Ecotoxicology

, Volume 23, Issue 2, pp 175–191 | Cite as

Toxicological effects of the sunscreen UV filter, benzophenone-2, on planulae and in vitro cells of the coral, Stylophora pistillata

  • C. A. Downs
  • Esti Kramarsky-Winter
  • John E. Fauth
  • Roee Segal
  • Omri Bronstein
  • Rina Jeger
  • Yona Lichtenfeld
  • Cheryl M. Woodley
  • Paul Pennington
  • Ariel Kushmaro
  • Yossi Loya
Article

Abstract

Benzophenone-2 (BP-2) is an additive to personal-care products and commercial solutions that protects against the damaging effects of ultraviolet light. BP-2 is an “emerging contaminant of concern” that is often released as a pollutant through municipal and boat/ship wastewater discharges and landfill leachates, as well as through residential septic fields and unmanaged cesspits. Although BP-2 may be a contaminant on coral reefs, its environmental toxicity to reefs is unknown. This poses a potential management issue, since BP-2 is a known endocrine disruptor as well as a weak genotoxicant. We examined the effects of BP-2 on the larval form (planula) of the coral, Stylophora pistillata, as well as its toxicity to in vitro coral cells. BP-2 is a photo-toxicant; adverse effects are exacerbated in the light versus in darkness. Whether in darkness or light, BP-2 induced coral planulae to transform from a motile planktonic state to a deformed, sessile condition. Planulae exhibited an increasing rate of coral bleaching in response to increasing concentrations of BP-2. BP-2 is a genotoxicant to corals, exhibiting a strong positive relationship between DNA-AP lesions and increasing BP-2 concentrations. BP-2 exposure in the light induced extensive necrosis in both the epidermis and gastrodermis. In contrast, BP-2 exposure in darkness induced autophagy and autophagic cell death. The LC50 of BP-2 in the light for an 8 and 24 h exposure was 120 and 165 parts per billion (ppb), respectively. The LC50s for BP-2 in darkness for the same time points were 144 and 548 ppb. Deformity EC20 levels (24 h) were 246 parts per trillion in the light and 9.6 ppb in darkness.

Keywords

Coral Benzophenone-2 Cell toxicity Coral planula Sunscreen UV filters 

Supplementary material

10646_2013_1161_MOESM1_ESM.tif (654 kb)
Proportion of planula alive as a function of benzophenone-2 concentration. Symbol size is proportional to the number of replicates having a given value. Regression line (solid) and 95 % confidence intervals (dashed lines) are shown for each statistically significant fit. aStylophora pistillata exposed to benzophenone-2 for 8 h in the light. bS. pistillata exposed to benzophenone-2 for 8 h in the dark. cS. pistillata exposed to benzophenone-2 for planulae exposed for 8 h in the light, then 16 h of darkness. dS. pistillata exposed to benzophenone-2 for 24 h in the dark
10646_2013_1161_MOESM2_ESM.tif (781 kb)
Proportion of non-deformed planulae as a function of benzophenone-2 concentration. Symbol size is proportional to the number of replicates having a given value. Regression line (solid) and 95 % confidence intervals (dashedlines) are shown for each statistically significant fit. aStylophora pistillata exposed to benzophenone-2 for 8 h in the light. bS. pistillata exposed to benzophenone-2 for 8 h in the dark. cS. pistillata exposed to benzophenone-2 for planulae exposed for 8 h in the light, then 16 h of darkness. dS. pistillata exposed to benzophenone-2 for 24 h in the dark
10646_2013_1161_MOESM3_ESM.tif (1.4 mb)
PROBIT analyses of dose–response curves of cells of Stylophora pistillata exposed to benzophenone-2. PROBITs fit using a Gompertz–Weibull exponential distribution were used to calculate LC50s. aS. pistillata exposed to benzophenone-2 for 8 h in the light. bS. pistillata exposed to benzophenone-2 for 8 h in the dark
