Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae)


  • Anđelina Radojević University of Belgrade - Faculty of Biology, 11000 Belgrade
  • Dejan Mirčić Department of Biomedical Sciences, State University of Novi Pazar, 36300 Novi Pazar
  • Miroslav Živić University of Belgrade - Faculty of Biology, 11000 Belgrade
  • Vesna Perić-Mataruga Institute for Biological Research “Siniša Stanković”, University of Belgrade, 11060 Belgrade
  • Milenka Božanić University of Belgrade - Faculty of Biology, 11000 Belgrade
  • Katarina Stojanović University of Belgrade - Faculty of Biology, 11000 Belgrade
  • Jovana Lukičić University of Belgrade - Faculty of Biology, 11000 Belgrade
  • Ivana Živić University of Belgrade - Faculty of Biology, 11000 Belgrade


antioxidant enzymes, biomarkers, Ecdyonurus venosus, organic pollution, oxidative stress


Paper description:

  • Molecular biomarkers of oxidative stress are the most apropriate parameters for detecting early influence of pollutants. The effects of trout farms on aquatic insects at the molecular level have been scantily investigated.
  • We examined the influence of trout farm effluents on the molecular biomarkers of oxidative stress in larvae of Ecdyonurus venosus as a very sensitive bioindicator of water quality.
  • Changes in components of antioxidative defense in E. venosus proved to be reliable and sensitive biochemical biomarkers of environmental stress caused by trout farm effluents, making this species an excellent bioindicator of freshwater pollution.

Abstract: The aim of the present study was to establish the extent to which the outlet waters of trout farms affect the ecosystems of the Crnica and Skrapež rivers in Serbia. We monitored selected biomarkers of oxidative stress: superoxide dismutase (SOD), glutathione peroxidase (GPx) and total glutathione (GSH) in larvae of the species Ecdyonurus venosus, and simultaneously analyzed the changes in the physical and chemical parameters. The investigations were carried out in spring at four localities along the Skrapež and Crnica rivers: one upstream (the control localities), and three downstream from the fish farm outlets. On the Skrapež River, the fish farm was clearly visible and was markedly changed by the chemical parameters of the water, manifested as a decreased concentration of dissolved oxygen, increased concentrations of nitrates, nitrites, ammonium ions, total phosphorus and total organic carbon, and increased SOD and GPx activities and decreased GSH concentration in larvae from the first downstream locality as compared to the control locality. On the Crnica River, due to the high values of water flow (around 3 m3/s), effluents from the fish farm had no effect on the chemical parameters of the water or on the tested biomarkers.

Received: December 20, 2018; Revised: January 29, 2019; Accepted: January 30, 2019; Published online: February 1, 2019

How to cite this article: Radojević A, Mirčić D, Živić M, Perić-Mataruga V, Božanić M, Stojanović K, Lukičić J, Živić I. Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae). Arch Biol Sci. 2019;71(2):225-33.


Download data is not yet available.


Søndergaard M, Jeppesen E. Anthropogenic impacts on lake and stream ecosystems, and approaches to restoration. J Appl Ecol. 2007;44:1089-94.

Benetti CJ, Pérez-Bilbao A, Garrido J. Macroinvertebrates as indicators of water quality in running waters: 10 years of research in rivers with different degrees of anthropogenic impacts. In: Voudouris K, editor. Ecological water quality - water treatment and reuse. Rijeka: InTech Press; 2012. p. 95-125.

Stojanović KZ. Uticaj pastrmskih ribnjaka na zajednice makrozoobentosa tekućica sa posebnim osvrtom na larve roda Baetis (Ephemeroptera, Insecta). [dissertation]. [Belgrade]: University of Belgrade, Faculty of Biology; 2017. 375 p.

Lin CK, Yi Y. Minimizing environmental impacts of freshwater aquaculture and reuse of pond effluents and mud. Aquaculture. 2003;226:57-68.

Pulatsu S, Rad F, Aksal G, Aydin F, Benil AC, Topcu A. The impact of rainbow trout farm effluents on water quality of Karasu stream, Turkey. Turk J Fish Aquat Sci. 2004;4:9-15.

Mirčić D, Stojanović K, Živić I, Todorović D, Stojanović D, Dolićanin Z, Perić-Mataruga V. The trout farm effect on Dinocras megacephala (Plecoptera: Perlidae) larvae: Antioxidative defense. Environ Toxicol Chem. 2016;35:1775-82.

Marković Z, Poleksić V, Mitrović Tutundžić V. Aquaculture in Serbia. In: Marković Z, editor. III International Conference “Fishery”; 2007 Feb 1-3; Belgrade, Serbia. Belgrade (Serbia): Faculty of Agriculture University of Belgrade; 2007. p. 35-40.

