, Volume 25, Issue 7, pp 1426–1437 | Cite as

Environmental concentration of carbamazepine accelerates fish embryonic development and disturbs larvae behavior

  • Liyuan Qiang
  • Jinping Cheng
  • Jun Yi
  • Jeanette M. Rotchell
  • Xiaotong Zhu
  • Junliang Zhou


Environmental pollution caused by pharmaceuticals has been recognized as a major threat to the aquatic ecosystems. Carbamazepine, as the widely prescribed antiepileptic drug, has been frequently detected in the aquatic environment and has created concerns about its potential impacts in the aquatic organisms. The effects of carbamazepine on zebrafish embryos were studied by examining their phenotype, behavior and molecular responses. The results showed that carbamazepine disturbed the normal growth and development of exposed zebrafish embryos and larvae. Upon exposure to carbamazepine at 1 μg/L, the hatching rate, body length, swim bladder appearance and yolk sac absorption rate were significantly increased. Embryos in treatment groups were more sensitive to touch and light stimulation. At molecular level, exposure to an environmentally relevant concentration (1 μg/L) of carbamazepine disturbed the expression pattern of neural-related genes of zebrafish embryos and larvae. This study suggests that the exposure of fish embryo to antiepileptic drugs, at environmentally relevant concentrations, affects their early development and impairs their behavior. Such impacts may have future repercussions by affecting fish population structure.


Carbamazepine Zebrafish larvae Accelerated development Behavior change Molecular response 



This study was supported by the Natural Science Foundation of Guangdong Province, China (No. s2012010010847), the New Century Excellent Researcher Award Program from Ministry of Education of China (No. NECT-12-0181), and the State Key Lab of Estuarine and Coastal Research (2012RCDW01).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.


  1. Ali S, Champagne DL, Alia A, Richardson MK (2011) Large-scale analysis of acute ethanol exposure in zebrafish development: a critical time window and resilience. PLoS One 6(5):e20037. doi: 10.1371/journal.pone.0020037 CrossRefGoogle Scholar
  2. Benotti MJ, Brownawell BJ (2007) Distributions of pharmaceuticals in an urban estuary during both dry-and wet-weather conditions. Environ Sci Technol 41(16):5795–5802CrossRefGoogle Scholar
  3. Brandão FP, Rodrigues S, Castro BB, Goncalves F, Antunes SC, Nunes B (2013) Short-term effects of neuroactive pharmaceutical drugs on a fish species: biochemical and behavioural effects. Aquat Toxicol 144–145:218–229CrossRefGoogle Scholar
  4. Bridges KN, Soulen BK, Overturf CL, Drevnick PE, Roberts AP (2015) Embryotoxicity of maternally-transferred methylmercury to fathead minnows (Pimephales promelas). Environ Toxicol Chem 9999(9999):1–6Google Scholar
  5. Brodin T, Fick J, Jonsson M, Klaminder J (2013) Dilute concentrations of a psychiatric drug alter behavior of fish from natural populations. Science 339(6121):814–815CrossRefGoogle Scholar
  6. Brun GL, Bernier M, Losier R, Doe K, Jackman P, Lee HB (2006) Pharmaceutically active compounds in Atlantic Canadian sewage treatment plant effluents and receiving waters, and potential for environmental effects as measured by acute and chronic aquatic toxicity. Environ Toxicol Chem 25(8):2163–2176CrossRefGoogle Scholar
