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Mogan Gölü’ndeki bazı balık türlerinde vitellogenin proteininin elektroforetik karşılaştırılması

Year 2016, Volume: 33 Issue: 2, 151 - 156, 15.05.2016
https://doi.org/10.12714/egejfas.2016.33.2.09

Abstract

Öz: Bu çalışma, balıkçılık ve rekreasyon alanı olarak kullanılan Mogan Gölü’nde (Ankara - Türkiye) Ocak-Mayıs 2013 ayları arasında yürütülmüştür. Gölde yaşayan balık türlerinden Cyprinus carpio, Carassius gibelio, Tinca tinca ve Esox lucius türlerinin erkek ve dişi bireylerinin kan plazmasında elektroforetik SDS-Page (Sodyum Dodesil Sülfat Poliakrilamid Jel Elektroforezi) metodu ile vitellogenin (VTG) proteini araştırılmış ve türler arası karşılaştırma yapılmıştır. Bu metot için Biometra marka dikey elektroforez cihazı kullanılmıştır. Üreme döneminde C. carpio, T. tinca ve E. Lucius’un erkek bireylerinde vitellogenin proteini saptanmıştır. Göl ortamındaki çeşitli kirleticilerin bu sonuçlar üzerinde etkili olduğu düşünülmektedir. Bu çalışmada elde edilen bulgulara göre kullanılan testin iyi bir biyo-belirteç olduğu anlaşılmıştır.

Anahtar kelimeler: Vitellogenin, elektroforez, Cyprinus carpio, Carassius gibelio, Tinca tinca, Esox lucius