10646_2013_1161_MOESM4_ESM.tif (1.2 mb)
PROBIT analyses of dose–response curves of number of deformed planulae of Stylophora pistillata exposed to benzophenone-2. PROBITs fit using a Gompertz-Weibull exponential distribution were used to calculate EC50s. aS. pistillata exposed to benzophenone-2 for 8 h in the light. bS. pistillata exposed to benzophenone-2 for planulae exposed for 8 h in the light, then 16 h of darkness. cS. pistillata exposed to benzophenone-2 for 8 h in the dark. dS. pistillata exposed to benzophenone-2 for 24 h in the dark
10646_2013_1161_MOESM5_ESM.tif (898 kb)
PROBIT analyses of dose-lethality curves of planulae of Stylophora pistillata exposed to benzophenone-2. PROBITs fit using a Gompertz–Weibull exponential distribution were used to calculate LC50s. aS. pistillata exposed to benzophenone-2 for 8 h in the light. bS. pistillata exposed to benzophenone-2 for 24 h in the light. cS. pistillata exposed to benzophenone-2 for 8 h in the dark

References

  1. Abelson A, Ronen O, Gaines S (2005) Coral recruitment to the reefs of Eilat, Red Sea: temporal and spatial variation, and possible effects of anthropogenic disturbances. Mar Pollut Bull 50:576–582Google Scholar
  2. Agati G, Mazzinghi P, Fusi F, Ambrosini I (1995) The F685/F730 chlorophyll fluorescence ratio as a tool in plant physiology: response to physiological and environmental factors. J Plant Physiol 14:228–238Google Scholar
  3. Agresti A (2002) Categorical data analysis, 2nd edn. Wiley, New YorkGoogle Scholar
  4. Anderson SL, Wild GC (1994) Linking genotoxic responses and reproductive success in ecotoxicology. Environ Health Perspect 102:9–12Google Scholar
  5. Aquera A, Martinez Bueno MJ, Fernandez-Alba AR (2013) New trends in the analytical determination of emerging contaminants and their transformation products in environmental waters. Environ Sci Pollut Res Int 20:3496–3515Google Scholar
  6. Bishop CD, Huggett MJ, Heyland A, Hodion J, Brandhorst BP (2006) Interspecific variation in metamorphic competence in marine invertebrates: the significance for comparative investigations into the timing of metamorphosis. Integr Comp Biol 46:662–682Google Scholar
  7. Blaauboe BJ (2008) The contribution of in vitro toxicity data in hazard and risk assessment: current limitations and future perspective. Toxicol Lett 180:81–84Google Scholar
  8. Blitz JB, Norton SA (2008) Possible environmental effects of sunscreen run-off. J Am Acad Dermatol 59:898. doi:10.1016/j.jaad.2008.06.013 Google Scholar
  9. Brooks AC, Gaskell PN, Maltby LL (2009) Importance of prey and predator feeding behaviors for trophic transfer and secondary poisoning. Environ Sci Technol 43:7916–7923Google Scholar
  10. Burke RD (1983) The induction of metamorphosis of marine invertebrate larvae: stimulus and response. Can J Zool 61:1701–1719Google Scholar
  11. Carson FL (1997) Histotechnology: a self-instructional text, 2nd edn. American Society of Clinical Pathologists, ChicagoGoogle Scholar
  12. CIR (Cosmetic Ingredient Review) (2005) Annual review of cosmetic ingredient safety assessments—2003/2003. Int J Toxicol 24:1–102Google Scholar
  13. Cosnefroy A, Brion F, Maillot-Marechal E et al (2012) Selective activation of zebrafish estrogen receptor subtypes by chemicals by using stable reporter gene assay developed in a zebrafish liver cell line. Toxicol Sci 125:439–449Google Scholar
  14. Crawley MJ (1993) GLIM for ecologists. Blackwell, LondonGoogle Scholar
  15. Cuquerella MC, Lhiaubet-Vallet V, Cadet J, Miranda MA (2012) Benzophenone photosensitized DNA damage. Acc Chem Res 45:1558–1570Google Scholar
  16. Danovaro R, Bongiorni L, Corinaldesi C et al (2008) Sunscreens cause coral bleaching by promoting viral infections. Environ Health Persp 116:441–447Google Scholar
