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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 52 issue 12 (december 2018) : 1739-1744,   Doi: 10.18805/ijar.B-3429

Isolation and screening of Lactobacillus species from dogs for probiotic action
 

N
N. Mohammad Sharif
B
B. Sreedevi
R
R.K. Chaitanya
D
D. Sreenivasulu
1Department of Veterinary Microbiology, College of Veterinary Science, Sri Venkateswara Veterinary University, Tirupati-517 502, Andhra Pradesh, India.
Cite article:- Sharif Mohammad N., Sreedevi B., Chaitanya R.K., Sreenivasulu D. (2017). Isolation and screening of Lactobacillus species from dogs for probiotic action. Indian Journal of Animal Research. 52(12): 1739-1744. doi: 10.18805/ijar.B-3429.

ABSTRACT

The present study aimed at the detection of suitable Lactobacillus species from dogs for usage as probiotic. A total of 67 rectal swabs from healthy pups were analyzed and 49 (73.1%) Lactobacillus isolates were identified based on morphological, biochemical characteristics and confirmed by genus specific PCR targeting 16S rRNA gene. A total of 20 isolates that showed strong aggregation, high cell surface hydrophobicity, acid and bile tolerance were screened for in-vitro antibacterial activity by agar well diffusion assay, where prominent inhibitory zones were observed against majority of the test pathogens. Reduction in antibacterial activity was noticed after neutralization, proteinase K and heat treatment of supernatants. Partial 16S rRNA nucleotide sequence analysis of seven Lactobacillus isolates that showed prominent in-vitro antibacterial activity revealed maximum sequence homology with Lactobacillus fermentum (for six isolates) and Lactobacillus agilis (for one isolate). The in-vitro antibacterial activity results highlighted the need for in-vivo studies in India to establish probiotic potential of canine faecal Lactobacillus species in the near future.