References

  • Allner, B., Wegener, G., Knacker, T. & Stahlschmidt-Allner, P. (1999). Electrophoretic determination of estrogen-induced protein in fish exposed to synthetic and naturally occurring chemicals. The Science of the Total Environment, 233(1-3): 21-31. doi:10.1016/S0048-9697(99)00176-X
  • Asturiano, J.F., Romaguera, F., Aragon, P., Atienza, J., Puchades, R. & Maquieira, A. (2005). Sandwich immunoassay for determination of vitellogenin in golden grey mullet (Liza aurata) serum as a field exposure biomarker. Analytical and Bioanalytical Chemistry, 381: 1152-1160. doi:10.1007/s00216-004-3049-4
  • Bjorkblom, C., Mustamaki, N., Olsson, P., Katsiadaki, I. & Wiklund, T., (2011). Assessment of reproductive biomarkers in three-spined stickleback (Gasterosteus aculeatus) from sewage effluent recipients. Environmental Toxicology, 28: 229–237. doi: 10.1002/tox.20715
  • Brion, F., Rogerieux, F., Noury, P., Migeon, B., Flammarion, P., Thybaud, E. & Porcher, J.M. (2000). Two-step purification method of vitellogenin from three teleost fish species: rainbow trout (Oncorhynchus mykiss), gudgeon (Gobio gobio) and chub (Leuciscus cephalus). Journal of Chromatography. B, Biomedical Sciences and Applications, 737(1-2): 3-12. doi:10.1016/S0378-4347(99)00406-5
  • Colborn, T., vom Saal, F.S. & Soto, A. M. (1993). Developmental effects of endocrine-disrupting chemicals in wildlife and humans. Environmental Health Perspectives, 101(5): 378-384. doi:10.2307/3431890
  • Coldham, N.G., Mehul, D., Sivapathasundaram, S., McDonnell, D.P., Conor, C. & Sauer, M.J. (1997). Evaluation of a recombinant yeast cell estrogen screening assay. Environmental Health Perspectives, 105: 734-742. doi:10.2307/3433729
  • Çek, Ş. & Sarıhan, F. (2010). The Effects of Sex Steroids of Endocrine Disrupting Chemicals on Fishes (in Turkish with English abstract). Ege Journal of Fisheries and Aquatic Sciences, 27(1): 41-46.
  • Donohoe, R.M. & Curtis, L.R. (1996). Estrogenic activity of chlordecone, o,p’-DDT and o,p’-DDE in juvenile rainbow trout: induction of vitellogenesis and interaction with hepatic estrogen binding sites. Aquatic Toxicology, 36: 31-52. doi:10.1016/S0166-445X(96)00799-0
  • Fakıoğlu, Ö. & Polatsü, S. (2005). Determination of external phosphorus loading in Mogan Lake (Ankara) following some restoration measures (in Turkish with English abstract) Yuzuncu Yil University Journal of Agricultural Sciences, 15(1): 63-69.
  • Fenech, M., (2000). The in vitro micronucleus technique. Mutation Research, 455, 81-95. doi:10.1016/S0027-5107(00)00065-8
  • Fenech, M., Holland, N., Chang, W.P., Zeiger, E. & Bonassi, S. (1999). The human micronucleus project-an international collaborative study on the use of micronucleus technique for measuring DNA damage in humans. Mutation Research, 428: 271-283. PII:S1383-57429900053-8.
  • Hennies, M., Wiesmann, M., Allner, B. & Sauerwein, H. (2003). Vitellogenin in carp (Cyprinus carpio) and perch (Perca fluviatilis): purification, characterization and development of an ELISA for the detection of estrogenic effects. The Science of the Total Environment, 309(1-3), 93-103. doi:10.1016/S0048-9697(03)00005-6
  • Jobling, S., Sheanan, D., Osborne, J.A., Matthiesen, P. & Sumpter, J.P. (1996). Inhibition of testicular growth in rainbow trout (Oncorhynchus mykiss) exposed to alkylphenolic chemicals. Environmental Toxicology and Chemistry 15: 194–202. doi:10.1002/etc.5620150218
  • Jobling, S. & Sumpter, J.P. (1993). Detergent components in sewage effluent are weakly estrogenic to fish: an in vitro study using rainbow trout (Oncorhynchus mykiss) hepatocytes. Aquatic Toxicology, 27: 361–372. doi:10.1016/0166-445X(93)90064-8
  • Karul, C., Soyupak, S., Çilesiz, A. F., Akbay, N. & Germen, E. (2000). Case studies on the use of neural networks in eutrophication modeling. Ecological Modelling, 134: 145–152.
  • doi:10.1016/S0304-3800(00)00360-4
  • Keleştemur, G.T. & Özdemir, Y. (2010). Effect of low oxygen level on some blood parameter levels of fry rainbow trouts (Oncorhynchus mykiss, W. 1792). (in Turkish with English abstract). Journal of FisheriesSciences.com, 4(4): 310-317.
  • Lagler, K.F. (1956). Freshwater Fishery Biology. Dubuque, Iowa: WMC. Brown Company. Publications.
  • Lazier, C.B. & McKay, M.E. (1993). Vitellogenin gene expression in teleost fish. In: Hochachka, P.W., Mommsen, T.P. (Eds.), Biochemistry and Molecular Biology of Fishes, Vol. 2. Elsevier, Amsterdam, The Netherlands.
