SCREENING OF POTENTIAL PROBIOTIC LACTOBACILLUS STRAINS ISOLATED FROM FERMENTED FOODS, FRUITS AND OF HUMAN ORIGIN
Abstract
Objective: To screen and characterize Lactobacillus strains having potential probiotic properties from fermented foods and fruits, and to investigate cell surface characteristics and antimicrobial activity against food-borne and gastrointestinal tract (GIT) pathogens.
Methods and Results: In the present study, twenty five Lactobacillus strains were isolated from fruits, fermented foods and human origin on De Man Rogosa and Sharpe (MRS) agar plates and identified on the basis of their phenotypic characteristics and 16s rDNA sequencing. Isolated Lactobacillus strains tolerated inhibitory substances like bile (upto 4%), NaCl (upto 8%), phenol (upto 0.6%) and low pH (2.0). Six of the isolates exhibited inhibitory activity against food-borne and GIT pathogens by well diffusion method. The isolates appeared to possess hydrophilic cell surface as determined by microbial adhesion to n-hexadecane. Isolates auto-aggregated and also co-aggragated with pathogens tested. The isolates were not susceptible to antibiotics like vancomycin, tobramycin, kanamycin and norfloxacin while were highly susceptible to chloramphenicol, clindamycin, erythromycin, oleandomycin and penicillin G. None of the isolates showed haemolytic activity.
Conclusion: Six strains (Lactobacillus paracasei HML1, L. paracasei CR7, L. plantarum BRMV1, L. plantarum F1 and L. plantarum OC6) showed potential probiotic properties as well as strong in vitro antibacterial activity against various food-borne and gastrointestinal tract (GIT) pathogens.Â
Keywords: Antimicrobial activity, Lactobacillus, Acid tolerance, Bile tolerance, Hydrophobicity, Probiotics
Â
Downloads
References
Guasch-Jané M, Andrés-Lacueva C, Jáuregui O, Lamuela- Raventós R. First evidence of white wine in ancient Egypt from Tutankhamun’s tomb. J Archaeol Sci 2005; 33: 1075-1080.
Azcarate-Peril AM, Raya RR. In: J.F.T. Spencer and A.L.R.D. Spencer, editors. New Jersey: Humana Press Inc; 2001. p: 135.
Tannock, G. A special fondness for lactobacilli. Appl Environ Microbiol 2004; 70: 3189-3194.
Gardiner G, Heinemann C, Baroja M, Bruce A, Beuerman D, Madrenas J, Reid G. Oral administration of the probiotic combination Lactobacillus rhamnosus GR-1 and L. fermentum RC-14 for human intestinal applications. Int Dairy J 2002; 12:191-196.
Kleerebezemab M, Hols P, Hugenholtz J. Lactic acid bacteria as a cell factory: rerouting of carbon metabolism in Lactococcus lactis by metabolic engineering. Enzy Microbial Technol 2002; 26:840-848.
Hasan NA, Frank JF. In: Marth EH and Steele JL, editors. Applied Dairy Microbiology 2nd ed. New York: Marcel Dekker, Inc; 2001. p. 152-155.
Vernoux JP, Coeuret V, Dubernet S, Bernardeau M, Gueguen M. Isolation, characterization and identification of lactobacilli focusing mainly on cheeses and other dairy products. INRA EDP Sci 2003; 83:269-306.
Fuller R. Probiotics in man and animals. J Appl Bacteriol 1989; 66: 365-378.
Weinberg Z, Shatz O, Chen Y, Yosef E, Nikbahat M, Ben-Ghedalia D, Miron J. Effect of lactic acid bacteria inoculants on in vitro digestibility of wheat and corn silages. J Dairy Sci 2007; 90: 4754-4762.
Abdelbasset M, K Djamila. Antimicrobial activity of autochthonous lactic acid bacteria isolated from Algerian traditional fermented milk Raïbâ€. Afr J. Biotechnol 2008; 7: 2908-2914.
Temmerman R, Pot B, Huys G, Swings J. Identification and antibiotic susceptibility of bacterial isolates from probiotic products. Int J Food Microbiol 2002; 81:1-10.
Kalliomäki M, Salminen S, ArvilommiH, Kero P, Koskinen P, Isolauri E. Probiotics in primary prevention of atopic disease: a randomised placebo-controlled trial. The Lancet 2001; 357:1076-1079.
Catanzaro J, Green L. Microbial ecology and probiotics in human medicine (Part II). Altern Med Rev1997; 2:296-305.
