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Macroscopic aspects of embryonic development of Steindachneridion parahybae. a) oocyte extruded (just after spawning, and before fertilization and hydration)-1.10 ± 0.10 mm; b) egg hydrated soon after fertilization (time zero AF)-1.90 ± 0.60 mm; c) two blastomeres (1 h 20 min AF); d) four blastomeres (1 h 40 min AF); e) eight blastomeres (1 h 50 min AF); f) sixteen blastomeres (2 h AF); g) thirty-two blastomeres (2 h 10 min AF); h) sixty-four blastomeres (2 h 40 min AF). (*): perivitelinic space; (AP): animal pole. Bars: (a-b) 0.2 mm; (c-h) 0.3 mm.
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Steindachneridion parahybae is a freshwater catfish endemic to the Paraíba do Sul River and is classified as an endangered Neotropical species. An increasing number of conservation biologists are incorporating morphological and physiological research data to help conservation managers in rescue these endangered species. This study investigated the...
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... stage. The cleavage was typically characterized by creating a cellular region above the yolk sac, which was firstly divided in the blastodisc (the first cleavage, 1 h 20 min AF), dividing this blastodisc into two blastomeres of equal size (embryonic cells, Fig. 2c). Second and third cleavages occurred, giving rise to four blastomeres (in a 2 x 2 arrangement of cells, 1 h 40 min AF, Fig. 2d Table 2. Water quality during the period of artificial reproduction at CESP Fish Farm. Data are presented as the mean ± standard error. however some blastomeres completely covered others. Beyond the sixth cleavage, the number of small cells became increasingly higher and it was more difficult to discern any stereotypical arrangement of the blastomeres, besides; the cell cycles begin to lose synchrony and any patterns of cleavage plan. The morula stage was characterized by successive cells divisions (more than sixty-four blastomeres) and arranged in a "half-berry"-like shape; the blastula stage was identified when the periblast and blastoderm region or embryonic cells became clear, and the animal pole presented a cup-like shape (without identification of embryonic cells boundaries) up to the beginning of the cell movement; when it began to characterize the gastrula stage, i.e., in this stage it was observed the occurrence of morphogenetic movement, named epiboly, which resulted in a rearrangement of the blastoderm relative to the yolk mass, and ultimately forms the germinative follicles and establishes the embryonic axis. The morphogenetic movement was divided in three steps: epiboly of 25% (Fig. 3a), epiboly of 50% (Fig. 3b), and epiboly of 90% (Fig. 3c); and subsequently, the final gastrula stage, and blastopore closure (Fig. 3d). The morula stage occurred between 2 h 50 min and 11 h 20 min ...
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... stage. The cleavage was typically characterized by creating a cellular region above the yolk sac, which was firstly divided in the blastodisc (the first cleavage, 1 h 20 min AF), dividing this blastodisc into two blastomeres of equal size (embryonic cells, Fig. 2c). Second and third cleavages occurred, giving rise to four blastomeres (in a 2 x 2 arrangement of cells, 1 h 40 min AF, Fig. 2d Table 2. Water quality during the period of artificial reproduction at CESP Fish Farm. Data are presented as the mean ± standard error. however some blastomeres completely covered others. Beyond the sixth cleavage, the number of small cells became increasingly higher and it was more difficult to discern any stereotypical arrangement of the blastomeres, besides; the cell cycles begin to lose synchrony and any patterns of cleavage plan. The morula stage was characterized by successive cells divisions (more than sixty-four blastomeres) and arranged in a "half-berry"-like shape; the blastula stage was identified when the periblast and blastoderm region or embryonic cells became clear, and the animal pole presented a cup-like shape (without identification of embryonic cells boundaries) up to the beginning of the cell movement; when it began to characterize the gastrula stage, i.e., in this stage it was observed the occurrence of morphogenetic movement, named epiboly, which resulted in a rearrangement of the blastoderm relative to the yolk mass, and ultimately forms the germinative follicles and establishes the embryonic axis. The morphogenetic movement was divided in three steps: epiboly of 25% (Fig. 3a), epiboly of 50% (Fig. 3b), and epiboly of 90% (Fig. 3c); and subsequently, the final gastrula stage, and blastopore closure (Fig. 3d). The morula stage occurred between 2 h 50 min and 11 h 20 min ...
