Isolation Identification and Molecular Characterization of Lactic Acid Bacteria From Raw Milk Samples Collected from Erzurum Region

Abstract

Due to its high-water content, milk is an important source of different microbial contents, especially lactic acid bacteria. The aim of this study is to isolate and identify lactic acid bacteria from raw milk samples collected from Erzurum and its surroundings, and to introduce possible new species, or genera, to the taxonomy. For this purpose, DNAs of pure bacterial cultures obtained from 50 raw milk samples collected from producers in Erzurum and its districts were isolated, isolates that differed from each other were selected by rep-PCR, and 11 different species and subspecies [Corynebacterium casei, Enterococcus italicus, E. durans, Lactococcus lactis, Lactococcos lactis subsp. lactis, Lactococcos lactis subsp. hordniae, Lactobacillus paracasei, Leuconostoc lactis, Staphylococcus succinis, Streptococcus parauberis ve S. uberis] in raw milk samples by 16S rRNA sequence analysis. It was concluded that the (GTG)5-PCR method was more successful than BOX-PCR in the identification of lactic acid bacteria isolated from raw milk samples at a species and subspecies level, and the isolate coded TS10, which was 98% similar Streptococcus uberis, may be a new species and should be re-examined with advanced diagnostic techniques.

Keywords

• Raw milk
• Lactic acid bacteria
• Identification
• rep-PCR
• 16S rRNA sequence analysis

Kaynakça

1. Dogan S. Isolation Identification and Molecular Characterization of Lactic Acid Bacteria From Raw Milk Samples Collected from Erzurum Region. Master of Science thesis, Ataturk University, Graduate School of Health Sciences, Erzurum, Turkey; 2021.
2. Gürsoy A. [Internet]; 2011. Süt Kimyası ve Biyokimyası. https://acikders.ankara.edu.tr/course/view.php?id=5685.
3. TÜİK Verileri; 2020. https://data.tuik.gov.tr/Bulten/Index?p=Dis-Ticaret-Istatistikleri-Temmuz-2020-3385. 5 Temmuz 2020.
4. Yalçın H, Özdemir S, Gökalp HY, Kurt A. Ziraat Fakültesi Süt Fabrikasına Farklı Kaynaklardan Gelen İnek Sütlerinde Total, Psikrofilik, Laktik Asit, Koliform Grubu ve S. aureus Bakteri Sayılarının Belirlenmesi. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 1991; 22: 38-45.
5. Yerlikaya O. Starter cultures used in probiotic dairy product preparation and popular probiotic dairy drinks. Food Science Technology, 2014; 34: 221-29.
6. Akbulut S, Baltaci MO, Adiguzel G, Adiguzel A. Identification and potential Biotechnological Characterization of Lactic Acid Bacteria Isolated from White Cheese Samples. Journal of Pure and Applied Microbiology, 2022;16(4):2912-22.
7. Akbulut S. Investigation of the potential use of locally isolated lactic Acid bacteria as probiotics. PhD thesis, Graduate School at Natural and Applied Sciences, Atatürk University, Graduate School at Natural and Applied Sciences, Erzurum, Turkey; 2023.
8. Arslan, S. Isolation, identification and molecular chracterization characterization of lactic acid bacteria from different regions of Turkey white cheese samples. MS thesis, Graduate School at Natural and Applied Sciences, Erzurum, Turkey; 2017.