Marković Z, Poleksić V. Ribarstvo u Srbiji (Fishery in Serbia). Belgrade Serbia: Prof. dr Zoran Marković, Beograd, Srbija. 2011. 289 p. Serbian

Marković Z, Stanković M, Dulić Z, Živić I, Rašković B, Spasić M, Poleksić V. Aquaculture and fishery in Serbia-status and potentials. In: Marković Z, ed. V International conference Aquaculture & Fishery, 2011 Jun 1-3; Belgrade, Serbia. Belgrade (Serbia): Faculty of Agriculture University of Belgrade; 2011, p. 36-40.

Živić I, Marković Z, Filipović - Rojka Z, Živić M. Influence of a trout farm on water quality and macrozoobenthos communities of the receiving stream (Trešnjica River, Serbia). Int Rev Hydrobiol. 2009;94:673-87.

Soofiani NM, Hatami R, Hemami MR, Ebrahimi E. Effects of trout farm effluent on water quality and the macrobenthic invertebrate community of the Zayandeh-Roud River, Iran. North Amer J Aquacult. 2012;74:132-41.

Kripa PK, Prasanth KM, Sreejesh KK, Thomas TP. Aquatic macroinvertebrates as bioindicators of stream water quality - a case study in Koratty, Kerala, India. Res J Recent Sci. 2013;2:217-22.

Uherek CB, Gouveia, PBF. Biological monitoring using macroinvertebrates as bioindicators of water quality of Maroaga stream in the Maroaga Cave System, Presidente Figueiredo, Amazon, Brazil. Int J Ecol. 2014;2014: 308149.

Lock K, Goethals PLM. Distribution and ecology of the mayflies (Ephemeroptera) of Flanders (Belgium). Int J Limnol. 2011;47:159-65.

Landa V, Soldán T. Mayflies as bioindicators of water quality and environmental change on a regional and global scale. In: Corkum LD, Ciborowski JJH, editors. Current Directions in Research on Ephemeroptera. Toronto, Canada: Canadian Scholars' Press, Inc.; 1955. p. 21-9.

Alhejoj A, Salameh E, Bandel K. Mayflies (Order Ephemeroptera): an effective indicator of water bodies conditions in Jordan. IJSRES. 2014;2:361-70.

Winston GW, Di Giulio RT. Prooxidant and antioxidant mechanisms in aquatic organisms. Aquat Toxicol. 1991;19:137-61.

Valavanidis A, Vlahogianni T, Dassenakis M, Scoullos M. Molecular biomarkers of oxidative stress in aquatic organisms in relation to toxic environmental pollutants. Ecotoxicol Environ Saf. 2006;64:178-89.

Stoliar OB, Lushchak VI. Environmental pollution and oxidative stress in fish. In: Lushchak VI, editor. Oxidative stress - environmental induction and dietary antioxidants. Rijeka, Croatia: InTech,; 2012. p 132-66.

Vranković J, Labus-Blagojević S, Csanyi B, Makovinska J, Cvetković O, Gačić Z, Blagojević D, Paunović M. Antioxidant enzymes and GST activity in natural populations of Holandriana holandrii from the Bosna River. Turk J Biol. 2012;36:477-85.

Vranković J. Environmental impact on the antioxidant responses in Corbicula fluminea (Bivalvia: Veneroida: Corbiculidae) from the Danube River. Ital J Zool 2015;82:378-86.

Vranković J. Age-related changes in antioxidant and glutathione S-transferase enzyme activities in the Asian clam. Biochem (Mosc). 2016;81:339-49.

Lam PKS, Gray JS. The use of biomarkers in environmental monitoring programmes. Mar Pollut Bull. 2003;46:182-86.

Choi J. Biomarkers in environmental monitoring and its application in Chironomus spp. In: Hong SK, Lee JL, Ihm BS, Farina A, Son Y, Kim ES, editors. Ecological issues in a changing world - status, response and strategy. Dordrecht: Springer; 2004. p. 203-15.

Conti ME. Biomarkers for environmental monitoring. In: Conti ME, editor. Biological monitoring: Theory and applications. Wit Press; 2008; p. 25-46.

Hook SE, Gallagher EP, Batley GE. The Role of Biomarkers in the Assessment of Aquatic Ecosystem Health. Integr Environ Assess Manag. 2014;10:327-41.

Livingstone DR. Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Marine Poll Bull. 2001;42:656-66.

Livingstone DR. Oxidative stress in aquatic organisms in relation to pollution and aquaculture. Rev Med Vet. 2003;154:427-30.

Walsh A. Reference conditions and eutrophication impacts in Irish Rivers: Meeting the requirements of the water framework directive (2000-FS-2-M1): synthesis report. Wexford Ireland: Environmental Protection Agency. 2005; 353 p.

Borković SS, Šaponjić JS, Pavlović SZ, Blagojević DP, Milošević SM, Kovačević TB, Radojičić RM, Spasić MB, Žikić RV, Saičić ZS. The activity of antioxidant defence enzymes in mussel (Mytilus galloprovincialis) from the Adriatic Sea. Comp Biochem Physiol C Toxicol Pharmacol. 2005;141:366-74.