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... stage. Initially, before hydration, it was possible to visualize that the eggs were yellow, nonadhesive, and ovoid in shape with an average diameter of 1.10 ± 0.10 mm (Fig. 2a). After fertilization and hydration of eggs, easily evident (within 10-30 min AF), fertilization activates cytoplasmatic movements (cytoplasm streams towards the animal pole, where the nucleus is positioned, to form a prominent layer of cytoplasm known as blastodisc), and the average diameter of eggs increased to about 1.90 ± 0.60 mm (Fig. 2b), characterized by a large perivitelline space (Fig. 2b) that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). The yolk was concentrated in the vegetative pole, while the cytoplasm, nucleus and organelles were distributed over the animal pole (Fig. ...
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... stage. Initially, before hydration, it was possible to visualize that the eggs were yellow, nonadhesive, and ovoid in shape with an average diameter of 1.10 ± 0.10 mm (Fig. 2a). After fertilization and hydration of eggs, easily evident (within 10-30 min AF), fertilization activates cytoplasmatic movements (cytoplasm streams towards the animal pole, where the nucleus is positioned, to form a prominent layer of cytoplasm known as blastodisc), and the average diameter of eggs increased to about 1.90 ± 0.60 mm (Fig. 2b), characterized by a large perivitelline space (Fig. 2b) that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). The yolk was concentrated in the vegetative pole, while the cytoplasm, nucleus and organelles were distributed over the animal pole (Fig. ...
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... stage. Initially, before hydration, it was possible to visualize that the eggs were yellow, nonadhesive, and ovoid in shape with an average diameter of 1.10 ± 0.10 mm (Fig. 2a). After fertilization and hydration of eggs, easily evident (within 10-30 min AF), fertilization activates cytoplasmatic movements (cytoplasm streams towards the animal pole, where the nucleus is positioned, to form a prominent layer of cytoplasm known as blastodisc), and the average diameter of eggs increased to about 1.90 ± 0.60 mm (Fig. 2b), characterized by a large perivitelline space (Fig. 2b) that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). The yolk was concentrated in the vegetative pole, while the cytoplasm, nucleus and organelles were distributed over the animal pole (Fig. ...
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... stage. Initially, before hydration, it was possible to visualize that the eggs were yellow, nonadhesive, and ovoid in shape with an average diameter of 1.10 ± 0.10 mm (Fig. 2a). After fertilization and hydration of eggs, easily evident (within 10-30 min AF), fertilization activates cytoplasmatic movements (cytoplasm streams towards the animal pole, where the nucleus is positioned, to form a prominent layer of cytoplasm known as blastodisc), and the average diameter of eggs increased to about 1.90 ± 0.60 mm (Fig. 2b), characterized by a large perivitelline space (Fig. 2b) that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). The yolk was concentrated in the vegetative pole, while the cytoplasm, nucleus and organelles were distributed over the animal pole (Fig. ...
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... This company had supported research on the biology of S. parahybae, especially on reproductive physiology and development biology. Studies on S. parahybae aimed mainly to establish of a protocol for artificial spawning [3,4], develop embryos and larvae [5][6][7], improve broodstock metabolism [8] and the neuroendocrine system [9][10][11], and improve the quality of gametes of S. parahybae [6,[11][12][13]. ...
