Toxicological and physiological effects of ethephon on the model organism, Galleria mellonella L. 1758 (Lepidoptera: Pyralidae)

Year 2016, Volume: 40 Issue: 4, 0 - 0, 24.11.2016

Hülya Altuntaş Emine Duman Sümeyra Nur Şanal Demirci Ekrem Ergin

Abstract

Ethephon (ETF) has been used in agriculture as an ethylene releaser type of plant growth regulator. The aim of this work was to determine the ecotoxicological effects of ETF on the survival and the antioxidant metabolism of the insects using a model organism Galleria mellonella L. 1758. A toxicity test was performed to determine the lethal doses of ETF on larvae. According to probit assay, the LD₅₀ and LD₉₉ values for force fed larvae were 344 and 419 µg/5 µl, respectively, 30 d after treatment. Analyses performed with 10 doses ≤LD₅₀ at 24 and 48 h upon feeding larvae revealed that the malondialdehyde level increased at 300 and 330 µg/5 µl doses, whereas glutathione-S-transferase activity increased only with a 360 µg/5µl dose of ETF at 24 h. However, an increase in glutathione-S-transferase activity was evident at all ETF doses at 48 h. An increase in glutathione peroxidases activity was determined at 250, 300 and 330 µg/5µl at 24 and 48 h. All ETF doses caused an important increase in catalase activity at 24 h but remained unchanged at 48 h. Superoxide dismutase activity also elevated at doses >250 µg/5µl at 24 h when compared to the control. Same changes in superoxide dismutase activity were also observed at all doses of ETF except for 360 µg/5µl at 48 h. These results showed that ETF induced oxidative stress resulted in toxic effects that affected on the survival of model organism G. mellonella.

Keywords

Antioxidant enzymes, ethephon, Galleria mellonella, malondialdehyde, toxicology

Etefonun model organizma Galleria mellonella L. 1758 (Lepidoptera: Pyralidae) üzerine toksikolojik ve fizyolojik etkileri

Abstract

Etilen salınımına neden olan Etefon (ETF), bir bitki büyüme düzenleyicisi olarak tarımda kullanılmaktadır. Bu çalışmada bir model organizma olan Galleria mellonella L. 1758 (Lepidoptera: Pyralidae) türü kullanılarak, ETF’nin böceklerin antioksidan metabolizması ve canlılığı üzerindeki ekotoksikolojik etkilerinin araştırılması amaçlanmıştır. Larval dönemde ETF’nin letal dozunun belirlemesi amacıyla toksitite testi yapılmıştır. Larvalara zorla besleme (ağızdan besleme) yöntemi ile uygulanan ETF dozlarına göre, 30 günlük süreç içinde belirlenen LD₅₀ ve LD₉₉ değerleri sırasıyla 344 ve 419 µg/5µl olarak belirlenmiştir. LD₅₀ ve daha düşük ETF dozlarıyla yapılan toksikolojik analizlerde ise iki zaman dilimi (24. ve 48. saat) tercih edilmiştir. 24. saatte, larval hemolenfteki malondialdehit seviyesi, 300 ve 330 µg/5µl ETF dozlarında artarken, glutatyon-S-transferaz aktivitesi sadece 360 µg/5µ’lik dozda yükselmiştir. Ancak 48. saatte kontrol ve tüm dozlarda glutatyon-S-transferaz aktivitesi yükselmiştir. Glutatyon peroksidaz aktivitesi ise hem 24. hem de 48. saatte, 330 ve 360 µg/5µl ETF dozlarında artmıştır. Tüm ETF dozları 24. saatte katalaz aktivitesinde artışa neden olurken, bu artış 48. saatte de aynı kalmıştır. Süperoksit dismutaz aktivitesi ise 24. saatte, 250 µg/5µl ve daha yüksek dozlarda yükselmiştir. 48. saatteki süperoksit dismutaz aktivitesinde de benzer değişimler meydana gelirken, 360 µg/5µl dozunda azalma belirlenmiştir. Bu sonuçlar, ETF’nin oksidatif stresi teşvik etmesi sonucunda model organizma G. mellonella’nın canlılığı üzerinde toksik etkisi olduğunu göstermiştir.

