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1 2016 1, 23(1): Journal of Fishery Sciences of China 研究论文 DOI: /SP.J 鮨 1, 孔祥迪 2 1,, 刘莉 2, 李炎璐 2 1,, 于欢欢 2 2, 陈超 1., ; 2., 摘要 : (PAHs), 3 (Phe) 4 (Py) 5 (Bap) α- (ANF) 鮨 (Centropristis striata) (EROD) ( ), : (1)Bap Py EROD Phe, 3 PAHs 鮨 EROD Bap Py Phe,, % % %; ANF, EROD, % % % (2), 3 PAHs Bap Py Phe, 3, ; ANF, Bap Py, Phe ANF (P>0.05), 3 PAHs 鮨, Phe, Bap Py AHR CYP1 A : 鮨 ; ; ; ; 中图分类号 : X171 文献标志码 : A 文章编号 : (2016) (PAHs),,, [1] PAHs,,, [2 3],, PAHs,, [4 5] 鮨 (Centropristis striata) (Perciformes) (Epinephelinae),,, [6], 4~33, 5~36,,, [7], 鮨 [8 9] [10] 收稿日期 : ; 修订日期 : 基金项目 : (2012DFA30360); (2011BAD13B01). 作者简介 : (1988 ),,,. xiangdikong@163.com (1988 ),. 通信作者 :,. Tel: (0532) ; ysfrichenchao@126.com

2 [11] [12], PAHs PAHs 鮨, 鮨 PAHs, PAHs :, 2~3, (Phe),,, ;, 4~7, (Py) (Bap), AHR, PAH, P4501A (CYP1 A) [13] (Phe) (Py) (BaP) AHR CYPlA,, Bap Py Phe α - (ANF) CYPl A, CYPl A [14], Phe Py BaP ANF, 鮨 ; ANF CYPl A, Phe Py BaP 3 ANF 鮨,, PAHs CYPl A IU/kg (HCG), 1/2 48 h, 3.. 1,, 5 min, (21±0.5),,, 35(, 32), 5~10 min 1.2 Phe Py Bap ANF (ERF) Sigma,, ( 1), 2 Tab. 1 drug 表 1 PAHs 浓度梯度设计 The design of the PAHs concentration gradient PAHs /(µg L 1 ) concentration of PAHs Bap Py Phe ,, PAHs, PAHs, PAHs 100 µg/l ANF ; (EROD), ERF(21 µg/l in DMSO) PAHs PAHs ANF 1000 ml, 100, 3, 12 h 50 %, 4 h, 24 h 48 h 1.3 EROD [15], EROD 10~15 CYPlA 7- - 噁 (ERF) 噁,, 噁 CYPl A,, (NIS-Elements BR 4.10) (area meanintensity), EROD, EROD

1 : 3 鮨 243 1.4 [16 17] 10~15,, ( ),,,,, 0 1 2 3 ( 1) (DI)=(n /n ) 100 1 鮨 A. ; B. ; C. ; D.. Fig. 1 Heart malformation classification and assignment of embryo development in Centropristis striata A.

3 1 : 3 鮨 [16 17] 10~15,, ( ),,,,, ( 1) (DI)=(n /n ) 鮨 A. ; B. ; C. ; D.. Fig. 1 Heart malformation classification and assignment of embryo development in Centropristis striata A. Normal; B. Mild deformity; C. Moderate deformity; D. Severe deformity. 1.5 SPSS 17.0 ANOVA Duncan, ± ( x ±SD),, (P>0.05), (P<0.05) PAHs 鮨 EROD (Phe) EROD (98.47%) (100%), EROD Phe EROD 2, EROD Phe,, Phe 100 µg /L, EROD (P<0.05), Phe EROD 280.5%; Phe ANF, EROD, (P>0.05), Phe EROD (Py) Py EROD, (P<0.05, 2), EROD %; ANF, EROD, (P<0.05) Py EROD (Bap) Bap EROD Phe Py, 2.0 µg/l, Bap EROD (P<0.05), EROD %; ANF, EROD, (P<0.05), EROD 189.2%( 2) 鮨噁 PAHs 鮨 PAHs PAHs 鮨,

4 PAHs ANF 鮨 (EROD) a (P<0.05); b (P<0.05). Fig. 2 Three PAHs single exposure and the PAH+ANF combined exposure effect on embryonic ethoxyresorufin-o-deethylase (EROD) activity in Centropristis striata a denotes significant difference between exposed group and control group(p<0.05); b denotes significant differences between single exposure group and the combined exposure group (P<0.05). 3 PAH :, ; Phe, Bap Py,, ;,,,,,,,, PAHs ANF PAHs, Bap Py,, PAHs, PAHs Phe Py Bap 3 鮨 h 48 h (P>0.05), 鮨 Phe, 鮨,, 50 µg/l, 22.33±2.52, (0.00±0.00) (P<0.05); (88.25±0.49)%, [(91.82±2.64)%] (P>0.05) 500 g/l, (70.33± 0.58)% (72.56±2.21)%,,, 48 h (32.57±4.98)%; Phe ANF 鮨 Phe, 48 h (85.67±4.04)% (71.49± 0.07)% (26.62±2.71)%, (P<0.05) Py,, 10 µg/l, 32.33±2.52 (53.93±1.26)%,

5 1 : 3 鮨 245 (P<0.05), 500 μg/l, 80.00±0.00 (31.43±0.99)%,, 24 h, (100.00±0.00)%; Py ANF Py, 鮨, 48 h 73.58± 6.68 (78.09±2.53)% (24.28±4.00)% 3 鮨噁 ( Bap ) 1. ; 2. ; a d: μg/l Bap ; a* b*: μg/l Bap ANF. Fig. 3 Fluorescence images of embryosin resorufin in Centropristis striata (in the case of Bap) 1. Blank control group; 2. Acton group; a d. Groups under single exposure of 2, 4, 8, 20 µg/l Bap; a* b*. Groups under combined exposure of 2, 4, 8, 20 µg/l Bap and 100 µg/lanf ANF. PAHs Tab. 2 /(µg L 1 ) concentration 表 2 PAHs 单一暴露对条纹锯鮨鱼的毒性影响 Toxic effect of PAHs single exposure on Centropristis striata /% hatching rate /% mortality abnormality index 24 h 48 h acton 1 ml L ±1.55 a 4.40±1.91 a 4.56±1.28 a 8.64±0.87 a contrast ±2.64 a 0.00±0.00 a 2.55±1.47 a 2.23±0.40 a ±0.49 a 22.33±2.52 b 10.65±2.53 b 17.86±0.33 bc Phe ±1.17 ab 42.33±2.52 c 21.34±4.00 c 22.38±6.26 c ±5.28 bc 68.33±3.51 d 21.04±2.14 c 22.75±3.00 c ±2.21 c 70.33±0.58 d 22.72±1.55 c 32.57±4.98 d ±1.26 b 32.33±2.52 b 52.81±0.83 b 43.85±3.18 b Py ±0.92 c 46.05±6.76 c 87.54±0.56 c 69.67±4.04 c ±1.87 d 60.33±4.51 d 91.63±3.10 c ±0.00 d ±0.99 cd 80.00±0.00 e ±0.00 d ±0.71 b 8.20±4.11 b 17.65±1.43 b 26.93±0.85 b Bap ±3.30 c ±1.83 c 23.94±0.28 b 49.48±7.32 c ±4.45 d 51.34±1.16 d 55.18±4.54 c 67.80±3.90 d ±9.63 e 48.00±0.00 d 84.93±3.20 d 94.03±6.27 e : T,, (P<0.05). Note: Independence T test between acetone and control groups. Multiple comparison among control and Phe, Py, Bay treatments. Different letters denote significant difference (P<0.05).

