Isolation and Identification of Lactobacillus Acidophilus Probiotic Bacteria from Traditional Tarkhineh Food and Their Ability to Lower Cholesterol and Triglyceride Levels in vitro
Subject Areas : MicrobiologyP. Afshar 1 , K. Amini 2 , H. Mohajerani 3 , S. Saki 4
1 - PhD Student of the Department of Microbiology, Faculty of Sciences, Arak Branch, Islamic Azad University, Arak, Iran.
2 - Assistant Professor of the Department of Microbiology, Faculty of Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran
3 - Assistant Professor of the Department of Biology, Arak Branch, Islamic Azad University, Arak, Iran.
4 - 4. Assistant Professor of the Department of Medical Laboratory Sciences, Faculty of Medical Sciences, Islamic Azad University, Arak Branch, Arak, Iran.
Keywords: Cholesterol, Lactobacillus acidophilus, Probiotics, Tarkhineh,
Abstract :
Introduction: Isolation and identification of probiotic strains from native foods can lead to finding strains with unique capabilities. The aim of this study was to isolate, identify and evaluate the ability of Lactobacillus acidophilus bacteria isolated from traditional Tarkhineh food in lowering cholesterol and triglyceride levels in vitro.Materials and Methods: Native strains of Lactobacillus acidophilus were isolated from a local food called Tarkhineh using culture medium and identified based on phenotypic characteristics, standard biochemical tests and 16SrRNA sequencing. In the next step, the strains were examined for probiotic properties such as resistance to acid and bile and sensitivity to antibiotics. Cholesterol and triglyceride lowering activity of isolates in culture medium was also investigated by o-phthaldehyde method.Results: The results showed that 16 strains of Lactobacillus acidophilus were isolated from Tarkhineh samples, among which four strains (S3, S12, S6, S5) showed 30 to 50% resistance, three strains (S7, S4, S10). had 50 to 75% resistance and 2 strains (S11, S1) had 75 to 100% growth in acidic MRS. In addition, strains S7 and S1 showed a growth rate of 0.3 to 0.5% and strains S4, S3, S11 grew at 0.7% bile salt in MRS medium. The highest cholesterol uptake was observed in S11 and S1 strains at 64± 0.23 and 60±0.18 (p<0.01) respectively. Strains S1 and S11 applied 72% ± 0.64 and 61 ± 0.25% triglyceride reduction respectively.Conclusion: The data of this study showed that the native strains of Lactobacillus acidophilus isolated from the local food of Tarkhineh had the potential to absorb and reduce cholesterol and triglycerides of the culture medium. These strains can be studied in laboratory animals as probiotics capable of absorbing and removing fat.
Aguilar-Toalá, J., Garcia-Varela, R., Garcia, H., Mata-Haro, V., González-Córdova, A., Vallejo-Cordoba, B. & Hernández-Mendoza, A. (2018). Postbiotics: An evolving term within the functional foods field. Trends in Food Science & Technology, 75, 105-114.
Benjamin, E. J., Muntner, P., Alonso, A., Bittencourt, M. S., Callaway, C. W., Carson, A. P., Chamberlain, A. M., Chang, A. R., Cheng, S. & Das, S. R. (2019). Heart disease and stroke statistics—2019 update: a report from the American Heart Association. Circulation, 139(10), e56-e528.
Bhat, B. & Bajaj, B. K. (2020). Multifarious cholesterol lowering potential of lactic acid bacteria equipped with desired probiotic functional attributes. 3 Biotech, 10(5), 1-16.
Birch, C. S. & Bonwick, G. A. (2019). Ensuring the future of functional foods. International Journal of Food Science & Technology, 54(5), 1467-1485.
Ejtahed, H., Mohtadi-Nia, J., Homayouni-Rad, A., Niafar, M., Asghari-Jafarabadi, M., Mofid, V. & Akbarian-Moghari, A. (2011). Effect of probiotic yogurt containing Lactobacillus acidophilus and Bifidobacterium lactis on lipid profile in individuals with type 2 diabetes mellitus. Journal of dairy science, 94(7), 3288-3294.
El-Saadony, M. T., Alagawany, M., Patra, A. K., Kar, I., Tiwari, R., Dawood, M. A., Dhama, K. & Abdel-Latif, H. M. (2021). The functionality of probiotics in aquaculture: an overview. Fish & Shellfish Immunology, 117, 36-52.
