Ahstract. 1. Introduction

Transcription

1 POLSKIE ARO-!IWUM HYDROBIOLOGll (Pol. Arch. Hydrobiol.) Dariusz Kucharczyk 1, Roman Kujawa 1, Andrzej Mam carz 1, Elzbieta W y s z o m i r s k a2 Ar tilicial spawning in hr eam (Abramis brama (L.)) 1Department of Fisheries, Olsztyn University of Agriculture & Technology, Oczapo ws kiego 5, Olsztyn-Korto wo, Poland 2Department of Chemical Analysis, Drug Institute, 30/34 Chdms ka St., Warsa w, Poland Ahstract Induced spawning in bream, Abramis branw (L.), a fe w weeks before natural spa wning was studied using acetone dried common carp pituitary with human chorionic gonadotropin (HCG ). The total dose administered for fish was 4.0 mg kg-1 carp pituitary with or without S300 IU HCG kg-1 for females and 2.0 mg kg-1 carp pituitary with IU HCG kg-1 for males. Statistically significant differences of quantity of mill in treated and control groups were found; generally males fr om control group produced relatively lo w quantity of milt. The time of ovulation was not synchronous. Shortest latency as well as spa wning success, expressed as a value of Se (spawning e fectiveness coefficient) was higher in fish treated with highest dose of hormones. Key words: Abramis brama, artificial spawning 1. Introduction Obtaining the gametes is one of the import<jnt problems in some cyprinid fishes (Ahmad Tajuddinelal. 1986;T halathiahelal. 1988;Kozl owski 1994, Ya ron 1995). Bream (Abramis bromo L.) is one of the most important freshwater species in Poland and some other Europeon countries (G I u b o k o v cl al. 1991); average annual bream catch in Poland ranged from 20 (Szcze rbowski 1985) to 50 kg ha-1 (Brylinska, Ta d a j e w s k a 19 86). Prolong<ttion of spawning season is very important in some research, such as g enet ic engineering and cryopreservation (D 'I h r ows k i cl al ). In Kortowskie Llke bre a m usually spawn in the beginning of June. In this paper we present the method of spawning stimulation in bream specimens a few weeks before natural spawning. *Proceedings of the international conference "Fish Reproduction "96", 9-12 September 19 96, C..cskc Budcj ov ice, Czech Republic.

2 202 D. Kucharczyk et al. 2. Material and methods Bream (A. brama) spawners were caught in the Kortowskie Lake (Olsztyn, Poland) in end of Marc h Fis h were transported to the hatchery and fish sex was chec ked using method described by Kujawa, Kucharczy k (1996). Males and females were kept in separate 1000 dm 3 tan ks at controlled temperature (:!: 0.5" C) and photoperiod (12 hours). During fi ve days water temperature was gr adu ally raised from 8 to 1 T C. Fis h were fed twice daily. Then the fish were treated with series of hormonal injections (Table I). Before manipulations fis h were anaesthetised wit h 2-p henoxyethanol (0.2 mg dm- 3 ). After stimulatory injection the temperature of water was raised to 20o C. Females were chec ked every half an hour between 12 and 24 hours after resolving injections and every hour between 24 and 36 hours. Eggs were stripped from the individual fe males and fe rtilized with sperm pool ed from males fr om respective groups. Table I. H ormonal treatment applied to induce artificial spawning in bream (Abramis brama L.) a few wee ks before natural spawning. In each group number of males were 6, number of females were 8. HCG - human chorionic gon adot rop in (IU kg- 1 ); CPE- carp pituitary extract (mg kg- 1 ) Date Group 1 (control)* Group 2 Group 3 Male Female Male Fem ale Male Female I UHCG loo IUHCG 100 IUHCG 200 IUHCG IUHCG 200 IU HCG 200 IUHCG 400 I UHCG IU HCG 300IU HCG 300 IUHCG 600 IU HCG IU HCG 600IU HCG 450 IUHCG 900 I UHCG IU HCG 900J UHCG 600 I UHCG 1200 I UHCG I U HCG 1200 IU HCG 750 I UHCG 1500 I UHCG IU HCG 1500 IUHCG looo IUHCG 2000 IUHCG IU HC G I800 I UHCG 1250 IU HCG 2500 IUHCG mgc PE 0.4 mgcpe 0.2 mg CPE 0.4 mgc PE mg CPE 3.6 mgcpe 1.8 mgcpe 3.6mGCPE + - injection of 0.5 cm 3 0.9% NaCI. The qua lit y of eggs was assessed a fte r fertilization as a per cen ta ge of eyed egg;. Two replicates of eggs per trial were incubated on Petri dishes at water temperature C. Spawning success was e va luat ed using the formula of Se (sp awn ing effectiveness coe ficient) according to K u c h a r c z y k cl al. ( I996h ): where: Os- ovulati ng success (No. or ovub1ting fem;dcs/no. of total females); I,-i ncubat ion success (No. of live embryos/no. of tot<d embryos) Statistical differences between groups (spermiation success and incubation s ucce ss) were ana lys ed with Duncan 's multiple range test (P < (!.05) (PI a tt!977).

