Öz
Bu çalışmada, geleneksel olarak üretilen susuz lahana turşusundan izole edilen laktik asit bakterilerinin (LAB) probiyotik özellikleri incelenmiştir. Bu amaçla, İzmir ili çevresinde geleneksel yöntemlerle üretilen susuz lahana turşusu örneklerinden toplam 39 adet LAB izole edilmiştir. İzolatlara probiyotik özelliklerinin belirlenmesi amacıyla farklı pH, sıcaklık ve tuz değerlerinde gelişim, safra tuzuna tolerans, %0.4 (v/v) fenole dayanıklılık, pepsin ve pankreatin varlığında gelişim, antimikrobiyal aktivite, antibiyotiklere karşı direnç, hemolitik aktivite ve β-Galaktosidaz aktivitesi analizleri uygulanmıştır. Çalışma kapsamında, tüm antibiyotiklere karşı direnç gösterdiği tespit edilen 4 adet izolat dışında (HL7, HL13, HL22 ve HL31), diğer tüm izolatların probiyotik olarak değerlendirilebileceği, ancak bu izolatlar içerisinden GL1, GL3, GL13, HL8, HL9, HL10, HL11, HL12, HL14, HL18, HL25, HL26, HL28, HL29, HL33 kodlu izolatların probiyotik potansiyellerinin, diğer izolatlardan düşük pH ve yüksek NaCl konsantrasyonunda gelişim, safra tuzu, fenol, pepsin ve pankreatin varlığında gelişim, geniş antimikrobiyal spektrum, yüksek antimikrobiyal etki, antibiyotiklere karşı duyarlılık ve β- galaktosidaz aktivitesi açısından değerlendirildiğinde daha yüksek olduğu tespit edilmiştir. Elde edilen sonuçlar, susuz lahana turşusunun önemli bir probiyotik kaynağı olduğunu göstermiştir.
Anahtar Kelimeler
sauerkraut turşu probiyotik laktik asit bakterisi fermente sebze
Destekleyen Kurum
Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü (BAP)
Proje Numarası
FYL-2018-20141
Teşekkür
Bu çalışma, Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü (BAP) tarafından desteklenmiştir (Proje Kodu: FYL-2018-20141).
Kaynakça
- FAO/WHO. (2001). Evaluation of health and nutritional properties of powder milk and live lactic acid bacteria, Food and Agriculture Organization of the United Nations and World Health Organization Expert Consultation Report. www.fao.org/documents/pub_dett.asp?lang=en&pub_id=61756 (Erişim tarihi 2 Mart 2019).
- Kandylis, P., Pissaridi, K., Bekatorou, A., Kanellaki, M., & Koutinas, A. A. (2016). Dairy and non-dairy probiotic beverages. Curr. Opin. Food Sci., 7, 58-63. https://doi.org/10.1016/j.cofs.2015.11.012
- Gibson, G. R., Hutkins, R., Sanders, M. E., Prescott S. L., Reimer, R. A., Salminen, S. J., … & Reid, G. (2017). Expert consensus document: The international scientific association for probiotics and prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat. Rev. Gastroenterol. Hepatol., 4(8), 491-502. https://doi.org/10.1038/nrgastro.2017.75
- Şengün, İ. Y. (2011). Lactic acid bacteria used in the production of fermented foods. Biological Diversity and Conservation, 4(1), 42-53.
- Başdoğan, M. G. (2020). Geleneksel olarak üretilen turşulardan izole edilen laktik asit bakterilerinin probiyotik potansiyelinin belirlenmesi. (Yüksek Lisans Tezi, Ege Üniversitesi, İzmir, Türkiye). Alınan yer Ulusal Tez Merkezi.
