International Journal of Biology Vol. 3, No. 1; January 2011

Transcription

1 Interntionl Journl of Biology Vol. 3, No. 1; Jnury 2011 Broken Eggs Influene on Fertiliztion Cpity n Viility of Eggs, Turiity n ph of Ovrin Flui n Fertiliztion Wter in the Enngere Cspin Brown Trout, Slmo Trutt Cspius Elmir Nghi Trizi (Corresponing uthor) Deprtment of Fishery Siene, Fulty of Nturl Resoures, Islmi Az University of Lhijn PO. Box.1616, Lhijn, Irn; memer of Young Reserhers Clu, Irn E-mil: Elmir_nghi@yhoo.om Hossein Khr Deprtment of Fishery Siene, Fulty of Nturl Resoures Islmi Az University of Lhijn, Lhijn, Irn Shn Ali Nezmi Irnin Fisheries Reserh Orgniztion, Tehrn, Irn Rez Lorestni Deprtment of Fisheries Sienes, Fulty of Nturl Resoures n Mrine Sienes Trit Mores University, Noor, Irn Somyeh Shmspour Deprtment of Fishery Siene, Fulty of Nturl Resoures Islmi Az University of Lhijn, Lhijn, Irn Astrt To ientify simple tool for quik evlution of qulity of enngere Cspin rown trout, Slmo trutt spius eggs, the hnges of ph (oth in ovrin flui n fertiliztion wter), turiity (oth in ovrin flui n fertiliztion wter), fertiliztion n eyeing rtes were investigte in ovrin flui smples ontining perfet eggs s well s ifferent onentrtions of roken eggs. The ph of ovrin flui n wter s well s fertiliztion n eyeing rtes erese signifintly (P<0.05) with inresing of roken eggs in ovrin flui. In ontrst, the turiity of ovrin flui n wter s well s mortlity rte of eggs inrese signifintly (P<0.05) with inresing of roken eggs in ovrin flui. Also, signifint orreltions (P<0.01) were foun etween mesure prmeters s follow: ph of ovrin flui vs. ph of wter n fertiliztion n eyeing rtes; turiity of ovrin flui vs. turiity of wter; turiity of ovrin flui n wter vs. fertiliztion n eyeing rtes. Our results onlue tht ph n turiity of ovrin flui n wter effetively influene on effiieny of rtifiil propgtion. Therefore, these oul e use s two simple tools for quik evlution of qulity of Cspin rown trout eggs uring rtifiil reproution in the hthery. Keywors: Broken eggs, ph, Turiity, Cspin rown trout 1. Introution The Cspin rown trout, Slmo trutt spius, is vulnerle nromous fish onsiere to iologil onservtion progrm in southern prt of Cspin Se (Kii et l., 1999; Niksirt & Aoli, 2009). During lst ees, the rtifiil see proution hs een goo strtegy for replenishing of Cspin rown trout stoks in se. As well s, numerous stuies hve een onute on the vrious spets of reproution in orer to elevtion of the effiieny of rtifiil fertiliztion. For exmple, the ryopreservtion of semen (Srvi et l., Pulishe y Cnin Center of Siene n Eution 161

2 Interntionl Journl of Biology Vol. 3, No. 1; Jnury ), sesonl spets of semen qulity (Hjirezee et l., 2010,,) n in vitro storge of ov (Niksirt et l., 2007). After ovultion (i.e. egg relese from the folliles), the eggs re plunge in ovrin flui, liqui mei ompose of orgni n inorgni omponents whih preserve eggs till spwning or stripping (Byunov et l., 2003). At the time of fertiliztion for slmonie, the tehniins of propgtion prefer to elimintion of ovrin flui from eggs sine some smples of ovrin flui inhiit the tivity of spermtozo. It is likely tht serete omponents of mge eggs (mostly yolk) n elining of ph estlish suh effet (Wilox et l., 1984; Billr, 1992), lthough the mehnism of this event is not fully unerstoo. In this stuy, we exmine the effets of roken eggs into ovrin flui on fertiliztion rte n viility of perfet eggs of Cspin rown