SCREENING, CHARACTERIZATION, AND IN VITRO EVALUATION OF PROBIOTIC PROPERTIES OF LACTOBACILLUS STRAINS
Â Objective: The aim of the present investigation was to isolate and identify Lactobacillus strains from dairy and cattle dung samples. Potent isolates were selected for screening by antimicrobial activity; selected lactobacilli were further tested for probiotic properties and adhesive attributes.
Methods: Lactobacilli were isolated aseptically on specific de man, rogosa and sharpe medium from dairy and cattle dung samples. Isolates were identified by Gram-staining, motility, catalase, endospore, and carbohydrate fermentation tests. Further, the isolates were screened for antimicrobial activity by disk diffusion assay, and potent lactobacilli were observed for probiotic properties: Acid and bile salt tolerance, gelatinase activity, and autolytic activity. For analyzing the adhesive attributes, isolates were observed for autoaggregation, coaggregation and microbial adhesion to solvents assay.
Results: About 12 Lactobacillus strains among 98 isolates exhibited maximum antimicrobial activity were further selected for identifying their probiotic and adhesive attributes. Among 12 selected isolates, cell-free supernatant (CFS) of buffalo milk BM10 and goat milk GM10 showed excellent antimicrobial activity, 20.34Â±0.02 mm against Staphylococcus aureus and 18.65Â±0.11 mm against Escherichia coli. Isolates showed survival at pH 2 and 3 and can tolerate 0.2-0.3% bile salt concentrations. The GM5 showed maximum autoaggregation (67.04Â±0.61%) and minimum coaggregation (11.51Â±0.50%) showed by GM3. The BM10 exhibited maximum adherent value 64.84Â±1.41% for n-hexadecane.
Conclusion: The two lactobacilli, BM10 and GM10 identified as Lactobacillus fermentum and Lactobacillus pentosus on the basis of phenotypic and sugar utilization tests. The CFS of both lactobacilli can be used as antimicrobial agent. Both isolates showed significant results of probiotic and adhesive attributes, therefore, can be evaluated for clinical and therapeutic applications.
2. Mishra V, Prasad DN. Application of in vitro methods for selection of Lactobacillus casei strains as potential probiotics. Int J Food Microbiol 2005;103(1):109-15.
3. Pakdaman MN, Udani JK, Molina JP, Shahani M. The effects of the DDS-1 strain of Lactobacillus on symptomatic relief for lactose intolerance - A randomized, double-blind, placebo-controlled, crossover clinical trial. Nutr J 2016;15(1):2-11.
4. Amdekar S, Roy P, Singh V, Kumar A, Singh R, Sharma P. Anti-inflammatory activity of lactobacillus on carrageenan-induced paw edema in male wistar rats. Int J Inflam 2012;2012:752015.
5. Bouhafs L, Moudiloub EN, Exbrayat JM, Lahouel M, Idouiad T. Protective effects of probiotic Lactobacillus plantarum BJ0021 on liver and kidney oxidative stress and apoptosis induced by endosulfan in pregnant rats. Ren Fail 2015;37(8):1370-8.
6. Dilna SV, Surya H, Aswathy RG, Varsha KK, Sakthikumar DN, Pandey A, et al. Characterization of an exopolysaccharide with potential health benefits properties from a probiotic Lactobacillus plantarum RJF4. Food Sci Technol 2015;64:1179-86.
7. Gaon D, Garmendia C, Murrielo NO, Games AD, Cerchio A, Quintas R, et al. Effect of Lactobacillus strains (L. casei and L. acidophilus strains cereal) on bacterial overgrowth-related chronic diarrhea. Medicina 2002;62:159-63.
8. Kaewnopparat S, Dangmanee N, Kaewnopparat N, Srichana T, Chulasiri M, Settharaksa S. In vitro probiotic properties of Lactobacillus fermentum SK5 isolated from vagina of a healthy woman. Anaerobe 2013;22:6-13.
9. Bejar W, Hamden K, Ben Salah R, Chouayekh H. Lactobacillus plantarum TN627 significantly reduces complications of alloxan-induced diabetes in rats. Anaerobe 2013;24:4-11.
10. Liu CF, Tung YT, Wu CL, Lee BH, Hsu WH, Pan TM. Antihypertensive effects of Lactobacillus-fermented milk orally administered to spontaneously hypertensive rats. J Agric Food Chem 2011;59(9):4537-43.
11. Lee HY, Park JH, Seok SH, Baek MW, Kim DJ, Lee KE, et al. Human originated bacteria, Lactobacillus rhamnosus PL60, produce conjugated linoleic acid and show anti-obesity effects in diet-induced obese mice. Biochim Biophys Acta 2006;1761(7):736-44.
12. Narva M, Collin M, Lamberg-Allardt C, KÃ¤rkkÃ¤inen M, Poussa T, Vapaatalo H, et al. Effects of long-term intervention with Lactobacillus helveticus-fermented milk on bone mineral density and bone mineral content in growing rats. Ann Nutr Metab 2004;48(4):228-34.
13. Deegan LH, Cotter PD, Hill C, Ross P. Bacteriocins: Biological tools for bio-preservation and shelf-life extension. Int Dairy J 2006;16:1058-71.