Keywords

Antioksidan enzimler, etefon, Galleria mellonella, malondialdehit, toksikoloji

References

• Abd El Raouf, A. & S. M. Girgis, 2011. Mutagenic, teratogenic and biochemical effects of ethephon on pregnant mice and their fetuses. Global Veterinaria, 6 (3): 251-257.
• Ahmad, S. & R. S. Pardini, 1990. Mechanisms for regulating oxygen toxicity in phytophagous insects. Free Radical Biology and Medicine, 8: 401-413.
• Ahmad, S., 1995. Oxidative stress from environmental pollutants. Archives of Insect Biochemistry and Physiology. 29: 135-157.
• Ahmad, S., C. A. Pritsos, S. M. Bowen, C. R. Heisler, G. J. Blomquist & R. S. Pardini, 2005. Subcellular distrubition and activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase in the southern armyworm, Spodoptera eridania. Archives of Insect Biochemistry and Physiology, 7: 173-18.
• Altuntaş, H., A. Y. Kılıç, F. Uçkan & E. Ergin, 2012. Effects of gibberellic acid on hemocytes of Galleria mellonella L. (Lepidoptera: Pyralidae). Environmental Entomology, 41 (3): 688-696.
• Altuntaş, H., F. Uçkan, A. Y. Kılıç, & E. Ergin, 2014. Effects of gibberellic acid on hemolymph free amino acids of Galleria mellonella (Lepidoptera: Pyralidae) and endoparasitoid Pimpla turionellae (Hymenoptera: Ichneumonidae). Annals of the Entomology Society of America, 107 (5): 1000-1009.
• Altuntaş, H., 2015a. Determination of gibberellic acid (GA3)-induced oxidative stress in a model organism Galleria mellonella L. (Lepidoptera: Pyralidae). Environmental Entomology (Physiology), 44 (1): 100-105, DOI: 10.1093/ee/nvu020.
• Altuntaş, H., 2015b. Effects of ethephon on the hemolymph metabolites of the greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae). Acta Physica Polonica A, 128: 182-183, DOI: 10. 12693/AphysPolA.128.B-182.
• Al-Twaty, N. H. A., 2006. Mutagenic effects of ethephon on albino mice. Journal of Biological Sciences, 6 (6): 1041-1046.
• Anant, J. D. & B. G. Avinash, 2012. Modulation in serum biochemicals in European rabbit, Oryctolagus cuniculus (Linn.) exposed to ethephon. European Journal of Experimental Biology, 2 (3): 794-799.
• Aslantürk, A., S. Kalender, M. Uzunhisarcıklı & Y. Kalender, 2011. Effects of methidathion on antioxidant enzyme activities and malondialdehyde level in midgut tissues of Lymantria dispar (Lepidoptera) larvae. Journal of the Entomological Research Society, 13 (3): 27-38.
• Bhadoria, P., M. Mahindra, V. Bahrioke & A. S. Bhadoria, 2015. Effect of ethephon on the liver in albino rats: a histomorphometric study. Biomedical Journal, 38: 421-427.
• Bui, Q. Q., 2007. Ethephon and jackfruit, Palo Alto, California. 231pp.
• Bradford, M. M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254.
• Bronskill, J. F., 1961. A cage to simplify the rearing of thegreater wax moth, Galleria mellonella (Pyralidae). Journal of Lepidopterists’ Society, 15: 102-104.
• Büyükgüzel, E., P. Hyrsl & K. Büyükgüzel, 2010. Eicosanoids mediate hemolymph oxidative and antioxidative response in larvae of Galleria mellonella L. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 156: 176-83.
• Büyükgüzel, E., K. Büyükgüzel, M. Snela, M. Erdem, K. Radtke, K. Ziemnicki, & Z. Adamski, 2013. Effect of boric acid on antioxidant enzyme activity, lipid peroxidation and ultrastructure of midgut and fat body of Galleria mellonella. Cell Biology and Toxicology, 29: 117-129.
• Chance, B. & A. C. Maehly, 1955. Assay of catalase and peroxidases. Methods Enzymology, 2: 764-775.
