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Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi

Yıl 2021, Cilt: 14 Sayı: 2, 238 - 246, 30.06.2021
https://doi.org/10.30607/kvj.895295

Öz

Boza, insan sağlığı için yararlı mikroorganizmaları içeren fermente bir içecektir. Çalışmamızda boza üretiminde ham madde olarak kullanılan (mısır unu, buğday unu, mayşe) ve boza fermantasyonunun 1. günü, 3. günü ve 4. gün son ürün boza’nın içerdiği mikrobiyota Yeni Nesil DNA Dizileme yöntemi ve metagenomik analiz ile ortaya çıkarılmıştır. Örneklerden doğrudan cins düzeyinde yapılan analiz sonucunda, mısır unu ve buğday ununda dominant olarak Streptophyta ve Pleomorphobacterium bulunurken; bozanın 1. gün, 3. gün ve son ürün ile boza mayasında dominant bakterilerin Leuconostoc ve Lactococcus cinsine ait olduğu tespit edilmiştir. Ön zenginleştirme yapılan örneklerin analizinde, mısır ununda dominant bakteriler Enterococcus, Klebsiella ve Micromonospora, buğday ununda ise Pantoea ve Bacillus olduğu, boza mayası, 1. gün boza, 3. gün boza ve satışa sunulan son üründe dominant bakteri Lactococcus olarak belirlenmiştir. Çalışmamızda örnekler arasındaki bakteriyel çeşitlilik, benzerlik ve farklılıklar Principal Coordinate Analiz ve dendrogram oluşturulması ile ortaya konmuştur. Boza üretiminde kullanılan ham maddelerin bozanın fermantasyon aşamalarındaki ürünler ile fermantasyon sürecinde mikrobiyotasına nasıl değiştiği ve son ürüne olan katkıları, DNA düzeyinde yapılan metagenomik analizler ile belirlenmiştir.

Teşekkür

Çalışma kapsamında boza ve ham madde örneklerini sağlayan Sayın Mustafa Günnaz’a ve laboratuvar alt yapısının kullanıldığı Konya Gıda ve Tarım Üniversitesi, Kit-ARGEM Araştırma Merkezi’ne teşekkür ederiz.

