AiM  Vol.7 No.4 , April 2017
Symbiotic, Hypocholesterolemic and Antioxidant Effects of Potential Probiotic Lactobacilli Strains Isolated from Tunisian Camel Milk
Abstract: In the present study, 20 selected Lactobacillus strains already characterized in a previous research for their capability to grow in conditions simulating the intestinal environment, their resistance to antibiotics, their antibacterial activity and their adhesion capability to intestinal human Caco-2 TC7 and HT-29 MTX cell lines, were further investigated to explore more their probiotic properties. Growth behaviour in the presence of prebiotic (fructooligosac-charides (FOS) and lactulose) at a concentration of 2%, cholesterol removal by measuring the residual cholesterol in medium supplemented with cholesterol, ability to deconjugate bile salts using BSH enzyme and antioxidant activity of culture supernatant of Lactobacillus strains by ABTS·+ and DPPH methods were analyzed. All probiotic strains demonstrated important prebiotic assimilation (P > 0.05) even with OD600 > 3 after 30 h of contact, cholesterol removal ability with maximum percentage of 57% after 24 h of contact and they were found to liberate significantly (P < 0.05) more cholic acid with maximum of 0.40 mM of sodium glycocholate, 0.33 mM of sodium taurocholate and 0.41 mM of their mixte and scavenge both radicals with 52% and 2.19% of ABTS·+ and DPPH respectively. This study confirmed the suitability of these probiotic strains for application in functional food formulations especially where cholesterol reduction and antioxidant activity in food are needed to assess possible in vivo human health benefits.
Cite this paper: Mahmoudi, I. , Moussa, O. and Hassouna, M. (2017) Symbiotic, Hypocholesterolemic and Antioxidant Effects of Potential Probiotic Lactobacilli Strains Isolated from Tunisian Camel Milk. Advances in Microbiology, 7, 328-342. doi: 10.4236/aim.2017.74027.

[1]   Haghshenas, B., Nami, Y., Abdullah, N., Radiah, D., Rosli, R. and Khosroushahi, A.Y. (2015) Anticancer Impacts of Potentially Probiotics Acetic Acid Bacteria Isolated from Traditional Dairy Microbiota. LWT Food Science and Technology, 60, 690-697.

[2]   Guarner, F., Khan, A. G., Garisch, J., Eliakim, R., Gangl, A., Thomson, A., Krabshuis, J. and Lemair, T. (2008) Probiotics and Prebiotics. World Gastroenterology Organisation Practice Guideline.

[3]   Casiraghi, M.C., Canzi, E., Zanchi, R., Donati, E. and Villa, L. (2007) Effects of a Synbiotic Milk Product on Human Intestinal Ecosystem. Journal of Applied Microbiology, 103, 499-506.

[4]   Huebner, J., Wehling, R.L. and Hutkins, R.W. (2007) Functional Activity of Commercial Prebiotics. International Dairy Journal, 17, 770-775.

[5]   Shah, N.P. (2001) Functional Foods, Probiotics and Prebiotics. Food Technology, 55, 46-53.

[6]   Isolauri, E. (2004) The Role of Probiotics in Paediatrics. Current Paediatrics, 14, 104-109.

[7]   Roberfroid, M.B. (2000) Prebiotics and Probiotics: Are They Functional Foods. American Journal of Clinical Nutrition, 71, 1682-1687.

[8]   Park, S.F. and Kroll, R.G. (1993) Expression of Lysteriolysin and Phosphatidylinositol-Specific Phospholipase C Is Repressed by the Plant-Derived Molecule Cellobiose in Listeria monocytogenes. Molecular Microbiology, 8, 653-661.

[9]   Fric, P. (2007) Probiotics and Prebiotics—Renaissance of a Therapeutic Principle. European Journal of Medicinal Chemistry, 2, 237-270.

[10]   Wang, J., Zhang, H., Chen, X., Chen, Y., Menghebilige and Bao, Q. (2012) Selection of Potential Probiotic Lactobacilli for Cholesterol-Lowering Properties and Their Effect on Cholesterol Metabolism in Rats Fed a High-Lipid Diet. Journal of Dairy Science, 95, 1645-1654.

[11]   Shahidi, F. (2012) Nutraceuticals, Functional Foods and Dietary Supplements in Health and Disease. Journal of Food and Drug Analysis, 22, 226-230.

[12]   Mann, G.V. and Spoerry, A. (1974) Studies of a Surfactant and Cholesteremia in the Maasai. American Journal of Clinical Nutrition, 27, 464-469.

[13]   Park, S.C., Hwang, M.H., Kim, Y.H., Kim, J.C., Song, J.C., Lee, K.W., Jeong, K.S., Rhee, M.H., Kim, K.S. and Kim, T.W. (2006) Comparison of pH and Bile Resistance of Lactobacillus acidophilus Strains Isolated from Rat, Pig, Chicken, and Human Sources. World Journal of Microbiology and Biotechnology, 22, 35-37.