Šaponjić, JS, Borković SS, Kovačević TB, Pavlović SZ, Labus-Blagojević SD, Blagojević DP, Saičić ZS, Radojičić RM, Žikić RV, Spasić MB. Activity of antioxidant defense enzymes in Mediterranean sea shrimp (Parapenaeus longirostris): relation to the presence of PCBs and PAHs in the south Adriatic Sea. Period Biol. 108:2006;117-25.

Despotović SG, Perendija BR, Gavrić JP, Borković-Mitić SS, Paunović MM, Pavlović SZ, Sajčić ZS. Seasonal changes in oxidative stress biomarkers of the snail Viviparus acerosus from the Velika Morava river, Serbia. Arch Biol Sci. 2012;64:953-62.

Vranković Ј, Slavić M. Biomarker responses in Corbicula fluminea to the presence of dioxin-like polychlorinated biphenyls and seasonal changes. Ecol Indic. 2015;48:99-106.

Bjelanović K, Živić I, Dulić Z, Živić M, Đorđević J, Marinković S, Marković Z. Water quality assessment in the Raška river based on zoobenthos and zooplankton organisms as bioindicators. In: Poleksić V, ed. VI International conference Aquaculture & Fishery, 2013 Jun 12-14; Belgrade, Serbia. Belgrade (Serbia): Faculty of Agriculture University of Belgrade; 2013. p. 349-57.

Gavrilović Lj, Dukić D. Reke Srbije. Belgrade Serbia: Zavod za udžbenike i nastavna sredstva; 2002. 218 p. Serbian.

Bauernfeind E, Soldan T. The Mayflies of Europe (Ephemeroptera). Ollerup, Denmark: Apollo Books. 2012. 781 p.

Bradford MM. Arapid and sensitive method for the quantity of microgram quantities of protein utilizing the principle of protein - dye binding. Anal Biochem. 1976;72:248-54.

Misra HP, Fridovich I. The role of superoxide anion in the antioxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem. 1972;247:3170-175.

Tamura M, Oschino N, Chance B. Some characteristics of hydrogen and alkyl-hydroperoxides metabolizing systems in cardiac tissue. J Biochem. 1982;92:1019-31.

Griffith OW. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem. 1980;106:207-12.

Namin JI, Sharifinia M, Makrani AB. Assessment of fish farm effluents on macroinvertebrates based on biological indices in Tajan River (north Iran). CJES. 2013;11:29-39.

Boyd CE. Guidelines for aquaculture effluent management at the farm-level. Aquaculture. 2003;226:101-12.

Cho CY, Bureau DP. A review of diet formulaon strategies and feeding systems to reduce excretory and feed wastes in aquaculture. Aquacult Res. 2001;32:349-60.

Moraes MAB, Carmo CF, Tabata YA, Vaz-Dos-Santos AM, Mercante CTJ. Environmental indicators in effluent assessment of rainbow trout (Oncorhynchus mykiss) reared in raceway system through phosphorus and nitrogen. Braz J Biol. 2016;76:1021-8.

Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative Stress and Antioxidant Defense. World Allergy Organ J. 2012;5:9-19.

Shonouda ML, El-Samad LM, Mokhamer H, Toto N. Use of oxidative stress and genotoxic biomarkers of aquatic beetles Anaceana globulus (Coleoptera: Hydrophilidae) as biomonitors of water pollution. J Entomol. 2016;13:122-31.

Fridovich I. Superoxide anion radical (OFormula2), superoxide dismutases, and related matters. J Biol Chem. 1997;272:18515-7.

Łukaszewicz-Hussain A, Moniuszko-Jakoniuk J. Liver catalase, glutathione peroxidase and reductase activity, reduced glutathione and hydrogen peroxide levels in acute intoxication with chlorfenvinphos, an organophosphate insecticide. Pol J Environ Stud Polis. 2004;13:303-9.

Kelso BHL, Glass DM, Smith RV. Toxicity of nitrite in freshwater invertebrates. In: Wilson WS, Ball AS, Hinton RH, editors. Managing risks of nitrates to human and the environment. Cambridge: Royal Society of Chemistry; 1999. p. 175-88.

Alonso A, Camargo JA. Toxicity of nitrite to three species of freshwater invertebrate. Environ Toxicol. 2006;21:90-4.

Daniel TC, Sharpley AN, Lemunyon JL. Agricultural phosphorus and eutrophication: A symposium overview. J Environ Gual. 1998;27:251-7.




How to Cite

Radojević A, Mirčić D, Živić M, Perić-Mataruga V, Božanić M, Stojanović K, Lukičić J, Živić I. Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae). Arch Biol Sci [Internet]. 2019Jun.4 [cited 2024Feb.29];71(2):225-33. Available from: