Survival of Lactobacillus Acidophilus as Probiotic Bacteria using Chitosan Nanoparticles


1 Department of Pharmaceutics, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran

2 Pharmaceutical Sciences Research Center, Mazandaran University of Medical Sciences, Sari, Iran

3 Department of Chemical Engineering, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran

4 Department of Food Science and Technology, Sabzevar Branch, Islamic Azad University, Sabzevar, Iran


Chitosan was used for nanoencapsulation of Lactobacillus acidophilus as probiotic bacteria. In vitro experiments were done with the objective of investigating the survival of the bacteria cells in gastro-intestinal conditions. The results demonstrated that the size of chitosan nanoparticles noticeably increases by increasing chitosan concentration from 0.05 to 0.5 g/mL. Encapsulation of the cells caused a decrease at the leakage of probiotic bacteria when compared with free bacteria. However, the number of probiotic cells reduced from 3.27 to 3.23 log CFU/ml in gestural acid condition in contrast to free cells which they approximately dropped from 3.3 to 3 log CFU/ml after 120 min. Good probiotic viability and stability was also obtained by nanocapsulation of Lactobacillus acidophilus in intestinal juice. At this condition, the initial capsulated cells numbers were 3.28 log CFU/ml which after placing in biliary salt condition for 120 min, it reached to 3.23 log CFU/ml. On the other hand, the free bacteria cells reduced from 3.3 to 2.97 log CFU/ml in intestinal environment. Overall, nanocapsulation of probiotic bacteria plays a pivotal role in enhancing the viability and survival of them against gastro- intestinal environmental conditions.


1.     Galdeano, C.M. and Perdigon, G., "Role of viability of probiotic strains in their persistence in the gut and in mucosal immune stimulation", Journal of Applied Microbiology,  Vol. 97, No. 4, (2004), 673-681.

2.     Fitton, N. and Thomas, J.S., "Gastrointestinal dysfunction", Surgery (Oxford),  Vol. 27, No. 11, (2009), 492-495.

3.     Sanders, M.E., "Considerations for use of probiotic bacteria to modulate human health", The Journal of Nutrition,  Vol. 130, No. 2, (2000), 384S-390S.

4.     Vivek, K., "Use of encapsulated probiotics in dairy based foods", International Journal of Food, Agriculture and Veterinary Sciences,  Vol. 3, No. 1, (2013), 188-199.

5.     Mortazavian, A., Razavi, S.H., Ehsani, M.R. and Sohrabvandi, S., "Principles and methods of microencapsulation of probiotic microorganisms", Iranian Journal of Biotechnology,  Vol. 5, No. 1, (2007), 1-18.

6.     Shah, N.P., "Functional cultures and health benefits", International Dairy Journal,  Vol. 17, No. 11, (2007), 1262-1277.

7.     Saranya, S. and Hemashenpagam, N., "Antagonistic activity and antibiotic sensitivity of lactic acid bacteria from fermented dairy products", Advances in Applied Science Research,  Vol. 2, No. 4, (2011), 528-534.

8.     Conway, P.L., "Selection criteria for probiotic microorganisms", Asia Pacific Journal of Clinical Nutrition,  Vol. 5, No., (1996), 10-14.

9.     Ross, R., Fitzgerald, G., Collins, K. and Stanton, C., "Cheese delivering biocultures--probiotic cheese", Australian Journal of Dairy Technology,  Vol. 57, No. 2, (2002), 71.

10.   Reid, G., Sanders, M., Gaskins, H.R., Gibson, G.R., Mercenier, A., Rastall, R., Roberfroid, M., Rowland, I., Cherbut, C. and Klaenhammer, T.R., "New scientific paradigms for probiotics and prebiotics", Journal of Clinical Gastroenterology,  Vol. 37, No. 2, (2003), 105-118.

11.   Lourens-Hattingh, A. and Viljoen, B.C., "Yogurt as probiotic carrier food", International Dairy Journal,  Vol. 11, No. 1, (2001), 1-17.

12.   Favaro-Trindade, C., Heinemann, R. and Pedroso, D., "Developments in probiotic encapsulation", CAB Rev,  Vol. 6, (2011), 1-8.

13.   Chen, M.J. and Chen, K.N., "Applications of probiotic encapsulation in dairy products", Encapsulation and Controlled Release Technologies in Food Systems,  (2007), 83-112.

14.   Gbassi, G.K. and Vandamme, T., "Probiotic encapsulation technology: From microencapsulation to release into the gut", Pharmaceutics,  Vol. 4, No. 1, (2012), 149-163.

15.   Zanjani, M.A.K., Tarzi, B.G., Sharifan, A. and Mohammadi, N., "Microencapsulation of probiotics by calcium alginate-gelatinized starch with chitosan coating and evaluation of survival in simulated human gastro-intestinal condition", Iranian Journal of Pharmaceutical Research: IJPR,  Vol. 13, No. 3, (2014), 843.

