Isolation and identification of indigenous Lactobacilli in traditional dairy products in Iran

Ghobadi Dana, M.; Hatef Salmanian, A.; Yakhchali, B.

doi:10.22101/JRIFST.2012.06.21.123

Isolation and identification of indigenous Lactobacilli in traditional dairy products in Iran

Document Type : Original Paper

Authors

¹ Assistant Professor, Department of Microbiology, Standard Research Institute

² Associated Professor, Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology

³ Associated Professor, Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology

10.22101/JRIFST.2012.06.21.123

Abstract

One hundred and twelve traditional dairy samples including yoghurt, doogh and raw milk were collected from Kermanshah, Kurdistan, Lorestan, Ilam and Markazi provinces in Iran. The bacteria were isolated based on standard procedure. The samples were sub-cultured on selective media, typical colonies were examined microscopically and finally ninety three bacteria were selected for further analyses. The isolated bacteria were characterized by fermentation of 19 sugars and growing in 15 and 45 ºC, nitrate reduction, gelatin solidification and esculin degradation. In order to confirm the biochemical results, molecular analysis such as amplification of lactobacillus genus specific fragment with specific primer was carried out. The results showed that all isolates belong to lactobacillus. Four lactobacillus with specific properties were further identified by amplification and sequencing of 16S rDNA gene. These genes were submitted to gene bank in NCBI. The phylogenetic tree was constructed after comparing with other 16S rDNA genes. Results showed that two strains were similar to Lactobacillus crustorum which was recorded recently.

Keywords

References

اجدانی، ه. و ملکزاده، ف.1380. شناسایی سویه های استارتر های بومی مولد ماست ، جداسازی نگهداری و بررسی نقش هریک از آنها: دانشگاه آزاد اسلامی واحد قم.

بارویی، الف. و کرباسی، م.1379. ارزیابی باکتریهای اسیدلاکتیک موجود در فراورده های لبنی سنتی (پنیر) برای تولید استارتر در مقایسه با استارترهای تجاری: دانشگاه شیراز.

جان‌جانی، م.1381. بررسی نقش و اهمیت استارتر در فراورده‌های پنیر چدار آنزیم لیپولیتیک: جهادسازندگی خراسان. 4- طاهری، الف. و احسانی، م. 1384. بررسی امکان تولید ونگهداری نوشیدنی تخمیری پروبیوتیک با استفاده از کشت باکتریایی گرما دوست و میاندوست دانشگاه آزاد اسلامی واحد علوم وتحقیقات تهران.

فاضلی، ع.1382. آزمایشگاهی ‏‎‎‏انتروتوکسیکوژنیک توسط لاکتوباسیل کازئی در مدل موش E-Coli بررسی کاهش کلونیزاسیون: دانشگاه علوم پزشکی اصفهان.

کازرونی، الف. و احسانی، م.1382 .تاثیر افزودن باکتریهای استارتر حرارت دیده و پروتئوزهای تجارتی در تسریع رسانیدن پنیر سفید ایرانی - مؤسسه تحقیقات علوم دامی.

گمرک .1383 .اطلاعات آمار صادرات و واردات کشور.

محمدی، م.1383. بررسی اثر مهاری لاکتوباسیل اسیدفیلوس بر روی سویه های بیماریزای اشرشیاکلای درطیور.

نوحی، الف.1367. میکروبیولوژی عمومی: موسسه انتشارات و چاپ دانشگاه تهران.

Ayhan, K., Durlu-ozkaya, F., & Tunail, N. 2005. Commercially important characteristics of Turkish origin domestic strains of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. International Journal of Dairy Technology, 58: 150-157.

Bergeys. 2005. Manual of determinative bacteriology.

Beukes, E. M., Bester, B. H., & Mostert, J. F. 2001. The microbiology of South African traditional fermented milks. International Journal of Food Microbiology, 63: 189-197.

Carl, B. 1999. Lactobacillus. Academic press.

Chammas, G. I., Saliba, R., Corrieu, G., & Beal, C. 2006. Characterisation of lactic acid bacteria isolated from fermented milk "laban". International Journal of Food Microbiology, 110: 52-61.

FAO/WHO. 2002. Guidelines for the evaluation of probiotics in food London Ontario.

Fitzsimons, N.A., Cogan, T.M., Condon, S. & Beresford, T. 1999. Phenotypic and genotypic characterization of non-starter lactic acid bacteria in mature cheddar cheese. Applied and Environmental Microbiology, 65: 3418 - 3426.

Ghobadi dana, M., Salmanian, A. H., Yakhchali & Rastegar Jazii, F. 2010. High folate production by naturally occurring lactobacillus Spp. with probiotics potential, isolated from dairy products in Ilam and Lorestan provinces of Iran. African Journal of iotechnology, 9: 5383-5391.

Ghobadi Dana, M., Yakhchali, B., Salmanian, A.H., & Rastgarjazi Jazii, F. 2011. High-level acetaldehyde production through a putative acetoin dehydrogenase complex by indigenous lactobacillus strain obtained from traditional dairy products of Iran. African Journal of Microbiology Research, 5(25): 4398-4405.

