Screening of Indigenous Strains of Lactic Acid Bacteria for Development of a Probiotic for Poultry

  • Karimi Torshizi, M.A. (Department of Poultry Science, College of Agriculture, Tarbiat Modares University Tehran) ;
  • Rahimi, Sh. (Department of Poultry Science, College of Agriculture, Tarbiat Modares University Tehran) ;
  • Mojgani, N. (Department of Biotechnology, Razi Vaccine and Serum Research Institute) ;
  • Esmaeilkhanian, S. (Department of Biotechnology, Animal Science Research Center) ;
  • Grimes, J.L. (Department of Poultry Science, North Carolina State University)
  • Received : 2008.01.16
  • Accepted : 2008.05.06
  • Published : 2008.10.01


In an attempt to develop a probiotic formulation for poultry feed, a number of lactic acid bacteria (LAB) were isolated from chicken intestinal specimens and a series of in vitro experiments were performed to evaluate their efficacy as a potential probiotic candidate. A total of 650 LAB strains were isolated and screened for their antagonistic potential against each other. Among all the isolates only three isolates (TMU121, 094 and 457) demonstrated a wide spectrum of inhibition and were thus selected for detailed investigations. All three selected isolates were able to inhibit the growth of E. coli and Salmonella species, although to variable extent. The nature of the inhibitory substance produced by the isolates TMU121 and 094 appeared to be associated with bacteriocin, as their activity was completely lost after treatment with proteolytic enzymes, while pH neutralization and catalase enzyme had no effect on the residual activity. In contrast, isolate TMU457 was able to resist the effect of proteolytic enzymes while pH neutralization completely destroyed its activity. Attempts were made to study the acid, bile tolerance and cell surface hydrophobicity of these isolates. TMU121 showed high bile salt tolerance (0.3%) and high cell surface hydrophobicity compared to the other two strains studied, while TMU094 appeared the most pH resistant strain. Based on these results, the three selected LAB isolates were considered as potential ingredients for a chicken probiotic feed formulation and were identified to species level based on their carbohydrate fermentation pattern by using API 50CH test kits. The three strains were identified as Lactobacillus fermentum TMU121, Lactobacillus rhamnosus TMU094, and Pediococcus pentosaceous TMU457.


