Screening and Identification of Lactic Acid Bacteria Producing Antimicrobial Compounds from PIG Gastrointestinal Tracts
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Abstract
A total of 87 lactic acid bacteria were isolated from 15 samples of pig gastrointestinal tract. Antimicrobial activities of all isolates grown under oxygen-restricted conditions to eliminate the effect of hydrogen peroxide were tested against 5 pathogenic bacterial strains using agar well diffusion technique. Seventeen isolates exhibited antimicrobial activity against at least one indicator strain tested. Among the 17 isolates, two strains of Lactobacillus amylovorus/ L. sobrius and one strain of Lactobacillus reuteri were found to produce antimicrobial compounds other than organic acids. The neutralized supernatants from two L. amylovorus/ L. sobrius strains inhibited the growth of Escherichia coli and Listeria monocytogenes, respectively. Therefore, these strains may have potential use as an alternative to antibiotics for pigs. The strains can also be used to produce antimicrobial compounds which can be a substitute for chemical preservatives in food industry. Diversity of lactic acid bacteria with antimicrobial activities was studied by identification to the species level of the 17 isolates. This was achieved by sequencing of approximately 500 bp of 16S rRNA gene which contained V1 and V2 variable regions and comparing the sequences with existing data in GenBank. Of the 17 isolates, 7 isolateswere identified as Lactobacillus reuteri, the other 10 isolates were identified as Lactobacillus amylovorus/L. sobrius (4 isolates), Lactobacillus mucosae (3 isolates), Lactobacillus acidophilus johnsonii/L. johnsonii (2 isolates) and Streptococcus alactolyticus (1 isolate).
KEYWORDS: antimicrobial compounds, lactic acid bacteria, pig gastrointestinal tract
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References
[2] Pascual, M., Hugas, M., Badiola, J.I., Monfort, J.M. and Garriga, M. 1999 Lactobacillus salivarius CTC2197 Prevents Salmonella enteritidis Colonization in Chickens, Applied and Environmental Microbiology, 65(11), 4981-4986.
[3] Klein, G., Pack, A., Bonaparte, C. and Reuter, G. 1998 Taxonomy and Physiology of Probiotic Lactic Acid Bacteria: A Review, International Journal of Food Microbiology,41(2), 103-125.
[4] Chukeatirote, A. 2003 Potential Use of Probiotics, Songklanakarin Journal Science and Technology, 25(2), 275- 282.
[5] Casey, P.G., Casey, G.D., Gardiner, G.E., Tangney, M., Stanton, C., Ross, R.P., Hill, C. and Fitzgerald, G.F. 2004 Isolation and Characterization of Anti-Salmonella Lactic Acid Bacteria from the Porcine Gastrointestinal Tract, Letters in Applied Microbiology, 39(5),431-438.
[6] Brashears, M.M., Jaroni, D. and Trimble, J. 2003 Isolation, Selection, and Characterization of Lactic Acid Bacteria for a Competitive Exclusion Product to Reduce Shedding of Escherichia coli O157:H7 in Cattle, Journal of Food Protection, 66(3), 355-363.
[7] Kim, P., Min, J., Chang, Y.-H., Kim, S., Kim, S.-J. and Park, Y.-H. 2007 Probiotic Properties of Lactobacillus and Bifidobacterium Strains Isolated from Porcine Gastrointestinal Tract, Applied Microbiology and Biotechnology, 74, 1103-1111.
[8] Chaveerach, P., Lipman, L.J.A. and van Knapen, F. 2004 Antagonistic Activities of Several Bacteria on in vitro Growth of 10 Strains of Campylobacter jejuni/coli, International Journal of Food Microbiology, 90, 43-50.
[9] Mukai, T., Asasaka, T., Sato, E., Mori, K., Matsumoto, M. and Ohori, H. 2002 Inhibition of Binding of Helicobacter pylori to the Glycolipid Receptors by Probiotic Lactobacillus reuteri, FEMS Immunology and Medical Microbiology, 3, 105-110.
[10] Randazzo, C.L., Restuccia, C., Romano A.D. and Caggia, C. 2004 Lactobacillus casei, Dominant Species in Naturally Fermented Sicilian Green Olives, International Journal of Food Microbiology, 90, 9-14.
[11] Botina, S.G., Tsygankov, Yu, D. and Sukhodolets, V.V. 2006 Identification of Industrial Strains of Lactic Acid Bacteria by Methods of Molecular Genetic Typing, Russian Journal of Genetics, 42(12), 1367-1379.
[12] Kullen, M.J., Sanozky-Dawes, R.B., Crowell, D.C. and Klaenhammer, T.R. 2000 Use of the DNA Sequence of Variable Regions of the 16S rRNA Gene for Rapid and Accurate Identification of Bacteria in the Lactobacillus acidophilus Complex, Journal of Applied Microbiology, 89, 511-516.
[13] Tarnberg, M., Jakobsson, T., Jonasson, J. and Forsum, U. 2002 Identification of Randomly Selected Colonies of Lactobacilli from Normal Vaginal Fluid by Pyrosequencing of the 16S rDNA Variable V1 and V3 Regions, APMIS, 110, 802-810.