  17. Daughton CG (2002) Environmental stewardship and drugs as pollutants. Lancet 360:1035–1036Google Scholar
  18. Depledge MH (1998) The ecotoxicological significance of genotoxicity in marine invertebrates. Mutat Res 399:109–122Google Scholar
  19. Downs CA, Fauth JE, Halas JC, Dustan P, Bemiss J, Woodley CM (2002) Oxidative stress and seasonal coral bleaching. Free Radical Biol Med 32:533–543Google Scholar
  20. Downs CA, Karamarsky-Kramer W, Martinez J et al (2009) Symbiophagy as a mechanism for coral bleaching. Autophagy 5:211–216Google Scholar
  21. Downs CA, Fauth JE, Downs VD, Ostrander GK (2010) In vitro cell-toxicity screening as an alternative animal model for coral toxicology: effects of heat stress, sulfide, rotenone, cyanide, and cuprous oxide on cell viability and mitochondrial function. Ecotoxicology 19:171–184Google Scholar
  22. Downs CA, Woodley CM, Fauth JE et al (2011) A survey of environmental pollutants and cellular-stress biomarkers of Porites astreoides at six sites in St. John. US Virgin Islands. Ecotoxicology 20:1914–1931Google Scholar
  23. Downs CA, Ostrander GK, Rougee L et al (2012) The use of cellular diagnostics for identifying sub-lethal stress in reef corals. Ecotoxicology 21:768–782Google Scholar
  24. Downs CA, McDougall KE, Woodley CM et al (2013) Heat stress and light stress induce different cellular pathologies in the symbiotic dinflagellate during coral bleaching. PLoS ONE 8(12):e77173Google Scholar
  25. Drablos F, Feyzi E, Aas PA et al (2004) Alkylation damage in DNA and RNA—repair mechanisms and medical significance. DNA Repair 3:1389–1407Google Scholar
  26. Draper NR, Smith H (1966) Applied regression analysis. Wiley, New YorkGoogle Scholar
  27. Dustan P (1977) Vitality of reef coral populations off Key Largo, Florida: recruitment and mortality. Environ Geol 2:51–58Google Scholar
  28. EC (2003) Technical Guidance Documents on Risk Assessment, Part II. EUR 20418 EN/2 Ispra, Italy: European Commission, Joint Research Centre, http://ihcp.jrc.ec.europa.eu/our_activities/public-health/risk_assessment_of_Biocides/doc/tgd/tgdpart2_2ed.pdf. Accessed 18 Nov 2013
  29. Edinger EN, Jompa J, Limmon GV, Widjatmoko W, Risk MJ (1998) Reef degradation and coral biodiversity in Indonesia: effects of land-based pollution, destructive fishing practices and changes over time. Mar Pollut Bull 36:617–630Google Scholar
  30. Eichenseher T (2006) The cloudy side of sunscreens. Environ Sci Technol 40:1377–1378Google Scholar
  31. Eskelinin EL, Reggiori F, Baba M, Kovacs AL, Seglen PO (2011) Seeing is believing: the impact of electron microscopy on autophagy research. Autophagy 7:935–956Google Scholar
  32. Fadlallah YH (1983) Sexual reproduction, development and larval biology in scleractinian corals: a review. Coral Reefs 2:129–150Google Scholar
  33. Finney DJ (1947) Probit analysis, a statistical treatment of the sigmoid response curve. Cambridge University Press, CambridgeGoogle Scholar
  34. Fortini P, Raspaglio G, Falchi M, Dogliotti E (1996) Analysis of DNA alkylation damage and repair in mammalian cells by the COMET assay. Mutagen 11:169–175Google Scholar
  35. Futch JC, Griffin DW, Lipp EK (2010) Human enteric viruses in groundwater indicate offshore transport of human sewage to coral reefs of the Upper Florida Keys. Environ Microbiol 12:964–974Google Scholar
  36. Gago-Ferrero P, Díaz-Cruz MS, Barceló D (2011) Occurrence of multiclass UV filters in treated sewage sludge from wastewater treatment plants. Chemosphere 84:1158–1165Google Scholar
  37. Gilbert E, Pirot F, Bertholle V, Roussel L, Falson F, Padois K (2013) Commonly used UV filter toxicity on biological functions: review of last decade studies. Int J Cosmetic Sci 35:208–219Google Scholar