- Wang, C. Y., Lin, P. R., Ng, C. C., & Shyu, Y. T. (2010). Probiotic properties of Lactobacillus strains isolated from the feces of breast-fed infants and Taiwanese pickled cabbage. Anaerobe, 16(6), 578–585. https://doi.org/10.1016/j.anaerobe.2010.10.003
- Bhanwar, S., Singh, A., & Ganguli, A. (2014). Probiotic characterization of potential hydrolases producing Lactococcus lactis subsp. lactis isolated from pickled yam. Int. J. Food Sci. Nutr., 65(1), 53–61. https://doi.org/10.3109/09637486.2013.832175
- Tokatlı, M., Gülgör, G., Baʇder Elmacı, S., Arslankoz Işleyen, N. & Özçelik, F. (2015). In vitro properties of potential probiotic indigenous lactic acid bacteria originating from traditional pickles. BioMed Res. Int., 315819. https://doi.org/10.1155/2015/315819
- Zielińska, D., Rzepkowska, A., Radawska, A., & Zieliński, K. (2015). In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber. Current Microbiol., 70(2), 183–194. https://doi.org/10.1007/s00284-014-0699-0
- Monika, S., Kumar, V., Kumari, A., Angmo, K., & Bhalla, T.C. (2017). Isolation and characterization of lactic acid bacteria from traditional pickles of Himachal Pradesh, India. J. Food Sci. Technol., 54(7), 1945–1952. https://doi.org/10.1007/s13197-017-2629-1
- Špička, J., Kalač, P., Bover-Cid, S., & Křížek, M. (2002). Application of lactic acid bacteria starter cultures for decreasing the biogenic amine levels in sauerkraut. Eur. Food Res. Technol., 215(6), 509-514. https://doi.org/10.1007/s00217-002-0590-2
- Mattila, P., & Hellström, J. (2007). Phenolic acids in potatoes, vegetables and some of their products. J. Food Compost. Anal., 20(3-4), 152-160. https://doi.org/10.1016/j.jfca.2006.05.007
- Maden, B. (2020). Alman tipi lahana turşusu fermantasyonu sırasında bazı insektisitlerin değişiminin incelenmesi. (Yüksek Lisans tezi, Bursa Uludağ Üniversitesi, Bursa, Türkiye). Alınan yer https://acikerisim.uludag.edu.tr/handle/11452/15279.
- Beganović, J., Kos, B., Pavunc, A. L., Uroić, K., Jokić, M., & Šušković, J. (2014). Traditionally produced sauerkraut as source of autochthonous functional starter cultures. Microbiol. Res., 169(7-8), 623-632. https://doi.org/10.1016/j.micres.2013.09.015
- Orgeron II, R.P., Corbin, A., & Scott, B. (2016). Sauerkraut: A Probiotic Superfood. Funct.l Foods Health Dis., 6(8), 536-543. https://doi.org/10.31989/ffhd.v6i8.262
- Touret, T., Oliveira, M., & Semedo-Lemsaddek, T. (2018.) Putative probiotic lactic acid bacteria isolated from sauerkraut fermentations. PloS one, 13(9). https://doi.org/10.1371/journal.pone.0203501
- Bulut, Ç. (2003). Isolation and molecular characterization of lactic acid bacteria from cheese. (Master dissertations, University of İzmir Institute of Technology). Alınan yer https://gcris.iyte.edu.tr/handle/11147/3114
- Rohde, R. (1976). Laboratory methods in food and dairy microbiology. In W.F Harrigan & M.E. McCance (Eds.), Journal of basic microbiology (Revised ed., pp 19-20). London: Academic Press.
- Papamanoli, E., Tzanestakis, N., Litopoulou-Tzanetaki, E., & Kotzekidou, P. (2003). Characterisation of lactic acid bacteria isolated from a greek dry- sausages in respect of their technological and probiotic properties. Meat Sci., 65, 859-867. https://doi.org/10.1016/S0309-1740(02)00292-9
- G-Allegria, E., Lopez, I., Ruiz, I. J., Saenz, J., Fernandez, E., Zarazaga, M., .... & Ruiz-Larrea, F. (2004). High tolerance of wild Lactobacillus plantarum and Oenococcus oeni strains to lyophilisation and stress environmental conditions of acid pH and ethanol. FEMS Microbiol. Lett., 230, 53-61. https://doi.org/10.1016/S0378-1097(03)00854-1
- Hébert, E. M., Raya, R. R., Tailliez, P., & De Giori, G.S. (2000). Characterization of natural Isolates of Lactobacillus strains to be used as starter cultures in dairy fermentation. Int. J. Food Microbiol., 59, 19-27. https://doi.org/10.1016/S0168-1605(00)00282-8
- Adnan, A . F. M., & Tan, I. K. (2007). Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential. Bioresour. Technol., 98, 1380-1385. https://doi.org/10.1016/j.biortech.2006.05.034
- Musikasang, H., Tani, A., H-kittikun, A., & Maneerat, S. (2009). Probiotic potential of lactic acid bacteria isolated from chicken gastrointestinal digestive tract World J. Microbiol. Biotechnol., 25, 1337-1345. https://doi.org/10.1007/s11274-009-0020-8
- Wootton, M. (2013). BSAC methods for antimicrobial susceptibility testing. The British Society for Antimicrobial Chemotherapy, 44, 1–87.