  • Maltais, D. & Roy, R.L. (2009). Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody. Fish Physiology and Biochemistry, 35: 241-250. doi:10.1007/s10695-008-9205-6
  • Meucci, V. & Arukwe, A. (2005). Detection of vitellogenin and zona radiata protein expressions in surface mucus of immature juvenile Atlantic salmon (Salmo salar) exposed to waterborne nonylphenol. Aquatic Toxicology, 73(1): 1-10. PMID:15871908.
  • Nagler, J.J., Ruby, S.M., Idler, D.R. & So, Y.P. (1987). Serum phosphoprotein phosphorous and calcium levels as reproductive indicators of vitellogenin in highly vitellogenic mature female and estradiol-injected immature rainbow trout (Salmo gairdneri). Canadian Journal of Zoology, 65(10): 2421-2425. doi:10.1139/z87-365
  • Ndiaye, P., Forgue, J., Lamothe, V., Cauty, C., Tacon, P., Lafon, P., Davail, B., Fostier, A., Le Menn, F. & Nunez, J. (2006). Tilapia (Oreochromis niloticus) vitellogenins: development of homologous and heterologous ELISAs and analysis of vitellogenin pathway through the ovarian follicle. Journal of Experimental Zoology Part A: Comparative Experimental Biology, 305A(7), 576-593. ISSN: 0022-104X.
  • Roubal, W.T., Lomax, D.P., Willis, M.L. & Johnson, L.L. (1997). Purification and partial characterization of English sole (Pleuronectes vetulus) vitellogenin. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 118(3): 613-622. doi:10.1016/S0305-0491(97)00241-1
  • Routledge, E.J., Sheahan, D., Desbrow, C., Brighty, G.C., Waldock, M. & Sumpter, J.P. (1998). Identification of estrogenic chemicals in STW effluent 2: in vivo responses in trout and roach. Envıronmental Science and Technology, 32: 1559–1565. doi:10.1021/es970796a
  • Schiffmann, D. & De Boni, U. (1991). Dislocation of chromatin elements in prophase induced diethylstilbestrol: a novel mechanism by which micronuclei can arise. Mutation Research, 246(1): 113–122. doi:10.1016/0027-5107(91)90113-3
  • Shore, L., Gurevitz, M. & Shemesh, M. (1993). Estrogen as environmental pollutant. Bulletin of Environmental Contamination and Toxicology, 51: 361–366. doi:10.1007/BF00201753
  • Stumpf, M., Ternes, T.A., Ilaber, K. & Baumann, W. (1996). Determination of natural and synthetic estrogens in sewage plants and river water. Vom Wasser, 87: 251–261.
  • Sumpter, J.P. & Jobling, S. (1995). Vitellogenesis as biomarker for estrogenic contamination of the aquatic environment. Environmental Health Perspective, 103(7): 173–178. doi:10.2307/3432529
  • Ternes, T.A., Stumph, M., Mueller, J., Haberer, K., Wilken, R.D. & Servos, M. (1999). Behavior and occurrence of estrogens in municipal sewage treatment plants. I. Investigations in Germany, Canada and Brazil. Science of The Total Environment, 225: 81–90.
  • doi:10.1016/S0048-9697(98)00334-9
  • Tyler, C.R. & Sumpter, J. P. (1996). Oocyte growth and development in teleosts. Reviews in Fish Biology and Fisheries, 6: 287-318. doi:10.1007/BF00122584
  • Tyler, C.R. & Sumpter, J. P. (1990). The development of a radioimmunoassay for carp (Cyprinus carpio) vitellogenin. Fish Physiology and Biochemistry, 8(2): 129-140. doi:10.1007/BF00004440
  • Tyler, C.R., van der Eerden, B., Jobling, S., Panter, G. & Sumpter, J.P. (1996). Measurement of vitellogenin, a biomarker for exposure to oestrogenic chemicals, in a wide variety of cyprinid fish. Journal of Comparative Physiology B, 166: 418–426. doi:10.1007/BF02337886
  • Van den Belt, K., Verheyen, R. & Witters, H. (2003). Comparison of vitellogenin responses in zebrafish and rainbow trout following exposure to environmental estrogens. Ecotoxicology and Environmental Safety, 56: 271-281. doi:10.1016/S0147-6513(03)00004-6
  • Van den Belt, K., Verheyen, R. & Witters, H. (2001). Reproductive effects of ethynylestradiol and 4t-octylphenol on the zebrafish. Archives of Environmental Contamination and Toxicology, 41: 458–467. doi:10.1007/s002440010272
  • Yamanaka, Y., Mizuno,T., Sasai, Y., Kishi, M., Takeda, H., Kim C.H., Hibi, M. & Hirano, T. (1998). A novel homeobox gene, dharma, can induce the organizer in a non-cell-autonomous manner. Genes and Development, 12(15), 2345–2353. doi:10.1101/gad.12.15.2345
  • Yamaguchi, A., Ishibashi, H., Kohra, S., Arizono, K., Kato, K., Nakahama, T., Kanno, Y., Inouye, Y. & Tominaga, N. (2009). Expression analysis of estrogen-responsive genes vitellogenin 1 and 2 in liver of male medaka (Oryzias latipes) exposed to selective ligands of estrogen receptor subtypes. Journal of Health Science, 55(6): 930-938. doi:10.1248/jhs.55.930