6 PAHs+100µg/L ANF Tab. 3 /(µg L 1 ) concentration 表 3 PAHs 与 ANF 联合暴露对条纹锯鮨鱼的毒性影响 Toxic effect of combined exposure of PAHs and ANF on Centropristis striata /% hatching rate /% mortality abnormality index 24 h 48 h acton 1 ml/l 90.28±2.74 a 5.71±0.27 a 6.23±0.92 a 7.35±0.29 a contrast ±1.05 a 0.00±0.00 a 2.06±0.42 a 2.57±0.38 a ±3.48 b 51.46±1.70 b 14.67±4.51 b 12.70±2.08 b Phe ±0.35 c 56.73±3.34 b 22.33±1.53 c 18.59±0.32 c ±1.21 c 81.33±2.31 c 26.00±1.00 c 23.77±0.70 d ±0.07 c 85.67±4.04 c 32.33±1.53 d 26.62±2.71 d ±4.20 b 45.67±1.15 b 6.55±0.45 b 7.52±1.06 b Py ±5.05 b 60.82±5.84 c 12.60±0.53 c 15.04±2.18 c ±3.27 b 75.02±5.00 d 15.76±0.57 c 18.24±0.86 c ±2.53 b 73.58±6.68 d 19.87±2.99 d 24.28±4.00 d ±1.68 b 26.56±0.51 b 9.03±1.57 b 9.66±1.35 b Bap ±0.59 b 40.29±0.70 c 9.69±0.93 b 12.69±1.44 bc ±0.51 b 60.03±0.05 d 14.11±0.12 c 16.88±1.63 cd ±0.32 c 67.00±8.54 d 17.95±0.17 d 22.21±4.21 d : T,, (P<0.05). Note: Independence T test between acetone and control groups. Multiple comparison among control and Phe, Py, Bay treatments. Different letters denote significant difference (P<0.05). Bap,, ; 2 µg/l, 8.20±4.11 (73.47±0.71)%, (P< 0.05), 20 µg/l, (48.00±2.00) (28.60±9.63)%,, 24 h 48 h (84.93± 3.20)% (94.03±6.27)%; Bap ANF Bap, 鮨, 24 h 48 h 67.00±8.54 (64.64±0.32)% (17.95±0.17)% (22.21±4.21)% (Phe), Phe PAHs, AHR CYPI A, [18 19], Phe,, 鮨 EROD,,, 500 μg/l EROD , 50 µg/l 62.82% Phe,,, 17.80%, ( ) 48 h % 82.36% Phe ANF, EROD,,,, 500 µg/l, ANF %, Phe CYPI A, 鮨 Hawkins [20] 3.2 (Py), Py 鮨 EROD,, 24 h, 100 µg/l, 31.43±0.99 (80.00±

7 1 : 3 鮨 )% (100±0.00)%; Py ANF, EROD,, Py, 50%, 24 h 80%, CYPl A ANF, Py, ANF, Py,, Py 鮨 AHR CYPl A, CYPl A ANF, Py CYPl A,, 鮨 ; Py,, Py AHR, Py,,,, Py 鮨, Py [21] Incardona [22], Py AHR CYPlA, Wassenberg [16], Py ANF,,, Py,, :, AHR CYPl A, 3.3 (Bap) BaP, CYPl A,, CYPl A BaP, Bap 鮨 EROD, ; Bap ANF, EROD,, Bap, 40%; 48 h 70%,, CYPl A, CYPl A, ; CYPl A Bap : CYPl A,, BaP, BaP AHR, AHR Wills [17] Bap Killifish, FL CYPl A, Bap, Bap FL, Matson [23] CYPl A, CYPl s, BaP,, BaP ANF, CYPI A BaP, BaP AHR, : 3 鮨, EROD, 3 PAHs Bap Py Phe EROD (CYPI A), ; ANF, Bap Py, Phe, ANF PAHs,,,, PAHs, CYPI A