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Izadi, M., Fooladi, M. H., SHarifi Sirchi, G. & Amini, J. (2010). Isolation of Lactobacillus acidophilus from Sharbabk city yoghurt and its molecular characterization. Agricultural Biotechnology Journal, 2(2), 1-12.
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Kim, E., Chang, H. C. & Kim, H.Y. (2020). Complete genome sequence of Lactobacillus plantarum EM, a putative probiotic strain with the cholesterol-lowering effect and antimicrobial activity. Current Microbiology, 77(8) 1871-1882.
Ma, C., Zhang, S., Lu, J., Zhang, C., Pang, X. & Lv, J. (2019). Screening for cholesterol-lowering probiotics from lactic acid bacteria isolated from corn silage based on three hypothesized pathways. International Journal of Molecular Sciences, 20(9), 2073.
Mayrhofer, S., Domig, K. J., Mair, C., Zitz, U., Huys, G. & Kneifel, W. (2008). Comparison of broth microdilution, Etest, and agar disk diffusion methods for antimicrobial susceptibility testing of Lactobacillus acidophilus group members. Applied and Environmental Microbiology, 74(12), 3745-3748.
Majeed, M., Majeed, S., Nagabhushanam, K., Arumugam, S., Beede, K. & Ali, F. (2019). Evaluation of the in vitro cholesterol‐lowering activity of the probiotic strain Bacillus coagulans MTCC 5856. International Journal of Food Science & Technology, 54(1), 212-220.
Nami, Y., Bakhshayesh, R. V., Manafi, M. & Hejazi, M. A. (2019). Hypocholesterolaemic activity of a novel autochthonous potential probiotic Lactobacillus plantarum YS5 isolated from yogurt. LWT, 111, 876-882.
Pourrajab, B., Fatahi, S., Dehnad, A., Varkaneh, H. K. & Shidfar, F. (2020). The impact of probiotic yogurt consumption on lipid profiles in subjects with mild to moderate hypercholesterolemia: a systematic review and meta-analysis of randomized controlled trials. Nutrition, Metabolism and Cardiovascular Diseases, 30(1), 11-22.
Rerksuppaphol, S. & Rerksuppaphol, L. (2015). A randomized double-blind controlled trial of Lactobacillus acidophilus plus Bifidobacterium bifidum versus placebo in patients with hypercholesterolemia. Journal of clinical and diagnostic research: JCDR, 9(3), KC01.
Shemshad, N., Roozbeh Nasiraie, L. & Majidzadeh Heravi, R. (2021). Isolation of Probiotic Lactobacilli Bacteria from Traditional Naein Dairy Product (Koome). Iranian Journalof Medical Microbiology, 15(1), 85-106.
Sivamaruthi, B. S., Kesika, P. & Chaiyasut, C. (2019). A mini-review of human studies on cholesterol-lowering properties of probiotics. Scientia Pharmaceutica, 87(4), 26.
Mohammadi, H. & Tukmechi, A. (2018). A. In vitro evaluation of cholesterol lowering effects of isolated Lactobacillus probiotics from cattle and buffalo feces, Journal of veterinaty microbiology,37(8),31.[In persian]
_||_Aguilar-Toalá, J., Garcia-Varela, R., Garcia, H., Mata-Haro, V., González-Córdova, A., Vallejo-Cordoba, B. & Hernández-Mendoza, A. (2018). Postbiotics: An evolving term within the functional foods field. Trends in Food Science & Technology, 75, 105-114.
Benjamin, E. J., Muntner, P., Alonso, A., Bittencourt, M. S., Callaway, C. W., Carson, A. P., Chamberlain, A. M., Chang, A. R., Cheng, S. & Das, S. R. (2019). Heart disease and stroke statistics—2019 update: a report from the American Heart Association. Circulation, 139(10), e56-e528.
Bhat, B. & Bajaj, B. K. (2020). Multifarious cholesterol lowering potential of lactic acid bacteria equipped with desired probiotic functional attributes. 3 Biotech, 10(5), 1-16.
Birch, C. S. & Bonwick, G. A. (2019). Ensuring the future of functional foods. International Journal of Food Science & Technology, 54(5), 1467-1485.
