The Use of Astacus leptodactylus (Eschscholtz, 1823) as a Test Species for Toxicity Evaluation of Municipal Wastewater Treatment Plant Effluents

Document Type : Original Research Paper


1 Department of Environmental Engineering, Faculty of Engineering, Munzur University, 62000 Tunceli, Turkey

2 Fisheries Faculty, Munzur University, TR62000, Tunceli, Turkey


This study evaluated the responses of biochemical biomarkers in Astacus leptodactylus exposed to treated municipal effluents discharged into Keban Dam Lake, Elazığ Elazig, Turkey. A. leptodatyclus were exposed to treated municipal effluents and Catalase (CAT), Superoxide dismutase (SOD) activity and lipid peroxidation (TBARS), glutathione (GSH) levels were measured as oxidative stress biomarkers.  SOD activity was increased after exposing to treated municipal effluents for 24th and 96th h. CAT activities were decreased from 25.29 to 14.12 nmol/min/ml compared to control in the group exposed to treated municipal effluents for 24 h but it increased after 96 h exposure. GSH levels were decreased from 9.08 to 3.77 µM compared to control, but MDA levels were increased both at 24 th h and 96 th h after exposure to treated municipal effluents. CAT, SOD activities and MDA and GSH levels in the hepatopancreas of A. leptodactylus are sensitive and suitable biochemical biomarkers for evaluating the toxicity of the treated municipal effluent complex mixtures. Treated municipal effluents exposure was found to cause sub-lethal responses in A. leptodactylus suggesting oxidative stress.


