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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 5 (may 2019) : 655-660,   Doi: 10.18805/ijar.B-3547

In vitro selection of potential immunomodulatory multispecies lactic acid bacteria in broilers

K
K. Divya Manjari
G
G. Divya
T
T.S. Saravanan
M
M. Parthiban
1Department of Animal Biotechnology, Madras Veterinary College, Faculty of Basic Sciences, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 007, Tamil Nadu, India.
Cite article:- Manjari Divya K., Divya G., Saravanan T.S., Parthiban M. (2018). In vitro selection of potential immunomodulatory multispecies lactic acid bacteria in broilers. Indian Journal of Animal Research. 53(5): 655-660. doi: 10.18805/ijar.B-3547.

ABSTRACT

Modulation of host immunity is one of the proposed benefits of the consumption of probiotics.  In this study a total of 26 lactic acid bacteria belonging to eight different species were isolated from chicken gut. These twenty six probiotic organisms were characterized by Gram’s staining, biochemical methods using 21 sugars, pH tolerance and bile salt tolerance test.  These probiotic organisms were identified using PCR amplification of 16S-23S spacer region and sequencing analysis. Based on the sequencing data, 6 different species were found to exist namely Enterococcus faecalis; Enterococcus faecium, Enterococcus hirae, Weissella cibaria, Lactobacillus salivarius, Pediococcus acidilactici. To select the immunomodulatory probiotic bacteria, the scoring index was established based on different chicken cytokine expression and B and T cell receptor expression studies. According to scoring index, Pediococcus acidilactici, Lactobacillus salivarius and Weissellacibaria showed highest levels of fold increase in the expression of Th1 cytokines viz. IL2, IL 12p40, IL1b and TNF a. Enterococcus faecalis and Enterococcus faecium showed highest levels of fold increase in the expression of Th2 cytokines viz. IL 4, IL10, TGF b and IL6.

