Environmental Science and Pollution Research

, Volume 26, Issue 11, pp 10826–10834 | Cite as

Organ-specific responses to total ammonia nitrogen stress on juvenile grass carp (Ctenopharyngodon idellus)

  • Congcong Zhao
  • Jingtao XuEmail author
  • Xiaoli Xu
  • Qian Wang
  • Qiang Kong
  • Fei Xu
  • Yuanda Du
Research Article


Fish are important in constructed wetland (CW) ecosystem. An 80-day experiment was conducted by exposing juvenile grass carp (Ctenopharyngodon idellus) to 0, 0.5, 2.0, 4.5, 9.0, and 18.0 mg L−1 total ammonia nitrogen (TAN) stress to determine the severity of physiological changes in fish organs (liver, gills, and muscle) in CW. Specific growth rate results indicated that low TAN (≤ 2.0 mg L−1) help maintain or enhance grass carp growth. Fish physiological indexes did not significantly change during exposure, except for the gill’s reactive oxygen species (ROS) level that is susceptible to TAN exposure. Under high TAN (≥ 4.5 mg L−1), physiological changes and organ-specific responses were revealed. The ROS and malondialdehyde levels were higher in the gills than in the liver. At 9.0 mg L−1 TAN, the muscle cells manifested toxicity. The antioxidant system of different organs responded differently because the gills were more susceptible to low TAN than other organs. After TAN removal from the low TAN system, the antioxidative enzymes and antioxidants were increased to scavenge extra ROS and reverted to the normal level. However, grass carp cannot recover from the oxidative damage at ≥ 9.0 mg L−1 external TAN, resulting in organ dysfunction and failed ROS scavenging. This study provides information in maintaining CW sustainability.


Ammonia Grass carp Organ-specific Physiological response Constructed wetlands 


Funding information

This work was supported by the Natural Science Foundation of Shandong Province (ZR2016DB13) and the National Natural Science Foundation of China (21307078).