... Steindachneridion parahybae females born and raised at CESP fish farm were selected based on the typical morphological characteristics of sexual maturity, according to parameters previously established for this species [3][4][5]. Captivity females (Table 1) were selected via external characteristics according to the hyperemic genital pore and the swollen abdomen. A biopsy was performed in these females, cannulating the gonoduct with a fine polyethylene catheter (5 mm external diameter) attached to a plastic syringe through the urogenital papilla. ...
... After this procedure, several oocytes were collected to examine their size, appearance, and diameter homogeneity. These are important criteria to establish the spawning readiness of the female broodstock in artificial hormonal induction [5]. The homogeneity of oocyte diameter was analyzed with a stereomicroscope (Leica S9D stereomicroscope, Leica MC170HD photographic camera, and computer image capture, Leica LAS Interactive Measurements). ...
This study summarizes new data on induced spawning of Steindachneridion parahybae, focusing on the aggressive behavior of females. This study characterizes the vasotocinergic system using immunohistochemistry, highlighting the potential influence of arginine-vasotocin (AVT) on reproductive physiology. Two experimental groups were proposed: (A) control, with one female in the aquarium, and (B) experimental, with two females in the same aquarium. Dominant (D) females presented a more aggressive behavior and did not show any injury. They apparently had a length and body mass higher than injured nondominant (ND) females. The analysis identified positive AVT immunoreactive (ir) neurons exclusively within the preoptic area, including parvocellular, magnocellular, and gigantocellular subpopulations, containing fibers-ir extending into the pituitary gland. Cellular and nuclear areas were greater in D compared to ND in the magnocellular subpopulation. There were no differences between parvocellular and gigantocellular subpopulations. There was a difference on the steroid plasma profile of cortisol (more in ND than in D) and 17α,20β-dihydroxy-4-pregnen-3-one (more in D than in ND). Furthermore, control and D females presented higher optical densities for AVT-ir, gonadotropin-releasing hormone-ir, and luteinizing hormone-ir than ND. In general, there were no differences in the results of female (control group) with D females. The AVT system is highly complex, possibly counting multiple sites of action during artificial reproduction and acting directly and/or indirectly associated with behavioral and physiological changes in S. parahybae females when induced to spawning.
... A Ladder (L), egg (1), two cells (2), eight cells (3), blastula (4), gastrula with 50% epiboly (5), segmentation with 10 somites (6), hatching (7). B Ladder (L), kidney, brain, muscle, liver, adult female gonad (1-5) and kidney, brain, muscle, liver, and adult male gonad (6-10) of the mouth until the development of the liver and pancreas, which occur in the first days (0-2 dph), in carnivorous catfish of the Neotropical region (Faccioli et al. 2016;Honji et al. 2012). The external pigments in P. mangurus also developed rapidly, making it difficult to visualize the PGCs after the 2 dph. ...
Primordial germ cells (PGCs) are embryonic pluripotent cells that can differentiate into spermatogonia and oogonia, and therefore, PGCs are a genetic source for germplasm conservation through cryobanking and the generation of germline chimeras. The knowledge of PGC migration routes is essential for transplantation studies. In this work, the mRNA synthesized from the ddx4 3′UTR sequence of Pseudopimelodus mangurus, in fusion with gfp or dsred, was microinjected into zygotes of three neotropical species (P. mangurus, Astyanax altiparanae, and Prochilodus lineatus) for PGC labeling. Visualization of labeled PGCs was achieved by fluorescence microscopy during embryonic development. In addition, ddx4 and dnd1 expressions were evaluated during embryonic development, larvae, and adult tissues of P. mangurus, to validate their use as a PGC marker. As a result, the effective identification of presumptive PGCs was obtained. DsRed-positive PGC of P. mangurus was observed in the hatching stage, GFP-positive PGC of A. altiparanae in the gastrula stage, and GFP-positive PGCs from P. lineatus were identified at the segmentation stage, with representative labeling percentages of 29% and 16% in A. altiparanae and P. lineatus, respectively. The expression of ddx4 and dnd1 of P. mangurus confirmed the specificity of these genes in germ cells. These results point to the functionality of the P. mangurus ddx4 3′UTR sequence as a PGC marker, demonstrating that PGC labeling was more efficient in A. altiparanae and P. lineatus. The procedures used to identify PGCs in P. mangurus consolidate the first step for generating germinal chimeras as a conservation action of P. mangurus.