Kaynakça

  • Altay F, Karbancıoglu-Güler F, Daskaya-Dikmen C, Heperkan D. A review on traditional Turkish fermented non-alcoholic beverages: Microbiota, fermentation process and quality characteristics. Int J Food Microbiol. 2013; 167(1): 44-56. https://doi.org/10.1016/j.ijfoodmicro.2013.06.016
  • Arici M, Daglioglu O. Boza: A lactic acid fermented cereal beverage as a traditional Turkish food. Food Rev Int. 2002; 18(1): 39-48. https://doi.org/10.1081/FRI-120003416
  • Baschali A, Tsakalidou E, Kyriacou A, Karavasiloglou N, Matalas AL. Traditional low-alcoholic and non-alcoholic fermented beverages consumed in European countries: A neglected food group. Nutr Res Rev. 2017; 30(1): 1-24. https://doi.org/10.1017/S0954422416000202
  • Bayat G, Yıldız G. The special fermented Turkish drink: Boza. Journal of Tourism and Gastronomy Studies 2019; 7(4): 2438-2446. https://doi.org/10.21325/jotags.2019.480
  • Bliss ES, Whiteside E. The gut-brain axis, the human gut microbiota and their integration in the development of obesity. Fron Physiol. 2018; 9: 900. https://doi.org/10.3389/fphys.2018.00900
  • Borcaklı M, Öztürk T, Yeşilada E. Cereal source amd microbial consortia of the starter culture influence the chemical composition and physicochemical characteristics of boza. Turk J Agric For. 2018; 42: 412-422. https://doi.org/10.3906/tar-1802-3
  • Botes A, Todorov SD, von Mollendorff JW, Botha A, Dicks LM. Identification of LAB and yeast from boza. Process Biochem. 2007; 42(2): 267-270. https://doi.org/10.1016/j.procbio.2006.07.015
  • Cholakov R, Tumbarski Y, Yanakieva V, Dobrev I, Salim Y, Denkova Z. Antimicrobial acitivity of Leuconostoc lactis strain BT17, isolated from a spontaneously fermented cereal beverage (Boza). J Microbiol Biotech Food Sci. 2017; 7(1): 47-49. https://doi.org/10.15414/jmbfs.2017.7.1.47-49
  • Çakır C. Tarihi, Etimolojisi ve Edebiyatı ile Boza Kitabı, Türk Dünyası Araştırmaları Vakfı Yayınevi, 1. Baskı, İstanbul. 2019; pp. 17-19.
  • De Vuyst L, Leroy F. Functional role of yeasts, lactic acid bacteria, and acetic acid bacteria in cocoa fermentation processes. FEMS Microbiol Rev. 2020; 44(4): 432-453. https://doi.org/10.1093/femsre/fuaa014
  • Dogan M, Ozpinar H. Investigation of probiotic features of bacteria isolated from some food products. Kafkas Univ Vet Fak Derg. 2017; 23(4): 555-562. https://doi.org/10.9775/kvfd.2016.17273
  • Doğan M, Tekiner İH. Extracellular phytase activites of lactic acid bacteria in sourdough mix prepared from traditionally produced boza as starter culture. Food and Health. 2020; 6(2): 117-127. https://doi.org/10.3153/FH20013
  • Gibson GR, Hutkins R, Sanders ME, Prescott SL, Reimer RA, Salminen SJ, Scott K, Stanton C, Swanson KS, Cani PD, Verbeke K, Reid G. The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat Rev Gastroenterol Hepatol. 2017; 14(8): 491-502. https://doi.org/10.1038/nrgastro.2017.75
  • Gotcheva V, Pandiella SS, Angelov A, Roshkova ZG, Webb C. Microflora identification of the Bulgarian cereal-based fermented beverage boza. Process Biochem. 2000; 36(1-2): 127-130. https://doi.org/10.1016/S0032-9592(00)00192-8
  • Gotcheva V, Pandiella SS, Angelov A, Roshkova Z, Webb C. Monitoring the fermentation of the traditional Bulgarian beverage boza. Int J Food Sci Tech. 2001; 36: 129-134. https://doi.org/10.1046/j.1365-2621.2001.00429.x
  • Hancioğlu Ö, Karapinar M. Microflora of Boza, a traditional fermented Turkish beverage. Int J Food Microbiol. 1997; 35(3): 271-274. https://doi.org/10.1016/S0168-1605(96)01230-5
  • Kıvanç M, Yılmaz M, Çakır E. Isolation and identification of lactic acid bacteria from boza, and their microbial activity against several reporter strains. Turk J Biol. 2011; 35(3): 313-324. https://doi.org/10.3906/biy-0906-67
  • Levent H, Cavuldak ÖA. Geleneksel Fermente Bir İçecek: Boza. Akademik Gıda. 2017; 15(3): 300-307. https://doi.org/10.24323/akademik-gida.345273
  • Lindgren SW, Dobrogosz WJ. Antagonistic activities of lactic acid bacteria in food and feed fermentations. FEMS Microbiol Rev. 1990; 87: 149-164. https://doi.org/10.1111/j.1574-6968.1990.tb04885.x
  • Liu D, Ainsworth AJ, Austin FW, Lawrence ML. Use of PCR primers derived from a putative transcriptional regulator gene for species-specific determination of Listeria monocytogenes. Int J Food Microbiol. 2004; 91(3): 297-304. https://doi.org/10.1016/j.ijfoodmicro.2003.07.004
  • Marco ML, Sanders ME, Gänzle M, Arrieta MC, Cotter PD, De Vuyst L, Hill C, Holzapfel W, Lebeer S, Merenstein D, Reid G, Wolfe BE, Hutkins R. The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on fermented foods. Nat Rev Gastroenterol Hepatol. 2021. https://doi.org/10.1038/s41575-020-00390-5
  • Osimani A, Garofalo C, Aquilant L, Milanović V, Clementi F. Unpasteurised commercial boza as a source of microbial diversity. Int J Food Microbiol. 2015; 194: 62-70. http://dx.doi.org/10.1016/j.ijfoodmicro.2014.11.011
  • Palamutoğlu Mİ, Baş M. Traditional Fermented Foods of Turkey. J Health Sci Res. 2020; 2(3): 200-220. https://doi.org/10.46413/boneyusbad.790343
  • Rovella V, Rodia G, Di Daniele F, Cardillo C, Campia U, Noce A, Candi E, Della-Morte D, Tesauro M. Association of gut hormones and microbiota with vascular dysfunction in obesity. Nutrients. 2021; 13(2): 613. https://doi.org/10.3390/nu13020613
  • Settanni CR, Ianiro G, Bibbò S, Cammarota G, Gasbarrini A. Gut microbiota alteration and modulation in psychiatric disorders: Current evidence on fecal microbiota transplantation. Prog Neuropsychopharmacol Biol Psychiatry. 2021; 109: 110258. https://doi.org/10.1016/j.pnpbp.2021.110258
  • Smith PA. Brain, meet gut. Nature. 2015; 526: 312-314.
  • Spichak S, Bastiaanssen TFS, Berding K, Vlckova K, Clarke G, Dinan TG, Cryan JF. Mining microbes for mental health: Determining the role of microbial metabolic pathways in human brain health and disease. Neurosci Biobehav Rev. 2021; Mar 3:S0149-7634(21)00103-2. https://doi.org/10.1016/j.neubiorev.2021.02.044
  • Sozbilen GS, Korel F, Yemenicioğlu A. Control of lactic acid bacteria in fermented beverages using lysozyme and nisin: Test of traditional beverage boza as a model food system. Int J Food Sci Tech. 2018; 53(10): 2357-2368. https://doi.org/10.1111/ijfs.13828
  • Todorov SD, Dicks LM. Bacteriocin production by Lactobacillus pentosus ST712BZ isolated from boza. Braz J Microbiol. 2007; 38(1): 166-172. https://doi.org/10.1590/S1517-83822007000100034
  • Troyer EA, Kohn JN, Ecklu-Mensah G, Aleti G, Rosenberg DR, Hong S. Searching for host immune-microbiome mechanisms in obsessive-compulsive disorder: A narrative literature review and future directions. Neurosci Biobehav Rev. 2021; Feb 24:S0149-7634(21)00094-4. https://doi.org/10.1016/j.neubiorev.2021.02.034
  • Valledor SJD, Bucheli JEV, Holzapfel WH, Todorov SD. Exploring beneficial properties of the bacteriocinogenic Enterococcus faecium ST10Bz strain isolated from Boza, a Bulgarian cereal-based beverage. Microorganisms. 2020; 8: 1474. https://doi.org/10.3390/microorganisms8101474
  • Wilkins AT, Reimer RA. Obesity, early life gut microbiota, and antibiotics. Microorganisms. 2021; 9(2): 413. https://doi.org/10.3390/microorganisms9020413

