The Effect of Bisphenol A on Growth, Morphology, Lipid Peroxidation, Antioxidant Enzyme Activity, and PS II in Cylindrospermopsis raciborskii and Scenedesmus quadricauda

  • Rong Xiang
  • Junqiong Shi
  • Yi Yu
  • Hongbo Zhang
  • Congcong Dong
  • Yanjun Yang
  • Zhongxing Wu


To investigate the effect of bisphenol A (BPA) on Cylindrospermopsis raciborskii (Cyanobacteria) and Scenedesmus quadricauda (Chlorophyta), we grew the two species at BPA concentrations of 0, 0.1, 1, 2, 5, 10, and 20 mg/L and examined their growth, lipid peroxidation, antioxidant enzyme activity, and chlorophyll a fluorescence. The 96-h EC50 values (effective concentration causing 50% growth inhibition) for BPA in C. raciborskii and S. quadricauda were 9.663 ± 0.047, and 13.233 ± 0.069 mg/L, respectively. A significant reduction in chlorophyll a concentration was found in C. raciborskii and S. quadricauda when BPA concentrations were greater than 1 and 2 mg/L, respectively. Furthermore, F v/F m, ΔF/F m′, and qP decreased significantly at 10 mg/L BPA in C. raciborskii but started to decrease at 10 mg/L in S. quadricauda. The changes in chlorophyll fluorescence parameters (α, rETRmax) that were obtained from the rapid light response curves of both algae species showed similar responses to F v/F m, ΔF/F m′, and qP under BPA-induced stress. Values for all of the chlorophyll fluorescence parameters in S. quadricauda were higher than in C. raciborskii; however, the nonphotochemical quenching measured in C. raciborskii was considerably higher than it was in S. quadricauda. In addition, lipid peroxidation (determined as MDA content) and antioxidant enzyme activities (SOD and CAT) increased in both species as the BPA concentration increased. These results suggest that C. raciborskii is more sensitive to the effects of BPA than S. quadricauda and that photosystem II might be a target for the activity of BPA in vivo.



This research was supported by Fundamental Research Funds for the Central Universities (XDJK2016C111, XDJK2017B010) and Natural Science Foundation Project of China SWU (SWNUB2011).


  1. Abdel-Hamid MI (1996) Development and application of a simple procedure for toxicity testing using immobilized algae. Water Sci Technol 33:129–138Google Scholar
  2. Adams III WW, Demmig-Adams B (2004) Chlorophyll fluorescence as a tool to monitor plant response to the environment. In: Papageorgiou GC, Govindjee (eds) Chloropyll fluorescence: a signature of photosynthesis. Springer, Netherlands, pp 583–604Google Scholar
  3. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefGoogle Scholar
  4. Alexander HC, Dill DC, Smith LW, Guiney PD, Dorn P (1988) Bisphenol a: acute aquatic toxicity. Environ Toxicol Chem 7:19–26CrossRefGoogle Scholar
  5. Ali I, Liu B, Farooq MA, Islam F, Azizullah A (2015) Toxicological effects of bisphenol A on growth and antioxidant defense system in Oryza sativa as revealed by ultrastructure analysis. Mandarin 124:277–284Google Scholar
  6. Ali I, Jan M, Wakeel A, Azizullah A, Liu B (2017) Biochemical responses and ultrastructural changes in ethylene insensitive mutants of Arabidopsis thialiana subjected to bisphenol A exposure. Ecotoxicol Environ Saf 144:62–71CrossRefGoogle Scholar
  7. An S, Mo Y, Ou C (2002) Probit analysis with SPSS 10.0 software. J First Mil Med Univ 22:1019–1022 (in Chinese) Google Scholar
  8. Auriol M, Filali-Meknassi Y, Tyagi RD, Adams CD, Rao YS (2006) Endocrine disrupting compounds removal from wastewater, a new challenge. Process Biochem 41:525–539CrossRefGoogle Scholar