  7. Calisto V, Domingues MRM, Erny GL, Esteves VI (2011) Direct photodegradation of carbamazepine followed by micellar electrokinetic chromatography and mass spectrometry. Water Res 45(3):1095–1104CrossRefGoogle Scholar
  8. CDC (Centers for Disease Control and Prevention) (2015) Chronic Disease Prevention and Health Promotion: targeting epilepsy. Accessed 27 Feb 2016
  9. Chen LG, Huang CJ, Hu CY, Yu K, Yang LH, Zhou BS (2012) Acute exposure to DE-71: effects on locomotor behavior and developmental neurotoxicity in zebrafish larvae. Environ Toxicol Chem 31(10):2338–2344CrossRefGoogle Scholar
  10. Colwill RM, Creton R (2011) Locomotor behaviors in zebrafish (Danio rerio) larvae. Behav Process 86:222–229CrossRefGoogle Scholar
  11. Cornell RA, Eisen JS (2002) Delta/Notch signaling promotes formation of zebrafish neural crest by repressing Neurogenin 1 function. Development 129(11):2639–2648Google Scholar
  12. Cueva-Mestanza R, Torres-Padrón ME, Sosa-Ferrera Z, Santana-Rodríguez JJ (2008) Microwave-assisted micellar extraction coupled with solidphase extraction for preconcentration of pharmaceuticals in molluscs prior to determination by HPLC. Biomed Chromatogr 22(10):1115–1122CrossRefGoogle Scholar
  13. de Jongh CM, Kooij PJF, de Voogt P, ter Laak TL (2012) Screening and human health risk assessment of pharmaceuticals and their transformation products in Dutch surface waters and drinking water. Sci Total Environ 427–428(15):70–77CrossRefGoogle Scholar
  14. Delgado LF, Charles P, Glucina K, Morlay C (2012) QSAR-like models: a potential tool for the selection of PhACs and EDCs for monitoring purposes in drinking water treatment systems—A review. Water Res 46(19):6196–6209CrossRefGoogle Scholar
  15. Fent K, Weston AA, Caminada D (2006) Ecotoxicology of human pharmaceuticals. Aquat Toxicol 76(2):122–159CrossRefGoogle Scholar
  16. Galus M, Rangarajan S, Lai A, Shaya L, Balshine S, Wilson JY (2014) Effects of chronic, parental pharmaceutical exposure on zebrafish (Danio rerio) offspring. Aquat Toxicol 151:124–134CrossRefGoogle Scholar
  17. Greaney NE, Mannion KL, Dzieweczynski TL (2015) Signaling on Prozac: altered audience effects on male-male interactions after fluoxetine exposure in Siamese fighting fish. Behav Ecol Sociobiol 69(12):1925–1932CrossRefGoogle Scholar
  18. Gros M, Petrovi´c M, Barcel´o D (2006) Development of a multi-residue analytical methodology based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) for screening and trace level determination of pharmaceuticals in surface and wastewaters. Talanta 70(4):678–690CrossRefGoogle Scholar
  19. Grover DP, Zhou JL, Frickers PE, Readman JW (2011) Improved removal of estrogenic and pharmaceutical compounds in sewage effluent by full scale granular activated carbon: impact on receiving river water. J Hazard Mater 185(2–3):1005–1011CrossRefGoogle Scholar
  20. Hao C, Lissemore L, Nguyen B, Kleywegt S, Yang P, Solomon K (2006) Determination of pharmaceuticals in environmental waters by liquid chromatography/electrospray ionization/tandem mass spectrometry. Anal Bioanal Chem 384(2):505–513CrossRefGoogle Scholar
  21. Harkin G, Hopkinson H (2010) Carbamazepin Pract Diab Int 27(5):205–206CrossRefGoogle Scholar
  22. Huerta-Fontela M, Galceran MT, Ventura F (2011) Occurrence and removal of pharmaceuticals and hormones through drinking water treatment. Water Res 45(3):1432–1442CrossRefGoogle Scholar