REFERENCES

  1. Albers, R., Antoine, J.M., Bourdet- Sicard, R, Calder, P.C., Gleeson, M., Lesourd, B., Samarti N.S., Sanderson, I.R., Van L.J., Dias F. W.V and Watzl. B., (2005) Markers to measure immunomodulatijon in himan nutrition intervention studies. Br J Nutr 94:452-481.
  2. Brisbin,J.T., Zhou, H., Gong, J., Sabour, P., Akbari, M.R., Haghighi, H.R., Yu, H., Clarke, A., Sarson, A.J. and Sharif, S. (2007).Gene expression profiling of chicken lymphoid cells after treatment with Lactobacillus acidophilus cellular components. Developmental and Comparative Immunology 32: 563-574.
  3. Brisbin, J.T., Gong, J., Parvizi, P. and Sharif, S. (2010). Effects of Lactobacilli on Cytokine Expression by Chicken Spleen and Cecal Tonsil Cells. Clinical and vaccine immunology. 17(9):1337–1343.
  4. Castellazzi, A.M., Valsecchi, C., Montagna, L., Malfa, P., Ciprandi, G., Avanzini, M.A. andMarseglia, G.L. (2007). In vitro activation of mononuclear cells by two probiotics: Lactobacillus paracasei I 1688, Lactobacillus salivarius I 1794, and their mixture (PSMIX).Immunological Investigations. 36(4): 413-21.
  5. Dong, H., Rowland, I. and Yaqoob, P. (2011). Comparatives effects of six probiotic strains on immune function in vitro. British Journal of Nutrition, 108: 459-470.
  6. Donkor, O.N., Ravikumar,M., Proudfoot,O., Day,S.L., Apostolopoulos,V., Paukovics,G., Vasiljevic, T., Nutt, S.L. and Gill, H. (2012). Cytokine profile and induction of T helper type 17 and regulatory T cells by human peripheral mononuclear cells after microbial exposure.Clinical and Experimental Immunology, 167: 282–295.
  7. Drago L, Nicola L, Iemoli E, Banfi G and Vecci E D. (2010). Strain-dependent release of cytokines modulated by Lactobacillus salivarius human isolates in an in vitro model. BMC Res Notes 3, 44–48.
  8. Fuller, R. (1989).A Review: Probiotics in man and animals. Journal of Applied Bacteriology 66: 365-378.
  9. Hassan pyar.andPeh, K.K. (2014).Characterization and identification of lactobacillus acidophilus using biolog rapid identification system.International Journal of Pharmacy and Pharmaceutical Sciences, 6(1), 189-193.
  10. Hanan, T., Abbas, H., Anu, S., Juha, A. and Per, E.J. Saris. (2007). Identification of the Most Abundant Lactobacillus Species in the Crop of 1- and 5-Week-Old Broiler Chickens. Applied and Environmental. Microbiology. 73(24): 7867–7873
  11. Hanamanta, N., and Narayana, S. M. (2010). Immune responses in broiler chickens supplemented with prebiotic, probiotic, their combination and g-probiotic spl. Indian J. Anim. Res., 44 (2) : 150 – 152
  12. Hong, Y.H., Lillehoj, H.S., Lee, S.H., Dalloul, R.A. and Lillehoj, E.P. (2006).Analysis of chicken cytokine and chemokine gene expression following Eimeriaacervulina and Eimeriatenella infections. Veterinary Immunology and Immunopathology 114: 209-223.
  13. Izquierdo, M.E., Ennahar, S. and Sanz, Y. (2007). Differential immunomodulatory properties of Bifidobacteriumlogum strains: relevance to probiotic selection and clinical applications. Clinical and Experimental Immunology 150(3): 531–538.
  14. Lee., Na-Kyoung., Cheol-Won Yun., SeungWook Kim., Hyo-Ihl Chang., Chang-Won Kang.and Hyun-Dong Paik. (2008).Screening of Lactobacilli Derived from Chicken Feces and Partial Characterization of Lactobacillus acidophilus A12 as Animal Probiotics. Journal of Microbioogy and Biotechnology. 18: 338–342
  15. Ling, W.H., Saxelin, M., Hanninen, O. and Salminen, S. (1994).Enzyme Profile of Lactobacillus Strain GG by a Rapid API ZYM System: A Comparison of Intestinal Bacterial Strains. Microbial Ecology in Health and Diseases 7: 99-104.
  16. López P, Gueimonde M, Margolles A, Suárez A. (2010). Distinct Bifidobacterium strains drive different immune responses in vitro. Int J Food Microbiol 138:157–165.
  17. Medina, M., Izquierdo, E., Ennahar, S. and Sanz, Y. (2007) .Differential immunomodulatory properties of Bifidobacteriumlongum strains: relevance to probiotic selection and clinical applications. Clinical and Experimental Immunology 150: 531–538.
  18. Nang, N.T., Lee, J.S., Song, B.M., Kang, Y.M., Kim, H.S. and Seo, S.H. (2011). Induction of inflammatory cytokines and toll – like receptors in chicken infected with avian H9N2 influenza virus. Veterinary Research 42: 64.
  19. Nazila ,A., Enayatollah, K., Helen, P., Nassimeh, S. and Gholamreza, S. J.(2016). Selection and characterization of potential probiotic Lactobacilli spp isolated from chicken feces may be used as a potent antibacterial agent. Asian J. Dairy & Food Res, 35(1): 50-57
  20. Ram, N., Singh, D.P., Shailesh, B.S., and Paswan, V.K. (2012). Feed supplements vis-à-vis safe food through animal husbandry: a review. Agri. Reviews, 33 (4) : 315-321.
  21. Tamilvanan, T., Thiagarajan, M., Ramesh V., and Sivakumar, T.(2001). Performance of broiler chicken under floor system of management fed with different processed feed and probiotics. Indian J. Anim. Res., 35 (2): 88 - 91
  22. Tinrat, S., Saraya, S. and Chomnawang, M.T. (2011). Isolation and characterization of Lactobacillus salivarius MTC 1026 as a potential probiotic. Journal of.Generaland.Applied.Microbiology. 57: 365-378.
  23. Walter, J., Tannock, G.W., Tilsala-Timisjarvi,A., Rodtong,S., Loach,D.M., Munro,K. and Alatossava,T. (2000). Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species-specific PCR primers. Appl. Environ. Microbiol.66: 297–303 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 5 (may 2019) : 655-660,   Doi: 10.18805/ijar.B-3547

