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1 43 6 Vol.43, No OCEANOLOGIA ET LIMNOLOGIA SINICA Nov., 2012 (Verasper moseri) * 1, 2 1, 2 1, 2 1, 2 ( ; ) (sgnrha) (HCG), : sgnrha HCG,, (T) (E 2 ), 96h, sgnrha, HCG,,,, S965 (Verasper moseri), (Pleuronectiformes) (Pleuronectoridei) (Pleuronectidae) (Verasper),,,,,,,,, (,, ), (Pleuronectes ferrugineus) (Scophthalmus maximus)(zohar et al, 2001), (Berlinsky et al, 1997) (Vermeirssen et al, 2000),, (GnRH),,, (Clearwater et al, 1995; Vermeirssen et al, 1998; Pankhurst, 1994), (Pleuronectes americanus) (Harmin et al, 1993) (Alveriño et al, 1993) (Larsson et al, 1997) (Paralichthys lethostigma)(berlinsky et al, 1996) (Paralichthys dentatus) *, CARS-50 ; 863, 2012AA10A413 ;, ;, ,,, xuyj@ysfri.ac.cn :, liuxz@ysfri.ac.cn : , :

ferrugineus) (Scophthalmus maximus)(zohar et al, 2001), (Berlinsky et al, 1997) (Vermeirssen et al, 2000),, (GnRH),,, (Clearwater et al, 1995; Vermeirssen et al, 1998; Pankhurst, 1994), (Pleuronectes

2 6 : (Verasper moseri) 1171 (Berlinsky et al, 1997) (Hippoglossus hippoglossus)(vermeirssen et al, 2000),, (Weil et al, 1983), (Anguilla marmorata)(, 2010) (Epinephelus akaara)(, 2005) (Paralichthys olivaceus)(, 2006), (HCG) (LHRH-a),,,,,,, , (Verasper moseri), 25 33cm, g , 29 31, ph , 6mg/L, 400% 600% 2, 1% 2%, : (3 ) 3 9 MS222 (150mg/L),, 30% 40%, 1 6min, 3 6ml 1.3 4m 3 3, 3, 3, 1.2 (HCG) (sgnrha)( ),, 1 15d 3d, (0h) ( 1), 1ml, 2cm, 180mg/L MS h, 表 1 实验设计用激素种类及注射剂量和亲鱼使用数量表 Tab.1 Hormones, dosages and number of experimental broodstocks used in this study ( ) (0.85%) 0.5ml/kg 3 sgnrha sgnrha 0.5ml/kg 3 HCG HCG 400IU/kg (0.5ml) h 96h 192h 0.5ml, 180mg/L MS-222,,,,,, 1.5ml,, ( ), 1.5 (1999) : 1 3 1,,, :

2, 6mg/L, 400% 600% 2, 1% 2%, : (3 ) 3 9 MS222 (150mg/L),, 30% 40%, 1 6min, 3 6ml 1.3 4m 3 3, 3, 3, 1.

3 ; 90% ; 90% ; 90% : 1 (,,, 30%) 2 (,, 50% 75%) 3 (,, 80%) 3,,, 6d, 1.6 (0h) 6h 12h 24h 48h 96h( 48h) 192h( ), (T) (E 2 ) 180mg/L MS-222 1ml,, 1.5ml, 2.0ml Eppendorf, r/min 10min,, 40 (E 2 ) (T) (RIA) (, 2007) 125 I E 2 T ( ) T 95% 108%, 1.9ng/dl, 7.4%, 9.8%, , , PROG , E 3 ( ) , E E 2 95% 104%, 2.1pg/ml, 7.7%, 8.9%, E , PROG , TESTO , , %, 30% 1.7 S.D., SPSS(16.0 ) (ANOVA), (P) 0.05, P 0.05, 2,, 3 6ml/, 1A 1B ( ) 1C D E F 2.1, 48h, HCG sgnrha, 96h, (P<0.05), 192h,, : HCG 3,, 85.0% 89.9%, 82.0% 84.6%, (P<0.05), min ( 1C, D, E, F) 48h, sgnrha 3,, 90.5% 98.2%, 84.7% 92.1%, (P<0.05), min( 1C, D, E, F) 6d,, 2 ( 1C), 1,, 2.2 6h, T E 2 (P<0.05), (P>0.05), HCG T 6h (P>0.05), 12h, 17.5ng/dl (12h) 49.31ng/dl (48h), 48h 96h 56.58ng/dl (P<0.05), 192h 37.33ng/dl sgnrha T 6h, 12h, 48h 78.13ng/dl, (P<0.05),, 96h T

1pg/ml, 7.7%, 8.9%, E 3 9.0 10 4, PROG 1.0 10 4, TESTO 1.0 10 4, 7.0 10 3, 1.0 10 5 3%, 30% 1.7 S.D., SPSS(16.0 ) (ANOVA), (P) 0.05, P 0.05, 2,, 3 6ml/, 1A 1B ( ) 1C D E F 2.