REFERENCES

  1. Baillon, M. L. A., Marshall-Jones, Z. V. and Butterwick, R. F. (2004). Effects of probiotic Lactobacillus acidophilus strain DSM13241 in healthy adult dogs. Am. J. Vet. Res. 65(3):338-343.
  2. Beasley, S. S., Manninen, T. J. K. and Saris, P. E. J. (2006). Lactic acid bacteria isolated from canine faeces. J. Appl. Microbiol. 101(1):131-    138.
  3. Benno, Y., Nakao, H., Uchida, K. and Mitsuoka, T. (1992). Impact of the advances in age on the gastrointestinal microflora of beagle dogs. J. Vet. Med. Sci. 54(4):703-706.
  4. Bilkova, A., Sepova, H. K., Bukovsky, M. and Bezakova, L. (2011). Antibacterial potential of lactobacilli isolated from a lamb. Vet. Med. 56(56): 319-324.
  5. Ehrmann, M.A., Kurzak, P., Bauer, J. and Vogel, R.F. (2002). Characterization of lactobacilli towards their use as probiotic adjuncts in poultry. J. Appl. Microbiol. 92(5): 966-975.
  6. Garcia-Mazcorro, J. F., Lanerie, D. J., Dowd, S. E., Paddock, C. G., Grutzner, N., et al.,(2011). Effect of a multi-species synbiotic formulation on fecal bacterial microbiota of healthy cats and dogs as evaluated by pyrosequencing. FEMS Microbiol. Ecol. 78(3):542-554.
  7. Greetham, H. L., Giffard, C., Hutson, R. A., Collins, M. D. and Gibson, G. R. (2002). Bacteriology of the Labrador dog gut: a cultural and genotypic approach. J. Appl. Microbiol. 93(4):640-646.
  8. Khalil, R., El-Halafawy, K., Mahrous, H., Kamaly, K., Frank, J. and El Soda, M. (2007). Evaluation of the probiotic potential of lactic acid bacteria isolated from faeces of breast-fed infants in Egypt. Afr. J. Biotechnol. 6(7): 939-949.
  9. Kumar, S., Pattanaik, A.K., Sharma, S. and Jadhav, S.E. (2016). Species-specific probiotic Lactobacillus johnsonii CPN23 supplementation modulates blood biochemical profile and erythrocytic antioxidant indices in Labrador dogs. Indian J. Anim. Sci. 86(8).
  10. Kumar, S., Pattanaik, A.K., Sharma, S., Jadhav, S.E., Dutta, N. and Kumar, A. (2017). Probiotic Potential of a Lactobacillus Bacterium of Canine Faecal-Origin and Its Impact on Select Gut Health Indices and Immune Response of Dogs. Probiotics Antimicrob. Proteins, 1-16.
  11. Martin, R., Olivares, M., Perez, M., Xaus, J., Torre, C., Fernandez, L. and Rodriguez, J. M. (2010). Identification and evaluation of the probiotic potential of lactobacilli isolated from canine milk. Vet. J. 185(2): 193-198.
  12. McCoy, S. and Gilliland, S. E. (2007). Isolation and characterization of Lactobacillus species having potential for use as probiotic cultures for dogs. J. Food. Sci., 72(3):M94-M97.
  13. Nair, P. S. (2000). Studies on lactic acid bacteria from tropical fish and shell fish. Ph.D. Thesis submitted to Central institute of fisheries technology, Cochin University of Science and Technology, Cochin, India - 682022.
  14. Oyetayo, V. O. and Oyetayo, F. L. (2005). Potential of probiotics as biotherapeutic agents targeting the innate immune system. Afr. J. Biotechnol.4 (2):123-127.
  15. Perelmuter, K., Fraga, M. and Zunino, P. (2008). In vitro activity of potential probiotic Lactobacillus murinus isolated from the dog. J. Appl. Microbiol. 104(6):1718-1725.
  16. Sanders, M. E. (2008). Probiotics: definition, sources, selection, and uses. Clin. Infect. Dis. 46: S58-S61.
  17. Silva, B. C., Jung, L. R. C., Sandes, S. H. C., Alvim, L. B., Bomfim, M. R. Q et al., (2013). In vitro assessment of functional properties of lactic acid bacteria isolated from faecal microbiota of healthy dogs for potential use as probiotics. Benef. Microbes. 4(3):267-275.
  18. Sneath, P. H. A. and Holt, J. G. (2001). Bergey’s Manual of Systematic Bacteriology, 2nd edition. VOl 1, A Waverly Company, Williams & Wilkins, Springer-Verlag, NY, USA.
  19. Strompfova, V. and Laukova, A. (2014). Isolation and characterization of faecal bifidobacteria and lactobacilli isolated from dogs and primates. Anaerobe, 29:108-112.
  20. Taheri, H.R., Moravej, H., Tabandeh, F., Zaghari, M. and Shivazad, M. (2009). Screening of lactic acid bacteria toward their selection as a source of chicken probiotic. Poult. Sci. 88(8): 1586-1593.
  21. Tang, Y., Manninen, T. J., and Saris, P. E. (2012). Dominance of Lactobacillus acidophilus in the facultative jejunal Lactobacillus microbiota of fistulated beagles. Appl. Environ. Microbiol. 78(19):7156-7159.
  22. Vineetha, P.G., Tomar, S., Saxena, V.K., Sandeep, S., Adil, K. and Mukesh, K. (2016). Screening of Lactobacillus isolates from gastrointestinal tract of guinea fowl for probiotic qualities using in-vitro tests to select species-specific probiotic candidates. Br. Poult. Sci. 57(4): 474-482.
  23. Wani, S.A., Samanta, I., Munshi, Z.H., Bhat, M.A. and Nishikawa, Y. (2006). Shiga toxin producing Escherichia coli and enteropathogenic Escherichia coli in healthy goats in India: occurrence and virulence properties. J. Appl. Microbiol.100: 108-113. 
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Indian Journal of Animal Research
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    Research Article

