Penkonazole Maruz Bırakılan Tatlı Su Midyelerinin Solungaç Na+/K+-ATPaz, Mg2+-ATPaz, Ca2+-ATPaz ve Total-ATPaz Aktivitelerinin Değerlendirilmesi

Abstract

Triazol bir fungusit olan penkonazol, tarımsal üretimi arttırmak ve fungusları kontrol etmek için dünya çapında yaygın olarak kullanılmaktadır. Çift kabuklu yumuşakçalar, özellikle midyeler, sucul ekosistemlerde kirliliğin spesifik biyomonitor organizmalarıdır. Bu çalışmada tatlı su midyelerinin (Unio mancus) solungaç Na⁺/K⁺-ATPaz, Mg²⁺-ATPaz, Ca²⁺-ATPaz ve Total-ATPaz aktiviteleri üzerine penkonazol maruziyetinin olası toksik etkileri değerlendirilmiştir. Bu amaçla, 96 saatlik statik yenileme test sisteminde tatlı su midyeleri farklı konsantrasyonlarda penkonazole (1, 10, 100 ve 1000 µg AI L^-1) maruz bırakılmıştır. Toksisite testlerinde penkonazolün ticari formu (TOPAS^® 100 EC, Syngenta, Almanya) kullanılmıştır. Maruziyet ortamındaki gerçek penkonazol konsantrasyonları LC-MS/MS analizi ile belirlenmiştir. LC-MS/MS analizi sonuçlarına göre, maruziyet ortamındaki gerçek penkonazol konsantrasyonlarının, nominal penkonazol konsantrasyonlarından yaklaşık % 25 oranında daha düşük olduğu belirlenmiştir. 96 saatlik maruziyetten sonra, en yüksek penkonazol konsantrasyonu Mg²⁺-ATPaz hariç diğer ATPaz aktivitelerinin önemli düzeyde inhibisyonuna neden olmuştur (P < 0.05). Elde edilen veriler, akut penconazol maruziyetinin tatlı su midyeleri için potansiyel bir risk olabileceğini ve ATPaz enzimlerinin akuatik toksikoloji çalışmaları için önemli ve faydalı biyokimyasal belirteçler olduğunu göstermiştir.

Keywords

• Penkonazol toksisitesi
• Tatlı su midyesi
• ATPazlar
• LC-MS/MS

Assessment of Na+/K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, and Total-ATPase Activities in Gills of Freshwater Mussels Exposed to Penconazole

Abstract

Penconazole, a triazole fungicide, is widely used worldwide to increase agricultural production and to control fungus. Bivalves, especially mussels, are specific biomonitor organisms of pollution in aquatic ecosystems. This study deals with the possible toxic effects of exposure to penconazole on Na⁺/K⁺-ATPase, Mg²⁺-ATPase, Ca²⁺-ATPase, and Total-ATPase activities in the gills of freshwater mussels (Unio mancus). In this context, freshwater mussels were exposed to different concentrations of penconazole (1, 10, 100, and 1000 µg AI L^-1) in 96 hours static-renewal test system. The commercial form of penconazole (TOPAS^® 100 EC, Syngenta, Germany) was used in toxicity tests. The actual penconazole concentrations in the exposure media were determined by LC-MS/MS analysis. According to the LC-MS/MS analysis results, the actual penconazole concentrations in the exposure media were determined to be approximately 25% lower than the nominal penconazole concentrations. After 96 hours of exposure, the highest concentration of penconazole caused significant inhibition of ATPase activities except Mg²⁺-ATPase activity (P < 0.05). The observed data indicated that acute penconazole exposure may be a potential risk for freshwater mussels and that ATPase enzymes are important and useful biochemical markers for aquatic toxicology studies.

Keywords

• Penconazole toxicity
• Freshwater mussel
• ATPases
• LC-MS/MS

References

1. Aksakal, F. I., & Ciltas, A. (2018). Developmental toxicity of penconazole in Zebrafish (Danio rerio) embryos. Chemosphere, 200, 8-15.
2. Atkinson, A., Gatenby, A. D., & Lowe, A. G. (1973). The determination of inorganic orthophosphate in biological systems. Biochimica et Biophysica Acta (BBA)-General Subjects, 320(1), 195-204.
3. Atli, G., & Canli, M. (2011). Essential metal (Cu, Zn) exposures alter the activity of ATPases in gill, kidney and muscle of tilapia Oreochromis niloticus. Ecotoxicology, 20(8), 1861-1869.
4. Balasundaram, K., Ramalingam, K., & Selvarajan, V. R. (1995). Phosalone poisoning on the cation-linked ATPases of central nervous system of Rana tigrina (Daudin). Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology, 111(3), 451-455.
5. Begum, G. (2009). Enzymes as biomarkers of cypermethrin toxicity: response of Clarias batrachus tissues ATPase and glycogen phosphorylase as a function of exposure and recovery at sublethal level. Toxicology Mechanisms and Methods, 19(1), 29-39.
6. Begum, G. (2011). Organ-specific ATPase and phosphorylase enzyme activities in a food fish exposed to a carbamate insecticide and recovery response. Fish Physiology and Biochemistry, 37(1), 61-69.
7. Bradford, M., M. (1976). A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 72, 248–254.
8. Burgos-Aceves, M. A., & Faggio, C. (2017). An approach to the study of the immunity functions of bivalve haemocytes: physiology and molecular aspects. Fish & Shellfish Immunology, 67, 513-517.

