شناسایی فنوتیپی و مولکولی باکتری های اسید لاکتیک غیربیماری‌زای جدا شده از ماست گوسفندی محلی

نویسندگان
مبین زمان 1
نفیسه دعوتی 2
مصطفی کرمی 3
1 دانش آموخته کارشناسی ارشد، گروه علوم و صنایع غذایی، دانشکده صنایع غذایی، دانشگاه بوعلی سینا، همدان، ایران
2 دانشیار، گروه علوم و صنایع غذایی، دانشکده صنایع غذایی، دانشگاه بوعلی سینا، همدان، ایران،
3 دانشیار، گروه علوم و صنایع غذایی، دانشکده صنایع غذایی، دانشگاه بوعلی سینا، همدان، ایران، کد پستی
10.22034/FSCT.22.160.1
چکیده
با افزایش تمایل مردم به مصرف محصولات لبنی ارگانیک، همواره خطر سویه‌های میکروبی بیماری‌زا در محصولات محلی وجود دارد. ماست گوسفندی محلی به عنوان یک محصول ارگانیک از باارزش‌ترین محصولات لبنی در ایران است. هدف از این مطالعه تشخیص فنوتیپی و مولکولی باکتری‌های اسید لاکتیک جدا شده از ماست گوسفندی همدان و بررسی خواص بیماری‌زای آن‌ها است. جدایه‌های لاکتیکی از نمونه‌های ماست از نظر فنوتیپی و مولکولی با تکثیر ناحیه 16S rDNA توسط واکنشPCR و به دنبال آن توالی‌یابی شناسایی شدند. خصوصیات بیماریزایی و ایمنی جدایه‌ها شامل مقاومت به آنتی‌بیوتیک، همولیز سلول‌های خونی، فعالیت‌‌‌های آمینواسید دکربوکسیلازی، DNase، و ژلاتینازی بررسی شدند. 47 باکتری از نمونه‌های ماست جدا شدند و تنها 22 جدایه گرم مثبت و کاتالاز منفی بودند. براساس نتایج، جدایه‌ها از نظر مولکولی به عنوان 4 لاکتوباسیلوس هلوتیکوس، 5 انتروکوکوس موری، 2 استرپتوکوکوس سالیواریوس زیرگونه ترموفیلوس، 5 لاکتوباسیلوس هلوتیکوس و 6 لاکتوباسیلوس دلبروکی زیرگونه بولگاریکوس تشخیص داده شدند. شناسایی فنوتیپی همچنین حضور جدایه‌هایی که متعلق به تولید ماست بودند را تایید کرد. ارزیابی خواص بیماری‌زایی گونه‌های مرتبط با تولید ماست، استرپتوکوکوس سالیواریوس زیرگونه ترموفیلوس و لاکتوباسیلوس دلبروکی زیرگونه بولگاریکوس، تایید کرد که تنها دو جدایه از لاکتوباسیلوس دلبروکی زیرگونه بولگاریکوس(HL1, HL2) به عنوان کشت آغازگر برای تولید ماست ایمن هستند
کلیدواژه‌ها
باکتری‌های اسید لاکتیک
آمین‌های بیوژنیک
مقاومت به آنتی بیوتیک

موضوعات

عنوان مقاله English

Phenotypic and molecular identification of the non-pathogenic lactic acid bacteria isolated from the local sheep yogurt

نویسندگان English

Mobin Zaman 1
nafiseh davati 2
mostafa karami 3
1 M.Sc. Graduate, Department of Food Science and Technology, Faculty of Food Industry, Bu-Ali Sina University, Hamedan
2 Associate professor, Department of Food Science and Technology, Faculty of Food Industry, Bu-Ali Sina University, Hamedan, 65178-38695, Iran.
3 Associate professor, Department of Food Science and Technology, Faculty of Food Industry, Bu-Ali Sina University, Hamedan, 65178-38695, Iran
چکیده English

Background: With the growing tendency of people to consume organic dairy products, there is always a risk of pathogenic microbial strains in unhygienic locally prepared products. Local sheep yogurt as an organic product is one of the most valuable dairy products in Iran.

Objectives: The aims of this study were molecular and phenotypic identification of lactic acid bacteria (LAB) isolated from sheep yogurt from Hamadan and investigation of their pathogenic characteristics.