16.   Sultana, K., Godward, G., Reynolds, N., Arumugaswamy, R., Peiris, P. and Kailasapathy, K., "Encapsulation of probiotic bacteria with alginate–starch and evaluation of survival in simulated gastrointestinal conditions and in yoghurt", International Journal of Food Microbiology,  Vol. 62, No. 1, (2000), 47-55.

17.   Boh, B., "Developements et applications industrielles des microcapsules", Microencapsulation: Des Sciences Aux Technologies. Paris: Lavoisier,  (2007), 9-22.

18.   Singh, R. and Lillard, J.W., "Nanoparticle-based targeted drug delivery", Experimental and Molecular Pathology,  Vol. 86, No. 3, (2009), 215-223.

19.   Desai, M.P., Labhasetwar, V., Amidon, G.L. and Levy, R.J., "Gastrointestinal uptake of biodegradable microparticles: Effect of particle size", Pharmaceutical Research,  Vol. 13, No. 12, (1996), 1838-1845.

20.   Yeung, T.W., Üçok, E.F., Tiani, K.A., McClements, D.J. and Sela, D.A., "Microencapsulation in alginate and chitosan microgels to enhance viability of bifidobacterium longum for oral delivery", Frontiers in Microbiology,  Vol. 7, (2016).

21.   Anal, A.K. and Singh, H., "Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery", Trends in Food Science & Technology,  Vol. 18, No. 5, (2007), 240-251.

22.   Kouchak, M. and Azarpanah, A., "Preparation and in vitro evaluation of chitosan nanoparticles containing diclofenac using the ion-gelation method", Jundishapur Journal of Natural Pharmaceutical Products,  Vol. 10, No. 2, (2015).

23.   Mohammadpour Dounighi, N., Eskandari, R., Avadi, M., Zolfagharian, H., Mir Mohammad Sadeghi, A. and Rezayat, M., "Preparation and in vitro characterization of chitosan nanoparticles containing mesobuthus eupeus scorpion venom as an antigen delivery system", Journal of Venomous Animals and Toxins Including Tropical Diseases,  Vol. 18, No. 1, (2012), 44-52.

24.   Gilania, S., Najafpour, G., Moghadamniab, A. and Kamaruddinc, A., "Kinetics and isotherm studies of the immobilized lipase on chitosan support".

25.   Kim, J., Muhammad, N., Jhun, B.H. and Yoo, J.-W., "Probiotic delivery systems: A brief overview", Journal of Pharmaceutical Investigation,  Vol. 46, No. 4, (2016), 377-386.

26.   Mohammadpourdounighi, N., Behfar, A., Ezabadi, A., Zolfagharian, H. and Heydari, M., "Preparation of chitosan nanoparticles containing naja naja oxiana snake venom", Nanomedicine: Nanotechnology, Biology and Medicine,  Vol. 6, No. 1, (2010), 137-143.

27.   Werle, M., Takeuchi, H. and Bernkopā€Schnürch, A., "Modified chitosans for oral drug delivery", Journal of Pharmaceutical Sciences,  Vol. 98, No. 5, (2009), 1643-1656.

28.   Pinto, M.G.V., Franz, C.M., Schillinger, U. and Holzapfel, W.H., "Lactobacillus spp. With in vitro probiotic properties from human faeces and traditional fermented products", International Journal of Food Microbiology,  Vol. 109, No. 3, (2006), 205-214.

29.   Fernandez, M., Boris, S. and Barbes, C., "Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract", Journal of Applied Microbiology,  Vol. 94, No. 3, (2003), 449-455.

30.   Chandramouli, V., Kailasapathy, K., Peiris, P. and Jones, M., "An improved method of microencapsulation and its evaluation to protect lactobacillus spp. In simulated gastric conditions", Journal of Microbiological Methods,  Vol. 56, No. 1, (2004), 27-35.

31.   Kafshgari, M.H., Khorram, M., Khodadoost, M. and Khavari, S., "Reinforcement of chitosan nanoparticles obtained by an ionic cross-linking process", Iran Polym J,  Vol. 20, No. 5, (2011), 445-456.

32.   Zhang, H.-l., Wu, S.-h., Tao, Y., Zang, L.-q. and Su, Z.-q., "Preparation and characterization of water-soluble chitosan nanoparticles as protein delivery system", Journal of Nanomaterials,  Vol. 2010, (2010), 1.

33.   Wu, Y., Yang, W., Wang, C., Hu, J. and Fu, S., "Chitosan nanoparticles as a novel delivery system for ammonium glycyrrhizinate", International Journal of Pharmaceutics,  Vol. 295, No. 1, (2005), 235-245.

34.   Zhao, L.-M., Shi, L.-E., Zhang, Z.-L., Chen, J.-M., Shi, D.-D., Yang, J. and Tang, Z.-X., "Preparation and application of chitosan nanoparticles and nanofibers", Brazilian Journal of Chemical Engineering,  Vol. 28, No. 3, (2011), 353-362.