[14]   Miremadi, F., Ayyash, M., Sherkat, F. and Stojanovska, L. (2014) Cholesterol Reduction Mechanisms and Fatty Acid Composition of Cellular Membranes of Probiotic Lactobacilli and Bifidobacteria. Journal of Functional Foods, 9, 295-305.

[15]   Wang, Y.P., Xu, N., Xi, A.D., Ahmed, Z., Zhang, B. and Bai, X.J. (2009) Effects of Lactobacillus plantarum MA2 Isolated from Tibet Kefir on Lipid Metabolism and Intestinal Microflora of Rats Fed on High-Cholesterol Diet. Applied in Microbiology and Biotechnology, 84, 341-347.

[16]   Ataie-Jafari, A., Larijani, B., Majd, H.A. and Tahbaz, F. (2009) Cholesterol-Lowering Effect of Probiotic Yogurt in Comparison with Ordinary Yogurt in Mildly to Moderately Hypercholesterolemic Subjects. Annals of Nutrition and Metabolism, 54, 22-27.

[17]   Lye, H.S., Rusul, G. and Liong, M.T. (2010) Removal of Cholesterol by Lactobacilli via Incorporation and Conversion to Coprostanol. Journal of Dairy Science, 93, 1383-1392.

[18]   Zhang, F., Hang, X., Fan, X., Li, G. and Yang, H. (2007) Selection and Optimization Procedure of Synbiotic for Cholesterol Removal. Anaerobe, 13, 185-192.

[19]   Ha, C.G., Cho, J.K., Lee, C.H., Chai, Y.G., Ha, Y.A. and Shin, S.H. (2006) Cholesterol Lowering Effect of Lactobacillus plantarum Isolated from Human Feces. Journal of Microbiology and Biotechnology, 16, 1201-1209.

[20]   Corzo, G. and Gilliland, S.E. (1999) Bile Salt Hydrolase Activity of Three Strains of Lactobacillus acidophilus. Journal of Dairy Science, 82, 472-480.

[21]   Liong, M.T. and Shah, N.P. (2005) Acid and Bile Tolerance and Cholesterol Removal Ability of Lactobacilli Strains. Journal of Dairy Science, 88, 55-66.

[22]   Walker, D.K. and Gilliland, S.E. (1993) Relationship among Bile Tolerance, Bile Salt Deconjugation, and Assimilation of Cholesterol by Lactobacillus acidophilus. Journal of Dairy Science, 76, 956-961.

[23]   Dambekodi, P.C. and Gilliland, S.E. (1998) Incorporation of Cholesterol into the Cellular Membrane of Bifidobacterium longum. Journal of Dairy Science, 81, 1818-1824.

[24]   Pieniz, S., Andreazza, R., Anghinoni, T., Camargo, F. and Brandelli, A. (2014) Probiotic Potential, Antimicrobial and Antioxidant Activities of Enterococcus durans Strain LAB18s. Food Control, 37, 251-256.

[25]   Mahmoudi, I., Ben Moussa, O., Khaldi, T.E., Kebouchi, M., Soligot, C., Le Roux, Y. and Hassouna, M. (2016) Functional in Vitro Screening of Lactobacillus Strains Isolated from Tunisian Camel Raw Milk toward Their Selection as Probiotic. Small Ruminant Research, 137, 91-98.

[26]   Pennacchia, C., Vaughan, E.E. and Villani, F. (2006) Potential Probiotic Lactobacillus Strains from Fermented Sausages: Further Investigations on Their Probiotic Properties. Meat Science, 73, 90-101.

[27]   Pilch, P., Radziszewski, P. and Maciukiewicz, P. (2012) Prostate Cancer Dependance upon Cholesterol, Statins and Diet. Rak Gruczolu Krokowego a Cholesterol, Statyny i Dieta, 65, 31-37.

[28]   Rossini, K., Noreña, C.P.Z., Olivera, F.C. and Brandelli, A. (2009) Casein Peptides with Inhibitory Activity on Lipid Oxidation in Beef Homogenates and Mechanically Deboned Poultry Meat. LWT Food Science and Technology, 42, 862-867.

[29]   Brand-Williams, W., Cuvelier, M.E. and Berset, C. (1995) Use of Free Radical Method to Evaluate Antioxidant Activity. LWT Food Science and Technology, 2, 25-30.

[30]   Kolida, S., Tuohy, K. and Gibson, G.R. (2002) Prebiotic Effects of Inulin and Oligosaccharydes. British Journal of Nutrition, 87, S193-S197.

[31]   Gibson, G.R., Beatty, E.R., Wang, X. and Cummings, J.H. (1995) Selective Stimulation of Bifidobacteria in the Human Colon by Oligofructose and Inulin. Gastroenterology, 108, 975-982.

