Hüseyin ARIKAN, Bayram GÖÇMEN, Naşit İĞCİ, Bahadır AKMAN

Age-dependent variations in the venom proteins of Vipera kaznakovi Nikolsky, 1909 and Vipera ammodytes (Linnaeus, 1758) (Ophidia: Viperidae)

Atatür (1979) as previously described in detail (Arıkan et al., 2006). Briefly, venom samples were loaded onto the polyacrylamide gels and separated using a Canalco model 1200 electrophoresis apparatus. Gels were stained with 0.5% Amido black to visualize the protein bands and densitometric curves were obtained using a Gelman ACD-15 model 39430 densitometer at 500 nm.The authors received ethical permission for venom sampling (Ege University Animal Experiments Ethics Committee, 2010-43) and special permission for field studies (from Republic of Turkey Ministry of Forestry and Water Affairs).3. ResultsThe venom extracts of Vipera kaznakovi and Vipera ammodytes specimens had a viscosity higher than that of water. Although venom secretions of 2 specimens of V. kaznakovi (16.5 and 30 cm total lengths) were colorless, they were light yellow in the 55-cm-long specimen. In contrast, the venom extracts of 3 specimens of V. ammodytes were light yellow.The gel photographs of the venom protein samples of V. kaznakovi specimens are presented in Figure 1. According to our results, the venom proteins of V. kaznakovi were separated into 12 fractions or fraction groups in the 2 shorter individuals (16.5 and 30 cm), while there were 13 fractions or fraction groups in the gel of the 55-cm-long individual (the longest one). Our comparative results of electrophoretic density curves (protein fractions) revealed significant quantitative differences especially between the 2 shorter individuals, while similar electrophoretic mobility patterns and fraction numbers were observed (Figures 1–4).

Anahtar Kelimeler:

Vipera ammodytes, polyacrylamide disc gel electrophoresis, densitometry

Age-dependent variations in the venom proteins of Vipera kaznakovi Nikolsky, 1909 and Vipera ammodytes (Linnaeus, 1758) (Ophidia: Viperidae)

Polyacrylamide disc gel electrophoresis and densitometry analysis were used to analyze venom extracts of Vipera kaznakovi (16.5, 30, and 55 cm) and Vipera ammodytes (28.5, 36.7, and 51.5 cm) specimens of 3 different lengths. V. kaznakovi specimens were collected from Hopa (Artvin Province); 2 V. ammodytes specimens were from Ağva (İstanbul Province) and the longest one was from Yuvacık (Kocaeli Province). The electropherograms of the venom protein samples showed age-dependent qualitative and quantitative variations.

Keywords:

Vipera ammodytes polyacrylamide disc gel electrophoresis, densitometry, Vipera kaznakovi,