• Cook, S. M. & J. D. McArthur, 2013. Developing Galleria mellonella as amodel host for human pathogens. Virulence, 4 (5): 350–353.
• Dere, B., H. Altuntaş, & Z. U. Nurullahoğlu, 2015. Insecticidal and oxidative effects of azadirachtin on the model organism Galleria mellonella l. (Lepidoptera: Pyralidae). Archives of Insect Biochemistry and Physiology, DOI:10.1002/arch.21231.
• Emre, İ., T. Kayış, M. Coşkun, O. Dursun & H. Y. Cogun, 2013. Changes in antioxidative enzyme activity, glycogen, lipid, protein, and malondialdehyde content in cadmium-treated Galleria mellonella larvae. Annals of the Entomology Society of America, 106 (3): 371-377.
• Erdem, M. & E. Büyükgüzel, 2015. The Effects of xanthotoxin on the biology and biochemistry of Galleria mellonella L. (Lepidoptera: Pyralidae). Archives of Insect Biochemistry and Physiology, DOI: 10.1002/arch.21236.
• Fahmy, N. M., 2012. Impact of two insect growth regulators on the enhancement of oxidative stress and antioxidant efficiency of the cotton leaf worm, Spodoptera littoralis (Biosd.). Egyptian Academic Journal of Biological Sciences, 5 (1): 137-149.
• Felton, G. W. & C. B. Summers, 1995. Antioxidant systems in insects. Archives of Insect Biochemistry and Physiology, 29: 187-97.
• Gupta, G., S. R. Yadav & A. K. Bhattacharya, 2009. Influence of synthetic plant growth substances on the survivorship and developmental parameters of Spilarctia obliqua Walker (Lepidoptera: Arctiidae). Journal of Pesticide Science, 82: 41-46.
• Haux, J. E., G. B. Quistad & J. E. Casida, 2000. Phosphobutyrylcholinesterase: Phosphorylation of the Esteratic Site of Butyrylcholinesterase by Ethephon [(2- Chloroethyl) phosphonic acid] Dianion. Chemical Research in Toxicology, 13: 646-651.
• Haux, J. E., O. Lockridge & J. E. Casida, 2002. Specificity of ethephon as a butyrylcholinesterase inhibitor and phosphorylating agent. Chemical Research in Toxicology, 15: 1527-1533.
• Hemming, J. D. C. & R. Lindroth, 2000. Effects of phenolic glycosides and protein on gypsy moth (Lepidoptera : Lymantriidae) and forest tent caterpillar (Lepidoptera : Lasiocampidae) performance and detoxification activities. Environmental Entomology, 2: 1108-1115.
• Henneberry, T. J., T. Meng, W. D. Hutchison, L. A. Bariola & B. Deeter, 1988. Effects of ethephon on boll weevil (Coleoptera: Curculionidae) population development, cotton fruiting, and boll opening. Journal of Economic Entomology, 81 (2): 628-633.
• Hussain, I., A. Saeed, A. Muhammad & A. Rashid, 2015. Ethephon application at kimri stage accelerates the fruit maturation period and improves phytonutrients status (Hillawi and Khadrawi (c.v.)) of date palm fruit. Pakistan Journal of Agricultural Sciences, 52 (2): 415−423.
• Hyrsl, P., E. Büyükgüzel & K. Büyükgüzel, 2007. The effects of boric acid-induced oxidative stress on antioxidant enzymes and survivorship in Galleria mellonella. Archives of Insect Biochemistry and Physiology, 66: 23-31.
• İçen, E., F. Armutçu, K. Büyükgüzel & A. Gürel, 2005. Biochemical stress indicators of greater wax moth exposure to organophosphorus insecticides. Journal of Economic Entomology, 98: 358-366.
• Kaur, R. & P. J. Rup, 2003. Influence of four plant growth regulators on development of the melon fruit fly, Bactrocera cucurbitae (Coquillett). Insect Science and Its Application, 23: 121-125.
• Krishnan, N. & D. Kodrik, 2006. Antioxidant enzymes in Spodoptera littoralis (Boisduval): Are they enhanced to protect gut tissues during oxidative stress? Journal of Insect Physiology, 52: 11-20.