8 248 23, 参考文献 : [1] Wang X H, Zheng J S. POPs pollution in marine environment and its analysis and monitoring technology[m]. Beijing: China Ocean Press, 2011: [,. POPs [M]. :, 2011: ] [2] Peterson R E, Theobald H M, Kimmel G L Developmental reproductive toxicity of dioxins and related compounds: Cross-spe-cies comparisons[j]. Crit Rev Toxicol, 1993, 23: [3] Walker M k, Peterson R E. Aquatic toxicity of dioxins and related chemicals[m]//schecter A. Dioxins and Health. New York: Plenum, 1944: [4] White P A, Robitaille S, Rasmussen J B. Heritable reproductive effects of benzo[a]pyrene on the fathead minnow(pimephales promelas)[j]. Environ Toxicol Chem, 1999, 18: [5] Billiard S M, Hahn M E, Franks D G, et a1. Binding of polycyclic aromatic hydrocarbons(pahs) to teleost aryl hydrocarbon receptors[j]. Comp Biochem Physiol B, 2002, 133: [6] Lei J L, Lu J W. The breed predominance and culture prospects of Centropristis striata Linnaeus[J]. Marine Fisheries Research, 2007, 28(5): [,. [J]., 2007, 28(5): ] [7] Chu Y W. Seedling and cultivation technology of Centropristis striata[j]. Shandong Fishery, 2004, 21(1): 12. [. [J]., 2004, 21(1): 12.] [8] Qiu J H, Lin X. Analysis of nutrient and evaluation of nutritional value for Centropristis striata[j]. Journal of Hydroecology, 2009, 2(6): [,. [J]., 2009, 2(6): ] [9] Dang R, Zhu J Q, Qiu X Z. Analysis of flesh content and nutrient components in the muscle of Centropristis striata[j]. Journal of Marine Sciences, 2010, 28(2): [,,. [J]., 2010, 28(2): ] [10] Li H Y, Zhu J Q, Chen F, et al. The morphology of the digestive tract of Centropristis striata[j]. Journal of Biology, 2011, 28(4): [,,,. [J]., 2011, 28(4): ] [11] Jia R J, Wang L, Zhao C M. Preliminary studies on embryonic development and morphology of the yolk-sac larvae of Centropristis striata Linnaeus[J]. Progress in Fishery Sciences, 2012, 33(4): [,,,. [J]., 2012, 33(4): ] [12] Zhou M T. Experiment on sea cage culture of induced Centropristis striata[j]. Modern Fisheries Information, 2011, (8): [. [J]., 2011, (8): ] [13] Incardona J P, Day H L, Collier T K, et a1. Developmental toxicity of 4-ring polycyclic aromatic hydrocarbons in zebrafish is differentially dependent on AH receptor isoforms and hepatic cytochrome P4501A metabolism[j]. Toxicol Appl Pharmaeol, 2006, 217(3): [14] Mu J L, Wang X H, Jin F, et al. Effects of phenanthrene, pyrene and benzo(a) pyrene individual exposure and co-exposure with α-naphthoflavone on the embryonic development of marine medaka(oryzias melastigma)[j]. ACTA Oceanologica Sinica, 2012, 34(6): [,,,. (a) (ANF) (Oryzias melastigma) [J]., 2012, 34(6): ] [15] Cong W, Wang X H. Response of EROD activity of the Pagrus major during embryonic development exposed to benzo(a)pyrene[j]. Journal of Xiamen University: Natural Science, 2007, 46(1): [,. (a) 鯛 EROD [J]. :, 2007, 46(1): ] [16] Wassenberg D M, Di Giulio R T. Synergistic embryotoxicity of polycyclic aromatic hydrocarbon aryl hydrocarbon receptor agonists with cytochrome P4501A inhibitors in Fundulus heteroclitus[j]. Environ Health Perspect, 2004, 112: [17] Wills L P, Zhu S, Willett K L, et a1. Effect of CYP la inhibition on the biotransformation of benzo[a]pyrene in two populations of Fundulus heteroclitus with different exposure histories[j]. Aquat Toxicol, 2009, 92(3): , [18] Incardona J P, Collier T K, Scholz N L. Defects in cardiac function precede morphological abnormalities in fish embryos exposed to polycyclic aromatic hydrocarbons[j]. Toxicol Appl Pharmacol, 2004, 1 96: 19l 205. [19] Van Wezel A P, Opperhuizen A. Narcosis due to environmental pollutants in aquatic organisms: Residue-based toxicity, mechanisms, and membrane bur-dens[j]. Crit Rev Toxi-

