Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Isolation, In vitro Antibacterial Activity, Bacterial Sensitivity and Plasmid Profile of Lactobacilli

  • Lonkar, P. (Department of Microbiology, Nagpur Veterinary College) ;
  • Harne, S.D. (Department of Microbiology, Nagpur Veterinary College) ;
  • Kalorey, D.R. (Department of Microbiology, Nagpur Veterinary College) ;
  • Kurkure, N.V. (Department of Pathology, Nagpur Veterinary College)
  • Received : 2004.11.25
  • Accepted : 2005.04.07
  • Published : 2005.09.01
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Abstract

The present research work was conducted to evaluate the beneficial effects as well as the safety aspects of lactobacilli as probiotic. Lactobacilli were isolated from poultry faecal samples, feed samples and from some known preparations procured from poultry feed manufacturers. L. acidophilus and L. sporogenes were tested for the antibacterial activity against four poultry pathogens viz. Escherichia coli, Salmonella spp., Proteus spp. and Pseudomonas aeruginosa. Cell free supernatant (CFS) of L. acidophilus exhibited significantly higher antibacterial activity against Salmonella spp. at original pH (4.50${\pm}$0.02). At the adjusted pH (6.50${\pm}$0.02) significantly higher antibacterial activity was recorded against indicator organism except for P. aeruginosa. Likewise, L. sporogenes exhibited similar antibacterial activity at original as well as adjusted pH except for E. coli. Antibacterial activity against E. coli was significantly higher at adjusted pH than at original pH of CFS. The competitive exclusion of E. coli by lactobacilli over the intestinal epithelial cells (IEC) was checked. L. acidophilus strain I, which was of poultry origin, exhibited maximum attachment over IEC as compared to other three strains of non-poultry origin viz. L. acidophilus strain II, L. sporogenes strain I and II. Overall, L. acidophilus exhibited higher competitive exclusion as compared to L. sporogenes. All the lactobacilli of poultry origin were most sensitive to penicillin G, amoxycillin, ampicillin and chloramphenicol, least sensitive to sulphamethizole, ciprofloxacin, neomycin, norfloxacin and pefloxacin and resistant to metronidazole and nalidixic acid. The isolates from probiotic preparations were most sensitive to ampicillin, amoxycillin and tetracycline, least sensitive to sulphamethizole, norfloxacin, neomycin and ceftriazone and resistant to nalidixic acid and metronidazole. Eight of the multiple drug resistant lactobacilli isolates were studied for the presence of plasmids. Plasmids could be extracted from six isolates of lactobacilli. These plasmids could be responsible for bacteriocin production or for antibiotic resistance of the strains. The lactobacilli need further studies regarding their safety for use in the probiotic preparations.