  23. Jeong JY, Einhorn Z, Mercurio S, Lee S, Lau B, Mione M, Wilson SW, Guo S (2006) Neurogenin1 is a determinant of zebrafish basal forebrain dopaminergic neurons and is regulated by the conserved zinc finger protein Tof/Fezl. Proc Natl Acad Sci USA 103(13):5143–5148CrossRefGoogle Scholar
  24. Kleywegt S, Pileggi V, Yang P, Hao C, Zhao XM, Rocks C, Thach S, Cheung P, Whitehead B (2011) Pharmaceuticals, hormones and bisphenol A in untreated source and finished drinking water in Ontario, Canada—Occurrence and treatment efficiency. Sci Total Environ 409(8):1481–1488CrossRefGoogle Scholar
  25. Kokel D, Peterson RT (2011) Using the zebrafish photomotor response for psychotropic drug screening. Methods Cell Biol 105:517–524CrossRefGoogle Scholar
  26. Komesli OT, Muz M, Ak MS, Bakırdere S, Gokcay CF (2015) Occurrence, fate and removal of endocrine disrupting compounds (EDCs)in Turkish wastewater treatment plants. Chem Eng J 277:202–208CrossRefGoogle Scholar
  27. Kyzar EJ, Collins C, Gaikwad S, Green J, Roth A, Monnig L, El-Ounsi M, Davis A, Freeman A, Capezio N, Stewart AM, Kalueff AV (2012) Effects of hallucinogenic agents mescaline and phencyclidine on zebrafish behavior and physiology. Prog Neuro-Psychoph 37:194–202CrossRefGoogle Scholar
  28. Liu C, Wen XW, Ge Y, Chen N, Hu WH, Zhang T, Zhang JG, Meng FG (2013) Responsive neurostimulation for the treatment of medically intractable epilepsy. Brain Res Bull 97:39–47CrossRefGoogle Scholar
  29. Ma Q, Kintner C, Anderson DJ (1996) Identification of neurogenin, a vertebrate neuronal determination gene. Cell 87(1):43–52CrossRefGoogle Scholar
  30. Mailler R, Gasperi J, Coquet Y, Deshayes S, Zedek S, Cren-Olive C, Cartiser N, Eudes V, Bressy A, Caupos E, Moilleron R, Chebbo G, Rocher V (2015) Study of a large scale powdered activated carbon pilot: removals of a wide range of emerging and priority micropollutants from wastewater treatment plant effluents. Water Res 72:315–330CrossRefGoogle Scholar
  31. Maximino C, Puty B, Benzecry R, Araújo J, Lima MG, Batista EDJO, Oliveira KRDM, Crespo-Lopez ME, Herculano AM (2013) Role of serotonin in zebrafish (Danio rerio) anxiety: relationship with serotonin levels and effect of buspirone, WAY 100635, SB 224289, fluoxetine and para-chlorophenylalanine (pCPA) in two behavioral models. Neuropharmacology 71:83–97CrossRefGoogle Scholar
  32. McCann SM, Rettori V (1986) Gamma amino butyric acid (GABA) controls anterior pituitary hormone secretion. Adv Biochem Psychopharmacol 42:173–189Google Scholar
  33. McEneff G, Barron L, Kelleher B, Paull B, Quinn B (2013) The determination of pharmaceutical residues in cooked and uncooked marine bivalves using pressurized liquid extraction, solid-phase extraction and liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem 405(29):9509–9521CrossRefGoogle Scholar
  34. Miao XS, Yang JJ, Metcalfe CD (2005) Carbamazepine and its metabolites in wastewater and in biosolids in a municipal wastewater treatment plant. Environ Sci Technol 39(19):7469–7475CrossRefGoogle Scholar
  35. Mittelbach GG, Ballew NG, Kjelvik MK (2014) Fish behavioral types and their ecological consequences. Can J Fish Aquat Sci 71(6):927–944CrossRefGoogle Scholar
  36. Miyares RL, de Rezende VB, Farber SA (2004) Zebrafish yolk lipid processing: a tractable tool for the study of vertebrate lipid transport and metabolism. Dis Model Mech 7(7):915–927CrossRefGoogle Scholar