  38. Gitelson AA, Buschmann C, Lichtenthaler HK (1999) The chlorophyll fluorescence ration F735/F700 as an accurate measure of the chlorophyll content in plants. Remote Sensing Environ 69:296–302Google Scholar
  39. Gleason DF, Hofmann DK (2011) Coral larvae: from gametes to recruits. J Exp Mar Biol Ecol 408:42–57Google Scholar
  40. Golbuu Y, Fabricius K, Victor S, Richmond R (2008) Gradients in coral reef communities exposed to muddy river discharges in Pohnpei, Micronesia. Estuar Coast Shelf S 76:14–20Google Scholar
  41. Gura T (2008) Toxicity testing moves from the legislature to the Petri dish–and back. Cell 134:557–559. doi:10.1016/j.cell.2008.08.011 Google Scholar
  42. Harii S, Nadaoka K, Yamamoto M, Iwao K (2007) Temporal changes in settlement, lipid content, and lipid composition of larvae of the spawning hermatypic coral Acropora tenuis. Mar Ecol-Prog Ser 346:86–89Google Scholar
  43. Harper CA, Petrie EM (2003) Plastics materials and processes: a concise encylopedia. Wiley, HobokenGoogle Scholar
  44. Hsieh MH, Grantham EC, Liu B, Macapagal R, Willingham E, Baskin LS (2007) In utero exposure to benzophenone-2 causes hypospadias through an estrogen receptor dependent mechanism. J Urology 178:1637–1642Google Scholar
  45. Hughes TP, Tanner JE (2000) Recruitment failure, life histories, and long-term decline of Caribbean corals. Ecology 81:2250–2263Google Scholar
  46. Jager T, Heugens EHW, Kooijman SALM (2006) Making sense of ecotoxicological test results: towards application of process-based models. Ecotoxicology 15:305–314Google Scholar
  47. Jarry H, Christoffel J, Rimoldi G, Koch L, Wuttke W (2004) Multi-organic endocrine disrupting activity of the UV screen benzophenone 2 (BP2) in ovariectomized adult rats after 5 days treatment. Toxicology 205:87–93Google Scholar
  48. Kerdivel G, Le Guevel R, Habauzit D, Brion F, Ait-Aissa S, Pakdel F (2013) Estrogenic potency of benzophenone UV filters in breast cancer cells: proliferative and transcriptional activity substantiated by docking analysis. PLoS ONE 8:e60567. doi:10.1371/journal.pone.0060567 Google Scholar
  49. Kerr JFR, Wullie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implication in tissue kinetics. Br J Cancer 26:239–257Google Scholar
  50. Kim Y, Ryu JC, Choi H-S, Lee K (2011) Effect of 2,2′,4,4′-tetrahydroxybenzopheonone (BP2) on steroidogenesis in testicular Leydig cells. Toxicology 288:18–26Google Scholar
  51. Klionsky DJ, Abdalalla FC, Abeliovich H et al (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8:445–544Google Scholar
  52. Knowland J, McKenzie EA, McHugh PJ, Cridland NA (1993) Sunlight-induced mutagenicity of a common sunscreen ingredient. FEBS Lett 324:309–313Google Scholar
  53. Koda T, Umezu T, Kamata R, Morohoshi K, Ohta T, Morita M (2005) Uterotrophic effects of benzophenone derivatives and a p-hydroxybenzoate used in ultraviolet screens. Environ Res 98:40–45Google Scholar
  54. Krysko DV, Vanden Berghe Y, Parthoens E, D’Herde K, Vandenabeele P (2008) Methods for distinguishing apoptotic from necrotic cells and measuring their clearance. Methods Enzymol 442:307–341Google Scholar
  55. Kuba K, Ide H, Wallace SS, Kow YK (1992) A novel, sensitive and specific assay for abasic sites, the most commonly produced DNA lesion. Biochemistry–US 31:3703–3708Google Scholar
  56. Kunisue T, Chen Z, Buck Louis GM et al (2012) Urinary concentrations of benzopheone-type UV filters in U.S. women and their association with endometriosis. Environ Sci Technol 46:4624–4632Google Scholar
  57. Kunz PY, Fent K (2009) Estrogenic activity of ternary UV filter mixtures in fish (Pimephales promelas) and analysis with nonlinear isobolograms. Toxicol Appl Pharm 234:77–88Google Scholar