Keywords

References

  1. Ahn, Y. T., K. L. Lim, J. C. Ryn, D. K. Kang, J. S. Ham, Y. H. Jang and H. U. Kim. 2002. Characterization of Lactobacillus acidophilus isolated from piglets and chicken. Asian-Aust. J. Anim. Sci. 15:1790-1797.
  2. Bae, H. C., M. S. Nam and J. Y. Lee. 2002. Probiotic characterization of acid and bile tolerant Lactobacillus salivarius subsp. salivarius from Korean faeces. Asian-Aust. J. Anim. Sci. 15:1798-1807.
  3. Bae, J. S., J. R. Byun and Y. H. Yoon. 2003. In vivo antagonistic effect of Lactobascillus helveticus CU 631 against Salmonella enteritidis KU 101 infection. Asian-Aust. J. Anim. Sci. 16:430-434.
  4. Bauer, A. W., W. M. M. Kirby, J. C. Shernis and M. Turek. 1966. Antibiotic susceptibility testing by a standardized single disc method. Am. J. Clin. Pathol. 45:493-496.
  5. Byun, J. R., Y. J. Baik and Y. H. Yoon. 2004. Effects of feeding Lactobacillus spp. on the level of cell glutathione sulphydryl and immunoglobulins M in ICR mice. Asian-Aust. J. Anim. Sci. 17:415-419.
  6. Charteris, W. P., P. Kelly, Lorenzo Morelli and K. J. Collins. 1998. Antibiotic susceptibility of potentially probiotic Lactobacillus species. J. Food Protect. 61:1636-1643.
  7. Clewell, D. B., Y. Yagi, G. M. Dunny and S. K. Schultz. 1974. Characterization of three deoxyribonucleic acid molecules in a strain of Streptococcus faecalis. Identification of a plasmid determining erythromycin resistance. J. Bacteriol. 177:283-289.
  8. Cocconcelli, P. S., L. Morelli and M. Vescovo. 1985. Conjugal transfer of antibiotic resistances from Lactobacillus to Streptococcus lactis. Microbiologie-Ailments-Nutrition 3:163-165.
  9. Ehrmann, M. A., P. Kurzak, J. Bauer and R. F. Vogel. 2002. Characterization of lactobacilli towards their use as probiotic adjuncts in poultry. J. Appl. Microbiol. 92:966-975.
  10. Frere, J. 1994. Simple method for extracting plasmid DNA from lactic acid bacteria. Lett. Appl. Microbiol. 18:227-229.
  11. Fuller, R. 1978. Epithelial attachment and other factors controlling the colonization of the intestine of the gnotobiotic chicken by lactobacilli. J. Appl. Bacteriol. 45:389-395.
  12. Gao, W. and Q. X. Meng. 2004. Lectin activity and chemical characteristic of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. from gastrointestinal mucosa of growing pigs. Asian-Aust. J. Anim. Sci. 17: 863-868.
  13. Gibson, E.M., N.H. Chace, S.B. London and J. London. 1979. Transfer of plasmid mediated antibiotic resistance from streptococci to lactobacilli. J. Bacteriol. 137:614-619.
  14. Gilliland, S. E., M. L. Speck and C. G. Morgan. 1975. Detection of Lactobacillus acidophilus in faeces of humans, pigs and chickens. Appl. Microbiol. 30:541-545.
  15. Hakkinen, M. and C. Schneitz. 1999. Efficacy of a commercial competitive exclusion product against Campylobacter jejuni. Br. Poult. Sci. 40:619-621.
  16. Jeffrey, J. S. 1999. Use of competitive exclusion products for poultry. Poultry Fact Sheet No. 30. Co-operative extension, University of California.
  17. Jin, L. Z., Y. W. Ho, N. Abdullah and S. Jalaluddin. 1996a. Influence of dried Bacillus subtilis and lactobacilli cultures on intestinal microflora and performance in broilers. Asian-Aust. J. Anim. Sci. 9:397-404.
  18. Jin, L. Z., Y. W. Ho, M. A. Ali, N. Abbullah and S. Galalubin. 1996b. Effect of adherent Lactobacillus spp. on in vitro adherence of salmonellae to the intestinal epithelial cells of chicken. J. Appl. Bacteriol. 81:201-206.
  19. Kandler, O. and N. Weiss. 1986. Genus Lactobacillus. In: Bergey’s Manual of Systematic Bacteriology, Vol. 2 (Ed. P. H. A. Sneath, N. S. Mair, M. E. Sharpe and J. G. Holt). Williams and Wilkins, Baltimore. pp. 1209-1234.