Lactic Acid Bacteria;Probiotics;In vitro;Broiler


  1. Gilliland, S. E. and D. K. Walker. 1990. Factors to consider when selecting a culture of Lactobacillus acidophilus used as a dietary adjunct to produce a hypercholesterolemic effect in humans. J. Dairy Sci. 73:905-909.
  2. Gilliland, S. E., C. R. Nelson and C. Maxwell. 1985. Assimilation of cholesterol by Lactobacillus acidophilus. Appl. Environ. Microbiol. 49:377-381.
  3. Gilliland, S. E., T. E. Staley and L. J. Bush. 1984. Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct. J. Dairy Sci. 67:3045-3051.
  4. Gupta, P. K., B. K. Mital and S. K. Garg. 1996. Characterization of Lactobacillus acidophilus strains for use as dietary adjunct. Int. J. Food Microbiol. 29:105-109.
  5. Gusils, C., A. Perez Chaia, S. Gonzalez and G. Oliver. 1999. Lactobacilli isolated from chicken intestines: Potential use as probiotics. J. Food Protect. 62:252-256.
  6. Garriga, M., M. Pascal, J. M. Montfort and M. Hugas. 1998. Selection of lactobacilli for chicken probiotic adjuncts. J. Appl. Microbiol. 84:125-132.
  7. Ghadban, G. S. 2002. Probiotics in broiler production- a review. Arch. Geflugelk. 22:49-58.
  8. Gibson, G. R., P. B. Otaway and A. R. Robert. 2000. Prebiotics: new developments in functional foods. Chandos Publishing (Oxford) Limited, London, UK.
  9. Gilliland, S. E. and M. L. Speck. 1977. Deconjugation of bile acids by intestinal lactobacilli. Appl. Environ. Microbiol. 33:15-18.
  10. Corzo, G. and S. E. Gilliland. 1999. Measurement of bile salt hydrolase activity from Lactobacillus acidophilus based on disappearance of conjugated of bile salts. J. Dairy Sci. 82:466- 471.
  11. Delves-Broughton, J. 1990. Nisin and its uses as a food preservative. Food. Technol. 44:100-112.
  12. Erkkila, S. and E. Petaja. 2000. Screening of the commercial meat starter cultures at low pH and in the presence of bile salts for potential use of probiotic use. Meat Sci. 55:297-300.
  13. Fuller, R. 1989. A review: Probiotics in man and animals. J. Appl. Bacteriol. 66:365-378.
  14. Begley, M., C. G. M. Gahan and C. Hill. 2005. The interaction between bacteria and bile. FEMS Microbiol. Rev. 29:625-651.
  15. Buchanan, R. E. and N. E. Gibbons. 1994. Bergey's manual of determinative bacteriology, 9th ed. Williams & Wilkins, Baltimore.
  16. Tortuero, F. 1973. Influence of the implantation of Lactobacillus acidophilus in chicks on the growth, feed conversion, malabsorption of fats syndrome and intestinal flora. Poult. Sci. 52:197-203.
  17. Suskovic, J., B. Kos, J. Goreta and S. Matosic. 2001. Role of lactic acid bacteria and bifidobacteria in synbiotic effect. Food Technol. Biotechnol. 39:227-235.
  18. Ten Brink, B., M. Minekus, J. M. B. M. van der Vossen, R. J. Leer and J. H. J. Huis in't Veld. 1994. Antimicrobial activity of lactobacilli: preliminary characterization and optimization of production of acidocin B, a novel bacteriocin produced by Lactobacillus acidophilus M46. J. Appl. Bacteriol. 77:140-148.
  19. Toit, M. D., C. M. A. P. Franz, L. M. T. Dicks, U. Schillinger, P. Haberer, B. Warlies, F. Ahrens and W. H. Holzapfel. 1998. Characterization and selection of probiotic lactobacilli for a preliminary minipig feeding trial and their effect on serum cholesterol levels, faeces pH and faeces moisture content. Int. J. Food Microbiol. 40:93-104.
  20. Schillinger, U. and F. K. Lucke. 1989. Antibacterial activity of Lactobacillus sake isolated from meat. App. Environ. Microbiol. 55:1901-1906.
  21. Sjovall, J. 1959. Dietary glycine and taurine on bile acid conjugation in man; bile acids and steroids. Proc. Soc. Exp. Biol. Med. 100:676-678.
  22. Steel, R. G. D. and J. H. Torrie. 1980. Principles and Procedures of Statistics: A Biometrical Approach. 2nd edn. McGraw-Hill Book Company, New York.
  23. Nurmi, E. and M. Rantala. 1973. New aspects of Salmonella infection in broiler production. Nature 241:210-211.
  24. Patterson, J. A. and K. M. Burkholder. 2003. Application of probiotics and probiotics in poultry production. Poult. Sci. 82: 627-631.
  25. Prasad, J., H. Gill, J. Smart and P. K. Gopal. 1999. Selection and characterization of lactobacillus and bifidobacterium strains for use as probiotics. Int. Dairy J. 12:993-1002.
  26. Reid, G. and R. Friendship. 2002. Alternatives to antibiotic use: Probiotics for the gut. Anim. Biotechnol. 13:92-97.
  27. SAS Institute Inc. 1990. SAS/STAT User's Guide: Version 6. 4th edn. SAS institute Inc., Carolina.
  28. Lewus, C. B., A. Kaiser and T. J. Montville. 1991. Inhibition of food-borne bacterial pathogens by bacteriocins from lactic acid bacteria isolated from meat. Appl. Environ. Microbiol. 57:1683-1688.