Alteration of Boza Microbiota in the Fermentation Process

Yıl 2021, Cilt: 14 Sayı: 2, 238 - 246, 30.06.2021
https://doi.org/10.30607/kvj.895295

Öz

Boza is a fermented beverage containing beneficial microorganisms for human health. In our study, microbiota present in raw materials used boza production (corn flour, wheat flour, and mayşe), 1st day, 3rd day of boza fermentation and 4th day final product of boza, has been identified by Next Generation DNA Sequencing and metagenomic analysis. As a result of genus-level analysis directly from corn flour and wheat flour samples contained dominantly Streptophyta and Pleomorphobacterium, while in the 1st day, 3rd day, the final product of boza and boza ferment the dominant bacteria were Leuconostoc and Lactococcus at genus level. In the analysis of the pre-enriched samples, the dominant bacteria in corn flour were Enterococcus, Klebsiella, and Micromonospora and in wheat flour were Pantoea and Bacillus. Boza ferment, boza on the 1st day, boza on the 3rd day and the final product of boza dominantly contained Lactococcus. The bacterial diversity, similarity and differences among samples were analyzed by Principal Coordinate Analysis and dendrogram construction. The contribution of raw materials used in the production of boza change to the products at the fermentation stage and to the microbiota during the fermentation process and their contribution to the final product were determined by metagenomic analysis at DNA level.