• Maguire, R., O. Duggan, & K. Kavanagh, 2016. Evaluation of Galleria mellonella larvae as an in vivo model for assessing the relative toxicity of food preservative agents. Cell Biology and Toxicology, DOI: 10.1007/s10565-016-9329-x.
• Oruc, E., 2011. Effects of diazinon on antioxidant defense system and lipid peroxidation in the liver of Cyprinus carpio (L.). Environmental Toxicology, 26: 571-578.
• Peric´-Mataruga, V., D. Blagojevic, M.B. Spasic, J. Ivanovic & M. Jankovic-Hladni, 1997. Effect of the host plant on the antioxidative defence in the midgut of Lymantria dispar L. caterpillars of different population origins. Journal of Insect Physiology, 43: 101-106.
• Riberio, S., J. P. Sousa, J. A. A. Nogueira & A. M. V. M. Soares, 2001. Effect of endosulfan and parathion on energy reserves and physiological parameters of the terrestrial isopod Porcellio dilatatus. Ecotoxicology and Environmental Safety, 49: 131-138.
• Sancho, E., M. D. Ferrando, C. Fernandez & E. Andreu, 1998. Liver energy metabolism of Anguilla anguilla after exposure to fenitrothion. Ecotoxicology and Environmental Safety, 41: 68-175.
• SPSS, Inc., SPSS 18.0 Statistics. SPSS, Chicago, IL (2010).
• Stotz, H. U., B. R. Pittendrigh, J. Kroymann, K. Weniger, J. Fritsche, A. Bauke & T. Mitchell-Olds, 2000. Induced plant defense responses against chewing insects ethylene signaling reduces resistance of arabidopsis against egyptian cotton worm but not diamondback moth. Plant Physiology, 124: 1007-1017.
• Tuluce, Y. & I. Celik, 2006. Influence of subacute and subchronic treatment of abcisic acid and gibberellic acid on serum marker enzymes and erythrocyte and tissue antioxidant defense systems and lipid peroxidation in rats. Pesticide Biochemistry and Physiology, 86: 85-92.
• Uçkan, F., A. Tüven, A. Er & E. Ergin, 2008. Effects of gibberellic acid on biological parameters of the larval endoparasitoid Apanteles galleriae (Hymenoptera: Braconidae). Annals of the Entomology Society of America, 101: 593-597.
• Uçkan, F., İ. Haftacı & E. Ergin, 2011a. Effects of indol-3-acetic acid on biological parameters of the larval endoparasitoid Apanteles galleriae (Hymenoptera: Braconidae). Annals of the Entomology Society of America, 104 (1): 77-82.
• Uçkan, F., Z. Öztürk, H. Altuntaş & E. Ergin, 2011b. Effects of gibberellic acid (GA3) on biological parameters and hemolymph metabolites of the pupal endoparasitoid Pimpla turionellae (Hymenoptera: Ichneumonidae) and its host Galleria mellonella (Lepidoptera: Pyralidae). Journal of the Entomological Research Society, 13: 1-14.
• Uçkan, F., H. K. Soydabaş & R. Özbek, 2014. Effect of indol-3 acetic acid on the biochemical parameters of Achoria grisella hemolymph and Apanteles galleriae larva. Pakistan Journal of Biotechnology, 11 (2): 163-171.
• Uçkan, F., R. Özbek & E. Ergin, 2015. Effects of indol-3-acetic acid on the biology of Galleria mellonella and its endoparasitoid Pimpla turionellae. Belgian Journal of Zoology, 145 (1): 49-58.
• Wang, K. S., C. Y. Lu & S. H. Chang, 2011. Evaluation of acute toxicity and teratogenic effects of plant growth regulators by Daphnia magna embryo assay. Journal of Hazardous Materials, 190: 520-528, DOI:10.1016/j.jhazmat.2011.03.068.
• Yu, S. J., 2004. Induction of detoxification enzymes by triazine herbicides in the fall armyworm, Spodoptera frugiperda. Pesticide Biochemistry and Physiology, 80: 113-122.
• Zhang, L., S. Li, X. Liu, C. Song & X. Liu, 2012. Effects of ethephon on physicochemical and quality properties of kiwifruit during ripening. Postharvest Biology and Technology, 65: 69−75.