  9. Baehrs H, Putschew A, Steinberg CEW (2013) Toxicity of hydroquinone to different freshwater phototrophs is;influenced by time of exposure and pH. Environ Sci Pollut Res 20:146–154CrossRefGoogle Scholar
  10. Bai F, Liu R, Yang Y, Ran X, Shi J, Wu Z (2014) Dissolved organic phosphorus use by the invasive freshwater diazotroph cyanobacterium, Cylindrospermopsis raciborskii. Harmful Algae 39:112–120CrossRefGoogle Scholar
  11. Baumann HA, Morrison L, Stengel DB (2009) Metal accumulation and toxicity measured by PAM-Chlorophyll fluorescencein seven species of marine macroalgae. Ecotoxicol Environ Saf 72:1063–1075CrossRefGoogle Scholar
  12. Birnbaum LS, Fenton SE (2003) Cancer and developmental exposure to endocrine disruptors. Environ Health Perspect 111:389–394CrossRefGoogle Scholar
  13. Bodin J, Bolling A, Samuelsen M, Becher R, Lovik M, Nygaard U (2013) Long-term bisphenol A exposureaccelerates insulitis development in diabetes-prone NOD mice. Immunopharmacol Immunotoxicol 35:349–358CrossRefGoogle Scholar
  14. Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  15. Calatayud A, Barreno E (2001) Chlorophyll a fluorescence, antioxidant enzymes and lipid peroxidation in tomato in response to ozone and benomyl. Environ Pollut 115:283–289CrossRefGoogle Scholar
  16. Chen HX, Gao HY, An S, Li WJ (2004) Dissipation of excess energy in Mehler-Peroxidase reaction in Rumex leaves during salt shock. Photosynthetica 42:117–122CrossRefGoogle Scholar
  17. Choo KS, Snoeijs P, Pedersén M (2004) Oxidative stress tolerance in the filamentous green algae Cladophora glomerata and Enteromorpha ahlneriana. J Exp Mar Biol Ecol 298:111–123CrossRefGoogle Scholar
  18. Comerton AM, Andrews RC, Bagley DM, Yang P (2007) Membrane adsorption of endocrine disrupting compounds and pharmaceutically active compounds. J Membr Sci 303:267–277CrossRefGoogle Scholar
  19. Dhindsa RS, Matowe W (1981) Drought tolerance in two mosses: with enzymatic defense against lipid peroxidation. J Exp Bot 32:79–91CrossRefGoogle Scholar
  20. Dodds EC, Lawson W (1936) Synthetic estrogenic agents without the phenanthrene nucleus. Nature 137:996CrossRefGoogle Scholar
  21. Dosnon-Olette R, Trotel-Aziz P, Couderchet M, Eullaffroy P (2010) Fungicides and herbicide removal in Scenedesmus cell suspensions. Chemosphere 79:117–123CrossRefGoogle Scholar
  22. Dyble J, Paer HW, Neilan BA (2002) Genetic characterization of Cylindrospermopsis raciborskii (Cyanobacteria) isolates from diverse geographic origins based on nifH and cpcBA-IGS nucleotide sequence analysis. Appl Environ Microbiol 68:2567–2571CrossRefGoogle Scholar
  23. Ebenezer V, Ki JS (2016) Toxic effects of Aroclor 1016 and bisphenol A on marine green algae Tetraselmis suecica, diatom Ditylum brightwellii and dinoflagellate Prorocentrum minimum. Korean J Microbiol 52:306–312CrossRefGoogle Scholar
  24. Fromme H, Küchler Otto T, Pilz K, Müller J (2002) Occurrence of phthalates and bisphenol A and F in the environment. Water Res 36:1429–1438CrossRefGoogle Scholar
  25. Gao Y, Cui Y, Xiong W, Li X, Wu Q (2010) Effect of UV-C on algal evolution and differences in growth rate, pigmentation and photosynthesis between prokaryotic and eukaryotic algae. Photochem Photobiol 85:774–782CrossRefGoogle Scholar
  26. Gassman NR (2017) Induction of oxidative stress by bisphenol A and its pleiotropic effects. Environ Mol Mutagen 58:60–71CrossRefGoogle Scholar
  27. Gattullo CE, Bährs H, Steinberg CEW, Loffredo E (2012) Removal of bisphenol A by the freshwater green alga Monoraphidium braunii and the role of natural organic matter. Sci Total Environ 416:501–506CrossRefGoogle Scholar