- Tambekar, D. H., & Bhutada, S. A. (2010). An evaluation of probiotic potential of lactobacillus sp. from milk of domestic animals and commercial available probiotic preparations in prevention of enteric bacterial infections. Recent Res. Sci. Technol., 2, 82-88.
- Boricha, A. A., Shekh, S. L., Pithva, S. P., Ambalam, P. S., & Vyas, B. R. M. (2019). In vitro evaluation of probiotic properties of Lactobacillus species of food and human origin. LWT-Food Sci. Technol., 106, 201-208. https://doi.org/10.1016/j.lwt.2019.02.021
- Şimşek, Ö. (2003). Uşak ve yöresi ekşi hamurlarından izole edilen antimikrobiyal aktiviteye sahip laktik asit bakterilerinin tanımlanması ve bazı metabolik özelliklerinin belirlenmesi. (Yüksek Lisans Tezi, Pamukkale Üniversitesi, Denizli, Türkiye). Alınan yer http://acikerisim.pau.edu.tr/handle
- Ragul, K., Syiem, I., Sundar, K., & Shetty, P. H. (2017). Characterization of probiotic potential of Bacillus species isolated from a traditional brine pickle. J. Food Sci. Technol., 54 (13), 4483. https://doi.org/10.1007/s13197-017-2928-6
- Rao, Y., Tao, Y., Li, Y., She, X., Yang, J., Qian Y., ... & Xia, H. (2019). Characterization of a probiotic starter culture with anti-Candida activity for Chinese pickle fermentation. Food Funct., 10, 6936. https://doi.org/10.1039/C9FO01191A
- Xanthopoulos, V., Litopoulou-Tzanetaki, E., & Tzanetaki, N. (2000). Characterization of Lactobacillus isolates frominfant faeces as dietary adjuncts. Food Microbiol., 17, 205-215. https://doi.org/10.1006/fmic.1999.0300
- Cizeikiene, D., Juodeikiene, G., Paskevicius, A., & Bartkiene, E. (2013). Antimicrobial activity of lactic acid bacteria against pathogenic and spoilage microorganism isolated from food and their control in wheat bread, Food Control, 31(2), 539–545. https://doi.org/10.1016/j.foodcont.2012.12.004
- Sharaf, E. F., & Al Harbi, R. M. (2011). Isolation, identification and antimicrobial activity of some local isolates of lactic acid bacteria. Res. J. Microbiol., 6(12), 826-838.
- Ashraf, R., & Shah, N. P. (2011). Antibiotic resistance of probiotic organisms and safety of probiotic dairy products. Int. Food Res. J., 18(3), 837-853.