Walker DK, Gilliland SE. Relationship among bile tolerance, bile salt deconjugation and assimilation of cholesterol by Lactobacillus acidophilus. J Dairy Sci 1983; 76:956-961.
Kilara A. Influence of in vitro gastric digestion on survival of some lactic cultures. Milchwissenschaft 1982; 37:129-132.
Graciela F, V. De, MarÃa PT. In: J.F.T. Spencer and A.L.R.D. Spencer, editors. Food microbiology protocols. New Jersey: Humana Press Inc. 2001. p.173-181.
Pedersen K, Tannock GW. Colonization of the porcine gastrointestinal tract by lactobacilli. Appl Env Microbiol 1989; 55:279-283.
Bibiloni R, Perez PF, Garrote GL, Disalvo EA, De Antoni GL. Surface characterization and adhesive properties of bifidobacteria. Methods Enzymol 2001; 336:411–27.
Jankovic I, Ventura M, Meylan V, Rouvet M, Elli M, Zink R. Contribution of aggregation-promoting factor to maintenance of cell shape in Lactobacillus gasseri 4B2. J Bacteriol 2003; 185:88-96.
Reid G, McGroarty JA, Angotti R, Cook RL. Lactobacillus inhibitor production against Escherichia coli and coaggregation ability with uropathogens. Can J Microbiol 1988; 34:344-351.
Wadstro¨m T, Andersson K, Sydow M, Axelsson L, Lindgren S, Gullmar B. Surface properties of lactobacilli isolated from the small intestine of pigs. Journal of Applied Bacteriology 1987; 62:513–520.
Del Re B, Sgorbati B, Miglioli M, Palenzona, D. Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Let Applied Microbiol 2000; 31:438-442.
Schneitz C, Nuotio L, Lounatma K. Adhesion of Lactobacillus acidophilus to avian intestinal epithelial cells mediated by the crystalline bacterial cell surface layer (S-layer). J App Bacteriol 1993; 74:290-294.
Mukai T, Arihara K. Presence of intestinal lectin-binding glycoproteins on the cell surface of Lactobacillus acidophilus. Biosci Biotechnol Biochem 1994; 58:1851-1854.
Gill HS, Guarner F. Probiotics and human health: a clinical perspective. Postgrad Med J 2004; 80: 516-526.
Dunne C, O’Mahony L, Murphy L, Thonton G, Morrissey D, O’Halloran S et al. In vitro selection criteria for probiotic bacteria of human origin: correlation with in vivo findings. Am J Clin Nutr 2001; 73:386S-392S.
Van der kaaij, Desiere H, Mollet B, Germond J. L-alanine auxtrophy of Lactobacillus johnsonii as demonstrated by physiological, genomic and gene complementation approaches. Appl Env Microbiol 2004; 70:1869-1873.
Ogunshe AAO, Omotoso MA, Adeyeye JA. In vitro antimicrobial characteristics of bacteriocins producing Lactobacillus strains from Nigrian indigenous fermented foods. Afri J Biotechnol 2007; 6:445-453.
De Man J, Rogosa M, Sharpe M. A medium for the cultivation oflactobacilli.J.Appl. Bacteriol 1960; 3:13-135.
Kebede A, Viljoen B, Gadaga T, Narvhus J, Lourens- HattinghA.The effect of container type on the growth of yeast and lactic acidbacteria during production of SethemiSouth African spontaneouslyfermented milk. Food Res Int 2007; 40:33-35.
Kandler O, N Weiss.Bergey'sManual of Systematic BacteriologyIn: P. H. A.Sneath, N. S. Mair, M. E. Sharpe, J. G. Holt editors. Baltimore: Williams and Wilkins; 1986 p.1209-1234.
SharpeME.Identification of the lacticacidbacteria. In: Skinner, FA and Lovelock, EditorsIdentification methods formicrobiologists. London: Academic Press; 1979 p. 233-259.
Ambalam PS, Prajapati J B, Dave J M, Nair B, Ljungh A, VyasBRM.Isolation and characterization of antimicrobial protein produced by a potential probiotic strain of human Lactobaciilusrhamnosus231 and its effect on selected human pathogens and food spoilage organisms MicrobEcol Health Disease 2009; 21:211-220.
Turner S, Pryer KM, Miao VPW, Palmer JD. Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J EukaryotMicrobiol 1999; 46:327–338.
Liong MT, Shah NP.Acid and bile tolerance and the cholesterol removal ability of Bifidobacteriastrains.BiosciMicroflora 2005;24:1-10.