  28. Geoffroy L, Dewez D, Vernet G, Popovic R (2003) Oxyfluorfen toxic effect on Scenedesmus obliquus evaluated by different photosynthetic and enzymatic biomarkers. Arch Environ Contam Toxicol 45:445–452CrossRefGoogle Scholar
  29. Guo R, Ebenezer V, Ki JS (2012) Transcriptional responses of heat shock protein 70 (Hsp70) to thermal, bisphenol A, and copper stresses in the dinoflagellate Prorocentrum minimum. Chemosphere 89:512–520CrossRefGoogle Scholar
  30. Halliwell B, Buzadzić B, Spasic M, Saicić ZS, Saicić R (1990) Antioxidant defenses in the ground squirrel Citellus citellus. 1. A comparison with the rat. Free Radic Biol Med 9:401–406CrossRefGoogle Scholar
  31. Hamilton MA, Russo RC, Thurston RV (1977) Thurston trimmed Spearman-Karber method for estimating median lethal concen-trations in toxicity bioassays. Environ Sci Technol 11:714–719CrossRefGoogle Scholar
  32. He J, Chee CW, Goh CJ (1996) “Photoinhibition” of Heliconia under natural tropical conditions: the importance of leaf orientation for light interception and leaf temperature. Plant Cell Environ 19:1238–1248CrossRefGoogle Scholar
  33. Hecky RE, Kilham P (1988) Nutrient limitation of phytoplankton in freshwater and marine environments: a review of recent evidence on the effects of enrichment. Limnol Oceanogr 33:796–822Google Scholar
  34. Henley WJ (1993) On the measurement and interpretation of photosynthetic light-response curves in algae in the context of photoinhibition and diel changes. J Phycol 29:729–739CrossRefGoogle Scholar
  35. Hoekstra EJ, Simoneau C (2013) Release of bisphenol A from polycarbonate-are view. Crit Rev Food Sci Nutr 3:386–402CrossRefGoogle Scholar
  36. Ichimura T (1979) Isolation and culture methods of algae. KyorituShuppan, Tokyo. (In Japanese without English title). pp 294–305Google Scholar
  37. Kavlock R (1999) Overview of endocrine disruptor research activity in the United States. Chemosphere 39:1227–1236CrossRefGoogle Scholar
  38. Kim K, Park H (2013) Association between urinary concentration of bisphenol A and type 2 diabetes in Korean adults: a population-based cross-sectional study. Int J Hyg Environ Health 216:467–471CrossRefGoogle Scholar
  39. Kim HS, Han SY, Yoo SD et al (2001) Potential estrogenic effects of bisphenol-A estimated by in vitro and in vivo combination assays. Toxicol Sci 26:111–118CrossRefGoogle Scholar
  40. Kortenkamp A (2007) Ten years of mixing cocktails: a review of combination effects of endocrine-disrupting chemicals. Environ Health Perspect 115:98–103CrossRefGoogle Scholar
  41. Krishnan AV, Starhis P, Permuth SF, Tokes L, Feldman D (1993) Bisphenol A: an estrogenic substance is released from polycarbonate flasks during autoclaving. Endocrinology 132:2279–2286CrossRefGoogle Scholar
  42. Kulk G, Poll WHVD, Visser RJW, Buma AGJ (2011) Distinct differences in photoacclimation potential between prokaryotic and eukaryotic oceanic phytoplankton. J Exp Mar Biol Ecol 398:63–72CrossRefGoogle Scholar
  43. Levy G, Lutz I, Kru¨ger A, Kloas W (2004) Bisphenol A induces feminization in Xenopus laevis tadpoles. Environ Res 94:102–111CrossRefGoogle Scholar
  44. Li R, Liu Y, Chen GZ, Tam NFY, Shin PKS, Cheng SG, Luan TG (2008) Physiological responses of the alga Cyclotella caspia to Bisphenol A exposure. Bot Mar 51(2):360–369Google Scholar
  45. Li R, Chen GZ, Tam NFY, Luan TG, Shin PKS (2009) Toxicity of bisphenol A and its bioaccumulation and removal by a marine microalga Stephanodiscus hantzschii. Ecotoxicol Environ Saf 72:321–328CrossRefGoogle Scholar
