Haematological and Genotoxicological Research on Pelophylax ridibundus and Bufotes variabilis Living Around the Çan (Çanakkale, Turkey)

Year 2020, Volume: 4 Issue: 2, 105 - 111, 16.09.2020

Ceren Nur Özgül Didem Kurtul Çiğdem Gül

https://doi.org/10.26650/tjbc.20200011

Abstract

Haematologic parameters play an important role in the determination of the general health status as well as the physiology of amphibian species and the effects of various stress and poor conditions on some species. With this study, changes in the hematological parameters (red blood cell count, white blood cell count, hemoglobin concentration, hematocrit value, mean cell volume, mean cell hemoglobin and mean cell hemoglobin concentration) on an aquatic (Pelophylax ridibundus) and a terrestrial (Bufotes variabilis) amphibian living around the Çan (Çanakkale, Turkey) were presented. Also, to reveal the DNA damage on these two amphibian species, various nuclear abnormalities like micronucleus, binucleated nucleus, lobbed nucleus, blebbed, and notched nucleus were analyzed.
As a result of this study, the aquatic amphibian species when compared with the terrestrial amphibian species, significant differences in hemoglobin concentration (U= 22.5; W=88.5; Z= -3.446; p= 0.001), and heterophil count (U=34.00; W=205.00; Z= -3.79; p=0.00) were determined. The hemoglobin concentration of the terrestrial species was found higher while the heterophil count of the aquatic species was higher. Total nuclear abnormalities were found higher on the B. variabilis species but there was not a significant statistical difference. The micronucleus and other nuclear abnormalities of these two amphibian species when compared, the frequency of the notched nucleus was found statistically different. The frequency of the notched nucleus was higher in the B. variabilis species.

Keywords

Amphibia, hematology, micronucleus, Pelophylax ridibundus, Bufotes variabilis

Supporting Institution

This paper was supported by TÜBİTAK with 2209-A University Students Research Projects Support Program.