  58. Kunz PY, Galicia HF, Fent K (2006) Comparison of in vitro and in vivo estrogenic activity of UV filters in fish. Toxicol Sci 90:349–361Google Scholar
  59. Kvitt H, Rosenfeld H, Zandbank K, Tchernov D (2011) Regulation of apoptotic pathways by Stylophora pistillata to survive thermal stress and bleaching. PLoS ONE 6:e28665Google Scholar
  60. Laskowski R (1995) Some good reasons to ban the use of NOEC, LOEC, and related concepts in Ecotoxicology. Oikos 73:140–144Google Scholar
  61. Lichtenthaler HK (1987) Chlorophyll and carotenoids, the pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382Google Scholar
  62. Miller MW, Weil E, Szmant AM (2000) Coral recruitment and juvenile mortality as structuring factors for reef benthic communities in Biscayne National Park, USA. Coral Reefs 19:115–123Google Scholar
  63. Molina-Molina J-M, Escande A, Pillon A et al (2008) Profiling of benzopheone derivatives using fish and human estrogen receptor-specific in vitro bioassays. Toxicol Appl Pharm 232:384–395Google Scholar
  64. Morohoshi K, Yamamoto H, Kamata R, Shiraishi F, Koda T, Morita M (2005) Estrogenic activity of 37 components of commercial sunscreen lotions evaluated by in vitro assays. Toxicol In Vitro 19:457–469Google Scholar
  65. Nashez LG, Schuster D, Laggner C et al (2010) The UV-filter benzophenone-1 inhibits 17 beta-hydrozysteroid dehydrogenase type 3: virtual screening as a strategy to identify potential endocrine disrupting chemicals. Biochem Pharmacol 79:1189–1199Google Scholar
  66. Nesa B, Baird AH, Harii S, Yakovleva I, Hidaka M (2012) Algal symbionts increase DNA damage in coral planulae exposed to sunlight. Zool Stud 51:12–17Google Scholar
  67. Newman MC (2013) Quantitative ecotoxicology. CRC Press, Boca RatonGoogle Scholar
  68. NTP (National Toxicology Program) (2006) NTP technical report on the toxicology and carcinogenesis of benzophenone in F344/N rats and B6C3F1 mice. NIH Publication #06-4469Google Scholar
  69. Omori M (2011) Degradation and restoration of coral reefs: experience in Okinawa, Japan. Mar Biol Res 7:3–12Google Scholar
  70. Paxton CW, Davy SK, Weis VM (2013) Stress and death of cnidarian host cells play a role in cnidarian bleaching. J Exp Biol 216:2813–2820Google Scholar
  71. Pitarch E, Portolés T, Marín JM et al (2010) Analytical strategy based on the use of liquid chromatography and gas chromatography with triple-quadrupole and time-of-flight MS analyzers for investigating organic contaminants in wastewater. Anal Bioanal Chem 397:2763–2776Google Scholar
  72. Platt KL, Aderhold S, Kulpe K, Fickler M (2008) Unexpected DNA damage caused by polycyclic aromatic hydrocarbons under standard laboratory conditions. Mut Res 650:96–103Google Scholar
  73. Popkin DJ, Prival MJ (1985) Effects of pH on weak and positive control mutagens in the AMES Salmonella plate assay. Mut Res 142:109–113Google Scholar
  74. Rees JG, Setiapermana D, Sharp VA, Weeks JM, Williams TM (1999) Evaluation of the impacts of land-based contaminants on the benthic faunas of Jakarta Bay, Indonesia. Oceano Acta 22:627–640Google Scholar
  75. Richardson SD (2006) Environmental mass spectrometry: emerging contaminants and current issues. Anal Chem 78:4021–4046Google Scholar
  76. Richardson SD (2007) Water analysis: emerging contaminants and current issues. Anal Chem 79:4295–4324Google Scholar
  77. Richmond R (1993) Coral reefs: present problems and future concerns resulting from anthropogenic disturbance. Amer Zool 33:524–536Google Scholar
  78. Richmond R (1997) Reproduction and recruitment in corals: critical links in the persistence of reefs. Life and death of coral reefs. Chapman and Hall, New York, pp 175–197Google Scholar