trout. As well s, the turiity n ph of ovrin flui were mesure s two initors of egg isintegrtion in fish. Suh stuy n umulte more informtion on fertiliztion suess n oul e useful for promotion of rtifiil propgtion out this enngere fish. 2. Mterils n Methos The experiment ws rrie out t the Klrsht Slmonis Reproution Center (KSRC), Irn. The rooers were pture from spwning regions uring spwning seson (Novemer to Deemer). In KSRC, the rooers were heke up iweekly in orer to etetion of mture fish. A numer of Five n four mture femles (femles: TL= 28.2 ± 1.3 m, Weight = ± 51.4g) n mles (mles: TL= 30 ± 4.3 m, Weight = ± g) were selete respetively n trnsferre to hthery for olletion of genitl mterils. The ovrin flui ontining eggs s well s semen smples were ollete y hnstripping i.e. mssge from the nterior portion of the omen towrs the genitl ppill. Afterwr, the eggs were seprte from ovrin flui y net with tiny meshes. Then, ovrin flui smples were mixe n istriute in 6 Petri ishes ( quntity of 20 ml ovrin flui for eh Petri ish in 3 repetition) ontining 0, 1, 3, 5, 10, 20 eggs roken y slpel le respetively n 100 perfet eggs. After mixing, the ph n turiity (in ft, the Totl Suspene Soli (TSS) ws mesure s goo initor of turiity y portle set (Euteh, PCD 650)) were mesure in two stges s follow: ) ph n turiity of ovrin flui fter mixing with roken eggs. ) ph n turiity of e wter to ovrin flui ontining perfet n roken eggs fter fertiliztion with spermtozo. The ovrin flui n wter ph were etermine with n eletroe using SM102 ph Meter. For fertiliztion, 1 ml of poole semen of four mles with goo qulity (with ssessment of sperm ensity n perentge n urtion of motility oring to Hjirezee et l., 2010) ws mixe with ovrin flui ontining perfet n roken eggs for eh Petri ish n then, the eggs of eh Petri ish ws inute in seprte inutor. After pproximtely 8 ys, the fertiliztion rte ws etermine oring to Bromge n Cumrntung (1998) where the 50 eggs were smple rnomly from eh inutor n then fixe in formlin solution (5 ml formlin (40%) + 45 ml wter). then, eggs were investigte uner Stereomirosope (Meiji EMZ-1). The eggs with visile nervous or in the k of lrvl oy were onsiere s fertilize eggs n others without suh stte were onsiere s unfertilize eggs. During inution perio, the eyeing n hthing rtes etermine y n ojetive metho i.e. with ounting of eyeing n non-eyeing eggs s well s hthe n non-hthe iniviuls. 2.1 Sttistil nlysis The SPSS softwre ws use for t nlysis. The vlues of ph n turiity were norml oring to Kolmogorov Smirnov test ut euse of perentge t (fertiliztion n eyeing rte) i not hve norml istriution, proportionl t were onverte y ngulr trnsformtion (rsin p). One-wy nlysis of vrine (ANOVA) ws employe to ompre the mens of ph n turiity for ovrin flui n wter. Where signifint F-rtios were lulte y ANOVA, the Tukey test ws pplie to ientify whih mens were ifferent. Also, ll orreltions were teste using the ivrite orreltion oeffiients of Person. 3. Results The ph of ovrin flui n wter (Figure 1, P<0.05) s well s fertiliztion, eyeing n hthing rtes (Figure 2, P<0.05) signifintly erese with inresing of roken eggs in ovrin flui. In this regr, the perentge of fertiliztion n eyeing together n hthing strongly erese to zero for Petri ishes with more thn five n three roken eggs respetively. In ontrst, the turiity of ovrin flui n wter (Figure3, P<0.05) signifintly inrese with inresing of roken eggs in ovrin flui. Signifint orreltions were foun etween mesure prmeters in this stuy. These orreltions were s follow: ) positive orreltions: ph of ovrin flui vs. ph of wter n fertiliztion n eyeing rtes (Tle1, P<0.01); turiity of ovrin flui vs. turiity of wter (Tle1, P<0.01); ) negtive orreltions: turiity of ovrin flui n wter vs. fertiliztion n eyeing rtes (Tle1, P<0.01). 162 ISSN E-ISSN X