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Arıkan H, Kumlutaş Y, Türkozan O, Baran İ (2003). Electrophoretic patterns of some viper venoms from Turkey. Turk J Zool 27: 239‒242.
Arıkan H, Göçmen B, Mermer A, Bahar H (2005). An electrophoretic comparison of the venoms of a colubrid and various viperid snakes from Turkey and Cyprus, with some taxonomic and phylogenetic implications. Zootaxa 1038: 1‒10.
Arıkan H, Keskin NA, Çevik İE, Ilgaz Ç (2006). Age-dependent variations in the venom proteins of Vipera xanthina (Gray, 1849) (Ophidia: Viperidae). Turkiye Parazitol Derg 30: 163‒165.
Arıkan H, Göçmen B, Kumlutaş Y, Alpagut-Keskin N, Ilgaz Ç, Yıldız MZ (2008). Electrophoretic characterisation of the venom samples obtained from various Anatolian snakes (Serpentes: Colubridae, Viperidae, Elapidae). North-Western Journal of Zoology 4: 16–28.
Bonilla CA, Horner NV (1969). Comparative electrophoresis of Crotalus and Agkistrodon venoms from North American snakes. Toxicon 7: 327‒329.
Bonilla CA, Faith MR, Minton SA (1973). L-amino acid oxidase, phosphodiesterase, total protein and other properties of juvenile timber rattlesnake (C. h. horridus) venom at different stages of growth. Toxicon 11: 301‒303.
Budak A, Göçmen B (2008). Herpetoloji. 2nd edition. İzmir, Turkey: Ege Üniversitesi Yayınları Fen Fakültesi Yayın No: 194 (in Turkish).
Chippaux J-P, Williams V, White J (1991). Snake venom variability: Methods of study, results and interpretation. Toxicon 29: 1279‒1303.
Chippaux J-P (2006). Snake Venoms and Envenomations. Malabar, FL, USA: Krieger Publishing Company.
Davis BJ (1964). Disc Electrophoresis-II, Method and Application to Human Serum Proteins. Ann N Y Acad Sci 121: 404‒427.
Dimitrov GD, Kankonkar RC (1968). Fractionation of Vipera russelli venom by gel filtration‒II, comparative study of yellow and white venoms of Vipera russelli with special reference to the local necrotizing and local actions. Toxicon 5: 283‒288.
Fiero MK, Seifert MW, Weaver TJ, Bonilla CA (1972). Comparative study of juvenile and adult prairie rattlesnake (Crotalus viridis viridis) venoms. Toxicon 10: 81‒82.
Furtado MFD, Maruyama M, Kamiguti AS, Antonio LC (1991). Comparative study of nine Bothrops snake venoms from adult female snakes and their offspring. Toxicon 29: 219‒226.
Glenn JL, Straight RC (1985). Venom properties of the rattlesnakes (Crotalus) inhabiting the Baja California region of Mexico. Toxicon 23: 769‒775.
Göçmen B, Arıkan H, Mermer A, Langerwerf B, Bahar H (2006). Morphological, hemipenial and venom electrophoresis comparisons of the Levantine viper, Macrovipera lebetina (Linnaeus, 1758), from Cyprus and southern Anatolia. Turk J Zool 30: 225–234.
Guércio RAP, Shevchenko A, Shevchenko A, López-Lozano JL, Paba J, Sousa MV, Ricart CAO (2006). Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox. Proteome Science 4: 11.
Igci N, Ozel Demiralp D (2012). A preliminary investigation into the venom proteome of Macrovipera lebetina obtusa (Dwigubsky, 1832) from Southeastern Anatolia by MALDI-TOF mass spectrometry and comparison of venom protein profiles with Macrovipera lebetina lebetina (Linnaeus, 1758) from Cyprus by 2D-PAGE. Arch Toxicol 86: 441‒451.
Jiménez-Porras JM (1964a). Intraspecific variations in composition of venom of the jumping viper, Bothrops nummifera. Toxicon 2: 187‒195.
Jiménez-Porras JM (1964b). Venom proteins of the Fer-de-Lance, Bothrops atrox, from Costa Rica. Toxicon 2: 155‒166.
Klauber LM (1956). Rattlesnakes: Their habits, life histories and influence on mankind. Berkeley, USA: University of California Press.
Mackessy SP (1988). Venom ontogeny in the Pacific rattlesnakes Crotalus viridis helleri and C. v. oreganus. Copeia 1988: 92‒101.
Mackessy SP, Williams K, Ashton, KG (2003). Ontogenetic variation in venom composition and diet of Crotalus oreganus concolor: a case of venom paedomorphosis? Copeia 2003: 769‒782.
Mackessy SP, Sixberry NP, Heyborne WH, Fritts T (2006). Venom of the brown treesnake, Boiga irregularis: ontogenetic shifts and taxa-specific toxicity. Toxicon 47: 537–548.
Mackessy SP (2010). The Field of Reptile Toxinology: Snakes, Lizards, and Their Venoms. In: Mackessy SP, editor. Handbook of Venoms and Toxins of Reptiles. Boca Raton, FL, USA: CRC Press Taylor & Francis Group, pp. 3‒23.
Mallow D, Ludwig D, Nilson G (2003). True Vipers: Natural History and Toxinology of Old World Vipers. Malabar, FL, USA: Krieger Publishing Company.
Meier J, Freyvogel TA (1980). Comparative studies on venoms of the Fer-de-lance (Bothrops atrox), carpet viper (Echis carinatus) and spitting cobra (Naja nigricollis) snakes at different ages. Toxicon 18: 661‒662.
Meier J (1986). Individual and age-dependent variations in the venom of the Fer-de-lance (Bothrops atrox). Toxicon 24: 41‒46.
Minton SA (1967). Observations on toxicity and antigenic make up of venoms from juvenile snakes. In: Russell FE and Saunders PR, editors. Animal Toxins. Oxford, UK: Pergamon Press, pp. 211‒222.
Minton SA (1975). A note on the venom of an aged rattlesnake. Toxicon 13: 73‒74.
Nalbantsoy A, Baykan Erel Ş, Köksal Ç, Göçmen B, Yıldız MZ, Karabay Yavaşoğlu NÜ (2013). Viper venom induced inflammation with Montivipera xanthina (Gray, 1849) and the anti-snake venom activities of Artemisia absinthium L. in rat. Toxicon 65: 34‒40.
Özeti N, Atatür MK (1979). A preliminary survey of the serum proteins of a population of Mertensiella luschani finikensis Başoğlu and Atatür, 1975 from Finike in Southwestern Anatolia. Istanbul University Journal of Faculty of Science 44B: 23‒
Tan N-H, Fung S-Y (2010). Snake Venom L-Amino Acid Oxidases. In: Mackessy SP, editor. Handbook of Venoms and Toxins of Reptiles. Boca Raton, FL, USA: CRC Press Taylor & Francis Group, pp. 221‒235.
Tare TG, Sutar NK, Renapurkar DM (1986). A study of snake venom yield by different methods of venom extraction. AmphibiaReptilia 7: 187‒191.
Theakston RDG, Reid HA (1978). Changes in the biological properties of venom from Crotalus atrox with aging. Period Biol 80: 123‒133.
Tomovic L (2006). Systematics of the nose-horned viper (Vipera ammodytes, Linnaeus, 1758). Herpetological Journal 16: 191‒201.
Tu AT (1996). Overview of snake venom chemistry. In: Singh BR, Tu AT, editors. Natural Toxins II. New York, NY, USA: Plenum Press, pp. 37‒62.
Tun-Pe, Nu-Nu-Lwin, Aye-Aye-Myint, Kyi-May-Htwe, KhinAung-Cho (1995). Biochemical and biological properties of the venom from Russell’s viper (Daboia russelli siamensis) of varying ages. Toxicon 33: 817‒821.
Yalcin HT, Ozen MO, Gocmen B, Nalbantsoy A (2013). Effect of Ottoman viper (Montivipera xanthina (Gray, 1849)) venom on various cancer cells and on microorganisms. Cytotechnology DOI: 1007/s10616-013-9540-z (in press).