  1. Atle F, Albertk I, Bjørn R, Edward S, Sigurdo S (2009) Effects of chronic and periodic exposure to ammonia on growth and blood physiology in juvenile turbot (Scophthalmus maximus). Aquaculture 296(1):45–50Google Scholar
  2. Ballesteros ML, Rivetti NG, Morillo DO, Bertrand L, Amé MV, Bistoni MA (2017) Multi-biomarker responses in fish (Jenynsia multidentata) to assess the impact of pollution in rivers with mixtures of environmental contaminants. Sci Total Environ 595:711–722CrossRefGoogle Scholar
  3. Baudron AR, Needle CL, Rijnsdorp AD, Tara Marshall C (2014) Warming temperatures and smaller body sizes: synchronous changes in growth of North Sea fishes. Glob Chang Biol 20(4):1023–1031CrossRefGoogle Scholar
  4. Belarde TA, Railsback SF (2015) New predictions from old theory: emergent effects of multiple stressors in a model of piscivorous fish. Ecol Model 326:54–62CrossRefGoogle Scholar
  5. Boeuf G, Boujard D, Ruyet J (1999) Control of the somatic growth in turbot. J Fish Biol 55:128–147CrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1):248–254CrossRefGoogle Scholar
  7. Cheung CC, Zheng GJ, Li AM, Richardson BJ, Lam PK (2001) Relationships between tissue concentrations of polycyclic aromatic hydrocarbons and antioxidative responses of marine mussels, Perna viridis. Aquat Toxicol 52(3–4):189–203CrossRefGoogle Scholar
  8. Ching B, Chew SF, Wong WP, Ip YK (2009) Environmental ammonia exposure induces oxidative stress in gills and brain of Boleophthalmus boddarti (Mudskipper). Aquat Toxicol 95(3):203–212CrossRefGoogle Scholar
  9. Coban O, Kuschk P, Wells NS, Strauch G, Knoeller K (2014) Microbial nitrogen transformation in constructed wetlands treating contaminated groundwater. Environ Sci Pollut R, 1–11Google Scholar
  10. Deng Y, Wang W, Yu P, Xi Z, Xu L, Li X, He N (2013) Comparison of taurine, GABA, Glu, and Asp as scavengers of malondialdehyde in vitro and in vivo. Nanoscale Res Lett 8(1):1–9CrossRefGoogle Scholar
  11. Draper H, Hadley M (1990) A review of recent studies on the metabolism of exogenous and endogenous malondialdehyde. Xenobiotica 20(9):901–907CrossRefGoogle Scholar
  12. Dutra FM, Rönnau M, Sponchiado D, Forneck SC, Freire CA, Elc B (2017) Histological alterations in gills of Macrobrachium amazonicum juveniles exposed to ammonia and nitrite. Aquat Toxicol 187:115–123CrossRefGoogle Scholar
  13. Fernández D, García-Gómez C, Babín M (2013) In vitro evaluation of cellular responses induced by ZnO nanoparticles, zinc ions and bulk ZnO in fish cells. Sci Total Environ 452:262–274CrossRefGoogle Scholar
  14. Fonseca AR, Sanches Fernandes LF, Fontainhasfernandes A, Monteiro SM, Fal P (2017) The impact of freshwater metal concentrations on the severity of histopathological changes in fish gills: a statistical perspective. Sci Total Environ 599-600:217–226CrossRefGoogle Scholar
  15. Fu C, Cao ZD, Fu SJ (2013) The effects of caudal fin loss and regeneration on the swimming performance of three cyprinid fish species with different swimming capacities. J Eep Biol 216(16):3164–3174Google Scholar
  16. Gao N, Zhu L, Guo Z, Yi M, Zhang L (2017) Effects of chronic ammonia exposure on ammonia metabolism and excretion in marine medaka Oryzias melastigma. Fish Shellfish Immun 65:226–234CrossRefGoogle Scholar
  17. Halliwell B, Gutteridge J (1990) Role of free radical and catalytic metal ions in human disease: an overview. Methods Enzymol 186(186):1–85Google Scholar
  18. Hao L, Chen L (2012) Oxidative stress responses in different organs of carp (Cyprinus carpio) with exposure to ZnO nanoparticles. Ecotox Environ Safe 80(80):103–110CrossRefGoogle Scholar
  19. Ip Y, Chew S, Randall D (2001) Ammonia toxicity, tolerance, and excretion. Fish Physiol 20:109–148CrossRefGoogle Scholar
  20. Jayakumar AR, Rama Rao KV, Schousboe A, Norenberg MD (2004) Glutamine-induced free radical production in cultured astrocytes. Glia 46(3):296–301CrossRefGoogle Scholar
  21. Lin Y-C, Chen J-C, Man SNC, Morni WZW, Suhaili ASN, Cheng S-Y, Hsu C-H (2012) Modulation of innate immunity and gene expressions in white shrimp Litopenaeus vannamei following long-term starvation and re-feeding. Res Immunol 2:148–156CrossRefGoogle Scholar
  22. Mj DSS, Da CF, Leme FP, Takata R, Costa DC, Mattioli CC, Luz RK, Miranda-Filho KC (2017) Biological responses of Neotropical freshwater fish Lophiosilurus alexandri exposed to ammonia and nitrite. Sci Total Environ:616–617Google Scholar