4 6 : (Verasper moseri) Fig.1 Changes of serum sex steroids levels and sperm quality after exogenous hormonal induction A., B., C., D., E., F. 48h (P>0.05), 192h, (P<0.05) 192h (sgnrha HCG, P<0.05), sgnrha T HCG T (P>0.05) E 2, HCG E 2 6h, 24h 10.12pg/ml, sgnrha (P<0.05), 48h E 2,, 96h (P<0.05), sgnrha (P< 0.05), 192h (P<0.05) sgnrha 6h, 48h 10.15pg/ml,, 192h (P>0.05) sgnrha E 2 (P>0.05) 3,,, ;,,,,

, D., E., F. 48h (P>0.05), 192h, (P<0.05) 192h (sgnrha HCG, P<0.05), sgnrha T HCG T (P>0.

5 ( ) (Cabrita et al, 2006),,,, (Zohar et al, 2001) Vermeirssen (1998),, ( T 11-KT), 17,20βdihydroxy-4-pregnen-3-one (17,20 β-p), ph (Nagahama, 1994),, HCG,, (Billard et al, 1982), (2005), LHRH-A (Epinephelus akaara), (Mylonas et al, 2001), (Vermeirssen et al, 2000),,,,, (Mylonas et al, 1997), (Lim et al, 2004), - - (GtH, Gonadotropin Hormone), ( )(Mylonas et al, 1997) (Vermeirssen et al, 2000) GtH,, (Hippoglossus hippoglossus)(vermeirssen et al, 2000) sgnrha, 3, (spermatocrit) 3 30d Methven (1992),,,,, T (6h) 96h, HCG 96h sgnrha,,,,,, (Harmin et al, 1993) (Vermeirssen et al, 1998) (2006) (LHRH-A) (hcg), T 6h, 12h,, (2005), LHRH-A, T 11- (11-KT), (Rhombosolea tapirina), GnRHa,,,,, GtH E 2 T, (Chang et al, 1999),, 6d,, (Pankhurst et al, 2000),, (Mañanós et al, 2002),,,

et al, 2000) GtH,, (Hippoglossus hippoglossus)(vermeirssen et al, 2000) sgnrha, 3, (spermatocrit) 3 30d Methven (1992),,,,, T (6h) 96h, HCG 96h sgnrha,,,,,, (Harmin et al, 1993) (Vermeirssen et

6 6 : (Verasper moseri) 1175,,,,,,,,, , 29(6): ,, :, 80,,, LHRH-A., 29: ,,, , 28: ,,, , 6(4): Alveriño J M R, Peleteiro J B, Inducción y sincronización de la puesta en el rodaballo (Scophthalmus maximus) con LHRHa: influencia del estado de desarrollo gonadal. In: Cerviño A, Landín A, Coo A D et al ed. Proc IV th National Congress on Aquaculture. Spain: Illa de Arousa (Vilanova de Arousa), Berlinsky D L, William K, Hodson R G et al, Hormone induced spawning of summer flounder Paralichthys dentatus. J World Aqua Soc, 28: Berlinsky D L, King V W, Smith T I J et al, Induced ovulation of southern flounder Paralichthys lethostigma using gonadotropin releasing hormone analogue implants. J World Aquacult Soc, 27: Billard R, Fostier A, Weil C et al, Endocrine control of spermatogenesis in teleost fish. Can J Fish Aquat Sci, 39: Cabrita E, Soares F, Dinis M T, Characterization of Senegalese sole, Solea senegalensis, male broodstock in terms of sperm production and quality. Aquaculture, 261: Chang C F, Hung C Y, Chiang M C, The concentration of plasma sex steroids and gonadal aromatase during controlled sex differentiation in grey mullet, Mugil cephalus. Aquaculture, 177: Clearwater S J, Crim L W, Milt quality and quantity produced by yellowtail flounder (Pleuronectes ferrugineus) following GnRH-analogue treatment by microspheres or pellet. In: Goetz F W, Thomas P ed. Proc V th Symp on the Reproductive Physiology of Fish. Austin: 13 Harmin S A, Crim L W, Influence of gonadotropic hormone-releasing hormone analog (GnRH-A) on plasma sex steroid profiles and milt production in male winter flounder, Pseudopleuronectes americanus (Walbaum). Fish Physiol Biochem, 10: Larsson D G J, Mylonas C C, Zohar Y et al, Gonadotropinreleasing hormone analogue (GnRH-A) induces multiple ovulations of high-quality eggs in a cold-water, batch-spawning teleost, the yellowtail flounder (Pleuronectes ferrugineus). Can J Fish Aquat Sci, 54: Lim H K, Pankhurst N W, Fitzgibbon Q P, Effects of slow release gonadotropin releasing hormone analog on milt characteristics and plasma levels of gonadal steroids in greenback flounder, Rhombosolea tapirina. Aquaculture, 240: Mañanós E, Carrillo M, Sorbera L A et al, Luteinizing hormone and sexual steroid plasma levels after treatment of European sea bass with sustained-release delivery systems for gonadotropin-releasing hormone analogue. J Fish Biol, 60: Methven D A, Crim L W, Norberg B et al, Seasonal reproduction and plasma levels of sex steroids and vitellogenin in Atlantic halibut (Hippoglossus hippoglossus). Can J Fish Aquat Sci, 49: Mylonas C C, Scott A P, Vermeirssen E L M et al, Changes in plasma gonadotropin II and sex steroid hormones, and sperm production of striped bass after treatment with controlled-release gonadotropin-releasing hormone agonist-delivery systems. Biol Reprod, 57: Mylonas C C, Zohar Y, Use of GnRHa-delivery systems for the control reproduction in fish. Rev Fish Biol and Fisheries, 10: Nagahama Y, Endocrine regulation of gametogenesis in fish. Int J Dev Biol, 38: Pankhurst N W, Effects of gonadotropin releasing hormone analogue, human chorionic gonadotropin and gonadal steroids on milt volume in the New Zealand snapper, Pagrus auratus (Sparidae). Aquaculture, 125: Pankhurst N W, Poortenaar C W, Milt characteristics and plasma levels of gonadal steroids in greenback flounder Rhombosolea tapirina following treatment with exogenous hormones. Mar Freshw Behav Physiol, 33: Vermeirssen E L M, Scott A P, Mylonas C C et al, Gonadotrophin-releasing hormone agonist stimulates milt fluidity and plasma concentrations of 17, 20 β-dihydroxylated and 5 β-reduced, 3 α-hydroxylated C21 steroids in male plaice (Pleuronectes platessa). Gen Comp Endocrinol, 112: Vermeirssen E L M, Shields R, Mazorra de Quero C et al, Gonadotrophin-releasing hormone agonist raises plasma concentrations of progestogens and enhances milt fluidity in male Atlantic halibut (Hippoglossus hippoglossus). Fish Physiol Biochem, 22: Weil C, Crim L W, Administration of LHRH analogues in