... Nonetheless, we still depend on capturing wild broodstock and their transfer to fish farm facilities for domestication and future artificial induced spawning (AIS) in captivity. On the contrary, this S. parahybae domestication in captivity is not completely successful because female broodstock exhibit some reproductive dysfunction, as described by early biology studies with S. parahybae in captivity (Honji et al., 2012(Honji et al., , 2017(Honji et al., , 2019, and there are no studies that characterize the ovarian dynamics so far. ...
... One hundred S. parahybae adult females born and raised at the CESP fish farm c. 5 years (period they did not spawn) were selected based on the typical morphological characteristics of sexual maturity according to the previously established principles for this species (Honji et al., 2012). The specimens were randomly divided into two ponds (200 m 2 ) to characterize the annual RC. ...
... The specimens were randomly divided into two ponds (200 m 2 ) to characterize the annual RC. During this period, carnivorous fish were fed at 08.00 and 16.00 hours with commercial extruded food containing 40% crude protein at a rate of 2% biomass per day, provided twice a day and under natural photoperiod, according to previous studies (Honji et al., 2012(Honji et al., , 2017(Honji et al., , 2019. ...
To characterize the female reproductive biology of the endangered species Steindachneridion parahybae in captivity, the authors used the concentration of gonadal steroids and the oocyte development during the annual reproductive cycle (RC) and after artificial induced spawning (AIS) until 48 h. Three stages of gonadal maturation were identified, based on morphological and physiological features: early maturation or previtellogenic (PRV) oocyte, advanced maturation or vitellogenic (VTG) oocyte and regression (REG) or follicular atresia. They identified and characterized the following stages of germ cells: oogonia, perinucleolar and cortical alveoli, and VTG and atretic oocytes during RC. The oestradiol levels were higher in PRV than those in VTG and REG during the RC, whereas androgens showed higher levels of oestradiol in VTG than those in PRV and REG. The progestogen levels remained unchanged during the whole RC. The final oocyte maturation (FOM) was achieved after AIS and postovulatory follicles (POF) were identified. Plasma concentration of progestogens (17α,20β‐dihydroxy‐pregnen‐3‐one and 17α‐hydroxyprogesterone) increased considerably after AIS, remaining high up to 6 h after AIS, and progressively decreased over time after AIS. During RC, the lack of FOM and POFs reveals that captivity negatively impacts S. parahybae reproduction. Nonetheless, the VTG stage of oocytes, reached during RC, is suitable for ovulation induction with artificial hormone manipulation, enabling the reproduction of this species in captivity and being essential for the success of fish farming and/or fish conservation programmes (conservationist aquaculture).
... Moments after the eggs are fertilized and the zygote formed, the perivitelline space expands resulting in a clear separation of yolk and the outer protective chorion (Hill and Johnston, 1997;Hassan et al., 2018). This is important for the survival and healthy development of the eggs, as the chorion prevents mechanical damage and infection by microbiota (Korzelecka-Orkisz et al., 2010;Honji et al., 2012). Also, the perivitelline space, which is occupied by protective fluid, ensures the stability of the developing embryo as it cushions the embryo from external injury while permitting the exchange of dissolved gases and essential molecules to and from the yolk (Okomoda et al., 2017(Okomoda et al., , 2018a. ...
... This cleavage pattern is synchronous, regular and the blastomeres produced are well arranged on the vegetal pole (Olaniyi and Omitogun, 2014b). As the mitotic division proceeds, it leads to the formation of tinnier irregular cells that are asynchronous and heaped on the animal pole forming a 'mulberry' or 'half-berry' or 'ball-like' shape, which is impossible to count (Honji et al., 2012;Olaniyi and Omitogun, 2012). This period of division extends to cover the morula, blastula and gastrulation stages (Kimmel et al., 1995). ...