9. Chaabane, M., Elwej, A., Ghorbel, I., Chelly, S., Mnif, H., Boudawara, T., Chaabouni, E. S., Zeghal, N., & Soudani, N. (2018). Penconazole alters redox status, cholinergic function and lung’s histoarchitecture of adult rats: Reversal effect of vitamin E. Biomedicine & Pharmacotherapy, 102, 645-652.
10. Chaabane, M., Tir, M., Hamdi, S., Boudawara, O., Jamoussi, K., Boudawara, T., Ghorbel, R.E., Zeghal, N., & Soudani, N. (2016). Improvement of heart redox states contributes to the beneficial effects of selenium against penconazole-induced cardiotoxicity in adult rats. Biological Trace Element Research, 169, 261-270.
11. Comoglio, L., Amin, O., Roque, A., Betancourt-Lozano, M., Anguas, D., & Haro, B. M. (2005). Evaluation of sublethal biomarkers in Litopenaeus vannamei on foodborne exposure to methyl parathion. Ecotoxicology and Environmental Safety, 62(1), 66-74.
12. David, M., Sangeetha, J., Harish, E., Shrinivas, J., & Naik, V. (2014). Deltamethrin induced alteration in Na+-K+, Mg2+, Ca2+ associated ATPases activity in the freshwarer fish Cirrhinus mrigala. International Journal of Pure and Applied Zoology, 2(2), 175-181.
13. Dogan, Z., Atli, G., & Canli, M. (2015). Effects of lead on ATPases in tissues of freshwater fish (Oreochromis niloticus) in differing calcium levels. Turkish Journal of Fisheries and Aquatic Sciences, 15(2), 223-233.
14. European Union (2006) Directive 2006/118/EC of the European Parliament and of the Council of 12 December 2006 on the protection of groundwater against pollution and deterioration. Official Journal of the European Union, 27/12/2006, L372/19.
15. European Union (2013). Directive 2013/39/EU of the European Parliament and of the Council of 12 August 2013 Amending Directives 2000/60/EC and 2008/105/EC as Regards Priority Substances in the Field of Water Policy. Official Journal of the European Union,24/8/2013, L226/1.
16. Guidi, P., Bernardeschi, M., Scarcelli, V., Cantafora, E., Benedetti, M., Falleni, A., & Frenzilli, G. (2017). Lysosomal, genetic and chromosomal damage in haemocytes of the freshwater bivalve (Unio pictorum) exposed to polluted sediments from the River Cecina (Italy). Chemistry and Ecology, 33(6), 516-527.
17. Husak, V.V., Mosiichuk, N.M., Storey, J.M., Storey, K.B., & Lushchak, V.I. (2017). Acute exposure to the penconazole-containing fungicide Topas partially augments antioxidant potential in goldfish tissues. Comparative Biochemistry and Physiology, Part C, 193, 1-8.
18. Iummato, M. M., Sabatini, S. E., Cacciatore, L. C., Cochón, A. C., Cataldo, D., de Molina, M. D. C. R., & Juárez, Á. B. (2018). Biochemical responses of the golden mussel Limnoperna fortunei under dietary glyphosate exposure. Ecotoxicology and Environmental Safety, 163, 69-75.
19. IUPAC, 2019. Environmental Fate - Ecotoxicology - Human Health - A to Z Index. Retrieved from https://sitem.herts.ac.uk/aeru/iupac/Reports/509.htm.
20. Juberg, D.R., Mudra, D.R., Hazelton, G.A., & Parkinson, A. (2006). The effect of fenbuconazole on cell proliferation and enzyme induction in the liver of female CD1 mice. Toxicology and Applied Pharmacology, 214, 178–187.
21. Korkmaz, V., Güngördü, A., & Ozmen, M. (2018). Comparative evaluation of toxicological effects and recovery patterns in zebrafish (Danio rerio) after exposure to phosalone-based and cypermethrin-based pesticides. Ecotoxicology and Environmental Safety, 160, 265-272.
22. Kulac, B., Atli, G., & Canli, M. (2013). Response of ATPases in the osmoregulatory tissues of freshwater fish Oreochromis niloticus exposed to copper in increased salinity. Fish Physiology and Biochemistry, 39(2), 391-401.
23. Lv, X., Pan, L.M., Wang, J.Y., Lu, L.P., Yan, W.L., Zhu, Y.Y., Xu, Y.W., Guo, M., & Zhuang, S.L. (2017). Effects of triazole fungicides on androgenic disruption and CYP3A4 enzyme activity. Environmental Pollution, 222, 504-512.
24. Nikolaou, S., Efstathiou, P., Tiggiridou, M., Arabatzis, N., Piera, Y., & Aletrari, M. (2017). Monitoring of Pesticides in Drinking, Surface and Ground Water of Cyprus by Liquid-Liquid and Solid Phase Extraction in Combination with GC/MS and UPLC/MS/MS. Journal of Water Resource and Protection, 9(10), 1184-1198.
25. Okay, O. S., Ozmen, M., Güngördü, A., Yılmaz, A., Yakan, S. D., Karacık, B., Tutak, B., & Schramm, K. W. (2016). Heavy metal pollution in sediments and mussels: assessment by using pollution indices and metallothionein levels. Environmental Monitoring Assessment, 188, 352.
26. Pałecz, D., Komuński, R., & Gabryelak, T. (2005). Na+/K+-ATPase activity as a biomarker of toxaphene toxicity in Unio tumidus. Toxicology in Vitro, 19(5), 707-712.
27. Parisi, M. G., Maisano, M., Cappello, T., Oliva, S., Mauceri, A., Toubiana, M., & Cammarata, M. (2019). Responses of marine mussel Mytilus galloprovincialis (Bivalvia: Mytilidae) after infection with the pathogen Vibrio splendidus. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 221, 1-9.
28. Parvez, S., Sayeed, I. and Raisuddin, S. (2006). Decreased Gill ATPase Activities in the Freshwater Fish Channa punctata (Bloch) Exposed to a Diluted Paper Mill Effluent. Ecotoxicology and Environmental Safety. 65, 62–66.
29. Peffer, R.C., Moggs, J.G., Pastoor, T., Currie, R.A., Wright, J., Milburn, G., Waechter, F., & Rusyn, I. (2007). Mouse liver effects of cyproconazole, a triazole fungicide: role of the constitutive androstane receptor. Toxicological Sciences, 99, 315–325.
30. Pham, B., Miranda, A., Allinson, G., & Nugegoda, D. (2017). Evaluating the non-lethal effects of organophosphorous and carbamate insecticides on the yabby (Cherax destructor) using cholinesterase (AChE, BChE), Glutathione S-Transferase and ATPase as biomarkers. Ecotoxicology and Environmental Safety, 143, 283-288.
31. Sancho, E., Fernandez-Vega, C., Ferrando, M. D., & Andreu-Moliner, E. (2003). Eel ATPase activity as biomarker of thiobencarb exposureç. Ecotoxicology and Environmental Safety, 56(3), 434-441.
32. Savorelli, F., Manfra, L., Croppo, M., Tornambè, A., Palazzi, D., Canepa, S., Trentini, P. L., Cicero, A. M., & Faggio, C. (2017). Fitness evaluation of Ruditapes philippinarum exposed to Ni. Biological Trace Element Research, 177(2), 384-393.
33. Saxena, T.B., Zacharıassen, K.E., & Jorgensen, L. (2000). Effects of Ethoxyquin on the Blood Composition of Turbot, Scophthalmus maximus L. Comparative Biochemistry and Physiology Part C. 127,1–9.
34. Sureda, A., Capó, X., Busquets-Cortés, C., & Tejada, S. (2018). Acute exposure to sunscreen containing titanium induces an adaptive response and oxidative stress in Mytillus galloprovincialis. Ecotoxicology and Environmental Safety, 149, 58-63.
35. Vieira, C. E. D., Pérez, M. R., Acayaba, R. D. A., Raimundo, C. C. M., & dos Reis Martinez, C. B. (2018). DNA damage and oxidative stress induced by imidacloprid exposure in different tissues of the Neotropical fish Prochilodus lineatus. Chemosphere, 195, 125-134.
36. Vijayavel, K., Gopalakrishnan, S., & Balasubramanian, M. P. (2007). Sublethal effect of silver and chromium in the green mussel Perna viridis with reference to alterations in oxygen uptake, filtration rate and membrane bound ATPase system as biomarkers. Chemosphere, 69(6), 979-986.
37. Wong, C.K.C., & Wong, M.H. (2000). Morphological and Biochemical Changes in the Gills of Tilapia (Oreochromis mossambicus) to Ambient Cadmium Exposure. Aquatic Toxicology. 48, 517–527.
38. Yang, D., Lu, X., Zhang, W., & He, F. (2002). Biochemical changes in primary culture of skeletal muscle cells following dimethoate exposure. Toxicology, 174, 79.
39. Zhang, X., Wang, X., Luo, F., Sheng, H., Zhou, L., Zhong, Q., Lou, Z., Sun, H., Yang, M., Cui, X., & Chen, Z. (2019). Application and enantioselective residue determination of chiral pesticide penconazole in grape, tea, aquatic vegetables and soil by ultra performance liquid chromatography-tandem mass spectrometry. Ecotoxicology and Environmental Safety, 172, 530-537.