    In vitro selection of potential immunomodulatory multispecies lactic acid bacteria in broilers

    K
    K. Divya Manjari
    G
    G. Divya
    T
    T.S. Saravanan
    M
    M. Parthiban
    1Department of Animal Biotechnology, Madras Veterinary College, Faculty of Basic Sciences, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 007, Tamil Nadu, India.
    Cite article:- Manjari Divya K., Divya G., Saravanan T.S., Parthiban M. (2018). In vitro selection of potential immunomodulatory multispecies lactic acid bacteria in broilers. Indian Journal of Animal Research. 53(5): 655-660. doi: 10.18805/ijar.B-3547.

    ABSTRACT

    Modulation of host immunity is one of the proposed benefits of the consumption of probiotics.  In this study a total of 26 lactic acid bacteria belonging to eight different species were isolated from chicken gut. These twenty six probiotic organisms were characterized by Gram’s staining, biochemical methods using 21 sugars, pH tolerance and bile salt tolerance test.  These probiotic organisms were identified using PCR amplification of 16S-23S spacer region and sequencing analysis. Based on the sequencing data, 6 different species were found to exist namely Enterococcus faecalis; Enterococcus faecium, Enterococcus hirae, Weissella cibaria, Lactobacillus salivarius, Pediococcus acidilactici. To select the immunomodulatory probiotic bacteria, the scoring index was established based on different chicken cytokine expression and B and T cell receptor expression studies. According to scoring index, Pediococcus acidilactici, Lactobacillus salivarius and Weissellacibaria showed highest levels of fold increase in the expression of Th1 cytokines viz. IL2, IL 12p40, IL1b and TNF a. Enterococcus faecalis and Enterococcus faecium showed highest levels of fold increase in the expression of Th2 cytokines viz. IL 4, IL10, TGF b and IL6.