Ghobadi Dana, M., Salmanian, A.H.,Yakhchali, B. & Rastgarjazi Jazii, F. 2010. High-level acetaldehyde production by wild type lactobacillus strains isolated from traditional dairy products of Iran. 22nd International ICFMH Symposium, FoodMicro 2010. Denmark.

Hammes, V. 1995. The genus Lactobacillus, London, Chapman and Hal.l.

IDF149, I. 2008 Milk products -Acidifying starter cultures- Standard of identify.

Scheirlinck, I., Meulen, R.V., Schoor, A.V., Huys, G., Vandamme, P., Vuyst, L.D. & Vancanneyt, M. 2007. Lactobacillus crustorum sp. nov., isolated from two traditional Belgian wheat sourdoughs. International Journal of Systematic and Evolutionary Microbiology, 57: 1461–1467.

Jokovic, N., Nikolic, M., Begovic, J., Jovcic, B., Savic, D., & Topisirovic, L. 2008. A survey of the lactic acid bacteria isolated from Serbian artisanal dairy product kajmak. International Journal of Food Microbiology, 127: 305-311.

Klaenhammer, T., Altermann, E., Arigoni, F., Bolotin, A., Breidt, F., Broadbent, J., Cano, R., Chaillou, S., Deutscher, J., Gasson, M., Van De Guchte, M., Guzzo, J., Hartke, A., Hawkins, T., Hols, P., Hutkins, R., Kleerebezem, M., Kok, J., Kuipers, O., Lubbers, M., Maguin, E. Mckay, L., Mills, D., Nauta, A., Overbeek, R., Pel, H., Pridmore, D., Saier, M., Van Sinderen, D., Sorokin, A., Steele, J., O'Sullivan, D., DE VOS, W., Weimer, B., Zagorec, M., & Siezen, R. 2002. Discovering lactic acid bacteria by genomics. Antonie van Leeuwenhoek. International Journal of General and Molecular Microbiology, 82: 29-58.

Klein, G., Pack, A., Bonaparte, C., & Reuter, G. 1998. Taxonomy and physiology of probiotic lactic acid bacteria. International Journal of Food Microbiology, 41: 103-125.

Kostinek, M., Specht, I., Edward, V.A., Pinto, C., Egounlety, M., Sossa, C., Mbugua, S., Dortu, C., Thonart, P., Taljaard, L., Mengu, M., Franz, C.M.A.P & Holzapfel, W.H. 2007. Characterization and biochemical properties of predominant lactic acid bacteria from fermenting cassava for selection as starter cultures. International Journal of Food Microbiology, 114(3): 342-351.

Krieg N R., Holt, J. G. 1984. Bergeys Manual of Systematic Bacteriology, Williams and Wikins.

Kwon, H. S., Yang, E. H., Yeon, S. W., Kang, B. H., & Kim, T. Y. 2004. Rapid identification of probiotic Lactobacillus species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA. FEMS Microbiology Letters, 239: 267-275.

Mathara, J. M., Schillinger, U., Kutima, P. M., Mbugua, S. K., & Holzapfel, W. H. 2004. Isolation, identification and characterisation of the dominant microorganisms of kule naoto: The Maasai traditional fermented milk in Kenya. International Journal of Food Microbiology, 94: 269-278.

Nour, M. 1998. 16S-23S and 23S-5S intergenic spacer regions of lactobacilli: Nucleotide sequence, secondary structure and comparative analysis. Research in Microbiology, 149: 433-448.

Pourahmad, R., & Assadi, M. M. 2007. Use of isolated autochthonous starter cultures in yogurt production. International Journal of Dairy Technology, 60: 259-262.

Rossetti, L., Fornasari, M. E., Gatti, M., Lazzi, C., Neviani, E., & Girraffa, G. 2008. Grana Padano cheese whey starters: microbial composition and strain distribution. International Journal of Food Microbiology, 127: 168-171.

Schillinger, U. 2005. Identification of predominant Lactic acid bacteria isolated from traditionally fermented vegetable products of the Eastern Himalayas. International Journal of Food Microbiology, 105: 347-356.

Standard, I. 2003. Yogurt-identification of characteristic microorganisms (Lactobacillus delbrueckii subsp.bulgaricus and Streptococcus thermophilus). ISO9232 IDF146.

Standard, I. 2005. Foodstuffs- Methods of analysis for the detection of genenetically modified organisms and derived product-nucleic acid extraction. IN ISO21. Ed. 571.

Standard, I. 2006. Milk products - enumeration of presumptive Lactobacillus acidophilus on a selective medium - Colony-count technique at 37 °C. IN IDF192, I.

Ward, L. J. H., & Timmins, M. J. 1999. Differentiation of lactobacillus casei, lactobacillus paracasei and lactobacillus rhamnosus by polymerase chain reaction. Letters in Applied Microbiology, 29: 90-92.

Watanabe, K., Fujimoto, J., Sasamoto, M., Dugersuren, J., Tumursuh, T., & Demberel, S. 2008 Diversity of lactic acid bacteria and yeasts in Airag and Tarag, traditional fermented milk products of Mongolia. World Journal of Microbiology and Biotechnology, 24: 1313-1325.