Materials and Methods: The LAB isolated from yogurt samples were identified phenotypically and molecularly by PCR amplification of the 16S rDNA region and subsequent sequencing. The pathogenic and safety characteristics of the isolates including antibiotic resistance, blood hemolysis, amino acid decarboxylase, DNase, and gelatinase activities were then investigated.

Results: Forty-seven bacteria were isolated from yogurt samples, and only 22 gram-positive isolates reacted negatively to catalase. Based on the results, the isolates were molecularly characterized as 4 Lactobacillus helveticus, 5 Enterococcus mori, 2 Streptococcus salivarius subsp. thermophilus, 5 Lactobacillus kalixensis, and 6 Lactobacillus delbrueckii subsp. bulgaricus. Phenotypic identification also confirmed the isolates assigned to S. thermophilus and L. bulgaricus by molecular identification.

Conclusion: The evaluation of the pathogenic characteristics of the species associated with yogurt production, S. thermophilus and L. bulgaricus, confirmed that two isolates (HL1, HL2) of L. bulgaricus are safe as starter culture for yogurt production.




کلیدواژه‌ها English

lactic acid bacteria
Biogenic amines
Antibiotic resistance
1. Omar NB, Castro A, Lucas R, Abriouel H, Yousif NM, Franz CM, et al. Functional and safety aspects of enterococci isolated from different Spanish foods. Systematic and Applied Microbiology. 2004;27(1):118-30.
2. Varela-Ramirez A, Abendroth J, Mejia AA, Phan IQ, Lorimer DD, Edwards TE, et al. Structure of acid deoxyribonuclease. Nucleic acids research. 2017;45(10):6217-27.
3. Barbieri F, Montanari C, Gardini F, Tabanelli G. Biogenic amine production by lactic acid bacteria: A review. Foods. 2019;8(1):17.
4. Ekpenyong M, Asitok A, Odey A, Antai S. Production and activity kinetics of gelatinase by Serratia sp. SLO3. Nigerian Journal of Biopesticides. 2016;1(1):70-82.
5. Piruozpour N, Hanifiyan S. Probiotic potential of Enterococcus species isolated from raw milk and traditional dairy products of Tabriz area. Journal of Food Research. 2019;29(3):13-26.
6. Katiku MM, Matofari JW, Nduko JM. Preliminary evaluation of probiotic properties and safety profile of Lactiplantibacillus plantarum isolated from spontaneously fermented milk, Amabere amaruranu. Heliyon. 2022;8(8).
7. (ISIRI) IoSaIRoI. Milk and milk products –
Guidance on sampling. 3th. revision ed. Iran: Institute of Standards and Industrial Research of Iran (ISIRI); 2008.
8. Benson HJ. Microbiological applications: a laboratory manual in general microbiology. [McGraw-Hill]; 2002.
9. Abdi R, Sheikh-Zeinoddin M, Soleimanian-Zad S. Identification of lactic acid bacteria isolated from traditional Iranian Lighvan cheese. Pakistan Journal of Biological Sciences. 2006;9(1):99-103.
10. Harrigan WF. Laboratory methods in food microbiology. Gulf professional publishing; 1998.
11. Axelsson L. Lactic acid bacteria: classification and physiology. Food Science and Technology-New York-Marcel Dekker-. 2004;139:1-66.
12. Sharma R, Bhaskar B, Sanodiya BS, Thakur GS, Jaiswal P, Yadav N, et al. Probiotic efficacy and potential of Streptococcus thermophiles modulating human health: A synoptic review. J Pharmaceutic Biol Sci. 2014;9:52-8.
13. Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey FA, et al. Bergey's manual of systematic bacteriology: Volume 3: The Firmicutes. Springer Science & Business Media; 2011.
14. Ruiz-Barba JL, Maldonado-Barragán A, Jiménez Díaz R. Small-scale total DNA extraction from bacteria and yeast for PCR applications. 2005.
15. Davati N, Yazdi FT, Zibaee S, Shahidi F, Edalatian MR. Study of lactic acid bacteria community from raw milk of Iranian one humped camel and evaluation of their probiotic properties. Jundishapur journal of microbiology. 2015;8(5).
16. Mousavi S, Kafili T. Antibiotic Resistance of Lactic Acid Bacteria Isolated from Traditional Raw Milk Taleshi Cheese. Journal of Innovation in Food Science and Technology. 2020;11(4):103-14.