... This period is characterized by the formation of somite blocks that differentiate the different parts of the larvae from the head region (i.e. cephalic region with its polster and auditive parts) to the tail region (i.e. the caudal parts with Kupffer's vesicle) (Kimmel et al., 1995;Buzollo et al., 2011;Honji et al., 2012). ...
The study sought to investigate the chronology of events and timing of embryogenesis, as well as breeding performances of three strains of Heterobranchus longifilis from Nigeria. Fish samples were collected from Benue River in Makurdi, Niger River in Onitsha, and Rima River in Sokoto for this study. Induced spawning of the strains was carried out so that egg development could be tracked from fertilization to hatching using a simple microscope. The microphotographs obtained showed that the embryogenesis of the strains followed a similar pattern to those of other members of the family Clariidae, however with changes occurring in the specific timing of the sequences of events (i.e. interstrain and interspecies differences). When the different strains were compared, the study noted similarities ( P > 0.05) in the overall breeding performance (except for fertilization rate), survival at different stages of development, timing of embryogenesis, and larvae characteristics. The outcomes of this study, therefore, provide baseline information on what genetic improvement of the species through strain crossing can be attempted in future studies.
... Furthermore, conservation efforts in the field could be more successful if corroborated by captivity studies. Therefore, one of the first steps in helping conservation biologists rescue endangered species is the study of their development in captivity (Honji et al. 2012). Control of artificial reproduction is the basic knowledge required to improve the artificial propagation of any cultured species. ...
... After yolk sac resorption, larvae were daily fed at satiety on zooplankton issued from mono-specific culture of rotifers but also according to Honji et al. (2012) with Artemia nauplii (EG grade, INVE, Dendermonde, Belgium). Feed was supplied once hourly. ...
... Zaki and Abdula (1983) recorded short incubation time in C. gariepinus at high temperature. In the current study, fertilization rates varied from 60.30 to 68.02 % (64.87 ± 2.28 %) and were similar to those (45 to 76 %) recorded in the Asian Schilbeidae M. bleekeri (Long et al. 2008) and other siluriforms such as Steindachneridion parahybae Steindachner, 1877 (69.50 ± 10.20 %, Honji et al. 2012) and T. galeatus (60 %, Santos et al. 2013). Concerning hatching rates, they varied from 56.11 to 60.96 % (58.54 ± 1.94 %) and are lower than 85 to 90 % recorded in M. bleekeri (Long et al. 2008) and 68.33 ± 3.06 % recorded in H. bidorsalis (Olaniyi and Omitogun 2014). ...
The silver catfish Schilbe intermedius (Rüppell, 1832) is single-spawning species spread in Africa and may reach 50 cm in length. Since there are no studies on the artificial reproduction of S. intermedius, the aim of the present study was to induce for the first time spawning by hormonal injection and to determine the reproductive parameters for this species. Artificial induction was carried out by the injection of Ovaprim (0.5 ml kg− 1) to males and females. Different eggs measurements were carried out by micrometer. Egg stripping was carried out ten hours after hormone injection at 27.17 ± 0.22 °C (mean ± standard deviation) with mean spawning rate 58.15 ± 2.88 %. Relative fecundity was 613.25 ± 71.83 oocytes/g with mean diameters 0.94 ± 0.10 mm and 1.11 ± 0.06 mm, respectively, before and after eggs fertilization. Fecundity and hatching rates ranged, respectively, from 60.30 to 68.02 % and 56.11–60.96 %. Egg hatching occurred about 22 or 28 h after fertilization at 26.02 °C, and the larvae average length was 1.24 ± 0.14 mm. Yolk sac resorption occurred 48 h after hatching and larvae acquired their morphological structures in about 72 h after hatching. This information will be important in improving the artificial reproduction protocols of S. intermedius in controlled breeding programs.