3 Artificial spawning in bream Results and discussion All males from control and treated groups kept in tanks were spermiating. Males from treated groups gave more milt than that from control group (Table II). Such response is typical for cyprinids where hormonal stimulation was applied to induce spermiation (T a k ash i m a et al. 1984, K o z I o w sk i 1994). There were no statistical differences in quantity of sperm obtained from stimulated groups of males, however bream males after hormonal induction in this experiment gave much less milt ( cm 3 kg-1) than males after hormonal injection during natural spawning season (more than 3.0 cm 3 kg-1) (Kucharczyk et al. 1996a). Table II. Spawning success(:!: SD) of bream (A. brnma) induced by a series injection from HCG and C PE. Numbers of groups of fish refer to the hormonal treatment described in Table I. In column s groups with common superscript were not significantly different by Duncan's multiple range test (P < 0.05) Group Latency Survival to eyed Quantity of milt Ovulated females (cm 3 (hours after final egg stage kg-1) (%) injection) (%) 1 0.4:!: A ::?. 0.9 :!: 0.09 B 37 ::?.2.3:!: 2.91 A 30.9:!: 1.65 A :!: 0.12 B :!: A 38.1 :!: 2.20 B The highest percentage of ovulated females and highest survival to the eyed egg stage was observed in the group 3, where fish was treated with higher doses of hormones (Table II). The results obtained in this experiment was inferior as compared to artificial propagation of bream in natural spawning season, using carp pituitary extract with addition of HCG, when ovulation was observed in more than 70% of females and embryos survival to the eyed egg stage was about 70% (K u char c z y k 1996), but is similar to result obtained for bream during spawning season after induction of LH-RH with dopamine antagonists (G I u b o kov et al. 1991). The latency in bream females from this experiment was not synchronous and longer in contrast to females during natural spawning season (latency about 14 hours) (K u char c z y k 1996), although in both cases fish were treated with HCG and CPE. In bream females injected with LH-RH the latency was much longer (Giubokov et al. 1991). Ya ro n (1995) suggested that lat ency was always shorter in fish treated wilh carp pituitary extract than in those treated with other hormones. Th e longer latency in fish treated with LH-RH with dopamine antagonists, in contrast with fish treated with carp pituitary extract, may be explained because LH-RH release from the pituitary and the ovari an response to the released for hormones (sequential process), while in fish injected with carp