9 1 : 3 鮨 249 col, 1995, 25(3): [20] Hawkins S A, Billiard S M, Tabash S P, et a1. Altering cytochrome P4501A activity affects polycyclic aromatic hydrocarbon metabolibm and toxicity in rainbow trout (Oncorhynchus mykiss)[j]. Environ Toxieol Chem, 2002, 21: [21] Teraoka H, Dong W, Tsujimoto Y, et a1. Induction of cytochrome P450-1A is required for circulation failure and edema by 2, 3, 7, 8-tetrachlorodibenzo P- dioxin in zebrafish[j]. Biochem Biophysical Res Commun, 2003, 304(2): [22] Incardona J P, Day H L, Collier T K, et a1. Developmental toxicity of 4-ring polycyclic aromatic hydrocarbons in zebrafish is differentially dependent on AH receptor isoforms and hepatic cytochrome P4501A metabolism[j]. Toxicol Appl Pharmacol, 2006, 217(3): [23] Matson C W, Timme-Laragy A R, Di Giulio R T. Fhloranthene, but not benzo[a]pyrene, interacts with hypoxia resulting in pericardial effusion and lordosis in developing zebrafish[j]. Chemosphere, 2008, 74(1): Toxic effects of three polycyclic aromatic hydrocarbons on early embryonic and larval development of Centropristis striata KONG Xiangdi 1, 2, LIU Li 1, 2, LI Yanlu 2, YU Huanhuan 1, 2, CHEN Chao 2 1. College of Fisheries and Science, Shanghai Ocean University, Shanghai , China; 2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao , China Abstract: We conducted this study to determine the toxicity mechanism of polycyclic aromatic hydrocarbons (PAHs) on the early development of the marine fish Centropristis striata. We analyzed individual exposure to phenanthrene (Phe), pyrene (Py), and benzo(a)pyrene (BaP) and co-exposure of each with α-naphthoflavone (ANF) to assess their toxic effects on early embryonic and larval development of C. striata. The results showed a clear dose-response relationship between Bap and Py concentration and ethoxyresorufin-o-deethylase (EROD) activity compared with that of Phe. The effects of the PAHs on EROD activity were in the order Bap (857.52% of control) > Py (514.21%) > Phe (280.50%). Maximum EROD activity occurred at the highest concentration compared with that of the control group. EROD activity was inhibited (189.27%, %, and % of the control) when ANF was added to Bap, Py, and Phe, respectively. Toxicity of the PAHs to hatching rate, deformity index, and early embryo and larval mortality was in the order of Bap > Py > Phe. PAH concentration and the percentage of fertilized eggs and hatching rate were negatively correlated but PAH concentration was positively correlated with larval mortality and the deformity index. Bap and Py showed reduced toxicity when combined with ANF. These results show that PAHs with different ring structures have obvious toxic effects, although there were differences in the mechanisms among them. Key words: Centropristis striata; PAHs; embryo; larvae; toxic effect Corresponding author: CHEN Chao. Tel: ; ysfrichenchao@126.com

Microsoft Word - 22 栗志民.doc

Microsoft Word - 22 栗志民.doc 44 6 Vol.44, No.6 2013 11 OCEANOLOGIA ET LIMNOLOGIA SINICA Nov., 2013 (Argopecten irradians concentricus) * 1 1 2 2 (1. 524025; 2. 524022) 2011 10 2012 4 (Argopecten irradians concentricus),,,,,,,, (P