- Maragkoudakis, P. A., Zoumpopoulor, G., Miaris, C., Kalantzopoulos. G., Pot, B., & Tsakalidou, E. (2006). Probiotic potential of Lactobacillus strains isolated from dairy products. Int. Dairy J., 16, 189–199. https://doi.org/10.1016/j.idairyj.2005.02.009
- Harrington, L. K., & Mayberry, J. F. (2008). A re‐appraisal of lactose intolerance. Int. J. Clin. Pract., 62(10), 1541-1546. https://doi.org/10.1111/j.1742-1241.2008.01834.x
Öz
The present study investigated the probiotic properties of lactic acid bacteria (LAB) isolated from traditionally produced sauerkraut. For this reason, total of thirty-nine isolates were obtained from sauerkraut produced at homes by traditional methods around Izmir city. Development of different pH, temperature and salt values, tolerance to bile salt, resistance to 0.4% (v/v) phenol, development in the presence of pepsin and pancreatin, antimicrobial activity, resistance to antibiotics, hemolytic activity and β-Galactosidase activity analyzes were applied to the isolates for determining probiotic properties. Within the scope of the study, except for four isolates (HL7, HL13, HL22 and HL31), which were found to be resistant to all antibiotics, all other isolates could be considered as probiotics. However, among these isolates, the probiotic potential of GL1, GL3, GL13, HL8, HL9, HL10, HL11, HL12, HL14 , HL18, HL25, HL26, HL28, HL29, HL33 was found higher than other isolates when investigated in terms of growth at low pH and high NaCl concentration, growth in the presence of bile salt, phenol, pepsin and pancreatin, broad antimicrobial spectrum, high antimicrobial effect, sensitivity to antibiotics and β-galactosidase activity. The results showed that sauerkraut is an important probiotic source.
Anahtar Kelimeler
sauerkraut pickle lactic acid bacteria fermented vegetables probiotics
Proje Numarası
FYL-2018-20141
Kaynakça
- FAO/WHO. (2001). Evaluation of health and nutritional properties of powder milk and live lactic acid bacteria, Food and Agriculture Organization of the United Nations and World Health Organization Expert Consultation Report. www.fao.org/documents/pub_dett.asp?lang=en&pub_id=61756 (Erişim tarihi 2 Mart 2019).
- Kandylis, P., Pissaridi, K., Bekatorou, A., Kanellaki, M., & Koutinas, A. A. (2016). Dairy and non-dairy probiotic beverages. Curr. Opin. Food Sci., 7, 58-63. https://doi.org/10.1016/j.cofs.2015.11.012
- Gibson, G. R., Hutkins, R., Sanders, M. E., Prescott S. L., Reimer, R. A., Salminen, S. J., … & Reid, G. (2017). Expert consensus document: The international scientific association for probiotics and prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat. Rev. Gastroenterol. Hepatol., 4(8), 491-502. https://doi.org/10.1038/nrgastro.2017.75
- Şengün, İ. Y. (2011). Lactic acid bacteria used in the production of fermented foods. Biological Diversity and Conservation, 4(1), 42-53.
- Başdoğan, M. G. (2020). Geleneksel olarak üretilen turşulardan izole edilen laktik asit bakterilerinin probiyotik potansiyelinin belirlenmesi. (Yüksek Lisans Tezi, Ege Üniversitesi, İzmir, Türkiye). Alınan yer Ulusal Tez Merkezi.
- Wang, C. Y., Lin, P. R., Ng, C. C., & Shyu, Y. T. (2010). Probiotic properties of Lactobacillus strains isolated from the feces of breast-fed infants and Taiwanese pickled cabbage. Anaerobe, 16(6), 578–585. https://doi.org/10.1016/j.anaerobe.2010.10.003
- Bhanwar, S., Singh, A., & Ganguli, A. (2014). Probiotic characterization of potential hydrolases producing Lactococcus lactis subsp. lactis isolated from pickled yam. Int. J. Food Sci. Nutr., 65(1), 53–61. https://doi.org/10.3109/09637486.2013.832175
- Tokatlı, M., Gülgör, G., Baʇder Elmacı, S., Arslankoz Işleyen, N. & Özçelik, F. (2015). In vitro properties of potential probiotic indigenous lactic acid bacteria originating from traditional pickles. BioMed Res. Int., 315819. https://doi.org/10.1155/2015/315819
- Zielińska, D., Rzepkowska, A., Radawska, A., & Zieliński, K. (2015). In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber. Current Microbiol., 70(2), 183–194. https://doi.org/10.1007/s00284-014-0699-0
- Monika, S., Kumar, V., Kumari, A., Angmo, K., & Bhalla, T.C. (2017). Isolation and characterization of lactic acid bacteria from traditional pickles of Himachal Pradesh, India. J. Food Sci. Technol., 54(7), 1945–1952. https://doi.org/10.1007/s13197-017-2629-1
- Špička, J., Kalač, P., Bover-Cid, S., & Křížek, M. (2002). Application of lactic acid bacteria starter cultures for decreasing the biogenic amine levels in sauerkraut. Eur. Food Res. Technol., 215(6), 509-514. https://doi.org/10.1007/s00217-002-0590-2
- Mattila, P., & Hellström, J. (2007). Phenolic acids in potatoes, vegetables and some of their products. J. Food Compost. Anal., 20(3-4), 152-160. https://doi.org/10.1016/j.jfca.2006.05.007
- Maden, B. (2020). Alman tipi lahana turşusu fermantasyonu sırasında bazı insektisitlerin değişiminin incelenmesi. (Yüksek Lisans tezi, Bursa Uludağ Üniversitesi, Bursa, Türkiye). Alınan yer https://acikerisim.uludag.edu.tr/handle/11452/15279.