Mogan Gölü’ndeki bazı balık türlerinde vitellogenin proteininin elektroforetik karşılaştırılması

Year 2016, Volume: 33 Issue: 2, 151 - 156, 15.05.2016
https://doi.org/10.12714/egejfas.2016.33.2.09

Abstract

Bu çalışma, balıkçılık ve rekreasyon alanı olarak kullanılan Mogan Gölü’nde (Ankara - Türkiye) Ocak-Mayıs 2013 ayları arasında yürütülmüştür. Gölde yaşayan balık türlerinden Cyprinus carpio, Carassius gibelio, Tinca tinca ve Esox lucius türlerinin erkek ve dişi bireylerinin kan plazmasında elektroforetik SDS-Page (Sodyum Dodesil Sülfat Poliakrilamid Jel Elektroforezi) metodu ile vitellogenin (VTG) proteini araştırılmış ve türler arası karşılaştırma yapılmıştır. Bu metot için Biometra marka dikey elektroforez cihazı kullanılmıştır. Üreme döneminde C. carpio, T. tinca ve E. Lucius’un erkek bireylerinde vitellogenin proteini saptanmıştır. Göl ortamındaki çeşitli kirleticilerin bu sonuçlar üzerinde etkili olduğu düşünülmektedir. Bu çalışmada elde edilen bulgulara göre kullanılan testin iyi bir biyo-belirteç olduğu anlaşılmıştır

