Molecular characterization and 16S rRNA sequence analysis of probiotic lactobacillus acidophilus isolated from indigenous Dahi (Yoghurt)
Paper Details
Molecular characterization and 16S rRNA sequence analysis of probiotic lactobacillus acidophilus isolated from indigenous Dahi (Yoghurt)
Abstract
Indigenous yoghurt is being the most popular fermented milk product in Pakistan, produced by heterogenous lactic acid bacteria (LAB). Nowadays LAB has drawn particular interest in food and nutrition science because of functional and probiotic attributes. Among various genera of LAB, Lactobacillus acidophilus is considered to be a potential candidate. Present study was designed by keeping in view the probiotic attributes of L. acidophilus. Isolation was conducted on MRS media supplemented with 0.7% bile salts to screen out bile tolerant isolates. Out of sixty lactobacilli, eighteen were identified as L. acidophilus by performing gram staining, catalase, carbohydrate fermentation test and growth at different temperatures. Only six had shown tolerance of pH 2 up to 50% and only three had shown a wide range of antimicrobial activity against tested organisms. S. epidermis found to be more sensitive with a maximum zone of inhibition (18 mm) and A. baumanii and S. aureus were least sensitive with a smallest zone of inhibition (11.5 mm and 12 mm respectively). 16S rRNA gene amplification, sequencing and phylogenetic tree construction of the obtained sequences with the most closely related lactobacillus spp. was performed. Sequences are available in GeneBank and NCBI with the accession numbers KU877440 (WFA1), KU877441 (WFA2) and KU877442 (WFA3). Presence of probiotic L. acidophilus in locally fermented product (dahi) has proved its potential along with bile salts supplementation in MRS media is helpful in initial screening of probiotic isolates.
Boukhemis M, Djeghri-Hocine B, Tahar A, Amrane A. 2009. Phenotypic characterization of Lactobacillus strains isolated from different biotopes. African Journal of Biotechnology 8(19), 5011-5020. http://dx.doi.org/10.5897/AJB2009.000-9439
Buchanan RE, Gibbons NE. 1974. Bergey’s Manual of Determinative Bacteriology.8th Ed., Williams and Wilkins, Baltimore. p. 1268. http://dx.doi.org/10.1111/j.15507408.1975.tb00935.x
Catanzaro J, Green L. 1997. Microbial ecology and probiotics in human medicine (Part II). Alternative Medical Review 2(4), 296-305.
Clarridge JE. 2004. Impact of 16S rRNA Gene Sequence Analysis for Identification of Bacteria on Clinical Microbiology and Infectious Diseases. Clinical Microbiological Review 17(4), 840-862. http;//dx.doi.org/ 10.1128/CMR.17.4.840862.2004
Donelli G, Vuotto C, Mastromarino P. 2013. Phenotyping and genotyping are both essential to identify and classify a probiotic microorganism. Microbial Ecology in Health and Disease 24, 20105. http://dx.doi.org/10.3402/mehd.v24i0.20105
Goderska K, Czarnecki Z. 2007. Characterization of selected strains from Lactobacillus acidophilus and Bifidobacterium bifidum. African Journal of microbiological Research 1, 65-78.
Harmsen D, Karch H. 2004. 16S rDNA for diagnosing pathogens: a living tree. American Society for Microbiology journal 70, 19-24.
Hassanzadazar H, Ehsani A, Mardani K, Hesari J. 2012. Investigation of antibacterial, acid and bile tolerance properties of lactobacilli isolated from Koozeh cheese. Veterinary Research Forum 3(3), 181-185.
Hoque MZ, Akter F, Hossain KM, Rahman MSM, Billah MM, Islam KMD. 2010. Isolation, identification and analysis of probiotic properties of Lactobacillus spp. from selective regional yoghurts. World Journal of Dairy and Food Science 5, 39-46.
Kaushik JK, Kumar A, Duary RK, Mohanty AK, Grover S, Batish VK. 2009. Functional and Probiotic Attributes of an Indigenous Isolate of Lactobacillus plantarum. Public Library of Science one 4(12), 80-99. http://dx.doi.org/10.1371/journal.pone.0008099.
Kim TW, Song SH, Kim HY. 2005. Distribution of dominant bifido bacteria in the intestinal microflora of Korean adults and seniors, identified by SDS-PAGE of whole cell proteins and 16S rDNA sequence analysis. Journal of Microbiology and Biotechnology 15, 388-394.
Kusmiyati N, Hidayat N, Fatchiyah F. 2014. Partial Analysis 16S rRNA Gene in Lactobacillus spp. from Natural Fermented Milk. Cukurova Medical Journal 39(1), 99-104.