9. Adiguzel, G. Phenotypic and genotypic characterization of lactic acid bacteria isolated from Turkish dry fermented sausage. PhD. thesis, Graduate School of Health Sciences, Ataturk University, Graduate School of Health Sciences, Erzurum, Turkey; 2008.
10. Sari B, Faiz O, Genc B, Sisecioglu M, Adiguzel A and Adiguzel G. New xylanolytic enzyme from Geobacillus galactosidasius BS61 from a geothermal resource in Turkey. International Journal of Biological Macromolecules, 2018; 119: 1017-26.
11. Gevers D, Huys G and Swings J. Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiology Letters, 2001; 205: 31-36.
12. Saez GD, Hebert EM, Saavedra L, Zarate G. Molecular identification and technological characterization of lactic acid bacteria isolated from fermented kidney beans flours (Phaseolus vulgaris L. and P. coccineus) in northwestern Argentina. Food Research International. 2017;102: 605-15.
13. Erkaya E, Genc B, Akbulut S, Adiguzel G, Omeroglu MA, Ozkan H, Adiguzel A. Bacteriocin Producing Bacteria Isolated from Turkish Traditional Sausage Samples. Journal of Pure and Applied Microbiology, 2020;14(2):1567-76.
14. Hongoh Y, Yuzawa H, Ohkuma M, Kudo T. Evaluation of primers and PCR conditions for the analysis of 16S rRNA genes from a natural environment. FEMS Microbiology Letters, 2003; 221(2): 299-304.
15. Genc B, Tunc MT, Adiguzel A. Characterization of Water-Soluble Extracellular Polysaccharide from Aeribacillus pallidus IM17, Indian Journal of Microbiology, https://doi.org/10.1007/s12088-023-01178-2
16. Nadaroglu H, Mosber G, Alayli Gungor A, Adıguzel G, Adiguzel A. Biodegradation of some azo dyes from wastewater with laccase from Weissella viridescens LB37 immobilized on magnetic chitosan nanoparticles, Journal of Water Process Engineering, 2019; 31; 100866.
17. Altinok F, Albayrak S, Arslan NP, Taskin M, Aygun E, Sisecioglu M, Adiguzel A. Application of Anoxybacillus gonensins UF7 lipase as a catalyst for biodiesel production from waste frying oils. Fuel, 2023; 334(1): 126672.
18. Heredia Y, Castro J. Antimicrobial activity and partial characterization of bacteriocin-like inhibitory substances produced by Lactobacillus spp. isolated from artisanal Mexican cheese. Journal of Dairy Science, 2015;12: 8285-93.
19. Ledina T, Mohar P, Golop M. Tetracycline resistance in Lactobacilli isolated from Serbian traditional raw milk cheeses. Journal of Food Scıence and Technology-Mysore, 2018;55: 1426-34.
20. Baltaci MO, Omeroglu MA, Albayrak S, Adiguzel G, Adiguzel A. Production of Endoglucanase by Exiguobacterium mexicanum OB24 Using Waste Melon Peels as Substrate. Anais da Academia Brasileira de Ciências, 2022;94:e20220151.
21. Lusk TS, Ottesen AR, White JR, Allard MW, Brown EW, Kase JA. Characterization of microflora in Latin-style cheeses by next generation sequencing technology. BMC Microbiology, 2012;12: 254.
22. Quilodrán-Vega, Villena SR, Valdebenito J, Salas J, Parra MJ, Ruiz C, Kitazawa A, García HA. Isolation of lactic acid bacteria from swine milk and characterization of potential probiotic strains with antagonistic effects against swine-associated gastrointestinal pathogens. Canadian Journal of Microbiology, 2016;6: 514- 24.
23. Gebru YA, Sbhatu DB. Isolation and characterization of probiotic LAB from kimchi and spontaneously fermented teff (Eragrostis tef (Zucc.) Trotter) batter: their effects on phenolic content of teff during fermentation. Hindawi BioMed Research International, 2020;2020: 4014969.
24. Kirmaci HA. Isolation,_molecular characterization of lactic acid bacteria in traditional Urfa cheese and possible use as a starter culture. PhD. thesis, Graduate School at Natural and Applied Sciences, Harran University, Sanliurfa, Turkey, 2010.
25. Fortina MG, Ricci G, Mora D, Manachini PL. Molecular analysis of artisanal Italian cheeses reveals Enterococcus italicus sp. nov. International Journal of Systematic and Evolutionary Microbiology, 2004;54: 1717-21.
26. Odierno L, Calvinho L, Traverssa P. Conventional identification of Streptococcus uberis isolated from bovine mastitis in Argentinean dairy herds. Journal of Dairy Science, 2006; 89: 3886-90.
27. Tanguler H. Identification of predominant lactic acid bacteria isolated from salgam beverage and improvement of its production technique. PhD, thesis, Graduate School at Natural and Applied Sciences, Cukurova University, Adana, Turkey, 2010.
28. Kuikui N, Yanping W, Yimin C. Natural Lactic Acid Bacteria Population and Silage Fermentation of Whole-crop Wheat. Asian Australasian Journal of Animal Sciences. 2015; 28: 1123-32.
29. Brennan NM, Brown R, Goodfellow M, Cogan TM. Corynebacterium mooreparkense sp. nov. and Corynebacterium casei sp. nov., isolated from the surface of a smear-ripened cheese. International Journal of Systematic and Evolutionary Microbiology, 2001; 51: 843-52.
30. Sánchez C, Neves AR, Cavalheiro J, Santos H. Contribution of citrate metabolism to the growth of Lactococcus lactis CRL264 at low pH. Applied and Environmental Microbiology, 2008; 74: 1136-44.