Toxicological effects of the entomopathogenic Purpureocillium lilacinus on the model organism, Galleria mellonella

Year 2017, Volume: 10 Issue: 1, 153 - 159, 15.04.2017

Hülya Altuntaş Emine Duman

Abstract

The toxicological effects of entomopathogenic fungus Purpureocillium lilacinum Thom Luangsa-ard, Houbraken, Hywel-Jones and Samson on the Galleria mellonella L. Lepidoptera: Pyralidae larvae which have great importance in biological, physiological and biochemical studies as a model insect were investigated. For this purpose, larvae were injected with median lethal concentration LC50 2.82 x 103 conidia/ml suspension. Hemolymph samples were collected after injection at 24, 48, 72 and 96 hours. Total protein analysis and specific enzyme activities of CAT and GST in the hemolymph were analyzed spectrophotometrically. Total protein amount in the hemolymph of the infected larvae decreased at all times when compared with control groups. Furthermore, the activity of GST in the hemolymph of the infected larvae increased at all times compared with untreated larvae, but the activity of CAT was similar with activity of untreated larvae. Consequently, increasing the activity of GST showed that entomopathogen organisms are effective on the antioxidant defense system of insects as a source of free radical. CAT activity in the absence of any change showed inactivation of CAT out of increased superoxide radicals based pathogenicity. Therefore, we suggest that the variance in the activity of CAT and GST in larvae may indicate a physiological adaptability to compensate for pathogen-induced stress

Keywords

Galleria mellonella, Purpureocillium lilacinus, entomopathogen fungus, catalase, glutathione S-transferase

Entomopatojenik Purpureocillium lilacinus’un model organizma, Galleria mellonella üzerindeki toksikolojik etkileri

Abstract

Biyolojik, fizyolojik ve biyokimyasal araştırmalarda model organizma olarak büyük öneme sahip olan Galleria mellonella L. Lepidoptera: Pyralidae larvaları üzerinde entomopatojenik fungus Purpureocillium lilacinum Thom Luangsa-ard, Houbraken, Hywel-Jones ve Samson’un toksikolojik etkileri araştırılmıştır. Bu amaçla ortalama letal konsantrasyon LC50 olan 2.82 x 103 spor/ml süspansiyonu kullanılarak son evre larvalara enjeksiyon yapıldı. Enjeksiyondan 24, 48, 72 ve 96 saat sonra larvalardan hemolenf alındı. Hemolenf örneklerinde toplam protein miktarı, katalaz CAT ve glutatyon S-transferaz GST aktiviteleri spektrofotometrik olarak analiz edildi. P. lilacinus ile enfekte olan larvaların toplam protein miktarı tüm saatlerde kontrol grupları ile karşılaştırıldığında azalış göstermektedir. Ayrıca fungal enfeksiyona bağlı olarak GST enzim aktivitesi zamanla artış gösterirken, CAT enzim aktivitesinde herhangi bir değişiklik olmadığı belirlenmiştir. Bu sonuçlar, larval GST aktivitesinde görülen artışın patojenite kaynaklı oksidatif stresin telafi edilebilmesi için fizyolojik bir uyum olabileceğine, CAT aktivitesinde hiçbir değişimin olmaması ise patojenite kaynaklı artan süperoksit radikallerinin enzimin inaktivasyonuna neden olabileceğine işaret etmektedir. Bu nedenle, CAT ve GST aktivitesinde görülen bu değişikliklerin nedeninin patojen kaynaklı strese karşı oluşan fizyolojik onarım mekanizmalarıyla ilişkili olduğunu düşünmekteyiz

Keywords

Galleria mellonella, Purpureocillium lilacinus, entomopatojen fungus, katalaz, glutatyaon-Stranferaz