[32]   Delzenne, N.M. (2003) Oligosaccharides: State of the Art. Proceedings of the Nutrition Society, 62, 177-182.

[33]   Saarela, M., Hallamaa, K., Mattila-Sandholm, T., and Matto, J. (2003) The Effect of Lactose Derivates Lactulose, Lactitol and Lactobionic Acid on the Functional and Technological Properties of Potentially Probiotic Lactobacillus Strains. International Dairy Journal, 13, 291-302.

[34]   Kaplan, H. and Hutkins, R.W. (2000) Fermentation of Fructooligosaccharides by Lactic Acid Bacteria and Bifidobacteria. Applied Environmental Microbiology, 66, 2682-2684.

[35]   Palframan, R.J., Gibson, G.R. and Rastall, R.A. (2002) Effect of pH and Dose on the Growth of Gut Bacteria on Prebiotic Carbohydrates in Vitro. Anaerobe, 8, 287-292.

[36]   Kneifel, W., Rajal, A. and Kulbe, K.D. (2000) In Vitro Growth Behaviour of Probiotic Bacteria in Culture Media with Carbohydrates of Prebiotic Importance. Microbiol Ecology in Health and Disease, 12, 27-34.

[37]   Mathara, J.M., Schillinger, U., Guigas, C., Franz, C., Kutima, P.M., Mbugua, S.K., Shin, H.K. and Holzapfel, W.H. (2008) Functional Characteristics of Lactobacillus spp. from Traditional Maasai Fermented Milk Products in Kenya. International Journal of Food Microbiology, 126, 57-64.

[38]   Nguyen, T.D.T., Kang, J.H. and Lee, M.S. (2007) Characterization of Lactobacillus plantarum PH04, a Potential Probiotic Bacterium with Cholesterol-Lowering Effects. International Journal of Food Microbiology, 113, 358-361.

[39]   Guo, L.-D., Yang, L.-J. and Huo, G.-C. (2011) Cholesterol Removal by Lactobacillus plantarum Isolated from Homemade Fermented Cream in Inner Mongolia of China Czech. Journal of Food Science, 3, 219-225.

[40]   Klaver, F.A.M. and Van Der Meer, R. (1993) The Assumed Assimilation of Cholesterol by Lactobacilli and Bifidobacterium bifidum Is Due to Their Bile Salt-Deconjugating Activity. Applied Environmental Microbiology, 59, 1120-1124.

[41]   Grill, J.P., Cayuela, C., Antoine, J.M. and Schneider, F. (2000) Effects of Lactobacillus amylovorus and Bifidobacterium breve on Cholesterol. Letters in Applied Microbiology, 31, 154-156.

[42]   Begley, M., Hill, C., Cormac, G. and Gahan, M. (2006) Bile Salt Hydrolase Activity in Probiotics. Applied and Environmental Microbiology, 72, 1729-1738.

[43]   Cheeke, P.R. (2000) Actual and Potential Applications of Yucca schidigera and Quillaja saponaria Saponins in Human and Animal Nutrition. Journal of Animal Science, 77, 1-10.

[44]   Ramasamy, K., Abdullah, N., Wong, M.C.V.L., Karuthan, C. and Ho, Y.W. (2010) Bile Salt Deconjugation and Cholesterol Removal from Media by Lactobacillus Strains Used as Probiotics in Chickens. Journal of the Science of Food and Agriculture, 90, 65-69.

[45]   Ridlon, J.M., Kang, D.J. and Hylemon, P.B. (2010) Isolation and Characterization of a Bile Acid Inducible 7α-Dehydroxylating Operon in Clostridium hylemonae TN271. Anaerobe, 16, 137-146.

[46]   Begley, M., Cormac, G.M., Gahan and Hill, C. (2005) The Interaction between Bacteria and Bile. FEMS Microbiology Reviews, 29, 625-651.

[47]   Moser, S.A. and Savage, D.C. (2001) Bile Salt Hydrolase Activity and Resistance to Toxicity of Conjugated Bile Salts Are Unrelated Properties in Lactobacilli. Applied Environmental Microbiology, 67, 3476-3480.

[48]   Brashears, M.M., Gilliland, S.E. and Buck, L.M. (1998) Bile Salt Deconjugation and Cholesterol Removal from Media by Lactobacillus casei. Journal of Dairy Science, 81, 2103-2110.

[49]   Meira, S.M.M., Helfer, V.E., Velho, R.V., Lopes, F.C. and Brandelli, A. (2012) Probiotic Potential of Lactobacillus spp. Isolated from Brazilian Regional Ovine Cheese. Journal of Dairy Research, 79, 119-127.

[50]   Lobo, V., Patil, A., Phatak, A. and Chandra, N. (2010) Free Radicals, Antioxidants and Functional Foods: Impact on Human Health. Pharmacognosy Reviews, 4, 118-126.