Kaynakça

  • Altay F, Karbancıoglu-Güler F, Daskaya-Dikmen C, Heperkan D. A review on traditional Turkish fermented non-alcoholic beverages: Microbiota, fermentation process and quality characteristics. Int J Food Microbiol. 2013; 167(1): 44-56. https://doi.org/10.1016/j.ijfoodmicro.2013.06.016
  • Arici M, Daglioglu O. Boza: A lactic acid fermented cereal beverage as a traditional Turkish food. Food Rev Int. 2002; 18(1): 39-48. https://doi.org/10.1081/FRI-120003416
  • Baschali A, Tsakalidou E, Kyriacou A, Karavasiloglou N, Matalas AL. Traditional low-alcoholic and non-alcoholic fermented beverages consumed in European countries: A neglected food group. Nutr Res Rev. 2017; 30(1): 1-24. https://doi.org/10.1017/S0954422416000202
  • Bayat G, Yıldız G. The special fermented Turkish drink: Boza. Journal of Tourism and Gastronomy Studies 2019; 7(4): 2438-2446. https://doi.org/10.21325/jotags.2019.480
  • Bliss ES, Whiteside E. The gut-brain axis, the human gut microbiota and their integration in the development of obesity. Fron Physiol. 2018; 9: 900. https://doi.org/10.3389/fphys.2018.00900
  • Borcaklı M, Öztürk T, Yeşilada E. Cereal source amd microbial consortia of the starter culture influence the chemical composition and physicochemical characteristics of boza. Turk J Agric For. 2018; 42: 412-422. https://doi.org/10.3906/tar-1802-3
  • Botes A, Todorov SD, von Mollendorff JW, Botha A, Dicks LM. Identification of LAB and yeast from boza. Process Biochem. 2007; 42(2): 267-270. https://doi.org/10.1016/j.procbio.2006.07.015
  • Cholakov R, Tumbarski Y, Yanakieva V, Dobrev I, Salim Y, Denkova Z. Antimicrobial acitivity of Leuconostoc lactis strain BT17, isolated from a spontaneously fermented cereal beverage (Boza). J Microbiol Biotech Food Sci. 2017; 7(1): 47-49. https://doi.org/10.15414/jmbfs.2017.7.1.47-49
  • Çakır C. Tarihi, Etimolojisi ve Edebiyatı ile Boza Kitabı, Türk Dünyası Araştırmaları Vakfı Yayınevi, 1. Baskı, İstanbul. 2019; pp. 17-19.
  • De Vuyst L, Leroy F. Functional role of yeasts, lactic acid bacteria, and acetic acid bacteria in cocoa fermentation processes. FEMS Microbiol Rev. 2020; 44(4): 432-453. https://doi.org/10.1093/femsre/fuaa014
  • Dogan M, Ozpinar H. Investigation of probiotic features of bacteria isolated from some food products. Kafkas Univ Vet Fak Derg. 2017; 23(4): 555-562. https://doi.org/10.9775/kvfd.2016.17273
  • Doğan M, Tekiner İH. Extracellular phytase activites of lactic acid bacteria in sourdough mix prepared from traditionally produced boza as starter culture. Food and Health. 2020; 6(2): 117-127. https://doi.org/10.3153/FH20013
  • Gibson GR, Hutkins R, Sanders ME, Prescott SL, Reimer RA, Salminen SJ, Scott K, Stanton C, Swanson KS, Cani PD, Verbeke K, Reid G. The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat Rev Gastroenterol Hepatol. 2017; 14(8): 491-502. https://doi.org/10.1038/nrgastro.2017.75
  • Gotcheva V, Pandiella SS, Angelov A, Roshkova ZG, Webb C. Microflora identification of the Bulgarian cereal-based fermented beverage boza. Process Biochem. 2000; 36(1-2): 127-130. https://doi.org/10.1016/S0032-9592(00)00192-8
  • Gotcheva V, Pandiella SS, Angelov A, Roshkova Z, Webb C. Monitoring the fermentation of the traditional Bulgarian beverage boza. Int J Food Sci Tech. 2001; 36: 129-134. https://doi.org/10.1046/j.1365-2621.2001.00429.x
  • Hancioğlu Ö, Karapinar M. Microflora of Boza, a traditional fermented Turkish beverage. Int J Food Microbiol. 1997; 35(3): 271-274. https://doi.org/10.1016/S0168-1605(96)01230-5