  79. Rodil R, Quintana JB, Concha-Grana E, Lopex-Mahia P, Muniatequi-Lorenzo S, Prada-Rodriguez D (2012) Emerging pollutants in sewage, surface and drinking water in Galicia (NW Spain). Chemosphere 86:1040–1049Google Scholar
  80. Samara P, Syntichaki N, Tavernarakis N (2008) Autophagy is required for necrotic cell death in Caenorhabditis elegans. Cell Death Differ 15:105–112Google Scholar
  81. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Long IslandGoogle Scholar
  82. Schlecht C, Klammer H, Wolfgang W, Jarry H (2006) A dose-response study on the estrogenic activity of benzophenone-2 on various endpoints in the serum, pituitary and uterus of female rats. Arch Toxicol 80:656–661Google Scholar
  83. Schlumpf M, Cotton B, Conscience M, Haller V, Steinmann B, Lichtensteiger W (2001) In vitro and in vivo estrogenicity of UV screens. Environ Health Persp 109:239–244Google Scholar
  84. Schlumpf M, Schmid P, Durrer S et al (2004) Endocrine activity and developmental toxicity of cosmetic UV filters—an update. Toxicology 205:113–122Google Scholar
  85. Schlumpf M, Durrer S, Fass O et al (2008) Developmental toxicity of UV filters and environmental exposure: a review. Int J Androl 31:144–151Google Scholar
  86. Schmutzler C, Bacinski A, Gotthardt I et al (2007) The ultraviolet filter benzophenone 2 interferes with the thyroid hormone axis in rats and is a potent in vitro inhibitor of human recombinant thyroid peroxidase. Endocrinology 148:2835–2844Google Scholar
  87. Scholze M, Boedeker W, Faust M, Backhaus T, Altenburger R, Grimme LH (2001) A general best-fit method for concentration-response curves and the estimation of low-effect concentrations. Environ Toxicol Chem 20:448–457Google Scholar
  88. Seidlova-Wuttke D, Jarry H, Wuttke W (2004) Pure estrogenic effect of benzophenone-2 (BP2) but not of bisphenol A (BPA) and dibutylphtalate (DBP) in uterus, vagina and bone. Toxicology 205:103–112Google Scholar
  89. Seidlova-Wuttke D, Jarry H, Christoffel J, Rimoldi G, Wuttke W (2005) Effects of bisphenol-A, dibutylphtalate, benzophenone-2, procymidone, and linurone on fat tissue, a variety of hormones and metabolic parameters: a 3 months comparison with effect of estradiol in ovariectomized rats. Toxicology 213:13–24Google Scholar
  90. Shaath NA, Shaath M (2005) Recent sunscreen market trends. In: Shaath NA (ed) Sunscreens, regulations and commercial development, 3rd edn. Taylor & Francis, Boca Raton, pp 929–940Google Scholar
  91. Smith TB, Nemeth RS, Blondeau J, Calnan JM, Kadison E, Herzlieb S (2008) Assessing coral reef health across onshore to offshore stress gradients in the US Virgin Islands. Mar Pollut Bull 56:1983–1991Google Scholar
  92. Song M, Kim Y-J, Park Y-K, Ryu J-C (2012) Changes in thyroid peroxidase activity in response to various chemicals. J Environ Monitor 14:2121–2127Google Scholar
  93. Spangenberg DB (1971) Thyroxine induced metamorphosis in Aurelia. J Exp Zoo 178:183–194Google Scholar
  94. Sung J-S, Demple B (2006) Roles of base excision repair subpathways in correcting oxidized abasic sites in DNA. FEBS J 273:1620–1629Google Scholar
  95. Suter GW II (1993) Ecological risk assessment. CRC Press, Boca RatonGoogle Scholar
  96. Suter, II GW, Mabrey JB (1994) Toxicological benchmarks for screening potential contaminants of concern for effects on aquatic biota: 1994 Revision. Oak Ridge National Laboratory, Oak Ridge, TN. ES/ER/TM-96/R1Google Scholar
  97. Suter GW II, Rosen AE, Linder E, Parkhurst DF (1987) End points for responses of fish to chronic toxic exposures. Environ Toxicol Chem 6:793–809Google Scholar
  98. Suzuki T, Kitamura S, Khota R, Sugihara K, Fujimoto N, Ohta S (2005) Toxicol Appl Pharm 203:9–17Google Scholar
  99. Taatjes DJ, Sobel BE, Budd RC (2008) Morphological and cytochemical determination of cell death by apoptosis. Histochem Cell Biol 129:33–43Google Scholar