[14] Schillinger, U. and Lucke, F.K. 1989 Antibacterial Activity of Lactobacillus sake Isolated from Meat, Applied and Environmental Microbiology, 55 (8), 1901-1906.
[15] Barefoot, S.F. and Klaenhammer, T.R. 1984 Detection and Activity of Lactacin B, a Bacteriocin Produced by Lactobacillus acidophilus, Applied and Environmental Microbiology, 45, 1808-1815.
[16] Lane, D. J. 1991 16S/23S rRNA sequencing. In: Stackebrandt, E. and Goodfellow, M., Ed. Nucleic Acid Techniques in Bacterial Systematics. NewYork, Wiley, pp. 115-175.
[17] www. https://www.ncbi.nlm.nih.gov/
[18] Farber, J.M. and Peterkin, P.I. 1991 Listeria monocytogenes, a Food-borne Pathogen, Microbiology and Molecular Biology Reviews, 55(3), 476-511.
[19] Neysens, P. and De Vuyst, L. 2005 Carbon Dioxide Stimulates the Production of Amylovorin L by Lactobacillus amylovorus DCE 471, While Enhanced Aeration Causes Biphasic Kinetics of Growth and Bacteriocin Production, International Journal of Food Microbiology, 105, 191- 202
[20] Rodriguez, E., Arque, J.L., Rodriguez, R., Nunez, M. and Medina M. 2003 Reuterin Production by Lactobacilli Isolated from Pig Faeces and Evaluation of Probiotic Traits, Letters in Applied Microbiology, 37(3), 259-263.
[21] Ganzle, M.G., Holtzel, A., Walter, J., Jung, G. and Hammes, W. 2000 Characterization of Reutericyclin Produced by Lactobacillus reuteri LTH2584, Applied and Environmental Microbiology, 66(10), 4325-4333.
[22] Garcia-Galaz, A., Perez-Morales, R., Diaz-Cinco, M. and Acedo-Felix, M. 2004 Resistance of Enterococcus Strains Isolated from Pigs to Gastrointestinal Tract and Antagonistic Effect Against Escherichia coli K88, Revista Latinoamericana de Microbiologia, 46(1-2), 5-11.
[23] Yin, Q. and Zheng, Q. 2005 Isolation and Identification of the Dominant Lactobacillus in Gut and Faeces of Pigs using Carbohydrate Fermentation and 16S rDNA Analysis, Journal of Bioscience and Bioengineering, 99(1), 68-71.
[24] Kullen, M.J., Sanozky-Dawes, R.B., Crowell, D.C. and Klaenhammer, T.R. 2000 Use of the DNA Sequence of Variable Regions of the 16S rRNA Gene for Rapid and Accurate Identification of Bacteria in the Lactobacillus acidophilus Complex, Journal of Applied Microbiology, 89, 511-516.
[25] Gueimonde, M., Delgado, S., Mayo, B., Ruas-Madiedo, P., Margolles, A. and de los Reyes-Gavilan, C.G. 2004 Viability and Diversity of Probiotic Lactobacillus and Bifidobacterium Populations Included in Commercial Fermented Milks, Food Resarch International, 37,839-850.
[26] Konstantinov, S.R., Favier, C.F., Zhu, W.Y, Williams, B.A., Klub, J., Souferant, W.B., de Vos, W.M., Akkermans, A.D.L. and Smidt, H. 2004 Microbial Diversity Studies of the Porcine Gastrointestinal Ecosystem during Weaning Transition, Animal Research, 53, 317-324.
[27] Du Toit, M., Dicks, L.M.T. and Holzapfe, W.H. 2003 Identification of Heterofermentative Lactobacilli Isolated from Pig Faeces by Numerical Analysis of Total Soluble Cell Protein Patterns and RAPD-PCR, Letters in Applied Microbiology, 37, 12-16.
[28] Al Jassim, R.A.M. 2003 Lactobacillus ruminis is a Predominant Lactic Acid Producing Bacterium in the Caecum and Rectum of the Pig, Letters in Applied Microbiology, 37(3), 213-217.
[29] Pederson, C. and Roos, S. 2004 Lactobacillus saerimneri sp. nov., Isolated from Pig Faeces, International Journal of Systematic and Evolutionary Microbiology, 54, 1365-1368.
[30] Konstantinov, S.K., Poznanski, E., Fuentes, S. Akkermans, A.D. L., Smidt, H. and de Vos, W.M. 2006 Lactobacillus sobrius sp. nov., Abundant in the Intestine of Weaning Piglets,International Journal of Systematic and Evolutionary Microbiology, 56, 29-32.
[31] Roos, S., Karner, F., Axelsson, L. and Jonsson, H. 2006 Lactobacillus mucosae sp. nov., a New Species with In Vitro Mucus-binding Activity Isolated from Pig Intestine, International Journal of Systematic and Evolutionary Microbiology, 50, 251-258.