  • Kıvanç M, Yılmaz M, Çakır E. Isolation and identification of lactic acid bacteria from boza, and their microbial activity against several reporter strains. Turk J Biol. 2011; 35(3): 313-324. https://doi.org/10.3906/biy-0906-67
  • Levent H, Cavuldak ÖA. Geleneksel Fermente Bir İçecek: Boza. Akademik Gıda. 2017; 15(3): 300-307. https://doi.org/10.24323/akademik-gida.345273
  • Lindgren SW, Dobrogosz WJ. Antagonistic activities of lactic acid bacteria in food and feed fermentations. FEMS Microbiol Rev. 1990; 87: 149-164. https://doi.org/10.1111/j.1574-6968.1990.tb04885.x
  • Liu D, Ainsworth AJ, Austin FW, Lawrence ML. Use of PCR primers derived from a putative transcriptional regulator gene for species-specific determination of Listeria monocytogenes. Int J Food Microbiol. 2004; 91(3): 297-304. https://doi.org/10.1016/j.ijfoodmicro.2003.07.004
  • Marco ML, Sanders ME, Gänzle M, Arrieta MC, Cotter PD, De Vuyst L, Hill C, Holzapfel W, Lebeer S, Merenstein D, Reid G, Wolfe BE, Hutkins R. The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on fermented foods. Nat Rev Gastroenterol Hepatol. 2021. https://doi.org/10.1038/s41575-020-00390-5
  • Osimani A, Garofalo C, Aquilant L, Milanović V, Clementi F. Unpasteurised commercial boza as a source of microbial diversity. Int J Food Microbiol. 2015; 194: 62-70. http://dx.doi.org/10.1016/j.ijfoodmicro.2014.11.011
  • Palamutoğlu Mİ, Baş M. Traditional Fermented Foods of Turkey. J Health Sci Res. 2020; 2(3): 200-220. https://doi.org/10.46413/boneyusbad.790343
  • Rovella V, Rodia G, Di Daniele F, Cardillo C, Campia U, Noce A, Candi E, Della-Morte D, Tesauro M. Association of gut hormones and microbiota with vascular dysfunction in obesity. Nutrients. 2021; 13(2): 613. https://doi.org/10.3390/nu13020613
  • Settanni CR, Ianiro G, Bibbò S, Cammarota G, Gasbarrini A. Gut microbiota alteration and modulation in psychiatric disorders: Current evidence on fecal microbiota transplantation. Prog Neuropsychopharmacol Biol Psychiatry. 2021; 109: 110258. https://doi.org/10.1016/j.pnpbp.2021.110258
  • Smith PA. Brain, meet gut. Nature. 2015; 526: 312-314.
  • Spichak S, Bastiaanssen TFS, Berding K, Vlckova K, Clarke G, Dinan TG, Cryan JF. Mining microbes for mental health: Determining the role of microbial metabolic pathways in human brain health and disease. Neurosci Biobehav Rev. 2021; Mar 3:S0149-7634(21)00103-2. https://doi.org/10.1016/j.neubiorev.2021.02.044
  • Sozbilen GS, Korel F, Yemenicioğlu A. Control of lactic acid bacteria in fermented beverages using lysozyme and nisin: Test of traditional beverage boza as a model food system. Int J Food Sci Tech. 2018; 53(10): 2357-2368. https://doi.org/10.1111/ijfs.13828
  • Todorov SD, Dicks LM. Bacteriocin production by Lactobacillus pentosus ST712BZ isolated from boza. Braz J Microbiol. 2007; 38(1): 166-172. https://doi.org/10.1590/S1517-83822007000100034
  • Troyer EA, Kohn JN, Ecklu-Mensah G, Aleti G, Rosenberg DR, Hong S. Searching for host immune-microbiome mechanisms in obsessive-compulsive disorder: A narrative literature review and future directions. Neurosci Biobehav Rev. 2021; Feb 24:S0149-7634(21)00094-4. https://doi.org/10.1016/j.neubiorev.2021.02.034
  • Valledor SJD, Bucheli JEV, Holzapfel WH, Todorov SD. Exploring beneficial properties of the bacteriocinogenic Enterococcus faecium ST10Bz strain isolated from Boza, a Bulgarian cereal-based beverage. Microorganisms. 2020; 8: 1474. https://doi.org/10.3390/microorganisms8101474
  • Wilkins AT, Reimer RA. Obesity, early life gut microbiota, and antibiotics. Microorganisms. 2021; 9(2): 413. https://doi.org/10.3390/microorganisms9020413
Toplam 32 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Veteriner Cerrahi
Bölüm ARAŞTIRMA MAKALESİ
Yazarlar