14. Grajek K, Sip A, Foksowicz-Flaczyk J, Dobrowolska A, Wita A. Adhesive and hydrophobic properties of the selected LAB isolated from gastrointestinal tract of farming animals. Acta Biochim Pol 2016;63(2):311-4.
15. Wen LS, Philip K, Ajam N. Purification, characterization and mode of action of plantaricin K25 produced by Lactobacillus plantarum. Food Control 2015;60:430-9.
16. Fong FL, Kirjavainen P, Wong VH, Nezami HE. Immunomodulatory effects of Lactobacillus rhamnosus GG on dendritic cells, macrophages and monocytes from healthy donors. J Funct Foods 2015;13:71-9.
17. Hartemink R, Domenech VR, Rombouts FM. LAMVAB - A new selective medium for the isolation of lactobacilli from faeces. J Microbiol Methods 1997;29(2):77-84.
18. Esayas A, Fekadu B, Amutha S. Isolation and characterization of inhibitory substance producing lactic acid bacteria from Ergo, Ethiopian traditional fermented milk. Livest Res Rural Dev 2008;20:1-8.
19. Sieladie DV, Zambou NF, Kaktcham PM, Cresci A, Fonteh F. Probiotic properties of Lactobacilli strains isolated from raw cow milk in the western highlands of Cameroon. Innov Rom Food Biotechnol 2011;9:12-28.
20. Dora IA, Glenn RG. Cholesterol assimilation by lactic acid bacteria and bifidobacteria isolated from the human gut. Appl Environ Microbiol 2002;68(9):4689-93.
21. Harrigan WF, McCance ME. Laboratory Methods in Food and Dairy Microbiology. London: Academic Press; 1990.
22. Basson A, Flemming LA, Chenia HY. Evaluation of adherence, hydrophobicity, aggregation, and biofilm development of Flavobacterium johnsoniae-like isolates. Microb Ecol 2008;55(1):1-14.
23. Saran S, Bisht MS, Singh K, Teotia UV. Compare adhesion attributes of two isolates of Lactobacillus acidophilus for assessment of prebiotics, honey and inulin. Int J Sci Res Publ 2012;2(4):1-7.
24. Kos B, Suskovic J, Vukovic S, Simpraga M, Frece J, Matosic S. Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 2003;94:981-7.
25. Mohanty D, Saini MR, Mohapatra S. In vitro study on release of bioactive antimicrobial compounds from dairy products by certain promising probiotic Lactobacillus strains. Int J Pharm Pharm Sci 2017;9:2-6.
26. Singh TP, Kaur G, Malik RK, Schillinger U, Guigas C, Kapila S. Characterization of intestinal Lactobacillus reuteri strains as potential probiotics. Probiotics Antimicrob Proteins 2012;4:47-58.
27. Haung Y, Adams MC. In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionic bacteria. Int J Food Microbiol 2004;91(3):253-60.
28. Mohanty D, Ray P. Evaluation of probiotic and antimicrobial properties of Lactobacillus strains isolated from dairy products. Int J Pharm Pharm Sci 2016;8(11):230-4.
29. Prabhurajeshwar C, Kelmani CR. Development of in vitro methodologies for inhibition of pathogenic bacteria by potential probiotic Lactobacillus sps; An evidence for production of antimicrobial substances. Int J Pharm Pharm Sci 2016;8(12):277-86.
30. Koch S, Eugster-Meier E, Oberson G, Meile L, Lacroix C. Effects of strains and growth conditions on autolytic activity and survival to freezing and lyophilization of Lactobacillus delbrueckii ssp. Lactis isolated from cheese. Int Dairy J 2007;18(2):187-96.
31. GarcÃa-Cayuela T, Korany, AM, Bustos I, de CadiÃ±anos LP, Requena T, PelÃ¡ez C, et al. Adhesion abilities of dairy Lactobacillus plantarum strains showing an aggregation phenotype. Food Res Int 2014;57:44-50.
32. Kruisselbrink A, Bak-Glashouwer MJ, Havenith CE, Thole JE, Janssen R. Recombinant Lactobacillus plantarum inhibits house dust mite-specific T-cell responses. Clin Exp Immunol 2001;126(1):2-8.
33. Gudina EJ, Teixeira JA, Rodrigues LR. Isolation and functional characterization of a biosurfactant produced by Lactobacillus paracasei. Colloids Surf B Biointerfaces 2010;76(1):298-304.
34. Kiely LJ, Olson NF. The physicochemical surface characteristics of Lactobacillus casei. Food Microbiol 2000;17(3):277-91.
35. Del Re B, Sgorbati B, Miglioli M, Palenzola D. Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett Appl Microbiol 2000;31(6):438-42.
36. Nikolic M, Jovcic B, Kojic M, Topisirovic L. Surface properties of Lactobacillus and Leuconostoc isolates from homemade cheeses showing auto-aggregation ability. Eur Food Res Technol 2010;231(6):925-31.
37. Palaniswamy SK, Govindaswamy V. In-vitro probiotic characteristics assessment of feruloyl esterase and glutamate decarboxylase producing Lactobacillus spp. Isolated from traditional fermented millet porridge (kambu koozh). LWT Food Sci Technol 2016;68:208-16.
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