  46. Ma J, Lin F, Qin W, Wang P (2004) Differential response of four cyanobacterial and green algal species to triazophos, fentin acetate, and ethephon. Bull Environ Contam Toxicol 73:890–897CrossRefGoogle Scholar
  47. Maćczak A, Cyrkler M, Bukowska B, Michałowicz J (2017) Bisphenol A, bisphenol S, bisphenol F and bisphenol AF induce different oxidative stress and damage in human red blood cells (in vitro study). Toxicol In Vitro 41:143–149CrossRefGoogle Scholar
  48. Masojídek J, Grobbelaar JU, Pechar L, Koblízek M (2001) Photosystem electron transport rates and oxygen production in natural water blooms of freshwater cyanobacteria during a diel cycle. J Plankton Res 23:57–66CrossRefGoogle Scholar
  49. Massana R, Logares R (2013) Eukaryotic versus prokaryotic marine picoplankton ecology. Environ Microbiol 15:1254–1261CrossRefGoogle Scholar
  50. Melzer D, Rice N, Lewis C, Henley W, Galloway T (2010) Associacion of urinary bisphenol A concentration with heartdisease: evidence from NHANES 2003/06. PLoS 5:8673CrossRefGoogle Scholar
  51. Miao A, Wang W, Juneau P (2005) Comparison of Cd, Cu, and Zn toxic effects on four marine phytoplankton by pulse-amplitude-modulated fluorometry. Environ Toxicol Chem 24:2603–2611CrossRefGoogle Scholar
  52. Michałowicz J (2014) Bisphenol A—sources, toxicity and biotransformation. Environ Toxicol Pharmacol 37:738–758CrossRefGoogle Scholar
  53. Müller P, Li XP, Niyogi KK (2001) Non-photochemical quenching: a response to excess light energy. Plant Physiol 125:1558CrossRefGoogle Scholar
  54. Murthy KNC, Vanitha A, Rajesha J et al (2005) In vivo antioxidant activity of carotenoids from Dunaliella salina -a green microalga. Life Sci 76:1381–1390CrossRefGoogle Scholar
  55. Nusch EA (1980) Comparison of different methods for chlorophyll and pheopigment determination. Arch Hydrobiol Beiheft Ergebn Limnol 14:14–36Google Scholar
  56. Oehlmann J, Schulte-Oehlmann U, Tillmann M, Markert B (2000) Effects of endocrine disruptors on prosobranch snails (Mollusca:Gastropoda) in the laboratory. Part I: bisphenol A and octylphenol as xeno-estrogens. Ecotoxicol 9:383–397CrossRefGoogle Scholar
  57. Platt T, Gallegos CL, Harrison WG (1980) Photoinhibition of photosynthesis in natural assemblages of marine phytoplankton. J Mar Res 38:687–701Google Scholar
  58. Ran XF, Liu R, Xu S, Bai F, Xu J, Yang Y, Shi J, Wu Z (2015) Assessment of growth rate, chlorophyll a fluorescence, lipid peroxidation and antioxidant enzyme activity in Aphanizomenon flosaquae, Pediastrum simplex and Synedra acus exposed to cadmium. Ecotoxicology 24:46–177CrossRefGoogle Scholar
  59. Roháček K, Barták M (1999) Technique of the modulated chlorophyll fluorescence: basic concepts, useful parameters, and some applications. Photosynthetica 37:339–363CrossRefGoogle Scholar
  60. Sarchizian I, Ardelean II (2010) Axenic culture of a diazotrophic filamentous cyanobacterium isolated from mesothermal sulphurous springs (Obanul mare—Mangalia). Rom J Biol Plant Biol 55:47–53Google Scholar
  61. Schreiber U, Bilger W, Neubauer C (1995a) Chlorophyll fluorescence as a nonintrusive indicator for rapid assessment of in vivo photosynthesis. Springer, Berlin Heidelberg 100:49–70Google Scholar
  62. Schreiber U, Hormann H, Neubauer C, Klughammer C (1995b) Assessment of photosystem II photochemical quantum yield by chlorophyll fluorescence quenching analysis. Aust J Plant Physiol 22:209–220CrossRefGoogle Scholar
  63. Sonnenschein C, Soto AM (1998) An updated review of environmental estrogen and androgen mimics and antagonists. Steroid Biochem Mol Biol 65:143–150CrossRefGoogle Scholar