  37. Moshé SL, Perucca E, Ryvlin P, Tomson T (2015) Epilepsy: new advances. Lancet 385(9971):884–898CrossRefGoogle Scholar
  38. Nassef M, Matsumoto S, Seki M, Khalil F, Kang IJ, Shimasaki Y, Oshima Y, Honjo T (2010) Acute effects of triclosan, diclofenac and carbamazepine on feeding performance of Japanese medaka fish (Oryzias latipes). Chemosphere 80(9):1095–1100CrossRefGoogle Scholar
  39. Oertel WH, Mugnaini E, Tappaz ML, Weise VK, Dahl AL, Schmechel DE, Kopin AI (1982) Central GABAergic innervation of neurointermediate pituitary lobe: biochemical and immunocytochemical study in the rat. Proc Natl Acad Sci USA 79:675–679CrossRefGoogle Scholar
  40. Padhye LP, Yao H, Kung’u FT, Huang CH (2014) Year-long evaluation on the occurrence and fate of pharmaceuticals, personal care products, and endocrine disrupting chemicals in an urban drinking water treatment plant. Water Res 51:266–276CrossRefGoogle Scholar
  41. Palmer PM, Wilson LR, O’Keefe P, Sheridan R, King T, Chen CY (2008) Sources of pharmaceutical pollution in the New York City Watershed. Sci Total Environ 394(1):90–102CrossRefGoogle Scholar
  42. Pittman JT, Lott CS (2014) Startle response memory and hippocampal changes in adult zebrafish pharmacologically-induced to exhibit anxiety/depression-like behaviors. Physiol Behav 123:174–179CrossRefGoogle Scholar
  43. Rabiet M, Togola A, Brissaud F, Seidel JL, Budzinski H, Elbaz-Poulichet F (2006) Consequences of treated water recycling as regards pharmaceuticals and drugs in surface and ground waters of a medium-sized Mediterranean catchment. Environ Sci Technol 40(17):5282–5288CrossRefGoogle Scholar
  44. Ramirez AJ, Brain RA, Usenko S, Mottaleb MA, O’Donnell JG, Stahl LL, Wathen JB, Snyder BD, Pitt JL, Perez-Hurtado P, Dobbins LL, Brooks BW, Chambliss CK (2009) Occurrence of pharmaceutical and personal care products in fish: results of a national pilot study in the United States. Environ Toxicol Chem 28(12):2587–2597CrossRefGoogle Scholar
  45. Rinkwitz S, Mourrain P, Becker TS (2011) Zebrafish: an integrative system for neurogenomics and neurosciences. Prog Neurobiol 93(2):231–243CrossRefGoogle Scholar
  46. Shenker M, Harush D, Ben-Ari J, Chefetz B (2011) Uptake of carbamazepine by cucumber plants—a case study related to irrigation with reclaimed wastewater. Chemosphere 82(6):905–910CrossRefGoogle Scholar
  47. Smith BR, Blumstein DT (2008) Fitness consequences of personality: a meta-analysis. Behav Ecol 19(2):448–455CrossRefGoogle Scholar
  48. Strecker R. (2013) Toxicity and teratogenesis in zebrafish embryos (Danio rerio). Dissertation, Ruperto-Carola University of HeidelbergGoogle Scholar
  49. Valcárcel Y, Alonso SG, Rodríguez-Gil JL, Castaño A, Montero JC, Criado-Alvarez JJ, Mirón IJ, Catalá M (2013) Seasonal variation of pharmaceutically active compounds in surface (Tagus River) and tap water (Central Spain). Environ Sci Pollut R 20(3):1396–1412CrossRefGoogle Scholar
  50. van den Brandhof EJ, Montforts M (2010) Fish embryo toxicity of carbamazepine, diclofenac and metoprolol. Ecotox Environ Safe 73(8):1862–1866CrossRefGoogle Scholar
  51. van Woudenberg AB, Snel C, Rijkmans E, de Groot D, Bouma M, Hermsen S, Piersma A, Menke A, Wolterbeek A (2014) Zebrafish embryotoxicity test for developmental (neuro) toxicity: demo case of an integrated screening approach system using anti-epileptic drugs. Reprod Toxicol 49:101–116CrossRefGoogle Scholar