  29. Marounek, M. and V. Rada. 1995. Susceptibility of poultry Lactobacilli to ionophore antibiotics. J.Vet. Med. 42:193-196.
  30. Mayra-Makinen, A., M. Manninen and M. Gyllenberg. 1983. The adherence of lactic acid bacteria to the columnar epithelium cells of pigs and calves. J. Appl. Bacteriol. 55:241-245.
  31. McKay, L. L. and K. A. Boldwin. 1990. Applications for biotechnology: present and future improvements in lactic acid bacteria. FEMS Microbiol. Rev. 87:3-14.
  32. Mulder, R. W. A. W., R. Havenaar and J. H. J. Huis in't Veld. 1997. Intervention strategies: the use of probiotics and competitive exclusion microfloras against contamination with pathogens in pigs and poultry. In: Probiotics 2. Applications and practical aspects (Ed. R. Fuller). Chapman and Hall, London, UK. pp. 187-205.
  33. Nemcova, R., A. Laukova, S. Gancarcikova and R. Kastel. 1997. In vitro studies of porcine lactobacilli for possible probiotic use. Berl. Munch. Tierarztl. Wschr. 110:413-417.
  34. Juven, B. J., R. J. Meinersmann and N. J. Stern. 1991. Antagonistic effects of Lactobacilli and Pediococci to control intestinal colonization by human enteropathogens in live poultry. J. Appl. Bacteriol. 70:95-103.
  35. Juven, B. J., F. Schved and P. Linder. 1992. Antagonistic compounds produced by a chicken intestinal strain of Lactobacillus acidophilus. J. Food Protect. 55:157-161.
  36. Kim, G. B. and B. H. Lee. 2005. Biochemical and molecular insights into bile salt hydrolase in the gastrointestinal microflora - A review. Asian-Aust. J. Anim. Sci. 18:1505-1515.
  37. Kim, H. J., H. S. Shin, W. K. Ha, I. H. Yang and S. W. Lee. 2006. Characterization of lactic bacterial strains isolated from raw milk. Asian-Aust. J. Anim. Sci. 19:131-136.
  38. Lankaputhra, W. E. and N. P. Shah. 1995. Survival of Lactobacillus acidophilus and Bifidobacterium spp. in the presence of acid and bile salts. Cult. Dairy Prod. J. 30:2-7.
  39. Gusils, C., M. Bujazha and S. Gonzalez. 2002. Preliminary studies to design a probiotic for use in swine feed. Interciencia. 27:409-413.
  40. Hood, S. K. and A. Zottola. 1988. Effect of low pH on the ability of Lactobacillus acidophilus to survive and adhere to human intestinal cells. J. Food Sci. 53:1514-1516.
  41. Jin, L. Z., W. Y. Ho, M. A. Abdullah and S. Jalaludin. 1998a. Growth performance, intestinal microbial populations, and serum cholesterol of broilers fed diets containing lactobacillus cultures. Poult. Sci. 77:1259-1265.
  42. Jin, L. Z., W. Y. Ho, M. A. Abdullah, M. A. Ali and S. Jalaludin. 1998b. Effects of adherent lactobacillus cultures on growth, weight of organs and intestinal microflora and volatile fatty acids in broilers. Anim. Feed Sci. Technol. 70:197-209.
  43. Jin, L. Z., W. Y. Ho, M. A. Abdullah, M. A. Ali and S. Jalaludin. 1996. Antagonistic effects of intestinal lactobacillus isolates on pathogens of chicken. Lett. Appl. Microbiol. 23:67-71.
  44. Joerger, M. C. and T. R. Klaenhammer. 1986. Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481. J. Bacteriol. 167:439-446.
  45. Wadström, T., K. Andersson, M. Sydow, L. Axelsson, S. Lindgren and B. Gullmar. 1987. Surface properties of lactobacilli isolated from the small intestine of pigs. J. Appl. Bacteriol. 62: 513-520.
  46. Yu, B., J. R. Liu, M. Y. Chiou, Y. R. Hsu and P. W. S. Chiou. 2007. The effects of probiotic Lactobacillus reuteri Pg4 strain on intestinal characteristics and performance in broilers. Asian- Aust. J. Anim. Sci. 20:1243-1251.
  47. Treagan, L. and L. Pulliam. 1982. Medical microbiology laboratory procedures. W.B. Saunders Company, Philadelphia. pp. 233-243.
  48. Upreti, G. C. and R. D. Hinsdill. 1975. Production and mode of action of Lactocin 27: bacteriocin from a homofermentative lactobacillus. Antimicrob. Agents Ch. 7:139-145.
  49. Usman, H. A. 1999. Bile tolerance, taurocholate deconjugation, and binding of cholesterol by Lactobacillus gasseri strains. J. Dairy Sci. 82:243-248.
  50. Collins, J. K., G. Thornton and G. O. Sullivan. 1998. Selection of probiotic strains for human applications. Int. Dairy J. 8:487- 490.
  51. Dharmawan, J., I. S. Surono and L. Y. Kun. 2006. Adhesion properties of indigenous Dadih lactic acid bacteria on human intestinal mucosal surface. Asian-Aust. J. Anim. Sci. 19:751- 755.
  52. Huis in't Veld, J. H. J. and C. Shortt. 1996. Selection criteria for probiotic micro-organisms. In: Gut flora and health- past, present and future (Ed. A. R. Leeds and I. R. Rowland). International Congress and Symposium Series No. 219, Royal Society of Medicine Press Limited. pp. 27-36.

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