  64. Staples CA, Dorn PB, Klecka GM et al (1998) A review of the environmental fate, effects, and exposures of bisphenol A. Chemosphere 36:2149–2173CrossRefGoogle Scholar
  65. Stephenson RR (1982) Aquatic toxicology of cypermethrin. I. Acute toxicity to some freshwater fish and invertebrates in laboratory tests. Aquat Toxicol 2:175–185CrossRefGoogle Scholar
  66. Tao S, Zhang Y, Yuan C, Gao J, Wu F (2016) Oxidative stress and immunotoxic effects of bisphenol A on the larvae of rare minnow Gobiocypris rarus. Ecotoxicol Environ Saf 124:377–385CrossRefGoogle Scholar
  67. Tišler T, Krel A, Gerželj U, Erjavec B, Dolenc MS (2016) Hazard identification and risk characterization of bisphenols A, F, and AF to aquatic organisms. Environ Pollut 212:472–479CrossRefGoogle Scholar
  68. Uchiyama M, Mihara M (1978) Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Chem 86:271–278Google Scholar
  69. Vandenberg LN, Hauser R, Marcus M, Olea N, Welshons WV (2007) Human exposure to bisphenol A (BPA). Reprod Toxicol 24:139–177CrossRefGoogle Scholar
  70. Wakabayashi K, Böger P (1999) General physiological characteristics and mode of action of peroxidizing herbicides. In: Böger P, Wakabayashi K (eds) Peroxidizing herbicides. Springer-Verlag, Berlin, Germany, pp 163–190Google Scholar
  71. Wang ZC, Li DH, Li GW, Liu YD (2010) Mechanism of photosynthetic response in Microcystis aeruginosa PCC7806 to low inorganic phosphorus. Harmful Algae 9:613–619CrossRefGoogle Scholar
  72. Wang Q, Wang L, Han R, Yang L, Zhou Q (2015) Effects of bisphenol A on antioxidant system in soybean seedling roots. Environ Toxicol Chem 34:1127–1133CrossRefGoogle Scholar
  73. Weissman L, Garty J, Hochman A (2005) Characterization of enzymatic antioxidants in the lichen Ramalina lacera and their response to rehydration. Appl Environ Microbiol 71:6508–6514CrossRefGoogle Scholar
  74. Wu S, Shih MJ, Ho YC (2007) Toxicological stress response and cadmium distribution in hybrid tilapia (Oreochromis sp.) uponcadmium exposure. Toxicol Phamarcol 145:218–226Google Scholar
  75. Wu Z, Song L, Li R (2008) Different tolerances and responses to low temperature and darkness between water bloom forming cyanobacterium Microcystis and a green alga Scenedesmus. Hydrobiologia 596:47–55CrossRefGoogle Scholar
  76. Wu Z, Shi JQ, Yang SQ (2013) The effect of pyrogallic acid on growth, oxidative stress, and gene expression in Cylindrospermopsis raciborskii (Cyanobacteria). Ecotoxicology 22:271–278CrossRefGoogle Scholar
  77. Yang C, Yu XJ, Wang XZ, Kong HN, Li YH (2014) Effects of bisphenol A on the growth and physiology of Microcystis aeruginosa. Saf Environ Eng 21:21–25 (in Chinese) Google Scholar
  78. Zhang W, Bang X, Wen FS, Shuai A, Kuang FL, Mei JG, Xin HC (2014) Acute and chronic toxic effects of bisphenol a on Chlorella pyrenoidosa and Scenedesmus obliquus. Environ Toxicol 29:714–722CrossRefGoogle Scholar
  79. Ziv-Gal A, Zelieann C, Wang W, Flaws J (2013) Bisphenol A inhibits cultured mouse ovarian follicle growth partially viathe aryl hydrocarbon receptor signaling pathway. Reprod Toxicol 42:58–67CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Rong Xiang
    • 1
  • Junqiong Shi
    • 1
  • Yi Yu
    • 1
  • Hongbo Zhang
    • 1
  • Congcong Dong
    • 1
  • Yanjun Yang
    • 1
  • Zhongxing Wu
    • 1
  1. 1.Key Laboratory of Eco-environments in Three Gorges Reservoir Region (Ministry of Education), Chongqing Key Laboratory of Plant Ecology and Resources Research in Three Gorges Reservoir Region, School of Life ScienceSouthwest UniversityChongqingPeople’s Republic of China

Personalised recommendations