Crow VL, Gopal PK. Cell surface differences of lactococcal strains. International Dairy Journal 1995; 5:45-68.
Bellon-Fontaine, Rault MN, van Oss CJ. Microbial adhesion to solvents: a novel method to determine the electrondonor/electron-acceptor or Lewis acid-base properties of microbialcells. Colloids and Surfaces 1996; 7, 47-53.
Lindahl M, Faris A, Wadstrom T, Hjerten S. A new test based on ‘salting out’ to measure relative surface hydrophobicity of bacterial cells. BiochimBiophysActa1981; 77:471-476.
Todorov SD, Dicks LMT. Evaluation of lactic acid bacteria from kefir, molasses and olive brine as possible probiotics based on physiological properties. Ann Microbiol 2008; 58:661-670.
Flemming HP, Etchells JL, Costilow RL. Microbial inhibition by an isolate of pediococcus from cucumber brines. Applied Microbiol 1985;30:1040-1042.
Mourad K, Eddine KN. In vitro preselection criteria for probiotic Lactobacillus plantarumstrains of fermented olives origin. Int. J. Probiotics Prebiotics 2006; 1:27-32.
Reid G.The importance of guidelines in the development and application of Probiotics.Curr. Pharm. Des 2005;11:11-16.
Dunne C, O’Mahony L, Murphy L, Thornton G, Morrisey D, O’Halloran S et al. In vitro selection criteria for probiotic bacteria of human origin: Correlation with in vivo findings. Am J Clin Nutr 2001; 73:386-392.
Conway PL, Gorbach SL, Goldin BR. Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells. J Dairy Sci 1987; 70:1-12.
Minelli EB, Benini A, Marzotto M, Sbarbati A, Ruzzenente O, Ferrario R et al. Assessment of novel probiotic Lactobacillus casei strains for the production of functional dairy foods. Int Dairy J 2004; 14:723-736.
Noriega L, Gueimonde M, Sanchez B, Margolles A, de los Reyes-Gavilan C. Effect of the adaptation to high bile salts concentrations on glycosidic activity, survival at low pH and cross-resistance to bile salts in Bifidobacterium. Int J Food Microbiol 2004; 94:79-86.
Elizete DFRP, RS Carlos. Biochemical characterization and identification of probiotic Lactobacillus for swine. B.CEPPA, Curitiba 2005; 23:299-310.
Sathyabama S, Vijayabharathi R, Bruntha devi P, Ranjithkumar M, Priyadarisini V. Screening for probiotic properties of strains isolated from feces of various human groups. J Microbiol 2012; 50:603-612.
Rojas M, Ascencio F, Conway PL. Purification and characterization of a surface protein from Lactobacillus fermentum104R that binds to porcine small intestinal mucus and gastric mucin. Appl. Environ. Microbiol 2002; 68:2330-2336.
Ross S, Jonsson H. A high-molecular mass cell surface protein from Lactobacillus reuteri1063 adheres to mucus components. Microbiol 2002;148:433-442.
Kiely LJ, Olson NF. The physicochemical surface characteristics of Lactobacillus casei. Food Microbiol 2000; 17:277-291.
Todorov SD, Dicks LMT (2008). Evaluation of lactic acid bacteria from kefir, molasses and olive brine as possible probiotics based on physiological properties. Ann. Microbiol. 58:661-670.
Kleerebezem M, Boekhorst J, van Kranenburg R, Molenaar D, Kuipers OP, Leer Ret al. Competitive genome sequence of Lactobacillus plantarumWCFS1. Proc Natl Acad Sci 2003; 100:1990-1995.
Reid G, McGroarty JA, Angotti R, Cook, RL. Lactobacillus inhibitor production against Escherichia coli and coaggregation ability with uropathogens. Can J of Microbiol 1988; 34:344-351.
Spencer, RJ, Chesson A. The effect of Lactobacillus spp. on the attachment of enterotoxigenic Escherichia coli to isolated porcine enterocytes. J App Bacteriol 1994; 77:215-220.
Pithva SP, Shekh S, Dave JM, Vyas BRM. Probiotic attributes of autochthonous Lactobacillus rhamnosus strains of human origin. Appl Biochem Biotechnol 2014; DOI 10.1007/s12010-014-0839-9.
El-Naggar MYM. Comparative study of probiotic cultures to control the growth of Escherichia coli O157:H7 and Salmonella typhimurium. Asian Network for Scientific Information Biotechnol 2004; 3:173-180.
Published
How to Cite
Issue
Section
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.