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Spawning Induction of Pejerrey Odontesthes Bonariensis in Captivity Using Sustained-Release Gonadotropin Releasing Hormone Agonist Implants

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Spawning Induction of Pejerrey Odontesthes Bonariensis in Captivity Using Sustained-Release Gonadotropin Releasing Hormone Agonist Implants Aquaculture Research, 2009, 41, 129^134 doi:10.1111/j.1365-2109.2009.02314.x Spawning induction of pejerrey Odontesthes bonariensis in captivity using sustained-release gonadotropin releasing hormone agonist implants LeandroA. Miranda & Gustavo M. Somoza Laboratorio de Ictio¢siolog|¤a yAcuicultura, Instituto de Investigaciones Biotecnolo¤gicas-Instituto Tecnolo¤gico de Chascomu¤s (IIB-INTECH) (CONICET-UNSAM), Buenos Aires, Argentina Correspondence: L A Miranda, Laboratorio de Ictio¢siolog|¤a y Acuicultura, Instituto de Investigaciones Biotecnolo¤gicas-Instituto Tec- nolo¤gico de Chascomu¤s (IIB-INTECH) (CONICET-UNSAM), Camino de Circunvalacio¤n Laguna Km, 6 (B7130IWA) Chascomu¤s, Buenos Aires, Argentina. E-mail: [email protected] Abstract nos Aires, Argentina. Its typical habitats are ‘lagu- nas’, the most common lentic water bodies of the The aim of this study was to induce and synchronize Argentinean pampas, which frequently have salinity spawning of pejerrey Odontesthes bonariensis (Valen- levels from 1 to 24 g L À 1 (Toresani, Lo¤pez & Go¤mez ciennes, 1835), using gonadotropin releasing hor- 1994). It has been reported recently that pejerrey mone agonist (GnRHa) implants. In the ¢rst has better survival and growth performance in experiment, the ovarian condition was assessed by brackish water, and the positive e¡ects of salinity on ovarian biopsies and the measurement of the genital ionic and osmoregulatory balance and mitigation of pore width (GPW). Females having the leading clutch stress response were also demonstrated (Somoza, of oocytes with a diameter of around 800^900 mm Miranda, Berasain, Colautti, Remes-Lenicov & and a GPW between 4.5 and 5.5 mm were treated Strˇssmann 2008). with GnRHa implants. Eighty per cent of females Because of the economic movement generated by spawned between 2 and 9 days after treatment, 12 pejerreyas a game ¢sh and also because of the quality days earlier than 20% of the ¢sh in the control group of its £esh, this species has been introduced in other that presented signs of spawning activity. In order to provinces of Argentina as well as in other countries avoid any possible ovarian injury and/or stress by the (Somoza et al. 2008). For this reason, it has been con- catheterization procedure, in a second experiment, fe- sidered that pejerrey aquaculture can be an alterna- males were selected only by visual inspection of the tive for ¢sh production in the Argentinean pampas, abdomen and GPW measurement. As in experiment where more than 2 million square hectometre of 1,80%of femalesspawnedbetween 2 and8 daysafter water bodies have been described (Reartes 1995; Lo¤- treatment, 8 days earlier than 30% of the ¢sh that pez, Baigu¤n, Iwaskiw, Del¢no & Pad|¤n 2001). In spite spawned in the control group. In both experiments, of this, and although some experimental trials have fertilization and hatching success were similar be- been performed, pejerrey culture has not been fully tween control and GnRHa-treated groups. These re- developed in Argentina (Miranda, Berasain, Velasco, sults clearly demonstrated that GnRHa implantation Shirojo & Somoza 2006; Somoza, Miranda, Guilgur & can advance and synchronize ovulation and spawn- Strobl-Mazzulla 2006; Somoza et al. 2008). ing in pejerrey without a¡ecting egg quality. The pejerrey is a multiple spawnner with a marked seasonal reproductive cycle under natural condi- Keywords: GnRHa implants, induced spawning, tions, with a major spawning period during spring pejerrey, reproduction and a minor one in autumn, with water temperatures between 18 and 20 1C (Calvo & Morriconi 1972; Introduction Strˇssmann 1989). The spawning events and the The pejerrey Odontesthes bonariensis is an inland length of the intervals between successive spawn- water atherinopsid ¢sh native to the Province of Bue- ings di¡ered considerably between and within r 2009 The Authors Journal Compilation r 2009 Blackwell Publishing Ltd 129 Induced spawning of pejerrey by GnRHa implants L A Miranda & G M Somoza Aquaculture Research, 2009, 41, 129^134 females. However, the number of eggs of the ¢rst In pejerrey,it has already been reported that it was spawning of a single female was estimated to be possible to induce spermiation by environmental and o4000 eggs in experimental ¢sh with a standard hormonal treatments without a¡ecting sperm con- length of 25^26cm (Strˇssmann 1989). Pejerrey’s centration and motility (Miranda, Escaray, Bustin- eggs are spherical, demersal and they have a dia- gorry & Somoza 2001; Miranda, CassaraŁ , & Somoza meter of 1.6 mm approximately.They also have a ser- 2005). These treatments included the increase of the ies of ¢laments that allow them to attach to di¡erent light phase duration, hCG, heterologous pituitary ex- substrates (GonzaŁ lez Regalado & Mastrarrigo1954). tracts and GnRHa injection. However, no attempts