17. Davati N. Evaluation of antimicrobial and antibiotic resistance properties of microbial community in a traditional cheese. 2022.
18. Davati N, Hesami S. 16S rRNA metagenomic analysis reveals significant changes of microbial compositions during fermentation from Ewe milk to doogh with antimicrobial activit y. Food Biotechnology. 2021;35(3):179-98.
19. Sandholm E, Sarimo SS. Autolysis of Streptococcus thermophilus. FEMS Microbiology Letters. 1981;11(2-3):125-9.
20. Thomas TD, Crow VL. Lactose and sucrose utilization by Streptococcus thermophilus. 1983.
21. Davati N, Hesami S. Comparison of microbial diversity of ewe's drinking yogurt from somar region nomads using Next-Generation Sequencing and culture dependent molecular methods. Iranian Food Science and Technology Research Journal. 2018;14(5):685-98.
22. Davati N. Isolation and Identification of Indigenous Lactic Acid Bacteria from Traditional Yogurt Produced from Ewe's Milk from Alvand Nomads Region and Evaluation of Their Acidifying Potential. Journal of Food Science and Technology. 2018;15:213-22.
23. Hsueh P-R, Badal RE, Hawser SP, Hoban DJ, Bouchillon SK, Ni Y, et al. Epidemiology and antimicrobial susceptibility profiles of aerobic and facultative Gram-negative bacilli isolated from patients with intra-abdominal infections in the Asia–Pacific region: 2008 results from SMART (Study for Monitoring Antimicrobial Resistance Trends). International journal of antimicrobial agents. 2010;36(5):408-14.
24. Badis A, Guetarni D, Boudjema BM, Henni D, Kihal M. Identification and technological properties of lactic acid bacteria isolated from raw goat milk of four Algerian races. Food Microbiology. 2004;21(5):579-88.
25. Bhardwaj A, Puniya M, Sangu K, Kumar S, Dhewa T. Isolation and biochemical characterization of Lactobacillus species isolated from Dahi. Research & Reviews: A Journal of Dairy Science and Technology. 2012;1:18-31.
26. Bonyadi M, Mojarrad Khangah S, Qanbarov KQ, Gojezadeh M, Dalili Oskuee R. Determination the number of Lactic Acid Bacteria and Yeasts in the combination of traditional yoghurts of villages of East-Azerbaijan-province. Medical Laboratory Journal. 2011;5(2):62-5.
27. Jafari M, Shariatifar N, Khaniki GJ, Abdollahi A. Molecular characterization of isolated lactic acid bacteria from different traditional dairy products of tribes in the Fars province, Iran. Journal of microbiology, biotechnology and food sciences. 2021;11(2):e3621-e.
28. Hajimohammadi Farimani R, Habibi Najafi MB, Fazly Bazzaz BS, Edalatian MR, Bahrami AR, Flórez AB, et al. Identification, typing and functional characterization of dominant lactic acid bacteria strains from Iranian traditional yoghurt. European Food Research and Technology. 2016;242:517-26.
29. Singhal N, Singh NS, Mohanty S, Singh P, Virdi JS. Evaluation of probiotic characteristics of lactic acid bacteria isolated from two commercial preparations available in Indian market. Indian journal of microbiology. 2019;59:112-5.
30. Bernardeau M, Vernoux JP, Henri-Dubernet S, Guéguen M. Safety assessment of dairy microorganisms: the Lactobacillus genus. International journal of food microbiology. 2008;126(3):278-85.
31. Perin LM, Miranda RO, Todorov SD, de Melo Franco BDG, Nero LA. Virulence, antibiotic resistance and biogenic amines of bacteriocinogenic lactococci and enterococci isolated from goat milk. International journal of food microbiology. 2014;185:121-6.
32. Domingos-Lopes M, Stanton C, Ross P, Dapkevicius M, Silva C. Genetic diversity, safety and technological characterization of lactic acid bacteria isolated from artisanal Pico cheese. Food microbiology. 2017;63:178-90.
33. Omafuvbe BO, Enyioha LC. Phenotypic identification and technological properties of lactic acid bacteria isolated from selected commercial Nigerian bottled yoghurt. African Journal of Food Science. 2011;5(6):340-8.