... not always possible to obtain an adequate series from the larval to adult stages. The description of ontogenetic transformations is essential to the understanding of the functional trends and environmental preferences of different developmental stages (Honji et al. 2012) and can furnish valuable taxonomic information. ...
This study describes the main ontogenetic changes in the armored catfish Pterygoplichthys ambrosettii from newly hatched larva to adult. The relationship between the degree of development of newly hatched larvae and post-larvae and parental care is discussed. Ontogenetic series containing newly hatched larvae, post-larvae, and juveniles of P. ambrosettii were obtained from the Fish Culture Station of Universidade Estadual Paulista. Adult specimens were collected from the Preto River in the upper Paraná River basin. Morphological, osteological, morphometric, and meristic data and color pattern were analyzed. The species exhibited indirect development, with a distinct post-larval stage after the larval stage. Newly hatched larvae had a well-developed yolk sac and lacked pigmentation. The following characters underwent less change during development and can therefore be useful in taxonomical analyses: odontodes located posteriorly to the preopercle, number of plates on the lateral line, lower lip with three series of papillae, spots on the upper ray of the caudal fin, caudal fin similar to that of the adult, body ventrally covered with dermal plates and odontodes, small irregular spots on head, and typical ventral color pattern of the adult. The morphometric data revealed that most variation in body proportions occurred during less advanced developmental stages. Parental care influenced the ontogenetic development of newly hatched larvae.
... Marginal blastomeres have a predetermined different fate as they remain connected to the yolk cells by cytoplasm throughout the blastula stage. The marginal blastomeres underwent further structural changes like collapsing of blastomeres, and fusion of cytoplasm and nuclei with adjacent yolk cells resulting in the formation of the YSL (Kimmel et al., 1995;Honji et al., 2012;Olaniyi and Omitogun, 2013). Those blastomeres present in the penultimate layers are marginal in position and do not establish cytoplasmic connections, therefore are referred to as non-syncytial. ...
Garra gotyla is an indigenous coldwater fish of the cyprinid family and has wide geographical distribution in India as well as in other countries of Asia and Africa. Induced breeding in G . gotyla was carried out successfully for the first time and an attempt has been made to document developmental stages chronologically from the first minute of fertilization, through all stages of embryonic development until the fifth day post hatching. This experiment was carried out at 22–24°C water temperature at the Directorate of Coldwater Fisheries Research, Bhimtal, India. During the breeding trial, the fertilization rate was observed as 70–75% and hatching rate was 85–90%. The mature fertilized ova were measured as 0.8–1.0 mm in diameter and the perivitelline membrane became thick soon after fertilization and formation of the germ pole. The periods taken for complete developmental stages were recorded; cleavage stage 111 min (min post fertilization (pf)), blastulation stage 580 min (pf), neurulation and segmentation 1250 min (pf) and hatching was completed after 1420 min. The sac fry was measured as 3 mm in length and took almost 3 days for complete absorption of the yolk content. The major structural and differential changes observed are in head, tail, fins, alimentary canal, rudiments of each organ and appearance of melanophore pigmentation in the whole body. The 5-day-old larvae were measured as 6 mm in length with almost every organ fully differentiated. The present study will be utilized for large-scale production of fingerlings for stock enhancement in rivers, lakes and possibilities of genetic improvement and manipulation at the embryonic stage.
... After the acceptance of artificial feeding, most of the female fish were able to spawn and presented similar reproductive performance with wild specimens collected during the spawning season which were then presumably in good conditions. Moreover, the fertilization rates observed here (82%) was considerably higher than the presented for other neotropical Siluriformes species: 59% for Lophosilurus alexandri (Santos et al., 2013a), 60% for Trachelyopterus galeatus (Santos et al., 2013b), 69.5% for Steindachneridion parahybae (Honji et al., 2012) and 72.4% for Rhinelepis aspera (Perini et al., 2010), indicating that that our management provided gonadal maturation, with a similar egg weight and size and a complete oocyte vitellogenesis, which are important for reproductive success (Wallace & Selman, 1981). ...