References

• Altuntaş, H., Duman, E., Şanal-Demirci, S. N., Ergin, E. (2016). Toxicological and physiological effects of ethephon on the model organism, Galleria mellonella L. 1758 (Lepidoptera: Pyralidae). Türkiye Entomoloji Dergisi, 40(4), 413- 423.
• Boyland, E., Chasseaud, L.F. (1969). The Role of Glutathione and Glutathione S Transferase in Mercaptic Acid Biosynthesis. Advances in Enzymology, 32, 173-219.
• Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principal of protein-dye binding. Annual Review of Biochemistry, 72, 248-254.
• Bronskill, J. F. (1961). A cage to simplify the rearing of the greater wax moth, Galleria mellonella (Pyralidae). The Journal of the Lepidopterists Society, 15, 102-104.
• Chance, B., Maehly, A. C. (1955). Assay of catalase and peroxidases. Methods Enzymology, 2, 764-775.
• Cook, S. M., McArthur, J. D. (2013). Developing Galleria mellonella as a model host for human pathogens. Virulence, 4, 350-353.
• Deacon, J.W. (1983). Microbial Control of Pests and Diseases. New York, 31-41.
• Demirel, R., Ilhan, S., Asan, A., Kınacı, E., Oner, S. (2005). Microfungi in cultivated fields in Eskişehir provience (Turkey). Journal of Basic Microbiology, 45, 279-293.
• Dere, B., Altuntaş, H., Nurullahoğlu, Z. U. (2015). Insecticidal And Oxidatıve Effects Of Azadırachtın On The Model Organism Galleria mellonella L. (Lepidoptera: Pyralidae). Archives of Insect Biochemistry and Physiology, 0, 1-15.
• Emre, I., Kayis, T., Coskun, M., Dursun, O., Cogun, H. Y. (2013). Changes in antioxidative enyzme activity, glycogen, lipid, protein, and melondialdehyde content in cadmium-treated Galleria mellonella larvae. Annals of the Entomological Society of America, 106, 371-377.
• Ergin, E., Altuntaş, H., Uçkan, F. (2013). Effects of parasitization and envenomation by the endoparasitic Wasp Pimpla turionellae L. (Hymenoptera: Ichneumonidae) on hemolymph protein profile of its host Galleria mellonella L. (Lepidoptera: Pyralidae). Biological Diversity and Conservation, 6(1), 62-70.
• Felton, G. W., Duffey, S. S. (1991). Protective action of midgut catalase in lepidopteran larvae against oxidative plant defenses. Journal of Chemical Ecology, 17(9), 1715-1732.
• Felton, G. W., Summers, C. B. (1995). Antioxidant systems in insects. Archives of Insect Biochemistry and Physiology, 29, 187-197.
• Feyereisen, R. (1999). Insect P450 Enzymes. Annual Review of Entomology, 44, 507-533.
• Fuchs, B. B., O’Brien, B. E., El Khoury, J. B., Mylonakis, E. (2010). Methods for using Galleria mellonella as a model host to study fungal pathogenesis. Virulence, 1(6), 475-482.
• Fuchs, S. Y., Spiegelman, V. S., Belitsky, G. A. (1993). The Effect of the Cytochrome P-450 System Inducers on the Development of Drosophila melanogaster. Journal of Biochemical and Molecular Toxicology, 8, 83-88.
• Glupov, V. V., Slepneva, I. A., Serebrov V. V., Khvoschevskay, M. F., Martem’yanov, V. V., Dubovskiy, I. M., Khramtsov, V. V. (2003). Influence of the Fungal Inflection on the Production of Reactive Oxygen Metabolites and the Antioksidant State of Haemolymph of Galleria mellonella L. (Lepidoptera: Pyralidae) Larvae. Russian Entomological Journal, 12( 1), 103-108.
• Goryunova, T. E., Vainer, L. M., Sidorov, V. N., vd. (1991). Use of [3H]-Permethrin to Evaluate Esterase Activity in Caterpillars of Beet Webworm Pyrausta sticticalis L. Agrokhimiya, 2, 118-121.
• Greathouse, K. L., Samuels, M., DiMarco, N. M., Criswell, D. S. (2005). Effects of Increased Dietary Fat and Exercise on Skeletal Muscle Lipid Peroxidation and Antioxidant Capacity in Male Rats. European Journal of Nutrition, 44(7), 429-35.
• Grubor-Lajsic, G., Block, W., Telesmanic, M., Jovanovic, A., Stevanovic, D., Baca, F. (1997). Effect of Cold Acclimation on the Antioxidant Defense System of Two Larval Lepidoptera (Noctuidae). Archives of Insect Biochemistry and Physiology, 36, 1-10.
• Halkman, A. K. (2005). Gıda Mikrobiyolojisi Uygulamaları. Başak Matbaacılık ve Tanıtım Hizmetleri Ltd. Şti., Ankara.