Ejtahed, H., Mohtadi-Nia, J., Homayouni-Rad, A., Niafar, M., Asghari-Jafarabadi, M., Mofid, V. & Akbarian-Moghari, A. (2011). Effect of probiotic yogurt containing Lactobacillus acidophilus and Bifidobacterium lactis on lipid profile in individuals with type 2 diabetes mellitus. Journal of dairy science, 94(7), 3288-3294.
El-Saadony, M. T., Alagawany, M., Patra, A. K., Kar, I., Tiwari, R., Dawood, M. A., Dhama, K. & Abdel-Latif, H. M. (2021). The functionality of probiotics in aquaculture: an overview. Fish & Shellfish Immunology, 117, 36-52.
Granato, D., Barba, F. J., Bursać Kovačević, D., Lorenzo, J. M., Cruz, A. G. & Putnik, P. (2020). Functional foods: Product development, technological trends, efficacy testing, and safety. Annual Review of Food Science and Technology, 11, 93-118.
Izadi, M., Fooladi, M. H., SHarifi Sirchi, G. & Amini, J. (2010). Isolation of Lactobacillus acidophilus from Sharbabk city yoghurt and its molecular characterization. Agricultural Biotechnology Journal, 2(2), 1-12.
Kerry, R. G., Patra, J. K., Gouda, S., Park, Y., Shin, H.S. & Das, G. (2018). Benefaction of probiotics for human health: A review. Journal offood and drug analysis, 26(3), 927-939.
Kim, E., Chang, H. C. & Kim, H.Y. (2020). Complete genome sequence of Lactobacillus plantarum EM, a putative probiotic strain with the cholesterol-lowering effect and antimicrobial activity. Current Microbiology, 77(8) 1871-1882.
Ma, C., Zhang, S., Lu, J., Zhang, C., Pang, X. & Lv, J. (2019). Screening for cholesterol-lowering probiotics from lactic acid bacteria isolated from corn silage based on three hypothesized pathways. International Journal of Molecular Sciences, 20(9), 2073.
Mayrhofer, S., Domig, K. J., Mair, C., Zitz, U., Huys, G. & Kneifel, W. (2008). Comparison of broth microdilution, Etest, and agar disk diffusion methods for antimicrobial susceptibility testing of Lactobacillus acidophilus group members. Applied and Environmental Microbiology, 74(12), 3745-3748.
Majeed, M., Majeed, S., Nagabhushanam, K., Arumugam, S., Beede, K. & Ali, F. (2019). Evaluation of the in vitro cholesterol‐lowering activity of the probiotic strain Bacillus coagulans MTCC 5856. International Journal of Food Science & Technology, 54(1), 212-220.
Nami, Y., Bakhshayesh, R. V., Manafi, M. & Hejazi, M. A. (2019). Hypocholesterolaemic activity of a novel autochthonous potential probiotic Lactobacillus plantarum YS5 isolated from yogurt. LWT, 111, 876-882.
Pourrajab, B., Fatahi, S., Dehnad, A., Varkaneh, H. K. & Shidfar, F. (2020). The impact of probiotic yogurt consumption on lipid profiles in subjects with mild to moderate hypercholesterolemia: a systematic review and meta-analysis of randomized controlled trials. Nutrition, Metabolism and Cardiovascular Diseases, 30(1), 11-22.
Rerksuppaphol, S. & Rerksuppaphol, L. (2015). A randomized double-blind controlled trial of Lactobacillus acidophilus plus Bifidobacterium bifidum versus placebo in patients with hypercholesterolemia. Journal of clinical and diagnostic research: JCDR, 9(3), KC01.
Shemshad, N., Roozbeh Nasiraie, L. & Majidzadeh Heravi, R. (2021). Isolation of Probiotic Lactobacilli Bacteria from Traditional Naein Dairy Product (Koome). Iranian Journalof Medical Microbiology, 15(1), 85-106.
Sivamaruthi, B. S., Kesika, P. & Chaiyasut, C. (2019). A mini-review of human studies on cholesterol-lowering properties of probiotics. Scientia Pharmaceutica, 87(4), 26.
Mohammadi, H. & Tukmechi, A. (2018). A. In vitro evaluation of cholesterol lowering effects of isolated Lactobacillus probiotics from cattle and buffalo feces, Journal of veterinaty microbiology,37(8),31.[In persian]