    volume 52 issue 12 (december 2018) : 1739-1744,   Doi: 10.18805/ijar.B-3429

    Isolation and screening of Lactobacillus species from dogs for probiotic action
     

    N
    N. Mohammad Sharif
    B
    B. Sreedevi
    R
    R.K. Chaitanya
    D
    D. Sreenivasulu
    1Department of Veterinary Microbiology, College of Veterinary Science, Sri Venkateswara Veterinary University, Tirupati-517 502, Andhra Pradesh, India.
    Cite article:- Sharif Mohammad N., Sreedevi B., Chaitanya R.K., Sreenivasulu D. (2017). Isolation and screening of Lactobacillus species from dogs for probiotic action. Indian Journal of Animal Research. 52(12): 1739-1744. doi: 10.18805/ijar.B-3429.

    ABSTRACT

    The present study aimed at the detection of suitable Lactobacillus species from dogs for usage as probiotic. A total of 67 rectal swabs from healthy pups were analyzed and 49 (73.1%) Lactobacillus isolates were identified based on morphological, biochemical characteristics and confirmed by genus specific PCR targeting 16S rRNA gene. A total of 20 isolates that showed strong aggregation, high cell surface hydrophobicity, acid and bile tolerance were screened for in-vitro antibacterial activity by agar well diffusion assay, where prominent inhibitory zones were observed against majority of the test pathogens. Reduction in antibacterial activity was noticed after neutralization, proteinase K and heat treatment of supernatants. Partial 16S rRNA nucleotide sequence analysis of seven Lactobacillus isolates that showed prominent in-vitro antibacterial activity revealed maximum sequence homology with Lactobacillus fermentum (for six isolates) and Lactobacillus agilis (for one isolate). The in-vitro antibacterial activity results highlighted the need for in-vivo studies in India to establish probiotic potential of canine faecal Lactobacillus species in the near future.