    REFERENCES

    1. Albers, R., Antoine, J.M., Bourdet- Sicard, R, Calder, P.C., Gleeson, M., Lesourd, B., Samarti N.S., Sanderson, I.R., Van L.J., Dias F. W.V and Watzl. B., (2005) Markers to measure immunomodulatijon in himan nutrition intervention studies. Br J Nutr 94:452-481.
    2. Brisbin,J.T., Zhou, H., Gong, J., Sabour, P., Akbari, M.R., Haghighi, H.R., Yu, H., Clarke, A., Sarson, A.J. and Sharif, S. (2007).Gene expression profiling of chicken lymphoid cells after treatment with Lactobacillus acidophilus cellular components. Developmental and Comparative Immunology 32: 563-574.
    3. Brisbin, J.T., Gong, J., Parvizi, P. and Sharif, S. (2010). Effects of Lactobacilli on Cytokine Expression by Chicken Spleen and Cecal Tonsil Cells. Clinical and vaccine immunology. 17(9):1337–1343.
    4. Castellazzi, A.M., Valsecchi, C., Montagna, L., Malfa, P., Ciprandi, G., Avanzini, M.A. andMarseglia, G.L. (2007). In vitro activation of mononuclear cells by two probiotics: Lactobacillus paracasei I 1688, Lactobacillus salivarius I 1794, and their mixture (PSMIX).Immunological Investigations. 36(4): 413-21.
    5. Dong, H., Rowland, I. and Yaqoob, P. (2011). Comparatives effects of six probiotic strains on immune function in vitro. British Journal of Nutrition, 108: 459-470.
    6. Donkor, O.N., Ravikumar,M., Proudfoot,O., Day,S.L., Apostolopoulos,V., Paukovics,G., Vasiljevic, T., Nutt, S.L. and Gill, H. (2012). Cytokine profile and induction of T helper type 17 and regulatory T cells by human peripheral mononuclear cells after microbial exposure.Clinical and Experimental Immunology, 167: 282–295.
    7. Drago L, Nicola L, Iemoli E, Banfi G and Vecci E D. (2010). Strain-dependent release of cytokines modulated by Lactobacillus salivarius human isolates in an in vitro model. BMC Res Notes 3, 44–48.
    8. Fuller, R. (1989).A Review: Probiotics in man and animals. Journal of Applied Bacteriology 66: 365-378.
    9. Hassan pyar.andPeh, K.K. (2014).Characterization and identification of lactobacillus acidophilus using biolog rapid identification system.International Journal of Pharmacy and Pharmaceutical Sciences, 6(1), 189-193.
    10. Hanan, T., Abbas, H., Anu, S., Juha, A. and Per, E.J. Saris. (2007). Identification of the Most Abundant Lactobacillus Species in the Crop of 1- and 5-Week-Old Broiler Chickens. Applied and Environmental. Microbiology. 73(24): 7867–7873
    11. Hanamanta, N., and Narayana, S. M. (2010). Immune responses in broiler chickens supplemented with prebiotic, probiotic, their combination and g-probiotic spl. Indian J. Anim. Res., 44 (2) : 150 – 152
    12. Hong, Y.H., Lillehoj, H.S., Lee, S.H., Dalloul, R.A. and Lillehoj, E.P. (2006).Analysis of chicken cytokine and chemokine gene expression following Eimeriaacervulina and Eimeriatenella infections. Veterinary Immunology and Immunopathology 114: 209-223.
    13. Izquierdo, M.E., Ennahar, S. and Sanz, Y. (2007). Differential immunomodulatory properties of Bifidobacteriumlogum strains: relevance to probiotic selection and clinical applications. Clinical and Experimental Immunology 150(3): 531–538.
    14. Lee., Na-Kyoung., Cheol-Won Yun., SeungWook Kim., Hyo-Ihl Chang., Chang-Won Kang.and Hyun-Dong Paik. (2008).Screening of Lactobacilli Derived from Chicken Feces and Partial Characterization of Lactobacillus acidophilus A12 as Animal Probiotics. Journal of Microbioogy and Biotechnology. 18: 338–342
    15. Ling, W.H., Saxelin, M., Hanninen, O. and Salminen, S. (1994).Enzyme Profile of Lactobacillus Strain GG by a Rapid API ZYM System: A Comparison of Intestinal Bacterial Strains. Microbial Ecology in Health and Diseases 7: 99-104.
    16. López P, Gueimonde M, Margolles A, Suárez A. (2010). Distinct Bifidobacterium strains drive different immune responses in vitro. Int J Food Microbiol 138:157–165.
    17. Medina, M., Izquierdo, E., Ennahar, S. and Sanz, Y. (2007) .Differential immunomodulatory properties of Bifidobacteriumlongum strains: relevance to probiotic selection and clinical applications. Clinical and Experimental Immunology 150: 531–538.
    18. Nang, N.T., Lee, J.S., Song, B.M., Kang, Y.M., Kim, H.S. and Seo, S.H. (2011). Induction of inflammatory cytokines and toll – like receptors in chicken infected with avian H9N2 influenza virus. Veterinary Research 42: 64.
    19. Nazila ,A., Enayatollah, K., Helen, P., Nassimeh, S. and Gholamreza, S. J.(2016). Selection and characterization of potential probiotic Lactobacilli spp isolated from chicken feces may be used as a potent antibacterial agent. Asian J. Dairy & Food Res, 35(1): 50-57
    20. Ram, N., Singh, D.P., Shailesh, B.S., and Paswan, V.K. (2012). Feed supplements vis-à-vis safe food through animal husbandry: a review. Agri. Reviews, 33 (4) : 315-321.
    21. Tamilvanan, T., Thiagarajan, M., Ramesh V., and Sivakumar, T.(2001). Performance of broiler chicken under floor system of management fed with different processed feed and probiotics. Indian J. Anim. Res., 35 (2): 88 - 91
    22. Tinrat, S., Saraya, S. and Chomnawang, M.T. (2011). Isolation and characterization of Lactobacillus salivarius MTC 1026 as a potential probiotic. Journal of.Generaland.Applied.Microbiology. 57: 365-378.
    23. Walter, J., Tannock, G.W., Tilsala-Timisjarvi,A., Rodtong,S., Loach,D.M., Munro,K. and Alatossava,T. (2000). Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species-specific PCR primers. Appl. Environ. Microbiol.66: 297–303 
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