Liong MT, Shah NP. 2005. Acid and bile tolerance and cholesterol removal ability of lactobacilli strain. Journal of Dairy Science 88, 55-66. http://dx.doi.org/10.3168/jds.S00220302(05)72662-X
Liu X, Liu W, Zhang Q, Tian F, Wang G, Zhang H, Chen W. 2013. Screening of lactobacilli with antagonistic activity against enteroinvasive Escherichia coli. Food Control, 3, 563-568.
Ljungh A, Wadström T. 2006. Lactic acid bacteria as probiotics. Current Issues in Intestinal Microbiology 7(2), 73-89.
Mandal S, Puniya AK, Singh K. 2006. Effect of alginate concentration on survival of encapsulated Lactobacillus casei NCDC-298. International Dairy Journal 16, 1190-1195. http://dx.doi.org/10.1016/j.idairyj.2005.10.005
Mishra V, Prasad DN. 2005. Application of in vitro methods for selection of Lactobacillus casei strains as potential probiotics. International Journal of Food Microbiology 103, 109-115. http://dx.doi.org/10.1016/j.ijfoodmicro.2004.10.047
Ohmomo S, Murata S, Katayama N, Nitisinprasart S, Kobayashi M, Nakajima T, Yajima M, Nakanishi K. 2000. Purification and some characteristics of enterocin ON-157, a bacteriocin produced by Enterococcus faecium NIAI 157. Journal of Applied Microbiology 88, 81-89. http://dx.doi.org/10.1046/j.1365-2672.2000.00866.x
Palys T, Nakamura LK, Cohan FM. 1997. Discovery and classification of ecological diversity in the bacterial world: the role of DNA sequence data. International Journal of Systematic Bacteriology 47, 1145-1156. http://dx.doi.org/10.1099/00207713-47-4-1145
Plourde-Owobi L, Seguin D, Baudin MA, Moste C, Rokbi B. 2005. Molecular characterization of Clostridium tetani strains by pulsed-field gel electrophoresis and colony PCR. Applied and Environmental Microbiology 71(9), 5604-5606. http://dx.doi.org/10.1128/AEM.71.9.56045606.2005
Prasad J, Gill H, Smart J. 1998. Selection and characterization of Lactobacillus and Bifidobacterium strains for use as probiotics. International Dairy Journal 8, 993-1002. http://dx.doi.org/10.1016/s0958-6946(99)00024-2
Pyar H, Peh KK. 2014. Characterization and identification of Lactobacillus acidophilus using biolog rapid identification system. International Journal of Pharmacy and Pharmaceutical Sciences 6(1), 189-193.
Rahayu TH, Gandjar I, Riani E, Djunaidah IS, Sjamsuridzal A. 2009. Identification and Phylogenetic Analysis of Bacterial Isolates from Litopenaeus vannamei Shrimp Culture System and Gut Environment Based on 16SrRNA Gene Sequence Data. Microbiology Indonesia 3(2), 56-60.
J, Gunasekaran P. 2011. Microbial phylogeny and diversity: Small subunit ribosomal RNA sequence analysis and beyond. Microbiological Research 166(2), 99-110. http://dx.doi.org/10.1016/j.micres.2010.02.003.
Raquib M, Trishna B, Choudhary RK, Rahaman H, Borpuzari T. 2003. Isolation and characterization of lactobacilli isolated from market sample of sour indigenous yoghurt. Indian Veterinary Journal 80, 791-794.
Reid G, Sanders ME, Gaskins HR, Gibson GR, Mercenier A, Rastall R, Roberfroid M, Rowland I, Cherbut C, Klaenhammer TR. 2003. New scientific paradigms for probiotics and prebiotics. Journal of Clinical Gastroenterology 37, 105-118.
Singhal K, Joshi H, Chaudhary BL. 2010. Bile and acid tolerance ability of probiotic Lactobacillus Strains. Journal of global pharma Technology 2, 17- 25.
Soomro TH, Masud T. 2012. Probiotic characteristics of Lactobacillus spp. isolated from fermented milk product indigenous yoghurt. Food Science and Technology Research 18, 91-98.
Tortoli E. 2003. Impact of genotypic studies on mycobacterial taxonomy: the new mycobacteria of the 1990s. Clinical Microbiology Reviews 16, 319-354. http://dx.doi.org/10.1128/CMR.16.2.319354.2003
Tringe SG. Hugenholtz P. 2008. A renaissance for the pioneering 16S rRNA gene. Current Opinion in Microbiology 11, 442-446. http://dx.doi.org/10.1016/j.mib.2008.09.011
Wajiha Farid, Tariq Masud, Asma Sohail, Saqlain Naqvi, Mudassir Ali Qazalbash, 2016. Molecular characterization and 16S rRNA sequence analysis of probiotic lactobacillus acidophilus isolated from indigenous Dahi (Yoghurt). Int. J. Biosci., 9(5), 19-27.
Copyright © 2016 by the Authors. This article is an open access article and distributed under the terms and conditions of the Creative Commons Attribution 4.0 (CC BY 4.0) license.