Project Number

2209-A

References

• Al-Sabti, K., & Metcalfe, C. D. (1995). Fish micronuclei for assessing genotoxicity in water. Mutation Research/Genetic Toxicology, 343(2-3), 121–135.
• Arserim, S. K., & Mermer, A. (2008). Hematology of the Uludag frog, Rana macrocnemis (Boulenger, 1885) in Uludag National Park (Bursa, Turkey). Ege University Journal of Fisheries Aquatic Sciences, 25, 39–46.
• Arvy, L. (1947). Le dimorphisme sexual sanguine chez Rana temporaria L. et Bufo vulgaris. Comptes Rendus Biologies, 141, 457–459.
• Atatür, M. K., Arıkan, H., & Çevik, I. E. (1999). Erythrocyte sizes of some Anurans from Turkey. Turkish Journal of Zoology, 23, 111–114.
• Banerjee, V. (1988). Erythrocytes related blood parameters in Bufo melanostictus with reference to sex and body weight. Environmental Ecology, Kalyani 6, 802–806.
• Başoğlu, M., & Öktem, N. (1984). Zoofizyoloji Praktikumu. Ege Üniversitesi Fen Fakültesi Kitaplar Serisi No: 41. 1–86.
• Bolognesi, C., Perrone, E., Roggieri, P., Pampanin, D. M., & Sciutto, A. (2006). Assessment of micronuclei induction in peripheral erythrocytes of fish exposed to xenobiotics under controlled conditions. Aquatic Toxicology, 78, S93–S98.
• Cabagna, M. C. Lajmanovich, R. C. Stringhini, G. Sanchez-Hermandez, J. C., & Peltzer, P. M. (2005). Hematological parameters of health status in the common toad Bufo arenarum in agroecosystems of Santa Fe province, Argentina. Applied Herpetology, 2, 373–380.
• Cunningham, W. P., Cunningham, M. A., & Saigo, B. W. (2001). Environmental science: A global concern (p. 452). New York: McGraw-Hill.
• Çördük, N. Tosunoğlu, M. Hacıoğlu Doğru, N., & Gül, Ç. (2018). Monitoring of micronuclei and nuclear abnormalities in Pelophylax ridibundus erythrocytes from the Biga Stream (Çanakkale, Turkey). Fresnius Environmental Bulettin, 27(1), 147–153.
• Dar, S. A., Yousuf, A. R., & Balkhi, M. (2016). An Introduction about Genotoxicology Methods as Tools for Monitoring Aquatic Ecosystem: Present Status and Future Perspectives. Fisheriers Aquatic Journal, 7, 1.
• Ergene, S., Çavaş, T., Celik, A., Köleli, N., Kaya, F., & Karahan, A. (2007). Monitoring of nuclear abnormalities in peripheral erythrocytes of three fish species from the
• Goksu Delta (Turkey): genotoxic damage in relation to water pollution. Ecotoxicology, 16(4), 385–391.
• Fenech, M. (2000). The in vitro micronucleus technique. Mutation Research, 455, 81–95.
• Goniakowska, L. (1973). Metabolism, resistance to hypotonic solutions, and ultrastructure of erythrocytes of five amphibian species. Acta Biologica Cracoviensia Series Zoologica, 13, 225–236.
• Guilherme, S., Válegab, M., Pereirab, M. E. Santosa, M.A., & Pachecoa, M. (2008). Erythrocytic nuclear abnormalities in wild and caged fish (Liza aurata) along an environmental mercury contamination gradient. Ecotoxicology and Environmental Safety, 70(3), 411–421.
• Gül, Ç., Tosunoğlu, M., Erdoğan, D., & Özdamar, D. (2011). Changes in the blood composition of some anurans. Acta Herpetologica, 6(2), 137–147.
• Gürkan, M., Hayretdağ, S., Yakın, B.Y., & Tok, C.V. (2012). A preliminary study on micronucleus analysis and nuclear anomalies in Pelophylax ridibundus (Pallas, 1771) (Amphibia: Anura) specimens collected around Vize (Kirklareli) and Ida Mountains (Çanakkale, Turkey). Ege University Journal of Fisheries Aquatic Science, 29(3), 133–136.
• Heddle J. A., & Countryman R. I. (1976). The production of micronuclei from chromosome aberration in irradiated cultures of human lymphocytes. Mutation Research, 41, 321–332.
• Hutchison, H. V., & Szarski, H. (1965). Number of erythrocytes in some Amphibians and Reptiles. Copeia, 3, 373–375.
• Josende, M. E., Tozetti, A. M., Alalan, M. T., Filho, V. M., Ximenez, S., Junior, F. M. R., & Martins, S. E. (2015). Genotoxic evaluation in two amphibian species from Brazilian subtropical wetlands. Ecological Indicators, 49, 83–87.
• Lajmanovich, R. C., Cabagna-Zenklusen, M. C., Attademo, A. M., Junges, C. M., Peltzer, P. M., Bassó, A., & Lorenzatti, E. (2014). Induction of micronuclei and nuclear abnormalities in tadpoles of the common toad (Rhinella arenarum) treated with the herbicides Liberty® and glufosinate-ammonium. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 769, 7–12.
• Marques, S. M., Antunes, S. C., Pissarra, H., Pereira, M. L., Gonçalves, F., & Pereira R. (2009) Histopathological changes and erythrocytic nuclear abnormalities in Iberian green frogs (Rana perezi Seoane) from a uranium mine pond. Aquatic Toxicology, 91, 187–195.
• Napierska, D., Barŝienė, J., Mulkiewicz, E., Podolska, M., & Rybakovas, A. (2009). Biomarker responses in flounder Platichthys flesus from the Polish coastal area of the Baltic Sea and applications in biomonitoring. Ecotoxicology, 18, 846–859.
• Özdamar, K. (2004). Paket Programlar ile İstatistiksel Veri Analizi (Çok Değişkenli Analizler), Eskişehir: Kaan Kitabevi.
• Rouf, M. A. (1969). Hematology of the leopard frog, Rana pipiens. Copeia, 682-687.
• Ruiz, G., Rosenmann, M. & Veloso, A. (1989). Altitudinal distribution and blood values in the toad, Bufo spinulosus Wiegmann. Comparative Biochemistry Physiology, 94A, 643–646.
• Schaumburg, L. G., Poletta, G. L., Siroski, P. A., & Mudry, M. D. (2012). Baseline values of Micronuclei and Comet Assay in the lizard Tupinambis merianae (Teiidae, Squamata). Ecotoxicology and Environmental Safety, 84, 99–103.
• Schermer, S. (1958). Die Blutmorphologie der Laboratoriumstiere. Barth.
• Sils, E. A. (2008). Specific of amphibian (genus Rana) peripheral blood leucogram under condition of anthropogenic load. The Problems of Herpetology. Saint-Petersburg, Russian Collection publishing, 369-374.Sinha, R.C. (1983). Hematological studies on the pre-wintering and wintering frog, Rana esculenta, Comp. Biochem. Physiol. 74, 311–314.
• Strunjak-Perovic, I. Lisicic, D., Coz-Rakovac, R., Jadan, M., Benkovic, V., & Tadic, Z. (2010). Evaluation of micronucleus and erythrocytic nuclear abnormalities in Balkan whip snake Hierophis gemonensis. Ecotoxicology, 19(8), 1460–1465.
• Szarski, H., & Czopek, G. (1966). Erythrocyte diameter in some amphibians and reptiles. Bulletin De L Academie Polonaise Des Sciences-Serie Des Sciences Biologiques, 14(6), 433.
• Şişman, T., Aşkın, H., Türkez, H., Özkan, H., İncekara, Ü., & Çolak. S. (2015). Determination of Nuclear Abnormalities in Peripheral Erythrocytes of the Frog Pelophylax ridibundus (Anura: Ranidae) sampled from Karasu River Basin (Turkey) for Pollution Impacts. LIMNOFISH-Journal of Limnology and Freshwater Fisheries Research, 1(2), 75–81.
• Tanyer, G. (1985). Hematoloji ve laboratuar, Ayyıldız Matbaası A.Ş. Ankara. 1–448.
• Udroiu, I. (2006). The micronucleus test in piscine erythrocytes. Aquatic Toxicology, 79(2), 201–204.
• Wojtaszek, J., & Adamowicz, A. (2003). Hematology of the fire bellied toad Bombina bombina L, Comparative Clinical Pathology, 12, 129–134.
• Wojtaszek, J., Baronowska, M., Glubiak, M., & Dzugaj, A. (1997). Circulating blood parameters of the water frog, Rana esculenta L. at pre-wintering stage. Zoologica Poloniae, (1-4), 117–126.
• Zhukova, T. I., & Kubantsev, B. S. (1979). Changes in the blood composition of salientians during hibernation. Sovyet Journal of Ecology, 9, 379–382.