faculty of Sciences, Central Tehran Branch, Islamic Azad University, Tehran, Iran


The efficacy of probiotics as alternatives to antibiotics has been defined as one of the potential strategies to prevent Salmonella spp. infection in poultry. The purpose of this study was to isolate probiotic native Lactic acid bacteria (LAB) with high compatibility to intestinal tract and prevention of Salmonella typhimuriumfrom broiler chicken feces. Thirty-seven samples of chicken feces were collected from seven broiler chicken farms in Northern Iran. The isolates identification was carried out with morphological and biochemical tests. Agar diffusion methods were used to evaluate the antimicrobial activities against Escherichia coli and S. typhimurium. The primary probiotic characteristics such as resistance to acid and bile and adhesion to Caco-2 cells were studied. Indeed, the ability of LAB isolates to inhibit adhesion of S. typhimurium to Caco-2 cells was evaluated by exclusion, competition and displacement assays. Among 42 isolates, S08, S01 and S06 isolates which showed appropriate probiotics characteristics were selected. Isolates S08 and S01 showed to be able to adhere strongly and also S06 was adhered moderately. In the exclusion assay, the isolates S08, S01 and S06 significantly hampered adhesion of S. typhimurium cell, in the competition assay, the isolates S08, S01 showed significant level of competition activity against S. typhimurium adherence to Caco-2 cells and isolate S08 showed the greatest displacement activity. The 16S rDNA sequence revealed that S08, S01, and S06 isolates were 99.00% similar to Lactobacillus salivarius, Lactobacillus johnsonii, and Pediococcus acidilactici, respectively. The result of this study suggested that LAB isolated from broiler chicken feces could be a remarkable reservoir for identification of probiotic to inhibit the pathogenic bacteria growth.



    1. Williams BA, Verstegen MW, Tamminga S. Fermentation in the large intestine of single-stomached animals and its relationship to animal health. Nutr Res Rev 2001; 14(2): 207-228.
    2. Mishu B, Griffin PM, Tauxe RV, et al. Salmonella enteritidis gastroenteritis transmitted by intact chicken eggs. Ann Intern Med 1991; 115(3): 190-194.
    3. Sodagari HR, Mashak Z, Ghadimianazar A. Prevalence and antimicrobial resistance of Salmonella serotypes isolated from retail chicken meat and giblets in Iran. J Infect Dev Ctries 2015; 9(5): 463-469.
    4. Edens FW. An alternative for antibiotic use in poultry: Probiotics. Braz J Poultry Sci 2003; 5(2): 75-97.
    5. Hassanzadazar H, Ehsani A, Mardani K. Antibacterial activity of Enterococcus faecium derived from Koopeh cheese against Listeria monocytogenes in probiotic ultra-filtrated cheese. Vet Res Forum 2014; 5(3): 169-175.
    6. Taghi-Zadeh A, Nejati F. Screening of lactic acid bacteria isolated from Iranian sourdoughs for antifungal activity: Enterococcus faecium showed the most potent antifungal activity in bread. Appl Food Biotechnol 2017; 4(4): 219-227.
    7. Revolledo L, Ferreira AJ, Mead GC. Prospects in Salmonella control: Competitive exclusion, probiotics, and enhancement of avian intestinal immunity. J Appl Poult Res 2006; 15(2): 341-351.
    8. Rehman HU, Vahjen W, Awad WA. Indigenous bacteria and bacterial metabolic products in the gastrointestinal tract of broiler chickens. Arch Anim Nutr 2007; 61(5): 319-335.
    9. Shalaei M, Hosseini SM, Zergani E. Effect of different supplements on eggshell quality, some characteristics of gastrointestinal tract and performance of laying hens. Vet Res Forum 2014; 5(4): 277-286.
    10. Blajman J, Gaziano C, Zbrun MV, et al. In vitro and in vivo screening of native lactic acid bacteria toward their selection as a probiotic in broiler chickens. Res Vet Sci 2015; 101: 50-56.
    11. 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.
    12. Jacobsen CN, Nielsen VR, Hayford AE, et al. Screening of probiotic activities of forty-seven strains of Lactobacillus spp. by in vitro techniques and evaluation of the colonization ability of five selected strains in humans. Appl Environ Microbiol 1999; 65(11): 4949-4956.