  100. Tasdemir Em Galluzzi L, Majuri MN et al (2008) Methods for assessing autophagy and autophagic cell death. Methods Mol Biol 445:29–76Google Scholar
  101. Thienpont B, Tingaud-Sequeira A, Prats E, Barata C, Babin P, Raldua D (2011) Zebrafish eleutheroembryos provide a suitable vertebrate model for screening chemicals that impair thyroid hormone synthesis. Environ Sci Technol 45:7525–7532Google Scholar
  102. Tsujimoto Y, Shimizu S (2005) Another way to die: autophagic programmed cell death. Cell Death Differ 15:1528–1534Google Scholar
  103. Weisbrod CJ, Kunz PY, Zenker AK, Fent K (2007) Effects of the UV filter benzophenone-2 on reproduction in fish. Toxicol Appl Pharm 225:255–266Google Scholar
  104. Wennig R (2000) Threshold values in toxicology—useful or not? Forensic Sci Int 113:323–330Google Scholar
  105. West JM, Salm RV (2003) Resistance and resilience to coral bleaching: implications for coral reef conservation and management. Conserv Biol 17:956–967Google Scholar
  106. White MK, Cinti C (2004) A morphologic approach to detect apoptosis based on electron microscopy. Methods Mol Biol 285:105–111Google Scholar
  107. Williams DE, Miller MW, Kramer KL (2008) Recruitment failure in Florida Keys Acropora palmata, a threatened Caribbean coral. Coral Reefs 27:697–705Google Scholar
  108. Wilson DM, Barsky D (2001) The major human basic endonuclease: formation, consequences and repair of abasic lesions in DNA. Mut Res 485:283–307Google Scholar
  109. Yamasaki K, Takeyoski M, Yakabe Y, Sawaki M, Takatsuki M (2003) Comparison of the reporter gene assay for ER-alpha antagonists with the immature rat uterotrophic assay of 10 chemicals. Toxicol Lett 142:119–131Google Scholar
  110. Ye L, Su Z-J, Ge R-S (2011) Inhibitors of testosterone biosynthetic and metabolic activation enzymes. Molecules 16:9983–10001Google Scholar
  111. Yla-Antilla P, Vihinen H, Jokitalo E, Eskelinin EL (2009) Monitoring autophagy by electron microscopy in mammalian cells. Methods Enzymol 452:143–164Google Scholar
  112. Yu H (2002) Environmental carcinogenic polycyclic aromatic hydrocarbons: photochemistry and phototoxicity. J Environ Sci Heal, Part C 20:149–183Google Scholar
  113. Zar JH (1996) Biostatistical analysis, 3rd edn. Prentice Hall, Upper Saddle RiverGoogle Scholar
  114. Zeiger E, Anderson B, Haworth S, Lawlow T, Mortlemans K, Speck W (1987) Salmonella mutagenicity tests: 3. Results from the testing of 255 chemicals. Environ Mutagen 9:1–110Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • C. A. Downs
    • 1
  • Esti Kramarsky-Winter
    • 2
    • 3
  • John E. Fauth
    • 4
  • Roee Segal
    • 2
  • Omri Bronstein
    • 2
  • Rina Jeger
    • 5
  • Yona Lichtenfeld
    • 3
  • Cheryl M. Woodley
    • 6
    • 7
  • Paul Pennington
    • 7
  • Ariel Kushmaro
    • 3
    • 8
  • Yossi Loya
    • 2
  1. 1.Haereticus Environmental LaboratoryCliffordUSA
  2. 2.Department of Zoology, George S. Wise Faculty of Life SciencesTel Aviv UniversityTel AvivIsrael
  3. 3.Avram and Stella Goldstein-Goren Department of Biotechnology Engineering, Faculty of Engineering Sciences and National Institute For Biotechnology in the NegevBen-Gurion University of the NegevBeer ShevaIsrael
  4. 4.Department of BiologyUniversity of Central FloridaOrlandoUSA
  5. 5.Department of Life SciencesBen-Gurion University of the NegevBeer ShevaIsrael
  6. 6.Hollings Marine LaboratoryU.S. National Oceanic & Atmospheric AdministrationCharlestonUSA
  7. 7.Center for Coastal Environmental Health and Biomolecular ResearchU.S. National Oceanic & Atmospheric AdministrationCharlestonUSA
  8. 8.School of Materials Science & EngineeringNanyang Technological UniversitySingaporeSingapore

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