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Asian Journal of Dairy and Food Research, volume 35 issue 1 (march 2016) : 50-57

Selection and characterization of potential probiotic Lactobacilli spp isolated from chicken feces may be used as a potent antibacterial agent

Nazila Aazami1, Enayatollah Kalantar2, Helen Poormazaheri2, Nassimeh Setayesh vali pour3, Gholamreza Salehi Jouzan*
1<p>Microbial Biotechnology &amp; Biosafety&nbsp;Department,&nbsp;Agricultural Biotechnology, Institute of Iran, Karaj, Iran.</p>
Cite article:- Aazami1 Nazila, Kalantar2 Enayatollah, Poormazaheri2 Helen, pour3 vali Setayesh Nassimeh, Jouzan* Salehi Gholamreza (2016). Selection and characterization of potential probiotic Lactobacilli spp isolated from chicken feces may be used as a potent antibacterial agent . Asian Journal of Dairy and Food Research. 35(1): 50-57. doi: 10.18805/ajdfr.v35i1.9252.

ABSTRACT

Lactobacillus strains commonly used as probiotics which possess some properties such as resistance to gastric acidity and bile salts, production of antimicrobial compounds, the ability to modulate immune responses, and adhesion to gut tissues. This study aimed to isolate, identify, production and characterization of the probiotic strains. Total of 10 digestive tracts of chickens fed without antibiotics for presence of Lactobacillus spp was evaluated. All isolates were grown with different conditions like effect of temperature, pH, pepsine, bile, sodium chloride tolerance, aggregation, adhesion and antibiotic resistance test tested and then antibacterial activity was determined.In this study, 34 lactic acid bacteria were investigated as putative probiotic candidates. Based on the molecular identification using 16S rDNA sequencing the isolates belonged to  Lactobacillus species. The strains showed potent antibacterial activity. The results on acid tolerance showed that all the strains tested survived at pH 2.0 to 5.5. L. agilis strain M18, L. salivarius strain M10, L.vaginalis strains M8 and M19, and L. crispatus strains Or2 and M10 showed the lowest sensitivity to pepsine. All the studied Lactobacillus strains were able to grow at both 15 and 45°C . Five Lactobacillus strains adhered well to the Caco-2 cells. The result of antibiotic resistance test of 34 lactic acid bacteria showed that most of them were resistant or intermediate. Based on 34 Lactobacillus strains,  L. vaginalis strain M8 and  L. reuteri strain M6 were selected for the further in vivo assays and possible eventual use as prophylactic and therapeutic agent.

REFERENCES


  1. Abdulla, A., Abed, T.A., and Saeed, A.M. (2014). Adhesion auto aggregation and hydrophobicity of six Lactobacillus strains. British Microbiology Research. 4: 381-391.

  2. Arihara, K., Ota, H., Itoh, M., Kondo, Y., Sameshima, T., Yamanaka, H., Akimoto, M., Kanai, S., and Miki, T. (1998). Lactobacillus acidophilus group lactic acid bacteria applied to meat fermentation. J. Food Sci.63: 544-547. 

  3. Azami, N., Salehi Jouzani, G., Khodaei, Z., Meimandipour, A., Safari, M., Goudarzvand, M. (2014). Isolation, identification and characterization of some potentially probiotic lactobacillus strains from Iranian (Isfahan) native chickens. J. Gen. Appl. Microbiol. 60: 215-221.

  4. Baltimore, MD. (2009). Bergey’s Manual of Systematic Bacteriology, Williams and Wilkins. 9th ed. 2: 1063-1065.

  5. Bauer, AW., Kirby, WM., Sherris, JC., Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 45:493-6. 

  6. Bernet Camard, MF., Liévin, V., Brassart, D., Neeser, JR., Servin, AL., Hudault, S. (1997). The human Lactobacillus acidophilus strain LA1 secretes a nonbacteriocin antibacterial subtance(s) active in vitro and in vivo. Applied and EnvironmentalMicrobiology. 63: 2747–53.

  7. Collado, M.C., and Sanz, Y. (2006). Method for direct selection of potentially probiotic Bifidobacterium strains from human feces based on their acid-adaptation ability. Journal of Microbiological Methods.66: 560–563.

  8. Da Cunha, L. R., Fortes Ferreira L. L., Durmaz, E., Jun Goh, Y., Sanozky-Dawes, R. B., and Todd, T.R. (2102). Characterization of Lactobacillus gasseri isolates from a breast-fed infant. Gut Microbes. 3: 15–24.

  9. Danielsen, M. and Wind, AA. (2003). Susceptibility of Lactobacillus spp. to antimicrobial agents. Int J Food Microbiol. 82:1-11; 

  10. Dunne, C., OMahony, L., Murphy, L., Thornton, G., Morrissey, D., O’Halloran, S., Feeney ,M., Flynn, S., Fitzgerald, G., Daly, C., Kiely, B., O’Sullivan, G.C., Shanahan, F., and Collins, J.K. (2001). In vitro selection criteria for probiotic bacteria of human origin: Correlation in vivo findings. Am. J. Clin. Nutr.73: 386-392.