References

  • Allner, B., Wegener, G., Knacker, T. & Stahlschmidt-Allner, P. (1999). Electrophoretic determination of estrogen-induced protein in fish exposed to synthetic and naturally occurring chemicals. The Science of the Total Environment, 233(1-3): 21-31. doi:10.1016/S0048-9697(99)00176-X
  • Asturiano, J.F., Romaguera, F., Aragon, P., Atienza, J., Puchades, R. & Maquieira, A. (2005). Sandwich immunoassay for determination of vitellogenin in golden grey mullet (Liza aurata) serum as a field exposure biomarker. Analytical and Bioanalytical Chemistry, 381: 1152-1160. doi:10.1007/s00216-004-3049-4
  • Bjorkblom, C., Mustamaki, N., Olsson, P., Katsiadaki, I. & Wiklund, T., (2011). Assessment of reproductive biomarkers in three-spined stickleback (Gasterosteus aculeatus) from sewage effluent recipients. Environmental Toxicology, 28: 229–237. doi: 10.1002/tox.20715
  • Brion, F., Rogerieux, F., Noury, P., Migeon, B., Flammarion, P., Thybaud, E. & Porcher, J.M. (2000). Two-step purification method of vitellogenin from three teleost fish species: rainbow trout (Oncorhynchus mykiss), gudgeon (Gobio gobio) and chub (Leuciscus cephalus). Journal of Chromatography. B, Biomedical Sciences and Applications, 737(1-2): 3-12. doi:10.1016/S0378-4347(99)00406-5
  • Colborn, T., vom Saal, F.S. & Soto, A. M. (1993). Developmental effects of endocrine-disrupting chemicals in wildlife and humans. Environmental Health Perspectives, 101(5): 378-384. doi:10.2307/3431890
  • Coldham, N.G., Mehul, D., Sivapathasundaram, S., McDonnell, D.P., Conor, C. & Sauer, M.J. (1997). Evaluation of a recombinant yeast cell estrogen screening assay. Environmental Health Perspectives, 105: 734-742. doi:10.2307/3433729
  • Çek, Ş. & Sarıhan, F. (2010). The Effects of Sex Steroids of Endocrine Disrupting Chemicals on Fishes (in Turkish with English abstract). Ege Journal of Fisheries and Aquatic Sciences, 27(1): 41-46.
  • Donohoe, R.M. & Curtis, L.R. (1996). Estrogenic activity of chlordecone, o,p’-DDT and o,p’-DDE in juvenile rainbow trout: induction of vitellogenesis and interaction with hepatic estrogen binding sites. Aquatic Toxicology, 36: 31-52. doi:10.1016/S0166-445X(96)00799-0
  • Fakıoğlu, Ö. & Polatsü, S. (2005). Determination of external phosphorus loading in Mogan Lake (Ankara) following some restoration measures (in Turkish with English abstract) Yuzuncu Yil University Journal of Agricultural Sciences, 15(1): 63-69.
  • Fenech, M., (2000). The in vitro micronucleus technique. Mutation Research, 455, 81-95. doi:10.1016/S0027-5107(00)00065-8
  • Fenech, M., Holland, N., Chang, W.P., Zeiger, E. & Bonassi, S. (1999). The human micronucleus project-an international collaborative study on the use of micronucleus technique for measuring DNA damage in humans. Mutation Research, 428: 271-283. PII:S1383-57429900053-8.
  • Hennies, M., Wiesmann, M., Allner, B. & Sauerwein, H. (2003). Vitellogenin in carp (Cyprinus carpio) and perch (Perca fluviatilis): purification, characterization and development of an ELISA for the detection of estrogenic effects. The Science of the Total Environment, 309(1-3), 93-103. doi:10.1016/S0048-9697(03)00005-6
  • Jobling, S., Sheanan, D., Osborne, J.A., Matthiesen, P. & Sumpter, J.P. (1996). Inhibition of testicular growth in rainbow trout (Oncorhynchus mykiss) exposed to alkylphenolic chemicals. Environmental Toxicology and Chemistry 15: 194–202. doi:10.1002/etc.5620150218
  • Jobling, S. & Sumpter, J.P. (1993). Detergent components in sewage effluent are weakly estrogenic to fish: an in vitro study using rainbow trout (Oncorhynchus mykiss) hepatocytes. Aquatic Toxicology, 27: 361–372. doi:10.1016/0166-445X(93)90064-8
  • Karul, C., Soyupak, S., Çilesiz, A. F., Akbay, N. & Germen, E. (2000). Case studies on the use of neural networks in eutrophication modeling. Ecological Modelling, 134: 145–152.
  • doi:10.1016/S0304-3800(00)00360-4
  • Keleştemur, G.T. & Özdemir, Y. (2010). Effect of low oxygen level on some blood parameter levels of fry rainbow trouts (Oncorhynchus mykiss, W. 1792). (in Turkish with English abstract). Journal of FisheriesSciences.com, 4(4): 310-317.
  • Lagler, K.F. (1956). Freshwater Fishery Biology. Dubuque, Iowa: WMC. Brown Company. Publications.
  • Lazier, C.B. & McKay, M.E. (1993). Vitellogenin gene expression in teleost fish. In: Hochachka, P.W., Mommsen, T.P. (Eds.), Biochemistry and Molecular Biology of Fishes, Vol. 2. Elsevier, Amsterdam, The Netherlands.
  • Maltais, D. & Roy, R.L. (2009). Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody. Fish Physiology and Biochemistry, 35: 241-250. doi:10.1007/s10695-008-9205-6
  • Meucci, V. & Arukwe, A. (2005). Detection of vitellogenin and zona radiata protein expressions in surface mucus of immature juvenile Atlantic salmon (Salmo salar) exposed to waterborne nonylphenol. Aquatic Toxicology, 73(1): 1-10. PMID:15871908.