It has also been reported that the pejerrey repro- have been reported to induce spawning in pejerrey ductive period can be extended in captivity by ma- females. nipulating the photoperiod and temperature The aim of the present study was to optimize pejer- (Strˇssmann1989; Miranda et al. 2006). For example, rey reproduction in captivity using sustained-release it was possible to obtain reproductive activity almost GnRHa delivery systems to induce and synchronize throughout the year if the water temperature was ovulation and spawning, and thus provide basic in- maintained around17 1C (Toda,Tonami,Yasuda & Su- formation to establish a spawning protocol to obtain zuki 1995; Miranda et al. 2006). However, due to the eggs and larvae in a massive form. marked asynchrony and the relatively low fecundity of this species, a large number of mature ¢sh would be necessary to obtain enough eggs for the operation Materials and methods of a hatchery (Somoza et al. 2008). Experiment 1 Di¡erent hormones have been tested in many ¢sh species to induce ovulation and spawning in captiv- Sixty captive-reared adult pejerrey with external ity, including pituitary extracts, piscine gonadotro- signs of active reproductive status (spermiating pins and human chorionic gonadotropin (hCG). males and females with a prominent abdominal re- Furthermore, superactive agonists of gonadotropin gion) were selected from a broodstock kept at the out- releasing hormones (GnRHa) have been used suc- doors aquatic facilities of Instituto de Investigaciones cessfully to induce spawning in many species Biotecnolo¤gicas-Instituto Tecnolo¤gico de Chascomu¤s, (Mylonas & Zohar 2001) including, for example Chascomu¤s, Argentina. Females were anaesthetized di¡erent salmonids (Mylonas, Hinshaw & Sullivan with benzocaine, and the following measurements 1992), Sparus auratus (Linnaeus, 1758, Zohar 1989), were taken: standard length, total weight and the Pseudopleuronectes americanus (Walbaum, 1792, Har- genital pore width (GPW,Table 1 and Fig. 1) using a min & Crim 1992), Morone saxatilis (Walbaum, 1792, digital calibre (0.1mm). The ovarian biopsies were s Mylonas,Tabata, Langer, & Zohar1995) and Limanda performed using a 2 mm internal diameter Silastic ferruginea (Storer, 1839, Larsson, Mylonas, Zohar, & catheter (Dow Corning, Midland, MI, USA) intro- Crim 1997). However, due to the relatively short duced into the genital pore. For each female, the dia- half-life of GnRHa, some ¢sh require multiple GnRHa meters of at least 10 of the largest oocytes were injections for an e¡ective treatment (Mylonas, Sigela- measured using a stereoscope. Only those females ki, Divanach, Manìanos, Carillo & Afonso-Polyviou with oocytes with diameters between 800 and 2003). For this reason, the use of sustained-release, 900 mm were selected for this experiment (Fig.1).This GnRHa delivery systems (implants, microspheres, criterion was used by Strˇssmann (1989) because it osmotic pumps, etc.) to stimulate long-term release represents oocytes at the end of vitellogenesis/early of luteinizing hormone, which is the gonadotropin ¢nal oocyte maturation (FOM). Ten selected females responsible for maturation, has been proven to were intraperitoneally implanted with commercial be e¡ective in inducing ovulation and spawning in pellets containing 75 mg of sGnRHa (salmon type di¡erent species (Mylonas & Zohar 2001; Mylonas, GnRHa, OvaplantTM, Syndel, Vancouver, Canada). Papandroulakis, Smboukis, & Divanach 2004; These females were placed together with 10 un- Agulleiro, Anguis, Canìavate, Mart|¤nez-Rodr|¤guez, treated males in a 3000-L indoor tank (implanted Mylonas & CerdaŁ 2006; Corriero, Medina, Mylonas, group). Another10 females were subjected to a sham Abascal, De£orio, Arago¤n, Bridges, Santamaria, operation and placed in a similar tank with 10 un- Heinisch, Vassallo-Agius, Belmonte, Fauvel, Garcia, treated males (control group). The ¢sh were kept un- Gordin & De Metrio 2007; Ibarra-Castro & Duncan der simulated natural light conditions (14 h light:10h 2007). dark) with water temperature maintained at r 2009 The Authors 130 Journal Compilation r 2009 Blackwell Publishing Ltd, Aquaculture Research, 41, 129^134 Aquaculture Research, 2009, 41, 129^134 Induced spawning of pejerrey by GnRHa implants L A Miranda & G M Somoza Table 1 Pre-experimental data (means Æ SE) of pejerrey before treatment with GnRHa implants N SL (cm) W (g) GPW (mm) OD (lm) I1 10 36.1 Æ 1 463.4 Æ 37.5 5.1 Æ 0.06 880 Æ 8 C1 10 35.8 Æ 0.7 448.7 Æ 21.7 4.9 Æ 0.09 890 Æ 11 I2 10 35.4 Æ 0.2 477.1 Æ 6.3 4.9 Æ 009 NM C2 10 36.8 Æ 0.8 468.9 Æ 15.9 4.8 Æ 008 NM SL, standard length; W, weight; GPW, genital pore width; OD, oo- cyte diameter; N, number of females; NM, not measured. I1 and C1 (implanted and control females in experiment 1), I2 and C2 (implanted and control females in experiment 2). Figure 2 Photograph of late vitellogenic oocytes from pejerrey Odontesthes bonariensis collected by ovarian biopsy.Scale bar 510 0 mm. Spawning and egg quality Every morning after implantation and for 20 days, each tank was checked for the presence of spawned eggs. They were collected using nets located at the out£ow of the tanks.
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