Wild fish domestication can be considered a strategic approach to endangered species conservation, supporting studies and reducing economic and environmental costs. Three of the most important strategies in the domestication processes of fish are the adaptation of wild fish to captivity, the reproduction of the adapted fish and the production and maintenance of the young individuals. That being said, the present study is divided in three experiments: the 1st aimed to adapt wild Pseudopimelodus mangurus to captivity environment using different feeding approaches and a prophylactic strategie; the 2nd aimed to reproduce the adapted individuals from the 1st experiment; and the 3rd aimed to train the P. mangurus juveniles to accept commercial diets. The 1st and 2nd experiments were successful at the maintenance and artificial reproduction of P. mangurus kept in tanks between the reproductive seasons. The results suggest that the reproductive performance of animals kept in captivity (initial relative fertility-IRF = 609.25 ± 36.6 eggs/g) was similar (p > 0,05) to the performance found in wild individuals (IRF = 679.21 ± 45.66 eggs/g). Feed training of P. mangurus juveniles (3rd experiment) was also conducted, evaluating three feeding treatments with different concentrations of bovine heart and ration. At the end of the experiment, the treatment containing half bovine heart and half commercial feeding resulted in the highest values of weight gain (0.10 ± 0.16 g), specific growth rate (0.37 ± 0.11 mm), length (47.78 ± 2.35 mm) and growth (2.15 ± 2.27 mm), suggesting reasonable acceptability to artificial diets in the cultivation of this species. As conclusion, the present study contributes with the development of techniques for the domestication of fresh water fish species with commercial value or andangered of extinction, showing the domestication and reproduction of wild P. mangurus in captivity. However, more studies have to be conducted in order to improve the acceptance of artificial feeding by juveniles and to increase their survival rate.
... This failure is probably related to either water volume at the moment of fertilization, oocyte quality, or synchronization of the micropyle opening time Embryonic development of S. hilarii 5 (Mylonas et al., 2010). Compared with G2, fertilized eggs were observed in animals from G1, nevertheless compared with the other Characidae species, fertilization rates were considered lower (65%) (Luz et al., 2001;Andrade-Talmelli et al., 2002;Narahara et al., 2002;Weingartner and Zaniboni-Filho, 2005;Honji et al., 2012). Additionally, S. hilarii ATU in both groups were close to those of previous studies performed with Characidae, with only a few slight variations observed according to species. ...
... Gastrula phase is initially characterized by the first cellular movements in an event called epiboly, and is finished by closure of the blastopore, resulting in blastoderm rearrangement in relation to yolk mass Buzollo et al., 2011;Nakagui et al., 2006;Mello et al., 2018). Gastrula stage in S. hilarii was initiated at 3 h 56 min to 9 h 1 min, this time was considered late and no longer compared with other species such as P. maculatus (2 h 15 min to 5 h 0 min at 29ºC; Buzollo et al., Honji et al., 2012). Initially, organogenesis is marked by notochord development, which will set the caudal cephalic corporeal axis, allowing distinction between the embryonic axis and the yolk sac (Nakatani et al., 2001). ...
... The main features during this phase were optic vesicle maturation, a marked increase in somites, and pigmentation in distinct body regions, which occurred at 9 h 1 min to 20 h 45 min AF. At the end of organogenesis, muscular segmentation was observed, and the tail was completely released from the yolk sac (Andrade-Talmelli et al., 2001;Faustino et al., 2010a,b;Chalde et al., 2011;Honji et al., 2012). At approximately 20 h 46 min AF, an intense pigmentation in S. hilarii was observed in the cephalic and ventral posterior regions, this verified an intense muscular activity that consequently resulted in chorion rupture, initiating the hatching process. ...