- Beganović, J., Kos, B., Pavunc, A. L., Uroić, K., Jokić, M., & Šušković, J. (2014). Traditionally produced sauerkraut as source of autochthonous functional starter cultures. Microbiol. Res., 169(7-8), 623-632. https://doi.org/10.1016/j.micres.2013.09.015
- Orgeron II, R.P., Corbin, A., & Scott, B. (2016). Sauerkraut: A Probiotic Superfood. Funct.l Foods Health Dis., 6(8), 536-543. https://doi.org/10.31989/ffhd.v6i8.262
- Touret, T., Oliveira, M., & Semedo-Lemsaddek, T. (2018.) Putative probiotic lactic acid bacteria isolated from sauerkraut fermentations. PloS one, 13(9). https://doi.org/10.1371/journal.pone.0203501
- Bulut, Ç. (2003). Isolation and molecular characterization of lactic acid bacteria from cheese. (Master dissertations, University of İzmir Institute of Technology). Alınan yer https://gcris.iyte.edu.tr/handle/11147/3114
- Rohde, R. (1976). Laboratory methods in food and dairy microbiology. In W.F Harrigan & M.E. McCance (Eds.), Journal of basic microbiology (Revised ed., pp 19-20). London: Academic Press.
- Papamanoli, E., Tzanestakis, N., Litopoulou-Tzanetaki, E., & Kotzekidou, P. (2003). Characterisation of lactic acid bacteria isolated from a greek dry- sausages in respect of their technological and probiotic properties. Meat Sci., 65, 859-867. https://doi.org/10.1016/S0309-1740(02)00292-9
- G-Allegria, E., Lopez, I., Ruiz, I. J., Saenz, J., Fernandez, E., Zarazaga, M., .... & Ruiz-Larrea, F. (2004). High tolerance of wild Lactobacillus plantarum and Oenococcus oeni strains to lyophilisation and stress environmental conditions of acid pH and ethanol. FEMS Microbiol. Lett., 230, 53-61. https://doi.org/10.1016/S0378-1097(03)00854-1
- Hébert, E. M., Raya, R. R., Tailliez, P., & De Giori, G.S. (2000). Characterization of natural Isolates of Lactobacillus strains to be used as starter cultures in dairy fermentation. Int. J. Food Microbiol., 59, 19-27. https://doi.org/10.1016/S0168-1605(00)00282-8
- Adnan, A . F. M., & Tan, I. K. (2007). Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential. Bioresour. Technol., 98, 1380-1385. https://doi.org/10.1016/j.biortech.2006.05.034
- Musikasang, H., Tani, A., H-kittikun, A., & Maneerat, S. (2009). Probiotic potential of lactic acid bacteria isolated from chicken gastrointestinal digestive tract World J. Microbiol. Biotechnol., 25, 1337-1345. https://doi.org/10.1007/s11274-009-0020-8
- Wootton, M. (2013). BSAC methods for antimicrobial susceptibility testing. The British Society for Antimicrobial Chemotherapy, 44, 1–87.
- Tambekar, D. H., & Bhutada, S. A. (2010). An evaluation of probiotic potential of lactobacillus sp. from milk of domestic animals and commercial available probiotic preparations in prevention of enteric bacterial infections. Recent Res. Sci. Technol., 2, 82-88.