3 Interntionl Journl of Biology Vol. 3, No. 1; Jnury Disussion Severl stuies hve een fouse on etetion of initors of egg qulity in slmonie sine this prolem is n importnt onern for fish ulturists in terms of eonomy n effiient mngement of fry proution. The qulity of fish eggs n e ffete y numerous ftors, inluing genetis, nutrition, stress, helth sttus, wter temperture, hnling n time fter ovultion (for review, see Kjørsvik et l., 1990; Bromge, 1995; Izquiero et l., 2001; Shrek et l., 2001). Suh multiftoril onstitution of egg qulity mkes it iffiult to ontrol. Therefore, the ess to simple tool for evlution of egg qulity woul e useful in terms of time n ost sving. Until now, numerous of egg hrteristis hve een use for evlution of egg qulity. These hrteristis re: morphologil stte of eggs (Bromge, 1995), estimtion of the perentge of overripe eggs (Brnes et l., 2000), etermintion of the reltive weight inrese uring 30 min of hrening fter fertiliztion (Lhnsteiner & Ptzner, 2002) n mesurement of iohemil prmeters of ovrin flui n eggs (Lhnsteiner et l., 1999; Lhnsteiner, 2000). All mentione proeures re time n ost onsuming n require itionl skille lorn n equipment. Thus, the estlishment of simple n quik metho is neessry for etermintion of egg qulity of fish. As quik tests, two previous stuies hve use from ovrin flui ph (Dietrih et l., 2007) n wter turiity rete y roken eggs fter fertiliztion (Wojtzk et l., 2004) for evlution of qulity of rinow trout eggs. In this stuy, we exmine these lst methos for enngere Cspin rown trout. The suitility of these tests for this enngere speies n prevent from exeeing wste of eggs rete y mixing of poor n high qulity eggs. In Cspin rown trout, the ph of ovrin flui erese with inresing of roken eggs. As well s, the turiity of ovrin flui inrese with inresing of roken eggs. These results re in greement with previous reports on rinow trout, Onorhynhus mykiss (Wojtzk et l., 2004; Dietrih et l., 2007). The ition of roken eggs n susequent eresing of ovrin flui ph in this stuy oul e iret eviene for this lim tht ontent of eggs uses erese of ph of the fish ovrin flui. Suh elining in ph my e ue to the slightly i ph of egg ontent s well s reltively wek uffering pity of the ovrin flui. Therefore, we n onlue tht when low ph of rinow trout ovrin flui is oserve it is likely tht suh smples re ontminte with ontent of the roken eggs. In slmonis, it ws emonstrte tht the ovrin fluis with low ph n not tivte the motility of spermtozo t the time of fertiliztion (Wojtzk et l., in press) n thus, the fertiliztion rte ereses rmtilly. Suh event oul e possile for Cspin rown trout sine there ws positive orreltion etween ph of ovrin flui n fertiliztion rte. In the present stuy, the turiity of wter n ovrin flui inrese with inresing of the numer of roken eggs. Suh turiity oul e ue to the relesing of egg ontent (mostly yolk mterils) into the ovrin flui. On the other hn, negtive reltionship ws reore etween fertiliztion rte n turiity (oth in ovrin flui n wter). It is well known tht egg ontent originting from rupture egg ogultes upon ontt with wter n uses signifint