  52. Vulliet E, Cren-Olive´ C, Grenier-Loustalot MF (2011) Occurrence of pharma-ceuticals and hormones in drinking water treated from surface waters. Environ Chem Lett 9(1):103–114CrossRefGoogle Scholar
  53. Wang SL, Zhou N (2016) Removal of carbamazepine from aqueous solution using sono-activated persulfate process. Ultrason Sonochem 29:156–162CrossRefGoogle Scholar
  54. Wang C, Shi HL, Adams CD, Gamagedara S, Stayton I, Timmons T, Ma YF (2011) Investigation of pharmaceuticals in Missouri natural and drinking water using high performance liquid chromatography-tandem mass spectrometry. Water Res 45(4):1818–1828CrossRefGoogle Scholar
  55. Westerfield M (2000) The zebrafish book: A guide for the laboratory use of zebrafish (Danio rerio). Eugene (Oregon)Google Scholar
  56. WHO (World Health Organization) (2016) Media centre: epilepsy. Accessed 27 February 2016
  57. Wiegel S, Aulinger A, Brockmeyer R, Harms H, Loffler J, Reincke H, Schmidt R, Stachel B, von Tumpling W, Wanke A (2004) Pharmaceuticals in the river Elbe and its tributaries. Chemosphere 57(2):107–126CrossRefGoogle Scholar
  58. Wille K, Kiebooms JAL, Claessens M, Rappé K, Bussche JV, Noppe H, Praet NV, Wulf ED, Caeter PV, Janssen CR, Brabander HFD, Vanhaecke L (2011) Development of analytical strategies using U-HPLC-MS/MS and LC-ToF-MS for the quantification of micropollutants in marine organisms. Anal Bioanal Chem 400(5):1459–1472CrossRefGoogle Scholar
  59. Xin Q, Rotchell JM, Cheng JP, Yi J, Zhang Q (2015) Silver nanoparticles affect the neural development of zebrafish embryos. J Appl Toxicol 35(12):1481–1492CrossRefGoogle Scholar
  60. Yan Q, Zhang YX, Kang J, Gan XM, Xu-Y P, Guo JS, Gao X (2015) A preliminary study on the occurrence of pharmaceutically active compounds in the river basins and their removal in two conventional drinking water treatment plants in Chongqing, China. CLEAN–Soil, Air. Water 43(6):794–803Google Scholar
  61. Zhang Y, Geißen SU, Gal C (2008) Carbamazepine and diclofenac: removal in wastewater treatment plants and occurrence in water bodies. Chemosphere 73(8):1151–1161CrossRefGoogle Scholar
  62. Zhang Q, Cheng JP, Xin Q (2015) Effects of tetracycline on developmental toxicity and molecular responses in zebrafish (Danio rerio) embryos. Ecotoxicology 24(4):707–719CrossRefGoogle Scholar
  63. Zhou XF, Dai CM, Zhang YL, Surampalli RY, Zhang TC (2011) A preliminary study on the occurrence and behavior of carbamazepine (CBZ) in aquatic environment of Yangtze River Delta. China Environ Monit Assess 173(1–4):45–53CrossRefGoogle Scholar
  64. Zuehlke S, Duennbier U, Heberer T (2004) Determination of polar drug residues in sewage and surface water applying liquid chromatography-tandem mass spectrometry. Anal Chem 76(22):6548–6554CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Liyuan Qiang
    • 1
  • Jinping Cheng
    • 1
    • 2
  • Jun Yi
    • 1
  • Jeanette M. Rotchell
    • 3
  • Xiaotong Zhu
    • 1
  • Junliang Zhou
    • 1
    • 4
  1. 1.State Key Laboratory of Estuarine and Coastal ResearchEast China Normal UniversityShanghaiChina
  2. 2.Environmental Science Programs, School of ScienceHong Kong University of Science and TechnologyKowloonChina
  3. 3.School of Biological, Biomedical & Environmental SciencesUniversity of HullHullUK
  4. 4.School of Civil and Environmental EngineeringUniversity of Technology SydneyBroadwayAustralia

Personalised recommendations