Murat Kavruk Bu kişi benim 0000-0001-5331-7253

Mediha Nur Zafer Yurt 0000-0002-3064-3811

Behiye Büşra Taşbaşı 0000-0002-2076-3756

Elif Esma Acar 0000-0002-6264-7550

Ali Soyuçok 0000-0003-2626-5827

Osman Altunbaş 0000-0003-1840-628X

Veli Cengiz Özalp 0000-0002-7659-5990

Mert Sudağıdan 0000-0002-3980-8344

Yayımlanma Tarihi 30 Haziran 2021
Kabul Tarihi 6 Mayıs 2021
Yayımlandığı Sayı Yıl 2021 Cilt: 14 Sayı: 2

Kaynak Göster

APA Kavruk, M., Yurt, M. N. Z., Taşbaşı, B. B., Acar, E. E., vd. (2021). Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi. Kocatepe Veterinary Journal, 14(2), 238-246. https://doi.org/10.30607/kvj.895295
AMA Kavruk M, Yurt MNZ, Taşbaşı BB, Acar EE, Soyuçok A, Altunbaş O, Özalp VC, Sudağıdan M. Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi. kvj. Haziran 2021;14(2):238-246. doi:10.30607/kvj.895295
Chicago Kavruk, Murat, Mediha Nur Zafer Yurt, Behiye Büşra Taşbaşı, Elif Esma Acar, Ali Soyuçok, Osman Altunbaş, Veli Cengiz Özalp, ve Mert Sudağıdan. “Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi”. Kocatepe Veterinary Journal 14, sy. 2 (Haziran 2021): 238-46. https://doi.org/10.30607/kvj.895295.
EndNote Kavruk M, Yurt MNZ, Taşbaşı BB, Acar EE, Soyuçok A, Altunbaş O, Özalp VC, Sudağıdan M (01 Haziran 2021) Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi. Kocatepe Veterinary Journal 14 2 238–246.
IEEE M. Kavruk, M. N. Z. Yurt, B. B. Taşbaşı, E. E. Acar, A. Soyuçok, O. Altunbaş, V. C. Özalp, ve M. Sudağıdan, “Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi”, kvj, c. 14, sy. 2, ss. 238–246, 2021, doi: 10.30607/kvj.895295.
ISNAD Kavruk, Murat vd. “Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi”. Kocatepe Veterinary Journal 14/2 (Haziran 2021), 238-246. https://doi.org/10.30607/kvj.895295.
JAMA Kavruk M, Yurt MNZ, Taşbaşı BB, Acar EE, Soyuçok A, Altunbaş O, Özalp VC, Sudağıdan M. Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi. kvj. 2021;14:238–246.
MLA Kavruk, Murat vd. “Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi”. Kocatepe Veterinary Journal, c. 14, sy. 2, 2021, ss. 238-46, doi:10.30607/kvj.895295.
Vancouver Kavruk M, Yurt MNZ, Taşbaşı BB, Acar EE, Soyuçok A, Altunbaş O, Özalp VC, Sudağıdan M. Boza Mikrobiyotasının Fermantasyon Sürecindeki Değişimi. kvj. 2021;14(2):238-46.

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