  • Nagler, J.J., Ruby, S.M., Idler, D.R. & So, Y.P. (1987). Serum phosphoprotein phosphorous and calcium levels as reproductive indicators of vitellogenin in highly vitellogenic mature female and estradiol-injected immature rainbow trout (Salmo gairdneri). Canadian Journal of Zoology, 65(10): 2421-2425. doi:10.1139/z87-365
  • Ndiaye, P., Forgue, J., Lamothe, V., Cauty, C., Tacon, P., Lafon, P., Davail, B., Fostier, A., Le Menn, F. & Nunez, J. (2006). Tilapia (Oreochromis niloticus) vitellogenins: development of homologous and heterologous ELISAs and analysis of vitellogenin pathway through the ovarian follicle. Journal of Experimental Zoology Part A: Comparative Experimental Biology, 305A(7), 576-593. ISSN: 0022-104X.
  • Roubal, W.T., Lomax, D.P., Willis, M.L. & Johnson, L.L. (1997). Purification and partial characterization of English sole (Pleuronectes vetulus) vitellogenin. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 118(3): 613-622. doi:10.1016/S0305-0491(97)00241-1
  • Routledge, E.J., Sheahan, D., Desbrow, C., Brighty, G.C., Waldock, M. & Sumpter, J.P. (1998). Identification of estrogenic chemicals in STW effluent 2: in vivo responses in trout and roach. Envıronmental Science and Technology, 32: 1559–1565. doi:10.1021/es970796a
  • Schiffmann, D. & De Boni, U. (1991). Dislocation of chromatin elements in prophase induced diethylstilbestrol: a novel mechanism by which micronuclei can arise. Mutation Research, 246(1): 113–122. doi:10.1016/0027-5107(91)90113-3
  • Shore, L., Gurevitz, M. & Shemesh, M. (1993). Estrogen as environmental pollutant. Bulletin of Environmental Contamination and Toxicology, 51: 361–366. doi:10.1007/BF00201753
  • Stumpf, M., Ternes, T.A., Ilaber, K. & Baumann, W. (1996). Determination of natural and synthetic estrogens in sewage plants and river water. Vom Wasser, 87: 251–261.
  • Sumpter, J.P. & Jobling, S. (1995). Vitellogenesis as biomarker for estrogenic contamination of the aquatic environment. Environmental Health Perspective, 103(7): 173–178. doi:10.2307/3432529
  • Ternes, T.A., Stumph, M., Mueller, J., Haberer, K., Wilken, R.D. & Servos, M. (1999). Behavior and occurrence of estrogens in municipal sewage treatment plants. I. Investigations in Germany, Canada and Brazil. Science of The Total Environment, 225: 81–90.
  • doi:10.1016/S0048-9697(98)00334-9
  • Tyler, C.R. & Sumpter, J. P. (1996). Oocyte growth and development in teleosts. Reviews in Fish Biology and Fisheries, 6: 287-318. doi:10.1007/BF00122584
  • Tyler, C.R. & Sumpter, J. P. (1990). The development of a radioimmunoassay for carp (Cyprinus carpio) vitellogenin. Fish Physiology and Biochemistry, 8(2): 129-140. doi:10.1007/BF00004440
  • Tyler, C.R., van der Eerden, B., Jobling, S., Panter, G. & Sumpter, J.P. (1996). Measurement of vitellogenin, a biomarker for exposure to oestrogenic chemicals, in a wide variety of cyprinid fish. Journal of Comparative Physiology B, 166: 418–426. doi:10.1007/BF02337886
  • Van den Belt, K., Verheyen, R. & Witters, H. (2003). Comparison of vitellogenin responses in zebrafish and rainbow trout following exposure to environmental estrogens. Ecotoxicology and Environmental Safety, 56: 271-281. doi:10.1016/S0147-6513(03)00004-6
  • Van den Belt, K., Verheyen, R. & Witters, H. (2001). Reproductive effects of ethynylestradiol and 4t-octylphenol on the zebrafish. Archives of Environmental Contamination and Toxicology, 41: 458–467. doi:10.1007/s002440010272
  • Yamanaka, Y., Mizuno,T., Sasai, Y., Kishi, M., Takeda, H., Kim C.H., Hibi, M. & Hirano, T. (1998). A novel homeobox gene, dharma, can induce the organizer in a non-cell-autonomous manner. Genes and Development, 12(15), 2345–2353. doi:10.1101/gad.12.15.2345
  • Yamaguchi, A., Ishibashi, H., Kohra, S., Arizono, K., Kato, K., Nakahama, T., Kanno, Y., Inouye, Y. & Tominaga, N. (2009). Expression analysis of estrogen-responsive genes vitellogenin 1 and 2 in liver of male medaka (Oryzias latipes) exposed to selective ligands of estrogen receptor subtypes. Journal of Health Science, 55(6): 930-938. doi:10.1248/jhs.55.930
There are 38 citations in total.

Details

Journal Section Articles
Authors

Göktuğ Gül

Aysel Çağlan Karasu Benli

Figen Erkoç

Publication Date May 15, 2016
Submission Date May 15, 2016
Published in Issue Year 2016Volume: 33 Issue: 2

Cite

APA Gül, G., Karasu Benli, A. Ç., & Erkoç, F. (2016). Mogan Gölü’ndeki bazı balık türlerinde vitellogenin proteininin elektroforetik karşılaştırılması. Ege Journal of Fisheries and Aquatic Sciences, 33(2), 151-156. https://doi.org/10.12714/egejfas.2016.33.2.09

Cited By

Mogan Gölü balık faunası
Balıkesir Üniversitesi Fen Bilimleri Enstitüsü Dergisi
https://doi.org/10.25092/baunfbed.321040