reution of fertiliztion rte (Crl, 1941; Wilox et l., 1984). It is likely tht prtiles originting from roken eggs reue the fertiliztion pity through immoiliztion or gglutintion of spermtozo or ostrution of the miropyle. One wy for minimizing of these effets is using of fertiliztion solutions where these solutions prevent from ogultion of prtiles on eggs. In this stuy, it ws oserve tht with inresing of ovrin flui turiity n eresing of ovrin flui ph, the mortlity rte inrese uring inution of eggs. Therefore, in ition to inution onitions, it is likely tht the erese of ovrin flui ph in ompny with inresing of ovrin flui turiity inopportunely erese the viility of eggs uring inution. 5. Conlusion The results of present stuy showe tht ph n turiity of ovrin flui n fertiliztion wter signifintly influene on effiieny of rtifiil propgtion. Therefore, these oul e use s two simple tools for quik evlution of egg qulity in Cspin rown trout. As well s, using of fertiliztion solution is vise for Cspin rown trout sine slt solutions reue the verse effets of sttere ontent of roken eggs into ovrin flui. Aknowlegments The uthors express their sinere ppreition to the people who gve their time, vie, n support to this stuy, inluing: the mnger (Mr Rezvni) n stff of the Klrsht Slmonis Reproution Center (KSRC), Irn, for proviing fish n tehnil helps. Also, thnks re ue to Mr. M. Akri for helpful omments uring the experiment. Pulishe y Cnin Center of Siene n Eution 163

4 Interntionl Journl of Biology Vol. 3, No. 1; Jnury 2011 Referenes Byunov, L.V., Brnnikov, I.A., Dyuin, V.P., Gruslov, A.B., Semenkov, T.B., & Trenkler, I.V. (2003). Sex sterois onentrtions in Russin sturgeon (Aipenser guel ensteti Br.) serum n oelomi flui t finl ooyte mturtion. Fish Physiology n Biohemistry, 28, Billr, R. (1992). Reproution in rinow trout: sex ifferentition, ynmis of gmetogenesis, iology n preservtion of gmetes. Aquulture, 100, Brnes, M.E., Hnten, R.P., Syler, W.A., Cores, R.J. (2000). Viility of inln fll hinook slmon spwn ontining overripe eggs n the reliility of egg viility estimtes. N. Am. J. Aqu, 62, Bromge, N. (1995). Broostok mngement n see qulity-generl onsiertions. In: Bromge, N.R., & Roerts, R.J. (Es.). Broostok Mngement n Egg n Lrvl (pp. 1 25) Oxfor: Blkwell. Bromge, N.R., & R. Cumrntung. (1988). Egg proution in the rinow trout, In: Muir, J.F., Roert, R.J. (Es.). Reent vnes in Aquulture, (pp ) vol 3. Crl, G.C. (1941). Bewre of the roken eggs! A possile use of hevy losses of slmon eggs. Prog. Fish-Cult, 53, Dietrih, G.J., Wojtzk, M., S owi sk, M., Doosz, S., Ku mi ski, H., & Ciereszko, A. (2007). Broken eggs erese ph of rinow trout (Onorhynhus mykiss) ovrin flui. Aquulture, 273, Hjirezee, S., Mojzi Amiri, B., & Mirvghefi AR. (2010). Reltionships etween the Chemil Properties of Seminl Flui n the Sperm Motility Chrteristis of Cspin Brown Trout, Slmo Trutt Cspius (A Critilly Enngere Slmoni Fish). Reserh Journl of Fisheries n Hyroiology, 5, Hjirezee, S., Mojzi Amiri, B., & Mirvghefi, A.R. (2010). Chnges in Sperm Proution, Sperm Motility, n Composition of Seminl Flui in Cspin Brown Trout, Slmo trutt spius, Over the Course of Spwning Seson. Journl of Applie Aquulture, 22, Hjirezee, S., Mojzi Amiri, B., Mirvghefi, A.R., & Arshir Sheiykh Ahmi, A. (2010). Evlution of semen qulity of enngere Cspin rown trout (Slmo trutt spius) in ifferent times of spermition uring spwning seson. Czeh Journl of Animl siene, 55 (10), Izquiero, M.S., Fernnez-Plios, H., & Ton, A.G.J. (2001). Effet of roostok nutrition on reproutive performne of fish. Aquulture, 197, Kjørsvik, E., Mngor-Jensen, A., & Holmefjor, I. (1990). Egg qulity in fishes. In Blxter, J.H.S., & Southwr, A.J. (Es.). Avnes in Mrine Biology, (pp ) Lonon: Aemi Press. Kii, B.H., A. Aoli, & M. Neri. (1999). Sttus of fish fun in the south Cspin sin of Irn. Zool. Mile Est, 18, Lhnsteiner, F., Ptzner, R.A. (2002). Rinow trout egg qulity etermintion y the reltive weight inrese uring hrening: prtil stnriztion. J. Appl. Ihthyol, 18, Lhnsteiner, F., Weismn, T., & Ptzner, R.A. (1999). Physiologil n iohemil prmeters for egg qulity etermintion in lke trout, Slmo trutt lustris. Fish Physiol. Biohem, 20, Lhnsteiner, F. (2000). Morphologil, physiologil n iohemil prmeters hrterizing the overripening of rinow trout eggs. Fish Physiol Biohem, 23, Niksirt, H., Srvi, K., Mojzi Amiri, B., Krmi, M., & Htef A. (2007). In vitro storge of unfertilize ov of enngere Cspin rown trout (Slmo trutt spius) in rtifiil mei. Animl Reproution Siene, 100, Niksirt, H., & Aoli, A. (2009). The sttus of the ritilly enngere Cspin 345 rown trout, Slmo trutt spius, uring reent ees in the southern Cspin Se Bsin. Zoo. Mile Est, 46, Srvi, K., Niksirt, H., Mojzi Amiri, B., Mirtori, S.M., Rfiee, G.R., & Bkhtiyri, M. (2006). Cryopreservtion of semen from the enngere Cspin rown trout (Slmo trutt spius). Aquulture, 256, Shrek, C.B., Contrers-Snhez, W., & Fitzptrik, M.S. (2001). Effets of stress on fish reproution, gmete qulity, n progeny. Aquulture, 197, Wilox, K.W., Stoss, J., & Donlson, E.M. (1984). Broken eggs s use of infertility of oho slmon gmetes. Aquulture, 40, ISSN E-ISSN X

5 Interntionl Journl of Biology Vol. 3, No. 1; Jnury 2011 Wojtzk, M., Kowlski, R.C., Doosz, S., Goryzko, K., minski, H.K., Glogowski, J., & Ciereszko, A. (2004). Assessment of wter turiity for evlution of rinow trout (Onorhynhus mykiss) egg qulity. Aquulture, 242, Tle 1. Correltions etween mesure prmeters from Cspin rown trout (inluing: ovrin flui ph, ph of fertiliztion wter, ovrin flui turiity, turiity of fertiliztion wter, fertiliztion n eyeing rtes). Sttistilly signifint reltionships re inite s follows: ** P < 0.01, Dt: ivrite oeffiient. Wter turiity Ovrin flui turiity ph of wter ph of ovrin Fertiliztion rte (%) Eyeing rte (%) Hthing rte (%) flui Wter turiity º Ovrin flui turiity ** º ph of wter ** ** º ph of ovrin flui ** ** ** º Fertiliztion (%) rte ** ** ** ** º Eyeing rte (%) ** ** ** ** ** º 0.90 Hthing rte (%) ** ** ** ** ** ** º ph e f wter ovrin flui Numer of roken eggs Figure 1. The hnges of ph in fertiliztion wter n ovrin flui of Cspin rown trout in reltion to the numer of roken eggs. Mens with sme supersripts re not signifintly ifferent (P> 0.05) Pulishe y Cnin Center of Siene n Eution 165

6 Interntionl Journl of Biology Vol. 3, No. 1; Jnury Numer of roken eggs Fertiliztion rte (%) Eyeing rte (%) Hthing rte (%) Figure 2. The hnges of fertiliztion, eyeing n hthing rtes of Cspin rown trout eggs in reltion to the numer of roken eggs. Mens with sme supersripts re not signifintly ifferent (P> 0.05) Turiity (ppt) e e f f Wter turiity (Y xis:left) Ovrin flui turiity (Y xis: right) Numer of roken eggs Figure 3. The hnges of turiity in fertiliztion wter n ovrin flui of Cspin rown trout in reltion to the numer of roken eggs. Mens with sme supersripts re not signifintly ifferent (P> 0.05) 166 ISSN E-ISSN X