    REFERENCES

    1. Baillon, M. L. A., Marshall-Jones, Z. V. and Butterwick, R. F. (2004). Effects of probiotic Lactobacillus acidophilus strain DSM13241 in healthy adult dogs. Am. J. Vet. Res. 65(3):338-343.
    2. Beasley, S. S., Manninen, T. J. K. and Saris, P. E. J. (2006). Lactic acid bacteria isolated from canine faeces. J. Appl. Microbiol. 101(1):131-    138.
    3. Benno, Y., Nakao, H., Uchida, K. and Mitsuoka, T. (1992). Impact of the advances in age on the gastrointestinal microflora of beagle dogs. J. Vet. Med. Sci. 54(4):703-706.
    4. Bilkova, A., Sepova, H. K., Bukovsky, M. and Bezakova, L. (2011). Antibacterial potential of lactobacilli isolated from a lamb. Vet. Med. 56(56): 319-324.
    5. Ehrmann, M.A., Kurzak, P., Bauer, J. and Vogel, R.F. (2002). Characterization of lactobacilli towards their use as probiotic adjuncts in poultry. J. Appl. Microbiol. 92(5): 966-975.
    6. Garcia-Mazcorro, J. F., Lanerie, D. J., Dowd, S. E., Paddock, C. G., Grutzner, N., et al.,(2011). Effect of a multi-species synbiotic formulation on fecal bacterial microbiota of healthy cats and dogs as evaluated by pyrosequencing. FEMS Microbiol. Ecol. 78(3):542-554.
    7. Greetham, H. L., Giffard, C., Hutson, R. A., Collins, M. D. and Gibson, G. R. (2002). Bacteriology of the Labrador dog gut: a cultural and genotypic approach. J. Appl. Microbiol. 93(4):640-646.
    8. Khalil, R., El-Halafawy, K., Mahrous, H., Kamaly, K., Frank, J. and El Soda, M. (2007). Evaluation of the probiotic potential of lactic acid bacteria isolated from faeces of breast-fed infants in Egypt. Afr. J. Biotechnol. 6(7): 939-949.
    9. Kumar, S., Pattanaik, A.K., Sharma, S. and Jadhav, S.E. (2016). Species-specific probiotic Lactobacillus johnsonii CPN23 supplementation modulates blood biochemical profile and erythrocytic antioxidant indices in Labrador dogs. Indian J. Anim. Sci. 86(8).
    10. Kumar, S., Pattanaik, A.K., Sharma, S., Jadhav, S.E., Dutta, N. and Kumar, A. (2017). Probiotic Potential of a Lactobacillus Bacterium of Canine Faecal-Origin and Its Impact on Select Gut Health Indices and Immune Response of Dogs. Probiotics Antimicrob. Proteins, 1-16.
    11. Martin, R., Olivares, M., Perez, M., Xaus, J., Torre, C., Fernandez, L. and Rodriguez, J. M. (2010). Identification and evaluation of the probiotic potential of lactobacilli isolated from canine milk. Vet. J. 185(2): 193-198.
    12. McCoy, S. and Gilliland, S. E. (2007). Isolation and characterization of Lactobacillus species having potential for use as probiotic cultures for dogs. J. Food. Sci., 72(3):M94-M97.
    13. Nair, P. S. (2000). Studies on lactic acid bacteria from tropical fish and shell fish. Ph.D. Thesis submitted to Central institute of fisheries technology, Cochin University of Science and Technology, Cochin, India - 682022.
    14. Oyetayo, V. O. and Oyetayo, F. L. (2005). Potential of probiotics as biotherapeutic agents targeting the innate immune system. Afr. J. Biotechnol.4 (2):123-127.
    15. Perelmuter, K., Fraga, M. and Zunino, P. (2008). In vitro activity of potential probiotic Lactobacillus murinus isolated from the dog. J. Appl. Microbiol. 104(6):1718-1725.
    16. Sanders, M. E. (2008). Probiotics: definition, sources, selection, and uses. Clin. Infect. Dis. 46: S58-S61.
    17. Silva, B. C., Jung, L. R. C., Sandes, S. H. C., Alvim, L. B., Bomfim, M. R. Q et al., (2013). In vitro assessment of functional properties of lactic acid bacteria isolated from faecal microbiota of healthy dogs for potential use as probiotics. Benef. Microbes. 4(3):267-275.
    18. Sneath, P. H. A. and Holt, J. G. (2001). Bergey’s Manual of Systematic Bacteriology, 2nd edition. VOl 1, A Waverly Company, Williams & Wilkins, Springer-Verlag, NY, USA.
    19. Strompfova, V. and Laukova, A. (2014). Isolation and characterization of faecal bifidobacteria and lactobacilli isolated from dogs and primates. Anaerobe, 29:108-112.
    20. Taheri, H.R., Moravej, H., Tabandeh, F., Zaghari, M. and Shivazad, M. (2009). Screening of lactic acid bacteria toward their selection as a source of chicken probiotic. Poult. Sci. 88(8): 1586-1593.
    21. Tang, Y., Manninen, T. J., and Saris, P. E. (2012). Dominance of Lactobacillus acidophilus in the facultative jejunal Lactobacillus microbiota of fistulated beagles. Appl. Environ. Microbiol. 78(19):7156-7159.
    22. Vineetha, P.G., Tomar, S., Saxena, V.K., Sandeep, S., Adil, K. and Mukesh, K. (2016). Screening of Lactobacillus isolates from gastrointestinal tract of guinea fowl for probiotic qualities using in-vitro tests to select species-specific probiotic candidates. Br. Poult. Sci. 57(4): 474-482.
    23. Wani, S.A., Samanta, I., Munshi, Z.H., Bhat, M.A. and Nishikawa, Y. (2006). Shiga toxin producing Escherichia coli and enteropathogenic Escherichia coli in healthy goats in India: occurrence and virulence properties. J. Appl. Microbiol.100: 108-113. 
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