The present study aimed to evaluate two different methods of artificial reproduction induction, and characterize the embryonic development of Salminus hilarii . Different than observed for other tropical fish species, artificial reproduction induction followed by hand-stripping of gametes was considered unfeasible for S . hilarii , as no gamete fertilization was observed. However, females that were induced and spawned naturally presented a fertilization rate of 65.64 ± 0.54%. With a mean temperature of 26.20 ± 0.90ºC it was possible to clearly distinguish a large perivitelline space at 14 min after fertilization (AF) and at 49 min AF more than 50% of the embryos presented two blastomeres, and these cleavages occurred until 1 h 54 min AF. The gastrula phase was characterized at 3 h 56 min AF, and blastopore closure was observed at 8 h 31 min AF. At 9 h 1 min, organogenesis started, with a clear distinction of the yolk mass, embryonic axis, cephalic and caudal regions; at 11 h 51 min AF the embryos already had advanced segmentation and a free tail. Total hatching occurred at 21 h 17 min and after opening the mouth, which occurred at 33 h 9 min, the larvae of S . hilarii presented a strong and characteristic cannibalism. This information can be considered fundamental to improving S . hilarii production in captivity and for collaboration with a conservation programme in the upper Tietê river basin.
... Durante el proceso de incubación de los huevos, la fase de hidratación es crítica, ya incrementa la flotabilidad de los huevos y la viabilidad del embrión; y luego de esta fase los huevos son más resistente a la manipulación y manejo. (Honji et al., 2012). La fuerte hidratación del huevo es una característica de los huevos de los peces reofílicos, por su condición de huevos semi-pelágicos (huevos de flotabilidad neutra) que requieren de un medio lótico para viabilizar el embrión. ...
Con el objetivo de reproducir vizcaína Curimata mivartii, ejemplares de dos años de edad (n= 54), mantenidos en estanques en tierra (0,1 pez/m2) fueron inducidos con extracto pituitario de carpa (EPC) (Argent, USA) a razón de 6 mg/Kg de peso para las hembras en dos aplicaciones (10 % y 90% con 12 horas entre aplicación); mientras que los machos fueron inducidos con 4,8 mg/Kg EPC en dosis única. Antes de la inducción hormonal a cada hembra le fue realizada biopsia para verificar su estado de maduración gonadal. La muestra de ovocitos fue tratada con solución Serra (Chemí, Colombia) y después de tres minutos, con la ayuda de un estereoscopio de luz, (Leica, EZ4 W, Alemania) en objetivo de 4x, se determinó posición de la vesícula germinal (céntrico, migrando, periférico, sin núcleo). El análisis de la biopsia ovárica mostró que las hembras se encontraban en maduración final. Los machos inducidos se encontraron en fase de espermiación; los cuales a ligera presión abdominal liberaron líquido seminal. Se estimó el índice de ovulación, fecundidad (absoluta y relativa), diámetro de los ovocitos y volumen seminal. El porcentaje de fertilidad fue estimado a las cuatro horas post-eclosión (hpe) y el porcentaje de eclosión fue estimado a las 10 hpe. Las hembras ovularon entre 5 y 6 horas (28±1,0°C) e inmediatamente los huevos fueron obtenidos por extrusión, así como el semen. La fecundidad absoluta fue de 178331,6±28773,7 ovocitos/hembra y la relativa 137,7±19,6 g ovocitos/Kg de hembra. El diámetro de los ovocitos recién desovados fue de 687,2±42,8 μm. La fertilidad fue de 90,7±1,4% y la eclosión de 85,3±1,5%. Los resultados permiten concluir que EPC a dosificación de 6mg/Kg de peso es un buen inductor de la reproducción de vizcaína, con alto índice de ovulación y altos porcentajes de fertilidad y eclosión.