• Halliwell, B., Gutteridge, J. M. C. (2007). Free Radicals in Biology and Medicine. Oxford University Press, New York, USA.
• Hughes, W. O., Thomsen, L., Eilenberg, J., Boomsma, J. J. (2004). Diversity of Entomopathogenic Fungi Near Leaf- Cutting Ant Nests in a Neotropical Forest, with Particular Reference to Metarhizium anisopliae var. Anisopliae. Journal of Invertebrate Pathology, 85, 46-53.
• İçen, E., Armutçu, F., Büyükgüzel, K., Gürel, A. (2005). Biochemical stress indicators of greater wax moth exposure to organophosphorus insecticides. Journal of Economical Entomology, 98(2), 358-366.
• Kol’chevskaya, E. N., Kol’chevkii, A. G. (1988). Analysis of Isozymes of Nonspecific Esterases in Cabbage Moth Infected with Vairimorpha antheraeae Microsporidia. Byull, VIZR, 71.
• Kono, Y., Fridovich, I. (1982). Superoxide Radical Inhibit Catalase. Journal of Biological Chemistry, 257, 5751-5754.
• Krishnan, N., Kodrik, D. (2006). Antioxidant enzymes in Spodoptera littoralis (Boisduval) are they enhanced to protect gut tissues during oxidative stress. Journal of Insect Physiology, 5, 11-20.
• Krishnan, N., Sehnal, F. (2006). Compartmentalization of oxidative stress and antioxidant defense in the larval gut of Spodoptera littoralis. Archives of Insect Biochemistry and Physiology, 63, 1-10.
• Lozinskaya, Y. L., Slepneva, I. A., Khramtsov, V. V., Glupov, V. V. (2004). Changes of the Antioxidant Status and System of Generation of Free Radicals in Hemolymph of Galleria mellonella Larvae at Microsporidiosis. Journal of Evolutionary Biochemistry and Physiology, 40(2), 119-125.
• Pedro, H. C., Candido, S. A. (1997). Entomopathogenic Fungi Associated with Natural Populations of the Moroccan Locust Dociostaurus maroccanus (Thunberg) (Orthoptera: Gomphocerinae). Technology, 7, 357-363.
• Pitt, J. I. (1979). The Genus Penicillium and its telemorphic states Eupenicillium and Talaromyces. Academic Pres INC, London.
• Samashekaraiah, B. V., Padmaja, K., Prasad, A. R. K. (1992). Lead-Induced Lipid Peroxidation and Antioxidant Defense Components of Developing Chick Embryos. Free Radical Biology and Medicine, 13, 107-114.
• Sanal S., Altuntaş H., Kılıç Y. (2011). Effects of Fungal Infection by The Entomopathogen Fungus Paecilomyces lilacinus on Hemolymph Protein Profile of the Wax Moth Galleria mellonella L. (Lepidoptera: Pyralidae) Larvae. Sixth International Symposium on Molecular Insect Science, Amsterdam, The Netherlands.
• Sanjaya, Y., Ocampo V. R., Caoili B. L. (2016). Pathogenicity of three entomopathogenic fungi, Metarhizium anisopliae, Beauveria bassiana, and Paecilomyces lilacinus, to Tetranychus kanzawai infesting papaya seedlings. Arthropods, 5(3),109-113.
• Serebrov, V. V., Gerber, O. N., Malyarchuk, A. A., Martemyanov, V. V., Alekseev, A. A., Glupov, V. V. (2006). Effect of Entomopathogenic Fungi on Detoxification Enzyme Activity in Greater Wax Moth Galleria mellonella L. (Lepidoptera, Pyralidae) and Role of Detoxification Enzymes in Development of Insect Resistance to Entomopathogenic Fungi. Biology Bulletin, 33(6) 581-586.
• Serebrov, V. V., Kiselev, A. A., Glupov, V. V. (2003). Study of Some Factors of Synergy between Entomopathogenic Fungi and Chemical Insecticides. Mikol Fitopatol, 37(1), 76-82.
• Sies, H. (1997). Oxidative stress: oxidants and antioxidants. Experimental Physiology, 82, 291-295.
• Terriere, L. C. (1984). Induction of Detoxication Enzymes in Insects. Annual Review of Entomology, 29, 71-88.
• Urso, M. L., Clarkson, P. M. (2003). Oxidative Stress, Exercise and Antioxidant Supplementation. Toxicology. 189(1- 2), 41-54.
• Wu, G., Jiang, S., Miyata, T. (2004). Effects of Synergists on Toxicity of Six Insecticides in Parasitoid Diaeretiella rapae (Hymenoptera: Aphidiidae). Journal of Economic Entomology, 97, 2057-2066.
• Xia, Y., Dean, P., Judge, A. J., Gillespie, J. P., Clarkson, J. M., Charnley, A. K. (2000). Acid Phosphatases in the Haemolymph of the Desert Locust, Schistocerca gregaria, Infected with the Entomopathogenic Fungus Metarhizium anisopliae. Journal of Insect Physiology, 46, 1249-1257.