    13. Gopal PK, Prasad J, Smart J, et al. In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their anta-gonistic activity against an enterotoxigenic Escherichia coli. Int J Food Microbial 2001; 67(3):207-216.
    14. Zhang YC, Zhang LW, Tuo YF, et al. Inhibition of Shigella sonnei adherence to HT-29 cells by lactobacilli from Chinese fermented food and preliminary characterization of S-layer protein involvement. Res Microbial 2010; 161(8): 667-672.
    15. Aquilina G, Bories G, Chesson A, et al. Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J 2012; 10(6): 2740. doi:10.2903/j.efsa.2012.2740.
    16. Carasi P, Díaz M, Racedo SM, et al. Safety characterization and antimicrobial properties of kefir-isolated Lactobacillus kefiri. Biomed Res Int 2014; 2014: 208974. doi: 10.1155/2014/208974.
    17. Touré R, Kheadr E, Lacroix C, et al. Production of antibacterial substances by bifidobacterial isolates from infant stool active against Listeria monocytogenes. J Appl Microbiol 2003; 95(5): 1058-1069.
    18. Chauviere G, Coconnier MH, Kerneis SO, et al. Adhesion of human Lactobacillus acidophilus strain LB to human enterocyte-like Caco-2 cells. Microbiology 1992; 138(8): 1689-1696.
    19. Candela M, Perna F, Carnevali P, et al. Interaction of probiotic Lactobacillus and Bifidobacterium strains with human intestinal epithelial cells: Adhesion properties, competition against enteropathogens and modulation of IL-8 production. Int J Food Microbiol 2008; 125(3): 286-292.
    20. Kim EY, Kim YH, Rhee MH, et al. Selection of Lactobacillus sp. PSC101 that produces active dietary enzymes such as amylase, lipase, phytase and protease in pigs. J Gen Appl Microbial 2007; 53(2): 111-117.
    21. Araújo WL, Angellis DA, Azevedo JL. Direct RAPD evaluation of bacteria without conventional DNA extraction. Braz Arch Biol Technol 2004; 47(3): 375-380. 
    22. Jannah SN, Dinoto A, Wiryawan KG, et al. Characteristics of lactic acid bacteria isolated from gastrointestinal tract of Cemani chicken and their potential use as probiotics. Media Peternakan 2014; 37(3): 182-189.
    23. Nemcova R. Criteria for selection of Lactobacillus for probiotic use. Vet Med (Praha) 1997; 42(1): 19-27.
    24. Shokryazdan P, Sieo CC, Kalavathy R, et al. Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. Biomed Res Int 2014; 2014: 927268. doi:10.1155/ 2014/927268.
    25. Charteris WP, Kelly PM, Morelli L, et al. Development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J Appl Microbial 1998; 84(5): 759-768.
    26. Corsetti A, Gobbetti M, Smacchi E. Antibacterial activity of sourdough lactic acid bacteria: Isolation of a bacteriocin-like inhibitory substance from Lactobacillus sanfrancisco C57. Food Microbial 1996; 13(6): 447-456.
    27. Jankowska A, Laubitz D, Antushevich H, et al. Competition of Lactobacillus paracasei with Salmonella enterica for adhesion to Caco-2 cells. Biomed Res Int BioMed J Biomed Biotechnol 2008; 357964. doi: 10.1155/2008/357964.
    28. Kizerwetter-Świda M, Binek M. Assessment of potentially probiotic properties of Lactobacillus strains isolated from chickens. Pol J Vet Sci 2016; 19(1): 15-20.
    29. Lim SM, Ahn DH. Factors affecting adhesion of lactic acid bacteria to Caco-2 cells and inhibitory effect on infection of Salmonella typhimurium. J Microbiol Biotechnol 2012; 22(12): 1731-1739.
    30. Gueimonde M, Jalonen L, He F, et al. Adhesion and competitive inhibition and displacement of human enteropathogens by selected lactobacilli. Food Res Int 2006; 39(4): 467-471.
    31. Lee YK, Puong KY, Ouwehand AC, et al. Displacement of bacterial pathogens from mucus and Caco-2 cell surface by lactobacilli. J Med Microbial 2003; 52(10): 925-930.
    32. Oh BC, Choi WC, Park S, et al. Biochemical properties and substrate specificities of alkaline and histidine acid phytases. Appl Microbiol Biotechnol 2004; 63(4): 362-372.