Proc IV th National Congress on Aquaculture. Spain: Illa de Arousa (Vilanova de Arousa), 31 36 Berlinsky D L, William K, Hodson R G et al, 1997.

7 various ways: effect on the advancement of spermiation in prespawning landlocked salmon, Salmo salar. Aquaculture, 35: Zohar Y, Mylonas C, Endocrine manipulations of spawning in cultured fish: from hormones to genes. Aquaculture, 197: EFFECTS OF EXOGENOUS HORMONES INDUCTION ON THE SPERM QUALITY OF BARFIN FLOUNDER VERASPER MOSERI XU Yong-Jiang 1, 2, LIU Xue-Zhou 1, 2, WANG Yan-Yan 1, 2, LIU Xin-Fu 1, 2 (1. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fisheries Sciences, Qingdao ; 2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Qingdao, ) Abstract The effects on exogenous hormones including human chorionic gonadotropin (HCG) and salmon gonadotropin releasing hormone analogue (sgnrha) on sperm quality of male barfin flounder Verasper moseri were investigated in the present study. Results showed that exogenous hormone induction can greatly improve the sperm quality of males with milt of high viscosity, including increased milt fluidity, higher sperm motility and longer sperm life span, and these were associated with elevated serum sex steroids level. The increased expression level of sex steroid peaked at 9h post hormonal induction for both groups, which was in accordance with the sperm quality elevation. The sgnrha induction beard better effect than that of HCG induction although there was no significant difference observed between sgnrha and HCG induction. Results from this study can serve as useful tools in acquiring milt with high quality and could be beneficial for artificial insemination in barfin flounder culture. Key words Barfin flounder Verasper moseri, Exogenous hormones induction, Sperm quality

Aquaculture, 197: 99 136 EFFECTS OF EXOGENOUS HORMONES INDUCTION ON THE SPERM QUALITY OF BARFIN FLOUNDER VERASPER MOSERI XU Yong-Jiang 1, 2, LIU Xue-Zhou 1, 2, WANG Yan-Yan 1, 2, LIU Xin-Fu 1, 2 (1.

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<4D F736F F D DD6DCF6A9D2C120C6EB2020F6CE2DBACBBAEC2DC1F52E646F63> 热带海洋学报 JOURNAL OF TROPICAL OCEANOGRAPHY 2010 年第 29 卷第 6 期 :130 136 海洋生物学 http://jto.scsio.ac.cn; http://www.jto.ac.cn 齐鑫 1 1,, 黄海 2, 周雯伊 1, 尹绍武 1, 张勇 2, 刘晓春 2, 陈国华 1, 林浩然 (1., 570228; 2., 510275) 1, 2