  23. Pinto MR, Lucena MN, Faleiros RO, Almeida EA, McNamara JC, Leone FA (2016) Effects of ammonia stress in the Amazon River shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae). Aquat Toxicol 170:13–23CrossRefGoogle Scholar
  24. Rånby BG, Rabek JF (1978) Singlet oxygen reactions with organic compounds and polymers. John Wiley & SonsGoogle Scholar
  25. Randall DJ, Tsui TK (2002) Ammonia toxicity in fish. Mar Pollut Bull 45(1–12):17–23CrossRefGoogle Scholar
  26. Romano N, Zeng C (2009) Subchronic exposure to nitrite, potassium and their combination on survival, growth, total haemocyte count and gill structure of juvenile blue swimmer crabs, Portunus pelagicus. Ecotox Environ Safe 72(4):1287–1295CrossRefGoogle Scholar
  27. Romano N, Zeng C (2013) Toxic effects of ammonia, nitrite, and nitrate to decapod crustaceans: a review on factors influencing their toxicity, physiological consequences, and coping mechanisms. Rev Fish Sci 21(1):1–21CrossRefGoogle Scholar
  28. Simon TP (1998) Assessing the sustainability and biological integrity of water resources using fish communities. CRC PressGoogle Scholar
  29. Sinha AK, AbdElgawad H, Giblen T, Zinta G, De Rop M, Asard H, Blust R, De Boeck G (2014) Anti-oxidative defences are modulated differentially in three freshwater teleosts in response to ammonia-induced oxidative stress. PLoS One 9(4):e95319CrossRefGoogle Scholar
  30. Sinha AK, Zinta G, AbdElgawad H, Asard H, Blust R, De Boeck G (2015) High environmental ammonia elicits differential oxidative stress and antioxidant responses in five different organs of a model estuarine teleost (Dicentrarchus labrax). Comp Biochem Physiol Part C: Toxicol Pharmacol 174:21–31Google Scholar
  31. Stara A, Kristan J, Zuskova E, Velisek J (2012) Effect of chronic exposure to prometryne on oxidative stress and antioxidant response in common carp (Cyprinus carpio L.). Pestic Biochem Physiol 33(2): 334–343Google Scholar
  32. Sun H, Lü K, Minter EJ, Chen Y, Yang Z, Montagnes DJ (2012) Combined effects of ammonia and microcystin on survival, growth, antioxidant responses, and lipid peroxidation of bighead carp Hypophthalmythys nobilis larvae. J Hazard Mater 221-222(4):213–219CrossRefGoogle Scholar
  33. Sun H, Wang W, Li J, Yang Z (2014) Growth, oxidative stress responses, and gene transcription of juvenile bighead carp (Hypophthalmichthys nobilis) under chronic-term exposure of ammonia. Environ Toxicol Chem 33(8):1726–1731CrossRefGoogle Scholar
  34. Winzer K, Van Noorden CJ, Köhler A (2002) Glucose-6-phosphate dehydrogenase: the key to sex-related xenobiotic toxicity in hepatocytes of European flounder (Platichthys flesus L.)? Aquat Toxicol 56(4):275–288CrossRefGoogle Scholar
  35. Wood CM (2004) Dogmas and controversies in the handling of nitrogenous wastes: is exogenous ammonia a growth stimulant in fish? J Exp Biol 207(12):2043–2054CrossRefGoogle Scholar
  36. Xu J, Zhang J, Xie H, Li C, Bao N, Zhang C, Shi Q (2010) Physiological responses of Phragmites australis to wastewater with different chemical oxygen demands. Ecol Eng 36(10):1341–1347CrossRefGoogle Scholar
  37. Yin F, Peng S, Sun P, Shi Z (2011) Effects of low salinity on antioxidant enzymes activities in kidney and muscle of juvenile silver pomfret Pampus argenteus. Acta Ecol Sin 31(1):55–60CrossRefGoogle Scholar
  38. Yong Z, Hao-Ru T, Ya L (2008) Variation in antioxidant enzyme activities of two strawberry cultivars with short-term low temperature stress. World J Agric Sci 4(4):458–462Google Scholar
  39. Yu BP (1994) Cellular defenses against damage from reactive oxygen species. Hysiol Rev 74(1):139–162CrossRefGoogle Scholar
  40. Zhang J, Shen H, Wang X, Wu J, Xue Y (2004) Effects of chronic exposure of 2,4-dichlorophenol on the antioxidant system in liver of freshwater fish Carassius auratus. Chemosphere 55(2):167–174CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Congcong Zhao
    • 1
  • Jingtao Xu
    • 2
    Email author
  • Xiaoli Xu
    • 3
  • Qian Wang
    • 1
  • Qiang Kong
    • 1
  • Fei Xu
    • 1
  • Yuanda Du
    • 1
  1. 1.College of Geography and Environment, Collaborative Innovation Center of Human-Nature and Green Development in Universities of ShandongShandong Normal UniversityJinanPeople’s Republic of China
  2. 2.School of Municipal and Environmental EngineeringShandong Jianzhu UniversityJinanChina
  3. 3.School of Environmental Science and EngineeringShandong UniversityJinanChina

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