Aksu, O., Yildirim N.C., Yildirim, N., Danabas, D. and Danabas, S. (2015). Biochemical response of crayfish Astacus leptodactylus exposed to textile wastewater treated by indigenous white rot fungus Coriolus versicolor. Environ. Sci. Pollut. Res., 22:2987–2993.
Almroth, B.C., Albertsson, E., Sturve, J. and Forlin, L. (2008). Oxidative stress, evident in antioxidant defenses and damage products, in rainbow trout caged outside a sewage treatment plant. Ecotoxicol. Environ. Saf., 70:370–378.
Bainy, A.C.D., Saito, E., Carvalho, P.S.M. and Junqueira, V.B.C. (1996). Oxidative stress in gill, erythrocytes, liver and kidney of Nile tilapia (Oreochromis niloticus) from a polluted site. Aquat. Toxicol., 34: 151–162.
Barim, Ö. and Karatepe, M. (2010).The effects of pollution on the vitamins A, E, C, β-carotene contents and oxidative stress of the freshwater crayfish, Astacus leptodactylus. Ecotoxicol. Environ. Saf.,  73: 138-142.
Brix, H.  and Arias, C.A. (2005). The use of vertical flow constructed wetlands for on- site treatment of domestic wastewater: new Danish guidelines. Ecol. Eng., 25: 491–500. 
Cheesman, K.H. and Slater, T.F. (1993). Introduction to free radical biochemistry. Br. Med. Bull., 49: 481–493.
Chen, Y., Wang, C. and Wang, Z. (2005). Residues and source identification of persis- tent organic pollutants in farmland soils irrigated by effluents from biological treatment plants. Environ. Int., 31: 778–783.
Cristale, J. and Lacorte, S. (2015). PBDEs versus NBFR in wastewater treatment plants: occurrence and partitioning in water and sludge. AIMS Environ. Sci.,2: 533-546. 
Dautremepuits, C., Paris-Palacios, S. and  Betoulle, S. (2004). Modulation in hepatic and head kidney parameters of carp  (Cyprinus carpio L) induced by copper and chitosan. Comp. Biochem. Physiol C Toxicol. Pharmacol., 137:325–33. 
Diaz-Garduno, B., Perales, J.A., Biel-Maeso, M., Pintado-Herrera, M.G.,  Lara-Martin, P.A.,Garrido-Pérez, C. and Martín-Díaz, M.L. (2018). Biochemical responses of Solea senegalensis after continuous flow exposure to urban effluents. Sci. Total Environ., 615:486-497. 
Dimitrova, M.S.T., Tsinova, V. and Velcheva, V. (1994). Combined effect of zinc and lead on the hepatic superoxide dismutase–catalase system in carp (Cyprinus carpio). Comp. Biochem. Physiol.,108:43–6.
Fatima, R.A. and Ahmad, M. (2005). Certain antioxidant enzymes of Allium cepa as biomarkers for the detection of toxic heavy metals in wastewater. Sci. Total Environ., 346: 256-273.
Gagne,  F.,  Andre, C., Cejka, P., Hausler, R., Fournier, M. and Blaise, C. (2008). Immuno- toxic Effects on freshwater mussels of a primary-t,reated wastewater before and after ozonation:apilot plant study. Ecotoxicol. Environ. Saf., 69:366–373.
Geracitano, L.A., Monserrat, J.M. and Bianchini, A. (2004). Oxidative stress in Laeonereis acuta (Polychaeta, Nereididae):environmental and seasonal effects. Mar. Environ. Res., 58:625–630. 
Ghedira, J., Jebali, J., Bouraoui, Z., Banni, M., Chouba, L. and Boussetta, H. (2009). Acute Effects of Chlorpyriphos-ethyland Secondary Treated Effluents on Acetylcholinesterase and Butyryl Cholinesterase Activities in Carcinus maenas. J. Environ. Sci.,21:1–6.
Gillis, P.L., Gagné, F., McInnis, R., Hooey, T.M., Choy, E.S., André,  C., Hoque, M.E. and Metcalfe, C.D. (2014). The impact of municipal wastewater effluents on field-deployed freshwater mussels in the Grand River (ON). Environ. Toxicol. Chem., 33:134–143.
Harlioglu, M.M., Cakmak, M.N., Köprücü, K., Aksu, O., Harlioglu, A.G., Yonar, M., Duran, S.C.,Ozcan, S.T. and  Gundogdu, H. (2013). The effect of dietary n-3series fatty acids on the number of pleopadal egg and stage 1 juvenile in freshwater crayfish, Astacus leptodactylus Eschscholtz. Aquacult. Res., 44:860–868.
Karatas, F., Öbek, E. and Kamıslı, F. (2009). Antioxidant capacity of Lemna gibba L. exposed to wastewater treatment, Ecol. Eng., 35:1225–1230
Kamel, N., Jebali, J.,  Banni, M., Khedher, S.B., Chouba, L. and Boussetta, H. (2012).Biochemical responses and metals levels in Ruditapes decussatus after exposure to treated municipal effluents, Ecotoxicol. Environ. Saf., 82:40–46.
Kappus, H.A. (1987). A survey of chemicals inducing lipid peroxidation in biologicalSystems. Chem. Phys. Lipids 45: 105–115.
Machado, A.A.S., Wood, C.M., Bianchini, A. and Gillis, P.L. (2014). Responses of biomarkers in wild freshwater mussels chronically exposed to complex contaminant mixtures. Ecotoxicology 23:1345–1358.
Mehdi, H.H., Dickson, F., Leslie, M., Bragg, M., Servos, R. and Craig,  P.M. (2017). Impacts of wastewater treatment plant effluent on energetics and stress response of  rainbow darter (Etheostoma caeruleum) in the Grand River watershed. Comp. Biochem. Physiol. A,1096-4959:30184-7.
McCord, J.M. (1996). Effects of positive iron status at a cellular level. Nutr Rev 54:85–8
Monserrat. J,M,, Martınez. P.E., Geracitano, L.A., Amado, L.L., Martins, C.M., Pinho, G.L., Chaves, I.S., Ferreira-Cravo, M., Ventura-Lima, J. and Bianchini, A. (2007). Pollution biomarkers in estuarine animals: critical review and new perspectives. Comp. Biochem. Physiol. C, 146:221–234.
Movahedian, H., Bina, B. and Asghari, G.H. (2005).Toxicity Evaluation of Wastewater Treatment Plant Effluents Using Daphnia magna. Iranian J. Environ. Health Sci. Eng.,2: 1-4.
Noctor, G. and Foyer, C.H. (1998). Ascorbate and glutathione: keeping active oxygen undercontrol. Annu. Rev. Plant Physiol. Plant Mol. Biol., 49:249–279.  
Oakes, K.D. and VanDerKraak, G.J. (2003). Utility of the TBARS assay in detecting oxidative stress in whitesucker (Catostomus commersoni) populations exposed to  Pulpmil leffluent. Aquat. Toxicol., 63,447–463.
Petala, K.M., Samaras, L., Papadopoulos, P. and Zouboulis, A. (2009). Toxicological and ecotoxic impact of secondary and tertiary treated sewage effluents. Water Res., 43:5063–5074.
Prasad, M.N.V. (2004). Heavy Metal Stress in Plants: From Biomolecules to Ecosystems, Springer, Berlin Heidelberg Prasad,  K.V.S.K., Saradhi, P. and Sharmila. P, (1999), Concerted action of antioxidant enzymes and Curtailed growth under zinc toxicity in Brassica juncea. Environ. Exper. Bot., 42:1–10
Radic, S., Stipanicev, D., Cvjetko, P., Sırac,S., Pevalek-Kozlina, B. and Pavlica, M. (2011). Duckweed Lemna minor as a tool for testing  toxicity and genotoxicity of surface waters. Ecotoxicol. Environ. Saf., 74:182–187.
Sturve, J., Almroth, B.C. and Förlin, L. (2008). Oxidative stress in rainbow trout (Oncorhynchus mykiss) exposed to sewage treatment plant effluent. Ecotoxicol. Environ. Saf., 70: 446–452.
Upadhyay, A.R., Virendra, K., Mishra, V.K., Sudhir, K., Pandey, S.K. and Tripathi, B.D., (2007). Biofiltration of secondary treated municipal wastewater in a tropical city. Ecol. Eng., 30:9–15.
Tabrez, S. and Ahmad, M. (2009). Effect of wastewater intake on antioxidant and marker enzymes of tissue damage in rat tissues: Implications for the use of biochemical markers. Food Chem. Toxicol., 47:2465–78.
Tatar, S., Yildirim, N.C., Serdar, O., Yildirim, N. and Ogedey, A. (2018). The using of Gammarus pulex as a biomonitor in ecological risk assessment of secondary effluent from municipal wastewater treatment plant in Tunceli, Turkey. Hum. Ecol. Risk Assess, 24(3):819-829.
Tatar, S., Obek,  E. and Yildirim, N.C. (2017). Antioxidant Response in Duckweed After Exposure to Secondary Effluent from Municipal Wastewater Treatment Plant, Elazığ, Turkey. Bull. Environ. Contam. Toxicol., 99:399-404.
Valavanidis, A., Vlahogianni, T., Dassenakis, M. and Scoullos, M. (2006). Molecular biomarkers of oxidative stress in aquatic organisms in relation to  toxic environmental pollutants. Ecotoxicol. Environ. Saf., 64(2):178–189.
Vega-Lopez, A., Jimenez-Orozco, F.A., Garcia-Latorrec, E. and Dominguez-Lo´pez, M. (2008). Oxidative stress response in an endangered goodeidfish (Girardinichthys viviparus) by exposure to water from its extantlocalities. Ecotoxicol. Environ. Saf., 71:94–103.
Vioque-Fernandez, A., AlvesdeAlmeida, E. and Lo´pez-Barea, J. (2009). Assessment of Don˜ana National Park contamination in Procambarusclarkii: integration of conventional biomarkers and proteomic approaches. Sci. Total Environ., 407:1784–1797
Weis, J.S. and Weis, P. (2004). Metal uptake, transport and release by wetland plants: implications for phytoremediation and restoration. Environ. Int., 30:685–700.