- Boricha, A. A., Shekh, S. L., Pithva, S. P., Ambalam, P. S., & Vyas, B. R. M. (2019). In vitro evaluation of probiotic properties of Lactobacillus species of food and human origin. LWT-Food Sci. Technol., 106, 201-208. https://doi.org/10.1016/j.lwt.2019.02.021
- Şimşek, Ö. (2003). Uşak ve yöresi ekşi hamurlarından izole edilen antimikrobiyal aktiviteye sahip laktik asit bakterilerinin tanımlanması ve bazı metabolik özelliklerinin belirlenmesi. (Yüksek Lisans Tezi, Pamukkale Üniversitesi, Denizli, Türkiye). Alınan yer http://acikerisim.pau.edu.tr/handle
- Ragul, K., Syiem, I., Sundar, K., & Shetty, P. H. (2017). Characterization of probiotic potential of Bacillus species isolated from a traditional brine pickle. J. Food Sci. Technol., 54 (13), 4483. https://doi.org/10.1007/s13197-017-2928-6
- Rao, Y., Tao, Y., Li, Y., She, X., Yang, J., Qian Y., ... & Xia, H. (2019). Characterization of a probiotic starter culture with anti-Candida activity for Chinese pickle fermentation. Food Funct., 10, 6936. https://doi.org/10.1039/C9FO01191A
- Xanthopoulos, V., Litopoulou-Tzanetaki, E., & Tzanetaki, N. (2000). Characterization of Lactobacillus isolates frominfant faeces as dietary adjuncts. Food Microbiol., 17, 205-215. https://doi.org/10.1006/fmic.1999.0300
- Cizeikiene, D., Juodeikiene, G., Paskevicius, A., & Bartkiene, E. (2013). Antimicrobial activity of lactic acid bacteria against pathogenic and spoilage microorganism isolated from food and their control in wheat bread, Food Control, 31(2), 539–545. https://doi.org/10.1016/j.foodcont.2012.12.004
- Sharaf, E. F., & Al Harbi, R. M. (2011). Isolation, identification and antimicrobial activity of some local isolates of lactic acid bacteria. Res. J. Microbiol., 6(12), 826-838.
- Ashraf, R., & Shah, N. P. (2011). Antibiotic resistance of probiotic organisms and safety of probiotic dairy products. Int. Food Res. J., 18(3), 837-853.
- Maragkoudakis, P. A., Zoumpopoulor, G., Miaris, C., Kalantzopoulos. G., Pot, B., & Tsakalidou, E. (2006). Probiotic potential of Lactobacillus strains isolated from dairy products. Int. Dairy J., 16, 189–199. https://doi.org/10.1016/j.idairyj.2005.02.009
- Harrington, L. K., & Mayberry, J. F. (2008). A re‐appraisal of lactose intolerance. Int. J. Clin. Pract., 62(10), 1541-1546. https://doi.org/10.1111/j.1742-1241.2008.01834.x
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Koruma ve Biyolojik Çeşitlilik |
| Bölüm | Research Article |
| Yazarlar | |
| Proje Numarası | FYL-2018-20141 |
| Erken Görünüm Tarihi | 14 Nisan 2022 |
| Yayımlanma Tarihi | 15 Nisan 2022 |
| Gönderilme Tarihi | 8 Aralık 2021 |
| Kabul Tarihi | 15 Ocak 2022 |
| Yayımlandığı Sayı | Yıl 2022 Cilt: 15 Sayı: 1 |
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❖ ISSN 1308-5301 Baskı; ISSN 1308-8084 Çevrimiçi
❖ Yayım Başlangıç Tarihi 2008
❖ © Copyright by Biyolojik Çeşitlilik ve Koruma/Biyolojik Çeşitlilik ve Koruma-www.biodicon.com/ adresinden çevrimiçi olarak erişime açıktır. Tüm hakları saklıdır
. ❖ Yayımcı : ERSİN YÜCEL (https://www.ersinyucel.com.tr/)
❖ Bu dergi yılda üç sayı olarak yayımlanır. Eskişehir/Türkiye'de basılmaktadır.
❖ Bu dergide yayımlanan makalelerin her türlü sorumluluğu yazarlarına aittir.
❖ Editör / Genel Yayın Yönetmeni : Prof.Dr. Ersin YÜCEL, https://orcid.org/0000-0001-8274-7578