  20. Katla, A. K., H. Kruse, G. Johnsen and H. Herikstad. 2001. Antimicrobial susceptibility of starter culture bacteria used in Norwegian dairy products. Inter. J. Food Microbiol. 67:147-152.
  21. Klein, G., C. Hallmann, I. A. Casas, J. Abad, J. Louwers and G. Reuter. 2000. Exclusion of Van A, Van B and Van C type glycopeptide resistance in strains of Lactobacillus reuteri and L. rhamnosus used as probiotics by polymerase chain reaction and hybridization methods. J. Appl. Microbiol. 89:815-821.
  22. Lebenthal, E. and Y. Lebenthal. 2002. Probiotics-An important therapeutic concept awaiting validation. Indian Med. Assoc. J. 4:374-375.
  23. Luchansky, J. B., E. G. Kleeman, R. R. Raya and T. R. Klaenhammer. 1989. Genetic transfer systems for delivery of plasmid deoxyribonucleic acid to Lactobacillus acidophilus. ADH:conjugation, electoproporation and transduction. J. Dairy Sci. 72:1408-17.
  24. Mayara-Makinen, A., M. Manninen and H. Gynenberg. 1983. The adherence of lactic acid bacteria to the epithelial cells of pigs and calves. J. Appl. Bacteriol. 55:241-245.
  25. Miyamoto, T., T. Horie, T. Fujiwara, T. Fukata, K. Sasai and E. Baba. 2000. Lactobacillus flora in the cloaca and vagina of hens and its inhibitory activity against Salmonella enteritidis in vitro. Poult. Sci. 79:7-11.
  26. Morelli, L., P. G. Sarra and V. Bottazzi. 1988. In vivo transfer of pAMB1, from Lactobacillus reuteri to Enterococcus faecalis. J. Appl. Bacteriol. 65:371-375.
  27. Muralidhara, K., G. G. Sheggeby, P. R. Elliker, D. C. England and W. E. Sandine. 1977. Effect of feeding lactobacilli on the coliform and Lactobacillus flora of intestinal tissues and faeces from pigs. J. Food Protect. 10:288-295.
  28. Prasad, R. V. and D. N. Gandhi. 1987. Factors affecting the production of antibacterial substances in Lactobacillus acidophilus strain R. Indian J. Dairy Sci. 40:1-3.
  29. Rolfe, R. D. 2000. The role of probiotic cultures in the control of gastrointestinal health. J. Nutr. 130:3965-4025.
  30. Romero, D. A. and L. L. McKay. 1985. Isolation and plasmid characterization of Lactobacillus species in the manufacture of fermented sausage. J. Food Prot. 48:1028-1035.
  31. Schneitz, C., L. Nuotio and K. Lounatma. 1993. Adhesion of Lactobacillus acidophilus to avian intestinal epithelial cells mediated by the crystalline bacterial cell surface layer (Slayer). J. Appl. Bacteriol. 74:290-294.
  32. Snedecor, G. W. and W. G. Cochran. 1967. Statistical Methods, 6th Edn., Oxford and IBH Publishing Co., New Delhi.
  33. Surono, I. S. 2003. In vitro probiotic properties of indigenous dadih lactic acid bacteria. Asian-Aust. J. Anim. Sci. 16:726-731.
  34. Tannock, G. W. 1987. Conjugal transfer of plasmid pAMB1 in Lactobacillus reuteri and between lactobacilli and Enterococcus faecalis. Appl. Environ. Microbiol. 53:2693-2695.
  35. Todorikil, K., T. Mukai, S. Sato and T. Toba. 2001. Inhibition of adhesion of food born pathogens to Caco-2 cells by Lactobacillus strains. J. Appl. Microbiol. 91:154-159.
  36. Vescovo, M., L. Morelli, V. Bottazzi and M. J. Gasson. 1983. Conjugal transfer of broad host range plasmid pAMB1, into enteric species of lactic acid bacteria. Appl. Environ. Microbiol. 46:753-755.
  37. Weinack, O. M., G. H. Snoyenbos, C. F. Smyser and A. S. Soerjadi. 1981. Competitive exclusion of intestinal colonization of E. coli in chicks. Avian Dis. 25:695-705.
  38. Yoon, Y. H. and B. R. Won. 2002. Antigonism against Helicobacter pylori and proteolysis of Lactobacillus helveticus CU 631 and Strain Identification. Asian-Aust. J. Anim. Sci. 15:1057-1065.

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