  11. Erkkila, S. and Petaja, E. (2000). Screening of commercial meat starter cultures at low pH in the presence of bile salts for potential probiotic use. J. Meat Sci.55:297-300. 

  12. Garriga, M., Pascual, M., Monfort, J.M., and Hugas, M. (1998). Selection of lactobacilli for chicken probiotic adjuncts. J. Appl. Microbiol.84: 125–132.

  13. Granato, D., Bergonzelli, E., Pridmore, R. D., Marvin, L., Rouvet, M., and Corthe´sy-Theulaz, E. 2004.Cell Surface-    Associated Elongation Factor Tu Mediates the Attachment of Lactobacillus johnsonii NCC533 (La1) to Human Intestinal Cells and Mucins. Infection and immunity. 72: 2160.

  14. Hsieh, PS., An, Y., Tsai, YC., Chen, YC., Chuang, CJ., Zeng, CT., Wang, CT., King, VA. 2013. Potential of probiotic strains to modulate the inflammatory responses of epithelial and immune cells in vitro. New Microbiologica. 36:167-179.

  15. Hummel, A.S., Hertel, C., Holzapfel, H.W., and Franz, C.M.A.P. (2007). Antibiotic Resistances of Starter and Probiotic Strains of Lactic Acid Bacteria. Applied And Environmental Microbiology.73: 730–739. 

  16. Jacobsen, C.N., Nielsen, V.R., Hayford, A.E., Moller, P.L., Michaelsen, K.F., Paerregaard, A., Sandstrom, B., Tvede, M., Jakobsen, M. (1999). 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.

  17. 65: 4949–4956.

  18. Lawalata, H. J., Sembiring, L., Rahayu, E.S. (2011). Molecular identification of lactic acid bacteria producing antimicrobial agents from Bakasang, an Indonesian traditional fermented fish product. Indonesian Journal of Biotechnology.

  19. 16: 93-99.

  20. Lawalata, H.J., Sembiring, L., and Rahayu, E.S. (2011). Molecular identification of lactic acid bacteria producing antimicrobial agents from Bakasang, an Indonesian traditional fermented fish product. Indonesian Journal of Biotechnology. 16: 93-99.

  21. Liasi, S.A., Azmi, T.I., Hassan, M.D., Shuhaimi, M., Rosfarizan, M., and Ariff, A.B. (2009). Antimicrobial activity and antibiotic sensitivity of three isolates of lactic acid bacteria from fermented fish product Budu. Malaysian Journal of Microbiology. 5: 33-37

  22. Michlmayr, H., Schümann, C., BarreiraBraz, N.M., Kulbe, K.D., Del Hierro, A.M. (2010). Isolation and basic characterization of a â-glucosidase from a strain of Lactobacillus brevis isolated from a malolactic starter culture. Journal of Applied Microbiology. 108: 550–559. 

  23. Miyamoto, T., Horie, T., Fujiwara, T., Fukata, T., Sasai, K., and Baba, E. (2000). Lactobacillus Flora in the Cloaca and Vagina of Hens and Its Inhibitory Activity Against Salmonella enteritidis In vitro. Poult. Sci.79:7-11.

  24. Perea Velez, M., Hermans, K., Verhoeven, T.L.A., Lebeer, S.E., Vanderleyden, J., De, S.C.J. (2007). Identification and characterization of starter lactic acid bacteria and probiotics from Columbian dairy products. Journal of Applied Microbiology. 103: 666-674.

  25. Reniero, R., Cocconcelli, P., Bottazzi, V., and Morelli, L. (1992). High frequency of conjugation in Lactobacillus mediated by an aggregation-promoting factor. Microbiology.138: 763- 768.

  26. Savino, F., Cordisco, L., Tarasco Locatelli, E., Di Gioia, D., Oggero, R., and Matteuzzi, M. (2011). Antagonistic effect of Lactobacillus strains against gas-producing coliforms isolated from colicky infants. BMC Microbiology. 11:157- 164.

  27. Thomas, P. (2007). Isolation and identification of five Alcohol-defying Bacillus spp. covertly associated with in vitro culture of seedless watermelon. Curr. Sci. 92: 983-986.

  28. Van Coillie, E., Goris, J., Cleenwerck, I., Grijspeerdt, K., Botteldoorn, N., and Van Immerseel, F. (2007). Identification of Lactobacilli isolated from the cloaca and vagina of laying hens and characterization for potential use as probiotics to control Salmonella enteritidis. J. Appl.Microbiol.102:1095- 1106.

  29. Yamazaki, M., Ohtsu H,Yakabe, Y., Kishima, M., and Abe, H. (2012). In vitro screening of lactobacilli isolated from chicken excreta to control Salmonella enteritidis and typhimurium. British Poultry Science.53:183-189

     
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