4 204 D. Kucharczyk et al. pituitary extract, ovarian response to the exogenous GtH was a single process (D r or i et al. 1994). Spawning effectiveness coefficient (Se), which combines the results of ovulation with incubation success, was proposed by K u c ha r c z y k et al. (1996b) to point out the most efficient hormonal treatment. In the present experiment a series of injections of higher doses of hormones (group 3) turned out to be more efficient (Se = 0.19) than that of lower doses (group 2; Se = 0.11 ). In conclusion, spawning techniques described in this paper opens a possibility to obtain viable embryos a few weeks before natural spawning season. Acknowledgments This study was supported by the Project No financed by Olsztyn University of A gr iculture & Technology, Olsztyn, Poland. 4. References Ah mad Taj u d d in, Z., Ch u a h, H.P., Ha ron, A., Za ha r i, A Induced breeding techniques for some of the cyprinid foodfishes in Malacca. Proc. Intl. Conf. Dev. Managt. Trop. Living Aquat. Resources. Serdang, Malaysia., 2-5 Aug., 1983, B r y I i n s k a, M., T ad a j e w s k a, M Leszcz (A bra m is brama L.) ( Bream (Abramis brama L.)J. In: B r y I i n s k a, M. (Ed.) Ryby slodkowodne Polski [Freshwater fish of Poland] Warszawa. Panstwowe Wydawnictwo Naukowe. D a, brows k i, K., C i ere s z k o, A., Ra ms eye r, L., Cu I v er, D., K e st em on t, P Effects of hormonal treatment on induced spermiation and ovulation in the yellow perch (Perca flavescens). Aquaculture, 120, Drori, S., Ofir, M., Levavi - Sivan, B., Yaron, Z Spawning induction in common carp (Cyprinus cg!pio) using pituitary extract or GnRH superactive analogue combined with metoclopramide: analysis of hormone profile, progress of oocyte maturation and dependence on temperature. Aquaculture, 119, Glubokov, A.I., Motloch, N.N., Sedova, M.A Effect of a synthetic LHRH analogue and dop11mine antagonists on the maturation of bream, Abramis branw L. Aquaculture, 95, K o z I o w ski, B Pr11ktyka stymula cji hormonalnej w sztucznym rozrodzie ryb karpiowatych (Practice of hormon11l stimulation of artificial spawning in cyprinid fishes]. Broszura lnstytulu Rybactwa Srodlqdowego (0/sztyn) 162, K u char c z y k, D Inzynieria genomow11 lcszcza, Ahramis brama (L.) (Genome engineering in bre11m, Abramis hrama (L.)j. Ph. D. Thesis, Olsztyn University of Agriculture & Technology, O lsztyn, Ku char czyk, D., K u j a wa, R., Lu czynski, M., Glogowski, J., Babiak, I., W ys zo m irsk<t, E. 1990a. Induced spawning in bream (Ahramis hrama L.) using bre11m and carp pituitary cxtrilct and HCG. Aquaculture Research, 00, Ku charczyk, D., Kujawa, R., Mamcarz, A., Skrzypczilk, A., Wys z o m i r s k a, E. 1996b. Induced spawning in perch (I'erca flufiatilis L.) using Cilrp pituitary extract and HCG. Aquaculture Research, 00,

5 Artificial spawning in bream 205 K u jaw a, R., K u c ha r c z y k, D Przyzyciowe pobieranie oocyt6w ryb karpiowatych i okoniowatych za pomoca, katetera (Collection of oocytes from alive cyprinids and pcrcids using catheter]. Komunikaty Rybackie, 4, PI at t, C Problemy raclwnku prawdopodobienstwa [Problems of calculus of probability and statistical mathematics]. Warszawa. Panstwowe Wydawnictwo Naukowe. Szczerbowski, J.A Lcszcz (Abramis brama L.) [Bream (Abramis brama L.)J In: Szczerbowski, J.A. [Ed.J Rybactwo jeziorowe i rzeczne [Lake and ril'er fisheries] Warszawa. Panstwowe Wydawnictwo Rolnicze i Lcsne. T ak a s h i m a, F., We i I, C., B i I I a r d, R., Cr i m, LW., F os t ier, A Stimulation of spermiation by LHRH analogue in carp. /Jull. Jpn. Soc. Sci. Fish., 50, ThaI at hi a h, S., A h mad, A.O., Z a i n i, M.S Induced spawning techniques practised at Batu Berendam, Melaka, Malaysia. Aquaculture, 74, Ya r on, Z Endocrine control of gametogenesis and spawning induction in the carp. Aquaculture, 129,